Food Protection Trends - November 2009 - 782

TABLE 1. Campylobacter spp. in retail ground beef (n = 142) based on PCR Identification Genus: Campylobacter spp. Species a,b C. jejuni only C. coli only C. jejuni and C. coli C. coli and C. hyointestinalis a b Positive (%) 65 (45.8) 20 (14.1) 35 (24.6) 1 (0.7) 2 (1.4) seven isolates could not be identified to the species level. zero samples tested positive for DNA of C. fetus, C. lanienae, C. concisus or C. upsaliensis. with each meat sample to the enrichment broth. For each package of fresh retail ground beef, the plastic wrap over the middle was sliced with a sterile scalpel blade. A deep core sample of 25 g (24–26 g) of raw ground beef was removed with a sterile spoon. Each ground beef sample was placed into a 55-ounce Whirl Pak bag (82007-726, VWR International, Mississauga, ON, Canada) with 1 ml of C. jejuni solution and 100 ml of enrichment broth (Bolton broth (# CM0983 Oxoid Ltd., Basingstoke, UK) and 5% horse blood mixture) and mixed thoroughly for 30 seconds (Stomacher Lab Blender 400). The homogenate was then incubated (85% N2, 10% CO2, 5% O2) for 44 hours at 42°C and then streaked onto Karmali selective agar (Oxoid, CM935 with supplement SR0167E, Nepean, ON, Canada) and microaerobically incubated for 48–72 hours. Each culture plate was then examined visually for colonies characteristic of Campylobacter spp. (based on growth, color and morphology of the colony, and color of the cell mass). Ground beef packages were not tested for campylobacters prior to inoculation. Five packages of retail ground beef were tested at each concentration (1 × 104, 1 × 103, 1 × 102, or 1 × 101 CFU/g), and the experiment was repeated on two separate days. Each incubation of test plates included both a negative control plate and a laboratory strain C. jejuni plate as positive control. These experiments were conducted to document our ability to consistently recover C. jejuni from ground beef by use of our culture protocol. Study protocol for detection of campylobacters by use of enrichment culture The enrichment culture protocol for the study retail ground beef was the same as that already described for the experimental inoculation, except without the addition of the 1 ml of fresh C. jejuni solution. Briefly, 25 g of raw ground beef was added to 100 ml of a Bolton broth and 5% horse blood mixture in a 55-ounce Whirl Pak bag and mixed thoroughly for 30 s. The homogenate was then microaerobically incubated for 44 hours at 42°C and then streaked onto Karmali selective agar and re-incubated microaerobically at 42°C for 48–72 hours. Each culture plate was then examined visually for colonies characteristic of Campylobacter spp. Each incubation included a laboratory strain C. jejuni plate as positive control. tion and application of taxon-specific PCR for campylobacters. Each subsample (1 g) was thawed and placed in a BagPage 100 filtered blending bag (EW36840-58; Canadawide Scientific Ltd., Ottawa, ON, Canada) containing 9 ml of Columbia broth (Becton, Dickinson and Company, Sparks, NV, USA), and the sample was homogenized for 120 s at high setting in a Stomacher 80 blender (Seward Ltd., West Sussex, UK). The homogenate was then removed from the bag and centrifuged at 1,750 × g for 10 minutes, the supernatant containing Campylobacter cells was collected. To concentrate Campylobacter cells, the supernatant was centrifuged at 24,050 × g for 10 minutes, and the supernatant removed and discarded. The pellet was re-suspended in 1 ml of Columbia broth, 200 µl aliquots were placed in 2 ml tubes, an internal amplification control (IAC; 10 µl containing 700 copies/µl) was added to each tube (15), and DNA was extracted using the DNAeasy Tissue Kit (Qiagen, Missassauga, Canada) according to the manufacturer’s protocol. Direct PCR was applied for Campylobacter genus, IAC, C. jejuni, C. coli, C. fetus, C. hyointestinalis, and C. lanienae (15). In addition, nested PCR to detect C. concisus and C. upsaliensis was applied (Inglis et al., unpublished). In all instances, negative and positive PCR controls were included, and arbitrarilyselected amplicons (including weak amplicons) were sequenced to ensure specificity. Samples were deemed to be negative for Campylobacter DNA only if amplification of the IAC occurred (i.e., in the absence of a Campylobacter genus amplicon). Data analysis Descriptive analyses were conducted using SPSS (version 15.0; SPSS, Chicago, US). A second commercial software package (MLwiN version 2.02; Centre for Multilevel Modeling, Institute of Education, London, UK) was utilized for the hierarchical model analysis. The hierarchical models (9) were specified with a logit link, binomial distribution, restricted iterative generalized least square and second order penalized quasi-likelihood nonlinear estimation. The outcome was whether or not a ground beef sample was positive for Campylobacter spp. DNA. Variables included “poultry cutting” (whether or not poultry was cut or Detection of campylobacters by polymerase chain reaction (PCR) At the same time as samples were taken for culture, ground beef from approximately 10% of the 1,200 packages collected (52 of 60 stores represented) were frozen for subsequent DNA extrac- 782 FOOD PROTECTION TRENDS | NOVEMBER 2009

