Source: NORTH CAROLINA STATE UNIV submitted to
EVALUATION OF THE MILK AND MEAT RESIDUES OF ORGANIC THERAPIES FOR MASTITIS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1005097
Grant No.
2014-51300-22234
Project No.
NC09819
Proposal No.
2014-05326
Multistate No.
(N/A)
Program Code
113.A
Project Start Date
Sep 1, 2014
Project End Date
Feb 28, 2019
Grant Year
2014
Project Director
Anderson, K.
Recipient Organization
NORTH CAROLINA STATE UNIV
(N/A)
RALEIGH,NC 27695
Performing Department
Animal Science
Non Technical Summary
Mastitis, or udder inflammation, is often caused by bacterial infection and is commonly treated using antibiotics. However, organic dairy farmers in the United States are not permitted to use antibiotics, even though organic cows can have mastitis challenges similar to cows on conventional farms. When mastitis occurs, organic farmers use various alternative treatments, many of which are plant-based. Though such treatments are used by organic dairy farmers to help keep their cows healthy, one very important measure has not yet been determined: how long do plant-based products remain in the animal's body? We will evaluate the amount of time that three different organic products for mastitis remain in the milk and meat of both healthy cows and cows with mastitis, and then share this information with organic farmers and organic stakeholders nationwide. We will work with veterinarians, organic dairy farmers, Cooperative Extension personnel, and veterinary students to share the results of our research and increase their knowledge of organic practices. Our research will help organic dairy farmers use plant-based products in a way that prevents residual treatments (such as garlic flavor) from entering the food supply, thereby ensuring the safety and quality of organic milk. Our results may also be applicable to conventional dairy farms as a way to reduce use of antibiotics overall.
Animal Health Component
100%
Research Effort Categories
Basic
20%
Applied
80%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7113440118035%
7113450118035%
3113410110020%
3153410302010%
Goals / Objectives
Educating organic dairy farmers and veterinarians about residue duration and safety of commonly-used organic products for mastitis is the main goal of this project.Objectives:1. Determine if commonly-used organic products for mastitis can trigger antibiotic residue detection tests in milk to assess the potential risk to farmers of using organic treatments.2. Determine the residue duration of commonly-used organic products for mastitis in milk and meat of lactating cows as an essential first step in understanding food safety aspects of such treatments.3. Provide workshops and other educational opportunities for organic dairy farmers and veterinarians to learn about the residue consequences of commonly-used organic products for mastitis.4. Once the milk and meat withdrawal times of these products are established, future studies can be designed to determine the efficacy of such therapies on organic farms across the United States.
Project Methods
Effort: Three organic products for mastitis will be assessed in milk in the laboratory for their ability to trigger an antibiotic residue screening test. These same three products will be administered to healthy lactating cattle at the recommended dose (group 1) and at double the recommended dose (group 2), and milk and blood samples taken from these treated cattle will be analyzed to determine the concentration of the product remaining after a long duration of sampling time points. The products will also be analyzed to determine the molecule present in the most detectable concentration. The molecule of each product that is most detectable will be ordered in radiolabeled form and inserted in cows in groups 1 and 2 following the milk trial. These cows will be euthanized at multiple time points and samples will be collected from various tissues to determine where metabolism of the products likely takes place and the duration of time that residues stay in the tissues post-treatment. Information gained from this project will be shared with the organic dairy farming community and the veterinary and scientific communities through a short course, web seminars, fact sheets, scientific presentations, and peer-reviewed publications.Evaluation: The short course will be evaluated using a voluntary survey following the course with specific questions designed to test the learning objectives of the course as well as space for feedback. Effectiveness of the web seminars and fact sheets will be evaluated by the advisory panel. The development of these outreach materials will lean heavily on the advisory panel for editing assistance and to make sure that the material is appropriate for our target audience. The research science will be evaluated by response to the information at scientific meetings and ability to publish the materials in peer-reviewed journals.

