Progress 10/01/98 to 09/30/04
Outputs
We proposed to develop an immuno-assay approach that will utilize optimum packing of oriented antibodies and combine it with in-line detection devices. For our experiment, a C18 or C16 alky chain with a amine reactive endgroup (carboxyl or NHS) would cross-link with Protein-A. The resulting hydrophobically modified, surfactant like activated protein-A will be allowed to self assemble on the modified hydrophobic substrate. The efficiency of protein-A stacking will be monitored using flurometric measurement of FITC (fluorescein isothiocyanate) anti-Protein-A binding, or by IR. The residual non-antibody covered surface will be subsequently covered with a polyethyleneoxide terminated alkyl chain surfactant through self assembly. The PEO terminus will act to minimize non-specific adsorption of solution proteins and hence lower the background and potential for false signals of the detector. Antibodies (IgG) will be allowed to bind to the self assembled activated protein-A.
Coating of solid surface: Optimization of sialinization was carried out under various times (minutes) of incubation of acid washed glass slides with OTS and drying methods of slides before sialinization. It was observed that drying of glass slides on a hot-plate at 120 C for 15 min before sialinizatin in 1 mM OTS for 15 min works best as observed by a high contact angle of water droplets (102.76 degrees) on sialinized slides. Alkyl chain cross-linking of Protein-A or BSA: A protocol was developed and optimized to cross-link a C16 alkyl chain with a NHS group (C16NHS) with Protein-A and Bovine Serum Albumin (BSA). In brief, excess molar ratio of C16NHS in acetonic solution was added to Protein-A or BSA in phosphate buffer saline (PBS), pH 7.0 to react for 1 hour at room temperature. It was observed that only the reacted protein forms pellet upon centrifugation. This observation was also utilized to remove excess C16NHS from protein solution. Detection of alkyl modified protein by HPLC:
Following alkylation and centrifugal precipitation of BSA and Protein-A, it was dissolved the protein pellets in 0.5% trifluoroacetic acid and injected into C6 HPLC column and run in 0 to 60% acetonitrile gradient. No peaks corresponding to modified proteins were observed, even after prolonged elution. It was concluded that we have experiment with different types of column and solvents to resolve modified protein(s), if any. Self-assembly of alkyl-modified protein on sialinized surface: Various parameters taken into consideration were degree of alkyl modification of protein-A, incubation of modified protein-A in presence or absence of 10% acetone, and washing steps in presence or absence of 1% BSA and/or 0.05% Tween-20. Following assembly, the slides were incubated with Fluorescien Conjugated anti-Protein-A. However, no fluoroscein was detected on the glass slides under the conditions reported above. It was concluded that we should check for efficiency of alkylation of modified
protein and the efficiency self assembly of alkyl chains on sialinized surface before conducting further experiments.
Impacts
The impact of a novel bio-sensor as described in the progress report is yet to be realized. Once the platform technology is developed, applications in solid and liquid foods would be explored.
Publications
- No publications reported this period
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Progress 10/01/02 to 09/30/03
Outputs
Coating of solid surface: Glass slides were acid washed and treated with sialine (octadecyltrichloro silane) in carbon tetra chloride, using a protocol developed at UMaine. The sialinized slides were coated with Poly ethylene glycol (PEG) and checked for passivation efficiency by testing the spread of water droplets and by coating with fluoresceinated bovine serum albumin. The results suggested that sialinization and surface passivation with PEG work in our experiment. Coating various papers: To test surfaces other than glass slides for self-assembly of antibody, the stability of the several papers (Hansol 200, Hansol 80, Copy paper, HP Brochure flyer, Hybond N nylon membrane, and Protran Nitrocellulose membrane, supplied by the Paper Surface Science Program at UMaine) was evaluated against carbon tetrachloride (CCl4) and octadecyl trichlorosilane. The paper samples were subsequently tested the tensile strength following CCl4 treatment. All paper samples were found
suitable for sialinization. Following this evaluation the papers were coated and membranes with sialine for possible use in immunosensor experiments. Subsequently, the papers have been sialinized to be used for immune-assembly experiments. Alkyl chain cross-linking of Protein-A or BSA: A protocol has been developed and optimized to cross-link a C16 alkyl chain with a NHS group (C16NHS) with Protein-A and Bovine Serum Albumin (BSA). Excess molar ratio of C16NHS in acetonic solution was added to Protein-A or BSA in PBS, pH 7.0 to react for 1 hour at room temperature. It was observed that only the reacted protein forms pellet upon centrifugation. This observation was also utilized to remove excess C16NHS from protein solution.
Impacts
If a suitable method can be found to attach antibodies to standard paper surfaces this would have ramifications for novel biosensor platforms using printed paper surfaces.
