Progress 11/09/01 to 09/30/05
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? The phenomenon of bacteria attaching to processing plant surfaces such as metals, rubber, and plastics is a formidable obstacle for sanitizing and cleaning treatments. When bacteria attach to a surface, they produce extracellular polymers that anchor the cells and provide a favorable environment for growth and further attachment of more microbes and debris. The composite is a biofilm that is resistant to cleaners and sanitizers that is extremely difficult to remove. Our goal is to reduce the risk of foodborne disease by determining how pathogens, especially Campylobacter jejuni and Listeria monocytogenes, interact within biofilms in processing environments and to use this information to develop effective intervention strategies that minimize contamination of poultry products. Methods will be developed
to grow and characterize biofilms on surfaces and to detect target pathogens within biofilms. Diverse bacterial populations containing the pathogens will be profiled for susceptibility to control in laboratory and field environments. Physical, chemical, and alternative intervention strategies will be tested for efficacy and stability to control bacterial contamination. Understanding the formation, composition, and cellular interactions of pathogens within biofilms will enable development of exciting new interventions to counteract these processes and thereby enhance plant sanitation practices for pathogen control. The reduction of microbial pathogens in food products is the most pressing food safety problem today. Research is needed to elucidate the fundamentals of growth and survival of pathogens in situ, including their resistance to sanitation practices. The pathogen reduction technologies in this research are focused primarily on post slaughter processing. The specific goals of
this project are to: 1) elucidate the formation and composition of biofilms on processing plant surfaces and 2) develop methods to prevent the formation or facilitate removal of biofilms from surfaces to allow efficacious cleaning and sanitizing. The research to be undertaken falls under National Program 108 - Food Safety and addresses goals 2.2.1.3, 2.2.1.4, and 2.2.1.5 as described in the National Program Action Plan. Specifically these are: 2.2.1.3 Trace sources (niches) of pathogens in processing ecosystems. Determine the physiological status of microorganisms within these niches, and determine any special characteristics required for survival. Determine role of quorum sensing and nutrient availability. Identify conditions where pathogen growth is restricted. 2.2.1.4 Assess the role of biofilm formation on pathogen attachment, resistance to control measures, and transmission of pathogens to consumers. Determine the relationship of pathogens and other microorganisms within
biofilms, both on foods and within processing ecosystems. Develop model systems for characterizing attachment. 2.2.1.5 Identify new chemical and physical technologies which can interfere with pathogen adhesion to surfaces. Develop model compounds to interfere with pathogens, based on mechanisms of attachment and detachment. Measure the types and viability of pathogens on food surfaces treated with candidate inhibitors or disinfectants. Providing cost-effective improvements in food plant sanitation methods will enhance the value of agricultural products for consumers as well as for the poultry industry. Agencies in need of this research include regulatory agencies, such as Food Safety and Inspection Service (FSIS), the Animal and Plant Health Inspection Service, and the Environmental Protection Agency (EPA). Standards agencies, the National Standards Foundation (NSF) and the 3-A Symbol Council (3-A), have requested information from this research for setting standards for equipment
production. Commercial interests include poultry processors, chemical and equipment manufacturers, and consumer interests such as the US Poultry and Egg Association and National Chicken Council. 2. List the milestones (indicators of progress) from your Project Plan. Year 1 (FY2002) Bacteriological methods will be developed and applied to generate experimental models for biofilms containing pathogens at 18 months. Year 2 (FY2003) Optimum treatment parameters and reduction efficiency will be determined for electrostatic space charge system (ESCS) at 24 months. Year 3 (FY2004) The experimental model will be used to evaluate the existence of target pathogens within biofilms at 36 months. Year 4 (2005) Optimum surface treatments that combine control strategies will be determined at 48 months. Year 5 (2006) Experimental model will be used to evaluate options for controlling biofilms containing target pathogens at 60 months. 3a List the milestones that were scheduled to be addressed in FY
2005. For each milestone, indicate the status: fully met, substantially met, or not met. If not met, why. 1. An experimental model was used to continue evaluation of the existence of target pathogens within biofilms. Milestone Substantially Met 2. Optimum surface treatments that combine control strategies were determined. Milestone Substantially Met 3. An experimental model was used to evaluate options for controlling biofilms containing target pathogens. Milestone Substantially Met 3b List the milestones that you expect to address over the next 3 years (FY 2006, 2007, and 2008). What do you expect to accomplish, year by year, over the next 3 years under each milestone? This project is due to terminate 5/31/06. During FY2006 the experimental biofilm model will be used to evaluate existence of target pathogens within biofilms from processing facilities. 4a What was the single most significant accomplishment this past year? Methods were developed and applied to generate models for
biofilms containing the pathogen Campylobacter jejuni. Growth of the pathogen and other biofilm bacteria was measured for multiple nutritional and atmospheric conditions. The numbers of each type of bacteria were measured at different times during their life cycles to determine how pathogens grow within biofilms on stainless steel. The pathogen, containing a green fluorescent protein, was observed by specialized microscopy methods. These methods showed the occurrence, viability, and fluorescence of C. jejuni on stainless steel with and without incorporation into biofilms with natural populations of other bacteria. Although other bacteria were not necessary for attachment of C. jejuni to stainless steel, C. jejuni can survive many days alone, but the presence of other bacteria enhanced the growth and expansion of C. jejuni over the surface. 4b List other significant accomplishments, if any. Technology to transfer a strong negative electrostatic charge to biofilms on stainless steel has
