Progress 07/01/03 to 06/30/09
Outputs
OUTPUTS: A direct colony immunoblot method (DGI) for the enumeration of Vibrio vulnificus was develped. Over the course of this project the results produced 34 abstracts that were presented at international and national meetings. As a result of work done on this project Dr. Janes organized and moderated 6 symposia on seafood safety. Dr. Janes was invited to give 7 presentations related to the results of this project. PARTICIPANTS: During this project a total of 6 M.S. students and 7 Ph. D. students graduated under my supervision. TARGET AUDIENCES: The newly developed antibody based rapid methods could be used by regulatory agencies for detection of this pathogen in oysters and the oyster industry. The data generated on the strain-to-strain differences of V. vulnificus and V. parahaemolyticus to survive at various temperatures will be useful in updating FDA risk assessment models for these pathogens. Our results have shown that acidified sodium chlorite is effective in inhibiting the growth of L. monocytogenes when this pathogen was grown on the surface of ready-to-eat meat products and have benefit Louisiana based ready-to-eat meat processing companies. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Objective 1: Over the course of this project we have developed several antibody based methods for the detection and enumeration of V. vulnificus and V. parahaemolyticus in oysters. A direct colony immunoblot method (DCI) for the enumeration of Vibrio vulnificus was developed. The DCI was then compared to the DNA hybridization procedure (DNAH). Both DCI and DNAH detected 1-2 log colony forming units (CFU)/ml V. vulnificus mixed with 4 log CFU/ml V. parahaemolyticus. Both methods were comparable and demonstrated no significant statistical differences. An immune magnetic separation protocol was developed using anti-H antibody for the concentration of V. vulnificus from phosphate buffered saline (PBS) suspensions and spiked oyster homogenate. Immunomagnetic separation (IMS) with V. vulnificus-spiked PBS yielded a binding percentage ranging from 19 to 57 percent while IMS with spiked oyster homogenate carried out at 2 different V. vulnificus concentrations exhibited a percentage binding of 25 to 57 percent. Our research has shown that various V. vulnificus and V. parahaemolyticus strains vary significantly in their ability to survive and grow at refrigeration temperatures. Some strains showed increased survival rates throughout the experimental storage times and temperatures investigated, while others had significantly lower growth or survival. Results from this study have shown differences in adaptation responses between the various Vibrio vulnificus and some Vibrio parahaemolyticus strains that were cold temperature adapted at 15degreeC. The cold adaptation response was more sustained for the V. vulnificus strains at some temperatures tested, while for the V. parahaemolyticus strains exhibited a response but this response was generally short lived. Objective 2: With the recurring recalls of ready-to-eat meat and poultry products due to contamination by Listeria monocytogenes, there is a clear need to develop additional methods to prevent economic loss and possible deaths that can occur from foodborne listeriosis infections. Our results have shown that acidified sodium chlorite is effective in inhibiting the growth of L. monocytogenes when this pathogen was grown on the surface of ready-to-eat meat products and is beneficial to Louisiana-based ready-to-eat meat processing companies. Objective 3: We have collected fecal samples from two agricultural research stations in Louisiana and found a 4.9 percent possible prevalence (8 positive out of 164 cattle tested) of E. coli O157:H7 in the cattle. This study has shown that biofilms from cattle water troughs can support the growth of L. monocytogenes, E. coli O157:H7 and S. typhimurim. These results suggest that methods need to be developed to control these pathogens in biofilms in cattle water troughs. Objective 4: Edible films and coatings containing antimicrobial agents have the potential to control foodborne pathogens on the surface of food products. The promising results of our research demonstrated that chitosan could possibly be used as an antimicrobial coating on the surface of lettuce to reduce the risk of E. coli O157:H7 contamination.
Publications
- V. E. Burnham , M. E. Janes, L. A. Jakus, J. Supan, A. DePaola and J. Bell. 2009. Growth and survival differences of Vibrio vulnificus and Vibrio parahaemolyticus strains during cold storage. Journal of Food Science 74:M314-M318.
