Skip Navigation
Johns Hopkins Bloomberg School of Public HealthCAAT
 

7th Annual 3Rs Symposium: Practical Solutions and Success Stories

rat image © Cathy Schuppli

7th Annual 3Rs Symposium: Practical Solutions and Success Stories

June 4-5, 2020
ONLINE
Register

Downloadable Schedule (PDF)

The 7th Annual 3Rs symposium, co-hosted by the USDA Animal Welfare Information Center (AWIC), NIH Office of Laboratory Animal Welfare (OLAW), the Johns Hopkins Department of Molecular and Comparative Pathobiology, and the Johns Hopkins Center for Alternatives to Animal Testing (CAAT), will be held online on June 4-5, 2020. The goal of this year’s symposium is to bring together experts in replacement, reduction, and refinement of animal experimentation to exchange information with scientists, IACUC members, veterinarians, and animal care technicians about practical solutions and recent success stories to reduce the use of animals in research and improve their welfare.

The format includes 2 days of lectures and Q&A sessions. These lectures give participants a strong foundation in the relevant research underlying breakthroughs in the 3Rs, while Q&A sessions allow participants to receive feedback specific to their own facilities from experts. This symposium has been approved for 9 hours of RACE credits.

AGENDA

Day 1 - June 4th, 2020

8:30 - 8:45
Organizers’ Welcome – Kristina Adams, Patricia Brown, Kathrin Herrmann, Eric Hutchinson

Moderator – Kathrin Herrmann, Center for Alternatives to Animal Testing, Johns Hopkins University


Keynote Presentation

8:45 - 9:45
Beyond Zoonoses: What COVID-19 Can Teach Us About Non-Communicable Diseases and the Importance of Veterinary and Environmental Science for  Human Medicine
Abstract
Barbara Natterson-Horowitz, University of California Los Angeles and Harvard University
Bio (PDF)


Recent Success Stories

9:45 - 10:15
Minibrain Organoids
Abstract
Helena Hogberg, Center for Alternatives to Animal Testing, Johns Hopkins

Helena Hogberg is the Deputy Director of the Center for Alternatives to Animal Testing (CAAT) at the Johns Hopkins Bloomberg School of Public Health, Baltimore, MD. She received her PhD in Toxicology from the Physiology Department at Stockholm University, Sweden in 2009. The scientific work during her PhD was performed at the European Centre for the Validation of Alternative Methods (ECVAM), European Commission, Ispra, Italy and aimed to develop new in vitro approaches to detect chemicals with developmental neurotoxicity (DNT) potential, with a focus on gene expression and electrical activity recordings.

Together with CAAT Director Thomas Hartung she started the current laboratory at CAAT as a Postdoctoral Fellow in 2010. She was later promoted to a faculty position and was appointed Deputy Director of the center in the beginning of 2016. Her current research activity is still in the field of DNT with the use of emerging tools, such as 3D organotypic cell models, induced pluripotent stem cells, and omics (transcriptomics and metabolomics) approaches.


10:15 - 10:45
Innovative Models in Biomedical Research: A Literature Review
Abstract
Laura Gribaldo, MD, PhD, European Commission Joint Research Centre
Bio


10:45 -11:00
Panel Q&A


11:00 - 11:30
Grimace Scale Panel
Abstract
Dale Langford (Mice), Univ. of Washington

Dale Langford is a Research Assistant Professor in the Division of Pain Medicine at the University of Washington in Seattle. She earned her PhD in Experimental Psychology/Behavioral Neuroscience at McGill University (Montreal, Quebec, Canada) under the supervision of Dr. Jeffrey Mogil, where her dissertation focused on social modulation of pain and facial expressions of pain in laboratory mice. Dale conducted her postdoctoral fellowship at University of California, San Francisco where she studied inter-individual variability in the experience of cancer-related pain and co-occurring symptoms. She has continued this work at University of Washington, where she also investigates the impact of provider- and patient-facing interventions for chronic non-cancer pain.

