Institutional Animal Use and Care Committee

Transcription

1 Copies: Office of Scholarship and Sponsored Projects IACUC Chair Principal Investigator Institutional Animal Use and Care Committee Research and Scholarly Activities Protocol Application Form This form is to be completed by Principal Investigators, graduate students, curators or Museum Directors. When the form is complete, submit using the Submit form button in the upper right corner. Appropriate and current licensing/collection permits must be on file in the Office of Scholarship and Sponsored Programs, 306D Picken Hall or attached to the Submit Form . Please keep in mind that: 1. This application is designed primarily for studies that involve warm- - - blooded animals (mammals and birds) conducted at FHSU in compliance with the USDA Animal Welfare Act. However, all activities that involve use of ANY vertebrate species (fish, amphibians, reptiles) at FHSU must have completed and approved protocol forms before the activities can be initiated. 2. The protocol must be approved prior to the initiation of project/sample collection. 3. Protocol reviews are scheduled for twice a semester. Be sure that the protocol application is complete and submitted in a timely fashion in order to avoid unnecessary delay. 4. When receiving funding through intramural sources (URE, K- - - INBRE, etc.), the protocol must be submitted for the review prior to submitting the grant application to the respective offices, and the approval number must be included in the grant application. 5. Housing of animals, particularly warm- - - blooded animals (mammals and birds), is restricted and limited. The principal investigator must receive approval for the use of an animal housing facility, including agreement on per diem of housing throughout the duration of the project, from the academic unit chair prior to submission of this form. 6. FHSU does NOT house a veterinarian on- - - campus and does NOT have surgical facilities. Therefore, any protocols involving survival surgery is prohibited. If survival surgery is unavoidable, the principal investigator must make appropriate arrangement with a licensed veterinarian prior to submitting the application. 7. FHSU does NOT possess a license for possession of radioactive isotopes that can be used for biological procedures. Therefore, use of radioactive isotopes on animals is prohibited. 8. Persons working with animals need to document completion of handling and safety training that is appropriate for the species/group under investigation. Completion of the appropriate online CITI training is required for all personnel listed as involved in the activity. 9. The use of hazardous materials requires documentation of authorization from the university environmental safety supervisor, Bill Deeds. 10. This form is also applicable for any instructional activities that contribute to research, collections and exhibits. 5/17/2012 1

2 Section 1: General Information Principal Investigator Name (Last, First, MI):Kobayashi, Yasuhiro (Yass) Degree: Ph.D Title: Assistant Professor of Biology Department, including building and room # Biological Sciences, 410 Albertson E- - - mail: y_kobayashi@fhsu.edu On- - - Campus telephone: Fax: Emergency Contact (Home, mobile): Please list all co- - - investigator(s): Co- - - investigator Department Institution Co- - - investigator Department Institution Co- - - investigator Department Institution Proposed title of the project: Identification of potential genetic markers for detection of estrogeinc endocrine disrupters in non- migratory freshwater fish Proposed project start date: 5/10/2012 Proposed project termination date: 5/31/2015 *Upon approval, the protocol is in effect for 3 (three) years, starting at the proposed start date, regardless of the termination date. The PI must submit a request form for extension PRIOR to the expiration date. Otherwise, a new protocol application must be submitted. Type of protocol (Check one) X New Continuation/Renewal Resubmission Type of project (Check one) X Research Museum collection Exhibition

3 Funding Source Extramural (list the name of funding agency, title of grant, and business office account number): Intramural (Check one): Academic Department (specify) Graduate School URE X K- INBRE Other Section 2: Animal Information General information Type of animal(s) used (check all apply) Warm- blooded animals Mammals Domesticated Non- domesticated Birds Domesticated Non- domesticated Cold- blooded animals Amphibians Captive- reared Wild- caught Reptiles Captive- reared Wild- caught Fish X Captive- reared X Wild- caught If using non- - - domesticated mammals or birds, please complete the following section along with section 2-2. Otherwise, proceed to section 2-2. Does the proposed study require any federal, state, and/or local permits? X Yes No If yes, the PI must attach or have on file with the office of Scholarship and Sponsored Projects the appropriate current permit when using non- - - domesticated mammals or birds, or wild- - - caught cold- - - blooded animals. Do animals used in the proposed study require registration/license from the USDA? Yes X No If yes, please contact the Office of Scholarship and Sponsored Programs.