- Stephenie L. Drake, Richelle Beverely, Amrish Chawla, Marlene Janes, John Supan, Jon Bell, Jay F. Levine, and Lee-Ann Jaykus. 2009. A simplified method to monitor internal oyster meat temperature on a commercial scale. Food Prot. Trends. May.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: The results of this project were presented at the IAFP meeting in 2008 and the title was "Isolation of Vibrio vulnificus from oyster homogenate by immunomagnetic separation using anti-H monoclonal antibodies." Furthermore, a provisional patent was applied for by the LSU Ag Center in 2008 titled "Detection of Vibrio vulnificus and Vibrio parahaemolyticus via mononclonal antibodies." PARTICIPANTS: This research project fully supported one Ph.D. student Ravi Jadeja who will graduate in December 2010. Dr. Janes was the lead PI on the grant and assisted with the design of the research project, supervision of the student, and analysis of the data. Dr. Janet Simonson was a Co-PI and assisted with the production of the antibodies and design of the research project. TARGET AUDIENCES: The IMB coated with monoclonal antibody specific for V. vunificus could be combined with Real-time PCR for extraction of V. vunificus from oyster tissue to remove inhibitors that can interfere with PCR. This technique could lead to a simple and rapid method by Regulator agencies for detection of this pathogen in oysters. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Immunomagnetic beads (IMB) coated with monoclonal antibody specific for V. vunificus could prove useful for the isolation and concentration of the organism from complex environmental samples. This study was aimed at developing an immunomagnetic separation protocol using anti-H monoclonal antibody for the recovery of V. vunificus from spiked oyster homogenates. Two different sets of IMB were prepared by mixing sheep anti-mouse IgG IMB with monoclonal antibodies MAB 8-D-4 and MAB 3-D-10 reactive with V. vunificus flagellar core at a concentration of 5 micro-grams IgG/10,000,000 IMB. The binding capacity of the beads coated with monoclonal antibodies was determined by incubating 10,000,000 IMB with 500 micro-liters spiked oyster homogenate on a shaker at 25degreeC for 30 minutes followed by separation with an immunomagnetic bead concentrator. The number of unbound bacteria was determined by plating the aspirated supernatant fluid on TCBS and TSA plus 2percent NaCl agar plates. Three strains of V. vulnificus (strain 1007, C7164, and ATCC 27562) were used during this study with two different concentrations 100 and 1000 V. vulnificus/ml. At 1000 CFU/ml mean percentage binding of V. vulnificus 1007 with MAB 8-D-4 was determined to be the highest at 57percent, followed by V. vulnificus C7164 at 52percent and V. vulnificus ATCC 27562 at 35percent which is significantly lower than the other two strains. At 100 CFU/ml the binding percentage for three strains was about 25percent with MAB 8-D-4. MAB 3-D-10 at 1000 V. vulnificus/ml showed the highest percentage mean binding of 56percent for V. vulnificus 1007, followed by V. vulnificus C 7164 with 52percent while V. vulnificus ATCC 27562 had 35percent. The binding percentages at 100 CFU/ml for V. vulnificus C7164, V. vulnificus ATCC 27562 and V. vulnificus 1007 were about 28percent. Difference between the percentage binding of ATCC 27562, 1007 and C7164 might be due to non-motile cells present in the ATCC strain.
Publications
- R. Senevirathne, M.E. Janes and J. Simonson. 2008. Detection and enumeration of Vibrio vulnificus by direct colony immunoblot. Journal of Food Science, 74:M41-M45.
- R. Beverly, M. E. Janes, W. Prinyawiwatkul, and H. K. No. 2008. Edible chitosan films on ready-to-eat roast beef for the control of Listeria monocytogenes. Food Microbiology 25: 534-537.
- K. Melody, R. Senevirathne, M. Janes, L. A. Jaykus, and J. Supan. 2008. Effectiveness of Icing as a Post-Harvest Treatment for Control of Vibrio vulnificus and V. parahaemolyticus in the eastern oyster (Crassostrea virginicA). Journal of Food Protection, 71:1475-1480.
- S. Seo, J.M. King, W. Prinyawiwatkul, and M.E. Janes. 2008. Antibacterial activity of ozone-deolymerized crawfish chitosan. Journal of Food Science 73:M400-M404.
- S. Datta, M. E. Janes and J. G. Simonson. 2008. Immunomagnetic separation and coagglutination of Vibrio parahaemolyticus with anti-flagellar monoclonal antibody. Clinical and Vaccine Immunology, 15:1541-1546.
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: In 2007 we presented a poster at the International Association for Food Protection meeting title "Comparison of the direct colony immunoblot to DNA hybridization for enumeration of Vibrio vulnificus in oysters." From this meeting several companies were interested in licensing the antibodies.
PARTICIPANTS: Project title: Adaptation differences of Vibrio vulnificus strains during cold storage. This research project has fully supported one M.S. student Veronica Burnham who graduated in August 2006. Dr. Lee-Ann Jaykus from University of North Carolina was the lead PI on the grant and assisted with the design of the research project. Dr. Marlene Janes was the lead PI for this research project and assisted with the design of the project, supervision of the student and analysis of the data. Project title: Detection of Vibrio vulnificus by direct colony immunoblot. This research project has fully supported one M.S. student Ms. Reshani N. Senevirathne and partially supported a Ph. D student Ms. Shreya Datta. Ms. Reshani Senevirathne won the 2006 Tom Quinn IFT student scholarship for her research project "Direct colony immunoblot for enumeration of Vibrio vulnificus." Shreya Datta helped with the production of the monoclonal antibodies. Dr. Janet Siomonson Co-PI from the LSU Ag
Center assisted with the production of the monoclonal antibodies and the development of the immunoblot. Dr. Marlene Janes the lead PI for the project managed the project and supervised the graduate students.
TARGET AUDIENCES: Federal agencies can use this data for updating risk assessment models for these pathogens. The newly developed direct colony immunoblot for detection of V. vulnificus could be used as a rapid enumeration method by regulatory agencies or the seafood industry.
