GOAT DAIRY FOODS USING FUCOXANTHIN AS AN ANTI-OBESITY BIOFUNCTIONAL INGREDIENT

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: OTHER GRANTS
• Reporting Frequency: Annual
• Accession No.: 1018116
• Grant No.: 2019-38821-29037
• Cumulative Award Amt.: $299,913.00
• Proposal No.: 2018-04977
• Project Start Date: Jan 1, 2019
• Project End Date: Dec 31, 2021
• Grant Year: 2019
• Program Code: [EQ]- Research Project

Recipient Organization
PRAIRIE VIEW A&M UNIVERSITY
P.O. Box 519, MS 2001
PRAIRIE VIEW,TX 77446

Performing Department
CARC

Non Technical Summary
Consumers have an increased interest in nutrition as foods have been linked to health and wellness, primarily in the control of obesity, which constitutes a substantial risk for hypertension, type-2 diabetes,cardiovascular diseases and certain cancers. One third of the adult population in US is obese and 17% of children suffer from obesity. Since obesity continues to be an important problem, developing effective preventive measures to reduce obesity and ease the medical and economic burden of these diseases, which amounts to billions of dollars annually on the population, is essential. Fucoxanthin has been reported to exhibit anti-obesity, anti-diabetic, and anti-inflammatory activities. This research proposal will assess the effect of supplementation of fucoxanthin, an anti-obesity biofunctional ingredient, on the quality and acceptability of goat dairy foods. Four goat dairy products (milk, yogurt, cheese, and ice cream) will be prepared containing fucoxanthin as a supplement at 2.4 mg per serving size, which has been shown to be the minimum concentration for anti-obesity effect in human studies. The physicochemical and sensory properties, and acceptability of goat dairy products with fucoxanthin will be studied immediately after production and during storage. Successful supplementation of fucoxanthin into stable and acceptable quality goat dairy products with health benefits will support the USDA-NIFA priority area of health and nutrition affecting the well-being of the population. Thus, this proposal may play a significant role in enhancing advanced research in healthy goat dairy products and can open up avenues of market opportunities for limited resource producers and the food industry. .

Animal Health Component

30%

Research Effort Categories

Basic

20%

Applied

30%

Developmental

50%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
501	3450	1010	40%
502	3430	1010	30%
502	3460	1010	30%

Knowledge Area
501 - New and Improved Food Processing Technologies; 502 - New and Improved Food Products;

Subject Of Investigation
3430 - Cheese; 3450 - Milk; 3460 - Ice cream;

Field Of Science
1010 - Nutrition and metabolism;

Keywords

goat milk

anti-obesity

biofunctional

fucoxanthin

products

Goals / Objectives
Consumers have an increased interest in nutrition as foods have been linked to health and wellness, primarily in the control of obesity, which constitutes a substantial risk for hypertension, type-2 diabetes, cardiovascular diseases and certain cancers. One third of the adult population in US is obese and 17% of children suffer from obesity. Since obesity continues to be an important problem, developing effective preventive measures to reduce obesity and ease the medical and economic burden of these diseases on the population is essential. Fucoxanthin has been reported to exhibit anti-obesity, anti-diabetic, and anti-inflammatory activities. This research proposal will assess the effect of supplementation of fucoxanthin, an anti-obesity biofunctional ingredient, on the quality and acceptability of goat dairy foods. Four goat dairy products (milk, yogurt, cheese, and ice cream) will be prepared containing fucoxanthin as a supplement at 2.4 mg per serving size, which has been shown to be the minimum concentration for anti-obesity effect in human studies. The physicochemical and sensory properties, and acceptability of goat dairy products with fucoxanthin will be studied immediately after production and during storage. Successful supplementation of fucoxanthin into stable and acceptable quality goat dairy products with health benefits will support the USDA-NIFA priority area of health and nutrition affecting the well-being of the population. Thus, this proposal may play a significant role in enhancing advanced research in healthy goat dairy products and can open up avenues of market opportunities for limited resource producers and the food industry.The overall objective of this project is to develop and evaluate the sensory, physicochemical characteristics, and acceptability of goat dairy products containing fucoxanthin as an anti-obesity biofunctional ingredient. Successful implementation of this project will support the NIFA-CBG Program priority area of human health and obesity as it relates to nutrition. This will be accomplished in part by the development and evaluation of nutritious and healthy value-added goat dairy products from Alpine goats raised on the PVAMU Farm. The specific objectives of this study are to: (a) develop and evaluate sensory and physicochemical characteristics of goat milk containing fucoxanthin after production and during storage at 4°C for 3 weeks; (b) develop and evaluate sensory and physicochemical characteristics of goat milk yogurt containing fucoxanthin after production and during storage at 4°C for 3 months; (c) develop and evaluate sensory and physicochemical characteristics of goat cheese containing fucoxanthin after production and during storage at 4°C for 3 months; (d) develop and evaluate sensory and physicochemical characteristics of goat milk ice cream containing fucoxanthin after production and during storage at -26°C for 3 months.