4 Will the proposed study (including observation of vertebrates in the natural environment) be conducted in accordance with established guideline(s) that is available through the appropriate professional society? Yes X No N/A If yes, please provide the name of the society with the guideline that will be used in the proposed study Specific information Species Strains (if applicable) Source Number anticipated* 300 Walleye Milford hatchery/cider Bluff reservoir Channel Catfish Milford hatchery/local watershed 300 Orangethroat darter Local watershed 100 Northern plains killifish Local watershed 100 *Total number of animals expected to be used during the duration of protocol (3 years) Housing (Check one) University Farms (Domestic cattle, sheep, and swine only) Sternberg Museum (Animals for exhibition and food source only) FHSU Wetlands Educational Center (Animals for exhibition and food source only) FHSU Ecological Resource Center ( Cold- - - blooded animals, temporary holding of warm- - - blooded animals only) X FHSU Greenhouse/105 Facilities and Grounds building (Fish only) No housing facilities needed Briefly describe any additional housing needs in the space below:

5 Section 3: Description of project Overall description and aim Briefly (200 words or less) describe the project in easy to understand language, including its value and scientific significance, and the justification for using animals. The committee is composed of faculty members from non- - - scientific departments, a community member, and university administrators. Therefore, scientific terms and specific laboratory procedures must be written in a manner that can be understood by non- - - scientists and laypersons. Many industrial and household chemicals influence the reproduction of animals negatively because these chemicals mimic the action of estradiol, which is the chief female steroid hormone. Further, native and synthetic estrogens enter the watershed via sewer system and negatively impact animal reproduction. The proposed study is designed to identify potential genetic markers whose expression is influenced by the presence of estrogen and to determine whether their expressions are different based on human activity. For the proposed study, we will first characterize genes associated with neuroendocrine regulation of reproduction (kisspeptin), gonadotropins, and an enzyme that produces estrogen called aromatase, whose expression is shown to be influenced by estrogenic contaminants, in various fish species found commonly in Kansas including walleye, channel catfish, orangethroat darter, and northern plains killifish. We will characterize which tissues express these genes and examine changes in their expression during normal life history of these fish. We will then treat fish with estrogen by either feeding fish with estrogen- traced feed or injecting it into the abdominal cavity. We will harvest the brain and other tissues from treated fish and examine gene expressions. We will also harvest fish from different watershed that are exposed to different degree of human activities as well as different types of human activities and examine expression of these genes. Although the focus of this study is on fish reproduction, this study has a critical implication to humans because contamination of watershed may indicate potential contamination of drinking water. In addition, estrogenic compounds become increasingly accumulated as larger predatory animals consume smaller preys, meaning that large predatory fish such as channel catfish and walleye, which are highly sought- after game fish, accumulate these chemicals, and humans consume these fish are potentially exposed to these chemicals.

6 Assurance of novelty The PI must conduct literature search prior to the submission of the protocol to ensure that there will be no unnecessary duplication of the study. Date(s) of search: 05/01/2011 & 05/21/2012 Database(s) searched: PubMed/Medline Term(s) used for the search: Aromatase, kisspeptin, gonadotropins, catfish, orangethroat darter, northern plains killifish fish, walleye, channel Summary of search results: No publications found in species of interests regarding these topics Justification of animal use Briefly (200 words or less) justify why the use of animals is necessary for the successful completion of the project. We are interested in investigating the effects of environmental contamination/human activities on the reproductive axis of fish from the whole organism perspective, especially within the neuroendocrine axis that controls reproduction. There are no suitable in vitro/ex vivo models available for the studies such as this Non- - - animal model consideration. Briefly explain how the use of non- - - animal model was considered. There are some cell culture system available to detect presence of estrogenic endocrine disrupters in the water. However, we are interested in how human activities impact reproduction of fish in the wild and how changes in expression of these gene markers are associated with reproductive process of fish. Therefore, we must use whole organisms for this study