Food Protection Trends - November 2009

Table of Contents for the Digital Edition of Food Protection Trends - November 2009

Food Protection Trends - November 2009
Contents
Sustaining Members
Vickie’s View from Your President
Commentary from the Executive Director
Prevalence and Risk Factor Investigation of Campylobacter Species in Retail Ground Beef from Alberta, Canada
Consumer Storage Period and Temperature for Peanut Butter and Their Effects on Survival of Salmonella and Escherichia coli O157:H7
General Interest Paper – History of Consumer Food Safety Education Focus on Beef: Impact on Risk of Foodborne Illness
Award Nominations
New Members
What’s Happening in Food Safety
Industry Products
Coming Events
Advertising Index
Journal of Food Protection Table of Contents
Audiovisual Library Order Form
Booklet Order Form
Membership Application
Food Protection Trends - November 2009 - Food Protection Trends - November 2009
Food Protection Trends - November 2009 - Cover2
Food Protection Trends - November 2009 - 765
Food Protection Trends - November 2009 - Contents
Food Protection Trends - November 2009 - 767
Food Protection Trends - November 2009 - 768
Food Protection Trends - November 2009 - 769
Food Protection Trends - November 2009 - 770
Food Protection Trends - November 2009 - 771
Food Protection Trends - November 2009 - 772
Food Protection Trends - November 2009 - Sustaining Members
Food Protection Trends - November 2009 - 774
Food Protection Trends - November 2009 - 775
Food Protection Trends - November 2009 - Vickie’s View from Your President
Food Protection Trends - November 2009 - 777
Food Protection Trends - November 2009 - Commentary from the Executive Director
Food Protection Trends - November 2009 - 779
Food Protection Trends - November 2009 - Prevalence and Risk Factor Investigation of Campylobacter Species in Retail Ground Beef from Alberta, Canada
Food Protection Trends - November 2009 - 781
Food Protection Trends - November 2009 - 782
Food Protection Trends - November 2009 - 783
Food Protection Trends - November 2009 - 784
Food Protection Trends - November 2009 - 785
Food Protection Trends - November 2009 - 786
Food Protection Trends - November 2009 - Consumer Storage Period and Temperature for Peanut Butter and Their Effects on Survival of Salmonella and Escherichia coli O157:H7
Food Protection Trends - November 2009 - 788
Food Protection Trends - November 2009 - 789
Food Protection Trends - November 2009 - 790
Food Protection Trends - November 2009 - 791
Food Protection Trends - November 2009 - 792
Food Protection Trends - November 2009 - General Interest Paper – History of Consumer Food Safety Education Focus on Beef: Impact on Risk of Foodborne Illness
Food Protection Trends - November 2009 - 794
Food Protection Trends - November 2009 - 795
Food Protection Trends - November 2009 - 796
Food Protection Trends - November 2009 - 797
Food Protection Trends - November 2009 - 798
Food Protection Trends - November 2009 - 799
Food Protection Trends - November 2009 - Award Nominations
Food Protection Trends - November 2009 - 801
Food Protection Trends - November 2009 - 802
Food Protection Trends - November 2009 - 803
Food Protection Trends - November 2009 - 804
Food Protection Trends - November 2009 - 805
Food Protection Trends - November 2009 - 806
Food Protection Trends - November 2009 - 807
Food Protection Trends - November 2009 - New Members
Food Protection Trends - November 2009 - 809
Food Protection Trends - November 2009 - What’s Happening in Food Safety
Food Protection Trends - November 2009 - 811
Food Protection Trends - November 2009 - 812
Food Protection Trends - November 2009 - Industry Products
Food Protection Trends - November 2009 - 814
Food Protection Trends - November 2009 - 815
Food Protection Trends - November 2009 - Coming Events
Food Protection Trends - November 2009 - Advertising Index
Food Protection Trends - November 2009 - 818
Food Protection Trends - November 2009 - 819
Food Protection Trends - November 2009 - 820
Food Protection Trends - November 2009 - Journal of Food Protection Table of Contents
Food Protection Trends - November 2009 - Audiovisual Library Order Form
Food Protection Trends - November 2009 - Booklet Order Form
Food Protection Trends - November 2009 - Membership Application
Food Protection Trends - November 2009 - Cover3
Food Protection Trends - November 2009 - Cover4
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