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UST IACUC Animal Care and Use Statement Protocol Review Form
June 2016
University of Santo Tomas Institutional Animal Care and Use Committee ANIMAL CARE AND USE STATEMENT (Protocol Review Form)
UST-IACUC Code: ____________________
Application Fee Received by: _____________ (PRINT Name & Sign)
BAI Certificate No.: ____________________ (For UST-IACUC) ------------------------------------------------------------------------------------------------------------------------Instructions.
(a) Please complete the form by giving all the details asked for. (b) Have the Protocol Review Form SIGNED and DATED by your respective ADVISER/S & DEAN. (c) Submit the accomplished Protocol Review Form to the IACUC Office at the Research Center for the Natural Sciences (Rm. 202 TARC) for review and approval prior the conduct of the scientific work involving animals.
RESPONSIBLE PERSON OR PRINCIPAL INVESTIGATOR [PRINT Complete Name, Student Number & Contact Details (cell phone number and email address)]
Name: Diana Patricia D. Bala Student Number: 2014 - 062605 Email Address:
[email protected]
Cellphone number: 09175627781
Name: Allyssa Jiselle M. Cabalonga Student Number: 2014 - 062663 Email Address:
[email protected]
Cellphone number: 09178808647
Name: Caitlene Joy C. Mejia Student Number: 2014-065613 Email Address:
[email protected]
Cellphone number: 09179701913
NAME OF ADVISER/S &/OR CO-ADVISER/S [PRINT Complete Name, College of Affiliation & Contact Details (cell phone number and email address)]
Adviser: Affiliation:
Email Address: Co-adviser: Affiliation:
Email Address: Cellphone number:
Asst. Prof. Jose Francis V. Abrantes, M.Sc.
Department of Biology College of Science and University of Santo Tomas
[email protected] Prof. Gina R. Dedeles, PhD. Department of Biology College of Science and Research Center for Natural and Applied Sciences University of Santo Tomas
[email protected] 09185710803
R CN A S - UST US T I A CUC CU C Ani mal Care and Use State Statem ment Pr otoco tocoll R eview Form F orm
UST: US T: S037-00-FO01 S037-00-F O01
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UST IACUC Animal Care and Use Statement Protocol Review Form
June 2016
PROCEDURE(S) OR TITLE OF RESEARCH/STUDY: Isolation and Identification of Marine yeasts from Mangrove leaves as feed supplement for Oreochromis niloticus PURPOSE/OBJECTIVES: This study aims to isolate and identify marine yeasts from mangrove leaves, to screen protein content of identified marine yeast, and to evaluate their efficacy as feed supplement to Oreochromis niloticus DURATION OR TIME FRAME: 8 weeks BACKGROUND AND SIGNIFICANCE OF THE PROCEDURE OR RESEARCH: Oreochromis niloticus, also known as the Nile Tilapia, is one of the useful species of cichlids. Its fast growth, firm and tasty flesh, resistance against harsh conditions and ease production of fingerlings under captivity makes it as an important species in aquaculture. The characteristics of O. niloticus make it as an excellent model for studying developmental in cichlids because of its high fecundity, a breeding capability which is 2-3 week spawning cycle, and a short generation period. Marine microorganisms like marine yeasts live in extreme environments, providing a unique potential for the synthesis of functional biomolecules (Connell et al., 2008). A true marine yeast must be able to grow on or in a marine substrate (Kutty & Philip, 2008). Marine yeasts are yeasts that are isolated from marine environments, can grow better on a medium prepared using seawater rather than fresh water (Chi et al., 2010). Since almost all yeasts can grow and tolerate high salinity concentrations, therefore, salinity tolerance does not distinguish marine species from terrestrial species (Kutty & Philip, 2008). According to reports, marine yeasts are able to produce many bioactive substances, such as amino acids, glucans, glutathione, toxins, enzymes, phytase and vitamins with potential application in the food, pharmaceutical, cosmetic and chemical industries as well as for marine culture and environmental protection (Chi et al., 2009; Sarkar et al., 2010). The factors affecting the distribution of marine yeasts include currents, migration of marine organisms, and contamination from terrigenous sources (van Uden & Branco 1963, Fell 1967, Vogel et al. 2007, Kutty & Philip 2008). Marine yeasts are known to be agents of biodegradation. Utilization