7 Section 4. Humane Treatment Consideration Pain categories Does the protocol require physical manipulation that could potentially result in more than momentary discomfort to the animals? X Yes No If yes, please check the pain/distress category that best describes the protocol Pain Category B (Bred, Conditioned, or Held for use) Pain Category C (No or Momentary Pain and/or Distress) X Pain Category D (Alleviated Pain and/or Distress) Pain Category E (Unalleviated Pain and/or Distress) If the protocol falls under category D, please describe how pain/distress will be alleviated. Be sure to include the type of anesthesia/analgesia/sedatives used in the study and route of administration. Fish will be anesthetized by using a commercially available anesthesia called Tricaine- S(Tricaine methane sulfonate/ms- 222). The anesthesia will be dissolved in the water in a 5- gallon bucket before the administration, and fish will be anesthetized in the bucket prior to the procedure. If the protocol falls under category E, provide justification why alleviation will not be administered. If the protocol falls under either category D or E, you must provide the name of the supervising veterinarian and his/her contact information in the space below. N/A. Anesthesia is available for commercial use (non- prescription drug) Veterinarian Phone

8 Euthanasia Will the animals be euthanized as a part of the protocol? X Yes No If yes, please provide the method of euthanasia and the person responsible for administration. We will use the same drug used for anesthetizing fish for euthanasia. We will dissolve an overdose of Tricane- S in the water and place fish in the water. Dr. Kobayashi will administer the drug during all procedures. Briefly justify the use of euthanasia We are collecting brain, liver, and gonads from fish. It is inhumane to collect these tissues without euthanasia Disposition of animals after the study* What are the plans for the animals that are either not included in sample collection or those that are included in a non- - - euthanasia sample collection? (Check all applicable) X Euthanasia Adoption Long- - - term retention/holding Transfer to another PI upon protocol approval (provide name) Other (specify) *Any domestic mammals and birds, as well as other animals that are captive- - - reared CANNOT BE RELEASED into the environment/wild population.

9 Use of hazardous materials listed below requires documentation from the university environmental safety supervisor, Bill Deeds. The documentation must be attached to this application. Does the proposed study involve the use of the following: Biological/infectious agent(s)* Yes X No If yes, provide the name(s) of agent(s) and route of administration: Recombinant DNA* Yes X No If yes, list: Hazardous Chemicals Yes X No If yes, list: : Carcinogens/tumor cells Yes X No If yes, list: : Other Yes X No Describe the materials that will be used: Section 5. Personnel and Training For research activities, the PI must document that all personnel involved in the proposed study have received appropriate training in humane handling and care of animals. It is the PI s responsibility that personnel involved in the study must be professionally trained. Personnel Department FHSU e- - - mail address Training Yass Kobayashi Biological Sciences y_kobayashi@fhsu.edu Abigail Pflughoeft Biological Sciences Amber Honig Biological Sciences Cammie Schmitdberger Biological Sciences Briefly describe relevant experience/expertise and training of each person listed above. Dr. Kobayashi has over 30 years working with various animals withexpertise in working with domestic animals (over 25 years) and fish (5 years). Dr. Kobayashi has been formally trained in humane care of animals at the University of Missouri- Columbia, University of Arizona, and Michigan State University. For the last 5 years, Dr. Kobayashi has worked exclusively on fish under the supervision of the USDA scientists. All students listed above are the undergraduate students who are working under direct supervision of Dr. Kobayashi, and they are not allowed to work with animals (with exception of routine daily care) without Dr. Kobayashi being present.

10 Section 6. Consultation List name(s) of FHSU faculty member/outside experts that possess expertise in the animal species and experimental procedure described in this protocol. N/A Section 7. Investigator Assurance I, Yass Kobayashi agree to abide by the USDA Animal Welfare Regulations (CFR 1985). I also have reviewed and agree to abide by all institutional policies governing the use of vertebrate animals for research, testing, teaching, or demonstration purposes at FHSU. I assure that the proposed studies are not unnecessary duplication of experiments conducted previously. I also assure that animals used in the study will receive humane care throughout the duration of the study and that, in the event that animals in this study experience pain or distress which cannot be adequately relieved, animals will be euthanized humanely and immediately based on recommendations of the veterinarian or IACUC. The PI s name should be typed below. Actual signatures will be required following IACUC approval. Principal Investigator IACUC Chair Office of Scholarship and Sponsored Projects Date Date Date The approval period is for three years from the date this protocol is approved. The Principal Investigator is responsible for submitting the appropriate forms to modify the approved activities or for a renewal in a timely manner.