Progress 09/01/14 to 02/28/19

Outputs
Target Audience:The target audience reached by this project includes the national and international scientific community of veterinary, animal and dairy science researchers; the North Carolina State University research community; organic dairy farmers in the Mid-Atlantic United States and across the US; Organic Valley affiliated dairy farmers; veterinary and animal science students at North Carolina State University; and veterinary and animal science biomedical researchers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?As stated before, the project has allowed more than 75 undergraduate and veterinary students to gain insight and experience into the organic dairy industry. Most have gained experience working with dairy cattle for their first time and this has tremendous professional development value. At least 10 undergraduate and veterinary students have gained experience and exposure relative to the conduct of various aspects of research, whether it be seeking funding, writing grants/proposals, designing and conducting hands-on research, writing abstracts and manuscripts, preparing posters, and applying the scientific method. As previously reported, the project has provided professional development for eight undergraduate students who were hired to lead milking shifts and guide the activities of their fellow students. Volunteer involvement with the project has resulted in 65 people from at least 4 countries being trained to work with research on dairy cattle. Two undergraduate students were taught how to write grants and both received the grants they applied for. These grants supported their independent research projects that were directly related to this larger project. An additional undergraduate Honors students has completed a survey of organic producers on mastitis treatment and prevention. Undergraduate and veterinary students were taught to write scientific abstracts and to present at research meetings. At least 2 students from the College of Veterinary Medicine completed projects related to the grant while participating in the Merck Merial Summer Scholars Research Program. Students were also taught how to communicate and work directly with organic dairy stakeholders. Visiting international scientists also participated in various aspects of the project, including Dr. Sara Armorini from the School of Agriculture and Veterinary Medicine, University of Bologna, Bologna, Italy, and a visiting Fulbright Scholar, Dr. Elmer Mehmeti, Head of Sector, Food Safety and Veterinary Institute, Albania. The webinar is a useful resource for training and development of individuals in many phases and aspects of the organic dairy industry. Many additional opportunities were available for providing for education of the public, students and professionals. We had a poster and a booth one year at the Annual College of Veterinary Medicine Open House, thus exposing thousands of people to organic dairy. Project investigators were speakers for various courses in the College of Veterinary Medicine courses and experiential activities (blocks), with the intention of educating students about organic dairy and our project. The issue of treatment for organic dairy and residue issues was presented to students aspart of various teaching laboratories. Our project stimulated a lot of interest, discussion and consideration of organic dairy and issues organicproducers face in dealing with some animal health issues. How have the results been disseminated to communities of interest?Dissemination of the results has been via refereed publications, presentations at national and international scientific meetings, at producer and extension meetings, and to organic producers. The US Food Animal Residue Avoidance Databank has used the information to inform their personnel in terms of providing science-based recommendations to producers and veterinarians. The cited published manuscripts and presentations are the primary manner in which the results have been disseminated to the research community. Results of sections of the larger objectives have been shared with the scientific community at national and international veterinary research conferences, at Annual Dairy Science meetings, at the North Carolina State University Undergraduate Research Symposium, at the Merck Merial Summer Scholars Symposium, at the Annual North Carolina State University College of Veterinary Medicine Research Forum and at the Vermont Organic Dairy Conference, as well as informally to organic dairy farmers in North Carolina, Virginia, and West Virginia while visiting their farms to find cows to purchase to complete the project. An advisory committee comprised of organic industry stakeholders, researchers, and farmers formed in the first year and then met twice via webinar to provide input and to be updated. The comprehensive concluding webinar on the overall outcome of the project will provide continuing education for our peers, allied professionals, all levels of the organic dairy industry and others. It will be available via Moodle Software from North Carolina State University. Experiential Learning: During the entire project, multiple avenues for experiential learning were available for students at various levels. Some examples follow: Throughout the whole study (2014-2017), Dr. Mullen took students to organic dairy farms in WV, VA and NC to inspect cattle for potential inclusion in the project. This included 15 different students (mainly undergraduates). Students were then coached on how to explain the research being conducted to the farmers. Sept 27, 2016 - Organic Valley Pasture Walk and field day -Little Brown Cow Dairy, Monroe Co. WV - attended by Drs. Mullen and Washburn. A total of more than 60 laypeople were in attendance - local farmers and extension personnel from 3 states. See additional material on Dissemination under the section on Dissemination of Results. The comprehensive concluding webinar on the overall outcome of the project will provide continuing education for our peers, allied professionals, all levels of the organic dairy industry and others. It will be available via Moodle Software from North Carolina State University. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? What was accomplished under these goals? Impact: Producers in the growing US organic dairy industry encounter mastitis in their herds and need access to treatments for mastitis that are safe and effective with minimal risk of milk and meat residues. Mastitis occurs as commonly on organic as conventional dairies but organic producers cannot use synthetic antibiotics. Organic producers desire to address the health and animal welfare concerns of their animals. There is a lack of approved and safe products for management of udder health on organic dairies. Products available include plant-based or herbal products (phytoceuticals) for maintaining udder health. Unfortunately, little research has considered how best to use these products, especially in terms of how long their ingredients remain in milk, meat, and tissues of the treated animal. Our objective was to advance knowledge relative to appropriate and safe use of these products, especially in terms of residues in milk and meat. The first step in addressing this was to develop accurate, reliable and reproducible detection methods that measure indicator molecules of the organic products. Extensive effort was put into developing methods that accurately measure the indicator molecules. These methods were developed and published to accurately detect concentrations of indicator molecules of 3 commercially available phytoceuticals. In a series of experiments, organic dairy cows exposed to different doses of the 3 phytoceuticals were repeatedly sampled and concentrations of indicator molecules were measured. Analysis produced estimates that will help dairy producers and veterinarians use these products in a way to avoid residues in marketed products from the treated cows. Objective 1: Determine if phytoceuticals cause false positive tests or interfere with residue detection tests: We have evaluated commonlyused milk antibiotic residue tests: the Delvotest SP and Charm SLBL tests. Research has been completed and published regarding cross-reaction of the 3 organic compounds tested with the two different tests and their ability to cause positive tests. We concluded that phytoceutical products in bovine milk at physiologic levels would not cause positive test results for the Delvotest SP and would not interfere with the Charm SLBL and other Charm tests in detection of various antibiotics. See Products-Publications. Objective 2: Measure and assess presence of phytoceutical indicator molecules in milk, blood and tissues following administration of phytoceuticals: Three major accomplishments have been achieved: Analytical methods: Development, validation and performance of analytical methods to detect indicator compounds for these products. Animal studies: A total of 110 lactating dairy cattle were maintained and milked under organic conditions (except for access to pasture while in the research facility). Cows were administered the various treatments at different doses using the routes specified by the product labels. Samples of milk, blood (plasma), and other matrices were collected over time following administration of the test doses. 3. Analysis of drug concentration data over time: Statistical and pharmacokinetic methods were used to study the drug concentrationsdetermined (see #1 and 2 above) over time and to produce the best description of drug disappearance over time. Samples were collected of plasma, body tissues, and milk for residue studies. Fifty healthy cattle were dosed with three different products. Sixty cattle were used to evaluate the plasma and milk residues from cows with natural or induced mastitis infections. Thirty cattle were purchased from organic farms with subclinical mastitis (high SCC, no visible changes in milk) and 30 additional cattle free of infection were challenge-inoculated with Streptococcus uberis to cause mastitis. When challenged cows developed clinical mastitis they were administered phytoceuticals. Samples of plasma and milk were collected for chemical analysis and additional microbiological and somatic cell count analyses for milk. Chemical analysis of the tissue, plasma and milk samples has been completed for thymol, carvacrol, and diallyl disulfide (the "indicator molecules" mentioned previously). Pharmacokinetic analysis of the healthy tissues and plasma has been completed for all 3 compounds and the results were published in 2017--see Products-Publications. Because none of the products tested nor their active ingredients are approved by FDA for use in dairy cattle, active compounds should not be detected in tissues. Based on our pharmacokinetic data and FDA requirements for no detection, we estimate that use of FDA methods would produce the following estimated meat withholding times (time after last dose for ingredients to decrease to levels below detection) for the products as formulated when we tested them: 0 days for Dr. Paul's CEG Tincture, 10 days for Uddersol™, and 13 days for Phyto-Mast®. Because the herbal preparations commonly used are generally regarded as safe (GRAS), some individuals may suggest that shorter times may be considered. Our study did not attempt to address this controversy, but rather to provide sound scientific information that could be used to inform any consideration of residue potential. Pharmacokinetic analysis in milk is complete and published for thymol and carvacrol and for comparing diseased and healthy cattle.This has also been published--see Products-Publications. Objective 3: Dissemination of results and providing workshops and educational outreach: See separate section following. Objective 4: Design additional and future studies on efficacy of the products The research completed for this project was an initial examination of the residue potential for just three phytoceuticals. Several additional projects, listed below, were completed during the project to enhance our understanding of phytoceuticals. However, there are many more phytoceutical products on the market for organic dairy cattle that would benefit from similar pharmacokinetic studies. Milk cellular changes over time have been analyzed in cows treated with phytoceuticals and normal cows have been compared to cows with mastitis. Resulted in student abstract at a research conference and at a national/international meeting: see Products section. Samples from the clinically induced Streptococcus uberis mastitis samples have allowed an evaluation of the