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WISCONSIN CRANBERRY BOARD, INC. 2010 PROGRESS REPORT SUMMARIES The following progress reports were provided to the Wisconsin Cranberry Board, Inc. by the individuals and groups that received project funding during the 2009-10 fiscal year. These reports were presented to the WCB at or prior to the Budget Meeting on March 25, 2011. Copies of the full reports can be obtained from the authors or from the Wisconsin Cranberry Board, Inc. office. Crop Research Cranberry Fruit Rot Control in New and Established Plantings Principal Investigator: Patricia McManus, UW Madison, Department of Plant Pathology Project Summary: Cranberry fruit rot in Wisconsin is caused by a complex of at least 8 distinct fungal species. This project aims to identify the major cranberry fruit rot pathogens in Wisconsin and determine which fungicides are effective in controlling specific fungi. Early results suggest that the broad-spectrum fungicide Bravo is more effective than other registered fruit rot fungicides (Abound, Dithane, Evito, Indar) for controlling Colletotrichum and Coleophoma, the most prevalent pathogens at research sites and in miscellaneous diagnoses in 2009 and 2010. Objectives: 1) Determine which fungicides are effective in controlling fruit rot pathogens, especially Colletotrichum spp. and Phyllosticta vaccinii (June-September). 2) Determine the effect of evaporative cooling on duration of leaf/fruit wetness, canopy temperature, and effect on early rot in newer plantings (June-September). 3) Monitor survival of Colletotrichum spp. and Phyllosticta vaccinii in water and at various temperatures (November-December). 4) Perform diagnoses of cranberry fruit rot problems and develop a manual/web site to assist in diagnoses (June-January). Revision of Objectives: Objectives 1 and 4 have been conducted according to plan, and progress is summarized below. Objectives 2 and 3 were reconsidered and deleted. In lieu of these deleted objectives, experiments aimed at determining the effect of fungicides applied after hail injury were conducted. Summary of Accomplishments: Objective 1. Determine which fungicides are effective in controlling fruit rot pathogens, especially Colletotrichum spp. and Phyllosticta vaccinii. This work is complementary to the fungicide efficacy trials done by Jack Perry. Two trials (Trial A and Trial B) were done in 3-year old GH1 beds where early rot has been a problem in recent years. A third trial (Trial C) was in an older planting of Stevens where Colletotrichum had been isolated at high levels in previous years. Trials A and B included treatments of Abound (azoxystrobin), Indar (fenbuconazole), Bravo (chlorothalonil), Dithane (mancozeb), and Evito (fluoxastrobin) applied at full bloom and again during early fruit set. In late September or early October, 20 rotten fruit from each of five replicate plots were collected, for a total of 100 berries per treatment at each site. Fungi were isolated on potato dextrose agar, and after several days, identified by colony characteristics and spore morphology. To keep the experiment manageable, fungicide specificity for registered fungicides only was tested. The incidence of fungal pathogens varied among sites, and among replicates of treatments at a given site. The predominance of Coleophoma (ripe rot) at all three sites was not expected. Although this is a common cranberry pathogen, it is not typically seen at such high levels. Bravo was the only fungicide that markedly reduced the incidence of Coleophoma. As expected, the incidence of Colletotrichum gloeosporioides and C. acutatum (both cause bitter rot) was high, especially in the established bed with a past history of bitter rot (Trial C). Bravo was the most effective fungicide in controlling C. gloeosporioides. Indar was not effective in controlling Colletotrichum, and other fungicides were intermediate. Physalospora vaccinii (blotch rot) was more prevalent in the newer plantings than the established site. The significance of this finding is not clear but will be considered in the future. Phyllosticta vaccinii (early rot) was nearly absent in rotten berries at the time of harvest. However, at the two early rot sites (Trials A and B), it was present in about 20-30% of rotten berries collected from untreated areas near the ditch in late August. It is likely that this slow-growing fungus was out-colonized by other fungi during September or that its incidence was generally lower in our untreated plots than along the ditch. 2010 Progress Report Summaries March 25, 2011 Page 2 of 14 Tentative conclusion: The broad-spectrum fungicide Bravo is more effective than other fungicides for controlling Colletotrichum and Coleophoma, the most prevalent pathogens at these sites in 2010. Objective 4: Perform diagnoses of cranberry fruit rot problems and develop a manual/web site to assist in diagnoses. In 2009 diagnoses were performed and followed up with recommendations. In 2010, 9 diagnoses for grower collaborators were performed. The Plant Disease Diagnostic Clinic performed 26 diagnoses, all of which were followed up with recommendations. In 2009, dying uprights, especially from younger beds, were the most common sample submitted, whereas in 2010 fruit rots were the greatest concern. Pathogenic fungi associated with the dying uprights were not found consistently in either year. Colletotrichum spp. (bitter rot) Coleophoma (ripe rot), and Physalospora (blotch rot) were the prevalent fruit rot pathogens in both years. Phyllosticta vaccinii (early rot) was not detected in 2009, but in 2010 leaf and/or fruit rot symptoms were detected in younger (3 years or younger) plantings of GH1 and Demoranville. Pesticide Screening for Cranberries Principal Investigator: Jed Colquhoun, UW Madison, Department of Horticulture Cooperators: Dan Mahr, UW Madison, Patricia McManus, UW Madison, Jack Perry, UW Madison Objectives: The mission of the 2010 program was to investigate fungicides, insecticides and herbicides for use in cranberry production. Objectives were to: 1) Investigate pesticides currently registered for use in cranberries to refine their use patterns and to further identify their pest control spectrum; and, 2) Investigate pesticides not currently registered for use in cranberries for their potential to address existing pest problems. Summary: In the 2010 growing season, thirty-three field trials were conducted on twenty three Wisconsin marshes including 14 insecticide trials with a varying number of treatments at each site evaluating treatments for 7 target pests and 9 fungicide trials on three marshes with 14 treatments conducted. Insecticide research focused on product evaluations for tipworm, fireworm, flea beetle, fruitworm loopers and white grub control, as well as a refinement of recently registered insecticides. Fungicide evaluations focused on potential products for fruit rot control. The purposes of the 2010 herbicide trials were to investigate new post-applied products for possible uses in cranberries; seek control solutions for problem weeds (escapes); investigate Callisto on new plantings on high pH soils and, continue to investigate herbicide candidates for dodder control. INSECTICIDE TRIALS Cranberry fruitworm, Sparganothis fruitworm, and black-headed fireworm are the primary insect pests in Wisconsin cranberries. Most acres are treated at least once per season for one or more of these pests. Tipworm, loopers/spanworms and flea beetles are secondary pests; in any given season some acres are treated for these pests. Cranberry weevil, cranberry girdler and white grubs are also occasional pests; there are no efficacious insecticides