Source: FORT VALLEY STATE UNIVERSITY submitted to NRP
STRENGTHENING ANIMAL SCIENCE UNDERGRADUATE AND GRADUATE INSTRUCTION BY UPGRADING THE LABORATORIES TO TEACH ANIMAL BIOTECHNOLOGY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
OTHER GRANTS
Reporting Frequency
Annual
Accession No.
0226496
Grant No.
2011-38821-30910
Cumulative Award Amt.
$149,891.00
Proposal No.
2011-02513
Multistate No.
(N/A)
Project Start Date
Sep 1, 2011
Project End Date
Aug 31, 2015
Grant Year
2011
Program Code
[EP]- Teaching Project
Recipient Organization
FORT VALLEY STATE UNIVERSITY
1005 STATE UNIVERSITY DRIVE
FORT VALLEY,GA 31030
Performing Department
ANIMAL SCIENCE
Non Technical Summary
The cloning of animals and sequencing of their entire genomes during the past decade have opened enormous possibilities to enhance the quality and quantity of milk and meat; production of therapeutic proteins in milk of dairy animals; and the use of animals as models of human diseases. These developments, however, require updating the animal science curricula by incorporating molecular technologies to train the next generation of scientists. For effective teaching of molecular technologies adequate hands-on laboratory practices are essential. The overall goal of this project is to enhance the quality of animal science undergraduate and graduate degree programs by incorporating hands-on experience in molecular technologies in instruction. An additional goal is to expose K12 students to agricultural biotechnology through mini-projects and workshops. Such students with practical experience in molecular technology will be better prepared for a) making career decisions, b) pursuing further graduate studies and, c) seeking employment in industry, academia and government agencies.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
100%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
90360993020100%
Knowledge Area
903 - Communication, Education, and Information Delivery;

Subject Of Investigation
6099 - People and communities, general/other;

Field Of Science
3020 - Education;
Goals / Objectives
The specific objectives of this project are, a) to procure, install, and maintain state-of-the-art laboratory equipment, b) to prepare a laboratory manual to teach Animal Biotechnology, c) to enhance student learning of cell and molecular concepts by hands-on laboratory exercises, d) to provide experiential-learning opportunity by conducting research projects for undergraduate, graduate, and K12 students and, e) to conduct workshops to K-12 students and teachers to recruit students in animal science program.
Project Methods
After initial meeting of the project team to discuss the logistics of accomplishing project goals the students will be recruited and the state-of-the-art equipments purchased, and installed to upgrade the lab facilities. Simultaneously, literature will be collected, reviewed and the protocols/lab exercises for undergraduate Animal Biotechnology laboratory course will be written and tested with the help of graduate student and technician. Protocols will be revised based on the class feedbacks/relevance during the course of this project. The undergraduate animal biotechnology laboratory class will be taught using the developed manual and the acquired equipments. Each student will conduct experiment individually as well as in groups (depending upon exercise), record the observations, and will write a lab report for each experiment conducted in the lab. Each lab class will be tied up with theory class for specific topic/concept prior to the lab class. Pre- and post-lab quizzes will be used to measure the student understanding of exercises/concepts after hands-on experiments. The data will be analyzed for statistical significance by using the GraphPad Prism (GraphPad Software Inc., LaJolla, CA). In this project 2 undergraduates and one graduate student will be recruited and directly engaged in research in the area of Animal Biotechnology under the faculty supervision. Each student will prepare a written report of his/her research project and present to faculty. In addition to graduate and undergraduates, K-12 students will also be provided opportunity to conduct mini-research projects in our labs. Workshops will also be arranged for K-12 school students and teachers about the career options in agricultural biotechnology. Finally report will be submitted to USDA and the research results published in appropriate journals.