Project Methods
We propose to develop and evaluate the sensory, physicochemical properties, and acceptability of four goat dairy products (pasteurized milk, yogurt, cheese, and ice cream). Raw whole goat milk will be obtained from the International Goat Research Center (IGRC) at PVAMU. A normal pasteurization technique that is used in the industry will be applied to raw goat milk (65°C x 30 min). Prior to the pasteurization and homogenization steps, fucoxanthin will be added to milk such that the final concentration in the product is equivalent to 2.4 mg in 250 ml of milk, which is one serving size (Mok et al., 2016). This concentration is also the minimum level of fucoxanthin demonstrated to have an anti-obesity effect in humans (Abidov et al., 2010) and will be added to all of goat dairy products. After successful completion of fucoxanthin supplementation in goat milk, the team will devote efforts to determine the stability and degradation rate of fucoxanthin-casein complex during the storage of milk. Goat milk without fucoxanthin (control) and goat milk supplemented with fucoxanthin (treatment) will be evaluated for sensory attributes, acceptability, and storage stability according to the outlined protocol (Figure 1, Appendix) of the proposal.Goat milk yogurt will be manufactured from whole milk with the addition of powdered goat milk to constitute 14.4% total solids (TS) to simulate commercial regular yogurt. Freeze-dried lactic acid culture containing Lactobacillus bulgaricus, Lactobacillus thermophilus, and Lactobacillus acidophilus will be added to the final mix according to the method of Attaie et al. (1987). The mix will be fermented in a culture tank at 40oC and interrupted by cooling when the pH value of yogurt reaches 4.2 to 4.3. The fucoxanthin content will be added such that the final concentration in the product is equivalent to 2.4 mg fucoxanthin/170 g of yogurt (1 serving size = 1 container, 170 g). Yogurt without fucoxanthin (control) and with added fucoxanthin (treatment) will be evaluated for sensory attributes, acceptability, and storage stability.Soft goat cheese (Chevre) will be manufactured according to the method described in the book "Fabrication of Farmstead Goat Cheese" (Le Jaouen, 1990). Likewise, goat milk cheese will be supplemented with fucoxanthin to form fucoxanthin-casein complex in the cheese matrix. The adjunct Lactobacillus casei variant A (Figure 2, Appendix) will be used to favor not only the development of the organoleptic characteristics of soft goat milk cheese, but also the formation of the amino acids tryptophan (Trp) and tyrosine (Tyr), which are involved in the binding of fucoxanthin to casein. The quantity of fucoxanthin will be added such that the final concentration in the product is equivalent to 2.4 mg of fucoxanthin/28 g of cheese (1 serving size = 1 slice, 28g). Soft goat cheese without fucoxanthin (control) and with added fucoxanthin (treatment) will be evaluated for sensory attributes, acceptability, and storage stability.Goat milk ice cream will be made according to the method that was developed at the PVAMU creamery (Attaie; unpublished data). Goat milk ice cream made at the PVAMU creamery was popular on campus and had the following composition: 12% fat, 11% milk solids not fat (MSNF), 15% sugar, 0.3% stabilizer, and 38.3% total solids. According to the marketability survey conducted at the University campus, the goat milk chocolate and vanilla ice creams were scored high and liked very much. Fucoxanthin will be added to ice cream mix such that the final concentration in the product is equivalent to 2.4 mg fucoxanthin per 4 fl. oz of ice cream (1 serving size = ½ cup, 4 fl. oz). Vanilla ice cream made from goat milk without fucoxanthin (control) and with added fucoxanthin (treatment) will be evaluated for sensory attributes, acceptability, and storage stability. After standardizing the techniques for achieving the required fucoxanthin concentrations in the dairy products, at least three replicates of each experiment will be conducted and data collected for statistical analysis.