shown promise to reduce bacterial contamination on surfaces. An electrostatic space charge system was used to treat biofilms of specific bacterial pathogens attached to stainless steel. Treatment of Campylobacter jejuni, Listeria monocytogenes, Salmonella enteritidis, Staphylococcus aureus, and Escherichia coli achieved up to a 99.9 % reduction efficiency. This technology could have applications that extend into other food processing areas, medical institutions, and the home. 4c List any significant activities that support special target populations. An electrostatic space charge system (ESCS) was tested against bacterial spores which are much more resistant to killing by other methods than vegetative cells of bacteria. Treatment of bacterial spores of Bacillus stearothermophilus achieved up to a 99.8% reduction efficiency. These results with bacterial spores suggest that the ESCS is a promising alternative treatment with the potential to kill spores without the use of toxic
chemicals. This is the first report of ionization to effectively kill bacterial spores. 4d Progress report. The effects of corrosion and biofouling are costly problems on the surface of stainless steel, the most common material in poultry processing plants. A corrosive treatment model to simulate wet-processing conditions commonly used in food processing applications was employed to test the effects of surface corrosion on bacterial attachment. Electropolished samples were significantly most resistant to bacterial attachment, before and after exposure to corrosive treatment, than stainless steel that was not electropolished. Appropriate finishes on stainless steel surfaces can improve resistance to bacterial attachment and corrosion to enhance food safety during processing. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. This research has provided the national food safety program, regulatory agencies, and the processing
industry with new information on the inhibition of bacterial attachment to processing plant surfaces, as well as characteristics of the bacteria and surfaces. Research on the incidence and level of Campylobacter jejuni in the poultry processing environment was initiated under this CRIS. Action Plan component 2.2.1.2, 2.2.1.3, and 2.2.1.4. Digital aroma technology was put to novel use to detect, compare, profile, and classify the characteristics of bacterial species important to poultry processing as potential pathogens. The substrate utilization profiles of bacterial communities (biofilm) were assessed as they developed on chicken meat samples stored for varying time periods at two temperatures commonly used in a poultry processing facility. Action Plan component 2.2.1.3 and 2.2.1.4. Bacterial isolates from poultry processing were tested for susceptibility to antibiotics commonly used in therapeutic treatment of poultry. Knowledge of the antibiotic resistance patterns will aid in
selection or inhibition of these bacterial species for further study. The beneficial vs. harmful affects of therapeutic treatment of poultry with antibiotics is hotly debated. Our results showed that resistance to antibiotics used for therapeutic treatment occurs in bacteria in the processing environment. This means that resistance to the antibiotics is carried by bacteria on poultry products. Action Plan component 2.2.1.2, 2.2.1.3, and 2.2.1.4. Appropriate finishes on stainless steel improve resistance of processing surfaces to bacterial attachment, biofilm formation, and corrosion; and thereby enhance food safety during processing. Atomic force microscopy was shown to be a useful tool to predict the potential for bacteria to attach and form biofilms on surfaces. Factors that are important for resistance of stainless steel surfaces, the most common in food processing, to bacterial contamination were determined. Our new findings, related to the bacterial resistance of treated
stainless steel and other materials are being used to set standards for equipment surfaces in meat, poultry, milk, and biotech industries. Formerly, electropolishing, the most resistant finish tested, was limited to production of shiny exteriors to enhance aesthetic appeal of equipment. Most surfaces that contact products are now electropolished in the poultry processing, milk, meat, pharmaceutical, and other wet-process industries. Action Plan component 2.2.1.3, 2.2.1.4, and 2.2.1.5. The effects of corrosion and biofouling are costly problems on the surface of stainless steel, the most common material in poultry processing plants. We employed a corrosive treatment model to simulate wet-processing conditions commonly used in food processing applications and test the effects of surface corrosion on bacterial attachment. Electropolished samples were significantly most resistant to bacterial attachment, before and after exposure to corrosive treatment, than seven other finishes tested.
Appropriate finishes on stainless steel surfaces can improve resistance to bacterial attachment, biofilm formation, and corrosion and thereby enhance food safety and economy during processing. Action Plan component 2.2.1.4 and 2.2.1.5. An electrostatic space charge system (ESCS) was tested against bacterial spores which are much more resistant to killing by other methods than vegetative cells of bacteria. Treatment of bacterial spores of Bacillus stearothermophilus achieved 99.8% reduction efficiency. Treatment of the pathogens Campylobacter jejuni, Listeria monocytogenes, Salmonella enteritidis, Staphylococcus aureus, and Escherichia coli achieved up to 99.9% reduction efficiency. These results with bacterial pathogens and spores suggest that the ESCS is a promising alternative treatment that acts without the use of toxic chemicals. Action Plan component 2.2.1.5. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to
become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Initiated Trust Agreement to develop and deliver intervention strategies against Listeria monocytogenes biofilms on food equipment.