registered for the control of these pests. Fourteen insecticide trials were conducted in 2010: four for fruitworms, two for fireworms, two for loopers, two for tipworms, two for flea beetle, one for leafhoppers and one for white grubs. The number of treatments evaluated varied with the pest and trial site. Recently registered insecticides, Assail, Knack, Rimon, Delegate, Belay, Intrepid, several older standards, Imidan, diazinon, Orthene, Lorsban, and several non-registered products were evaluated both alone and in tankmix combinations. Since trial sites were selected based on existing or developing insect populations all trials had moderate to heavy testable pest pressures. All of the registered products performed much as expected. The older organo-phosphate products were broad-spectrum across most test pests and were generally efficacious as long as the pest was present at the time of the application. Control ranged from acceptable to excellent. The newly registered products, particularly the insect growth regulator-types (IGR), were more pest-type specific. Lepidopteron pests were controlled well, the dipterans less so and the other pests mostly not controlled by the IGRs. Although all of the IGR products were generally equally efficacious, the timing of applications with these products was critical to performance; late egg-to-early instar applications were efficacious whereas applications to later instars applications were significantly less effective - to be expected with these types of insecticides. Tank mixes of the newer products with the organophosphates lessened the necessity for precise timings of applications. Of the newly registered products Assail, Intrepid and Belay were efficacious. Although the currently registered rate of Belay (4 oz/a) provided good, but marginally acceptable control, the higher rates (6 and 8 oz/a) demonstrated improved efficacy. One of the candidate insecticides was a stellar product. 2010 Progress Report Summaries March 25, 2011 Page 3 of 14 HERBICIDE TRIALS New Post Product Trials In our 2009 and 2010 trials, three of the experimental products caused discernable crop response. For one, the crop response was detectable season long. For the other, crop response was less long lasting but still unacceptable. Although the crop responses induced by either product did not result in significant yield reduction, the visual responses were unacceptable. The visual crop response induced by a third product was minor, however several tested treatments of this product resulted in significant crop reductions. Two experimental products demonstrated good promise for use in Wisconsin cranberries. The weed control spectrum for both of these products would make these great companion products for Callisto as they provide good control of weeds that are weaknesses of Callisto. One of these products provided good control of St. Johnswort, dodder and yellow loosestrife and demonstrated activity on maples. No crop responses were noted with either of these products. Callisto on New Plantings on High pH Soils Four small plot trials were established on cranberries planted in 2009 on high pH soils. Four rates (1, 2, 4, 6 oz/a) of Callisto with and without non-ionic surfactant were evaluated. Each rate was applied in early May when day and night temperatures were low and a separate set of the same treatments was applied in late May when day/night temperatures were more moderate. Two of the trials were conducted on Stevens cranberries and two on Grygleski GH1 cranberries. Soils in the test sites had pH of 6.8, 7.3, 7.8, and 8.0. No significant Callisto-induced crop responses were noted in any of the treatments in any of the four locations. Herbicide rate, surfactant and cold stress did not affect cranberry response to Callisto. Soil pH correction measures were applied to the entire bed and by late season cranberry stand establishment was in process. Weed Escapes Callisto has been used extensively in commercial production for several years. Several weeds that are not controlled by Callisto are now creating problems. Those weeds are sweet vernal grass, creeping red fescus, cinquefoil, Solomon‟s plume, trees (maples, willows, popples, oaks) and dewberry. Sweet vernal grass. Large infestations of vernal grass are occurring infrequently in ever-enlarging patches in some beds. One trial was conducted. Select, Select Max and Poast provided excellent control of vernal grass. Very early applications, multiple applications (if allowed by the label), maximum allowed rates and the use of an appropriate surfactant were keys to successful control. Callisto and two candidate products did not control vernal grass. Although a glyphosate wipe provided moderate control of vernal grass, satisfactory coverage was difficult to achieve. Creeping red fescue. Creeping red fescue is mostly a problem in the bed margins of new plantings. This grass is often used to stabilize dikes and problem grass is likely coming in off of the dikes by erosion or miss-applications during seeding. Two trials were conducted. By the end of the 2010 season, none of the tested treatments appeared to be adequately controlling creeping red fescue; a spring evaluation will be more definitive. Tested products are Casoron, Evital, Select Max, Poast and Callisto. Cinquefoil, Solomon’s plume. Both of these weeds are becoming less problematic as Callisto is used more extensively. One trial on each of these weeds was conducted in 2010. The keys to the successful control of cinquefoil and plume are: 1. keep up a program of Casoron and Callisto early 2. use Callisto applied early (especially on plume), high rate, and with appropriate surfactant 3. expect it to take several years to achieve control Maples, Willows, Popples, Oaks. Two trials per tree type were conducted in 2010. Callisto will provide good control of willows, popples, and maybe oaks. The keys to good control with Callisto are high rates, use of a surfactant, and multiple applications beginning early in the season; mid-season applications are significantly less effective. Maples are more difficult to control. Callisto injures but does not kill them, as does one of the candidate herbicides. A combination of Callisto and the candidate product may prevent maples from successfully over-wintering. For all of these problem trees, a glyphosate wipe was efficacious though labor intensive. 2010 Progress Report Summaries March 25, 2011 Page 4 of 14 Dewberry. Two trials were conducted in 2010. None of the three non-registered products effectively controlled dewberry. Although two of these products had activity on dewberry they also had detrimental effects on the cranberries. The search for dewberry control continues. Dodder Trials In 2008 and 2009 there was success with a non-registered product for the control of dodder. This product is pending registration for uses in cranberries. In 2010 five trials were conducted in two marshes to investigate use patterns of the nonregistered product (rates, application timing, tank mixes) for dodder control. Three of these trials at City Point, WI had heavy dodder infestations and valid trials were conducted. The candidate product continued to be highly efficacious for dodder control; application timing of this product is critical to good control. Applications need to be made when the dodder strands first begin to appear in the cranberry canopy. It is likely that this timing coincides with dodder seed germination or just before the dodder vines abscise from the soil. Later applications inhibited dodder but did not prevent dodder vine matting. Callisto caused temporary chlorosis in the dodder but did not provide control. Combinations of