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Biomedical subjects

M W Griffiths

Publications and source records attributed to M W Griffiths.

At least 37 records · Page 2Linked to original sources
PubMed2000

Isolation and characterization of Carnobacterium, Lactococcus, and Enterococcus spp. from cooked, modified atmosphere packaged, refrigerated, poultry meat.

The microbiota of commercially produced, cooked and modified atmosphere packaged poultry meat was followed during storage at 3.5 degrees C for up to 7 weeks. The dominant microbiota consisted of Lactococcus raffinolactis (117 isolates), Carnobacterium divergens (61 isolates), Carnobacterium piscicola (11 isolates), Lactococcus garvieae (four isolates), Lactococcus lactis (one isolate) and Enterococcus faecalis (three isolates). All isolates were screened for production of bacteriocins. Only C. piscicola isolates produced an inhibitory substance active against other lactic acid bacteria and against several Listeria spp. Species-specific polymerase chain reaction (PCR) primers were used for the differentiation of Carnobacterium, L. raffinolactis, L. lactis, and L. garvieae strains associated with the modified atmosphere packaged poultry products. No false PCR products were observed with other closely related bacterial species.

Animals↗
PubMed2000

Survival of bifidobacteria in yogurt and simulated gastric juice following immobilization in gellan-xanthan beads.

A novel acid-stable bead made of gellan gum and xanthan gum was used to immobilize Bifidobacteria. The beads (0.75% gellan and 1% xanthan gum) had an average diameter of 3 mm and did not shrink in 25% lactic acid solution, pH 1.5, or 20% acetic acid solution, pH 1.5, after storage at 4 degrees C for 4 weeks. Bifidobacterium infantis ATCC 15697, the most acid-tolerant strain tested, was immobilized in gellan-xanthan beads and its survival in peptone water, pH 4, pasteurized yogurt, and simulated gastric juice was monitored. In peptone water, pH 4, the reduction in cell count of immobilized cells of B. infantis ATCC 15697 was not significantly different from that obtained with free cells during 6 weeks of storage at 4 degrees C. However, counts of immobilized cells of B. infantis ATCC 15697 remained significantly higher than free cells (P < 0.0001) when both were exposed to simulated gastric juices at pH 2.5, 2.0 and 1.5. At pH 2.5, the viable count of free cells dropped from 1.23 x 10(9) CFU/ml to an undetectable level (< 10 CFU/ml) in 30 min, while the viable count of immobilized cells decreased by only 0.67 log cycle in the same time period. Immobilized cells also survived significantly better than free cells (P <0.05) in pasteurized yogurt after refrigerated storage for 5 weeks.

Bifidobacterium↗
PubMed2000

A comparison of the bioscreen method and microscopy for the determination of lag times of individual cells of listeria monocytogenes.

Lag phase durations (tLag) of individual Listeria monocytogenes cells were analysed using the NightOwl Molecular Imaging System, and results were compared with mean individual cell lag times (tL) obtained from the detection time (td) method using Bioscreen. With Bioscreen, an average tL of 6.39+/-0.89 h was obtained from five separate experiments. With the NightOwl method, an average tLag of 2.73+/-0.06 h was obtained from three experiments consisting of eight total replicates. Lag values from the NightOwl and Bioscreen are related by the equation: tLag = tL + DT, where DT is the doubling time. The equivalent tLag mean value for the Bioscreen method was 7.11+/-0.84 h. Individual lag times measured by both methods were normally distributed (r2 for Bioscreen and NightOwl ranged from 0.951 to 0.999 and from 0.884 to 0.982, respectively). The results suggest that the NightOwl method can provide accurate estimates of individual cell lag times, which will facilitate the development of combined discrete continuous models for bacterial growth.

Biosensing Techniques↗
PubMed2000

Survival of listeria innocua in salmon following cold-smoke application.