Impacts
(N/A)
Publications
- Arnold, J.W., Boothe, D.D., Suzuki, O., Bailey, G.W. 2004. Multiple imaging techniques demonstrate the manipulation of surfaces to reduce bacterial contamination and corrosion. Journal of Microscopy. 216:215-221.
- Arnold, J.W. 2004. Complementary microscopy methods define form and function of food processing surfaces. Proceedings of the American Oil Chemists' Society Meeting. p. 35.
- Sanders, S.Q., Boothe, D.D., Frank, J.F., Arnold, J.W. 2004. Campylobacter jejuni attachment within mixed population biofilms formed on stainless steel [abstract]. American Society for Microbiology Meeting. p. 21.
- Arnold, J. W. 2005. Sanitation in poultry processing. In: Mead, G. Food safety control in the poultry industry. Cambridge, UK: Woodhead Publishing. p. 360-379
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Progress 10/01/03 to 09/30/04
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? The phenomenon of bacterial attachment to processing plant surfaces such as metals, rubber, and plastics presents a formidable obstacle for sanitizing and cleaning treatments. When bacteria attach to a surface, they produce extracellular polymers that anchor the cells and provide a favorable environment for growth and attachment of more microbes and debris. The composite is a biofilm that is resistant to cleaners and sanitizers and is extremely difficult to remove. Our goal is to reduce the risk of foodborne disease by determining how pathogens, especially Campylobacter jejuni and Listeria monocytogenes, interact within biofilms in processing environments and to use this information to develop effective intervention strategies that minimize contamination of poultry products. Methods will be developed
to grow and characterize biofilms on surfaces and to detect target pathogens within biofilms. Diverse bacterial populations containing the pathogens will be profiled for susceptibility to control in laboratory and field environments. Physical, chemical, and alternative intervention strategies will be tested for efficacy and stability to control bacterial contamination. Understanding the formation, composition, and cellular interactions of pathogens within biofilms will enable us to develop exciting new interventions to counteract these processes and thereby enhance plant sanitation practices for pathogen control. The reduction of microbial pathogens in food products is the most pressing food safety problem today. Research is needed to elucidate the fundamentals of growth and survival of pathogens in situ, including their resistance to sanitation practices. The pathogen reduction technologies in this research are focused primarily on post slaughter processing. The specific goals of
this project are to: 1) elucidate the formation and composition of biofilms on processing plant surfaces and 2) develop methods to prevent the formation or facilitate removal of biofilms from surfaces to allow efficacious cleaning and sanitizing. The research to be undertaken falls under National Program 108 - Food Safety and addresses goals 2.2.1.3, 2.2.1.4, and 2.2.1.5 as described in the National Program Action Plan. Specifically these are: 2.2.1.3 Trace sources (niches) of pathogens in processing ecosystems. Determine the physiological status of microorganisms within these niches, and determine any special characteristics required for survival. Determine role of quorum sensing and nutrient availability. Identify conditions where pathogen growth is restricted. 2.2.1.4 Assess the role of biofilm formation on pathogen attachment, resistance to control measures, and transmission of pathogens to consumers. Determine the relationship of pathogens and other microorganisms within
biofilms, both on foods and within processing ecosystems. Develop model systems for characterizing attachment. 2.2.1.5 Identify new chemical and physical technologies which can interfere with pathogen adhesion to surfaces. Develop model compounds to interfere with pathogens, based on mechanisms of attachment and detachment. Measure the types and viability of pathogens on food surfaces treated with candidate inhibitors or disinfectants. Providing cost-effective improvements in food plant sanitation methods will enhance the value of agricultural products for consumers as well as for the poultry industry. Agencies in need of this research include regulatory agencies, such as Food Safety and Inspection Service (FSIS), the Animal and Plant Health Inspection Service, and the Environmental Protection Agency (EPA). Standards agencies, the National Standards Foundation (NSF) and the 3-A Symbol Council (3-A), have requested information from this research for setting standards for equipment
production. Commercial interests include poultry processors, chemical and equipment manufacturers, and consumer interests such as the US Poultry and Egg Association and National Chicken Council. 2. List the milestones (indicators of progress) from your Project Plan. Bacteriological methods will be developed and applied to generate experimental models for biofilms containing pathogens at 18 months. Experimental model will be used to evaluate the existence of target pathogens within biofilms at 36 months. Optimum treatment parameters and reduction efficiency will be determined for electrostatic space charge system (ESCS) at 24 months. Optimum surface treatments that combine control strategies will be determined at 48 months. Experimental model will be used to evaluate options for controlling biofilms containing target pathogens at 60 months. 3. Milestones: A. List the milestones (from the list in Question #2) that were scheduled to be addressed in FY 2004. How many milestones did you