Publications
- No publications reported this period
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Progress 10/01/01 to 09/30/02
Outputs
Objective (1):Assess technologies to evaluate and ensure post-harvest food quality. (A)Experiments using the FPE model 13500 UV unit showed that the FDA-mandated 5-log10 pathogen reduction in juices can be met with multiple passes through the machine with a total exposure of just over 9 secs.(B) Immunocapture of Escherichia coli (E. coli) in a flow through system, using 3 mm diameter polystyrene beads, was accomplished at a very low capture efficicency (1-2 %). (C) Preliminary experiments show that antibodies will bind to high wet strength bonded paper material in order to begin designing and building paper biosensors for E. coli. (D) Appointment to National Academies of Science Committee the Use of Scientific Criteria and Performance Standards for Safe Food. Objective (2): Determine relationships for models between physical property measurements and nutrition requirements of plant and animal systems.(A) Preliminary experiments relating calcium uptake to bone strength
in simulated microgravity environments are inconclusive at this time. Further experiments are planned in collaboration with the NASA Ames Research Center, Moffet Field, California.
Impacts
1. The effectiveness of a UV machine for cider producers in Maine is being evaluated. If effective, this UV treatment method would provide a low-cost solution to pasteurization for small- and mid-sized apple cider producers. 2. Relationships between food products and the measured physical properties are important as new products are being developed. 3. Research into the development of in-process-line biosensor is very timely and has received significant interest from the food industries and department of defense.
Publications
- Ziegler, C. R., D. W. Donahue, F. A. Drummond, and S. N. Smith. 2002. The Ecological Economics of Insecticide Use Associated with the Maine Potato Industry Based Upon a Producer Survey. Journal of Alternative Agriculture, 17(4):159-166.
- Wentworth, D. S., D. W. Donahue, and R. M. Seymour. 2002. Economic Analysis of Composting Crab Processing Waste. Compost Science and Utilization, 10(1): 47-56.
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Progress 10/01/00 to 09/30/01
Outputs
Objective (1) Experiments using the FPE model 3500 (Oesco, Inc., Conway, MA) of UV pasteurization for apple cider (cider) and like liquid foods were initiated. Sensory evaluation results of the cider (treated with UV) showed that panelists could not discern differences in treated and non-treated cider. Early experiments inoculating cider was inoculated with Escherichia coli (E. coli, ATCC 25922) and treated showed UV treatment was effective to reducing E. coli counts to below detectable levels (using standard plate count methods) for E. coli concentrations of up to 107. The mandated USDA 5-log reduction was not attainable through using just UV treatment, even after up to 4 passes through the machine (exposure time approximately 9.3 s). On average a 3-log reduction was obtained. Further analysis of these findings will be performed. Objective(2) The PI made the Instron 4466 materials testing machine available to two graduate students in food science. The PI assisted
with the design of a plastic tooth for compression tests to simulate the human "bite". The PI assisted in the analysis of their compression test data as it related to calculating strength and texture of food products.
Impacts
1. The usefulness and effectiveness of the UV machine for cider producers in Maine is being evaluated. If effective, this UV treatment method would provide a low-cost solution to pasteurization for small- and mid-sized apple cider producers. 2. Relationships between food products and the measured physical properties are important as new products are being developed.
Publications
- Skonberg, D. I., D. W. Donahue, R. C. Bayer, E. Floreto, and J. G. Riley. 2001. Quality Evaluation of American Lobsters Fed Diets Containing Crab Processing Waste. Journal of Aquatic Food Product Technology, Vol. 10(2):17-29.
- Seymour, R.M., D. W. Donahue, M. Bourdon, J. R. Evans, and D. Wentworth. 2001. Intermittent Aeration for In-vessel Composting of Crab Processing Waste. Compost Science and Utilization, 9(2):98-106.
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Progress 10/01/99 to 09/30/00
Outputs
1. A. Experiments using the FPE model 3500 (Oesco, Inc., Conway, MA) of UV pasteurization for apple cider (cider) and like liquid foods were initiated. Sensory evaluation results of the cider (treated with UV) showed that panelists could not discern differences in treated and non-treated cider. Early experiments inoculating cider was inoculated with E.coli (ATCC 700609) and treated showed UV treatment was effective to reducing E.coli counts to below detectable levels (using standard plate count methods) for E. coli concentrations of up to 103. Further experiments are planned. 2. A. A back propagation neural network (NN) was developed and tested against sensory and compressive force measurements of Maine wild blueberries. The NN was as efficient in prediction of sensory attributes as the compressive force measurements (ca. 72 % efficiency).
Impacts
1. The usefulness of this machine for small cider producers is being evaluated. 2. The neural network will provide a method of classification for blueberries pending future work.