The ability of Listeria innocua to survive on salmon fillets during cold smoking in a commercial processing plant was investigated using a central composite rotatable response surface design to examine smoking temperatures in the range of 18 to 30 degrees C and a smoke time from 2 to 14 h. Smoke temperature did not significantly (P < 0.05) reduce counts of L. innocua on the salmon. However, the smoking time had a significant effect on L. innocua. The smoking time was directly related to the reduction in count (R2 = 0.831), and a 3-log cycle reduction in count was observed when the smoking time was 12 h. The reduction in L. innocua levels on the fish was unaffected by the pH, water activity, and salt concentration of the fillet.

Animals↗
PubMed2000

Evaluation of the Reveal and SafePath rapid Escherichia coli O157 detection tests for use on bovine feces and carcasses.

The Reveal (Neogen Corp., Lansing, Mich.) and SafePath (SafePath Laboratories LLC, St. Paul, Minn.) tests were evaluated for their performance as beef fecal and beef carcass Escherichia coli O157:H7 monitoring tests. Agreement between these tests and a reference test system was determined using naturally contaminated bovine feces and beef carcasses. The reference system utilized immunomagnetic separation with plating onto cefixime, tellurite, sorbitol MacConkey agar, followed by colony testing using a serum agglutination test for the O157 antigen. Relative to this reference method, the Reveal test showed a sensitivity of 46% and a specificity of 82% on bovine feces and a specificity of 99% on carcass samples. The SafePath test, demonstrated a significantly higher sensitivity at 79% and a similar specificity of 79%. On carcass samples the SafePath test performed similarly to the Reveal test, demonstrating a specificity of 100% relative to the reference system. There was an insufficient number of E. coli O157-positive carcass samples to estimate precisely the sensitivity of these two methods. Both methods show promise as rapid carcass monitoring tests, but further field testing to estimate sensitivity is needed to complete their evaluation. The proportion of positive fecal samples for E. coli O157:H7 by the reference method ranged from 10.2% to 36% in 10 lots of cattle with an overall mean of 17.3% (39/225). Quarter carcass sponging of 125 carcasses revealed 1.6% positive for the pathogen (2/125).

Abattoirs↗
PubMed1999

Cloning and sequencing of the gene encoding universal stress protein from Escherichia coli O157:H7 isolated from Jack-in-a-Box outbreak.

A gene encoding the universal stress protein (uspA) was cloned from Escherichia coli O157:H7 isolated from a Jack-in-a-Box outbreak. The nucleotide sequence was determined and the amino acid sequence predicted. The gene was found to be highly conserved and shared 97% homology with the same gene of E. coli K-12. Within an 800-base pair (bp) region sequenced, a 15-bp mismatch existed between E. coli K-12 and O157:H7, of which 11 base pairs were internal to the structural gene of uspA. However, only an area with a 4-bp mismatch caused an amino acid substitution, changing an alanine to an arginine at position 140.

Amino Acid Sequence↗
PubMed1998

Quantitative risk assessment of human listeriosis from consumption of soft cheese made from raw milk.

Microbial hazards have been identified in soft cheese made from raw milk. Quantification of the resulting risk for public health was attempted within the frame of the Codex Alimentarius Commission, 1995 approach to quantitative risk assessment, using Monte Carlo simulation software. Quantitative data could only be found for Listeria monocytogenes. The complete process of cheese making was modeled, from milking to consumption. Using data published on the different sources of milk contamination (environment and mastitis) and bacterial growth, distributions were assumed for parameters of the model. Equations of Farber, J.M., Ross, W.H., Harwing, J. (1996) for general and at-risk populations were used to link the ingested dose of L. monocytogenes to the occurrence of listeriosis. The probability of milk contamination was estimated to be 67% with concentration ranging from 0 to 33 CFU ml-1. The percentage of cheese with a predicted concentration of L. monocytogenes greater than 100 CFU g-1 was low (1.4%). The probability of consuming a contaminated cheese serving was 65.3%. Individual annual cumulative risk of listeriosis, in a population each consuming 50 servings of 31 g, ranged from 1.97 x 10(-9) to 6.4 x 10(-8) in a low-risk sub-population and 1.04 10(-6) to 7.19 10(-5) in a high-risk sub-population. The average number of expected cases of listeriosis per year was 57 for a high-risk sub-population and one for a low-risk healthy sub-population. When the frequency of environmental milk contamination was reduced in the model and L. monocytogenes mastitis was eliminated, the expected incidence of listeriosis decreased substantially; the average number of expected cases was reduced by a factor of 5. Thus the usefulness of simulation to demonstrate the efficiency of various management options could be demonstrated, even if results should be interpreted with care (as many assumptions had to be made on data and their distributions.