fully or substantially meet in FY 2004 and indicate which ones were not fully or substantially met, briefly explain why not, and your plans to do so. The project is ahead of schedule. FY2004 milestones -1)Bacteriological methods were developed and applied to generate experimental models for biofilms containing pathogens. 2)Optimum surface treatments that combine control strategies were determined. The methods and treatments for these milestones were included in research and incorporated into peer-reviewed journal articles. Investigations continue to optimize surface treatments for newly arising problem areas. B. List the milestones (from the list in Question #2) that you expect to address over the next 3 years (FY 2005, 2006, & 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? This project began in 2001, and will be reviewed with NP108 in 2005. The Year 4 and 5 milestones are listed below with a description of the anticipated
outcomes. The entire project is scheduled to be completed during FY 2005, and a new project will be developed to undergo OSQR review, with subsequent implementation beginning FY 2006. Year 4 (FY2005) Use experimental model, which was developed in this project, of mixed species biofilm containing C. jejuni to determine intervention strategies under food processing plant conditions. Continue investigation of optimum surface treatments that combine control strategies against pathogens. Assuming that the subsequent project plan continues the research along its present course, we anticipate accomplishing in: Year 5 (FY2006) Establish baseline data of our intervention technology against L. monocytogenes. Experimental model will be used to evaluate the existence of target pathogens and to characterize the cell-cell interactions within biofilms from processing facilities. Year 6 (FY2007) Determine mechanism of action for optimization of intervention technology against multi-species biofilms
from poultry processing equipment. Use chemical screening to optimize product formulations for efficacy against C. jejuni and L. monocytogenes within biofilms. Develop product to achieve 'zero tolerance' for L. monocytogenes. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004 (one per Research (OOD) Project): Methods were developed and applied to generate models for biofilms containing the pathogen Campylobacter jejuni. Growth of the pathogen and other biofilm bacteria was measured for multiple nutritional and atmospheric conditions. The numbers of each type of bacteria were measured at different times during their life cycles to determine how pathogens grow within biofilms on stainless steel. The pathogen, containing a green fluorescent protein, was observed by specialized microscopy methods. These methods showed the occurrence, viability, and fluorescence of C. jejuni on stainless steel with and without
incorporation into biofilms with natural populations of other bacteria. Although other bacteria were not necessary for attachment of C. jejuni to stainless steel, C. jejuni can survive many days alone, but the presence of other bacteria enhanced the growth and expansion of C. jejuni over the surface. B. Other significant accomplishment(s), if any. 1) Technology to transfer a strong negative electrostatic charge to biofilms on stainless steel has shown promise to reduce bacterial contamination on surfaces. An electrostatic space charge system was used to treat biofilms of specific bacterial pathogens attached to stainless steel (J. Arnold, Poultry Processing & Meat Quality Research Unit (PPMQ); B. Mitchell, Southeast Poultry Research Lab (SEPRL)). Treatment of Campylobacter jejuni, Listeria monocytogenes, Salmonella enteritidis, Staphylococcus aureus, and Escherichia coli achieved up to a 99.9 % reduction efficiency. This technology could have applications that extend into other
food processing areas, medical institutions, and the home. 2)The use of spores Bacillus anthracis (causative agent for anthrax) as a weapon for bioterrorism has brought the attention of the world to the need for new methods to kill bacterial spores. An electrostatic space charge system was tested against bacterial spores which are much more resistant to killing by other methods than vegetative cells of bacteria (J. Arnold, PPMQ, and B. Mitchell, SEPRL). Treatment of bacterial spores of Bacillus stearothermophilus achieved up to a 99.8% reduction efficiency. These results with bacterial spores suggest that the ESCS is a promising alternative treatment with the potential to kill spores without the use of toxic chemicals. This is the first report of ionization to effectively kill bacterial spores. 3) The effects of corrosion and biofouling are costly problems on the surface of stainless steel, the most common material in poultry processing plants. We employed a corrosive treatment model
to simulate wet-processing conditions commonly used in food processing applications and test the effects of surface corrosion on bacterial attachment (J. Arnold, PPMQ, O. Suzuki, JGC Corp., and G. Bailey, EPA). Electropolished samples were significantly most resistant to bacterial attachment, before and after exposure to corrosive treatment, than stainless steel that was not electropolished. Appropriate finishes on stainless steel surfaces can improve resistance to bacterial attachment and corrosion and thereby enhance food safety and economy during processing. C. Significant activities that support special target populations. None D. Progress Report opportunity to submit additional programmatic information to your Area Office and NPS (optional for all in-house ('D') projects and the projects listed in Appendix A; mandatory for all other subordinate projects). 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. This