the candidate product and Callisto did not provide enhanced control over comparable rates of either product alone. Two other candidate herbicides did not provided control of dodder. FUNGICIDE TRIALS In 2007-2009 late season fruit rots caused significant problems in Wisconsin cranberry production; in some marshes 30% of the harvested crop was lost to fruit rot. The 2010 growing season was conducive to fruit rot – warm with ample, periodic rainfall. Yield losses of 25-50% were experienced. This disease complex generally affects mature beds that are in full production. In 2009 isolated incidents of early rot were consequential problems; losses of 50 – 100% of the crop were experienced. In 2010 yield losses due to early rot were isolated but usually significant when they occurred. This disease and these losses generally occurred in 2-3 year old beds. Fruit Rot Three trials were conducted on two marshes that have experienced significant fruit rot problems in recent years. LeMunyon and Stevens were the subject varieties. Fourteen treatments were evaluated. Treatments included various timings of applications of the registered products Bravo, Abound, Evito and Indar. Three non-registered products were also included. Disease pressure was moderate-to-heavy in all three sites. Bravo and Abound were the most efficacious products. Indar and Evito were less effective. The current recommendation is for two applications of a fungicide at 50% bloom and at early post bloom. Additional applications at pre-bloom did not contribute significantly to enhanced disease control. There was a trend for improved disease control when later applications were made at late berry set. None of the three candidate fungicides was effective. Early Rot Two trials were conducted on two marshes that experienced significant early rot problems in 2009. Grygleski GH-1 was the subject variety. Ten fungicide treatments were evaluated. Treatments included two applications of the registered products Bravo, Abound, Evito, Dithane and Indar. Five non-registered products were also included. Early rot disease pressure was light-to-moderate in both sites. Bravo and Dithane provided mediocre disease control. The other products were ineffective. Breeding Cranberry for High Yields and Ease of Culture When Grown Under Wisconsin Conditions Principal Investigator: Brent H. McCown, Department of Horticulture, UW-Madison Cooperators: Wisconsin cranberry growers, Ocean Spray Cranberries, Inc. Co-Investigator: Eric Zeldin, Associate Scientist, Horticulture, UW – Madison Project Summary: The UW cranberry breeding program‟s first release, „HyRed‟, continues to be expanded and plantings in 2010 had good results with no problems observed or reported. The program‟s second release, „Sundance‟, is currently awaiting patent approval, but has been scaled up and commercial release should begin in 2012. Newer selections are under continued evaluation and scale up. Specialized precision planting with conventional vines should greatly accelerate the ability to rapidly scale up to full bed size for real world evaluations. Tetraploid selections were conventionally replanted to see if a dense uniform planting will result in improved fruit set and large berry size. Other activities not directly funded by WCB 2010 Progress Report Summaries March 25, 2011 Page 5 of 14 included a survey of bacterial associates with cranberry plantings and the use of composted cranberry leaf refuse as a biocontrol for other horticultural crops. 2010/11 Objectives: 1) Support and monitoring of „HyRed‟ growers and propagators. 2) Release of „A-X15‟ as a cultivar, expansion and preparation for commercial availability. 3) Continued evaluation and scale up of second generation selections. 4) Expansion of existing tetraploid plots using conventional propagation. 5) Develop model to study yield components with upright profiling. 6) Establish new breeding plots to examine crosses focused on exploiting expanded season. Summary of accomplishments: 1. Support and monitoring of „HyRed‟ growers and propagators. A significant number of new acres of „HyRed‟ were planted in 2010. There are now approximately 200 plus acres of „HyRed‟ in the ground, the vast majority in Wisconsin. A number of site visits were made to central and northern Wisconsin to view and assist growers with „HyRed‟ plantings, particularly new and second year plantings. 2010 was a good growing year and good establishment was obtained by the growers with no particular problems. Mowed beds were also observed. These beds demonstrated excellent regrowth, although some were overly lush mid-season, by the end of the season the vines had settled down and no problems were noted. 2. Release of “A-X15” as a cultivar, expansion and preparation for commercial availability. “WI92-A-X15”, also called „Sundance‟, was proven in 2009 to be worthy of release as a new cultivar based on yield, berry size, vigor, rebud and other traits. In 2010, grower/cooperators planted an additional fourteen and a half acres of „Sundance‟. The vines were vigorous and good establishment was observed. In cooperation with WARF, a plant patent application has been submitted to patent office and varietal protection is expected to be awarded within the next few months. Propagators have been secured and a number of acres will be available for mowing in 2012 for commercial release. While most of the „Sundance‟ was been mowed in 2010, one large un-mowed plot provided an example of resiliency in this cultivar. The plot was located in a bed of „Ben Lear‟ which was very lush, presumably due to the warm wet growing season in 2010. Random samples estimated the yield of the „Ben Lear‟ to be 174 B/a, while the „Sundance‟ was estimated at 373 B/a and a nearby „Stevens‟ bed at 271 B/a (typical for this area). The „Sundance‟ plot and „Stevens‟ bed were far less lush than the „Ben Lear‟. The large berry size of the „Sundance‟ was of particular interest. Size classing of the three cultivars revealed a higher percentage of yield of „Sundance‟ was in the larger berry size classes when compared to the other cultivars. 3. Continued evaluation of second generation selections. One second generation selection, “WI02-A4G-X1”, which has shown great promise for a number of traits, was scaled up to a four acre bed using a precision planting technique developed by a grower/cooperator (Cranberry Creek Cranberries). This allowed for rapid scale up with a vastly reduced amount vines. This planting yielded an excellent response and good establishment is expected. This process offers a significant advantage in accelerating the evaluation of new selections in full beds using convention vines. Six other second generation selections were conventionally scaled up to very large plots by grower/cooperators at two other sites for further evaluation. Ideally, these plots will be large enough for further full bed scale up using the precision planting technique if the desirable traits are maintained. Performance plots were evaluated at one site for yield and berry size by random samples with the majority of individuals displaying the traits for high yields. The evaluations will be repeated in 2011. 