https://insidescientific.com/webinar/improving-animal-modeling-24/7-home-cage-monitoring-bioexclusion-biocontainment-mouse-housing-system-tecniplast Recently, a surging response to the COVID-19 pandemic has led to an exponential increase in study support for biocontainment and bioexclusion research. Mouse models are being rapidly developed in both areas, and biosafe housing of these animal models is critical. Additionally, non-invasive home cage monitoring can improve the translational value of these research models. Locomotor activity patterns can be monitored 24/7 as a diagnostic tool for biosecurity studies. Researchers, staff and animals alike will also benefit from a decreased need for animal handling, caging manipulations and animal monitoring. This webinar will be most valuable for institutions where biocontainment and bioexclusion work is being considered or conducted, and for researchers who wish to better understand what can be achieved through continuous measurement of animal welfare based of use of non-invasive activity monitoring.

1. John Hasenau, DVM, DACLAM
Labanimalconsultants@charter.net
CEO and Principal Consultant,
Lab Animal Consultants
Stefano Gaburro, PhD
stefano.gaburro@tecniplast.it
Scientific Director, Digilab Division,
Tecniplast S.p.A.
Improving Animal Modeling with
24/7 Home Cage Monitoring in
Bioexclusion & Biocontainment
Mouse Housing Systems

2. Improving Animal Modeling with
24/7 Home Cage Monitoring in
Bioexclusion & Biocontainment
Mouse Housing Systems
Experts discuss current biosafety
requirements and what home cage
monitoring can teach us in bioexclusion
and biocontainment studies.

3. InsideScientific is an online educational
environment designed for life science
researchers. Our goal is to aid in the
sharing and distribution of scientific
information regarding innovative
technologies, protocols, research tools
and laboratory services

4. To access webinar content,
Q&A reports, FAQ documents,
and information on lab
workshops,
subscribe to our mail list

5. Bioexclusion and Biocontainment, are they
mutually exclusive with current rodent
housing systems?
Copyright 2020 J.Hasenau, Tecniplast and InsideScientific. All Rights Reserved.
John Hasenau, DVM DACLAM
CEO and Principal Consultant,
Lab Animal Consultants
E-mail: labanimalconsultants@charter.net

6. • How to accelerate bioexclusion and biocontainment research outcomes through
advanced caging systems.
• What are the practical differences in bioexclusion and biocontainment caging systems.
• Learn how locomotor activity serves as a “digital biomarker” for animal welfare for
infectious disease and metabolism studies.
• Understand how to improve the translational value of animal models via home
cage monitoring.
• Practical relevant information that you (the participants) can use at your facilities.
Key Objectives

7. 2018 
Integration
2015 – 2018
Paradigm Shift
2007 – 2015
Exploration
2005 – 2007
HMP
Launch
Timelines
Focus on Gnotobiotics - Bioexclusions

8. The Human Microbiome Project
An initiative of the NIH Roadmap for Biomedical Research
1st Phase of HMP (FY2007-2012) focused on the development of
metagenomics datasets and computational tools for
characterizing the microbiome in healthy adults and in cohorts of
specific microbiome-associated diseases
https://www.hmpdacc.org/
2007–2015: Exploration
The Human Microbiome Project

9. The Great Wave
Katsushika Hokusai (1760 –1849)
“Ology”
Gnotobiotics Microbiomics
With permission from Betty Theriault “Where is a mouse to live” FELASA 2019
What Changed in Research Paradigms?
“Omics”

10. September 3, 2018:
Pubmed Search “Microbiome” = 45,497 “hits” beginning in 1956
“Gnotobiotics” = 15,575 “hits” beginning in 1946
Narrow Range to 2005 – 2018 = 44,375 “Microbiome”
4,824 “Gnotobiotics”
Focus on Gnotobiotics - Bioexclusions