Impacts
At 5degreeC by day 10, V. vulificus strain 515-4C2 had the highest counts, with 1.97 log CFU/ml, strain 541(O) 49C had counts of 0.65 log CFU/ml while strain 33815 reached non-detectable levels at day 8. At 8degreeC by day 10 V. vulnificus Strain 515-4C2 had the highest counts, with 2.23 log CFU/ml, strain 29306 had counts of 1.15 log CFU/ml while strain 33815 reached non-detectable levels. At 10degreeC by day 10 V. vulnificus Strain 541(O) 49C had the highest counts, with 8.02 log CFU/ml, strain 0106-14 had counts of 7.00 log CFU/ml while strain 33815 reached non-detectable levels. V. vulnificus strains cultured at 37degreeC and then held at 15degreeC (cold temperature adaptation for four hours, before storage at 5 or 8degreeC for 9 days, exhibited better survival rates (but no growth) than cultures transferred directly from 37degreeC to 5 or 8degreeC. Cultures similarly held at 15degreeC before storage at 10degreeC resulted in better growth rates than cultures
transferred directly to 10degreeC. The duration of either the growth or survival responses differed for various V. vulnificus strains. Some strains showed increased survival rates throughout the experimental storage times and temperatures investigated, while others exhibited significant increases in growth or survival for shorter time periods. Our research has shown that various V. vulnificus strains vary significantly in their ability to survive and grow at refrigeration temperatures. These data will be useful in updating risk assessment models for these pathogens. We have developed and optimized a V. vulnificus direct colony immunoblot method that is user-friendly in that it requires no cell lysis, less manipulations, and less stringent temperature requirements. Our newly developed V. vulnificus direct colony immunobolt can consistently detect on selective agar plates, that under normal counting procedures would be to numerous to count, 100 CFU/ml of V. vulnificus that was mixed
with 10,000 CFU/ml of V. parahaemolyticus. The direct colony immunoblot was positive for all V. vulnificus strains tested and did not cross react with other Vibrio species tested. Detection of V. vulnificus by the direct colony immunoblot or DNA probe methods was comparable with no significant statistical difference. The direct colony immunoblot method had better color development and was less time-consuming than the DNA probe method. Our V. vulnificus direct colony immunoblot method only takes 3 hours while the DNA probe hybridization method takes 24 h. The newly developed direct colony immunoblot for detection of V. vulnificus could possibly be used as a rapid enumeration method by regulatory agencies or the seafood industry.
Publications
- S. Datta, M. E. Janes, Q-G. Xue, J. Losso, and J. F. LaPeyre. 2008. Control of Listeria monocytogenes and Salmonella anatum on the Surface of Smoked Salmon Coated with Calcium Alginate Coating containing Oyster Lysozyme and Nisin. 73:M67-M71.
- A. Chawla, J.W. Bell, and M.E. Janes. 2007. Optimization of ozonated water treatment of wild-caught and mechanically peeled shrimp meat. J. Aquatic Food Product technology. 16:41-56.
- F. Han, R.D. Walker, M.E. Janes, W. Prinyawiwatkul, and B. Ge. 2007. Antimicrobial susceptibilities of V. parahaemolyticus and V. vulnificus isolates from Louisiana Gulf and retail raw oysters. Applied Environ. Micro. 73:7096-7098.
- Ting, S., Z. Xu, C.T. Wu, M.E. Janes, W. Prinyawiwatkul, and H.K. No. 2007. Antioxidant activities of different colored sweet bell peppers (Capsicum annuum L.) Journal of Food Science 72:98-102.
