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Both the Word and PDF versions contain useful internal and external hyperlinks. Guidelines for the use of fishes in research. American Fisheries Society, Bethesda, Maryland. Their policies and standpoints are based primarily on information developed through scientific practices, but they also reflect ethical concerns, including the conservation of the diversity and abundance of fish populations, and respect for life and life processes.
Further, these societies believe that their members are responsible not only for advancing scientific knowledge and understanding of fish and fisheries but also for improving human appreciation for these animals and the industries that they support. All three societies actively promote research and the dissemination of information derived from that research.
They also advocate respect for life processes, respect for the forms of life within various ecosystems, and the humane treatment of animals used in research investigations. Fishes are worthy of experimental and observational research: Further, fishes are economically important through recreational and commercial activities in that they provide an important source of food for humans and other animals and are popular to catch and to observe.
The authors of the Guidelines for the Use of Fishes in Research referred to hereinafter as the Guidelines are scientists, have respect for life, and are professionally trained in a multitude of disciplines. The current version is in answer to the call for a periodic document review and update, with a similar suggestion noted herein. The understanding and welfare of animals used in research can be served best by using a multidisciplinary approach in which data and expertise are derived from such disciplines as ecology, behavioral studies, nutrition, genetics, toxicology, chemistry, endocrinology, physiology, anatomy, and fish health.
At the same time, understanding that research is conducted in a variety of human cultural settings is important. Ideally, scientific procedures, analytical methods, data interpretations, and conclusions based on scientific studies should be consistent across all cultures; however, personal belief systems can and do influence concepts regarding which practices and methods are, or are not, consistent with humane treatment of animals.
The and Guidelines not only reflect the scientific expertise of both UFR Committees but also provide a framework for the promotion of scientifically valid research on fish and fish habitats and for research that is conducted in a manner acceptable to the social communities within which the research takes place. The Guidelines address both field and laboratory research with fishes and will serve as a resource document on topical themes. Specific information in response to United States laws is a focus here, yet these Guidelines can be applied and adapted internationally by investigators working within their own institutional infrastructure with regard to animal care and use committees.
Internet pathway links to various Web sites and documents are included; however, such pathways to online media may change. The UFR Committees that produced the and Guidelines gave generously of their time and expertise by developing, updating, and revising sections.
We thank peer reviewers: Finally, we acknowledge Bill Fisher, past AFS President, for appointing the current UFR Committee and current President Robert Hughes in helping AFS continue its tradition of advancing sound science and disseminating science-based fisheries information for the global protection, conservation, and sustainability of fishery resources and aquatic ecosystems.
Hughes are also thanked for their careful reviews. Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the U. The and Guidelines were developed to provide a structure that advances appropriate attention toward valid experimental designs and procedures with aquatic animals while ensuring humane treatment of the experimental subjects. At a practical level, the Guidelines are intended to provide general recommendations on field and laboratory endeavors, such as sampling, holding, and handling fishes; to offer information on administrative matters, including regulations and permits; and to address typical ethical concerns, such as perceptions of pain or discomfort experienced by experimental subjects.
These Guidelines must be recognized as guidelines. They are not intended to provide detailed instructions but rather to alert investigators to a broad array of topics and concerns to consider prior to initiating study. At a comprehensive level, the principles upon which these Guidelines are based are broadly applicable, and many of the described practices and approaches can be adapted to situations involving other aquatic animal species and conditions.
Understanding the differences between fishes and other vertebrates, especially mammals, is critically important to conducting scientifically sound research with fishes.
Disparities in life histories and mortality rates in fishes versus other vertebrates are critical in designing sustainable sampling levels in fish populations. The UFR Committee points out that 1 compared to mammalian populations, adult populations of many fish species persist despite very high natural mortality rates in juvenile stages by virtue of the fact that most species lay thousands or tens of thousands of eggs; 2 because of these mortality patterns, research on fishes, especially field research or research on early life stages, can involve, and often requires, much larger numbers of research subjects than does research on mammals; and 3 the animal handling and husbandry requirements for fishes are fundamentally different from those for mammals and other vertebrates, in general.