11. Trends in Publications for Gut Microbiota and Obesity

12. Mechanistic understanding of host-microbiome interactions
Wladarska et al, Cell 2014
Suez et al, Nature 2014
Thaiss et al, Cell 2014
Korem et al, Science 2015
Levy et al, Cell 2015
Zeevi et al, Cell 2015
Thaiss et al, Nature 2016
Gury et al, Cell 2016
Thaiss et al, Cell 2016
Korem et al, Cell Metab. 2017
Rothschild et al, Nature 2018
Thaiss et al, Science 2018
Suez et al, Cell 2018
Zmora et al, Cell 2018
Principles of microbiome-mediated disease treatment

13. Kennedy, Allergy Rhinol. 2020
Bassiouni et al, Infect Microbiol. 2020
Carney et al, Methods in Lung Microbiome
Research 2019
Hufnagl et al, Seminars in
Immunopathology 2020
Aguilera et al, Front. Immunol.
2020
Voskamp et al, Seminars in
Immunopathology , 2020
Bassiouni et al, Front. Cell. Infect.
Microbiol. 2020
Tan, Front. Cell Dev. Biol. 2020
Mechanistic understanding of host-microbiome interactions
Principles of microbiome-mediated disease treatment

14. Flexible Film Isolator
Present
Pre-1960
~1960
Germ Free Housing
Top photo depicts LOBUND Stainless Steel Isolators
Examples of early models of flexible film isolators
introduced by Philip Trexler at the LOBUND Institute.
The Laboratory Rat, Vol. 2 2006.
Hermetically sealed systems

15. Breeding and maintenance for long term housing of GF requires sterile conditions, which are easiest fulfill in
positive pressure isolators.
A GF breeding facility

16. Large isolators for breeding Wild Type mice Small isolators for maintaining Genetically
Manipulated GF mice strains
More Examples of GF breeding facilities

17. Development of the gut microbiome

18. Microbiota composition - Individual diversity

19. Our Measuring Tools
DNA Sequencing
RelativeAbundance Metagenome Metabolom

20. Our Tools
Healthy Healthy
Diseased Diseased
Germ-Free Germ-Free

21. 6 X 6
Up to 36
Microbiomes
Max of 4
Microbiomes
 Optimize Space
 Increase Resource
Availability
 At what Trade-offs?
Foot space Optimizations