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Progress 01/01/06 to 12/31/06
Outputs
Project 1: Vibrio vulnificus and V. parahaemolyticus are the most common Vibrios associated with seafood illness in the United States. Our study was conducted to determine if strain-to-strain differences exist in the growth and survival of eight different V. vulnificus and V. parahaemolyticus strains at low temperatures and if eight different V. vulnificus and V. parahaemolyticus strains had different cold temperature adaptation responses. At 5 or 8degreeC by day 10 V. vulificus strain 515-4C2 had the highest counts while strain 33815 reached non-detectable levels. At 10degreeC by day 10 V. vulnificus strain 541(O) 49C had the highest counts while strain 33815 reached non-detectable levels. V. vulnificus strains cultured at 37degreeC and then held at 15degreeC for 4h (cold temperature adaptation), before storage at 5, 8 or 10degreeC for 9 days, exhibited better survival rates than cultures transferred directly from 37degreeC to 5, 8 or 10degreeC. At 5degreeC by day 10
V. parahaemolyticus strain 541(O) 57C had the highest counts while strain 33847 had significantly lower counts. At 8degreeC by day 10 V. parahaemolyticus strain M350A had the highest counts with 7.97 log CFU/ml and strain 541(O) 57C had the lowest counts, with 4.80 log CFU/ml. At 10degreeC V. parahaemolyticus strain NY477 had the highest counts, with 8.31 log CFU/ml and strain 33847 had the lowest counts, with 6.77 log CFU/ml. Six of the eight V. parahaemolyticus cultures that were held at 15degreeC (cold temperature adaptation) prior to storage at 5, 8 or 10degreeC, had better survival rates than cultures transferred directly from 37degreeC to 5, 8 or 10degreeC. Project 2: E. coli O157:H7 have the ability to attach to a variety of surfaces and form biofilms that persist in the food environments and contaminate food during processing. Cetylpyridinium chloride (CPC) is frequently being used as disinfectants in food industries to prevent the spread of microorganisms. The objective of
this study was to determine the antimicrobial effects of CPC on E. coli O157:H7 biofilms grown in different growth media and temperature conditions on the surface of stainless steel. A five-strain mixture of E. coli O157:H7 (108 CFU/ml) cell suspension was prepared and attached to stainless steel chips then transferred to tubes containing tryptic soy broth (TSB), 10% TSB, or minimal salts broth (MSB) and incubated at 12degreeC and 22degreeC for 2, 10 and 21 days. The biofilms were treated with 0.01%, 0.05%, 0.5%, 1.0% CPC solutions for 2, 5, and 10 min. The inactivations of biofilms grown in all the three media were time and concentration dependent. The 21-day and 10-day biofilms were significantly (P≤ 0.05) more resistant to CPC than 2-day biofilms. Biofilms grown in MSB were more resistant to CPC than biofilms grown in TSB or 10% TSB. Concentrations of 0.05% and 0.1% were sufficient to completely inactivate biofilms grown in TSB and 10% TSB, respectively; whereas, a
concentration of 1.0% CPC was required to completely inactivate the 21-day biofilms grown in MSB.
Impacts
Project 1: Our research has shown that various V. vulnificus and V. parahaemolyticus strains vary significantly in their ability to survive and grow at refrigeration temperatures. Some strains showed increased survival rates throughout the experimental storage times and temperatures investigated, while others had significantly lower growth or survival. Our results from this study have shown differences in adaptation responses between the various Vibio vulnificus and some Vibrio parahaemolyticus strains that were cold temperature adapted at 15degreeC. The cold adaptation response was more sustained for the V. vulnificus strains at some temperatures tested, while for the V. parahaemolyticus strains exhibited a response but this response was generally short lived. This data may be useful in updating FDA risk assessment models for these pathogens. Project 2: E .coli O157:H7 biofilms grown in MSB were more resistant to CPC than biofilms grown in TSB or 10% TSB. In general,
the inactivation's of biofilms grown in all the three media were time and concentration dependent. Longer exposure times significantly increased biocidal activity and old biofilms were more resistant to CPC sanitizing than young biofilms. This study also suggests that E. coli O157:H7 biofilms grown under low-nutrient conditions were more resistant than biofilms grown in rich media. This study shows the potential use of CPC as a sanitizer to prevent E. coli O157:H7 cross-contamination of food products by stainless steel surfaces in food processing plants.
Publications
- V.E. Burnham, M.E. Janes, R.L. Beverly, and L. Jaykus. 2006. Strain to strain differences in the growth and survival of Vibrio parahaemolyticus in broth. Presented at the IFT meeting in Orlando, Fl, June 2006, (Abstract number 3A-27)
- R.N. Senevirathne, and M.E. Janes, J. Simonson. 2006. Direct colony immunoblot for enumeration of Vibrio vulnificus. Presented at the IFT meeting in Orlando, Fl, June 2006, (Abstract number 3A-27)
- S. Plauche, and M.E. Janes. 2006. Antimicrobial effects of cetylpyridinium chloride against Listeria monocytogenes, Salmonella typhimurium and Escherichia coli O157:H7 growth on biofilms. Presented at the IFT meeting in Orlando, Fl, June 2006, (Abstract number 54A-29)
- Dupard, T., M.E. Janes, R.L. Beverly and J. Bell. 2006. Antimicrobial effect of cetylpyridinium chloride against Listeria monocytogenes growth on the surface of raw and cooked retail shrimp. Journal of Food Science 71:241-244.
- Osman, M., M.E. Janes, R. Story, R. Nannapaneni, and M.G. Johnson. 2006. Differential kill activity of cetylpyridinium chloride with or without Bacto neutralizing buffer quench against firmly adhered Salmonella Gaminara and Shigella sonnei on cut lettuce stored at 4 o C. Journal of Food Protection 69:1286-1291.
- A. Chawla, J.W. Bell, M.E. Janes and C. Pollet. 2006. Development of a Process to Measure Ozone Concentration in Processing Water at the Point of Product Application. Ozone: Science and Engineering 28:171-175.