Publications
- Ziegler, C.R., D.W. Donahue, F.A. Drummond, and S.N. Smith. 2000. Agrelation: a computerized decision-making tool for Colorado potato beetle population management and environmental quality concerns. Maine Agricultural and Forest Experiment Station Technical Bulletin No. 176.
- Donahue, D. W. and P. W. Benoit, B. J. Lagasse, and W. R. Buss. 2000. Sensory, Instrumental and Neural Network Evaluation of Maine Wild Blueberries for the Fresh Pack Market. Postharvest Biology and Technology 19: 221-228.
- Benoit, P. W., D. W. Donahue, A. A. Bushway, J. A. Storey, and T. M. Player. 2000. Surfactant Application System to Prevent Anthocyanin Leakage of IQF Blueberries. Journal of Food Quality, 23(3):271-282.
- Long, D. W., F. A. Drummond, E. Groden, and D. W. Donahue. 2000. Modeling Beauveria bassiana Horizontal Transmission. Agricultural and Forest Entomology, 2:19-32.
- Long, D. W., F. A. Drummond, E. Groden, and D. W. Donahue. 1999. Modeling Insect-Pathogen Dynamics. Trends in Entomology, 2:55-62.
- Donahue, D. W., A. A. Bushway, J. M. Smagula, P. W. Benoit, R. A. Hazen. 2000. Assessment of Pre-harvest Treatments on Maine Wild Blueberry Fruit Shelf-life and Processing Quality. Small Fruits Review, 1(1):23-34.
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Progress 10/01/98 to 09/30/99
Outputs
Objective 1. A. In-field application of quality-enhancing sprays studies were completed and analyzed. B. Simulation model development of the Colorado potato beetle and its management is progressing so that a user-friendly simulation model is currently being evaluated by researchers and extension personnel involved with potato production. The model incorporates an environmental economics portion to assess costs associated with different management decisions. C. 1. One flow system has been designed and currently being tested to extract live Escherichia coli cells from apple cider. PI is currently working with a local biosensor research company to further test the effectiveness of the flow extraction. 2. Ultraviolet irradiation is being evaluated as a method to facilitate low temperature pasteurization and ensure Escherichia coli destruction. Objective 2. A. An initial project has been developed to use compression/shear testing to determine strength and stiffness of
horse hooves that have been treated with four hoof-strengthening applications.
Impacts
Objective 1. B. The Colorado potato beetle simulation model will have significant cultural and economic impact on the selection of management alternatives for Maine potato producers. C. In-process-line identification of Escherichia coli will enable fruit juice processors to ensure the safety of their product. UV irradiation will provide a low cost method of pasteurization for small to mid-sized apple cider processors. Objective 2. Horse hooves that are stronger will resist disease invasion.
Publications
- Donahue, D. W., A. A. Bushway, J. M. Smagula, P. W. Benoit, R. A. Hazen. 2000. Assessment of Pre-harvest Treatments on Maine Wild Blueberry Fruit Shelf-life and Processing Quality. Small Fruits Review, 1(1): in press (expected Spring 2000).
- Benoit, P. W., D. W. Donahue, A. A. Bushway, J. A. Storey, and T. M. Player. 1999. Surfactant Application System to Prevent Anthocyanin Leakage of IQF Blueberries. Journal of Food Quality, in press (expected Fall 1999).
- Donahue, D. W., R. C. Bayer, J. G. Riley, A. A. Bushway, P. B. Brown, R. A. Hazen, K. E. Moore, and D. A. deBruyne. 1999. The Effect of Soy-based Diets on Weight Gain, Shell Hardness and Flavor of the American Lobster (Homarus americanus). Journal of Aquatic Food Product Technology, 8(3):69-77.
- Donahue, D. W., D. A. deBruyne, J. D. Fecteau, J. A. Storey, and R.A. Hazen. 1999. Consumer Preference and Mechanical Property Assessment of Maine Wild Blueberries for the Fresh Pack Market. Journal of Food Quality, in press (expected Fall 1999).
- Donahue, D. W., A. A. Bushway, K .E. Moore, and B. J. Lagasse. 1999. Maine Wild Blueberries Field Winnowing Systems. MAFES Technical Bulletin # 174, University of Maine, Orono.
- Donahue, D. W., R. C. Bayer, and M. Loughlin. 1998. Examination of Lead Levels in the American Lobster, Homarus americanus, from Three Sites in Maine. Journal of Shellfish Research,17(4):1247-49.