Animals↗
PubMed1998

Identification of contamination sources of Bacillus cereus in pasteurized milk.

In order to determine the sources of Bacillus cereus in pasteurized milk, a total of 232 milk samples from various sampling points along milk processing lines and 122 environmental swabs were collected in two dairy plants between March and September, 1996. The incidence of B. cereus vegetative cells in raw milk from the plants was low (< or = 10%). However, the incidence and the average counts of B. cereus spores in the raw milk were very high and similar to those of B. cereus vegetative cells in pasteurized milk or final products after enrichment (> 80% and 1.1 x 10(5) cfu ml(-1), respectively). The incidence and average count of both vegetative cells and spores of B. cereus in environmental swabs was low. Using the microbial identification system (MIDI), a library of B. cereus fatty acid profiles comprising 229 B. cereus isolates from milk samples and environmental swabs was constructed using a critical Euclidian distance of 6.0 units as the cut-off value. Using this library, the relationship between 546 B. cereus isolates from the different sampling points along the milk processing lines and the environmental swabs was determined. Most B. cereus isolates obtained from the pasteurized milk and final products belonged to the same sub-groups as the B. cereus strains germinated from spores in raw milk. Furthermore, specific sub-groups were found in pasteurized milk, different dairy plants and at different sampling times. The results suggested that B. cereus spores in raw milk were the major source of B. cereus in pasteurized milk and that post-pasteurization contamination along the milk processing lines was possibly a minor source of B. cereus in pasteurized milk.

Animals↗
PubMed1998

Detection of pathogenic Yersinia enterocolitica using a digoxigenin labelled probe targeting the yst gene.

A 145 base pair digoxigenin-d-UTP-labelled probe, specific for pathogenic Yersinia enterocolitica heat-stable enterotoxin yst gene, was prepared by PCR. The probe was used in DNA-DNA colony hybridization and dot-blot hybridization assays. The specificity of the probe was confirmed using 52 strains representing all Yersinia spp., except Y. pestis. Out of a total of 25 Y. enterocolitica strains screened, the probe correctly identified all 18 pathogenic strains. Among the other Yersinia spp. screened, only one strain of Y. kristensenii was positively detected by the yst probe but could be differentiated by its weak signal response as compared with that obtained by pathogenic strains of Y. enterocolitica.

Bacterial Toxins↗
PubMed1998

PCR differentiation of Escherichia coli from other gram-negative bacteria using primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein.

A PCR-based test was developed to differentiate Escherichia coli from other Gram-negative bacteria. The assay employed primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein (uspA) and was found to be highly specific for E. coli. An 884 base pair (bp) specific product was amplified from all of the E. coli tested (n = 45), whereas no amplification was associated with non-E. coli Gram-negative bacteria (n = 11).

Bacterial Proteins↗
PubMed1998

Milk catalase activity as an indicator of thermization treatments used in the manufacture of cheddar cheese.

Pilot-scale studies were carried out to determine the effect of different heat treatments on catalase activity during the manufacture and maturation of Cheddar cheese. Three trials were conducted to monitor catalase activity using disk flotation and polarographic methods. Cheese was manufactured from raw milk and from milk that had been treated at 60, 65 and 72 degrees C for 16 s using a high temperature, short time heat exchanger. Catalase activity was also determined in samples of commercial milk and in samples of mild, medium, sharp, and extra sharp Cheddar cheeses obtained from different manufacturers in order to verify that the enzyme could be used as an indicator of the type of heat treatment applied to cheese milk. Catalase activity was present in cheese made from raw milk but was only present at low concentrations in cheese manufactured from thermized milk. However, high catalase activity was observed in commercial samples of sharp and extra sharp Cheddar cheese that was apparently due to the growth of catalase-producing yeasts in the cheese during maturation.