11:30-12:00
Anne Burrows (Nonhuman primates), Duquesne University
Bio (Word Doc)
Abstract

12:00 - 12:15
Panel Q&A



12:15 - 1:00
Lunch Break
 

Hot Topics: Best Data from Design to Publish

Moderator - Cate Pritchard, Office of Laboratory Animal Welfare, National Institutes of Health

1:00 - 2:00
Improving the Design and Reporting of Animal Experiments: the Experimental Design Assistant and the ARRIVE Guidelines 2019 (including Q&A) 
Abstract
Nathalie Percie du Sert, National Centre for Replacement, Refinement and Reduction of Animals in Research (NC3Rs)

Nathalie Percie du Sert is Head of Experimental Design and Reporting at the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), which she joined in 2010. Her programme of work includes the development of the Experimental Design Assistant, an online tool to guide researchers through the design of in vivo experiments and dissemination of the ARRIVE guidelines to improve the design and reporting of animal research.

She holds a PhD from St George's University of London and worked as a post-doctoral researcher in the field of nausea and emesis at the University of California, San Francisco and at the Chinese University of Hong Kong, where she developed expertise in in vivo research and systematic reviews and meta-analysis of animal models.


2:00 - 2:30
PREPARE Guidelines 
Abstract
Adrian Smith, NORECOPA
Bio (MS Word doc) 


2:30 - 3:00
NINDS/NIH Approach to Rigor and Transparency 
Abstract
Shai Silberberg, National Institute of Neurological Disorders and Stroke, NIH

Shai Silberberg is the Director of Research Quality at the NIH National Institute of Neurological Disorders and Stroke (NINDS) leading the Institute efforts to increase the excellence of science and the completeness of research reporting. In addition, Dr. Silberberg is a Program Director at NINDS overseeing basic research related to ion channels and transporters. Prior to joining NINDS, Dr. Silberberg was an Associate Professor at Ben-Gurion University of the Negev in Israel, investigating the biophysical functions and physiological roles of various ion channels.

3:00 - 3:30
Panel Q&A


3:30
Adjourn for Day 1


Day 2: June 5th, 2020

Hot Topics: Best Data from Design to Publish (continued)

Moderator – Kristina Adams, National Agricultural Library, United States Department of Agriculture

8:30 - 9:30
Refinement WIKI and International Culture of Care Network (including Q&A)
Abstract
Adrian Smith, Norecopa
Bio (MS Word doc) 


9:30 - 10:00
The Open Science Landscape at USDA and How the National Agricultural Library Can Help
Abstract
Cynthia (Cyndy) Parr, National Agricultural Library, United States Department of Agriculture

Dr. Cynthia Parr is the acting Assistant Chief Data Officer for the USDA Research, Education, and Economics Mission Area. In this role she is establishing the mission area’s first data governance framework and data strategy in alignment with the USDA enterprise data strategy. At the National Agricultural Library, Dr. Parr focuses on strategic planning and policy for open data, public access, and big data initiatives. She has managed a portfolio of services and platforms including the Ag Data Commons, USDA’s research data catalog and repository, and data management plan policy, guidance, and review services. She continues to build NAL, ARS, and REE advanced data analytics capacity. Dr. Parr received degrees in biology from Cornell University and University of Michigan. She has authored over 60 scholarly articles and 30 software resources and datasets on topics ranging from data standards and semantics to visualization and collaboration systems to evolutionary ecology. She co-chairs the Research Data Alliance Interest Group on Agricultural Data.


10:00 - 10:30
The PRIMatE Data Exchange: an Open Science Resource for Non-human Primate Imaging
Michael Milham, Center for the Developing Brain, Child Mind Institute
Bio (PDF)


10:30 - 10:45
Panel Q&A


Practical Solutions

10:45 - 11:15
How Dogs are Helping Us Understand Cancer 
Abstract
Amy LeBlanc, Comparative Oncology Program, National Cancer Institute
Bio (Word Doc) 


11:15 - 11:45
Image Guided Animal Therapy
Dara Kraitchman, Johns Hopkins Center for Image Guided Animal Therapy

Dr. Dara Kraitchman is a Professor of Radiology and Molecular and Comparative Pathobiology at Johns Hopkins University School of Medicine. She received her VMD in 1992 and her PhD in Bioengineering in 1996 from the University of Pennsylvania. She founded the Center for Image-Guided Animal Therapy (CIGAT) at Johns Hopkins University, which is the first veterinary hospital within an academic medical center without a school of veterinary medicine, to perform advanced diagnostic imaging and minimally invasive procedures in pets on a referral basis. She has spent extensive time understanding the ethical considerations of performing veterinary clinical trials in interventional radiology and cardiology and has lectured on the subject for numerous societies including the Public Responsibility in Medicine and Research (PRIM&R), the International Society for Magnetic Resonance Research (ISMRM), Society for Cardiovascular Magnetic Resonance (SCMR), and American Heart Association (AHA). With over 100 peer reviewed publications in translational research in imaging, cardiology, and oncology, she is the first veterinary recognized as a Fellow of the American College of Cardiology (FACC) and ISMRM.