4. Expansion of existing tetraploid plots using conventional propagation. The existing plots of two selected tetraploids were mowed and the vines conventionally planted adjacent to the originals. The original plots were derived from transplants, which turned out to be a problem for tetraploids due to a lack of runnering and uneven establishment. This makes it hard to properly evaluate tetraploids as dense flowering and exclusion of diploid (normal) pollen sources is essential for good reproductive fertility. The conventionally planted vines had a good take in 2010 and even displayed some runnering. It is expected that these plantings will result in more uniform and densely established plots. If this results in dense flowering and good fruit set with at least five seeds per berry, then significant yield increase could be realized due to very large berry size. Flower bud 2010 Progress Report Summaries March 25, 2011 Page 6 of 14 set has always been good in the tetraploids, so it is likely dense flowering will occur by 2012. Fruit set and seed set will continue to be evaluated and if favorable, these plots will be expanded again in the future. Determining Nutrient Availability Profiles in Newly Planted and Established Cranberry Marshes Project Coordinator: Rebecca Harbut, Department of Horticulture, UW-Madison Cooperators: Western Ag Innovations, Inc., Cranberry Growers, Ag Source Labs Objectives: 1) Establish nutrient availability profiles for newly planted beds using currently established recommended rates of fertilizer applications. 2) Establish nutrient availability profiles for established marshes receiving current recommended rates of nutrient applications. 3) Determine the effect of pH on nutrient availability profiles. Progress Report: All data collection for this study is complete and is currently being analyzed so there is no data in the progress report, but will be shared when all analysis is complete. Project will continue to monitor root growth during 2011. When tubes were inserted into the soil there was significant disruption to root growth and therefore impacted the quality of the images obtained. After being in place for one year, the soil has settled around the tubes and will allow for higher quality images. Objectives 1 and 2: Nutrient Uptake/Availability. During the 2010 season, PRS (Plant Root Simulator) probes were placed in an established bed and a newly planted bed at 3 different locations. Probes were replaced every 2 weeks and sent to the lab for analysis. The last batch of results was recently returned and the data is being analyzed. Nutrient availability data is being compared to root growth periods and stage of growth to determine if availability correlates with key growth periods. Soil and tissue analysis were conducted on all beds included in the study. Fertilizer application records (rate, timing and form) will be analyzed and correlated to nutrient availability data. Root Growth. Each bed had 4 rhizotron tubes in the bed and images of root growth were taken approximately every 2 weeks. Efforts are underway to determine the most efficient method to quantify root growth to utilize this technology to monitor root growth. The tubes remain in the beds and we will continue to monitor growth in 2011. Objective 3: Locations were selected to provide 2 standard pH beds and one higher pH bed. The high pH bed was selected based on grower records and observations, however when soil analyses were conducted, the soil pH of all sites was very similar. The PI is currently working with the grower to determine the discrepancy. Iron in Cranberry Production: Sampling, Analysis and Effects on Plant Productivity Principal Investigator: Rebecca Harbut, UW Madison, Department of Horticulture Cooperators: Jayne Sojka, Ladybug IPM, Cranberry Growers, AgSource Labs, Beth Workmaster, UW Madison, Sarah Seibach, Department of Horticulture, UW-Madison Objectives: 1) Develop sampling and analysis protocol for cranberry tissue iron content. 2) Determine relationship between micronutrient soil content and tissue analysis. 3) Develop understanding of the relationship between plant iron status and plant productivity (yield, plant growth, photosynthetic capacity). 4) Evaluate effects of irrigation water on plant nutrient status. Summary of Accomplishments: Data collection for this project started during the 2010 growing season and has continued during the winter. Sarah Seibach, a Master‟s degree student, joined the Harbut lab in Sept. 2010 and has been working on this project since her arrival. She has been conducting a greenhouse study during the winter and will continue collecting field data through the 2011 season. Sarah has also been working with Beth Workmaster on the N uptake study funded by the WCB in 2010. Objective 1: Develop sampling and analysis protocol for cranberry tissue iron content. 2010 Progress Report Summaries March 25, 2011 Page 7 of 14 In the fall of 2010, tissue, soil and water samples were collected from 4 beds, 2 that are irrigated with high Fe content water and 2 with „average‟ Fe content in water. Six different washes were compared with current standard practice of not washing tissue. Effect of wash treatments on Fe and other nutrient contents were evaluated. In the high Fe beds, there was a significant difference in the Fe analysis reported with certain wash treatments. The pre-wash treatments did not affect the nutrient analysis of any other nutrients. Extension bulletin is being published to share the data. Objective 2: Determine relationship between micronutrient soil content and tissue analysis. Soil and tissue samples were taken from the same beds in the fall and analysis were compared to determine relationships between soil and tissue values. Data collected in 2011 will be added to the data set before all analysis is complete. As expected, there is little correlation between soil and tissue data for most nutrients. Data will be included in the above mentioned extension bulletin. Objective 3: Develop understanding of the relationship between plant iron status and plant productivity (yield, plant growth, photosynthetic capacity). In the spring and fall of 2010, preliminary productivity data were collected. Data collection will continue through 2011 with more intense data collection to determine yield parameters (uprights, flowers, fruit set) and physiological processes (photosynthesis) of the high and low Fe sites. Objective 4: Evaluate effects of irrigation water on plant nutrient status. A study to determine the impact of high Fe irrigation water is currently being carried out in the greenhouse. Cuttings collected in the fall were rooted and are being grown in sand culture. Irrigation water with different Fe content is being applied to the soil or to foliage to isolate the effect of Fe applied to soil vs. tissue. Nitrogen Response of High Yielding Cranberry Cultivars Project Coordinator: Rebecca Harbut, Department of Horticulture, UW-Madison Objective: Evaluation of Nitrogen Use Efficiency of standard cultivars compared to new varieties. Justification and Significance: Nitrogen (N) is a limiting nutrient in the cranberry production system and applications are regularly made to meet the demands of the plant. Preliminary work in the field indicate that new cultivars developed by the UW-Madison breeding program may have higher nitrogen requirements compared to older cultivars that have been industry standards. Understanding the nitrogen response of these new cultivars will assist in the future development of nutrient recommendations. The objective of this study is to generate preliminary data to compare nitrogen responses of these new high yielding cranberry cultivars to the current industry standards. Progress Report: Plant material was collected in the fall 2009 and cuttings were given their chilling period before setting up the greenhouse hydroponic system at the Arlington Agricultural Research Station Greenhouses. Cuttings were rooted and then transferred to the hydroponic system where N treatments were applied. After 3 days of treatment, there was a malfunction in the greenhouse over the weekend and temperatures rose to over 100oF and most of the