- S. Plauche, and M.E. Janes. 2006. Prevalence of Escherichia coli O157:H7 in cattle water troughs in Louisiana. Presented at the IFT meeting in Orlando, Fl, June 2006, (Abstract number 39H-15)
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Progress 01/01/05 to 12/31/05
Outputs
Objective 1, we studied the antimicrobial effect of bacteriophage treatments against V. vulnificus virulent (opaque) and attenuate (translucent) and V. parahaemolyticus in tryptone broth incubated at 30degreeC for 24h. After 24h V. vulnificus opaque counts treated with Vvo phage were reduced by 1.95 log CFU/g, V. vulnificus translucent counts treated with Vvt phage were reduced by 2.90 log CFU/g and V. parahaemolyticus counts treated with Vp phage were reduced by 1.02 log CFU/g as compared to the control non-treated counts. Objective 2, we investigated the concentration of cetylpyridinium chloride (CPC) (0.05, 0.1, 0.2, 0.4, 0.6, 0.8 or 1.0percent) with or without a water rinse that would effectively reduced L. monocytogenes on the surfaces of raw or cooked shelled or unshelled shrimp stored at 4degreeC for 24h. All CPC concentrations with a water rinse reduced L. monocytogenes counts on the surface of cooked shrimp by about 2.5 log CFU/g. Conversely, without a water
rinse L. monocytogenes counts on the surface of cooked shrimp were reduced 3.0 log CFU/g with 0.1, 0.2 or 0.4percent CPC, 5.0 log CFU/g with 0.6percent CPC, 6.0 log CFU/g with 0.8percent CPC and 7.0 log CFU/g with 1.0 percent CPC as compared to the control non-treated samples. Objective 3, The effect of temperature and nutrients on the antimicrobial effects of copper and brass against Listeria monocytogenes were examined in Modified Welshimers broth (MWB) or MWA (agar) at 4degreeC, 25degreeC, or 37degreeC. In regular MWB, L. monocytogenes counts had reach non-detectable levels for copper and brass after 72h at 4degreeC, 8h at 25degreeC, and 6h at 37degreeC. If the concentration of glucose in MWA (10g) was reduced to 1g or increased to 20g, the antimicrobial activity of the copper and brass against L. monocytogenes was significantly reduced. However, changing the carbohydrate in regular MWA from glucose to fructose, mannose or cellobiose reduced the antimicrobial effect of copper and
brass against L. monocytogenes. Higher phosphate levels in MWA increased the antimicrobial activity of copper and brass against L. monocytogenes. Amino acids at a high or low concentration in MWA reduced the antimicrobial effective of the copper or brass against L. monocytogenes. Objective 4, this study investigated the antimicrobial effect of oyster lysozyme (OysL) and hen egg white lysozyme (HEWL) with or without nisin, added to calcium alginate coating (CaAlg) coated onto smoked salmon against Listeria monocytogenes and Salmonella anatum stored at 4degreeC for 35 d. Our results indicated that the effectiveness of oyster lysozyme or hen egg white lysozyme was enhanced when added to calcium alginate coatings. Furthermore, after 35 d at 4degreeC the growth of L. monocytogenes and S. anatum was suppressed by about 2.3 log CFU/g with CaAlgNOysL or CaAlgNHEWL coatings compared to the control non-treated samples. There was no significant difference between oyster lysozyme and hen egg
white lysozyme treatments against L. monocytogenes or S. anatum inoculated on the surface of salmon.
Impacts
Objective 1, Our results indicate that bacteriophages active against Vibrio vulnificus and Vibrio parahaemolyticus could possibly be used as a processing aid to control this pathogen. Objective 2, to date, the use of CPC has been approved by the FDA at a level not to exceed 0.3 grams of CPC and should also contain propylene glycol at a concentration of 1.5 times that of the CPC per pound of raw poultry carcass. Although the use of CPC as an antimicrobial agent for seafood has not been approved by the FDA, we have shown in this study the strong potential of cetylpyridinium chloride as a washing solution to reduce L. monocytogenes on the surface of raw and cooked shrimp. Objective 3, copper and brass metals were effective in destroying L. monocytogenes in a nutrient rich media. However, refrigerator temperatures and available nutrients can have positive or negative affects on the antimicrobial activity of copper and brass against L. monocytogenes. The method reported
here could prove helpful in this quest for controlling this pathogen in food processing environments. Objective 4, a major concern of the smoked fish industry is contamination of their products with Listeria monocytogenes and Salmonella species. Methods need to be developed to control these pathogens on the surface of fish and seafood. Our study has shown that calcium alginate coatings containing lysozyme and/or nisin could potentially be used to reduce the growth of L. monocytogenes and S. anatum on the surface of ready-to-eat smoked salmon at refrigerated temperatures.
Publications
- T.M. Dupard, M.E. Janes, and J. Bell. 2005. Antimicrobial effect of cetylpyridinium chloride against Listeria monocytogenes on the surface of shelled raw and cooked shrimp. Abstract number 89A-19, IFT meeting in New Orleans, LA, July 2005.
- R.L. Beverly and M. E. Janes. 2005. Acidified sodium chlorite treatment for inhibition of Listeria monocytogenes growth on the surface of various ready-to-eat products. Abstract number 67-1, IFT meeting in New Orleans, LA, July 2005.
