Progress 02/01/15 to 01/31/19
Outputs
Target Audience:? Target audiences: For the period of the entire grant, one target communities are the main producers of jalapeno, tomato (open field) and cantaloupe melon in the north-east of Mexico where the samples were collected. More broadly are Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries who stand to benefit from the knowledge generated by the project. In this reporting period, we have reached both the Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries through presentation of our work at food safety meetings at the US (International Association for Food Protection) and additional outreach by our partners in Mexico at Universidad Autonoma de Nuevo Leon. We have also mentored 32 individuals in food safety ranging from staff, faculty, masters students and undergraduate students in the US. Efforts: We have reached out to these target audiences through publication in peer reviewed journals, presentations at conferences, annual outreach with participating farm owners, and thesis, laboratory, and other professional training provided to students. We have specifically reached out to members of the produce community, and have collaborated with the hand hygiene industry, including GOJO Industries, international partners like USAID, government partners like USDA and FDA, universities including North Carolina State University and Universidad Autónoma de Nuevo León, Mexico, and have participated in US and Mexican academic-government-industry meetings including the: Annual International Association for Food Protection meetings in 2015-2018, Latin Foods 2016 meeting, Western Center for Food Safety 2015 meeting. Additionally, we have presented our work formally to the Center for Applied Nutrition, Food and Drug Administration in 2015 and to the University of Georgia Athens Center for Food Safety in 2018. This past year, we also have four publications published and four under preparation. We also provide annual trainings to thesis, laboratory, and research students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the current reporting period, a total of 32 individuals have received professional training in project management, experimental design, laboratory techniques, statistical analysis, outreach and communication, and scientific writing. Trainees included two undergraduate students, 24 graduate students, 2 professional staff, 1 administrative staff, and 3 scientists (2 faculty). How have the results been disseminated to communities of interest?The products described above, including peer reviewed publications, websites, presentations at professional conferences, theses and dissertations, and training materials have all served to disseminate research results and engage communities of interest. What do you plan to do during the next reporting period to accomplish the goals?The project is ended so there is no further period.
Impacts
What was accomplished under these goals?
Objective 1: We developed and optimized a method for viral concentration and norovirus (GI and GII) and HAV detection on produce and environmental samples which included a matrix inhibition control. We applied this two-part method (concentration and detection) to our collected produce and environmental samples. We began with 636 samples representing 85 production chains of tomato, cantaloupe, and jalapeno pepper from field production through off-farm distribution, including 7,860 units of produce, 1,026 farm worker hand rinses, 51 kg of soil, and 567 L of source and irrigation waters from ten participating farms in northern Mexico. Of these samples, we have concentrated and extracted RNA from 80% (325/404) samples: tomato, cantaloupe, jalapeno pepper, hand rinses, irrigation, and source water. We have completed norovirus GI and GII RT qPCR testing on 50 of these samples (produce, water, hands) and repeated any matrix-inhibited samples. We found that 12% (6/50) of the samples were RT-qPCR for norovirus, with irrigation water having the highest prevalence of positive samples- 21% (3/14). A majority of the norovirus positive samples were GII (5/6). We are preparing a manuscript with these results. Objective 2: We compared our results of laboratory testing for norovirus to our database of bacterial indicators (Coliforms, E. coli, Enterococcus, coliphage) and found that there was no significant association between norovirus presence and either concentration or presence of each of the microbial indicators. We completed a systematic review to identify published indicators of norovirus and hepatitis A in the agricultural environment. We began with 262 titles and after applying our inclusion and exclusion criteria, we have 15 articles that we reviewed and extracted data for our database. We have identified a variety of viral and bacterial indicators whose relationship has been analyzed against the presence of norovirus and hepatitis A on produce, soil, water, and hands. There is no clear consensus of an indicator that is consistently present when norovirus or hepatitis A is present. We also finished conducting a systematic review of the pathogen prevalence of norovirus and hepatitis A as well as other viral, bacterial, and parasitic pathogens on the farm and processing facility environment. We have collaborated with Dr. Beatriz Quinoñes at USDA to interpret the bacterial data. This manuscript was published. We have also partnered with GOJO industries to assess if visual hand contamination is associated with viral and bacterial contamination. We published a manuscript in Food Protection Trends in 2018 showing that visual hand contamination is NOT a proxy for microbial (viral and bacterial) contamination but can be a proxy for soil contamination. The Journal requested cover photographs to improve its visibility. Through this collaboration with GOJO and our colleagues at UNAL we have also assessed the impact of hand-hygiene interventions on various produce commodities. We analyzed the efficacy of the interventions is produce-specific. Through this work, we showed that hand contamination with soil and microbes differs depending on whether jalapeño, tomatoes, or melons are harvested by farmworkers. We also assessed routes of viral and bacterial contamination in the harvest and production environment. We published two papers on this showing that hand contamination, compared to soil and water, is the most important route for microbial contamination (published in Applied Environmental Microbiology [AEM]) AND that equipment surfaces may be an important route of contamination (published in Journal of Food Protection). The AEM article was represented by a photographic montage on the cover of the Journal issue. In collaboration with Drs. Santos Garcia and Norma Heredia at the Universidad Autónoma de Nuevo Leon, Mexico, we described the phylogroups, pathotypes, biofilm formation, and antimicrobial resistance of E. coli isolates from farms and packing facilities of tomato, jalapeños, and cantaloupes. This data was published in Int J Food Microbiology. We have also optimized our rinse-membrane filtration microbe detecting assays and have applied them to pathogens (Salmonella, E. coli O157:H7, Listeria) and have analyzed the laboratory data. Objective 3. We have revised our QMRA model for norovirus contamination on fresh produce in the farm environment based on reviewer comments