Policies, regulations, and recommendations developed for research on mammals, birds, reptiles, or even amphibians are frequently inappropriate for research with fishes. The Guidelines also address some of the ethical concerns that motivate guidelines used for research with other vertebrates, while being mindful of the unique physiology and general nature of fishes.
The Guidelines were developed for general use by investigators within the United States; therefore, the roles, responsibilities, and informational needs of Institutional Animal Care and Use Committees IACUCs were given specific attention. Investigators from other nations who read this document may disregard specific references to U.
The principles described herein, however, are applicable to research on fishes regardless of geographic location. Investigators in other nations may benefit by modifying any of the specific provisions pertaining to the United States, thereby adopting guidelines consistent with the laws and regulations of their own government.
The UFR Committee urges that the Guidelines be endorsed and adopted adapted, where necessary by those state and federal authorities with regulatory responsibilities for fishes, offices with federal oversight e. Experimental studies using live, intact organisms continue to play an essential role in developing knowledge and better understanding of life processes, life forms, and the environments in which they occur. The enormous evolutionary radiation of fishes comprises at least 27, species Nelson Fishes exist in a multitude of forms and have many unique physiological, behavioral, and ecological specializations.
Fishes occupy a variety of niches in virtually every kind of aquatic habitat. Understanding their biology cannot be accomplished in the absence of experimentation with live, intact animals. Among the reasons for studying fishes are the following: Instead, broad guidelines building on the most current, scientifically valid information are provided in the Guidelines for interpretation and application by various investigators who frequently are the authority on the species or systems involved in their studies.
Ultimate responsibility for the ethical and scientific validity of each study and the methods employed rests with the investigator; however, government agencies, reflecting the beliefs and values of the citizenry and acting on their behalf, often require that investigators follow codes which prescribe acceptable strategies, techniques, facilities, conditions, and post-experimental disposition of animals used in research. Some individuals have argued that fishes should not be included under laws and policies aimed primarily at mammals and birds; however, the Health Research Extension Act HREA of Public Law , http: The ILAR guidelines specifically call for the development of detailed information by knowledgeable groups.
Generally, scientific societies with expertise on particular classes of vertebrates are considered to be the most appropriate sources for the supplemental information needed to implement existing policies. The and Guidelines emphasized field research, and the Guidelines added material on laboratory research with fishes. The Guidelines expands the Guidelines by providing updates with new knowledge and information brought forward over this past decade.
Certain general considerations apply to nearly all research investigations on fishes, whether conducted in the field or in a laboratory setting.
This section introduces concepts and procedures that can be adapted to the situation and circumstances for each investigator.
Research studies should have well-understood and justifiable objectives that address, within the context of the research discipline, basic needs for knowledge and understanding of the world in which we live and the particular resource under consideration. In cases where the biology of fishes proposed for study is well known, a study hypothesis should be articulated so that the use of fishes is focused on addressing that hypothesis.
If the biology of a fish species is not well known, general sampling and other observational uses of fishes i. Research quality relies on the carefully communicated questions that can be addressed by scientific methods and on the development of research procedures that are quality controlled, publishable, and repeatable.
Guidelines have been published for the improved reporting on animal research and for increased reproducibility Kilkenny et al. The validity of research results is affected by the experimental design, the analytical procedures employed, and the quality and health status of the experimental subjects. The quality and appropriateness of the fishes used, both the species and the individuals, can seriously influence results and conclusions, thereby having dramatic effects on the number of animals needed and the number of times that the study is repeated.
These effects, in turn, will have important animal welfare and financial implications. Research scientists have long recognized the importance of animal welfare considerations; however, formal guidelines for the use of fishes in research were not common in the United States prior to , when requirements that research proposals obtain the approval of an IACUC were imposed Public Law , http: Although the principles and procedures described in these Guidelines have been designed to address requirements imposed by IACUCs in the United States, the general concepts are applicable to investigators around the globe.
When approval by an IACUC is required, investigators must prepare written statements or requests for animal use that ensure that all applications, proposals, and actual research will meet certain basic requirements Silverman et al. Safeguards include limiting unneeded replications of research, use of appropriate species and number of animals, an adequate experimental design, and method of euthanasia or disposition of the animals after the completion of the study.
