Source: UNIV OF MASSACHUSETTS submitted to
CLOSTRIDIUM PERFRINGENS TYPE B IN U.S RETAIL FOODS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1005604
Grant No.
2015-67030-23008
Project No.
MASW-2014-05627
Proposal No.
2014-05627
Multistate No.
(N/A)
Program Code
A1801
Project Start Date
Feb 1, 2015
Project End Date
Jan 31, 2018
Grant Year
2015
Project Director
Labbe, R. G.
Recipient Organization
UNIV OF MASSACHUSETTS
(N/A)
AMHERST,MA 01003
Performing Department
Food Science
Non Technical Summary
It has been hypothesized that a certain type of bacterium, Clostridium perfringens, may be a link to the condition known as multiple sclerosis (MS)via the production of particlar toxin produced by this organsim. The organismism iswidely found in retail meats and a certain type is responsible for foodborne illness. A small subset (toxin type) of this organism mayalso possess a particular toxin typically associated with animal disease. It isthis particular, rare type which has been proposed to be associated with MS. The work proposed will provide an assessment of how widespread this particular type of C. perfringens is in meat and poultry.
Animal Health Component
0%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71234401100100%
Goals / Objectives
Determine the distribution of the bacterim Clostriium perfringens Types B and D in retail meat and poutry. These types have been proposed as having a role in the etiology of multiple sclerosis. The presence of the enterotoxin gene in isolates will also be determined as will be spore heat resistance to evaluate the ability of the organism to survive heating procedures.
Project Methods
Method to be used are those previousl published by us and others with particular reference to the laboratory procedures of the Food and Drug Administration. Modification of methods will be introduced as needed to account for the particular characteristics food products examined. These will be, in particular, meat and poultry. Procedures include the blending, isolation and toxigenic characterization of isolates. Of particular interest will be the levels of the target organism in foods examined and the percent of samples possessing the target organism as well as their levels expressed as per gram. The polymerase chain reaction will be used to assess the potential toxigenicity of isolates and toxin types, in particlual the epsilon toxin gene.

Progress 02/01/15 to 01/31/18

Outputs
Target Audience:academic professionals, research scientists, public health authorities Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project allowed for the training of undergraduate and graduate students in microbial food safety. How have the results been disseminated to communities of interest?As journal article and national meeting of food safety professionals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The goal was to determine the presence of the Clostridium perfringens epsilon toxin gene (etx) in isolates from retail meat, poultry, spices, processed and, to a lesser extent,pet foods. The procedure involved the enumeration, isolation, and confirmation of this organism using FDA standard methods of analysis. Once confirmed as C. perfringens, isolates were assessed for the presence of etx and for the presence of the enterotoxin gene (cpe) as well, using the Polymerase Chain Reaction (PCR). 431 retail samples of fresh meat and poultry, 247 spice samples, 193 processed meat samples, and 32 pet food samples were examined for the presence and levels of C. perfringens. Generic C. perfringens was present in 43 (17%) of the spice samples, 79 (18%) of the meat and poultry samples, 4% of the processed meat and 9% of pet food samples. In total, levels of viable C. perfringensranged from 3.6 to 2400/gm from all samples. In the case of the epsilon toxin gene (etx): Four of the spice and four of the meat and poultry isolates possessed the epsilon toxin gene (etx) equivalent to their presence in approximately 1-2% of C. perfringens isolates from these sources. No epsilon toxin genes were detected in C. perfringens isolated from processed and pet foods. We conclude that, like enterotoxin-positive C. perfringens (see below),the presence of epsilon toxin-positive C. perfringens is present in retail meat, poultry, and spices at verylow levels. Using a commercial epsilon toxin assay, 5 ofthe 8 epsilon toxin gene-positive isolates produced the toxin itself in culture fluids. It is not clear why the epsilon toxin gene, though present, was not expressed in 3 of the isolates. As the gene for the epsilon toxin is carried on a plasmid, the differentlevelsof epsilon toxin in culture fluids of isolates may be a reflection ofplasmid copy number. Although the role of the epsilon toxin in animal diseases has been well established, previous, though limited, surveys of its presence in isolates from retail food has found it to be absent or rare. Our results support the conclusion that it is unlikelythatepsilon-toxin producing C. perfringens would be commonly encounteredin retail foods (perhaps in 1-2% of isolates), even though the organism is readily isolated from retailprotein foods. In the case of the enterotoxin gene (cpe): cpe was detected in: none of the meat and poultry isolates, 4% of the processed and pet food isolates and 17% of the spice isolates. Past surveys have detected enterotoxin-positive C. perfringens at level of approximately 5 or less percent of C. perfringens isolates from retail foods.

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Lee, C.-A. and Labbe, R. 2018. Distribution of enterotoxin- and epsilon-positive Clostridium perfringens in U.S. retail spices. Journal of Food Protection 81:394-399.


Progress 02/01/15 to 01/31/16

Outputs
Target Audience:academic professionals, research scientists; public health authorities Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project allows for the training of undergraduates and graduate students in micrbiual food safety. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Additional retaill samples will be obtained and will include processed meats. Confirmed isolates will examined by molecular methods for the presence of the epsilon toxin gene. Positives will be examined for the abilty to produce the epsilon toxin in vitro. The presence of other toxin genes will also be determined, in particluar that for the enterotoxin.

Impacts
What was accomplished under these goals? The goal is to determine the presence of the epsilon toxin gene (epx) in retail meat and poultry of Clostridium perfringens. The procedure initially involves the isolation and confirmation of this organism. Once confirmed the isolates are assessed for the presence of epx. Thus far 250 retaill fresh meat and poultry samples have been anaylzed towards the goal of 400-500. Thus far74 C. perfringens isolates have been obtained. Of these 50 have been confirmed as C. perfringens. Confirmation of the remaining is on-going.

Publications