11:45 - 12:00
Panel Q&A


12:00 - 12:45
Lunch Break


Moderator – Eric Hutchison, Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine

12:45 - 1:15
Behavioral Training as Part of the Health Care Program 
Abstract
Steven Schapiro, Department of Comparative Medicine, MD Anderson Cancer Center

Dr. Schapiro is associate professor of comparative medicine and chief of the Section of Primate Behavior and Environmental Enrichment in the Department of Comparative Medicine of The University of Texas MD Anderson Cancer. For more than 30 years, his research group has been conducting studies focused on the behavior and welfare of captive rhesus monkeys and chimpanzees, including collaborative research projects funded by NIH and NSF. An important emphasis of his research program is to provide non-human primates with opportunities to voluntarily participate in their own care. He is a co-founder of the Primate Training and Enrichment Workshop and the founder of the Primate Behavioral Management Conference. He is the editor of the Handbook of Primate Behavioral Management and co-editor of the Handbook of Laboratory Animal Science. He is a past-president and former treasurer of the American Society of Primatologists, and the former treasurer and vice president for membership of the International Primatological Society. He currently organizes the ‘3Rs for the CPRC’ (a/k/a Project Monkey Island), an effort to revitalize Cayo Santiago after the devastating effects of Hurricane Maria.


1:15 - 1:45
Pig Handling 
Abstract
Derek Brocksmith, Sinclair Biosciences

Derek has spent the last several years developing the miniature swine program and working with sponsors on the selection of animal models and conduct of nonclinical research studies. Focusing on behavior, Derek works with clients to minimize human and animal stress developing protocols that are sustainable for working with swine. Mr. Brocksmith worked at Sinclair Research Center as an animal and veterinary technician prior to serving at Sinclair BioResources as General Manager.


1:45 - 2:15

Hydrophobic Sand as an Alternative to Metabolic Cages 
Abstract
Jessica Hoffman, Uniformed Services University of the Health Sciences

Jessica Hoffman’s greatest passion is in integrative, translational research in the field of mental health, with a strong interest in policy, ethics, and advocacy for better research and treatment. She received a BS in Biochemistry and Molecular Biology and a BA in Psychology from UMBC in 2008, and her PhD in Neuroscience from UMB-SOM in 2014, where her thesis work used cellular and behavioral techniques to discover a sensitive period in which inflammation affected cerebellar development and led to an increased risk of autism-like behavior in the rat. Her postdoctoral work with the National Institutes of Health (NIH, NIMH and NIAAA used cell lines from human clinical trials to integrate physiological and biochemical/genetic research to achieve a more complete understanding of hormone-related mood disorders such as perimenopausal depression (PMD) and premenstrual dysphoric disorder (PMDD). As a Research Biochemist at AFRRI she uses a rat model of embedded metals (i.e. shrapnel wounds) to examine the health effects of military-relevant metals across a multitude of tissues, collaborating with the Maryland VA program and the FDA to identify health risks and early warning biomarkers to improve treatment policy. In addition to her research, she enjoys serving on the AFRRI/USU IACUC.

2:15 - 2:45
Panel Q&A

Moderator – Patricia Brown, Office of Laboratory Animal Welfare, National Institutes of Health


2:45 - 3:15
Mouse handling limits the impact of stress on metabolic endpoints 
Abstract
Parinaz Mahbod, University of Cinncinnati
Bio (PDF)


3:15 - 3:45
State of the Art Rodent Handling: Can a Little Time Go a Long Way for Welfare? 
Abstract
Brianna N. Gaskill, Purdue University

Brianna received her BS from Kansas State University in 2004 and her PhD in animal behavior and well-being from Purdue University in 2011. After graduation, she spent 2.5 years as a postdoctoral research scientist at Charles River, studying the behavior and well-being of laboratory rodents and is currently an assistant professor of animal welfare at Purdue University. Her previous research has covered behavioral and physiological thermoregulation of mice in laboratories and its impact on mouse well-being. Additionally she has been involved in developing new and improved types of cognitive testing for mice that are used in psychiatric and neuroscience research. Brianna’s other research interests include the development of refined methods in behavioral research, rodent well-being, and the scientific impact of well-being problems in laboratory animals. Brianna has published in the behavior and well-being, laboratory animal, and experimental psychology literatures and has given presentations on her work nationally and internationally. Her research contributions have been acknowledged by receiving the highly commended paper prize from the NC3R’s in 2015, the prize for exceptional service in laboratory animal science from the Swiss Laboratory Animal Science Association, and the New Investigator Award from the International Society for Applied Ethology in 2013. Recently she was honored to consult with NASA about mouse welfare on the international space station.