Investigators must be familiar with the species to be studied, or with closely related surrogates, so as to be able to provide environmental conditions essential for the well-being of the experimental subjects and to be able to recognize their responses to disturbances, including capture, restraint, or other changes in environmental conditions that may be applicable to the particular study. Project Quality Assurance Plans PQAPs and standard operating procedures SOPs , while not required by many universities or nongovernmental institutions, are nevertheless recommended as useful tools for maintaining overall research quality.
A PQAP should include a description of the project design and approach to the problem, a statement of anticipated deliverables, and a description of how the data will be reported for instance, as individual measurements or in a reduced form. The PQAP documents the principles, policies, organizational responsibilities, objectives, implementation actions, and accountability procedures that will ensure an appropriate quality process throughout a research project.
Emergency contacts and contingency plans for animal care may be included in a PQAP. The PQAP should identify how unauthorized laboratory personnel or research project areas will be controlled and biosecurity maintained.
For research involving field sampling, the PQAP should include a sampling plan outlining the methods, equipment, and procedures, as well as safety procedures for personnel. For laboratory research, sample preparation and analytical methods should be specified. SOPs for routine, specialized, or unpublished methods should be identified.
Data quality objectives should be identified for each measured parameter, and quality control procedures to be used, whether in the field or under laboratory conditions, should be stated. Corrective actions should be specified should problems arise. Finally, a quality assurance QA report should be generated at the conclusion of the study.
This report should include a summary of the PQAP, results of technical systems and performance evaluation audits, corrective actions taken and results of those actions, data quality assessments precision, accuracy, representativeness, completeness, comparability, and reporting limits , a discussion of whether the QA objectives were met, and limitations on use of data collected during the research.
SOPs document routine or repetitive technical activities, or those that occur occasionally yet need to be followed in the exact fashion as the previous time.
Ideally, SOPs ensure that work is done correctly the first time, thereby reducing unnecessary repetition and costs. If research data are generated in a setting where the same procedures are used yet personnel change with time, SOPs help maintain consistency. Some institutions require SOPs for assuring that job applicants can perform the needed tasks. SOPs also can form an essential part of effective training programs.
General SOPs, not specific to individual studies, may be established as basic procedures for entire research laboratories or institutions.
Examples of specific technical tasks for which SOPs are useful in conducting research with fishes include blood sampling, vaccination protocols, procedures for electrofishing, preparation of solutions for use as sedatives, and techniques for collecting meristic data.
Policies requiring adherence to SOPs and PQAPs in research protocols are to allow for the development of new procedures and for the revision or expansion of established procedures whereby new study techniques are developed. Inherent in the use of animals in research is the responsibility for efficient, effective design of experimental studies, wherein healthy animals without abnormal physiognomies and behaviors will facilitate study results, and for humane treatment of experimental subjects Klontz and Smith ; Snieszko ; Klontz ; NRC Study objectives should be clearly stated, and explanations should be provided on the need for the type and quantity of data to be collected, as well as what will constitute an end to the experiment.
The number of animal subjects required for an investigation will depend on the questions being explored.
Field studies and laboratory studies typically require greatly different statistical designs. The life stage of the fish used in each study will also affect the numbers needed. Studies of early life stages may require very large numbers of individuals. In all cases, studies should be designed to use the fewest animals necessary to reliably answer the questions posed.
The use of adequate numbers to establish variance and to ensure reliability is essential so as to prevent needless repetition of the study ASIH et al. Pseudoreplication can result from wrongly treating multiple samples from one experimental unit as multiple experimental units or from using experimental units that are not statistically independent Heffner et al.
Statistical power analysis can improve designs of experiments Peterman Conducting statistical power analyses ensures the development of study designs that have the appropriate statistical power to accomplish research objectives. In laboratory studies, experimental endpoints, other than death of the experimental subjects, should be developed unless mortality is required by the study protocol.
The use of mortality as an endpoint is appropriate when one or both of the following criteria are met:
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