Progress 01/01/19 to 12/07/21

Outputs
Target Audience:We were planning to present the results of this project at different phases of completion to other food scientists in the professional society meetings of the American Dairy Science Association and the Institute of Food Technology in June and July of 2019 and 2020. However, due to the emergence of pandemic the scheduled plans were disrupted. A part of the results of this project was presented to the annual meeting of the Institute of Food Technologists. Changes/Problems:So far, we have not encountered any major problem that will require major change(s) in the planned approaches for the accomplishments of this project. At the initiation stages of this project, it was a challenge to find food grade fucoxanthin and it took three to four months to locate a reliable source. Finally, a reliable source of food grade fucoxanthin was found from China. What opportunities for training and professional development has the project provided?A post-doctoral fellow (Dr. Maryuri Nunez De Gonzalez) is conducting some of the laboratory experiments on goat milk and yogurt samples that have been supplemented with fucoxanthin. The post-doctoral fellow has been assigned at 50% on this project and has gained considerable experience in food bio-functional ingredients. Additionally, three undergraduate students were assigned to this project during the three-year duration of this project. The undergraduate students that were working on this project presented parts of the data to the Undergraduate Student's Presentations. This project provided unique professional development opportunities for these technical personnel. How have the results been disseminated to communities of interest?A part of the results of this project was prepared as an abstract for oral presentation at the American Dairy Science Association in June of 2020. However, due to the emergence of pandemic the meeting took place in a virtual format, and we postponed the presentation of this abstract. The tittle of the presentation was "Recovery of fucoxanthin in pasturized goat milk"by Rahmat Attaie*, Maryuri Nuñez de González, Selamawit Woldesenbet, Adela Mora-Gutierrez,Yoonsung Jung. Another presentation was prepared for IFT annual meeting in July 0f 2020, using fucoxanthin as an emulsion in food preservation. The title of the presentation was "Chemical stability of fucoxanthin in lipid emulsion: effect of antioxidants". This presentation was also postponed due to the pandemic. However, this abstract was presented virtually in IFT annual meeting in July of 2021. The following paper was published in the peer-reviewed journal of Foods, 2020, 9, 1394; doi:10.3390/foods9101394. The title of the article is "Stability of fucoxanthin in pasteurized skim and whole goat milk". Currently, we are preparing a second manuscript from the results of this project and the title of manuscript is "Physicochemical aspects of goat milk yogurt containing fucoxanthin". The manuscript is in the final stages of preparation and will be submitted to the American Dairy Science Journal shortly. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We were able to accurately measure the content of fucoxanthin in goat milk right after production and throughout its shelf-life under the refrigerated conditions using a HPLC technique. Fucoxanthin was 100% stable and did not undergo degradation during storage at 4°C for four weeks. Additionally, the heat stability of fucoxanthin in goat milk was determined under two different pasteurization temperatures (low-temperature long-time, 64°C for 30min. and high-temperature short-time, 74°C for 15 sec.). Fucoxanthin was very heat stable under these pasteurization conditions and it's percent recovery was nearly 100% as it was measured using a HPLC technique. Fucoxanthin was fully stable and was not destroyed during the heat of pasteurization, which makes fucoxanthin a useful bio-functional supplement in all dairy products. Since most dairy products are pasteurized before consumption, hence heat resistance of fucoxanthin is a useful and necessary property of this bio-functional ingredient. Additionally, the composition of goat milk samples with and without supplementation of fucoxanthin at the initial and final stages of shelf-life were determined using a Lactic check instrument. The density and freezing point of milk samples and the percentages of fat, solid-not-fat, water, and protein were determined. These parameters showed no significant differences between samples containing fucoxanthin and the control milk samples without fucoxanthin. However, the colorimetric analyses of goat milk samples with and without fucoxanthin showed significant differences in the color of milks. This was due to the fact that fucoxanthin is a carotenoid with natural yellow pigments and contributed to the yellowish color of supplemented goat milk. However, due to the coronavirus pandemic a large-scale sensory analysis of this product was not possible. A small-scale study of the organoleptic characteristics of this product was performed in the lab and goat milk with fucoxanthin was acceptable without any objectionable flavor or appearance. Additionally, we developed acceptable quality goat milk yogurt containing fucoxanthin as a supplement. The concentration of fucoxanthin in yogurts were either 5 mg/serving or 8 mg/serving. These products have longer shelf-lives (3 months at 4°C) that can help in better distribution and utilization for human consumption. The physicochemical characteristics, sensory and acceptability of goat milk yogurts supplemented with fucoxanthin were assessed by a small-scale study. All the physicochemical characteristics of yogurts were similar to commercial yogurts and had acceptable sensory attributes. The anti-obesity effect of these goat milk products on human subjects will be the objective of our next research project.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: A part of the results of this project was prepared as an abstract for oral presentation at the American Dairy Science Association in June of 2020. However, due to the emergence of pandemic the meeting took place in a virtual format, and we postponed the presentation of this abstract. The tittle of the presentation was "Recovery of fucoxanthin in pasteurized goat milk" by Rahmat Attaie*, Maryuri Nu�ez de Gonz�lez, Selamawit Woldesenbet, Adela Mora-Gutierrez, Yoonsung Jung. Another presentation was prepared for IFT annual meeting in July 0f 2020, using fucoxanthin as an emulsion in food preservation. The title of the presentation was "Chemical stability of fucoxanthin in lipid emulsion: effect of antioxidants". This presentation was also postponed due to the pandemic. However, this abstract was presented virtually in IFT annual meeting in July of 2021. The following paper was published in the peer-reviewed journal of Foods, 2020, 9, 1394; doi:10.3390/foods9101394. The title of the article is Stability of fucoxanthin in pasteurized skim and whole goat milk".
• Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: Currently, we are preparing a second manuscript from the results of this project and the title of manuscript is Physicochemical aspects of goat milk yogurt containing fucoxanthin. The manuscript is in the final stages of preparation and will be submitted to the American Dairy Science Journal shortly.