from rejections from the Journal of Food Protection and the International Journal of Food Microbiology. We used a stochastic QMRA model developed in R with key parameters obtained from the literature to test the impact of behavioral interventions, such as hand hygiene and glove use, on norovirus contamination of fresh produce during harvest and packing production steps in the farm environment. In the absence of any behavioral interventions, the baseline model in which both the harvester and packer are infected with norovirus, results in high levels of norovirus contamination per commodity; average 7.11 log10 virions/lettuce head, average 7.06 log10 virions/tomato, average 7.02 log10 virions/cantaloupe, and average 4.96 log10 virions/raspberry. In assessing the impact of individual behavior interventions, complete hand washing compliance by infected farm workers reduced lettuce contamination by an average 1.09 log10 virions, relative to the baseline model. Complete glove use compliance reduced lettuce contamination by an average 0.78 log10 virions. Importantly, these results suggest individual interventions are insufficient to reduce the viral contamination to below an infectious dose of 18 virions. When investigating the contribution of combined behavioral interventions on the estimated norovirus risk from lettuce consumption, the greatest reduction in infection risk was seen by improving hand washing efficacy to 6-log reduction paired with complete hand washing compliance, as well as a combination of all three interventions, hand washing efficacy and compliance paired with complete glove use compliance. These results suggest the importance hand hygiene and combined behavioral interventions in the harvesting and production environment of fresh produce to reduce norovirus-contaminated fresh produce and risk to produce consumers. Objective 4. A stochastic QMRA model for four produce groups (leafy greens, melons, berries, produce from vines/stalks) was developed with Crystal Ball using literature-based data of parameters to assess a variety of scenarios were performed to determine baseline contamination risk and evaluate select Produce Rule guidelines (improved water quality, glove use, handwashing, equipment cleaning). Among the four produce groups, average baseline risk of norovirus infection per produce item was lowest for berries (8.7 x 10-4/item [SD=5.27 x 10-4]) and highest for melons (9.0 x 10-2/item [SD=5.0 x 10-2]). Glove use and handwashing demonstrated the greatest risk reduction capacity (199% and 49%, respectively) compared to baseline risk. The sensitivity analysis also suggested that worker hand norovirus concentration, across all QMRA model inputs, were most highly correlated with consumer risk of infection across all produce groups (r = 0.925 - 0.95). When all four Produce Rule guidelines were implemented, risk of infection was reduced between 337% (berries) and 409% (melon). We have translated this model to R and are collaborating with both NoroCORE and the Produce Safety Alliance to assure the parameters used and impact of analyses are relevant to the produce industry. Dissemination and outreach activities included submitted publications and presentations listed under products to academic, industry, and government stakeholders. These deliverables and outputs will contribute to both research on produce safety and activities in U.S. agriculture to minimize viral hazards on fresh fruit and vegetables.
Publications
- Type:
Journal Articles
Status:
Other
Year Published:
2019
Citation:
Prince-Guerra JL, Nace ME, Fabiszewski de Aceituno M, Bartz F, Arbogast JM, Gentry-Shields J, Jaykus LA, Heredia N, García S, Leon JS. (2019) Produce-Specific Differences in the Ability of Soap and a Two-Step Alcohol-Based Hand Sanitizer Intervention to Reduce Soil and Microbial Contamination on Farmworker Hands During Harvest. In preparation
- Type:
Journal Articles
Status:
Other
Year Published:
2019
Citation:
Prince-Guerra JL, Fabiszewski de Aceituno M, Bartz F, Jaykus LA, Heredia N, García S, Leon JS. (2019) Detection of Norovirus in Agricultural Water, Produce, and Hand-Rinse Samples from Northern Mexico. In preparation
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Bhatta RM. Marsh Z. Newman KL. Rebolledo PA. Huey M. Hall AJ, Leon JS. 2019 Norovirus Outbreaks on College and University Campuses. The Journal of American College Health. Accepted.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Lively JY, Johnson SD, Wikswo M, Gu WD, LEON J, Hall AJ. Clinical and Epidemiologic Profiles for Identifying Norovirus in Acute Gastroenteritis Outbreak Investigations. Open Forum Infectious Diseases. 2018 Apr 10; 5(4): 1-8
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Corzo-Ariyama HA, Garcia-Heredia A, Heredia N, Garcia S, Leon J, Jaykus L, Solis-Soto L. 2019. Phylogroups, pathotypes, biofilm formation and antimicrobial resistance of Escherichia coli isolates in farms and packing facilities of tomato, jalapeno pepper and cantaloupe from Northern Mexico. Int J Food Microbiol 290:96-104.
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Van Pelt AE, Quinones B, Lofgren HL, Bartz FE, Newman KL, Leon JS. 2018. Low Prevalence of Human Pathogens on Fresh Produce on Farms and in Packing Facilities: A Systematic Review. Front Public Health 6:40
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Morrill VN, Fabiszewski de Aceituno AM, Bartz FE, Heredia N, Garcia S, Shumaker DJ, Grubb J, Arbogast JW, Leon JS. 2018. Visible Soil as an Indicator of Bacteria Concentration on Farmworkers⿿ Hands. Food Protection Trends 38:122⿿128.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2018
Citation:
Nace, Molly. Efficacy of Two Hand Hygiene Interventions at Reducing Hand Contamination Among Produce Farmworkers in Northern Mexico. Masters in Public Health Epidemiology ⿿ Rollins School of Public Health. 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Nace M, Prince-Guerra J, Fabiszewski de Aceituno AM, Bartz F, Gentry-Shields J, Jaykus LA, Heredia N, Garcia S, Leon S (2018). Efficacy of Two Hand Hygiene Interventions at Reducing Hand Contamination Among Produce. International Association for Food Protection, Salt Lake City, UT, July 8, 2018.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Sobolik J, Newman K, Jaykus LA, Leon J (2018). Norovirus Contamination of Fresh Produce During Harvesting and Packing: A Quantitative Microbial Risk Assessment Model Approach. International Association for Food Protection, Salt Lake City, UT, July 8, 2018.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2018
Citation:
Newman Kira. The Acute Serum Cytokine Response to Norovirus Infection and Illness. PhD in Epidemiology ⿿ Rollins School of Public Health. 2018
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2019
Citation:
Wickson Alexandra. Understanding the relationship between norovirus and indicator organisms on predicted contamination of produce along the United States-Mexico border ⿿ Rollins School of Public Health 2019
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Leon JS, Prince-Guerra J, Bartz FE, Garcia S, Heredia N, Jaykus LA (2018). Clean Greens: Routes of Fresh Produce Bacterial and Viral Contamination and Hygiene Interventions among Farms and Packing Facilities on the U.S.-Mexico Border. University of Georgia Center for Food Safety. Athens, GA. Invited Speaker.
