Joint Institute for Food Safety and Applied Nutrition
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1 Joint Institute for Food Safety and Applied Nutrition Influence of Pre-harvest Antibiotic Pesticide Treatment on the Native Microflora of Apple and Pear Blossoms, Leaves, Fruit, and Cider and its Implications for Food Safety Dr. Christopher S. Walsh
2 Collaborators Arthur Miller, CFSAN Brian Shawn Eblen, CFSAN Andrea Ottesen, UMD Doug Price, UMD Keedysville JIFSAN undergraduate students
3 Background Limited knowledge on relationship between unpasteurized apple cider safety and: Good agricultural practices Known -Avoid drops (2 log safety improvement) -Use U.S. cider grade apples or better (<1 log improvement) Good manufacturing practices Known -Contaminated lots
4 Unpasteurized Apple Cider as a Public Health Problem Cider-Associated Outbreaks 1974 Salmonella (300 cases) 1980 E. coli O157:H7 (23 HUS cases) 1993 Cryptosporidium (213 cases) 1996 E. coli O157:H7 (3 outbreaks) Cryptosporidium (213 cases) 1999 E. coli O157:H Cryptosporidium (>5 cases) 2004 E. coli, Cryptosporidium, Camplobacter
5 Fire Blight (Erwinia amylovora) Gram negative bacterial pathogen that attacks members of the Rosaceae Major problem in apple and pear orchards Lethal to many pear varieties Lethal to popular apple rootstocks Limits apple exports to Japan Treatments: Copper, Streptomycin and Oxytetracycline Streptomycin-resistant Erwinia epidemic in MI, 2001
6 Data Gaps: What is the effect of fireblight controls on microbial ecology in the orchard? Unknown : Influences of antibiotic use Effect on overall orchard microflora Effect on presence of Gram negative human pathogens responsible for illness outbreaks Organic pesticides?
7 Does less pesticide reduce chemical risks but increase biological risks? Plot 1: Unsprayed Transitional Organic Plot 3: Conventional Spray Schedule
8 Experimental Design: Plot 1: Unsprayed Transitionally organic Plot 2: Early Sprays Only Conventional sprays until June 15 th, Unsprayed July, Aug, Sept. Plot 3: Conventional Spray
9 Experimental Procedure: Duplicate sets of apples are collected from four locations on each tree in the three experimental plots. Wash is prepared from surface microflora and 20 ml of water and 0.1% peptone buffer ph 6.8 Dilutions of the wash are plated onto general and selective media. Populations are counted, recorded, and analyzed statistically using SAS.
10 Preliminary Experiment (2002) Dependent Variables Yeast and Molds, BHIA, MacConkey, Total Coliforms Independent Variables Pesticide Application Tissue (Leaf, Fruit and Shoot) Position within Tree Samples taken in August, Sept. and Oct.
11 Preliminary Results (2002) Significant Effects Pesticide Usage (Half log reduction with Pesticide Usage) Position within Tree Non-significant Effects Leaf and Fruit Counts Similar Shoot Counts Extremely Variable
12 Less-selective Media Brain Heart Infusion Agar (BHIA) Total aerobic count 3M Petri Film Yeasts and Molds
13 Selective media MacConkey (Gram negative bacteria) Petri Film Total Coliforms (30-37C)
14 Total Aerobic Count BHIA (2003) Log CFU / Apple (SE) Treatment Early-Season Late-Season Full Spray 4.38 (.06) 4.35 (.03) Half Spray 4.49 (.06) 4.39 (.02) No Spray 4.55 (.06) 4.59 (.03)
15 Selective Media for Gram Negative Bacteria MacConkey(2003) Log CFU / Apple (SE) Treatment Early-Season Late-Season Full Spray 3.30 (.03) 4.23 (.05) Half Spray 3.31 (.04) 4.57 (.05) No Spray 3.53 (.05) 4.81 (.05)
16 Selective Media 3M Petri Film Total Coliforms (2003) Log CFU / Apple (SE) Treatment Early-Season Late-Season Full Spray 1.93 (.03) 2.75 (.05) Half Spray 1.74 (.03) 3.06 (.05) No Spray 1.79 (.03) 3.27 (.05)
17 Laboratory Cider (2003) Plate Counts Log CFU / cm 2 Treatment BHIA MacConkey Total Coli Full Spray Half Spray No Spray
18 Discussion: Is the log reduction a significant GAP? How does it affect human pathogens? Option 1: Reduce risk by direct kill of human pathogens? Option 2: Increase risk for human pathogen survival if native microflora are no longer occupying competing niches?
19 Organic Production and GAPs. To be profitable, organic orchards require cider production. What are the implications on food safety caused by the change from conventional to organic production systems?