efficacy of the 3 phytoceutical products to eliminate clinical Strep. uberis mastitis. This has resulted in the publication of a manuscript in the Journal of Dairy Science--see Products-Publications. We used the data from the healthy cow study to investigate the ability of intramammary thymol and carvacrol to cross the mammary-blood barrier and secured supplementary funding from Organic Valley Cooperative. This resulted in extensive research training for 2 students and data that were published in an Organic Valley report and pamphlet that can be disseminated to organic dairy producers. The cattle from the study were used to develop techniques involving novel interstitial probe placement in the udder and determination of tissue levels of flunixin meglumine (an anti-inflammatory drug that can be used by organic dairy farmers). Those data have been analyzed and were presented in October, 2017-- see Products. An undergraduate Honors student surveyed organic producers to provide quantitative data on mastitis treatment and prevention techniques used by US organic producers. An on-line survey was conducted using Organic Valley producers. We did learn during the conduct of the project that many producers don't have on-line access, especially in the Midwest, and that needs to be considered in planning programs to reach such producers. Survey responses were analyzed and a final report produced. The results are presented in the webinar.

Publications

  • Type: Other Status: Published Year Published: 2017 Citation: Worsley, K., K.A.E. Mullen, S.E. Mason, K.L. Anderson, S.P. Washburn and R.E. Baynes. Crossover data in milk of cows dosed with Phytomast. Organic Valley Project Report. April, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Mzyk, D., K. Mullen, G. Hobgood, R. Baynes, D. Foster. Continuous samples of health and mastitis quarters of lactating cattle by ultrafiltration for pharmacokinetic modeling. European Buiatrics Forum, Bilbao, Spain.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: K. A. E. Mullen, R. L. Lyman, S.P. Washburn, R.E. Baynes, and K. L. Anderson. 2018. Short Communication: Effect of three phytoceutical products on elimination of bacteria in experimentally-induced Streptococcus uberis clinical mastitis. Journal of Dairy Science. 2018 Nov;101(11):10409-10413. doi: 10.3168/jds.2017-14279. Epub 2018 Aug.


Progress 09/01/16 to 08/31/17

Outputs
Target Audience:The target audience reached by this project to date includes the national and international scientific community of veterinary, animal and dairy science researchers; the North Carolina State University research community; organic dairy farmers in the Mid-Atlantic United States and across the US;Organic Valley dairy affiliated dairy farmers;veterinary and animal science students at North Carolina State University; and veterinary and animal science biomedical researchers. Changes/Problems:The major change encountered this year has been personnel changes. Dr. Keena Mullen has moved into another position effective in the early part of this year. Dr. Steve Washburn has retired effective June 30. However, we are fortunate that both remain available and committed to a succesful end to the Project. What opportunities for training and professional development has the project provided?As stated before, the project has allowed more than 50 undergraduate and veterinary students to gain insight and experience intothe oerganic dairy industry. Most have gained experience working with dairy cattle for their first time and this has tremendous professional development value. At least 10 undergraduate and veterinary students have gained experience and exposure relative to the conduct of various aspects of research, whether it be seeking funding, writing grants/proposal, designing and conducting hands-on research, writing abstracts and manuscripts, preparing posters, and applying the scientific method. As previously reported, the project has provided professional development for eight undergraduate students who have been hired to lead milking shifts and guide the activities of their fellow students. Volunteer involvement with the project has resulted in 65 people being trained to work with research dairy cattle. Two undergraduate students were taught how to write grants and both received the grants they applied for. These grants will support their independent research projects that are directly related to this larger project. An additional undergraduate Honors students has completed a survey of organic producers on mastitis treatment and prevention. Undergraduate and veterinary students were taught to write scientific abstracts and to present at research meetings. Students were also taught how to communicate and work directly with organic dairy stakeholders. How have the results been disseminated to communities of interest?The cited published manuscripts and presentations are the major manner in which the results have been disseminated to the research community. Results of sections of the larger objectives have been shared with the scientific community at national and international veterinary research conferences, at the recent Annual Dairy Science meeting, at the North Carolina State University Undergraduate Research Symposium, and at the Vermont Organic Dairy Conference, as well as informally to organic dairy farmers in North Carolina, Virginia, and West Virginia while visiting their farms to find cows to purchase. We are currently planning a webinar on the overall outcome of the project to present as Continuing Education for our peers, allied professionals and others. What do you plan to do during the next reporting period to accomplish the goals?All animal experiments have been completed during this current year. Analyses of all samples will be completed by the end of 2017. We have an extensive amount of data to be analyzed and presented in publications. A major task for the upcoming year will be dissemination of the results. Thus, the plan for the coming year includes: 1. Enter, double-check and analyze all data collected. 2. Perform appropriate statistical and pharmacokinetic analyses on all data. 3. Study the results of analysis and formulate/synthesize the major findings. 4.Present/publish/disseminate important results from the study. 5. Develop/plan additional research and investigation.