|
Progress 02/01/17 to 01/31/18
Outputs
Target Audience:For the period of the entire grant proposal, one target communities are the main producers of jalapeno, tomato (open field) and cantaloupe melon in the north-east of Mexico where the samples were collected. More broadly are Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries all stand to benefit from the knowledge generated by the project. In this reporting period, we have reached both the Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries through presentation of our work at food safety meetings at the US (International Association for Food Protection). We have also mentored 25 individuals in food safety ranging from staff, faculty, masters students and undergraduate students in the US. Efforts: We have specifically reached out to members of the produce community, and have collaborated with the hand hygiene industry, including GOJO Industries, international partners like USAID, government partners like USDA and FDA, universities including North Carolina State University and Universidad Autónoma de Nuevo León, Mexico, and have participated in US and Mexican academic-government-industry meetings including the: International Association for Food Protection '17 meeting. We have also three publications published (one the cover of Applied Environmental Microbiology), two accepted, and four under preparation. We also provide annual trainings to thesis, laboratory, and research students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the current reporting period, a total of 25 individuals have received professional training in project management, experimental design, laboratory techniques, statistical analysis, outreach and communication, and scientific writing. Trainees included two undergraduate students, 18 graduate students, 2 professional staff, 1 administrative staff, and 3 scientists (2 faculty) during the reporting period. How have the results been disseminated to communities of interest?The products described above, including submitted peer reviewed publications and presentations at international professional conferences have all served to disseminate research results and engage communities of interest. What do you plan to do during the next reporting period to accomplish the goals?Generally, we plan to continue work on all objectives as they are all advanced but in various stages of work: Objective 1: We plan to complete concentration and amplification of all the remaining samples. Once completed, we plan to begin an initial draft of the norovirus and HAV concentration and optimization method for publication. Objective 2: We plan to complete our systematic review of indicators of NoV and HAV and pathogen prevalence. We also plan to link our Objective 1 results to our existing database of bacterial and viral indicators for preliminary analysis. We plan to complete the pending acceptance of 2 publications (Food Protection Trends, Frontiers in Public Health) and submit 1 additional publication on produce-specific hand hygiene interventions. In collaboration with Drs. Santos Garcia and Norma Heredia, we plan to submit two additional manuscripts on field-based methods to detect bacterial pathogens and describing the phylogroups, pathotypes, biofilm formation and antimicrobial resistance of microbes detected on produce in the harvest and production environment. Objective 3: We plan to resubmit our rejected manuscript through to publication. Objective 4: We plan to collect stakeholder feedback (NoroCORE, Produce Safety Alliance) on our preliminary model and Produce Rule scenarios and update our existing QMRA model to reflect these suggestions. We then plan to incorporate Objective 1 results and develop a draft publication for peer-review.
Impacts
What was accomplished under these goals?
During the current reporting period, we have made progress on all the Objectives above as described. Objective 1: We first developed and optimized a method for viral concentration and norovirus (GI and GII) and HAV detection on produce and environmental samples which included a matrix inhibition control. We then will apply this two-part method (concentration and detection) to our collected produce and environmental samples. We began with 636 samples representing 85 production chains of tomato, cantaloupe, and jalapeno pepper from field production through off-farm distribution, including 7,860 units of produce, 1,026 farm worker hand rinses, 51 kg of soil, and 567 L of source and irrigation waters from ten participating farms in northern Mexico. Of these samples, we have concentrated 80% (325/404) samples: tomato, cantaloupe, jalapeno pepper, hand rinses, irrigation, and source water. We have also completed a first norovirus GI and GII amplification on 50 of these samples (produce, water, hands) and repeated matrix inhibited samples. We found 1 GI norovirus RNA on melon samples, 2 GII on tomato hand rinse, 1 GII on jalapeño and tomato source water, 1 GII on melon source water.We are in the process of sequencing these to confirm norovirus positivity. We will also analyze hepatitis A RNA in a similar manner. Objective 2: While we complete Objective 1 and compare our norovirus and hepatitis A laboratory results to our database of bacterial indicators (Coliforms, E. coli, Enterococcus, coliphage), we have begun a systematic review on PubMed to identify published indicators of norovirus and hepeatitis A in the production environment. We began with 297 titles and after applying our inclusion and exclusion criteria, we have 67 articles that we reviewed and extracted data for our database. We have identified a variety of viral and bacterial indicators whose relationship has been analyzed against the presence of norovirus and hepatitis A on produce, soil, water, and hands. There is no clear consensus of an indicator that is consistently present when norovirus or hepatitis A is present. We are also conducting a systematic review of the pathogen prevalence of norovirus and hepatitis A as well as other viral, bacterial, and parasitic pathogens on the farm and processing facility environment. We have collaborated with Dr. Beatriz Quinones at USDA to interpret the bacterial data. This manuscript has been accepted for publication (to be published in Frontiers in Public Health). We have also partnered with GOJO industries to assess if visual hand contamination is associated with viral and bacterial contamination. We have had a manuscript accepted (to be published in Food Protection Trends) showing that visual hand contamination is NOT a proxy for microbial (viral and bacterial) contamination but can be a proxy for soil contamination. The Journal