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Progress 10/01/97 to 09/30/98
Outputs
(1) During 1997, the PI was involved in determining the effects of diets on the American lobster (Homarus americanus). Lobsters in post-molt condition were obtained from a lobster wholesaler. There were four replications on 10 lobsters for each diet trial. Plans for the 1997 study were developed in conjunction with Purdue University, where the soy-based diets were prepared. Four diets, 100% soy, 15% flax seed plus 85% soy, 15% fish meal plus 85% soy, and a cod fish rack control, were used in the study. In these experiments, a relationship was found between weight gain and diet type, a strong relationship between diet and shell strength has been determined. None of the artificial diets performed better than the commonly used diet of codfish racks with respect to weight gain or shell hardness. (2) Simulation model development of the Colorado potato beetle and its management is progressing so that a user-friendly simulation should be available this spring. The model
incorporates and environmental economics portion to assess costs associated with different management decisions. (3) A computer simulation model of an IQF blueberry processing plant was developed to assist management decisions relative to scheduling blueberry processes.
Impacts
(N/A)
Publications
- Donahue, D.W., and T.M. Work. 1998. Sensory and textural evaluation of Maine wild blueberries for the fresh pack market. J. Texture Studies 8(2): 305-312.
- Donahue, D.W., R.C. Bayer, and J.G. Riley. 1998. Effects of diet on weight gain and shell hardness of the new-shelled American lobster, Homarus americanus. J. Applied Aquaculture 8(2): 79-85.
- Donahue, D.W., J.A. Chalmers, and J.A. Storey. 1998. In-vessel compost system evaluation using university food wastes. Compost Science & Utilization 6(2): 75-81.
- Donahue, D.W., R.C. Bayer, T.M. Work, and J.G. Riley. 1997. Effect of diet on weight gain, shell hardness, and flavor of new shell lobsters. J. Applied Aquaculture 7(4): 69-77.
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Progress 10/01/96 to 09/30/97
Outputs
(1) During the 1996-97 year the PI was involved in determining the effects diets on the American lobster (Homarus americanus). Results of the 1996 study were analyzed and plans for the 1997 study were developed. In August 1997, an initial set of experiments was performed using soy-based diets. These results are now being evaluated. (2) A graduate student project determining relationships between aeration rates and material breakdown for in-vessel composting mixtures was completed and the results are being analyzed during Fall 1997. (3) A working computer simulation model of a sawmill has been completed. The model has been verified using data obtained from the management of a local sawmill operation.
Impacts
(N/A)
Publications
- Donahue, Darrell W., R. S. Sowell, and N. M. Bengston. 1996. Economic analysis of U.S. flue-cured tobacco marketing alternatives. Tobacco
- Riley, J. G., D. Donahue, R. Bayer, and G. Ozbay. 1996. Shipping and handling of live lobster. Proc. Conf. on Marketing & Shipping Live Aquatic Products. NRAES Pub. # 107.
- Donahue, D. W., R. C. Bayer, T. M. Work, and J. G. Riley. 1997. The effect of diet on weight gain, shell hardness, and flavor of new-shell American lobsters (Homarus americanus). Journal of Applied
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Progress 10/01/95 to 09/30/96
Outputs
Objective 1: Initial experiments to determine consumer size preference and physical attributes for Maine wild blueberries were performed. Three size classes, based on blueberry diameter, were used; <8 mm, 9-10 mm and 11-12 mm. The large size class blueberries (11-12 mm) were consumer preferred and no significant differences were found int he physical attributes between size classes. Objective 2: Feeding trials on new-shell tidal-pounded lobsters were performed to assess the efficacy of different feeds and their relationship to growth, disease resistance, shell hardness. Weight gain and shell hardness were recorded and analyzed using standard statistical procedures. Results show that, of the diets studied, salted fish racks are the best alternative feed for tidal-pounded new-shell lobsters. A lobster feeding trial, where the lobsters were held in holding tanks, was performed testing the affects of oxygen levels on feed uptake measured by weight gain. No significant
differences in weight gain were found between levels of oxygen applied. Objective 3: The layout of a basic sawmill operation was programmed in ARENA (Systems Modeling Corp., Sewickley, PA) modeling system. Input distribution models for each machine center of the sawmill are being derived.
Impacts
(N/A)
Publications
- Donahue, D.W., R.C. Bayer, T.M. Work, and J.G. Riley. 1996. The effect of diet on weight gain, shell hardness, and flavor of new-shell lobsters. Ann. Meeting Abstracts National Shellfish Assoc., Baltimore.
- Donahue, D.W., R.S. Sowell, and N.M. Bengston. 1996. The U.S. flue-cured tobacco industry, modeling of alternative marketing systems. Agricultural Systems 51(3):1-12.
- Donahue, D.W., R.S. Sowell, and N.M. Bengston. 1996. The U.S. flue-cured tobacco market system, economic analysis of marketing alternatives. Tobacco Science 40(2).
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