22. Isocage use for Germ Free, Gnotobiotics and Biocontainment

23. Germ Free and Gnotobiotics in Isocages
Potential for using a hermetically-sealed, positive-
pressured isocage system for studies involving germ-
free mice outside a flexible-film isolator
Jisun Paik1,*, Olesya Pershutkina1
, Stacey Meeker1
, Jaehun JYi1
, SusanDowling1
, Charlie Hsu1
, Adeline M Hajjar1
, Lillian
Maggio-Price1
, and David A CBeck2,3
1The Department of Comparative Medicine; University of Washington; Seattle, WAUSA;2Department of Chemical Engineering; University of Washington; Seattle, WAUSA; 3
eScience Institute;
University of Washington; Seattle, WAUSA
Keywords: fecal transfer, germ-free, isolator, isopositive cage, microbiota, method
Abbreviations: MSD, multi-dimensional scaling; OTU, operational taxanomic unit; SOP, standard operating procedure.
Germ-free mice are used to examine questions about the role of the gut microbiota in development of diseases. Generally these animals
are maintained in semi-rigid or flexible-film isolators to ensure their continued sterility or, if colonized with specific microbiota, to ensure that
no new species are introduced. Here, we describe the use of a caging system in which individual cages are hermetically sealed and have their
own filtered positive airflow. This isopositive caging system requires less space and reduces animal housing costs. By using strict sterile
techniques, we kept mice germ-free in this caging system for 12 weeks. We also used this caging system and approach to conduct studies
evaluating a) the stability of the microbiome in germ-free mice receiving a fecal transplant and b) the stability of dietary-induced microbiota
changes in fecal-transplanted mice. As has been shown in fecal transfer studies in isolators, we found that the transferred microbiota
stabilizes as early as 2 weeks post transfer although recipient microbiota did not completely recapitulate those of the donors. Interestingly,
we also noted some sex effects in these studies indicating that the sex of recipients or donors may play a role in colonization of microbiota.
However, a larger study will be needed to determine what role, if any, sex plays in colonization of microbiota. Based on our studies, an
isopositive caging system may be utilized to test multiple donor samples for their effects on phenotypes of mice in both normal and disease
states even with limited available space forhousing.
Introduction
Studies utilizing germ-free mice are typically conducted in semi-
rigid or flexible film isolators designed to keep the animals
Intestinal commensal bacteria play significant roles in the in a sterile environment. To house and maintain animals under health and disease of
their host organisms, in part by participat- germ-free conditions, the introduction and removal of supplies ing in nutrient metabolism, guiding
proper immune system and samples to and from an isolator requires a time-consuming development, and influencing immune responses to
invading and labor-intensive decontamination process before and after organisms.1-6
Mounting evidence strongly associates particular opening of
the port to the isolator. If colonization experiments intestinal microbiota with diverse diseases, including obesity, need to be carried out using
multiple donor samples, each isola- autoimmune disease and inflammatory bowel disease.7
However, tor can house only a group of mice receiving a
single donor sam- due to the chronic nature of these diseases, it is difficult to deter- ple, requiring large space and expense. In addition,
experimental mine cause and effect. Hence, it is unclear whether changes in manipulation of mice in the isolator is cumbersome due to the the
complexity and structure of the microbiome are the precur- barrier and thick gloves worn by the experimenter to prevent sors or sequelae of these
diseases. Germ-free mice are a way to introduction of unwanted microbes.
address these questions as they provide an empty niche that can During the establishment of a gnotobiotic facility at our insti- be populated with
different microbiota, including those from tution, we developed standard operating procedures for the asep- human patients.5,8
Thus, germ-free
mice offer an opportunity to tic use of an alternative caging system to a flexible film isolator determine causality of certain bacterial communities
for a specific for studies using germ-free mice. We hoped that this system disease.7
would save time, space, and money while providing
ease of*Correspondence to: JisunPaik;Email:jpaik@uw.edu Submitted: 03/26/2015; Revised:
06/03/2015; Accepted: 06/15/2015 http://dx.doi.org/10.1080/19490976.2015.1064576
www.tandfonline.com Gut Microbes 255
Gut Microbes 6:4, 255--265; July/August 2015; ©2015 Taylor &Francis Group, LLC
RESEARCHPAPER

24. Gnoto Facility – Validated Experimental Procedures
Different types of
procedures can be
performed in the
ISOcage P or N
System.
Type of procedure
Cages
Specific diet provision
Specific water provision
Animals
Routine change
Feces collection
Body weight measurement
Body temperature measurement
Buccal swab
Intra-peritoneal injection
Gavage
Blood collection
Time-pregnancy mating
Surgery (vasectomy)
Surgery (renal ischemia-reperfusion)
Fetus transfer/ Axenization
Courtesy of Joana Bom,
Axenic/Gnotobiology Facility Manager

25. Iso Biosafety Station

26. Gnoto Facility – Experimental Diversity
Axenic Mice
. Sentinels
. Breedings
. New axenizations
. Experimental
Mono/bi colonized
Mice
. Experimental
Total fecal transplant
or Controlled
microbiota SOPF Mice
. ExperimentalCourtesy of Joana Bom,
Axenic/Gnotobiology Facility Manager

27. 4. ISOcage placed in the Gnotorack
until manipulation.
5. For animal manipulation on the IBS,
ISOcage undergoes a disinfection cycle.
1. ISOcage autoclaved
inside a cylinder.
3. ISOcage with axenic animals to
be transferred to the Gnotorack.2. Cylinder connected to the isolator to
remove animals.
7. Animals ready for manipulation,
or to be moved to Isolator trough an
autoclaved cylinder.
6. Inside the IBS, the cage is
removed from the dunk tank.
The Bioexclusion/Bioinclusion WorkflowAxenicGnoto
Courtesy of Joana Bom,
Axenic/Gnotobiology
Facility Manager