uprights were severely stressed and did not recover. The project was terminated. New cuttings were collected in fall 2010 and given a chilling treatment. Cuttings have been rooted and are currently growing in a hydroponic system at the Walnut Street Greenhouse facility on the UW-Madison campus. This facility has much better control systems and plants have been growing very well. Treatments were started on March 1st and will continue for 4 weeks. After 4 weeks, the plants will be given a pulse of radio-labeled N to determine the uptake rates of the different cultivars. The experiment is expected to be completed in April. Annual Projects Wisconsin Cranberry Crop Management Newsletter – Volume XXIII Project Coordinator: Matt Lippert, Wood County UW Extension Agriculture Agent Cooperators: University faculty and staff, private cranberry consultants, Ocean Spray Cranberries, Inc., Cliffstar Corp., Clement Pappas & Co., Decas Cranberry Sales 2010 Progress Report Summaries March 25, 2011 Page 8 of 14 Summary: Ten issues of the CCM Newsletter were published between May and September of 2010. Copies were sent at no charge to managers of all known cranberry marshes in the state. The newsletter was also made available via the cranberry e-mail list. Wisconsin Cranberry School - 2011 Project Coordinators: Wisconsin Cranberry Research and Education Foundation; WSCGA Education Committee; Rebecca Harbut, UW Extension; Matt Lippert, Wood County UW Extension Agent. Objective: To conduct a 2 day grower educational program for all Wisconsin cranberry growers focusing on improved farm management and business practices. Summary: The WSCGA Education Committee met with UW Extension Faculty to evaluate previous schools and identify topics and speakers for 2011. The Wisconsin Cranberry School was held January 25-26 at the Stevens Point Holiday Inn Hotel and Convention Center. During the two day session topics relating to all aspects of cranberry production were presented with Peter Oudemans of Rutgers University as the featured speaker. Packets with relevant information were distributed to all attendees. Proceedings from the School will be available online and will be sent as hard copy to participants who request them. The 2011 Wisconsin Cranberry School attracted over 450 growers and industry people. Cranberry Weather Forecasts Project Coordinator: Wisconsin State Cranberry Growers Association Objective: To provide Wisconsin cranberry growers with accurate, regional weather forecasts. Summary: The WSCGA worked with a private weather forecasting consultant to develop regional cranberry weather forecasts. These forecasts were available to growers via a toll-free number and online at the WSCGA website www.wiscran.org . Service was available from April 15 through October 31. Brochure Printing Project Coordinator: Wisconsin State Cranberry Growers Association Objective: Provide members of the general public with information on cranberries through high-quality, professionally produced brochures. Summary: The grant was used to print and distribute 18,000 copies of the 2010 Fall Harvest brochure, printing of 15,900 copies of the recipe brochure and 8,750 copies of the Wisconsin Cranberry Activity Book. Product Sample Products Project Coordinator: Wisconsin State Cranberry Growers Association Objective: Provide promotional samples of cranberry products for distribution. Summary: WSCGA worked with cranberry handlers to secure sweetened dried cranberries which were then packaged into sample packets by Urban Processing in Wisconsin Rapids. Communications Program Project Coordinator: Wisconsin State Cranberry Growers Association Media Outreach & Harvest Promotion Objectives: (1) To provide targeted media with information on cranberries, cranberry products and information on the results of health related research on cranberry consumption. (2) Conduct fall harvest media campaign to educate the consuming public on the cranberry industry in Wisconsin. Summary: The harvest communications program entailed working with a public relations firm to develop key messages and themes, strategies, a plan and execution of the planned activities. The activities included a news release in conjunction with the crop projection announcement by USDA; an extended media outreach to target national and regional media including Associated Press; Cooperative effort with CMC to publicize tour by international trade representatives attending their annual marketing conference. a general use of the website to communicate with media and track efforts and visits. Individual components included working with the Rachel Ray television program to be featured as snack of the day assisting with the production of Curiosity Quest, a PBS children‟s program form California The 2009 earned media efforts generated about $1.3 million in advertising value and more than $3.89 million in publicity value. Traceable media placements of more than 568 had 23.3 million impressions. Additionally, these are conservative figures as some of the stories generated are not included in the value report, as the information was not available. 2010 Progress Report Summaries March 25, 2011 Page 9 of 14 Wisconsin State Fair Promotion Program Objectives: (1) Provide information on cranberries and cranberry growing to visitors to the Wisconsin State Fair. (2) Promote consumption and sales of cranberry products at the Wisconsin State Fair. (3) Increase overall awareness of cranberries and their economic, environmental and cultural importance to the state. (4) Educate the public on the health benefits of cranberry consumption. (5) Educate the public on the many cranberry products available and their uses. Summary: WSCGA contracted with the Wisconsin State Fair Park for space in the Wisconsin Products Pavilion. The booth space (10‟x30‟) was divided into two components: the first being an educational display, the second a sales area for cranberry products. The grant was used for promotion activities at the Fair including media drops of products, interviews on air, cranberry cooking demonstrations in cooperation with WE Energies, sponsorship of a State Fair cranberry Baking Recipe Contest and appearance by the cranberry mascot daily at the Fair. Milwaukee Brewer Radio Network Sponsorship Cooperators: Milwaukee Brewer Radio Network Objectives: (1) Conduct a paid advertising campaign to communicate health, environmental, tradition and economic messages on a regional basis. (2) Establish relationship between healthy sporting activities and cranberries. (3) Link cranberry growing tradition with other major state traditions. (4) Improve image of industry throughout the state. Summary: Wisconsin‟s cranberry growers were again sponsors of an in game feature of each Milwaukee Brewer Baseball Radio broadcast on the statewide network. The feature “On Your Plate”, the introduction of the umpires for each game was presented by Wisconsin‟s cranberry growers. The promotion also featured in game and post game mentions and Cranberry Night at Miller Park promotion. UW-Badger Sports Sponsorship Cooperators: Badger Sports Properties Objectives: 1) Conduct a paid advertising campaign to communicate health, environmental, tradition and economic benefits of cranberry growing on a regional basis. 2) Establish a relationship between health sporting activities and cranberries. 3) Link cranberry growing tradition with other state institutions and traditions. 