of this potential in the biotransformation of cheap raw materials or waste matter into value-added products will be a highly rewarding endeavor. Biotransformation and the partial conversion of raw material into yeast biomass are very significant in the nutritional composition of yeast and its potential use as animal or aquaculture feed (Sarlin, P. J., 2005) Marine yeasts are commonly found growing on mangrove leaves and on bodies of water. These organisms are not well studied in the Philippines and thus this research could contribute to knowing more about the organisms. The findings of the study can have a great contribution to the fishing industry. This study can help achieve the optimum growth of Nile tilapias and decrease its mortality rate. It could be cost-effective, helps lessen the use of commercially produced feeds and use natural products as an alternative. DESCRIPTION OF METHODOLOGIES/EXPERIMENTAL DESIGN: A. Type of animal to be used: Juvenile Tilapias ( Oreochromis niloticus) B. Source of the animals: University of the Philippines National Institute of Health Building, 623 Pedro Gil Street,
RCN AS - UST I ACUC Ani mal Care and Use Statement Pr otocol R eview Form
UST: S037-00-FO01
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UST IACUC Animal Care and Use Statement Protocol Review Form
June 2016
Ermita, Manila, 1000 Metro Manila C. Reason/Basis for selecting the animal species: Model animal for polyclonal antibody production (Hanly et al., 1995)
D. Sex and number of animals:
Sex/Age: Male or Female/1-3 days old Number of animals per treatment:
10
Total number of animals:
50
E. Quarantine and/or acclimation or conditioning process
Indicate where: Animal house, UST Thomas Aquinas Research Complex (5 th floor)
Duration: 60 days
Conditions F. Animal care procedures
1. Cage Type & Size: 30” X 36” X 42” steel cages (elevated) none used Type of Bedding: 2. Number of animals per cage: 10 3. Cage cleaning method: Aquarium is equipped with air pump and filter. The water will be changed every week. 4. Room temperature: 25.5 ± 1.4 °C 7.9 ± 0.2 pH: Humidity: 50% humidity Ventilation: A clean, dirt and smoke free environment with fans for the rabbits Lighting: 12 hours light and 12 hours dark 5. Animal diet and feeding Diet concentrations: 100% feeds 80% feeds 20% marine yeasts 70% feeds 30% marine yeasts 60% feeds 40% marine yeasts 50% feeds 50% marine yeasts Feed: Muesli Rabbit Feeds (BioResearch) Water: Purified Drinking Water (Absolute Distilled Drinking Water, Asia Brewery, Inc.) G. Experimental or animal manipulation methods The protocol that will be used will be based on Thermo Fisher Scientific’s Custom Rabbit Polyclonal Antibody Production Protocols (Thermo Fisher Scientific, n.d. ) 1. General description of animal manipulation methods Nile Tilapias will be used to measure the efficacy of marine yeasts as a feed supplement. Different diets will be fed to the tilapias, and thei r body weight and length will be measured after 60 days
RCN AS - UST I ACUC Ani mal Care and Use Statement Pr otocol R eview Form
UST: S037-00-FO01
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UST IACUC Animal Care and Use Statement Protocol Review Form
June 2016
2. Dosing method: Antigen shot: Two (2) ml of the synthetic peptide admixed with 2 mL of complete/incomplete adjuvant. The initial antigen shot (2nd week) will make use of a complete adjuvant preparation while the booster doses (4th and 6th week) will use the incomplete adjuvant preparation (refer to table below). Two hundred μl of the antigen will be subcutaneously injected in 10 sites (dorsolumbar to the ventrothoracic and abdominal regions; refer to figure below). 3. Specimen or biological agent: Blood will be collected through the auricular marginal vein using a 5 ml 21-gauge needle syringe. Three (3) ml of blood will be collected once every week except for the final (8th) week where 5-10 ml of blood will be collected. Week Antigen Shot
1
2
3
✔
Blood Collection
✔
✔
4
5
✔ ✔
✔
6
7
8
✔ ✔
✔
✔
✔
Diagram of a rabbit showing the injection sites [red spots; on both left and right sides (total of 10)]. 4. Animal examination procedures and frequency of examinations: animals will be examined daily by the researcher. 1/2 cup of pellets per 6 lbs. of body weight daily Drinking water must be constantly supplied Check for cages and lock stability Regular check of humidity and temperature of the animal house