Progress 09/01/11 to 08/31/15

Outputs
Target Audience:scientific community, students, teachers, farmers, general public Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?More than 115 students have directly benefitted from the project. The equipment, facilities and the wet lab exercises developed during the course of this project provided opportunity for both undergraduate and graduate students in animal science and biotechnology for hands-on learning through laboratory course wor. It will continue each semester beyond the project period.The project also provided opportunity to graduate, undergraduate, and high& middle school students to pursue supervised research projects in the area of animal biotechnology including some with financial assistance. During the project period 4 students graduated with a MS degree in Animal Science/ Biotechnology with a thesis option and 2 students are expected to graduate this year. Students got opportunity to interact and learn from each other in the lab and in weekly lab meetings. They were trained in planning experiments, observing, analyzing data, and writing their project reportsand thesis and developing their presentation posters or lectures. Almost all students presented their work, some of which were provided with opportunity to present even in national/international conferences to a wider audience. They got opportunity to network with other scientists and peers, exposed to various areas of science, and explore career opportunities. Students were closely mentored by PI. Thegraduate and undergraduate students were involved in workshops for high school students and visitors and thus presented withthe opportunity to develop communication skills. How have the results been disseminated to communities of interest?Research results were disseminated to people by presentations in national and international conferences, departmental seminars, agricultural fairs, and through lab orientations to scientists, community visitors, students from different high schools, and through structured workshops on campus and at neighboring high schools. Research results were also published in peer reviewed journals, and on university website. Abstracts were published in conference proceedings. PI also gave invited talks on the educational research activities at FVSU. Impact statement was also submitted to ARD website earlier. What do you plan to do during the next reporting period to accomplish the goals?All the goals of the proposed projecthave beenaccomplished. PI is further submitting funding proposals so as to continue utilization of the equipment, facilities and linkages established from this USDA-NIFA support, and to educate undergraduate and graduate students, and provide opportunities for high school students to conduct mini-research projects in the PI laboratories.