Impacts
What was accomplished under these goals? Impact: Producers in the growing US organic dairy industry need access to treatments for mastitis that are safe, effective and that leave no residuals in the cowand her milk. Mastitis occurs as commonly on organic as conventional dairies butorganic producers cannot use synthetic antibiotics. Organic producers desire to address the health and animal welfare concerns of their animals. There is a lackof approved and safe products for management of udder health. Products available include herbal products (phytoceuticals) for maintaining udder health. Unfortunately, little research has considered how best to use these products, especially in terms of how long indicator molecules stay in the animal and its products. The objective is to advance knowledge relative to appropriate and safe use of these products, especially in terms of residuals in milk and meat. Methods were developed and publishedto accurately detect concentrations of indicator molecules of 3 commonly used phytoecuticals.In a series of experiments, organic dairy cows exposed to different doses of the 3 phytoecuticals were repeatedly sampled and concentrations of indicator molecules were detemined. Analysis produced estimates that would help dairy producers and veterinarians use these products without producing residues. Dissemination of the resultshas been via refereed publications, presentations at national and international scientific meetings, at producer and extension meetings, and to organic producers. The US Food Animal Residue Avoidance Databank has used the information to inform their personnel in terms of providing science-based recommendations to producers and veterinarians. Objective 1: Determine if phytoceuticals cause false positive tests or interfere with residue detection tests: We have evaluated two commonly used milk antibiotic residue tests, the Delvotest SP and Charm SLBL tests. Research has been completed regarding cross-reaction of the 3 organic compounds tested with the two different tests and their ability to cause positive tests. We concluded that phytoceutical products in bovine milk at physiologic levels would not cause positive test results for the Delvotest SP and would not interfere with the Charm SLBL and other Charm tests in detection of various antibiotics. See Products-Publications. Obj 2: Measure and assess presence of phytoceutical indicator molecules following treatment with phytoceuticals: Three major accomplishments have been achieved: 1. Analytical methods: Development, validation and performance of analytical methods to detect indicator compounds for these products. 2. Animal studies: A total of 110 lactating dairy cattle were maintained and milked under organic conditions. Cows were administered the various treatments by different routes anddoses. Samples of milk, blood, and other matrices were collected over time following administration of the test doses. 3. Analysis of drug concentration data over time: Statistical and pharmacokinetic methods were used to study the drug concentrations determined (see #1 and 2 above) over time and to produce the best description of drug disappearance over time. Sample collection from plasma, body tissues, and milk for residue studies has been completed. Fiftyhealthy cattle weredosed with three different products. Sixty cattle were used to evaluate the plasma and milk residues from cows with natural or induced mastitis infections. Thirty cattle were purchased from organic farms with subclinical mastitis (high SCC, no visible changes in milk) and 30 additional cattle free of infection were challenge-inoculated with Strep. uberis to cause mastitis. Mastitis cattle were treated with phytoceuticals.Samples of plasma and milk were collected for chemical analysis and additional microbiological and somatic cell count analysis for milk. Chemical analysis of the tissue, plasma and milk samples hasbeen completed for thymol,carvacrol, and diallyl disulfide. Pharmacokinetic analysis of the healthy tissues and plasma has been completed for all 3 compounds and the results were published in 2017--see Products-Publications. Because none of the products tested nor their active ingredients are approved by FDA for use in dairy cattle, active compounds should not be detected in tissues. Based on our pharmacokinetic data and FDA requirements for no detection, we estimate that use of FDA methods would produce the following estimated meat withholding times: 0 days for Dr. Paul's CEG Tincture, 10 days for Uddersol™, and 13 days for Phyto-Mast®. Because the herbal preparations commonly used are generally regarded as safe (GRAS), some individuals may suggest that shorter times may be considered. Pharmacokinetic