requested cover photographs to improve its visibility. We also assessed routes of viral and bacterial contamination in the harvest and production environment. We published two papers on this showing that hand contamination, compared to soil and water, is the most important route for microbial contamination (published in Applied Environmental Microbiology [AEM]) AND that equipment surfaces may be an important route of contamination (published in Journal of Food Protection). The AEM article was represented by a photographic montage on the cover of the Journal issue. In collaboration with Drs. Santos Garcia and Norma Heredia at the Universidad Autónoma de Nuevo Leon, Mexico, we plan to extend our analysis of our database of microbes detected on fresh produce, hands, soil, and water in the harvest and production environment in northern Mexico. From this work, we are describing the phylogroups, pathotypes, biofilm formation, and antimicrobial resistance of E. coli isolates from farms and packing facilities of tomato, jalapeños, and cantaloupes. We have also optimized our rinse-membrane filtration microbe detecting assays and have applied them to pathogens (Salmonella, E. coli O157:H7, Listeria), have analyzed the laboratory data, written up the manuscript and will be submitting it. Lastly, we are also assessing the impact of hand-hygiene interventions on various produce commodities. We are analyzing whether hand washing versus alcohol-based sanitizers are effective at reducing hand contamination of grower hands manipulating jalapeños, tomatoes, and melons. A second question is whether the efficacy of the interventions is produce-specific. We plan to submit this publication by June '18. Objective 3. We have revised our QMRA model for norovirus contamination on fresh produce in the production environment based on reviewer comments from two rejections from Journal of Food Protection and International Food Microbiology. We used a stochastic QMRA model developed in R with data on key parameters obtained from the literature to test the impact of hygiene and worker health practices on norovirus contamination of produce during harvest and packing. Based on this model, contamination was greatest for the first 5 to 10 items harvested and packed by infected workers after using the restroom, declining by over 2 log10 to approximately 20 virions/cm2 by the 10th item. Complete hand washing compliance by infected produce harvesters reduced contamination by 20.2% compared to the baseline model, and complete compliance by infected packers reduced contamination by 26.3%. In addition, increases in hand washing efficacy reduced contamination; a 2-fold improvement in efficacy reduced contamination by 29.9%. The model also showed that contamination of produce declined to below a detectable concentration (i.e., 10 virions/cm2) when infected workers were shedding less than 107 norovirus virions/g stool, suggesting that workers may not require furlough until the end of all detectable norovirus shedding. Based on reviewer comments, we have modeled the use of gloves and the eventual effect on Risk based on a norovirus dose response and the USDA recommended amount of produce consumed. We expect to submit this manuscript for publication by June'18. Objective 4. A stochastic QMRA model for four produce groups (leafy greens, melons, berries, produce from vines/stalks) was developed with Crystal Ball using literature-based data of parameters such as: norovirus in irrigation and rinse water, on worker hands, and on equipment. A sensitivity analysis identified the most influential parameters. A variety of scenarios were performed to determine baseline contamination risk and evaluate select Produce Rule guidelines (improved water quality, glove use, handwashing, equipment cleaning). Among the four produce groups, average baseline risk of norovirus infection per produce item was lowest for berries (8.7 x 10-4/item [SD=5.27 x 10-4]) and highest for melons (9.0 x 10-2/item [SD=5.0 x 10-2]). Glove use and handwashing demonstrated the greatest risk reduction capacity (199% and 49%, respectively) compared to baseline risk. The sensitivity analysis also suggested that worker hand norovirus concentration, across all QMRA model inputs, were most highly correlated with consumer risk of infection across all produce groups (r = 0.925 - 0.95). When all four Produce Rule guidelines were implemented, risk of infection was reduced between 337% (berries) and 409% (melon). We have translated this model to R and are collaborating with both NoroCORE and the Produce Safety Alliance to assure the parameters used and impact of analyses are relevant to the produce industry. Dissemination and outreach activities included submitted publications and presentations listed under products to academic, industry, and government stakeholders. These deliverables and outputs will contribute to both research on produce safety and activities in U.S. agriculture to minimize viral hazards on fresh fruit and vegetables.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Newman KL, Bartz FE, Johnston L, Moe CL, Jaykus LA, LEON JS. Microbial Load of Fresh Produce and Paired Equipment Surfaces in Packing Facilities Near the U.S. and Mexico Border. J Food Prot. 2017 Apr;80(4):582-589.
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Bartz FE, Lickness JS, Heredia N, Fabiszewski de Aceituno A, Newman KL, Hodge DW, Jaykus LA, Garc�a S, Leon JS. Contamination of Fresh Produce by Microbial Indicators on Farms and in Packing Facilities: Elucidation of Environmental Routes. Appl Environ Microbiol. 2017 May 17;83(11).
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Foote AM, Woods E, Fredes F, LEON JS. Rendering fecal waste safe for reuse via a cost-effective solar concentrator. Journal of Water Sanitation and Hygiene for Development 2017 June; 7 (2): 252-259
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Morril V, Fabiszewski de Aceituno A, Bartz F, Heredia N, Garcia S, Shumaker DJ, Grubb JE, Arbogast JW, LEON JS (2018). Visible "Soil" as an Indicator of Bacterial Concentration on Farmworkers Hands. Accepted by Food Protection Trends.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Van Pelt AE, Qui�ones B, Lofgren HL, Bartz FE, Newman JL, LEON JS. Low Prevalence of Human Pathogens on Fresh Produce on Farms and in Packing Facilities: A Systematic Review Accepted Frontiers in Public Health.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Tijerina-Rodr�guez LE, Heredia N, Sol�s-Soto L, LEON JS, Jaykus LA, Garcia S. Validation of a rinse-membrane filtration method for detection of Salmonella, E. coli O157:H7, Listeria on fresh produce. In preparation.