Personal Web Page: Google Scholar PageMouse Ethogram



3:45 - 4:00 
Panel Q&A
 

4:00
Adjourn Day 2
 


ABSTRACTS

From Zoonoses to Zoobiquity: An Expanded View of the Connection Between Human and Animal Health
Barbara Natterson-Horowitz

The emergence of non-communicable diseases (NCDs) as the leading cause of death in humans heightens the urgency of increased physician engagement in One Health and Planetary Health initiatives. The significant influence of shared environments on the occurrence of NCDs in human and non-human animals points to a critical need to expand physician awareness of One Health and Planetary Health efforts. Using examples from cardiovascular medicine, cancer and psychiatry, the talk presents several high-impact human NCDs in a One Health context and examples of educational, research and clinical initiatives developed to increase the human medical profession in One Health and Planetary Health communities.  


Minibrain Organoids
Helena Hogberg

Lately it has become evident that more complex in vitro cell models, such as three dimensional (3D) test systems are essential to reproduce the architecture and function of an organ. Many reports indicate that the third dimension in cell cultures is important for improvement of drug discovery, disease modelling and toxicity testing, as these models are much closer to in vivo compared to two-dimensional cultures. This holds especially true for the central nervous system (CNS) which consist of numerous different cell types and the cell-cell interactions are of key importance for brain development and function. In addition, more than 94% of CNS drugs fail in clinical trials why new more human relevant approaches are needed.

Our group was funded by the NIH to develop a 3D brain model from human induced pluripotent stem (iPSCs). The model consists of several different cell types, such as neurons, astrocytes, and oligodendrocytes that reproduce neuronal-glial interactions and connectivity. In addition, it has shown critical elements of neuronal function—synaptogenesis and neuron-to-neuron (e.g., spontaneous electric field potentials) and neuronal-glial function (e.g., myelination). The model has been used for numerous different applications, including (developmental) neurotoxicity, Parkinson’s disease, cancer, resilience, blood-brain barrier, autism, Down syndrome, inflammation, Zika, and other viral infections. The possibility of infecting this human iPSCs model with viruses such as Zika and JC-virus is especially exciting, as very few animal and in vitro cell models been able to recapitulate these pathologies. The model has the potential to become a human-relevant alternative to animals in toxicology and disease.


Innovative Models in Biomedical Research: A Literature Review
Laura Gribaldo

Mammalian models have served as a basis for biomedical research, safety testing and development of drugs over the past decades. Nevertheless, these models may yield results that cannot always be translated into the human in vivo situation. The biology and physiology of rats and mice, the animal species mostly used, may differ significantly from that of humans. Furthermore, currently around 90% of drugs fail in clinical trials, mainly due to lack of efficacy or toxicity in humans that was not predicted by animal tests. Therefore, scientific concerns about the predictive power of animal models are driving forces for the development of animal-free, human relevant approaches in different branches of life sciences. Great advances have been recently made in life science technologies and computer science. Today scientists and researchers can count on a toolbox of a variety of non-animal approaches comprising in vitro techniques (organ-on-a-chip, microphysiological systems, three-dimensional organoids, robot-assisted high-throughput systems, high-content imaging technologies, 3D bioprinting, and DNA or RNA microarray screening) and computational tools, such as machine learning and artificial intelligence. Tools and models that are more relevant to human biology will increase our understanding of human diseases and the search for effective therapeutics. A first effort to gather existing knowledge about non-animal models of highly prevalent human diseases has been made by the Joint Research Centre of the European Commission. The final goal is to disseminate and improve knowledge sharing on potentials and limitations of human based models at different levels: scientific communities, universities and secondary schools, national committees for animal welfare and the public at large.