Impacts
What was accomplished under these goals? Objective a: Once the project was approved, PI organized an initial project meeting and discussed the logistics / plans for project implementation with collaborating partners. Equipment quotations were obtained, evaluated, processed, and finally, five sets of pipetting devices, gel tanks, a PCR machine, an automatic cell counter, and a fluorescence microscope were acquired and installed. These equipments are being maintained and used both for teaching wet lab portions of two courses and research in our animal science and biotechnology programs. Since the start of the project in Sep 2011, more than 115 graduate, undergraduate and high& middle school students have benefitted through hands-on labs, thesis/mini-research projects, and/or structured agricultural biotechnology workshops. Objective b: Several wet laboratory exercises have been written and optimized as a part of lab classes to teach ANSC2813-animal biotechnology and ANSC6303-molecular biology techniques courses and, are being taught to undergraduates and graduate students in each fall and spring semesters, respectively. This will continue benefiting future cohort of students. Objective c: To enhance student learning of biotechnological concepts, ANSC2813 course was taught in every fall semester for 3 years by supplementing the lectures with hands-on wet laboratory exercises. Effects of wet lab exercises on learning of biotechnology concepts aws evaluated by pre-post tests. Average pre- and post-test scores of students evaluated on selected exercises showed 14.3 % increase in student learning. Objective d: To provide experiential learning opportunity, the students were involved in supervised research projects. Fourteen students have completed research projects since the initiation of the project in 2011 (4 MS level graduate thesis; 4 BS level; and 6 high & middle school level research projects). Four students have graduated (and 2 are expected to graduate soon) with a MS degree in animal science/ biotechnology with research thesis option. All students presented and defended their thesis proposal to animal science and biotechnology faculty. Three students including one undergraduate student presented their research results in ARD symposium in Jacksonville, FL in 2013, and one student presented his research to international audiences in the Society for In Vitro Biology meeting at Savannah in June 2014. Five students presented in county and/or state level science fairs, and almost all of them won some or the other award in science fairs. Research work of 2 graduate students has already been published in peer reviewed journals, and 3 other manuscripts are being developed. In all 4 peer-reviewed papers and11 conference abstracts have been published. Project scientists have given 3 invited lectures and several other presentations. One significant achievement of student research results was that it demonstrated that the individual cells in postmortem mammalian tissues remain alive for much longer time than was previously thought. It has significant implications in transplantation biology and preservation of superior genetics for cloning of animals to meet the climatic and other disastersin future. We have also developed methodologies to procure and preserve viable cells from postmortem animal tissues. We have developed, characterized and successfully cryopreserved several goat and sheep fetal and adult fibroblast cell lines that will be used for genetic manipulation of goats and sheep in future research projects. These facilities for tissue and cell banking will soon be made available to Georgia farmers. Objective e: To expose K12 students to agricultural biotechnology, we conducted two workshops. A whole day workshop that involved lectures, wet labs, videos, and paper modeling was conducted by project faculty and students in June 2012. It involved15 high school students selected in Ag Discovery program at FVSU. Pre/post-evaluation tests indicated a significant learning pattern by these students. Another workshop was conducted to about 30 high school students visiting from Mount De Sales Academy, Macon in 2014.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Okonkwo, C; and M. Singh. Recovery of fibroblast-like cells from refrigerated goat skin up to 41 days of animal death. In Vitro Cell and Developmental Biology-Animal. (2015) 51: 463-69 (DOI 10.1007/s11626-014-9856-9)
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Singh, M; and X. Ma. In Vitro Culture of Fibroblast-Like Cells From Sheep Ear Skin Stored at 25-26�C for 10 Days After Animal Death. International Journal of Biology (2014) 6 (4): 96-102 (doi:10.5539/ijb.v6n4p96)
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Adefenwa, MA, B O Agaviezor, SO Peters, M Wheto, K Adekoya, M Okpeku, B Oboh, GO Williams, M Singh and IG Imumorin. Identification of ingle nucleotide polymorphisms in the agouti signaling protein (ASIP) gene in Nigerian and North American goats. Molecular Biology Reports (2013) 40(7): 4447-4457 (doi: 10.1007/s11033-013-2535-1)
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Singh, M; X. Ma & A. Sharma. Effect of postmortem time interval on in vitro culture potential of goat skin tissues stored at room temperature. In Vitro Cell and Developmental Biology-Animal. (2012) 48: 478-482 (doi: 10.1007/s11626-012-9539-3)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Aoued, H.S., M. Singh. Recovery of fibroblast-like cells after 160 days of postmortem storage of goat skin tissues in refrigerated media. In Vitro Cell. Dev. Biol.-Animal (2014) 50: S40. DOI 10.1007/s11626-014-9765-y (Abstr.).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Singh, M; X. Ma, G. Kannan, and E. Amoah. Time limits of postmortem cell survival in goat ear skin stored at room temperature. J. Animal Sci. (2013) 91 (E. Suppl.): pp220 (Abstr).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Okonkwo, C; X. Ma and M. Singh. Postmortem Cell Survival Pattern of Goat Skin Tissues Stored in Refrigerator at 4 degree C. 17th Biennial ARD Research symposium, Gainesville, FL April 7-10, 2013, Abstract No: 11, page 50.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: SINGH, M; X. Ma, C. Okonkwo, N. Degala, P. Yadav , O. White, E. Amoah, W. Getz, and C. Borne. Enhancing student learning with hands-on laboratory exercises and research projects in animal biotechnology. NACTA Journal, Vol. 57, supplement 1 (2013) pp 96-97, abstract no 132.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Singh, M; and X. Ma. Post-mortem Recovery of Fibroblast-like Cells in Sheep after 10 Days of Animal Death. In Vitro Cell. Dev. Biol.-Animal (2013) 49: S40-46. DOI: 10.1007/S11626-013-9605-5. (Abstr.).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Singh, M; and X. Ma. Sheep Skin Fibroblast Cell Cultures from Room Temperature Stored Tissues after Ten Days of Animal Death. 17th Biennial ARD Research symposium, Gainesville, FL April 7-10, 2013, Abstract No: 206. Page 120.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: White, O; X. Ma, and M. Singh. Evaluation of Fetal Goat Serum as a Cheaper Alternative to Fetal Bovine Serum for Culturing Goat Cells. 17th Biennial ARD Research symposium, Gainesville, FL April 7-10, 2013, Abstract No: 31, Page 57.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Degala, V.N.; X. Ma, E. Amoah, and M. Singh. Development of a Cell Biological Tool for Genetic Modification of Goats. 17th Biennial ARD Research symposium, Gainesville, FL April 7-10, 2013, Abstract No: 319, page 162.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Singh, M; X. Ma and A. K. Sharma. In vitro culture of fibroblast-like cells from room temperature stored goat tissue. In Vitro Cell. Dev. Biol.-Animal (2012) 48: S50-51. (Abstr.).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Singh, M; and X. Ma. Ability to culture of cells from postmortem goat skin tissues stored at room temperature for different time intervals. J. Animal Sci. (2012) 90 (Suppl.3): 674 (Abstr).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Singh, M. Can dead animals be revived after several days of their death? Oral presentation given in 2nd FVSU Research Day, April 12, 2012. Abstract No: O7, p34
  • Type: Other Status: Other Year Published: 2015 Citation: Singh, M. (2015). How long individual cells are alive in postmortem mammalian tissues? Lecture delivered at National Dairy Research Institute, Karnal, India; March 16, 2015.
  • Type: Other Status: Other Year Published: 2015 Citation: Singh, M. (2015). Postmortem cell survival in mammals: potential for future cloning of animals. Lecture delivered at Institute of Himalayan Bioresource Technology, Palampur, India; March 19, 2015.
  • Type: Theses/Dissertations Status: Published Year Published: 2013 Citation: Charles Okonkwo (Fall 2013). Studies on postmortem survival of goat skin tissues stored in a refrigerator, MS thesis submitted to Fort Valley State University.
  • Type: Theses/Dissertations Status: Awaiting Publication Year Published: 2014 Citation: Nagender Degala (Summer 2014). Establishment and characterization of fetal fibroblast cell lines in Capra hircus (goats); MS thesis submitted to Fort Valley State University.
  • Type: Theses/Dissertations Status: Awaiting Publication Year Published: 2015 Citation: Olabisi Ojo (Spring 2015). Establisment of goat fetal and adult fibroblast cell lines and their comparative freezing cell viability and transfection efficiencies. MS thesis submitted to Fort Valley State University.
  • Type: Theses/Dissertations Status: Awaiting Publication Year Published: 2015 Citation: Hadj S. Aoued (Spring 2015). Studies on postmortem tissue survival in goats: effect of storage in refrigerated media; MS thesis submitted to Fort Valley State University.