- Type:
Journal Articles
Status:
Other
Year Published:
2018
Citation:
Corzo-Ariyama HA, Garc�a-Heredia A, Heredia N, Garcia S, Leon JS, Jaykus LA, Sol�s-Soto L. Phylogroups, pathotypes, biofilm formation and antimicrobial resistance of Escherichia coli isolates in farms and packing facilities of tomato, jalape�o pepper and cantaloupe from Northern Mexico
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2017
Citation:
Berendes, David. Taking care of the backside: Aspects of the sanitation chain beyond the household toilet and their associations with fecal contamination in the public and private domains and enteric infection risk in children. PhD in Environmental Health - Rollins School of Public Health
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Prince-Guerra J, Fabiszewski de Aceituno A, Jaykus LA, Marsh Z, Talekar S, Bartz F, Heredia N, Garcia S, LEON J (2017). Detection of Norovirus in Agricultural Water, Produce, and Hand Rinse Samples from Northern Mexico International Association for Food Protection, Tampa, FL. July 9, 2017.
- Type:
Journal Articles
Status:
Other
Year Published:
2018
Citation:
Newman KL, Sobolik J, Jaykus LA, LEON JS. (2018) Norovirus transmission during produce harvest and packing: A quantitative microbial risk assessment modeling approach. In preparation.
- Type:
Journal Articles
Status:
Other
Year Published:
2018
Citation:
Marsh Z, Newman KL, Sobolik J, Jaykus LA, LEON JS. (2018) The Development of a Quantitative Microbial Risk Assessment Model to Evaluate the Efficacy of Produce Rule Interventions to Reduce Norovirus and Hepatitis A Virus Contamination of Fresh Produce on Farms and Packing Facilities. In preparation.
|
Progress 02/01/16 to 01/31/17
Outputs
Target Audience:Target audiences: For the period of the entire grant proposal, one target communities are the main producers of jalapeno, tomato (open field) and cantaloupe melon in the north-east of Mexico where the samples were collected. More broadly are Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries all stand to benefit from the knowledge generated by the project. In this reporting period, we have reached both the Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries through presentation of our work at food safety meetings in both the US (International Association for Food Protecion) and Mexico (Latin Foods 2016). We have also mentored 21 individuals in food safety ranging from staff, faculty, masters students and undergraduate students in the US. Efforts: We have specifically reached out to members of the produce community, and have collaborated with the hand hygiene industry, including GOJO Industries, international partners like USAID, and have participated in US and Mexican academic-government-industry meetings including the: International Association for Food Protection '16 meeting and the Latin Foods '16 meeting. We have also submitted one publication to Applied Environmental Microbiology and one publication to Journal of Food Protection. We also provide annual trainings to thesis, laboratory, and research students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the current reporting period, a total of 21 individuals have received professional training in project management, experimental design, laboratory techniques, statistical analysis, outreach and communication, and scientific writing. Trainees included two undergraduate students, 13 graduate students, 3 technical staff, and 3 scientists (2 faculty) during the reporting period. One MPH student obtained a 25K NoroCORE fellowship on this work thanks to his preliminary work on this project and was nominated to one of 5 best MPH theses at the Rollins School of Public Health. How have the results been disseminated to communities of interest?The products described above, including submitted peer reviewed publications and presentations at international professional conferences have all served to disseminate research results and engage communities of interest. What do you plan to do during the next reporting period to accomplish the goals?Generally, we plan to continue work on all objectives as they are all advanced but in various stages of work: Objective 1: We plan to complete concentration and amplification of all the remaining samples. Once completed, we plan to begin an initial draft of the NoV and HAV concentration and optimization method for publication. Objective 2: We plan to complete our systematic review of indicators of NoV and HAV and pathogen prevalence. We also plan to link our Objective 1 results to our existing database of bacterial and viral indicators for preliminary analysis. Objective 3: We plan to resubmit our rejected manuscript through to publication and expand our model to HAV and data from Objective 1. Objective 4: We plan to collect stakeholder feedback on our preliminary model and Produce Rule scenarios and update our existing QMRA model to reflect these suggestions. We then plan to incorporate Objective 1 results and develop a draft publication for peer-review.
Impacts
What was accomplished under these goals?