research has provided the national food safety program, regulatory agencies, and the processing industry with new information on the inhibition of bacterial attachment to processing plant surfaces, as well as characteristics of the bacteria and surfaces. Research on the incidence and level of Campylobacter jejuni in the poultry processing environment was initiated under this CRIS. The work resulted in the spawn of a separate CRIS, #6612-41420-007, Microbial ecology and transmission of human pathogens in the poultry processing plant. Action Plan component 2.2.1.2, 2.2.1.3, and 2.2.1.4. Digital aroma technology was put to novel use to detect, compare, profile, and classify the characteristics of bacterial species important to poultry processing as potential pathogens. The substrate utilization profiles of bacterial communities (biofilm) were assessed as they developed on chicken meat samples stored for varying time periods at two temperatures commonly used in a poultry processing
facility. Action Plan component 2.2.1.3 and 2.2.1.4. Bacterial isolates from poultry processing were tested for susceptibility to antibiotics commonly used in therapeutic treatment of poultry. Knowledge of the antibiotic resistance patterns will aid in selection or inhibition of these bacterial species for further study. The beneficial vs harmful effects of therapeutic treatment of poultry with antibiotics is hotly debated. Our results showed that resistance to antibiotics used for therapeutic treatment occurs in bacteria in the processing environment. This means that resistance to the antibiotics is carried by bacteria on poultry products. Action Plan component 2.2.1.2, 2.2.1.3, and 2.2.1.4. Appropriate finishes on stainless steel improve resistance of processing surfaces to bacterial attachment, biofilm formation, and corrosion; and thereby enhance food safety and economy during processing. Atomic force microscopy was shown to be a useful tool to predict the potential for bacteria
to attach and form biofilms on surfaces. Factors that are important for resistance of stainless steel surfaces, the most common in food processing, to bacterial contamination were determined. Our new findings, related to the bacterial resistance of treated stainless steel and other materials, are being used to set standards for equipment surfaces in meat, poultry, milk, and biotech industries. Formerly, electropolishing, the most resistant finish tested, was limited to production of shiny exteriors to enhance aesthetic appeal of equipment. Most surfaces that contact products are now electropolished in the poultry processing, milk, meat, pharmaceutical, and other wet-process industries. Action Plan component 2.2.1.3, 2.2.1.4, and 2.2.1.5. The effects of corrosion and biofouling are costly problems on the surface of stainless steel, the most common material in poultry processing plants. We employed a corrosive treatment model to simulate wet-processing conditions commonly used in food
processing applications and test the effects of surface corrosion on bacterial attachment (J. Arnold, PPMQ, and O. Suzuki, JGC Corp.). Electropolished samples were significantly most resistant to bacterial attachment, before and after exposure to corrosive treatment, than seven other finishes tested. Appropriate finishes on stainless steel surfaces can improve resistance to bacterial attachment, biofilm formation, and corrosion and thereby enhance food safety and economy during processing. Action Plan component 2.2.1.4 and 2.2.1.5. The use of spores Bacillus anthracis (causative agent for anthrax) as a weapon for bioterrorism has brought the attention of the world to the need for new methods to kill bacterial spores. An electrostatic space charge system (ESCS) was tested against bacterial spores which are much more resistant to killing by other methods than vegetative cells of bacteria (J. Arnold, PPMQ, and B. Mitchell, SEPRL). Treatment of bacterial spores of Bacillus
stearothermophilus achieved 99.8% reduction efficiency. Treatment of the pathogens Campylobacter jejuni, Listeria monocytogenes, Salmonella enteritidis, Staphylococcus aureus, and Escherichia coli achieved up to a 99.9 % reduction efficiency. These results with bacterial pathogens and spores suggest that the ESCS is a promising alternative treatment that acts without the use of toxic chemicals. Action Plan component 2.2.1.5.
Impacts
(N/A)
Publications
- Arnold, J.W., Boothe, D.D., Mitchell, B.W. 2003. Electronic space charge system, an alternative intervention strategy for biofilm reduction. Biofilms, an American Society for Microbiology Conference. Victoria, Canada. October 2003.
- Davis, L.D., Williams, A., Mitchell, B.W., Arnold, J.W. 2003. The effect of electrostatic treatment against salmonella enteritidis on plastic surfaces. American Society for Microbiology.
- Arnold, J.W., Boothe, D.D., Suzuki, O., Bailey, G.W. 2003. Multiple imaging techniques demonstrate the manipulation of surfaces to reduce bacterial contamination and corrosion. Applied Physics International Meeting. p. 292.
- Arnold, J.W., Boothe, D.D., Mitchell, B.W. 2004. Use of negative air ionization for reducing bacterial pathogens and spores on stainless steel surfaces. Journal of Applied Poultry Research. 13:200-206.
- Sanders, S.Q., Hinton Jr, A., Frank, J.F., Arnold, J.W. 2004. Evaluation of fatty acid versus substrate utilization for microbial identification of bacterial isolates associated with fresh poultry. [abstract] Minorities in Agriculture, Natural Resources, and Related Sciences Conference Proceedings. Meeting Abstract.
- Arnold, J.W., Suzuki, O. 2003. Effects of corrosive treatment on stainless steel surface finishes and bacterial attachment. Transactions of the American Society of Agricultral Engineers. 46(6):1595-1602.
- Arnold, J.W., Boothe, D.D. 2004. Low levels of bacteria found on rubber picker fingers during processing. [abstract] Poultry Science. 82(suppl.1).