analysis in milk is complete for thymol and carvacrol and forcomparing diseased and healthy cattle. The analysis of the pharmacokinetics of milk in both diseased and healthy cattle is in preparation for publication. Additional statistical analysis is being performed, including a comparison of healthy and diseased cattle, with an emphasis on the pharmacokinetics ofclinical vs. subclinical mastitis. Objective 3: Dissemination of results and providing workshops and educational outreach: An advisory committee comprised of organic industry stakeholders, researchers, and farmers formed in the first yearmet twice via webinar to provide input and be updated. Experiential Learning: Throughout the whole study (2014-2017), Dr. Mullen took students to organic dairy farms in WV, VA and NC to inspect cattle. This included 15 different students (mainly undergraduates). Students were then coached on how to explain the research being conducted to the farmers. Sept 27, 2016 - Organic Valley Pasture Walk and field day -Little Brown Cow Dairy, Monroe Co. WV - attended by Drs. Mullen and Washburn. A total of more than 60 laypeople were in attendance - local farmers and extension personnel from 3 states. See additional material on Dissemination under the section on Dissemination of Results. Objective 4: Design additional and future studies on efficacy of the products 4. Initial studies should provide residue data for consideration in of times to zero concentrations of active ingredients inmilk and meat and to inform other relevant studies. Relevant "spin-off" studies are noted below. A. Milk cellular changes over time have been analyzed in cows treated with phytoceuticals and normal cows have been compared to cows with mastitis. Resulted in student abstract at a research conference and at a national/international meeting: see Products section. B. Samples from the clinically induced Strep. uberis mastitis samples has allowed an evaluation of the efficacy of the 3 phytoceutical treatments to eliminate Strep. uberis mastitis. This has resulted in the preparation of a manuscript to be submitted to the Journal of Dairy Science. C. We used the data from the healthy cow study to investigate the ability of intramammary thymol and carvacrol to cross the mammary-blood barrier and secured supplementary funding from Organic Valley Cooperative. This resulted in training for 2 students and data that were published in an Organic Valley report and pamphlet that can be disseminated to organic dairy producers. D. The cattle from the study were used to developtechniques involving novel interstitial probe placement in the udder and determination of tissue levels of flunixin meglumine. Those data have been analyzed and werepresented in October, 2017--see Products. E. An undergraduate Honors student is surveying organic producers to provide quantitative data on mastitis treatment and prevention techniques used by US organic producers. An on-line survey was conducted using Organic Valley producers. However, many producers don't have on-line access, especially in the Midwest, and that needs to be considered in planning programs to reach such producers. Survey responses are being analyzed and will be summarized soon.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Keena AE Mullen, Erin Beasley, Julio Q Rizzo, Steven P Washburn, Ronald E Baynes, Sharon E Mason and Kevin L Anderson. (2017). Potential of phytoceuticals to affect antibiotic residue detection tests in cow milk in a randomized trial. Veterinary Record Open. 4(1): 1-7. doi:10.1136/ vetreco-2016-000214.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Armorini S, J Yeatts, KAE Mullen, SE Mason, E Mehmeti, KL Anderson, SP Washburn, and RE Baynes. 2016. Development and validation of HS-SPME-GC/MS/MS method for the quantification of thymol and carvacrol in bovine matrices and application to pharmacokinetic studies. Journal of Agricultural and Food Chemistry. 64 (41), pp 78567865.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Mason SE., Mullen, K.A.E., Anderson, K.L., Washburn, S. P., Yeatts, J.E.Baynes, R.E. 2017. Pharmacokinetic analysis of thymol, carvacrol and diallyl disulfide after intramammary and topical applications in healthy organic dairy cattle. Journal of Food Additives and contaminants: Part A. 34(5): 740-749.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Mullen, K.A.E. 2017. Considerations for managing mastitis and milk quality on organic dairy farms. ADSA Southern Section Symposium, Abstract #153, Annual Meeting of American Dairy Science Association, June 25-28, 2017, Pittsburgh, PA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Womble, C. M., K. A. E. Mullen, S. P. Washburn, and K. L. Anderson. 2017. Characterization of milk cellular response to intramammary phytoceutical administration in cows with clinical mastitis. J. Dairy Sci.:100, E-Suppl 2:, p 186. (Abstr. 168). https://www.adsa.org/2017/abstracts.asp