During the current reporting period, we have made progress on all the Objectives above as described. Objective 1: We first developed and optimized a method for viral concentration and NoV (GI and GII) and HAV detection on produce and environmental samples which included a matrix inhibition control. This work was presented at IAFP'16 as a poster presentation. We then will apply this two-part method (concentration and detection) to our collected produce and environmental samples. We began with 636 samples representing 85 production chains of tomato, cantaloupe, and jalapeno pepper from field production through off-farm distribution, including 7,860 units of produce, 1,026 farm worker hand rinses, 51 kg of soil, and 567 L of source and irrigation waters from ten participating farms in northern Mexico. Of these samples, we have concentrated 80% (325/404) samples: tomato, cantaloupe, jalapeno pepper, hand rinses, irrigation, and source water. We have also completed a first NoV GI and GII amplification on 50 of these samples (produce, water, hands) and repeated matrix inhibited samples. We found 1 GI norovirus RNA on melon samples, 2 GII on tomato hand rinse, 1 GII on jalapeño and tomato source water, 1 GII on melon source water, .We are in the process of sequencing these to confirm norovirus positivity. We will also analyze hepatitis A RNA in a similar manner. Objective 2: While we complete Objective 1 and compare our norovirus and hepatitis A laboratory results to our database of bacterial indicators (Coliforms, E. coli, Enterococcus, coliphage), we have begun a systematic review on PubMed to identify published indicators of norovirus and hepeatitis A in the production environment. We began with 297 titles and after applying our inclusion and exclusion criteria, we have 67 articles that we reviewed and extracted data for our database. We have identified a variety of viral and bacterial indicators whose relationship has been analyzed against the presence of norovirus and hepatitis A on produce, soil, water, and hands. There is no clear consensus of an indicator that is consistently present when norovirus or hepatitis A is present. We are also conducting a systematic review of the pathogen prevalence of norovirus and hepatitis A as well as other viral, bacterial, and parasitic pathogens on the farm and processing facility environment. We have completed data extraction from BLANK articles collected from PubMed, Web of Science, and EMBASE. We found a range of BLANK to BLANK of pathogens (bacterial, viral, parasitic) on BLANK number of fruit and vegetable commodities. One obstacle is the analysis of pathogen concentration data since this varies based on pathogen used and laboratory technique. We are collaborating with Dr. Beatriz Quinones at USDA to develop the best analysis methods for this question. We are in the process of completing this analysis for submission as a peer reviewer manuscript. Objective 3. We have completed a preliminary QMRA model for NoV contamination on fresh produce in the production environment. This was presented at IAFP'15 and a publication was rejected by the Journal of Food Protection. We are in the process of revising and resubmitting this publication. We used a stochastic QMRA model developed in R with data on key parameters obtained from the literature to test the impact of hygiene and worker health practices on NoV contamination of produce during harvest and packing. Based on this model, contamination was greatest for the first 5 to 10 items harvested and packed by infected workers after using the restroom, declining by over 2 log10 to approximately 20 virions/cm2 by the 10th item. Complete hand washing compliance by infected produce harvesters reduced contamination by 20.2% compared to the baseline model, and complete compliance by infected packers reduced contamination by 26.3%. In addition, increases in hand washing efficacy reduced contamination; a 2-fold improvement in efficacy reduced contamination by 29.9%. The model also showed that contamination of produce declined to below a detectable concentration (i.e., 10 virions/cm2) when infected workers were shedding less than 107 NoV virions/g stool, suggesting that workers may not require furlough until the end of all detectable NoV shedding. We are planning to extend this to HAV as well. Objective 4. A stochastic QMRA model for four produce groups (leafy greens, melons, berries, produce from vines/stalks) was developed with Crystal Ball using literature-based data of parameters such as: NoV in irrigation and rinse water, on worker hands, and on equipment. A sensitivity analysis identified the most influential parameters. A variety of scenarios were performed to determine baseline contamination risk and evaluate select Produce Rule guidelines (improved water quality, glove use, handwashing, equipment cleaning). Among the four produce groups, average baseline risk of NoV infection per produce item was lowest for berries (8.7 x 10-4/item [SD=5.27 x 10-4]) and highest for melons (9.0 x 10-2/item [SD=5.0 x 10-2]). Glove use and handwashing demonstrated the greatest risk reduction capacity (199% and 49%, respectively) compared to baseline risk. The sensitivity analysis also suggested that worker hand NoV concentration, across all QMRA model inputs, were most highly correlated with consumer risk of infection across all produce groups (r = 0.925 - 0.95). When all four Produce Rule guidelines were implemented, risk of infection was reduced between 337% (berries) and 409% (melon). We plan to incorporate data from Objectives 1-2 and share these preliminary results with stakeholders (CDC, FDA, produce industry groups) for feedback and refinement. One MPH student, Zach Marsh, obtained a 25K NoroCORE fellowship on this work thanks to his preliminary work on this project and completed his MPH thesis on this topic. His thesis was nominated to one of top 5 MPH theses in the Rollins School of Public Health. This work was presented at the USDA PD meeting at IAFP '16. We are in the process of converting the thesis for publication to Foodborne Environmental Virology. Dissemination and outreach activities included submitted publications and presentations listed under products to academic, industry, and government stakeholders. These deliverables and outputs will contribute to both research on produce safety and activities in U.S. agriculture to minimize viral hazards on fresh fruit and vegetables.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Fabiszewski de Aceituno A, Heredia N, Stern A, Bartz F, Venegas F, Solis-Soto L, Gentry-Shields J, Jaykus LA, LEON JS, Garcias S (2016). Efficacy of Two Hygiene Methods to Reduce Soil and Microbial Contamination on Farmworker Hands During Harvest. Food Control. Food Control Volume: 59 Pages: 787-792
- Type:
Journal Articles
Status:
Submitted
Year Published:
2016
Citation:
Microbial indicator contamination of produce and association with contamination in the farm environment in northern Mexico (2016). Bartz FE, Lickness JS, Aceituno A, Heredia N, Newman KL, Watson D, Jaykus LA, Garcia S, Leon JS. Submitted to Applied Environmental Microbiology.
- Type:
Journal Articles
Status:
Submitted
Year Published:
2016
Citation:
Newman KL, Bartz FE, Mues KE, Johnston L, Moe CL, Jaykus LA, Leon JS (2016) The Correlations Between Microbial Indicator Concentrations on Produce Packing Equipment Surfaces and Fresh Produce Collected Near the U.S. and Mexico Border. Submitted to Journal of Food Protection.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Heredia N, Caballero C, Cardenas C, Molina K, Garcia R, Solis L, Burrowes V, Bartz FE, Fabiszewski de Aceituno A, Jaykus LA, Garcias S, LEON JS (2016) Microbial Indicator Profiling of Fresh Produce and Environmental Samples from Farms and Packing Facilities in Northern Mexico. Journal of Food Protection Vol. 79, No. 7, 2016, Pages 11971209
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Bartz FE, Hodge DW, Heredia N, de Aceituno AF, Sol�s L, Jaykus LA, Garcia S, LEON JS. (2016) Somatic Coliphage Profiles of Produce and Environmental Samples from Farms in Northern M�xico. Food Environmental Virology. Sep;8(3):221-6.