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Progress 10/01/02 to 09/30/03
Outputs
1. What major problem or issue is being resolved and how are you resolving it? The phenomenon of bacterial attachment to processing plant surfaces such as metals, rubber, and plastics presents a formidable obstacle for sanitizing and cleaning treatments. When bacteria attach to a surface, they produce extracellular polymers that anchor the cells and provide a favorable environment for growth and attachment of more microbes and debris. The composite is a biofilm that is resistant to cleaners and sanitizers and is extremely difficult to remove. Our goal is to reduce the risk of foodborne disease by determining how pathogens, especially Campylobacter jejuni and Listeria monocytogenes, interact within biofilms in processing environments and to use this information to develop effective intervention strategies that minimize contamination of poultry products. The specific objectives of this project are to: 1) elucidate the formation and composition of biofilms on processing
plant surfaces and 2) develop methods to prevent the formation or facilitate removal of biofilms from surfaces to allow efficacious cleaning and sanitizing. Methods will be developed to grow and characterize biofilms on surfaces and to detect target pathogens within biofilms. Diverse bacterial populations containing the pathogens will be profiled for susceptibility to control in laboratory and field environments. Physical, chemical, and alternative intervention strategies will be tested for efficacy and stability to control bacterial contamination. Understanding the formation, composition, and cellular interactions of pathogens within biofilms will enable us to develop exciting new interventions to counteract these processes and thereby enhance plant sanitation practices for pathogen control. 2. How serious is the problem? Why does it matter? The reduction of microbial pathogens in food products is the most pressing food safety problem today. Research is needed to elucidate the
fundamentals of growth and survival of pathogens in situ, including their resistance to sanitation practices. The pathogen reduction technologies in this research are focused primarily on post slaughter processing. Providing cost-effective improvements in plant sanitation methods will enhance the value of agricultural products for consumers as well as for the poultry industry. Agencies in need of this research include regulatory agencies, such as FSIS, the Animal and Plant Health Inspection Service, and the Environmental Protection Agency (EPA). Standards agencies, ANS, NSF, and the 3-A Symbol Council (3-A), have requested information from this research for setting standards for equipment production. Commercial interests include poultry processors, chemical and equipment manufacturers, and consumer interests such as the US Poultry and Egg Association and National Chicken Council. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been
assigned? This project relates to the ARS National Program Action Plan((108) Food Safety and Animal Products. The research seeks to improve programs for the American public(s health, safety, and wellbeing by providing knowledge to determine the fundamental parameters of microbial interactions with processing surfaces. Specifically, the project falls into categories 2.1.1.6, 2.2.1.5, and 2.2.1.6 of the National Program (108) Action Plan. 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2003: Testing of technology to transfer a strong negative electrostatic charge to biofilms on stainless steel has shown promise to reduce bacterial contamination on surfaces. An electrostatic space charge system was used to treat biofilms of specific bacterial pathogens attached to stainless steel (J. Arnold, Poultry Processing Meat Quality Research Unit (PPMQ); B. Mitchell, Southeast Poultry Research Lab (SEPRL)). Treatment of
Campylobacter jejuni, Listeria monocytogenes, Salmonella enteritidis, Staphylococcus aureus, and Escherichia coli achieved up to a 99.9 % reduction efficiency. This technology could have applications that extend into other food processing areas, medical institutions, and the home. B. Other Significant Accomplishment(s): 1) The use of spores Bacillus anthracis (causative agent for anthrax) as a weapon for bioterrorism has brought the attention of the world to the need for new methods to kill bacterial spores. An electrostatic space charge system was tested against bacterial spores which are much more resistant to killing by other methods than vegetative cells of bacteria (J. Arnold, PPMQ, and B. Mitchell, SEPRL). Treatment of bacterial spores of Bacillus stearothermophilus achieved up to a 3 log reduction with a 99. 8% reduction efficiency. These results with bacterial spores suggest that the ESCS is a promising alternative treatment with the potential to kill spores without the use of
toxic chemicals. 2) The effects of corrosion and biofouling are costly problems on the surface of stainless steel, the most common material in poultry processing plants. We employed a corrosive treatment model to simulate wet-processing conditions commonly used in food processing applications and test the effects of surface corrosion on bacterial attachment (J. Arnold, PPMQ, and O. Suzuki, JGC Corp.). Electropolished samples were significantly most resistant to bacterial attachment, before and after exposure to corrosive treatment, than seven other finishes tested. Appropriate finishes on stainless steel surfaces can improve resistance to bacterial attachment and corrosion and thereby enhance food safety and economy during processing. 3) We need to know how the pathogen Campylobacter jejuni interacts with other bacteria from processing environments so that we can develop effective intervention strategies that minimize contamination of poultry products. Methodology was developed for
culturing C. jejuni within mixed populations of biofilms (J. Arnold, PPMQ, D. Boothe, PPMQ, S. Sanders, PPMQ, J. Frank, University of Georgia). Growth was assessed with six media, two temperatures, and two atmospheric conditions to develop culture methods for liquid media that could be used for a biofilm model, and growth kinetics were followed at four cell densities to determine temporal compatibility with biofilm mixtures. Data can now be collected to quantitatively define and monitor progress of biofilm communities containing C. jejuni. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Methods were developed to measure attached bacteria and biofilm formation on surfaces, surface materials that were resistant to bacterial attachment were identified, and physical and electrochemical treatments of stainless steel were assessed for inhibition of bacterial attachment and biofilms. Atomic force microscopy was shown to be a
useful tool to predict the potential for bacteria to attach and form biofilms on surfaces. Factors that are important for resistance of stainless steel surfaces, the most common in food processing, to bacterial contamination were determined. The efficacy of new on-line technology for reduction of fecal contamination on broiler carcasses was tested. Digital aroma technology was put to novel use to detect, compare, profile, and classify the characteristics of bacterial species important to poultry processing as potential pathogens. The substrate utilization profiles of bacterial communities (biofilm) were assessed as they developed on chicken meat samples stored for varying time periods at two temperatures commonly used in a poultry processing facility. 6. What do you expect to accomplish, year by year, over the next 3 years? This project will have completed a five year cycle in FY 2005, so planning is only for the next 2 FY's. FY2004-Bacteriological methods will be developed and
applied to generate experimental models for biofilms containing pathogens. Optimum surface treatments that combine control strategies will be determined. FY2005-Experimental model will be used to evaluate the existence of target pathogens within biofilms from processing facilities. 8. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: This does not replace your peer-reviewed publications listed below). Arnold, J.W. Invited presentation, "Electropolishing and surface finish" to American Society for Mechanical Engineers Surface Finish Committee for Bioprocessing Equipment (an American National Standard). San Diego, CA. October 31, 2002.