28. Decontamination Cart

29. Our tools
Healthy Healthy
Diseased Diseased
Germ-Free
or GEM
Germ-Free
or GEM
-Ve+Ve
Bio-Exclusion Bio-Containment

30. ISO-N Biocontainment studies

31. Keep the animals in their home cage: reduce animal and cage handling
Automated 24/7 data collection from the home cage-home provides unique advantages
DVC® for ISO
ResearchFacility Management /
Animal Welfare
 24/7 locomotor detection for a
better daily animal welfare
check
 Reduce the Bedding change
cage handling
 Automatic flooding detection
 24/7 locomotor data collection for more
reliable data
 High throughput data (all the experiment
cages together) minimize possible
confounding factors
 Provide standardized metrics: just keep the
animals in the home cage and automatically
collect results (reduced bias).

32. Thank you!

33. What can Home Cage Monitoring
Teach us in Biosafety Studies ?
Copyright 2020 S. Gaburro, Tecniplast and InsideScientific. All Rights Reserved.
Scientific Director, Digilab Division,
Tecniplast S.p.A.
Stefano Gaburro, PhD
E-mail: stefano.gaburro@tecniplast.it
Linkedin: https://www.linkedin.com/in/stefanogaburro

34. Mice spend 99% of their
time in the home cage.
More active at night
Most experiments
during daytime
Enviromental factors
(cage change, personnel)
are often
underconsidered
Cage change effects,
Weaning
effects longitudinal
studies (genetics*age)
Locomotion/Activity
behavioral test are
outside home cage
Only Snapshot,
Animal Welfare?
The experimental use of animals is evolving

35. Automated 24/7 data collection from the
home cage-home provides several advantages:
 Keep animals in their home cage:
reduce animal handling
 Allow animals monitoring during periods
generally not observed (e.g. nighttime)
 High throughput data: several available IVC
Cages running in parallel.
 Provide standardized metrics: just keep the
animals in the home cage and automatically
collect results (reduced bias).
Tecniplast Vision: Automated Home Cage Monitoring

36. DVC Structure
DVC® Rack
DVC® Interface
DVC® Server
Cloud
Locomotion(%)
Time (days)

37. DVC® Individual Mouse Tracking

38. DVC® in Science: Watch the E-Posters/Webinars

39. Empower your current ISOcage Units with the DVC® technology !!!
DVC® in Biocontainment Settings

40. Automated 24/7 data collection from the home cage-home provides unique advantages
DVC® for ISO
ResearchFacility Management /
Animal Welfare
Keep the animals in their home cage: reduce animal and cage handling
 24/7 locomotor detection for a
better daily animal welfare
check
 Reduce the Bedding change
cage handling
 Automatic flooding detection
 24/7 locomotor data collection for more
reliable data
 High throughput data (all the experiment
cages together) minimize possible
confounding factors
 Provide standardized metrics: just keep the
animals in the home cage and automatically
collect results (reduced bias).

41. Symptoms of Covid19
• Fever
• Cough
• Shortness of breath
• Tiredness

42. Infectious disease animal models: Covid19 model tested

43. 1. NHP and Larger
animals are
expensive
2. Ethical concern in
large animals or
NO SCALABILITY
Infectious disease animal models: Covid19 model...other considerations

44. Revisited Covid19 mouse model
Embryos cryopreserved @ Jax
Available Soon!
Why is ACE2 receptor important
for Covid19 entrance?
Animals kept 5 years,
then culled!