Animals↗
PubMed1998

Survival of bioluminescent Listeria monocytogenes and Escherichia coli O157:H7 in soft cheeses.

Pasteurized and raw milks that had been inoculated at 10(4) cfu/ml with bioluminescent strains of Listeria monocytogenes and Escherichia coli O157:H7 were used in the manufacture of Camembert and Feta cheeses with or without nisin-producing starter culture. Survival of both organisms was determined during the manufacture and storage of Camembert and Feta cheeses at 2 +/- 1 degree C for 65 and 75 d, respectively. Bacterial bioluminescence was used as an indicator to enumerate the colonies plated on selective Listeria agar and on MacConkey agar. Escherichia coli O157:H7 survived the manufacturing process of both cheeses and was present at the end of the storage period in greater numbers than in the initial inoculum. At the end of 75 d of storage, E. coli O157:H7 was found in the brine of Feta cheese. The counts of L. monocytogenes increased as the pH of the Camembert cheese increased, and there were significant differences between the counts from samples taken from the inside and the counts from samples obtained near the surface of the cheese. The Feta cheese that contained nisin was the only cheese in which L. monocytogenes was at the level of the initial inoculum after 75 d of storage.

Cheese↗
PubMed1998

Use of an autobioluminescent Salmonella hadar to monitor the effects of acid and temperature treatments on cell survival and viability on lactic acid-treated poultry carcasses.

To determine the long-term effects of a lactic acid rinse on viability and recovery of pathogens, Salmonella Hadar was isolated from poultry and bioluminescent constructs obtained by transformation with the lux (CDABE) gene cassette from Photobacterium phosphoreum. Results indicated that the transformed Salmonella Hadar lux was otherwise phenotypically similar to the wild-type strain. Viability studies were performed by measuring luminescence following lactic acid treatment of turkey breast and subsequent storage at -12, 0, 5, and 10 degrees C. The ability of the S. Hadar lux strain to recover was determined by monitoring light output after incubation at 22 degrees C for 10 h. The results showed that metabolic activity was significantly (P < 0.05) affected by lactic acid and by storage temperatures of -12, 0, and 5 degrees C. The lowest recovery rate was observed after rinsing with lactic acid and storing at 5 degrees C. The study demonstrated that bacterial bioluminescence is an effective way of monitoring in "real time" the ability of bacteria to recover from stress.

Abattoirs↗
PubMed1998

Repeatability of the Petrifilm HEC test and agreement with a hydrophobic grid membrane filtration method for the enumeration of Escherichia coli O157:H7 on beef carcasses.

The Petrifilm HEC test (3M Canada Inc., London, Ontario), a quantitative microbiological test for Escherichia coli O157:H7, was evaluated for its performance as a beef-carcass monitoring test. Test repeatability and agreement and agreement with an E. coli O157:H7 detection method using a hydrophobic grid membrane filter (HGMF) overlaid onto cefixime-tellurite-sorbitol MacConkey agar (CT-SMAC) followed by a latex agglutination test for the O157 antigen were determined by using pure cultures of E. coli O157:H7, beef samples experimentally contaminated with bovine feces containing E. coli O157:H7, and naturally contaminated beef carcasses of unknown E. coli O157:H7 status from a local abattoir. The Petrifilm HEC test showed excellent repeatability and excellent agreement with the HGMF-CT-SMAC method when test samples were obtained from pure cultures and experimentally contaminated meat. All 125 naturally contaminated beef carcasses surveyed were negative for E. coli O157:H7 with both microbial methods. The Petrifilm HEC test, however, demonstrated a significantly lower proportion of cross-reactive organisms (false-positive reactions) than the HGMF-CT-SMAC method. Given the performance of this test coupled with its ease of use and compact size, it shows considerable promise for carcass testing where abattoir laboratory facilities are limited and as a substitute for more complex laboratory testing methods used in established laboratories.