Grimace Scale Panel
Dale Langford

Assessment of pain in mice and other animals is complex, yet vital for biomedical research and veterinary care. Pain assessment in laboratory animals traditionally relies on evoked responses, which may not adequately capture the most prevalent and bothersome aspect of chronic pain (i.e., spontaneous, non-evoked pain). In fact, the lack of effective clinical treatments for pain may - at least in part - be due to the inadequate predictive value of animal models. Ten years ago, we developed the Mouse Grimace Scale (MGS) to evaluate facial expressions of pain in laboratory mice. Since that time, grimace scales have been developed for rats and several other mammalian species. This presentation will cover the development of the MGS, its psychometric properties, factors that may modulate grimacing, advantages and disadvantages of grimace scales, and their impact on pain measurement.

Anne Burrows

Facial expressions are the primary mechanisms that primates use to visually communicate with one another and these facial movements are produced by mimetic musculature. Rhesus macaques (Macaca mulatta) are one of the most commonly studied primates, in laboratory and field settings, and are used for biomedical research and modeling the evolution of human social behavior. Because rhesus macaques typically live in large, multi-tiered social groups, facial expressions are a prominent feature of their social interactions. A better understanding of the gross and microanatomical aspects of their mimetic musculature can inform our conceptualization of how they use facial expression. To these ends, the current study describes the gross anatomy of rhesus macaque mimetic musculature, the microanatomical arrangements of these muscles, their physiology, and the major facial displays used by rhesus macaques.


Improving the Design and Reporting of Animal Experiments: the Experimental Design Assistant and the ARRIVE Guidelines 2019
Nathalie Percie du Sert

Many factors influence the reproducibility of preclinical experiments, with issues around experimental design and reporting estimated to account for half of irreproducible research. Researchers often have limited knowledge and understanding of experimental design and statistics, and may not appreciate the importance of rigorous methodology. This has an impact on the way we design and conduct experiments and it also affects the way we assess manuscripts and grant proposals in our capacity as journal editors, reviewers and funding panel members. The NC3Rs has developed a suite of tools to support researchers. This includes the recently revised ARRIVE guidelines and accompanying Explanation and Elaboration document, and the online Experimental Design Assistant which provides bespoke feedback on individual experimental plans. These resources provide extensive guidance on how to design, conduct and analyse animal experiments, and what crucial information to report in scientific publications. This presentation will include a live demonstration of the Experimental Design Assistant.


PREPARE Guidelines
Adrian Smith

Efforts to improve the scientific rigour and reproducibility of animal experiments have tended to focus on improved reporting and "experimental design"—where the focus is often on the more mathematical elements of design such as randomization, blinding and statistical analysis.

The PREPARE guidelines were written by Norwegian and British scientists as the result of dialogue over a 30-year period, from courses in Laboratory Animal Science for researchers and technicians, supervision of animal research and through management of laboratory animal facilities, including AAALAC accreditation. The name PREPARE implies that we encourage scientists to seek collaboration from day 1 of planning with the animal facility where they intend to work. This ensures that all aspects of the planned experiment, including the practicalities, timeframe, distribution of labour and costs, are discussed with all those who will be involved.

PREPARE consists of 15 main topics, organised in the form of a checklist, which has been translated into 20 languages. The checklist is not meant to add to the administrative burden, and should be used as a basis for dialogue rather than as a mandatory requirement.

Importantly, the checklist is supported by a website where each of the 15 topics is described in more detail. The website is updated continuously with links to new resources within each field. We have made a 3-minute cartoon film illustrating the main principles in PREPARE, making comparisons with the aviation industry with their excellent record of safety and reliability. PREPARE is available at https://norecopa.no/PREPARE

Reference

Smith AJ, Clutton RE, Lilley E, Hansen KEA & Brattelid T (2017): PREPARE: Guidelines for Planning Animal Research and Testing. Laboratory Animals, 52(2): 135-141. https://journals.sagepub.com/doi/10.1177/0023677217724823


NINDS/NIH Approach to Rigor and Transparency
Shai Silberberg

The quality of research depends on the rigor with which researchers conduct studies and control for potential biases. Numerous publications and meetings have focused on the observation that published research results are often not reproducible. Although there are many possible causes for this issue, inadequate experimental design, insufficient reporting of research methodologies, and publication bias appear to be important contributing factors. Evidence for this deduction will be presented, as well as actions taken by the US National Institutes of Health (NIH) and by publishers to address the issues.