Progress 09/01/12 to 08/31/13

Outputs
Target Audience: students (high and middle school; undergraduate and graduate students); scientists, general public including parents Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? During this reporting periodthe project provided opportunity for 2 graduates, 1 undergraduate and 1 high school students to conduct research in the lab. Students got opportunity to lear biotechthrough workshops, classroomsemesterwise lab exercisesin ANSC2813 animal biotechnology class offered in fall, and molecular biology techniques lab class. One of graduate syudents has submitted his MS thesis and has graduated in MS animal Science in fall 2013. How have the results been disseminated to communities of interest? The results of the study were presented in society meetings and in ARD conference by PI and students. Publications infoormationisalso uplodaded onfaculty website.Opportunitiesin agriculturalbiotech are also discussed routinely to visitors and in openhouses to recruit students What do you plan to do during the next reporting period to accomplish the goals? Plan to teach ANSC2813 class in fall semester to re-evaluate student learning by hands on lab exercises. Also plan to complete remaining experiments anddata analysis ofstudent research. Plan to revise laboratory exercises based on fall ANSC2813 class experience and for use in subsequent years.

Impacts
What was accomplished under these goals? Goal a): equipments were procured, installed and are being used in labs (reported earlier); goal b&c): 2 more lab exercises were written and ANSC2813-animal biotechnology course was taught in fall 2012 by supplementing the lectures with hands-on laboratory exercises. Pre- and post-test scores of students evaluated on selected exercises showed an average difference of 33.1% for the undergraduate class. goal d): this project period graduate committee was formed for one graduate student who was recruitedin previousyear. The student developed and presented his MS thesis proposal to animal science and biotechnology faculty. He subsequently conducted research to evaluatethe time limits of postmortemcell survival in goat skin tissues stored in refrigerator in order to recover cells that can be cultured in vitro and cryopreserved for future cloning of preserved animals. The student presented research resultsin ARD symposium in Jacksonville, FL. An undergraduate student who was trained last year in cell and molecular biology techniques was recruited again and motivated to conduct research resulting in a presentation in ARD symposium. The goal of the student project was to evaluate, if the fetal goat’s serum (FGS) which is freely available at the university slaughter house can be used as a cheaper alternative to fetal bovine serum to culture mammalian cells. GSF-289 cell line that was earlier developed in our lab was used to test the effect of goat serum on growth. The proliferation rate of cells with different serum supplementation was observed both qualitatively (under inverted microscope) and quantitatively (by counting total number of cells) after 3-4 days of cell culture. An 8th grader from Fagan mil middle school also conducted a research project entitled, “Plasmid DNA transformation in E. Coli: effect of microwave on transformation efficiency”. Student received first prize for his project in regional science fair and was selected for Georgia State Science Fair in UGA Athens where also he received two prizes. Goal e): In addition to a workshop to K12 students in last reporting period we conducted a workshop to visiting high school students from Mout De sales academy, macon in this year as well inspiring them to ag biotechnology. In addition, PI attended PDs meeting at Virginia tech and presented a paper in NACTA conference.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Okonkwo, C*; X. Ma and M. Singh. Postmortem Cell Survival Pattern of Goat Skin Tissues Stored in Refrigerator at 4 degree C. 17th Biennial ARD Research symposium, Gainesville, FL April 7-10, 2013, Abstract No: 11, page 50.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: White, O*; X. Ma, and M. Singh. Evaluation of Fetal Goat Serum as a Cheaper Alternative to Fetal Bovine Serum for Culturing Goat Cells. 17th Biennial ARD Research symposium, Gainesville, FL April 7-10, 2013, Abstract No: 31, Page 57.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Singh, M*; and X. Ma. Post-mortem Recovery of Fibroblast-like Cells in Sheep after 10 Days of Animal Death. In Vitro Cell. Dev. Biol.-Animal (2013) 49: S40-46. DOI: 10.1007/S11626-013-9605-5. (Abstr.). Interactive poster presentation no: A2000.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Singh, M*, X. Ma, C. Okonkwo, N. Degala, P. Yadav, O. White, E. Amoah, W. Getz, and C. Borne. Enhancing student learning with hands-on laboratory exercises and research projects in animal biotechnology. NACTA Journal, Vol. 57, supplement 1 (2013) pp 96-97, abstract no 132. Presented in NACTA Conference, at Virginia Institute of Technology, June 25-29, 2013.