Impacts
(N/A)
Publications
- Arnold, J.W. Advances in food sanitation: Use of intervention strategies. Schmidt, R.H., Rodrick, G.E., editors. John Wiley Sons, New York. Food Safety Handbook. 2003. p. 337-351.
- Arnold, J.W., Mitchell, B.W. An electrostatic space charge system to control bacteria and spores on surfaces. American Society for Microbiology. 2002. Abstract p. A3.
- Boothe, D.H., Arnold, J.W. Susceptibility of bacterial isolates from poultry products to therapeutic veterinary antibiotics. Journal of Food Protection. 2003. v. 66. p. 94-102.
- BOOTHE, D.D., ARNOLD, J.W. NUTRIENT SUBSTRATES USED BY BACTERIAL ISOLATES FROM THE POULTRY PROCESSING ENVIRONMENT. JOURNAL OF POULTRY SCIENCE. 2002.
- Yates, I.E., Arnold, J.W., Basinger, W., Bacon, C.W. Fusarium verticillioides induction of maize seed rot and its control. Canadian Journal of Botany. 2003. v. 81. p. 422-428.
- Yates, I.E., Arnold, J.W., Basinger, W., Bacon, C.W. Food additive controls growth of Fusarium verticillioides. International Congress of Plant Pathologists. 2003. p. 451.
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Progress 10/01/01 to 09/30/02
Outputs
1. What major problem or issue is being resolved and how are you resolving it? The phenomenon of bacterial attachment to processing plant surfaces such as metals, rubber, and plastics presents a formidable obstacle for sanitizing and cleaning treatments. When bacteria attach to a surface, they produce extracellular polymers that anchor the cells and provide a favorable environment for growth and attachment of more microbes and debris. The composite is a biofilm that is resistant to cleaners and sanitizers and is extremely difficult to remove. Our goal is to reduce the risk of foodborne disease by determining how pathogens, especially Campylobacter jejuni and Listeria monocytogenes, interact within biofilms in processing environments and to use this information to develop effective intervention strategies that minimize contamination of poultry products. The specific objectives of this project are to: 1) elucidate the formation and composition of biofilms on processing
plant surfaces and 2) develop methods to prevent the formation or facilitate removal of biofilms from surfaces to allow efficacious cleaning and sanitizing. Methods will be developed to grow and characterize biofilms on surfaces and to detect target pathogens within biofilms. Diverse bacterial populations containing the pathogens will be profiled for susceptibility to control in laboratory and field environments. Physical, chemical, and alternative intervention strategies will be tested for efficacy and stability to control bacterial contamination. Understanding the formation, composition, and cellular interactions of pathogens within biofilms will enable us to develop exciting new interventions to counteract these processes and thereby enhance plant sanitation practices for pathogen control. 2. How serious is the problem? Why does it matter? The reduction of microbial pathogens in food products is the most pressing food safety problem today. Research is needed to elucidate the
fundamentals of growth and survival of pathogens in situ, including their resistance to sanitation practices. The pathogen reduction technologies in this research are focused primarily on post slaughter processing. Providing cost-effective improvements in plant sanitation methods will enhance the value of agricultural products for consumers as well as for the poultry industry. Agencies in need of this research include regulatory agencies, such as FSIS, the Animal and Plant Health Inspection Service, and the Environmental Protection Agency (EPA). NSF and the 3-A Symbol Council (3-A) have requested data from this research for setting standards for equipment production. Commercial interests include poultry processors, chemical and equipment manufacturers, and consumer interests such as the US Poultry and Egg Association and National Chicken Council. 3. How does it relate to the national Program(s) and National Program Component(s) to which it has been assigned? This project relates to
the ARS National Program Action Plan?(108) Food Safety and Animal Products. The research seeks to improve programs for the American public's health, safety, and well-being by providing knowledge to determine the fundamental parameters of microbial interactions with processing surfaces. Specifically, the project falls into categories 2.1.1.6, 2.2.1.5, and 2.2.1.6 of the National Program (108) Action Plan. 4. What was your most significant accomplishment this past year? A. Single Most Significant Accomplishment during FY 2002: Bacterial isolates from poultry processing were tested for susceptibility to antibiotics commonly used in therapeutic treatment of poultry. The resistance (phenotypic) properties of individual bacteria isolated from microbial consortia were characterized by PP&MQ RU scientist and support scientist. Knowledge of the antibiotic resistance patterns will aid in selection or inhibition of these bacterial species for further study. The results show that resistance to