Refinement WIKI and International Culture of Care Network 
Adrian Smith

This presentation will highlight two recent collaborative projects designed to refine animal studies and to improve the atmosphere in which we work—for the benefit of both the animals and personnel.

All those who have participated in congresses or email discussion forums know that there are many people developing or using refinements to animal care and use which have not been published in the scientific literature. This is often because of financial or time constraints, or because the refinement was not considered worthy of a publication in its own right.

To bridge the gap between scientific papers and discussion groups, we have constructed a Refinement Wiki (https://wiki.norecopa.no/), where improvements can be posted. We hope that it will also function as a hub where those investigating the effects of a potential refinement can identify collaborators, and that it will encourage colleagues to share experiences or develop new strategies to solve a current problem.

The contents of the Wiki are not curated. The quality of the Wiki is determined by registered bona fidemembers of the research animal community. No one else can add, delete or comment upon material. Wiki content is retrievable from Norecopa's main search engine, and the Wiki has also its own search engine. Those interested in participating are encouraged to take contact.

There is increasing focus internationally on fostering a climate of care around animal research - also known as a "culture of care". In Europe this is mentioned specifically in the text accompanying the EU Directive on the protection of animals used for scientific purposes. Culture of care means a commitment to improving animal welfare, scientific quality, care of the staff and transparency for the stakeholders. The EU Commission has produced guidance documents on how to implement such a culture, and these have been endorsed by all Member States.

An international Culture of Care Network was established following the FELASA congress in Brussels in 2016. Norecopa acts as the host website for this network: https://norecopa.no/CoC. Members of the network include animal technicians, veterinarians, members of animal welfare bodies, representatives of competent authorities, communications experts and members of animal welfare organisations. The primary aim of the network is to share and publish examples of activities fostering a culture of care. Emphasis is also placed on methods for improving work conditions for all those who care for or use animals in research and testing. The website is used to post practical examples of how to implement a culture of care.

Closely related to a culture of care is the concept of a Culture of Challenge: looking for the acceptable, rather than choosing the accepted. This is much easier to achieve if a caring atmosphere has already been created at the workplace.



The Open Science Landscape at USDA and How the National Agricultural Library Can Help
Cynthia (Cyndy) Parr

Both academia and governments increasingly promote open science as a way to accelerate progress and innovation. In this talk, we will present USDA public access policies and open data plans that impact research publication and data sharing. We introduce some of the National Agricultural Library (NAL) services that support these initiatives and consider the implications and opportunities for minimizing the use of animals in research. Specifically we will review the recent activities of USDA’s extramural and intramural research agencies in response to the 2013 Holdren memo, requiring public access to the products of USDA-funded research. The National Agricultural Library provides PubAg (pubag.nal.usda.gov) and AgDataCommons (data.nal.usda.gov) as central access points for full text and data. In addition, NAL provides guidance and consultation on data management planning, and coordinates with land grant university libraries. Increased access to full text and data and published methodologies should benefit researchers in several ways: by increasing citations, by improving the ability to discover and re-use relevant protocols such as animal use alternatives, and by improving the ability to conduct systematic reviews and meta-analyses.


How Dogs are Helping Us Understand Cancer
Amy LeBlanc

Comparative oncology clinical trials play an important and growing role in cancer research and drug development efforts. These trials, conducted in companion (pet) animals, allow assessment of novel anticancer agents in a veterinary clinical setting that supports serial biologic sample collections and exploration of dose, schedule, and corresponding pharmacokinetic/pharmacodynamic relationships. Further, an intact immune system and natural co-evolution of tumor and microenvironment support exploration of novel immunotherapeutic strategies in these veterinary patients. Significant improvements in our collective understanding of the molecular landscape of veterinary cancers, mainly in dogs, have occurred in the last 10 years, facilitating the translational research process and supporting the inclusion of comparative studies into the drug development paradigm. Further, recent clinical trials carried out in pet dogs demonstrate how this approach can assess efficacy in a variety of settings, including but not limited to single agent or combination response rates, inhibition of metastatic progression, and randomized comparison of multiple agents in a simultaneous head-to-head fashion. Such comparative oncology studies have been purposefully included in the developmental plan for several FDA-approved and up-and-coming anticancer drugs. Challenges for this field include keeping pace with technology and data dissemination/harmonization, improving annotation of the canine genome and immune system, and generation of canine-specific validated reagents to support integration of correlative biology within clinical trial efforts.