Progress 09/01/11 to 08/31/12

Outputs
OUTPUTS: Quality of undergraduate and graduate degree programs was improved by incorporating hands-on laboratory experience by utilizing the equipment and supplies support. After hearing the outcome of the proposal, we began to organize the animal biotechnology lab and aggressively worked to recruit students. Once the project was initiated on September 1, 2011, an initial project meeting was held which was attended by Drs. M. Singh, E. Amoah, Will Getz, C. Borne and Mr. Donovan Coly our Ag marketing personnel. The logistics/ plans for project implementation were discussed and subsequently followed. Enquiries of equipment were made from several venders and quotations were obtained and processed. Finally, five sets of pipetting devices, a real time PCR machine, an automatic cell counter, and a fluorescence microscope were acquired and installed. These equipments are, currently, being maintained and used both for teaching and research in our animal science and biotechnology programs, thus completing objective 1. Soon after the project initiation, one animal science graduate and one animal science undergraduate student was recruited. In fall 2012, one undergraduate biology major and one animal science major were also recruited and are pursuing research projects. These students were initially trained in procuring/processing goat tissues for primary fibroblast cell cultures from adult/fetal tissues and in general cell/molecular biology techniques and are currently pursuing individual research projects in PI's lab. One of the projects undertaken in the lab involved testing postmortem storage limits (at ambient temperature simulating natural farm environment) of goat skin tissues in order to preserve the superior germplasm for future use. To achieve this objective, 2-3 mm square skin pieces (70 nos) from ears of 3 breeds of goats (7 goats) after 0, 2, 4 and 6 days of postmortem storage at 24 degree C were cultured and outgrowing cells observed after 10days. Results showed that outgrowth reduced with increasing time interval. Comparison of the cells from 0- and 6-dpm cultures revealed similar cell morphology, growth curves, and doubling times, suggesting that cultured cells from 6 dpm tissues can be used for preserving superior genetics for future. Two laboratory protocols, a) DNA extraction and, 2) gel electrophoresis were also written, tested in the lab, and are currently being taught in ANSC 2813-Animal Biotechnology and, ANSC 6303-Molecular biology techniques lab course. The project also provided an opportunity to a middle school student to conduct research project entitled, "Effect of magnesium ion concentration on PCR amplification of myostain gene in goats." He presented his research in Houston County Science and Engineering Fair (Feb. 2012), and received first prize for his project. In addition, a whole day workshop on DNA extraction and gel electrophoresis was also conducted to 15 high school students in June 2012. It involved lectures, wet labs, videos and paper modeling. Pre/post-evaluation tests indicated a significant learning pattern by students. PARTICIPANTS: Mahipal Singh, Xiaoling Ma TARGET AUDIENCES: scientific community, students PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The facilities, supplies and stipend that could become possible from this project fund have been and are being utilized to provide hands on experience to K-12, undergraduate and graduate students through laboratory course work (16 students), day long workshops (15 students), and through independent student mini-research projects (5 students). Laboratory environment provided an opportunity for graduate, undergraduate and K-12 students to interact among themselves and with faculty and technical staff. It has created an excellent environment for developing interpersonal and scientific communication skills along with technical competence. The hands on laboratory based education in animal science discipline will make minority students competitive for higher education opportunities and in tomorrow's knowledge workforce. Additionally, the results of our research experiments on postmortem tissue survival at ambient temperatures suggest that live cells can be recovered from goats and perhaps other animals even after 6 days of their death with comparable growth profiles and thus can be used for tissue banking for preservation of superior genetics, genetic diversity and cloning of animals.

Publications

  • Singh, et.al. (2012). Effect of postmortem time interval on in vitro culture potential of goat skin tissues stored at room temperature. In Vitro Cell and Developmental Biology-Animal. 48: online first (DOI: 10.1007/s11626-012-9539-3)
  • Singh, et.al. (2012). Ability to culture of cells from postmortem goat skin tissues stored at room temperature for different time intervals. Journal of Animal Science 90 (Suppl.3): 674 (Abstr). (Oral presentation)
  • Singh, M. (2012). Can dead animals be revived after several days of their death FVSU Research Day Symposium, April 12, 2012. Abstract No: O7, p34 (Oral presentation)