4).Improve the image if the cranberry industry throughout the state. Summary: Wisconsin‟s cranberry growers were sponsors of the radio broadcasts of UW Madison Badger football, basketball and hockey from September 2010 through March 2011. In addition Bo Ryan, Men‟s basketball coach at UW Madison, served as the industry spokesperson. The commercials contained messages on environmental stewardship, health, tradition and economic importance of the industry. Wisconsin Cranberry Discovery Center–Operations & Promotion - Cranberry Museum, Inc. Project Coordinator: Barbara Hendricks, Director, Wisconsin Cranberry Discovery Center Objectives: Educate the public about Wisconsin‟s role as the nation‟s leading producer of cranberries. Summary: The Cranberry Museum, Incorporated utilized the grant to support activities at the Wisconsin Cranberry Discovery Center in Warrens, Wisconsin. The funds were allocated toward general operations and promotion efforts. Over 18,000 visitors came to the Discovery Center in the 2010 season. These numbers are based on customer counts from the POS system at the Center and hand counts by employees. They do not include traffic during the Warrens Cranberry Festival. Special promotions were supported with the funding including a Cranberry Blossom Day, Public Harvest Tours, leisure tours, regular news releases and day to day activities at the Center. Wisconsin Cranberry Discovery Center-Wetherby Cranberry Library Project – Cranberry Museum, Inc. Project Coordinator: Barbara Hendricks, Director, Wisconsin Cranberry Discovery Center Objectives: Organize and catalog new and existing materials at the Wetherby Cranberry Library. Summary: Since the hiring of a librarian to implement the project in May 2008, we have cataloged the existing materials at the Wisconsin Cranberry Discovery Center, a process that involved creating nearly 12,000 individual records; created the Wetherby Cranberry Library Digital Collection, the largest on-line repository of cranberry related items on the Internet. 2010 Progress Report Summaries March 25, 2011 Page 10 of 14 The collection features 400 unique objects, consisting of 2,500 .jpg images focusing on the history and importance of cranberry production in Wisconsin; established relationships with the Wisconsin State Historical Society, University of Wisconsin-Madison, Wisconsin Library Services and Wisconsin Heritage On-Line in order to further promote the importance of cranberry production in Wisconsin; integrated the Wetherby Cranberry Library Digital Collection into the Wisconsin Cranberry Discovery Center‟s Web site, thereby boosting the targeted audience‟s awareness of the Center and also making the collection more assessable and implemented both digital and archival protection strategies for the existing physical and digital collections. Since the middle of June through the middle of July, 2576 new items of the Edward Grygleski Donation have been cataloged. Of those items, 261 have been scanned. Ultimately, the entire collection will be scanned, optimized, and uploaded to the website for online display and search ability. There are currently 1007 items available on the Wetherby Cranberry Library Digital Collection. There are currently 250 objects (or materials composed of multiple pages) made up of 2576 items (an item is one side of a page) in the Edward Grygleski donation. Of these 2576 items, 355 items have been scanned, and re-sized for online display. 105 of the 355 digitized items have corresponding text files and are ready to be uploaded to Content DM. The library grant has allowed the library to purchase a computer workstation, necessary for supporting the continued cataloging, scanning, and uploading of new items to the digital resource center online. WISCONSIN CRANBERRY BOARD, INC. – CRANBERRY INSTITUTE HEALTH RELATED RESEARCH - PROJECT REPORT SUMMARIES The Wisconsin Cranberry Board, Inc. and Cranberry Institute have partnered to fund a variety of research projects related to the health benefits of cranberry consumption. Under this partnership the organizations issue a joint request for proposals. Researchers submit applications to the Cranberry Institute. The applications are reviewed by a Health Advisory Committee of the Cranberry Institute which makes recommendations for projects deserving funds. Those projects deemed worthy of funding are then jointly funded by Wisconsin Cranberry Board, Inc., the Cranberry Institute and other funding organizations. The Cranberry Institute then manages the research projects. In 2010 projects totaling $67,002 were funded by the WCB. The following are summaries of the reports of the projects funded by the partnership in 2010. Cranberry Polyphenols and Gut Health: Integrated Cranberry Health Research at UW-Madison Project Investigator: Jess Reed, UW Madison Objectives: Develop integrated cranberry health research program at UW Madison. Implement a pilot research project to develop novel mouse models to investigate cranberry PACS interaction with the gut mucosa and gut associated lymphoid tissue (GALT). Summary: Ten proposals have been submitted to funding entities with 3 funded and two pending. Two undergraduate research awards were considered, on funded and one pending. To date $184,000 in research dollars have been generated. New research directives involving UW Scientists include: Cranberry PAC fungal agents, Cranberry PAC biofilm and anti bacterial action, Effect of cranberry tannins on bacterial growth and cranberry products in meat processing. Significant progress has also been made in the development of a mouse model of GALT function. Tissue preparation protocols and fluorescent micropsy imaging parameters have been established. The collaborative network has focused on the ability of cranberry PAC to modulate pathogenic bacterial populations, virulence and invasively properties in the gastro-intestinal tracts and impact diseases such as UTI. Double-Blind Placebo Controlled Long-Term Study Of The Efficacy Of Cranberry Juice Consumption On Prostate Health Principal Investigator: Joe A. Vinson, PhD, Professor Chemistry Department, University of Scranton Co-Investigator: Donald L. Preate, MD, DeltaMedix, PC Joe has advised us that the cranberry beverage to be used was not available, due in part to the fact that it only has a nine month shelf life. Since the study will last six months, and the use of any expired product would be inappropriate, timing, and recruitment of the 40 subjects, is key. He has advised that the product will be available next month, at which time the clinical trial will begin. 2010 Progress Report Summaries March 25, 2011 Page 11 of 14 The Influence Of Cranberry Juice On Micro RNA Response In Senior Citizens. Principal Investigator: Carlo Selmi MD PhD, Assistant Professor of Medicine, Division of Rheumatology, Allergy and Clinical Immunology, Genome and Biomedical Sciences Facility, University of California at Davis This is also a clinical trial utilizing a cranberry beverage, and there have been delays in initiation due in part to product receipt. This issue is being addressed. ADDITIONAL HEALTH RESEARCH PROJECT UPDATES NathalieTufenkji. - Action of Cranberry Derived Proanthocyanidins against Biofilm Formation and Cell Infection: Potential for Protection against Urinary Tract Infection, Prostatitis, and Pyelonephritis. , Dept of Chemical Engineering, McGill University, Montreal, Quebec H3A 2B2, Canada. Primary areas of focus have included the following: Influence of PAC on Biofilm Formation. One of the early steps of structured biofilm development is the migration of bacteria by mechanisms termed swarming and swimming motility. Motility experiments conducted in our laboratory indicate that in the uropathogenic Escherichia coli CFT073, swimming and swarming motility are impaired in the presence of cranberry extracts (manuscript in preparation). We have also demonstrated that cranberry proanthocyanidins (PACs) and other tannin-containing materials can block swarming motility in Pseudomonas aeruginosa. Moreover, isolated patches of such materials can cause migrating bacteria to change their direction and move around the tannin source (manuscript in press). Mammalian Cell Infection. We found that cranberry PACs disrupt pedestal formation by enteropathogenic bacteria on epithelial cells, which is the first step required for bacterial infection. We also discovered that cranberry PACs can lead to cytoskeletal rearrangement of epithelial cells, which is another likely mechanism by which PACs inhibit bacterial infection. Experiments conducted in our laboratory further demonstrate that cranberry PACs inhibit invasion of HeLa cells by Salmonella. These findings are included in a manuscript currently being prepared for submission. List of New Presentations and Publications: 1. Hidalgo, G.; Ponton, A.; Fatisson, J.; O‟May, C..; Asadishad, B.; Schinner, T.; Tufenkji, N. “Induction of a State of Iron Limitation in Uropathogenic Escherichia coli CFT073 as Revealed by Microarray Analysis” Applied and Environmental Microbiology, 2011, 77, 1532-1535. 