Abattoirs↗
PubMed1998

Antimicrobial drug residues in milk and meat: causes, concerns, prevalence, regulations, tests, and test performance.

This paper presents a historical review of antimicrobial use in food animals, the causes of residues in meat and milk, the types of residues found, their regulation in Canada, tests used for their detection, and test performance parameters, with an emphasis on immunoassay techniques. The development of residue detection methods began shortly after the introduction of antimicrobials to food animal production in the late 1940s. From initial technical concerns expressed by the dairy industry to the present public health and international trade implications, there has been an ongoing need for reliable, sensitive, and economical methods for the detection of antimicrobial residues in food animal products such as milk and meat. Initially there were microbial growth inhibition tests, followed by more sensitive and specific methods based on receptor binding, immunochemical, and chromatographic principle. An understanding of basic test performance parameters and their implications is essential when choosing an analytical strategy for residue testing. While each test format has its own attributes, none test will meet all the required analytical needs. Therefore the use of a tiered or integrated system employing assays designated for screening and confirmation is necessary to ensure that foods containing violative residues are not introduced into the food chain.

Animals↗
PubMed1998

Identification of Bacillus cereus by Fourier transform infrared spectroscopy (FTIR).

The objective of this study was to evaluate the potential of Fourier transform infrared spectroscopy (FTIR) for rapid identification of Bacillus cereus isolates. Ten B. cereus group isolates (comprising B. cereus, Bacillus mycoides, and Bacillus thuringiensis strains), five other Bacillus spp., and five non-Bacillus spp. were used. Two types of media, brain heart infusion (BHI) and Trypticase soy agar (TSA), were tested. The results indicated that all B. cereus group isolates produced characteristic absorbance peaks at wave numbers between 1738 and 1740 cm-1. These peaks were not affected by the growth medium. None of the other bacteria tested showed a similar peak after growth on BHI or TSA. Absorbance peaks between 1800 and 1500 cm-1 of members of the B. cereus group had different shapes and sizes, suggesting that FTIR may be useful for rapid identification of species within the B. cereus group.

Bacillus cereus↗
PubMed1998

Characteristics of the LacTek test as applied to tissue samples: assessment of performance using incurred field samples.

The Lactek test, marketed for antimicrobial residue detection in milk, was validated for the detection of antimicrobial residues in tissues. A previous study found that the LacTek test could confidently identify tissue samples spiked with antimicrobial residues. However, the test could not reliably distinguish violative from nonviolative spiked samples relative to Canadian maximum residue limits (MRLs). The objectives of this study were to assess and compare the performance of the LacTek tests for beta-lactams, tetracyclines, gentamicin, and sulfamethazine on samples containing naturally incurred residues by running the test in parallel with the standard microbial inhibition test (MIT) presently used for the routine testing of tissues at our facility and to assess the agreement with high pressure liquid chromatographic (HPLC) determinative methods. Parallel testing with the official MIT found that the Lactek tests could be confidently used for testing tissue samples containing incurred residues. Among 1,008 MIT-positive samples, the LacTek test found that 90% contained beta-lactams and/or tetracyclines. A further 7.3% of violative residues could not be identified to an antimicrobial class. In addition, 9% of samples testing negative on the MIT were found to contain an antimicrobial residue by the LacTek tests. Comparative testing with HPLC methods found that there was very good agreement between the two tests and that most violations were due to penicillin G and oxytetracycline. Although the LacTek test cannot be used to distinguish violative from nonviolative residue levels, it does offer several advantages over the present MIT. These include speed, ease of use, the ability to identify residues to a specific class, and an improved sensitivity at the MRL level for the most commonly found antimicrobials in tissue.

Anti-Bacterial Agents↗