5. Use of anesthetics: none. i. ii. iii.
Where will this be bought? NA Who will administer? NA Licensed veterinarian: Name & License no.
NA
6. Surgical procedures: none. 7. Euthanasia: none.
RCN AS - UST I ACUC Ani mal Care and Use Statement Pr otocol R eview Form
UST: S037-00-FO01
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UST IACUC Animal Care and Use Statement Protocol Review Form
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8. Animal Disposal: Veterinarian will be informed of any dead animal that has been used in the experiment. Dead animals will be properly identified, placed in disposable properly labeled yellow plastic bags, and maintained at refrigeration temperature until disposal in accordance with the institutional and government regulations. Dead animals will be given to the technician for proper disposal. Should the animals be alive after the experiment, they would be put up for adoption by the researchers. H. Is there a non-animal model applicable for the procedure/study? If so, please provide the reasons for not using it. None.
I.
Indicate the names and qualifications of all personnel who will be responsible for conducting the procedures. Name: Diana Patricia D. Bala Student Number: 2014 - 062605 Email Address:
[email protected] Cellphone number: 09175627781 Name: Allyssa Jiselle M. Cabalonga Student Number: 2014 - 062663 Email Address:
[email protected] Cellphone number: 09178808647 Name: Caitlene Joy C. Mejia Student Number: 2014-065613 Email Address:
[email protected] Cellphone number: 09179701913
J. References
K. DECLARATION BY THE RESPONSIBLE PERSON: I ACCEPT RESPONSIBILITY FOR ASSURING THAT THE PROCEDURES/ STUDY WILL BE CONDUCTED IN ACCORDANCE WITH THE APPROVED PROTOCOL. I ASSURE THAT ALL PERSONNEL WHO USE THIS PROTOCOL AND WORK WITH ANIMALS, HAVE RECEIVED APPROPRIATE TRAINING/ INSTRUCTIONS IN PROCEDURAL AND HANDLING TECHNIQUES, AND ON ANIMAL WELFARE CONSIDERATIONS.
RCN AS - UST I ACUC Ani mal Care and Use Statement Pr otocol R eview Form
UST: S037-00-FO01
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UST IACUC Animal Care and Use Statement Protocol Review Form
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I AGREE TO OBTAIN WRITTEN APPROVAL FROM THE INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE PRIOR TO MAKING ANY CHANGES AFFECTING MY PROTOCOL. I ALSO AGREE TO PROMPTLY NOTIFY THE IACUC IN WRITING OF ANY EMERGENT PROBLEMS THAT MAY ARISE IN THE COURSE OF THIS STUDY, INCLUDING THE OCCURRENCE OF ADVERSE SIDE EFFECT.
Signature over Printed Name of the Responsible Person: Date:______________________ Diana Patricia D. Bala
Date:______________________ Allyssa Jiselle M. Cabalonga
Date:______________________ Caitlene Joy C. Mejia
Date:______________________ Asst. Prof. Jose Francis V. Abrantes, M.Sc .,
Date:______________________ Prof. Gina R. Dedeles, Ph.D.
Prof. John Donnie A. Ramos, PhD Dean, UST College of Science
Date:______________________
Daria L. Manalo, DVM Institutional Veterinarian
Date:______________________
RCN AS - UST I ACUC Ani mal Care and Use Statement Pr otocol R eview Form
UST: S037-00-FO01
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