2. O‟May, C. and Tufenkji, N. “Pseudomonas aeruginosa Swarming Motility is Blocked by Cranberry Proanthocyanidins and Other Tannin-Containing Materials” Applied and Environmental Microbiology, 2011, in press. 3. Tufenkji, N.; Hidalgo, G.; Asadishad, B.; Ponton, A.; O‟May, C; Fatisson, J.; Schinner, T. “Influence of cranberry derived proanthocyanidins on the metabolism of a uropathogen: effects on cell motility, iron regulation and biofilm formation” presented at PACIFICHEM 2010, Honolulu, Hawaii, December 18, 2010. Cathy Neto - Cranberry Phytochemicals And The Role Of Hedgehog Signaling In Prostate Cancer Co-Principal Investigators: Dr. Catherine Neto Department of Chemistry and Biochemistry University of MassachusettsDartmouth and Dr. Robert Hurta, Department of Biology, University of Prince Edward Island Update: Administrative issues in both campus research offices delayed the availability of funds, which required coordination of external and internal grants and a subcontract to get started. Cranberry extract and fractions have been prepared and characterized for the allergy project by my student, and experiments in allergy cells are now underway in Rob's lab. The prostate cancer hedgehog signaling pathways experiments will begin this summer in Rob's lab, with final analysis of active fractions to be performed in my lab in the fall. Terry Camesano - Department of Chemical Engineering, Worcester Polytechnic Institute. Abstract of completed project for publication: A double-blind, placebo-controlled pilot clinical trial on the effect of cranberry juice cocktail (CJC) consumption on biofilm formation was conducted in 10 healthy women between the ages of 18 and 27. A single dose of 16 oz. of CJC or a placebo beverage was given to the volunteers, and urine samples were collected in the following 48 hours. Bacteria (E. coli B37, CFT073, BF1023, HB101, and S. aureus ATCC43866) were cultured in the urine samples supplemented with media and the amount of biofilm formed was measured using a crystal violet absorbance assay in a 96-well plate. In the urine of volunteers who had consumed CJC, biofilm formation was inhibited within 24 hours after CJC consumption, and biofilm 2010 Progress Report Summaries March 25, 2011 Page 12 of 14 formation started to increase after 48 hours by 49-67%. S. aureus showed the least biofilm formation after incubation with post-CJC urine. While E. coli CFT073 is a commonly studied uropathogenic E. coli strain and it expresses P fimbriae, E. coli B37 and BF1023 are antibiotic resistant and do not contain P fimbriae. This is the first study to show that biofilm formation in antibiotic resistant E. coli and a wide range of uropathogens, including S. aureus, can be prevented in the urine of healthy women who have consumed CJC. We also demonstrated the duration of the benefit, noting that antibiofilm activity in the urine peaks between 24 and 48 hours after drinking CJC, depending on the bacterial strain studied. Cathy Neto, - University of Massachusetts-Dartmouth. Her current updates include: We have a new publication in press on the anticancer/antiinflammatory compound ursolic acid in cranberries, cranberry products and other Vaccinium fruits. This study was made possible in part through grants from Cranberry Institute/Wisconsin Cranberry Board, so we have acknowledged your support in the article: Kondo, M, MacKinnon, SL, Craft, CC, Matchett, MD, Hurta, RAR, and Neto, CC. Ursolic Acid And Its Esters: Occurrence In Cranberries And Other Vaccinium Fruit And Effects On Matrix Metalloproteinase Activity In DU145 Prostate Tumor Cells. Journal of the Science of Food and Agriculture, available online (wileyonlinelibrary.com) DOI 10.1002/jsfa.4330). I wrote a chapter on ursolic acid derivatives and their occurrence in berries, which was just published in a book edited by Navindra Seeram and Gary Stoner: Catherine C. Neto, Chapter 2: "Ursolic Acid and Other Pentacyclic Triterpenoids: Anticancer Activities and Occurrence in Berries," in Berries and Cancer Prevention, Gary Stoner & Navindra Seeram (Editors), Springer, (2011). Rob Hurta at UPEI, his students, and members of my research group co-authored two new publications on cranberries and prostate cancer mechanisms: Déziel BA, Patel K, Neto C, Gottschall-Pass K, Hurta RA. Proanthocyanidins From The American Cranberry (Vaccinium Macrocarpon) Inhibit Matrix Metalloproteinase-2 And Matrix Metalloproteinase-9 Activity In Human Prostate Cancer Cells Via Alterations In Multiple Cellular Signalling Pathways.J Cell Biochem. 2010;111(3):742-54 and, MacLean, MA; Scott, BE; Deziel, BA; Liberty, AM; Nunnelley, M; Gottschall-Pass, KT; Neto, CC; Hurta, RAR. North American Cranberry (Vaccinium Macrocarpon) Stimulates Apoptotic Pathways In DU145 Human Prostate Cancer Cells In Vitro. Nutrition and Cancer, 63(1): 109-120 (2011). Also, I was invited to speak on cranberries and their effects on human health at the ACS National Meeting in Boston last summer (Aug. 2010), and I've just submitted a paper on cranberries for their symposium series volume, to be published by ACS Books: Christine A. Dao, Kunal D. Patel and Catherine C. Neto, "Phytochemicals from the Fruit and Foliage of Cranberry (Vaccinium macrocarpon) - Potential Benefits for Human Health." In addition, I submitted a pre-proposal in January to American Institute for Cancer Research for a collaborative research study on cranberries and breast cancer (Neto is PI; Hurta, Liberty and Ferreira, co-investigators). We were just notified last week that we were selected to submit a full proposal to AICR in May. The proposed study will look at cranberry's antibreast cancer effects in vitro and in two in vivo models (zebrafish and mouse). And finally, in December, a group of us submitted a R01 proposal to NCCAM, for a project investigating the mechanisms of action of cranberry's antibacterial activity. Terri Camesano is the PI and Joe Vinson and I are co-PIs on that proposal. Steven M. Lipson, Ph.D., (SVCP) - Professor, Department of Biology, St. Francis College It gives me great pleasure to report our accomplishments and new findings on the effect of cranberry juice and cranberry proanthocyanidins (PACs) on viral (rotavirus) infectivity. Firstly, I wish to report that we were recently informed that our more recent work was accepted for publication in Food and Environmental Virology. Our report shows in part, a protective effect of cranberry juice of the structural integrity of cell culture tight junctions as well as a protective effect on the physiology of host cells followed by rotavirus infection. Electron microscopy additionally, shows an entrapment of virus within PAC particles, and an envelopment of virus by solubilized PACs. These findings are significant, as they do not simply report input/output experiments, but show mechanisms related to juice and PAC antiviral activity. I will send you reprints of our article when available. We are greatly appreciative of the Cranberry Institute and members (e.g., Wisconsin Cranberry Growers‟ Association) for support over the years. Our research in this field could not have been performed without your generous funding. Presentations and publications: 1. Lipson, S.M. 2010. Invited speaker. Cranberry and grape juices reduce rotavirus infectivity in cell culture, and viral protein capsid integrity in cell-free suspension. 