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Characterization of Antibiotic-Resistant Foodborne Pathogens in Fresh Produce

Investigators
Kilonzo-Nthenge, Agnes
Institutions
Tennessee State University
Start date
2010
End date
2013
Objective
Many outbreaks have been traced to fresh produce, and this will continue to occur until fresh produce growers, retail store personnel, and consumers increase their knowledge and awareness of the risks and consequences of foodborne pathogens. There is data gap between research and the level of fresh produce safety awareness in growers, retail stores, and consumers.

Research data is needed to determine all major sources of foodborne pathogens in fresh produce. Educational programs on safe ways of handling and storing fresh produce and judicious use of antibiotics on farms are essential to improve fresh produce safety. Therefore the goals of this project is to ascertain the occurrence and persistence of antibiotic-resistant foodborne pathogens in fresh produce from farms and retail stores; and to deliver educational programs on safe handling of fresh produce; and judicious used of antibiotics on farms.

Therefore, the specific objectives are to: (1) isolate and characterize microorganisms in fresh produce from retail stores and farms; (2) assess the occurrence and compare profiles, patterns, and persistence of antibiotic-resistant pathogenic microorganisms during winter, spring, and summer seasons in soils, irrigation water, raw manure, fresh produce; and (3) educate farmers and consumers on hygienic ways of growing, handling and storing fresh produce, and (4) increase undergraduate and graduate student's participation in food safety research and outreach.

The products and measurable outcomes of the proposed work include: (1) Reduction or elimination of foodborne illnesses among the consumers; (2) competitive graduates with relevant skill for diverse opportunities in the food safety; (3) increased number of knowledgeable farmers, retail store personnel, and consumers on safer food handling practices and judicious use of antibiotics on farms; (4) enhanced research and education capacity in food safety and nutrition in the school of Agriculture and Consumer sciences; (5) at least two publications and three presentations will be produced from this work; and building stronger partnerships between Tennessee State University (TSU) and USDA in fresh produce safety.

The proposed project complements continuing efforts at TSU to provide outreach services and educate our stakeholders. Results from the proposed work will be used to implement interventions to reduce illnesses associated foodborne pathogens in fresh produce.

More information
Non-Technical Summary: In the last three decades, the number of outbreaks caused by foodborne pathogens that are associated with fresh produce has become a major concern in the United States. The overall goal of this project is to determine the persistence of antibiotic resistant foodborne pathogens in fresh produce from farm and retail stores; and to deliver educational programs on safe handling of fresh produce and judicious use of antibiotics. Therefore, the specific objectives are to: (1) isolate and characterize microorganisms in fresh produce from retail stores and farms; (2) assess the occurrence and compare profiles, patterns, and persistence of antibiotic-resistant pathogenic microorganisms during winter, spring, and summer seasons in soils, irrigation water, raw manure, fresh produce,; (3) educate farmers and consumers on hygienic ways of growing, handling and storing fresh produce and judicious use of antibiotics; and (4) increase undergraduate and graduate student's participation in food safety research and outreach. The overall plan of operation will involve: (1) collecting, characterizing and analyzing microbiological samples from fresh produce obtained from farms, farmers markets, and retail stores; (2) developing educational programs on safe handling of fresh produce and judicious use of antibiotics to minimize the potential for foodborne illnesses; (3) collaborating with United States Department of Agriculture (USDA) and in food safety research to address foodborne pathogens in fresh produce; and (4) training students in food safety research. The anticipated outcome and impacts of this proposed project include: (1) provision of scientific data to USDA Food Safety and Inspection Service (FSIS), Food and Drug Administration (FDA), and Centers for Disease Control (CDC). This will provide insight on profiles of foodborne pathogens in fresh produce, which is essential in implementing prevention and control measures; (2) improved hygienic fresh produce handling practices by farmers, consumers and judicious use of antibiotics on farms; (3) a proficient food safety workforce for the 21st century; and (4) a stronger partnership between Tennessee State University (TSU), USDA and FDA agencies in fresh produce safety research. The proposed project complements continuing efforts at TSU to strengthen food safety research and outreach.

Approach: (i) Identification of microorganisms in farm environment, fresh produce in retail stores and farmers markets. Samples of soil, irrigation water, composted or raw manure, and fresh vegetables in the farm and retail stores (25 g of each or 25 ml in case of water) will be homogenized in 225 ml of lactose broth (Difco, Becton Dickinson, Sparks, Md) and enriched at 35o C for 24 h. After homogenization, a loopful of broth from each sample will be streaked in MacConkey agar plates and incubated at 37o C for 24 h. API 20E test kit (bioMerieux, Hazelwood, Mo.) will be used to identify the bacteria isolates following the manufacturer's instruction. (ii) Genetic identification of patterns and profiles of pathogenic microorganisms DNA will be isolated using the Aquapure genomic DNA isolation kit (Bio-Rad Laboratories, Hercules, and CA). The PCR assays will be performed to determine the presence of foodborne isolates from fresh produce. The amplicons will be analyzed by electrophoresis in a 1.5% agarose gels stained with ethidium bromide and photographed using a gel documentation system (Bio-Rad). Plasmids will be isolated from overnight cultures of the isolates to determine if the antibiotic-resistant genes are located in chromosomes or plasmids. The plasmids will be prepared and will be subjected to PCR assay. The sequences will be analyzed with the BLAST (National Center for Biotechnology Information) (iii) Educational program Educational program will be designed to educate the farmers on safe agricultural practices such as, avoiding planting crops on land prone to flooding, animal feeding units coming down stream for irrigating farms needs to be avoided, testing of irrigation water before application process, and to avoid application of animal manure on the fields within 120 days before harvesting, and use of stainless steel cutter for harvesting which prevents any handling by staff. We will use the information learned from the survey and the laboratory research to try and improve the practices found to be deficient in farmers and retail stores. Therefore we will develop and test educational materials targeted to the at-risk fresh produce safety practices identified in farms and retail stores. In keeping with the rapidly expanding usage of the Internet, a comprehensive web site on prevalence of foodborne pathogens in farm environment and fresh produce and ; and educational materials on fresh produce safety practices and judicious use of antibiotics on farms will be developed. Those who do not have computers or who choose to receive information via another technique can read the same basic information in a prepared brochure and advertisements. Educational materials will be made available through local agencies and offices such as extension, Head Start, health clinics, displayed at point of sale such as, retail stores, and public buses, and schools. Pre- and post-surveys will be conducted to evaluate the effectiveness of the educational materials and delivery methods.

Funding Source
Nat'l. Inst. of Food and Agriculture
Project source
View this project
Project number
TENX-1002-FS
Accession number
223667
Categories
Legislation and Regulations
Bacterial Pathogens