antibiotics used for therapeutic treatment of poultry occurs in bacteria in the processing environment. B. Other Significant Accomplishment(s), if any. 1)Testing of technology to transfer a strong negative electrostatic charge to biofilms on stainless steel has shown promise to reduce bacterial contamination on surfaces. An electrostatic space charge system was used by the PP&MQ RU scientist to treat mixed bacterial populations from the poultry processing environment. The negative air ionization system effectively decreased the survival levels of bacteria on stainless steel coupons with a 99.8% kill, and is now being tested on bacterial spores. This technology could have applications that extend into other food processing areas, medical institutions, and the home. 2)For many years, the use of rubber fingers on mechanical pickers to remove feathers from broilers after scalding was considered a major contributor to cross-contamination. Studies on natural populations in the processing
plant began by the PP&MQ RU scientist testing rubber picker fingers that had been in use in commercial processing equipment. Preliminary data from three processing plants indicated that bacterial counts were much lower than expected, even lower than on fingers exposed in the laboratory to bacterial cultures. Final results may impact the schedule for replacing picker fingers and help reduce contamination and cross-contamination in the picker room. 3)Effective control or intervention measures to improve sanitation practices for bacterial contamination in processing facilities are needed. This problem has been addressed by studying the efficacy of potential chemical agents selected from several groups of substances that destroy or limit microbial growth under a CRADA with AJAY North America, LTD. The selected test compound was effective against bacterial pathogens in the laboratory and on processing surfaces. The three-year term of the CRADA ended 3/14/02. C. Significant Activities
that Support Special Target Populations. None D. Progress Report. None 5. Describe your major accomplishments over the life of the project, including their predicted or actual impact? This is the first year of a new research project. In the previous project, methods were developed to measure attached bacteria and biofilm formation on surfaces, surface materials that were resistant to bacterial attachment were identified, and physical and electrochemical treatments of stainless steel were assessed for inhibition of bacterial attachment and biofilms. Atomic force microscopy was shown to be a useful tool to predict the potential for bacteria to attach and form biofilms on surfaces. Factors that are important for resistance of stainless steel surfaces, the most common in food processing, to bacterial contamination were determined. Optimal stainless steel treatments for simultaneously preventing biofilm development and corrosion effects were determined. The efficacy of new on-line
technology for reduction of fecal contamination on broiler carcasses was tested. Digital aroma technology was put to novel use to detect, compare, profile, and classify the characteristics of bacterial species important to poultry processing as potential pathogens. The substrate utilization profiles of bacterial communities (biofilm) were assessed as they developed on chicken meat samples stored for varying time periods at two temperatures commonly used in a poultry processing facility. Research on the incidence and level of Campylobacter jejuni in the poultry processing environment was initiated. 6. What do you expect to accomplish, year by year, over the next 3 years? This research is directed to reduce pathogen contamination in food products by developing methodologies to detect, identify, quantify, prevent, and control target pathogens. FY2003-Optimum treatment parameters and reduction efficiency will be determined for the Electrostatic Space Charge System. FY2004-Bacteriological
methods will be developed and applied to generate experimental models for biofilms containing pathogens. Optimum surface treatments that combine control strategies will be determined. FY2005-Experimental model will be used to evaluate the existence of target pathogens within biofilms. 7. What technologies have been transferred and to whom? When is the technology likely to become available to the end user (industry, farmer other scientist)? What are the constraints, if known, to the adoption durability of the technology? Nothing to report for this period. 8. List your most important publications and presentations, and articles written about your work (NOTE: this does not replace your review publications which are listed below) Articles written about the work: From Farm to Fork. 2002. The American Legion Magazine. April issue. p. 39- 42.
Impacts
(N/A)
Publications
- Arnold, J.W., Mitchell, B.W. Use of negative air ionization for reducing microbial contamination on stainless steel surfaces. Journal of Applied Poultry Research. 2002. v. 11. p. 179-186.
- Boothe, D.H., Arnold, J.W. Electronic nose analysis of volatile compounds from poultry meat samples, fresh and after refrigerated storage. Journal of the Science of Food and Agriculture. 2001. v. 82. p. 315-322.
- Arnold, J.W., Boothe, D.H. Antibiotic resistance profiles of bacterial isolates from poultry products. American Society for Microbiology. Birmingham, Alabama. 2001. Proceedings Abstracts p. 40.
- Arnold, J.W. Prevention of bacterial fouling on food equipment surfaces. American Oil Chemists Society. Montreal, Canada. 2002. AOCS Annual Meeting. Abstracts p. S51.
- Boothe, D.H., Arnold, J.W. Nutrient substrates used by bacteria from the poultry processing environment. 2002. Poultry Science. v. 81(Suppl.1): Abstract #163.
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