45. Revisited Covid19 mouse model

46. First publication on SARS-CoV-2 virus in hACE2 transgenic mice

47. Alternatives Covid19 mouse model (e.g. ACE2 Ko or BALB/C)

48. IVC*
What can we do with those models?
Isolators
Keep animals and visual inspection?
Measure sponteneous
locomotion in mice...
Why is it so important?
*Only permitted upon certain urgent conditions and by certain Nations
DVC®
ISO/N

49. Individual ventilated cages (IVC) in Covid19 Research

50. International Multicentric study including Jackson Lab (D)
C57BL6J – 4 animals/cage (female)
24/7 Locomotor Activity
Night
Day
Locomotoractivity
Age (Days)
Clear diurnal (day-night) locomotor activity differences!
What does a normal locomotor activity profile look like?

51. Virus (H1N1, Coronavirus, Spanish Flu), activity
and temperature acquired through minimiter
(implanted device)
Hypothermia (possibly lethargy) and loss of circadian locomotor activity
BODY TEMPERATURE LOCOMOTOR ACTIVITY
Viral effects of coronavirus on body temperature and activity

52. Control Vaccine Virus
LOCOMOTOR ACTIVITY
Balb/c mice (Venezuelan equine encephalitis virus)
Activity and Temperature through
Same activity
pattern of
vaccine treated
animal as for
control vs.
infected!!
Control Vaccine Virus
BODY TEMPERATURE
Vaccine effects monitored in the home cage

53.  Locomotor activity seems to demonstrate to
be predictive of virus pathological-induced effects
and its reversal by efficious vaccine
 DVC® has been used in the past by Jackson Labs to assess
locomotor activity in normal C57BL6/J mice
 Since the technology applied to biocontaiment is new, we lack
of data but we collected data from other pathological animals
models.
DVC® and locomotor activity in infectious disease

54.  SCID mice (n=216)
 Groups (cages) of 3-4-5 mice treated with cytostatic drug (cancer therapy) (200 mg/kg IP, 10mL/kg)
Body Weight
Hypoactivity
(-50%)
is detected prior
the onset of
body weight loss
(-3,5%)
Activity
Hypoactivity anticipates body weight reduction

55.  Locomotor activity demonstrates a great potential to additional
diagnostic tool in infectious disease as well to other experimental settings.
 DVC® might reduce needless cage opening and therefore minimizing viral
exposure.
 Core body temperature (implanted or rectal thermometer) or body
weight, at least in rodents, should be used in adjunction to locomotor
activity to assess reliably animal welfare in such study type.
Biocontainment Conclusions

56. Cryan et. Al, Lancet Neurology
Metabolism studies have gained major interest regarding dietary
effects on psychiatric disorders.
Many of such studies are performed in immunocompromised mice
(e.g. fecal microbial transplantation in left picture to study effects on
psychiatric disorders).
Surgery is performed in such animals and correctly assessing the
recovery represents an important aspect of animal welfare.
DVC® in Metabolism Research: Gnotobiotics

57. Total activity periphery duration is confirmatory of DVC® findings!!
Open FieldDVC® (day)
Confirmatory findings in Germ free animals: ADHD animal model

58. Confirmatory findings in Germ free animals: ADHD animal model

59.  DVC® can help to detect sick animals in infectious disease studies (e.g. COVID-19 article)
 DVC® can aid to detect and confirm locomotor activity patterns in germ-free or nude mice used
for gnotobiotics’ research
 Technology already validated (as for vaccine going to humans, e.g. Moderna- no animal
validation needed).
DVC® system for ISOcage units P and N represents a unique opportunity to empower these equipment and provide
useful 24/7 data for scientists in biocontainment and bioexclusion research fields.
Final Conclusions
For more information on Tecniplast solutions, please visit www.tecniplast.it or write to digilab-mktg@tecniplast.it

60. John Hasenau, DVM DACLAM
Labanimalconsultants@charter.net
CEO and Principal Consultant,
Lab Animal Consultants
Stefano Gaburro, PhD
stefano.gaburro@tecniplast.it
Scientific Director, Digilab Division,
Tecniplast S.p.A.
Thank You!
To learn more about digital housing solutiosn from
Tecniplast, please visit:
https://www.tecniplast.it/en/catalog/digilab.html