Burton JH, Mazcko C, LeBlanc A, et al. NCI Comparative Oncology Program Testing of Non-Camptothecin Indenoisoquinoline Topoisomerase I Inhibitors in Naturally Occurring Canine Lymphoma. Clinical Cancer Research; Published OnlineFirst July 30, 2018; DOI: 10.1158/1078-0432.CCR-18-1498.

Naik SN, Galyon GD, Jenks NJ, … LeBlanc AK. Comparative oncology evaluation of intravenous recombinant oncolytic Vesicular Stomatitis Virus therapy in spontaneous canine cancer. Molecular Cancer Therapeutics 2018 Jan; 17 (1): 316-326.

Saba C, Paoloni M, Mazcko C, … LeBlanc AK. A comparative oncology study of iniparib defines its pharmacokinetic profile and biological activity in a naturally-occurring canine cancer model. PloS One 2016 Feb 11;11(2):e0149194. doi: 10.1371/journal.pone.0149194.

LeBlanc AK, Mazcko C, Khanna C. Defining the value of a comparative approach to cancer drug development. Clinical Cancer Research. Published OnlineFirst Dec 28, 2015; doi: 10.1158/1078-0432.CCR-15-2347.

LeBlanc AK, Breen M, Choyke P, … Khanna C. Filling in the Gaps for Man’s Best Friend: Perspectives from a National Academies Institute of Medicine Workshop on Comparative Oncology. Science Translational Medicine 2016; 8 (325) [epub ahead of print]


Behavioral Training as Part of the Health Care Program
Steven Schapiro

Reinforcement Training (PRT) techniques are employed to provide research animals with opportunities to voluntarily participate in a variety of husbandry, veterinary, and research procedures. PRT allows the animals to control their environments and to make meaningful choices. PRT has proven particularly effective for the health care and behavioral management of nonhuman primates living in research settings. This talk will include a very brief discussion of the Stimulus-Response-Reinforcement contingency that is the basis for PRT, followed by a number of examples of ways in which PRT can be used to facilitate the health care of nonhuman primates. By providing the animals with control over the way they are treated, we can provide them with a more functionally appropriate captive environment, enhance the quality of care that they receive, and even, implement treatment plans that would not be possible if the animals were not trained.


Pig Handling
Derek Brocksmith

We use targeting to overshadow fear responses in Miniature Swine (MS). Targeting is part of the unique Sinclair’s proprietary behavior program called Behavioral Enhancement System™(BES). BES is an integral approach to MS behavior instituted and developed by Sinclair to provide well-conditioned and easily trained MS to clients. We use targeting to replace an unacceptable behavior such as a fear response with an acceptable one that involves a touch in response to the same stimulus. The handler first finds highly prized reinforcers, often a food reward, and gradually teaches the MS that is afraid to interact with people to want to interact with people. In this case the MS is taught to "touch" a target-stick for food-reward. It is walked a short way following the target stick and asked to "touch" the target, then food-reward. Once the MS can predict a reward is coming when interacting with people, we can condition a MS to cooperate for SOPs such as loading a cart. Every week, technicians participate in a POC (Performance On-Cue) tournament with a MS they have trained. MS follows targeting cues to navigate through a maze of obstacles. Fears and anxiety are eliminated because the MS is taught another behavior (touch) that is more enjoyable or pleasant to exhibit in the presence of the stimulus (human interactions, obstacles, and cart) that elicits the abnormal behavior, i.e. a fear response.


Hydrophobic Sand as an Alternative to Metabolic Cages
Jessica Hoffman

A common method for rodent urine collection requires a metabolic cage, exposing animals to extended periods of isolation in an unfamiliar cage with a wire-mesh floor. A new method involving hydrophobic sand, a material similar to bedding, has become available but has not been extensively compared with metabolic cages in regard to collection efficiency or stress. Using a within-subjects crossover design, we examined differences in stress markers, urinary markers, and urine volume of healthy male Sprague–Dawley rats during multiple collection sessions in hydrophobic sand and metabolic cages. Stress response markers of weight loss, fecal pellet output, or corticosterone did not differ between hydrophobic sand and metabolic cages, and observed behavior suggested that sand may be less stressful than metabolic cages. All clinically relevant urinary markers examined were normal, with no differences between collection methods. Total urine volume collected was greater from the metabolic cage than sand in most sessions, but the volume collected during the shortest session accounted for 62% of the total volume collected during the longest session. We found no contamination of the sand with several common metals of interest. We also isolated extracellular vesicles (EVs) containing miRNAs from the urine and found no significant differences in particle size, particle concentration, total RNA, or the type and abundance of miRNAs due to urine collection method. Our results suggest that hydrophobic sand is a refinement of urine collection methods for rats that decreases isolation time, risk of injury, and stress and maintains the integrity of urine samples.