- S. Datta, Q.G. Xue, M.E. Janes, J.N. Losso, and J.F. La Peyre. 2005. Purification of lysozymes from shell liquor of eastern oysters (Crassostrea virginica). Abstract number 89B-34, IFT meeting in New Orleans, LA, July 2005.
- A. Abushelaibi, M.E. Janes, and A. Khachatryan. 2005. Antimicrobial effects of copper ions against Listeria monocytogenes at different pH. Abstract number 89D-13, IFT meeting in New Orleans, LA, July 2005.
- R.L. Beverly, M.E. Janes, and W. Prinyawiwatkul. 2005. Antimicrobial effect of chitosan coatings against Listeria monocytogenes on the surface of ready-to-eat roast beef. Abstract number 89D-9, IFT meeting in New Orleans, LA, July 2005.
- A. Dumas, M.E. Janes, and J.C. Beaulieu. 2005. Effectiveness of cetylpyridinium chloride dips to reduce foodborne human pathogens (Salmonella montevideo, Shigella sonnei, and Escherichia coli) on fresh-cut cantaloupe. Abstract number 108-9, IFT meeting in New Orleans, LA, July 2005.
- A. Abushelaibi, M.E. Janes. 2005. Effect of nutrients on the antimicrobial activity of copper and brass against Listeria monocytogenes. Abstract number P3-38, IAFP meeting in Baltimore, MD, August 2005.
- R.L. Beverly and M.E. Janes. 2005. Survival of Listeria monocytogenes on ready-to-eat meat products stored at freezer temperatures under vacuum and non-vacuum packaging. Abstract number P3-03, IAFP meeting in Baltimore, MD, August 2005.
- R.L. Beverly, M.E. Janes and G. Oliver. 2006. Acidified sodium chlorite treatment for inhibition of Listeria monocytogenes growth on the surface of cooked roast beef. J. Food Prot. 64:432-435.
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Progress 01/01/04 to 12/31/04
Outputs
Objective 1, We investigated the effects of pH, temperature and salt content on the activities of bacteriophages isolated from oysters against virulent Vibrio vulnificus (Vvo), attenuated Vibrio vulnificus (Vvt) or Vibrio parahaemolyticus (Vvt). Results showed that bacteriophages Vvo, Vvt, and Vp lost activity at an acidic pH and were sensitive to elevated temperatures. The bacteriophage Vp was more salt tolerant then the phages Vvo and Vvt. Objective 2, We investigated the growth of Listeria monocytogenes, Escherichia coli O157:H7 or Salmonella typhimurim in biofilms from cattle water troughs that were placed onto glass, wood, copper, tin or rubber at 4degreesC. Plates were placed at 4degreesC and bacterial counts determined at day 0, 7, 14, 21, and 28. L. monocytogenes was able to grow to 7.0 Log on the surface of glass, rubber and tin by 28 day study. Conversely, when biofilms were placed on the surface of wood and copper L. monocytogenes counts were reduced to
non-detectable levels by day 14 for copper and day 21 for wood. E. coli O157:H7 initial counts of 7.0 Log in the biofilms on the different surfaces slowly dropped at 4degreesC. By day 28 E. coli O157:H7 counts had reached non-detectable levels in biofilms on copper, wood and tin. The E. coli O157:H7 counts were 2.50 Log for glass and 3.54 for rubber by day 28. An initial inoculation of 6.5 Log for S. typhimurim in biofilms was reduced to 3.71 Log on glass, 4.59 Log on rubber, and 3.49 Log on tin by day 28. S. typhimurim inoculated into biofilms placed on copper or wood were reduced to non-detectable levels by day 28. Objective 3, Our study evaluated the effect of temperature (4, 25, and 37degreeC) on the antimicrobial activity of copper and brass metals against L. monocytogenes. L. monocytogenes counts were determined at day 0, 2, 4, 6 or 8. Results showed that at 4degreesC L. monocytogenes counts on the surface of brass were significantly reduced by 3 Log from day 2 to 8 as compared
to the controls (4.0 Log). L. monocytogenes counts at 4degreesC on copper were reduced to non-detectable levels by day 8. At 25degreesC L. monocytogenes counts had a 6 Log reduction by day 6 when grown on the surface of copper and brass. At 37degreesC L. monocytogenes counts on the surface of the copper or brass were reduced to non detectable levels at day 8. Objective 4, We determined the antimicrobial effects of chitosan against E. coli O157:H7 (EHEC) on the surface of lettuce. A high (HMW) or low (LMW) molecular weight chitosan was dissolved into two 0.5 or 1.0% of lactic or acetic acid. The treated samples were stored at 4degreesC for 0, 2, 4, and 6 days or at 25 degreeC for 0, 1, 2, and 3 days and bacterial counts determined. Results showed EHEC was reduced to non-detectable levels on the surface of lettuce by day 6 for all chitosan coatings at 4degreesC. By day 3 at 25degreesC the most effective chitosan coatings were 0.5 or 1% acetic acid HMW or LMW that had a 3.5 to 4.0 Log
reduction in EHEC counts. The lactic acid chitosan treatments reduced EHEC counts on the surface of lettuce from 2.0 to 3.0 Log at 25degreesC by day 3.