Progress 09/01/15 to 08/31/16

Outputs
Target Audience:The target audience reached by this project to date includes the North Carolina State University research community, 11 organic dairy farmers in the Mid-Atlantic United States, students at NCSU, and veterinary biomedical researchers. In addition, twenty-five special-needs high school students were taught about dairy cattle during this reporting year. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project has provided professional development for eight undergraduate students who have been hired to lead milking shifts and guide the activities of their fellow students. Volunteer involvement with the project has resulted in 65 people being trained to work with research dairy cattle, including individuals from 5 different countries and 53 undergraduate and graduate students. Two undergraduate students were taught how to write grants and both received the grants they applied for. These grants will support their independent research projects that are directly related to this larger project. How have the results been disseminated to communities of interest?Results of sections of the larger objectives have been shared with the scientific community at an international veterinary research conference, at the North Carolina State University Undergraduate Research Symposium, and at the Vermont Organic Dairy Conference as well as informally to organic dairy farmers in North Carolina, Virginia, and West Virginia while visiting their farms to find cows to purchase. What do you plan to do during the next reporting period to accomplish the goals?We will continue to collect data from the remaining 20 cows, then analyze those data and submit the results to be published in peer-reviewed scientific journals. We will organize workshops and seminars to disseminate the information from the grant to organic dairy producers and veterinarians. We will also use the data obtained in this project as leverage to design future studies to evaluate the efficacy of these products on-farm.

Impacts
What was accomplished under these goals? Data from Objective 1 have been analyzed and are in a draft scientific manuscript ready for submission. For Objective 2, 100 of the 120 cattle have been evaluated for the project and residue duration in healthy cows has been established. These data are going into scientific manuscripts in preparation. For Objective 3, we plan to host workshops and training opportunities in the coming year. During the current reporting period, over 100 organic dairy farmers in Vermont were in the audience of a presentation about herbal products for mastitis usage given by Keena Mullen, and that stimulated good discussion to develop additional educational resources for farmers. Two studies have examined further aspects of the current project and we will be working on developing future studies - and grant proposals - in the coming months. A poster update was presented at the OREI PI meeting in Washington, DC in October, 2015.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Baynes, RE., Mason, SE., Mullen, K., Anderson, K., and Washburn, K. (2016). Chemical residues in dairy cattle following use of organic therapies for mastitis. Dublin, Ireland. July 2-9th., 2016. World Buiatrics Congress, Dublin, Ireland. July 2-9th., 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Baynes, RE., Mason, SE., Mullen, K., Anderson, K., and Washburn, K. (2016). Residues following organic therapies in livestock. American College of Veterinary Internal Medicine (ACVIM) Forum, Denver, CO. June 8th  11th, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Fitton, AA, Nixon, E, Brooks, JD., Yeatts, J, Mullen, KAE., Washburn, S, Baynes, RE. (2016). Preliminary assessment of thymol and carvacrol partitioning in bovine cream and skim milk fractions. 2016 Annual Summer Undergraduate Research Symposium, North Carolina State University, Raleigh, NC. August 2nd, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Pujianto, C, Yeatts, J, Mullen, K., Washburn, S., Baynes, RE. (2016). Chemical Analysis of Plant Based Mastitis Treatment for Organic Dairy Cows. 2016 Annual Summer Undergraduate Research Symposium, North Carolina State University, Raleigh, NC. August 2nd, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Womble, C., Anderson, KL, Mullen, KAE, Baynes, RE. (2016). Evaluation of milk cellular response to phytoceuticals in cows with mastitis. 2016 Annual Summer Undergraduate Research Symposium, North Carolina State University, Raleigh, NC. August 2nd, 2016.