1st Annual Conference of the American Council for Medically Active Plants. July 20-23, 2010, Cook Campus Center, Rutgers, The State University of New Jersey. New Brunswick, NJ. Hosted by the Center ACMAP and the New Use Agricultural and Natural Plant Products Program (NUANPP, Rutgers University). 2010 Progress Report Summaries March 25, 2011 Page 13 of 14 2. Ozen, F. S., J. Nuamah, S. Mahaud, R. Gordon, and S. M. Lipson. 2010. Comestible juices and their secondary plant metabolites (e.g., proanthocyanidins) reduce rotavirus-associated dysfunction of monolayer tight junctions and compromise rotavirus capsid integrity in cell-free suspension. 43rd Annual MACUB Conference. Conference held at Molloy College, Rockville Centre, NY. Oct. 23rd, 2010. 3. Lipson, S.M., R.E. Gordon, F. S. Ozen, L. Karthekeyan, G. Stotzky. 2011. Effect of Cranberry and grape juices on tight junction function and structural integrity among rotavirus-infected money kidney epithelial cell culture monolayers. Food and Environ. Virol. (In press). 4. Ozen, F. S., S. M. Lipson*, D. Trotman, R. E. Gordon, S. Malmoud, and L. Karthikeyan. 2011. Investigations into the mechanism(s) of plant metabolites and comestible juices on the loss of rotavirus infectivity. Abstr., 111th Gen. Mtg., Amer. Soc. Microbiol. May 21-24, 2011. New Orleans. LA. (accepted for presentation). Jeg Babu. Dept. of Bioscience Research, College of Dentistry, U. of TN Health Science Center, Memphis. Attached please find two abstracts which have been accepted for publication in the Journal of Dental Research for presentation at the Annual IADR meeting, to be held in San Diego, Ca, March 16th to March 19th, 2011: Effects of Cranberry Components on Human Aggressive Periodontitis Gingival Fibroblasts. D. Tipton*, J. Babu, M. Dabbous. Dept. of Bioscience Research, College of Dentistry, U. of TN Health Science Center, Memphis Aggressive periodontitis (AP) causes rapid periodontal breakdown involving AP gingival fibroblast production of cytokines [i.e. interleukin (IL)-6, a bone metabolism regulator], and matrix metalloproteinase (MMP)-3. Lipopolysaccharide (LPS) upregulates fibroblast IL-6 and MMP-3, via transcription factors (i.e. NF- B). Cranberry (Vaccinium macrocarpon) inhibits LPS-stimulated macrophage and normal gingival (GN) fibroblast activities. Little is known of its effects on AP fibroblasts. Objectives: using AP fibroblasts, determine cytotoxicity of cranberry components or periodontopathogen (Fusobacterium nucleatum, Porphyromonas gingivalis) LPS ± cranberry components, and effects on NF- B activation and IL-6 and MMP-3 production. Methods: AP fibroblasts were incubated ≤6 d with NDM (high molecular weight non-dialyzable material derived from cranberry juice; 1-500 µg/ml) or LPS (1 µg/ml) ± NDM. Membrane damage and viability were assessed by enzyme activity released into cell supernatants and activity of a mitochondrial enzyme, respectively. IL-6 and MMP-3 were measured by ELISA. NF- B p65 was measured via binding to an oligonucleotide containing the NF- B consensus site. Data were analyzed using ANOVA and Scheffe‟s F procedure for post hoc comparisons. Results: Short-term exposure to NDM, or LPS ± NDM caused no membrane damage. NDM (≤ 100 µg/ml) or LPS ± NDM had no effect on viability ≤ 7d exposure. NDM (50 µg/ml) inhibited LPS-stimulated p65 (p≤0.003) and constitutive or LPS-stimulated MMP-3 (p≤0.02). NDM increased AP fibroblast constitutive or LPS-stimulated IL-6 (p≤0.0001), but inhibited GN fibroblast IL-6 (p≤0.04). Conclusion: Lack of toxicity of low NDM concentrations to AP fibroblasts in vitro, and its inhibition of NF- B and MMP-3, suggest that cranberry components may regulate oral fibroblast inflammatory responses and may be useful in prevention/treatment of gingivitis/periodontitis. Differences in NDM effects on AP and GN fibroblast production of IL-6 (which can have both positive and negative effects on bone metabolism) may reflect phenotypic differences and differing roles in bone metabolism. (Supported by The Cranberry Institute). Effects of Cranberry Components on Human Gingival Epithelial Cells. N. Zacharia*, D. Tipton, J.P. Babu, S. Rawal, and M. Dabbous. Depts. of Bioscience Research and Periodontology, College of Dentistry, U. of TN Health Science Center, Memphis. Gingival epithelial cells participate in periodontal inflammation and destruction, producing IL-6, which regulates osteoclastic bone resorption, and PGE2, which stimulates IL-6 production, osteoclastogenesis, and attachment loss. Cranberry (Vaccinium macrocarpon) components inhibit pro-inflammatory activity of LPS-stimulated human macrophages and normal gingival fibroblasts, but little is known of its effects on human gingival epithelial cells. Objectives: determine cytotoxic effects of cranberry components ± periodontopathogen (Fusobacterium nucleatum and Porphyromonas gingivalis) LPS on human gingival epithelial cells, and their production of IL-6 and PGE2. Methods: Smulow-Glickman (S-G) human gingival epithelial cells were incubated with LPS (1 or 5 µg/ml) ± NDM (high molecular weight non-dialyzable material derived from cranberry juice; 1-500 g/ml) in serum-free medium for ≤6 d. Membrane damage was assessed by lactate dehydrogenase activity released into cell supernatants; viability was assessed by activity of a mitochondrial enzyme. IL-6 and PGE2 were measured by ELISA. Data were analyzed using ANOVA and Scheffe‟s F procedure for post hoc comparisons. Results: Short-term exposure to NDM, or LPS ± NDM caused no significant membrane damage. NDM (≤ 25 g/ml) or LPS ± NDM had no significant effect on viability ≤ 6-d exposure. NDM (≤ 25 g/ml) or LPS ± NDM caused dose-dependent inhibition of production of IL-6 (maximum 70-80%; p≤0.0002) and PGE2 (maximum ~ 60%; p≤0.04). Conclusion: Lack of toxicity of low NDM concentrations to gingival epithelial cells in vitro, 2010 Progress Report Summaries March 25, 2011 Page 14 of 14 and its inhibition of the pro-inflammatory, catabolic mediators PGE2 and IL-6, suggest that cranberry components may regulate oral inflammatory responses and may be useful in prevention or treatment of gingivitis and/or periodontitis. (Supported by UT College of Dentistry Alumni Endowment Fund and by The Cranberry Institute). In addition to the above, we have done very interesting experiments with high molecular weight component of cranberry. We have data demonstrating that NDM arrests the metabolic functions of oral bacteria, for long periods of time, which may be beneficial to the users in preventing oral diseases as a secondary complications to the oral surgical procedures. We have conducted tests with new assay, Live/Dead cell assay and also with flow cytometry. We found that the NDM may not kill bacteria but it is capable of limiting bacterial ability to conduct normal metabolic functions. 2010 Progress Report Summaries March 25, 2011