20 Wye Experimental Organic vs. IPM (Conventional) Orchard
21 AREAS OF RESEARCH INVESTIGATION JIFSAN FUNDING Food Safety Issues Fruit Quality AGROECOLOGY FUNDING Specialty Crop Potential Economics Insecticide, Herbicide, Fungicide and Fertilization Requirements and Recommendations
22 ORGANIC VS. CONVENTIONAL PLOTS Design: Randomized orchard blocks: 2.5 acres 50 feet between conventional and organic conditions (MDA) Organic Certification anticipated in fall, 2005
23 NINE CULTIVARS 3 Disease resistant apple cv LIBERTY, GOLDRUSH, ENTERPRISE LIBERTY OLYMPIC GALA 3 Asian pear cv OLYMPIC, ATAGO, NITAKA ENTERPRISE NITAKA FUJI 3 Commercial apple cv GALA, FUJI, CORTLAND GOLDRUSH ATAGO CORTLAND
24 Pressure treated wood for tree support in the conventional plots Strips prepared with herbicide, Gramoxone Fertilized w/calcium nitrate Aluminum poles for tree support In the organic plots Dedicated organic product sprayers used for all applications. Mowers cleaned before use in organic plots.
25 PLANTING YEAR: PEARS PLANTED 4/17/03 APPLES PLANTED 4/20/03 ORGANIC MATERIALS 2003 IPM ORCHARD MATERIALS /30/03 Fertilize Nitro Mix 6/6/03 Herbicide: All Down 6/10/03 Herbicide: 20% acetic acid 7/4/03 Insecticide: Pyganic 7/7/03 Herbicide: All Down 7/10/03 Fertilizer: Nitro Mix 7/13/03 Pyganic & Sulfur 7/25/03 Pyganic, Sulfur & Surround 9/16/03 Pyganic, Sulfur & Surround 3/24/03 Herbicide: Roundup 4/30/03 Fertilize Ca(NO3)2 5/29/03 Herbicide: Gramoxone 7/2/03 Herbicide: Roundup 7/4/03 Imidan, Captan, Provado 7/10/03 Calcium Nitrate 7/17/03 Captec & Provado 7/30/03 Captec, Sulfur, Imidan, Nova 8/19/03 Captec, Sulfur, Imidan, Nova 9/6/03 Captec, Provado, Topsin 9/16/03 Roundup
26 ORGANIC MATERIALS 2004 CONVENTIONAL MATERIALS /10/04 Pyganic, Sulfur, Surround 4/20/04 Pyganic, Sulfur 4/30/04 Sulfur 5/5/04 Sulfur, Pyganic & Surround 5/13/04 Sulfur, Pyganic & Surround 5/24/04 Sulfur, Pyganic & Surround 6/7/04 Pyganic, Sulfur, Surround, Entrust 6/17/04 Pyganic, Sulfur, Surround, Entrust 6/24/04 Fertilize McGeary /15/04 Pyganic, Sulfur, Surround, Entrust 7/22/04 Pyganic, Sulfur, Surround, Entrust cont. 4/10/04 Warrior, Dithane, Rubign 4/20/04 Solican & Geamoxone 4/30/04 Dithane & Topsin 5/5/04 Mancozeb & Imidan 5/13/04 Mancozeb, Imidan & Topsin 6/7/04 Imidan, Thiodon, Captec, Topsin 6/28/04 Imidan, Nova, Provado, Captec 7/15/04 Imidan, Nova, Provado, Captec 7/22/04 Imidan, Captec, Topsin, Vydate 8/4/04 Roundup 8/5/04 Imidan, Captec, Topsin, Thiodon 8/20/04 Captec, Topsin & Sevin
27
28 ORGANIC CONVENTIONAL AUGUST 25 TH HARVEST OF ATAGO PEARS-16 months after planting orchard.
29 Preliminary Greenhouse Study, 2004 Purpose- To develop a protocol for testing applications of coliform bacteria under controlled conditions. March. Plant 50 apple trees in container nursery in soilless mix. June. FSIS clearance sought to use GFP and K12 strains under controlled conditions.
30 Preliminary Greenhouse Study, 2004 July and August. Greenhouse background/safety tests. Swab for coliforms 4 separate occasions. Low background level found in greenhouse. Log 2-log 3 levels in floor drains. August. Move container grown trees into greenhouse and begin shoot bioassays.
31 FACTORIAL STUDY OF K12 SURVIVAL EFFECT OF ALUMINUM FOIL
32 Preliminary Greenhouse Study, 2004 September. Controlled inoculations using K12 suspended in solution with non-ionic surfactant allow uniform wetting, and controlled treatments. A factorial testing incubation temperature (22 vs 37) and humidity demonstrated that aluminum foil covers over beakers held at 22 C allowed recovery of inoculated K12 for 7 to 10 days.
33 Research to be completed in early-2005 Compare counts in cider made from Unsprayed and Sprayed trees at Keedysville orchard Compare bacterial counts in cider made from organic and conventional apples and pears from Wye Test the effects of organic pesticides survival of K12 and/or GFP on greenhouse-grown shoots in the laboratory
34
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