Progress 01/01/20 to 12/31/20

Outputs
Target Audience:We were planning to present the results of the initial phases of this project to other food scientists in the professinonal society meetings of the American Dairy Science Association and the Institute of Food Technologists in June and July of 2020. However, due to the emergence of pandemic the plans were postponed and can possibly be presented next year. Changes/Problems:So far we have not encountered any major problems that will require major changes in the planned approaches for the accomplishment of this project. What opportunities for training and professional development has the project provided?A post-doctoral fellow (Dr. Maryuri Nunez De Gonzalez) is conducting some of the laboratory experiments on goat milk and yogurt samples that have been supplemented with Fucoxanthin. The post-doctoral fellow has been assigned at 50% on this project. Additionally, an undergraduate student has been assigned to this project and is currently doing research for undergraduate student's presentations. This project has provided a unique professional development opportunity for these technical personnel. How have the results been disseminated to communities of interest?A part of the results of this project was prepared as an abstract for oral presentation at the American Dairy Science Association in June. However, due to the emergence of pandemic the meeting took place in a virtual format and we postponed the presentation of this abstract possibly for next year. The tittle of the presentation is "Stability of fucoxanthin in pasteurized skim and whole goat milk" by Rahmat Attaie*, Maryuri Nuñez de González, Selamawit Woldesenbet, Adela Mora-Gutierrez, Yoonsung Jung, and Deland Myers. Another presentation was prepared for IFT annual meeting in July using fucoxanthin as an emulsion in food preservation. The title of the presentation is "Chemical stability of fucoxanthin in lipid emulsion: effect of anthioxidants". This presentation was also postponed due to the pandemic and will be presented next year, if conditions allow. A manuscript was prepared and was submitted to the American Journal of Dairy Science, and its title is "Stability of Fucoxanthin in Pasteurized Skim and Whole Goat Milk". The manuscript is in the process. What do you plan to do during the next reporting period to accomplish the goals?Currently, we are working on developing acceptable quality goat milk yogurt with fucoxanthin. The concentration of fucoxanthin in yogurt will be at least 2.4 mg/serving size. This product will have a longer shelf-life (3 months at 4°C) that will help in better distribution and utilization for human consumption.

