Location: Quality and Safety Assessment Research Unit
Project Number: 6040-42000-044-000-D
Project Type: In-House Appropriated
Start Date: Apr 1, 2016
End Date: Mar 11, 2021
Objective:
The goal of the research is to develop and validate early, rapid, sensitive and/or high-throughput methods and techniques for detecting biological and physical hazards in poultry (food) products with optical sensing methods and instruments. Thus, the nature of the research is to combine chemistry and engineering disciplines (optical, agricultural, and food) with microbiological techniques to solve food safety detection problems in poultry (food). Specific objectives are:
Objective 1: Develop high-speed imaging methods for rapid detection of pathogens in live poultry flocks, and foodborne hazards, including foreign materials, in processed poultry products.
Sub-objective 1A: Develop Salmonella surveillance system for early detection of diseased birds.
Sub-objective 1B: Develop high-speed hyperspectral imaging methods and system for foreign material detection.
Objective 2: Develop rapid methods and protocols for early detection, identification, and quantification of pathogens in poultry products (foods) using imaging spectroscopy.
Sub-objective 2A: Develop hyperspectral microscope imaging (HMI) methods and system for early detection and identification of pathogen at the cellular level.
Sub-objective 2B: Develop fluorescence in-situ hybridization (FISH) imaging methods to identify pathogenic bacteria at the cellular level.
Sub-objective 2C: Develop nanobiosensor for pathogen detection with surface enhanced Raman spectroscopy (SERS) at the cellular level.
Sub-objective 2D: Develop methods for intervention carryover for Salmonella detection.
Sub-objective 2E: Develop methods for plate detection with optimized agar media at the colony level.
Objective 3: Develop methods to detect biofilms in poultry processing facilities with optical technologies. In developing the methods assess if any biomarkers can be identified to enhance or improve the detection sensitivity or specificity.
Approach:
Ensuring poultry meat is safe to eat is of utmost importance to producers and consumers alike and rapid and early detection of foodborne pathogenic bacteria and foreign material in poultry products is needed. This research, which is divided into three objectives, primarily investigates optical sensors for rapid or improved detection of pathogenic bacteria with imaging and spectroscopic methods. Obj. 1A: an early-warning imaging surveillance system will be developed to detect bile in poultry droppings from laying hens in their cages. These higher levels of bile have been linked to birds with very high levels of Salmonella. Spectra will be collected to optimize key wavelengths and then a color-imaging system will be optimized for wireless real-time monitoring. Obj. 1B: Building on the success of a high speed hyperspectral imaging system developed within the unit, research will be expanded to detect foreign materials in various processed poultry products. Spectral libraries of normal meat features (muscle, fat, skin) and foreign material (rubber, metal, plastics, bone) will be used to develop algorithms suitable for high-speed use and then tested in real time. Obj. 2A: Hyperspectral microscope imaging (HMI) will be used to classify and quantify pathogens commonly found in poultry and other meats. The focus will be on identifying single bacteria cells from chicken rinsate by combining cell morphology and spectral profiles into an automated method for counting and classifying pathogenic bacteria. Additionally, markers will be used to enhance detection or means to separate and concentrate the bacteria will be implemented (immunomagnetic beads). Obj. 2B: Multiplex fluorescence in-situ hybridization (m-FISH) will be combined with HMI to further enhance detection with new protocols that will combine multiplexed probes and enhanced HMI detection resulting in broader, more robust methods of identification. Obj. 2C: Surface enhanced Raman spectroscopy (SERS), utilizing aptamers or antibodies and nano-enhanced surfaces, will be studied for Salmonella detection in broiler meat. Both labeled and label-free SERS will be evaluated. Obj. 2D: At FSIS’s request, research to neutralize sanitizers, frequently used to reduce pathogens while processing poultry meat, will be conducted to prevent interference of those sanitizers on bacterial analysis. Four potential neutralizing agents (quaternary ammonium, peroxyacetic acid, acidified sodium chlorite, acid solution, and dibromodimethylhydantoin sanitizers) will be screened for efficacy. Obj. 2E: Hyperspectral imaging (HI) systems will be used to classify pathogenic serovars growing in agar plates and collaborations will explore additional agar additives (both chromogenic and non-chromogenic) that will help differentiate serovars of E. coli O157:H7 and other shiga-toxin producing E. coli (STEC). Obj. 3: First in the lab, and then in processing plants, HI systems will be used and paired with spray-on markers to enhance the detection of biofilms on equipment surfaces. This research is potentially collaborative with ARS Beltsville and will help to discriminate biofilms from other organic material.