Impacts
Objective 1, Our results indicate that bacteriophages active against the virulent Vibrio vulnificus and Vibrio parahaemolyticus are naturally found in oysters and could possibly be used as a processing aid to control this pathogen in live oysters. Objective 2, Reservoirs responsible for the colonization of cattle with foodborne pathogens are still poorly understood. Our research indicated that E. coil O157:H7, L. monocytogenes, Salmonella typhimurim is capable of growing and surviving in biofilms found in cattle water troughs and could lead to control measure to help reduce or eliminate this pathogen in cattle water troughs. Our study showed that copper could possibly be used to control these foodborne pathogens in cattle water trough biofilms. Objective 3, With the recurring recalls of ready-to-eat meat and poultry, fish, and seafood products due to contamination by L. monocytogenes there is a clear need to develop additional methods to prevent economic loss and
possible deaths that can occur from foodborne listeriosis infections. Our results indicated that copper or brass metals could possibly be used to control L. monocytogenes in hard to clean areas such as drains or air vents of the food processing environment. Objective 4, Edible films and coatings containing antimicrobial agents have the potential to control foodborne pathogens on the surface of food products. The promising results of our research demonstrated that chitosan could possibly be used as an antimicrobial coating on the surface of lettuce to reduce the risk of E. coli O157:H7 contamination.
Publications
- Richelle Beverly and Marlene E. Janes. 2004. The reduction of Listeria monocytogenes on roast beef treatment with acidified sodium chlorite. Abstract, IAFP meeting in Phoenix, AZ, August 2004.
- Aisha Abushelaibi and Marlene E. Janes. 2004. Antimicrobial effects of copper ions on the growth of Listeria monocytogenes at different temperatures. Abstract, IFT meeting in Las Vegas, NV, July 2004.
- Marlene E. Janes and A. Bond. 2004. Listeria monocytogenes and Escherichia coli O157:H7 grown in cattle water trough biofilms on various surfaces. Abstract, ASM meeting in New Orleans, LA, May 2004.
- Aisha A. Abushelaibi and Marlene E. Janes. 2004. Chitosan as an antimicrobial coating to control Escherichia coli O157:H7 on the surface of lettuce. Abstract, IAFP meeting in Phoenix, AZ, August 2004.
- Jon W. Bell, Ligia V. A. Da Silva, and Marlene E. Janes. 2004. Quality improvement of shrimp utilizing combined surfactant rinsing and ozone water treatments. Abstract, IFT meeting in Las Vegas, NV, July 2004.
- Ligia V. A. Da Silva, Marlene E. Janes and Jon Bell. 2004. Seasonal occurrence of bacteriophages active against Vibrio parahaemolyticus and Vibrio vulnificus in live oysters. Abstract, IFT meeting in Las Vegas, NV, July 2004.
- Foong Ming Koh, Marlene E. Janes, Witoon Prinyawiwatkul, and Hong Kyoon No. 2004. Antimicrobial efficacy of chitosan coating against Listeria monocytogenes in fresh salmon. Abstract, IFT meeting in Las Vegas, NV, July 2004.
- Aisha Abushelaibi and Marlene E. Janes. 2004. Antimicrobial effects of copper ions on the growth of Listeria monocytogenes at different temperatures. Abstract, IFT meeting in Las Vegas, NV, July 2004.
- Organized and Participated in a symposia titled Packaging Innovations, Safety Concerns and Seafood. Symposia, IAFP annual meeting in Phoenix, AZ, August 2004.
- M. J. Cho, R. W. Buescher, M. Johnson, and M. Janes. 2004. Inactivation of pathogenic bacteria by cucumber volatiles (E, Z)-2,6-nonadienal and (E)-2-nonenal. J. Food Prot. 67:1014-1016.
- Richelle Beverly. 2004. The Control, Survival and Growth of Listeria monocytogenes on Food Products. Dissertation: Louisiana State University and Agricultural and Mechanical College.