Progress 09/01/14 to 08/31/15

Outputs
Target Audience:The target audience includes organic dairy producers and the organic dairy industry, including veterinarians and other professionals working directly or indirectly with organic dairy production systems. To date, the project has reached and/or provided preliminary information to members of the North Carolina State University (NCSU) research community, several organic dairy farmers in the Mid-Atlantic United States, organic dairy farmers in other regions of the country, students at NCSU, veterinary biomedical researchers, and an advisory group consisting of organic dairy producers, organic researchers, and others with interest in organic dairying. Two aspects of the project have been presented at the NCSU Undergraduate Research Symposium and at the Merial Scholars Program National Symposium in Davis, CA. Also, general information about the project was presented at an open house at NCSU College of Veterinary Medicine that included hundreds of visitors including potential students and their families. Changes/Problems:Our rationale for using the antibiotic residue screening tests Delvotest SP-NT, Charm CowSide II, and Penzyme was that these are tests that the farmer can use in the milking parlor to instantly test the milk. However, with further discussion, we decided to use two tests used by regulatory agencies because of their increased sensitivity and because they are used at the processing plant to ultimately determine if the milk is acceptable for human consumption. The tests we are now using are the Charm SLBL test and Delvotest P. We will still be able to detect a variety of antibiotic drugs from different antibiotic classes, and the data from this project will be of interest to regulators as well as farmers. We changed our approach to measuring the meat residues of organic products from using radiolabeled molecules to using the whole product, as we are doing in the milk residue trial. There were several reasons for this change in approach; the most important reason is that handling the waste from the animals dosed with radiolabeled molecules was not feasible. With this change in approach, we are able to measure our target molecules (thymol, carvacrol and diallyl disulfide) in each of the herbal products and we now have the ability to measure other active ingredients in the products that would not be radiolabeled. This is important for future research to ensure that all herbal compounds are cleared from each cow's system before milk or meat from that animal is sold for human consumption. We also plan to use whole herbal products rather than radiolabeled molecules when we study cows with mastitis infections next year. What opportunities for training and professional development has the project provided?The project has provided professional development for six undergraduate students who have been hired to lead milking shifts and guide the activities of their fellow students. Volunteer involvement with the project has resulted in 48 people being trained to work with research dairy cattle, including individuals from 4 different countries and 37 undergraduate and graduate students. This project has provided material for 2nd year NC State veterinary medicine student Vanessa Wolf to participate in the NCSU CVM Merial Veterinary Scholars Program under the direction of Dr. Anderson. The project provided a training opportunity for a visiting international Marco Polo Scholar, Dr. Sara Armorini, from the School of Agriculture and Veterinary Medicine, University of Bologna, Bologna, Italy and for visiting international Fulbright Scholar, Dr. Elmira Mehmeti, Head of Sector, Food Safety and Veterinary Institute, Albania. How have the results been disseminated to communities of interest?Preliminary results of sections of the larger objectives have been shared with the scientific community at a national veterinary research conference, at the North Carolina State University Undergraduate Research Symposium, and the March 28th, 2015 NCSU CVM Annual Open House at NCSU-CVM, Raleigh, NC. Preliminary project results have also been presented informally to organic dairy farmers from several states during a farm tour and during the cattle selection process for various replications of the experiment. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue collecting and analyzing data to address Objectives 1 and 2. A follow-up study is planned related to Objective 1. For Objective 2, the final 20% of data related to milk and meat residues will be collected in late 2015 and another series of replicates will be conducted in 2016 to examine the kinetics of milk residues in cows with active udder infections.

Impacts
What was accomplished under these goals? Preliminary data collection is complete for Objective 1, to determine if organic products for mastitis can trigger antibiotic residue detection tests. A follow-up study is being planned. For Objective 2, 80% of the research has been completed to determine the residue duration of organic products for mastitis in milk and meat of healthy cattle receiving the various treatments. Another set of replicates is planned for 2016 to examine if the milk residue kinetics are similar for cows that have udder infections.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: Evaluating milk cellular response to intramammary and topical herbal products. V. Wolf, S. Washburn, R. Baynes, K. Mullen, M. Correa, and K. Anderson. Presented by V. Wolf at the 2015 Merial-NIH National Veterinary Scholars Symposium July 30-August 2, 2015 in Davis, CA.
  • Type: Other Status: Other Year Published: 2015 Citation: "Do phytoceutical products used for maintaining udder health in dairy cows trigger positive reactions from milk antibiotic residue tests?" Erin Beasley. North Carolina State University Honors Capstone Project. 2015.