- Type:
Journal Articles
Status:
Submitted
Year Published:
2016
Citation:
Newman KL, Jaykus LA, Leon JS. (2015) Norovirus transmission during produce harvest and packing: A quantitative microbial risk assessment modeling approach. Submitted to Journal of Food Protection.
- Type:
Journal Articles
Status:
Other
Year Published:
2016
Citation:
Morril V, Fabiszewski de Aceituno A, Bartz F, Heredia N, Garcia S, Shumaker DJ, Grubb JE, Arbogast JW, LEON J (2016). Visible Soil as an Indicator of Bacterial Concentration on Farmworkers Hands. In preparation
- Type:
Theses/Dissertations
Status:
Other
Year Published:
2016
Citation:
Marsh Z. The Development of a Quantitative Microbial Risk Assessment Model to Evaluate the Efficacy of Produce Rule Interventions to Reduce Norovirus and Hepatitis A Virus Contamination of Fresh Produce on Farms and Packing Facilities. Masters in Public Health Epidemiology Rollins School of Public Health
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Newman KL, Bartz FE, Johnston L, Moe CL, Jaykus LA, LEON JS. Microbial Loads of Fresh Produce and Packing Equipment Surfaces Collected Near the US and Mexico Border are Associated in Packing Facilities. International Association for Food Protection, St. Louis, MO. July 31, 2016.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Marsh Z, Talekar S, Bartz FE, Fabiszewski de Aceituno A, Jaykus LA, LEON JS (2016). A Method for Norovirus Detection in Agricultural Water, Produce, and Hand Rinse Samples. International Association for Food Protection, St. Louis, MO. July 31, 2016.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Morril V, Fabiszewski de Aceituno A, Bartz F, Heredia N, Garcia S, Shumaker DJ, Grubb JE, Arbogast JW, LEON J (2016). Visible Soil as an Indicator of Bacterial Concentration on Farmworkers Hands. International Association for Food Protection, St. Louis, MO. July 31, 2016.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Morril V, Fabiszewski de Aceituno A, Bartz F, Heredia N, Garcia S, Shumaker DJ, Grubb JE, Arbogast JW, LEON J (2016). Visible Soil as an Indicator of Bacterial Concentration on Farmworkers Hands. Latin Food 2016, Cancun, Mexico. November 9, 2016.
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Progress 02/01/15 to 01/31/16
Outputs
Target Audience:Target audiences: One target communities are the main producers of jalapeno, tomato (open field) and cantaloupe melon in the north-east of Mexico where the samples were collected. More broadly are Mexican and American produce growers, importers, distributors, retailers, governing bodies responsible for food safety regulations, and members of the agricultural and food safety research communities in both countries all stand to benefit from the knowledge generated by the project. We have also mentored 17 individuals in food safety ranging from staff, faculty, masters students and undergraduate students in the US. Efforts: We have specifically reached out to members of the produce community, including United Fresh, the Produce Marketing Association, hand hygiene industry, including GOJO Industries, USAID, and have participated in academic-government-industry meetings including the: International Association for Food Protection '15 meeting and the Western Center for Food Safety '15 meeting. Lastly, we presented our work formally to the Center for Applied Nutrition and Applied Nutrition, Food and Drug Administration in 2015. We have also submitted a publication to the Journal of Food Protection. We also provide annual trainings to thesis, laboratory, and research students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the current reporting period, a total of 17 individuals have received professional training in project management, experimental design, laboratory techniques, statistical analysis, outreach and communication, and scientific writing. Project staff included two undergraduate students, 9 graduate students, 3 technical staff, and 2 scientists (1 faculty) during the reporting period. One MPH student obtained a 25K NoroCORE fellowship on this work thanks to his preliminary work on this project. How have the results been disseminated to communities of interest?The products described above, including submitted peer reviewed publications and presentations at local and professional conferences have all served to disseminate research results and engage communities of interest. What do you plan to do during the next reporting period to accomplish the goals?Generally, we plan to continue work on all objectives as they are all advanced but in various stages of work: Objective 1: We plan to complete concentration and amplification of all the remaining samples. Once completed, we plan to begin an initial draft of the NoV and HAV concentration and optimization method for publication. Objective 2: We plan to complete our systematic review of indicators of NoV and HAV. We also plan to link our Objective 1 results to our existing database of bacterial and viral indicators for preliminary analysis. Objective 3: We plan to take our submitted manuscript through to publication and expand our model to HAV and data from Objective 1. Objective 4: We plan to collect stakeholder feedback on our preliminary model and Produce Rule scenarios and update our existing QMRA model to reflect these suggestions. We then plan to incorporate Objective 1 results and develop a draft publication for peer-review.
Impacts
What was accomplished under these goals?