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Progress 01/01/03 to 12/31/03
Outputs
Objective 1, Our results indicated that the bacteriophages found in the oyster tissue were more active against the virulent strain of Vibrio vulnificus (3.93 PFU/g) compared to the attenuated V. vulnificus strain (3.00 PFU/g) and V. parahaemolyticus (2.50 PFU/g). However, the bacteriophages found in seawater were just as effective against the virulent V. vulnificus strain (4.80 PFU/g) as the attenuated V. vulnificus strain (5.06 PFU/g) whereas no phage activity against V. parahaemolyticus was observed. Objective 2, The growth of L. monocytogenes, E. coli O157:H7 and S. typhimurim in biofilms collected from cattle water troughs on various surfaces was investigated. The biofilms were inoculated with about 3.5 Log CFU/g of L. monocytogenes, E. coli O157:H7 or S. typhimurim. Our results showed that by day 8 the counts for L. monocytogenes increased to about 7 Log CFU/g, E. coli O157:H7 and S. typhimurim increased to above 9 Log CFU/g in biofilms placed onto the surface of
wood, glass, rubber, or tin. However, when grown in biofilms on the surface of copper L. monocytogenes and E. coli O157:H7 counts were at non-detectable levels from 8 to 20 days whereas S. typhimurim counts where only reduced 1.5 Log CFU/g from control levels. These results indicate that copper ions are released slowly over time and have antimicrobial effects against L. monocytogenes and E. coli O157:H7 grown in biofilms. Objective 3, Our study evaluated various concentrations of acidified sodium chlorite (ASC) needed to effectively reduce Listeria monocytogenes counts on 5-gram cubes of ready-to-eat (RTE) roast beef samples at refrigerator temperatures. Immediately following the initial inoculation (6.4 Log CFU/g) all the cooked roast beef samples treated with ASC showed significantly decreased bacterial counts compared to control non-treated samples due to the rapid antimicrobial action of ASC against L. monocytogenes. By day 28 the 500, 750, or 1000 ppm ASC treated spicy roast beef
samples had greater than 4.00 Log CFU/g reductions whereas the same concentrations of ASC treated regular roast beef samples only had about 2.5 Log CFU/g reductions in L. monocytogenes counts compared to the controls. Objective 4, We determine the inhibitory activities of ethylenediaminetetraacetic salt (EDTA) with or without zein edible films containing nisin coated onto the surfaces of raw chicken for protection against Campylobacter jejuni (CJ) at refrigerator temperatures. An initial 7.5 log CFU/g inoculum of C. jejuni on the surface of chicken dropped to about 4 log CFU/g at day 14 through 28 at 4 degrees C. EDTA alone and zein coatings containing nisin reduced C. jejuni counts to non-detectable levels on the surface of raw chicken from 14 to 28 days. The Zein coatings + EDTA and zein coatings containing nisin + EDTA reduced C. jejuni counts to non-detectable from 7 to 28 days. The C. jejuni counts on raw chicken samples coated with nisin or zein were not significantly different
from the control counts through out the 28 days. Our results indicated that zein coatings with antimicrobial agents and/or EDTA show promise for the control of Campylobacter on the surface of raw chicken.
Impacts
Objective 1, Bacteriophages have the potential to control foodborne pathogens in seafood or could be used for the indication of Vibrio vulnificus and Vibrio parahaemolyticus contamination of oysters. Our results indicate that bacteriophages active against the virulent Vibrio vulnificus and Vibrio parahaemolyticus are naturally found in oysters and could possibly be used as a processing aid to control this pathogen in live oysters. Objective 2, Reservoirs responsible for the colonization of cattle with E. coli O157:H7 are still poorly understood. Our research indicated that E. coil O157:H7 is capable of growing and surviving in biofilms found in cattle water troughs and could lead to control measures to help reduce or eliminate this pathogen in cattle water troughs. Objective 3, With the recurring recalls of ready-to-eat meat and poultry products due to contamination by Listeria monocytogenes, there is a clear need to develop additional methods to prevent economic loss
and possible deaths that can occur from foodborne listeriosis infections. Our results have shown that acidified sodium chlorite is effective in inhibiting the growth of L. monocytogenes when this pathogen was grown on the surface of cooked roast beef at refrigerator temperatures. Objective 4, Edible films and coatings containing antimicrobial agents have the potential to control foodborne pathogens on the surface of food products. The promising results of our research demonstrated that zein coatings containing nisin and/or EDTA could control Campylobacter jejuni on the surface of raw poultry.
Publications
- Marlene E. Janes. 2003. Edible films for controlling foodborne pathogens. Presented at the National Fisheries Institutes Seafood technology innovations conference in Orlando, FL, February.
- Marlene E. Janes and M. G. Johnson. 2003. Control of Campylobacter jejuni on the surface of raw chicken coated with ethylenediaminetetraacetate with or without edible zein films containing nisin. Presented at the IAFP meeting in New Orleans, LA, August 2003.
- Bwalya Lungu, Marlene E. Janes and M. G. Johnson. 2003. Partial reduction of Listeria monocytogenes on full fat turkey frankfurters held at 4 and 8 degrees C using zein coatings containing nisin and EDTA. Presented at the ASM meeting in Washington D.C., June 2003.
- Aisha Abushelaibi, David Bankston and Marlene E. Janes. 2003. The effect of temperature fluctuations on the survival of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella typhmurim inoculated on the surface of raw irradiated chicken. Presented at the IFT meeting in Chicago, IL, July 2003.
- Julie T. Nguyen, Richelle Beverly, Ligia Da Silva and Marlene E. Janes. 2003. Prevalence of foodborne pathogens in cattle at agricultural research stations in Louisiana. Presented at the IFT meeting in Chicago, IL, July 2003.
- Richelle Beverly, Witoon Prinyawiwatkul, and Marlene Janes. 2003. Antimicrobial effect of cranberry juice against Listeria monocytogenes. Presented at the IFT meeting in Chicago, IL, July 2003.
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