Impacts
What was accomplished under these goals? We were able to accurately measure the content of fucoxanthin in goat milk right after production and throughout its shelf-life under the refrigerated conditions using a HPLC technique. Fucoxanthin was 100% stable and did not undergo degradation during storage at 4°C for four weeks. Additionally, the heat stability of fucoxanthin in goat milk was determined under two different pasteurization temperatures (low-temperature long-time, 64°C for 30min. and high-temperature short-time, 74°C for 15 sec.). Fucoxanthin was very heat stable under these pasteurization conditions and its percent recovery was nearly 100% as it was measured using a HPLC technique. Fucoxanthin was fully stable and was not destroyed during the heat of pasteurization, which makes fucoxanthin a useful biofunctional supplement in all dairy products. Since most dairy products are pasteurized before consumption, hence heat resistance of fucoxanthin is a useful and necessary property of this supplement. Additionally, the composition of goat milk samples with and without supplementation of fucoxanthin at the initial and final stages of shelf-life were determined using a Lactic check instrument. The density and freezing point of milk samples and the percentages of fat, solid-not-fat, water, and protein were determined. These parameters show no significant differences between samples containing fucoxanthin and the control milk samples without fucoxanthin. However, the colorometric analyses of goat milk samples with and without fucoxanthin showed significant differences in the color of milks. However, due to coronavirus pandemic a large scale sensory analysis of this product was not possible. A small scale study of the organoleptic characteristics of this product was run in the lab and goat milk with fucoxanthin was acceptable without any objectionable flavor or appearance. Currently, we are working on developing acceptable quality goat milk yogurt with fucoxanthin. The concentration of fucoxanthin in yogurt will be at least 2.4 mg/serving size. This product will have a longer shelf-life (3 months at 4°C) that will help in better distribution and utilization for human consumption.

Publications

• Type: Journal Articles Status: Submitted Year Published: 2021 Citation: A manuscript was prepared and was submitted to the American Journal of Dairy Science, and its title is "Stability of Fucoxanthin in Pasteurized Skim and Whole Goat Milk". The manuscript is in the process.

Progress 01/01/19 to 12/31/19

Outputs
Target Audience:We will be presenting the results of the initial phase of this project to other scientists in the professinonal society meetings of the American Dairy Science Association and the Institute of Food Technologists in June and July of 2020. Changes/Problems:So far we have not encountered any major problems that will require major changes in the planned approaches for the accomplishment of this project. What opportunities for training and professional development has the project provided?A post-doctoral fellow (Dr. Maryuri Nunez De Gonzalez) is conducting some of the laboratory experiments and currently has been assigned at 50% on this project. Additionally, an undergraduate student has been assigned to this project and is currently undergoing training. This project has provided a unique professional development opportunity for these students. How have the results been disseminated to communities of interest?The results of this project have not been disseminated to the scientific community yet; however, we are planning to present our data at the annual meetings of the American Dairy Science Association and the Institute of Food Technologists in June and July of 2020. What do you plan to do during the next reporting period to accomplish the goals?We will be adding the required amounts of Fucoxanthin to different milks (whole milk and skim milk) and after processing, run sensory analyses with human taste panels. The overall quality of fucoxanthin containing processed goat milk will be evaluated. Then we will move towards production and dissimination of focuxanthin containing goat milk. The next phase of this project is to manufacture cheese from milk containing Fucoxanthin and hence make acceptable quality cheeses containing sufficient quantity of fucoxanthin as a biofucntioanal ingredient. The composition and overall sensory and physicochemical qualities of cheeses will be assessed and the results will be reported at the scientific meetings and to the food industry.

Impacts
What was accomplished under these goals? We were able to successfully measure the storage stability of fucoxanthin under the refrigerated conditions in goat milk using an HPLC method during 2019. Fucoxanthin was 100% stable and did not undergo degradation during storage. Additionally, heat stability of fucoxanthin was determined by an HPLC method after pasteurization of goat milk and its percent recovery was measured during 2019. Fucoxanthin was fully stable and was not destroyed during heat of pasteurization (65 C for 30 min.) in milk, which makes Fucoxanthin a useful supplement in all dairy products. Since most dairy products are pasteurized before consumption, hence heat resistance of Fucoxanthin is a useful property of this supplement. Additionally, composition of goat milk samples at the initial and final stages of storage were determined using a Lactic check instrument. The density and freezing point of milk samples and percentages of fat, solid-not-fat, water, and protein were determined. These parameters show no significant differences between samples containing Fucoxanthin and the control milk samples without Fucoxanthin. However, the colorometric analysis of goat milk samples with and without Fucoxanthin showed significant differences. The next step is to test the acceptability quality of goat milk samples containing fucoxanthin using human taste panels.

Publications