Objective 1: We first developed and optimized a method for viral concentration and NoV (GI and GII) and HAV detection on produce and environmental samples which included a matrix inhibition control. This work was submitted to IAFP'16 for consideration as a poster presentation. We then will apply this two-part method (concentration and detection) to our collected produce and environmental samples. We began with 636 samples representing 85 production chains of tomato, cantaloupe, and jalapeno pepper from field production through off-farm distribution, including 7,860 units of produce, 1,026 farm worker hand rinses, 51 kg of soil, and 567 L of source and irrigation waters from ten participating farms in northern Mexico. Of these samples, we have concentrated 80% (325/404) samples: tomato, cantaloupe, jalapeno pepper, hand rinses, irrigation, and source water. We have also completed a first NoV GI and GII amplification on 50 of these samples and are in the process of repeating matrix inhibited samples. NoV (GI and GII) and HAV RNA are present in the produce environment but we require additional samples amplified before providing prevalence estimates. Objective 2: While we complete Objective 1 and compare our NoV and HAV laboratory results to our database of bacterial indicators (Coliforms, E. coli, Enterococcus, coliphage), we have begun a systematic review on PubMed to identify published indicators of NoV in the production environment. We began with 297 titles and after applying our inclusion and exclusion criteria, we have 67 articles that we reviewed and extracted data for our database. We have identified a variety of viral and bacterial indicators whose relationship has been analyzed against the presence of NoV on produce, soil, water, and hands. There is no clear consensus of an indicator that is consistently present when NoV is present. We are considering repeating this systematic review and broadening to other databases (e.g. Web of Science, Agricola) and to HAV. Objective 3. We have completed a preliminary QMRA model for NoV contamination on fresh produce in the production environment. This was presented at IAFP'15 and a publication has been submitted to the Journal of Food Protection. We used a stochastic QMRA model developed in R with data on key parameters obtained from the literature to test the impact of hygiene and worker health practices on NoV contamination of produce during harvest and packing. Based on this model, contamination was greatest for the first 5 to 10 items harvested and packed by infected workers after using the restroom, declining by over 2 log10 to approximately 20 virions/cm2 by the 10th item. Complete hand washing compliance by infected produce harvesters reduced contamination by 20.2% compared to the baseline model, and complete compliance by infected packers reduced contamination by 26.3%. In addition, increases in hand washing efficacy reduced contamination; a 2-fold improvement in efficacy reduced contamination by 29.9%. The model also showed that contamination of produce declined to below a detectable concentration (i.e., 10 virions/cm2) when infected workers were shedding less than 107 NoV virions/g stool, suggesting that workers may not require furlough until the end of all detectable NoV shedding. We are planning to extend this to HAV as well. Objective 4. A stochastic QMRA model for four produce groups (leafy greens, melons, berries, produce from vines/stalks) was developed with Crystal Ball using literature-based data of parameters such as: NoV in irrigation and rinse water, on worker hands, and on equipment. A sensitivity analysis identified the most influential parameters. A variety of scenarios were performed to determine baseline contamination risk and evaluate select Produce Rule guidelines (improved water quality, glove use, handwashing, equipment cleaning). Among the four produce groups, average baseline risk of NoV infection per produce item was lowest for berries (8.7 x 10-4/item [SD=5.27 x 10-4]) and highest for melons (9.0 x 10-2/item [SD=5.0 x 10-2]). Glove use and handwashing demonstrated the greatest risk reduction capacity (199% and 49%, respectively) compared to baseline risk. The sensitivity analysis also suggested that worker hand NoV concentration, across all QMRA model inputs, were most highly correlated with consumer risk of infection across all produce groups (r = 0.925 - 0.95). When all four Produce Rule guidelines were implemented, risk of infection was reduced between 337% (berries) and 409% (melon). We plan to incorporate data from Objectives 1-2 and share these preliminary results with stakeholders (CDC, FDA, produce industry groups) for feedback and refinement. One MPH student obtained a 25K NoroCORE fellowship on this work thanks to his preliminary work on this project. Dissemination and outreach activities included submitted publications and presentations listed under products to academic, industry, and government stakeholders. These deliverables and outputs will contribute to both research on produce safety and activities in U.S. agriculture to minimize viral hazards on fresh fruit and vegetables.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2015
Citation:
Newman KL, Jaykus LA, Leon JS. (2015) Norovirus transmission during produce harvest and packing: A quantitative microbial risk assessment modeling approach. Submitted to Journal of Food Protection.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2015
Citation:
Heredia N, Caballero C, C�rdenas C, Molina K, Garc�a R, Sol�s L, Burrowes V, Bartz FE, Fabiszewski de Aceituno A, Jaykus LA, Garc�a S, and Leon JS. Microbial indicator profiling of fresh produce and environmental samples from farms and packing facilities in northern Mexico. Submitted to Journal of Food Protection.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2015
Citation:
Bartz FE, Watson D, Heredia N, Fabiszewski de Aceituno A, Jaykus LA, Garcia S, Leon JS. Somatic coliphage profile of produce and environmental samples from farms in northern Mexico. Submitted to Food and Environmental Microbiology.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Juan Leon, Faith Bartz, Santos Garcia, Norma Heredia, Lee-Ann Jaykus. 2015. Clean Greens: Routes of Fresh Produce Bacterial and Viral Contamination and Hygiene Interventions among Farms and Packing Facilities on the U.S.-Mexico Border. Washington, D.C. Center for Food Safety and Applied Nutrition. Oral presentation.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Juan Leon, Faith Bartz, Anna Fabiszewski de Aceituno, Santos Garcia, Norma Heredia, Lee-Ann Jaykus. 2015. Clean Greens: Fresh Produce Contamination and Hygiene Interventions among Farms and Packing Facilities GOJO Industries. Akron, OH. Oral presentation.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Anna Fabiszewski de Aceituno, Kira Newman, Lee-Ann Jaykus, Juan Leon. 2015. Norovirus transmission on farms during produce harvest and packing: A quantitative microbial risk assessment model Efficacy of Alcohol-based and Soap-based Hand Hygiene Interventions on Farmworker Hands Soiled during Harvest. Annual meeting of the International Association for Food Protection. July 25-28 2015. Portland, Oregon. Oral presentation.
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