Hoffman, J.F., Vechetti Jr., I.J., Alimov, A.P., Kalinich, J.F., McCarthy, J.J., Peterson, C.A. (2019) Hydrophobic sand is a viable method of urine collection from the rat for extracellular vesicle biomarker analysis. Molecular Genetics and Metabolism Reports. (e-pub, vol 21, article 100505, DOI 10.1016/j.ymgmr.2019.100505)

Hoffman, J.F., Fan, A.X., Neuendorf, E.H., Vergara, V.B., & Kalinich, J.F. (2018) Hydrophobic sand versus metabolic cages: a comparison of urine collection methods for the rat (Rattus norvegicus). Journal of the American Association for Laboratory Animal Science. 57(1):1-7

Hoffman, J.F., Vergara, V.B., Mog, S.R., & Kalinich, J.F. (2017) Hydrophobic sand is a non-toxic method of urine collection, appropriate for urinary metal analysis in the rat. Toxics 5(4), 25

Disclaimer: The views expressed in this presentation are my own and do not reflect the official policy or position of the Armed Forces Radiobiology Research Institute, Uniformed Services University for Health Sciences, Department of Defense, or US Government. I have no conflicts of interest to report. The use of specific products in this presentation does not constitute an endorsement by the US Government.


Mouse handling limits the impact of stress on metabolic endpoints
Parinaz Mahbod

Reproducibility is a factor pivotal to determining the validity of any study. For animal studies, reproducibility is increasingly appreciated to be enhanced by minimizing the confounding effects of variable baseline stress. Some factors particularly important for animals include noise, odor, light/dark cycle, housing, and handling. As a focus of our laboratory is studying the impact of stress on both endocrine, specific neuroendocrine pathways and behavioral endpoints, we aimed to establish experimental conditions in which the confounding effects of stress would be reduced so that the effects of specific interventions could be more reproducibility be tested. 

Based primarily on published observation from the work of Hurst and West (Nature Methods 2010), we designed a series of studies to compare physiology and behavior of mice comparing their “cup” handling vs. standard “tail-picked” method. As their studies had primarily behavior outcomes, we sought to include measures of glucose homeostasis and stress hormones. First, tail whole blood glucose and plasma corticosterone were measured after elevated plus maze (EPM) test vs. control. Cup-handled animals had lower basal and stimulated plasma corticosterone levels than the unhandled animals. In addition, average fasted basal blood glucose was ~100 mg/dl and 160 mg/dl respectively. Having established conditions in which baseline stress could better controlled for, we elected to study the effect of Desacyl Ghrelin, a hormone known to exert an anxiolytic effect. Interestingly, we found an anxiolytic effect of Desacyl Ghrelin only in handled animals. This result provides an example of how without providing a baseline state that is low in stress, the impact of a specific intervention is not detected.  

In summary, our results reinforce the importance of establishing conditions in which the effects of baseline stress are accounted for in any experimental design. Specifically, as initially recommended by Hurst and West, we advocate minimizing the impact of confounding background stress by prior habituating experimental mice by cup handling. 


State of the Art Rodent Handling: Can a Little Time Go a Long Way for Welfare?
Brianna N. Gaskill

Handling laboratory rodents and other common husbandry and experimental procedures can be a source of stress leading to the development of aversive responses to handling or human presence that are suggestive of emotional distress. Aversive handling of animals can lead to a fearful relationship with humans, making handling difficult, and increasing the risk of injury for both handler and animal, thus affecting human and animal welfare. Recent research has provided evidence that new handling techniques such as tickling, tube handling or, gentling with associative conditioning are effective at reducing fear of humans when rodents are handled for common husbandry activities and medical and research procedures such as injection. Some techniques such as tickling and associative conditioning techniques are even perceived as positive by rats, which improves the human-rodent relationship and rat welfare. The goal of the seminar is to review new state of the art rodent handling techniques and provide scientific data to support their effectiveness in improving rodent welfare as well as data quality.