FarmBioLine Oy и ООО ”ФармБиоЛайн”
Семинар:
«Использование ИВК систем для содержания и работы с лабораторными
животными. Оборудование компании Ugo Basile для научных исследований»
Использование систем ИВК:
выводы по результатам фенотипирования
лабораторных животных
ИЮ Зарайская
[email protected]
22-го Октября 2014 г.
factor that might affect neural and behavioral phenotypes
air renewal
ventilation system noise
chronic stressors induce major changes
in behavior and neurobiology
subtle vibrations of the racks
olfactory and acoustic
interchange cues limitation
restricted sensory input from
the outside
IVC
housing
standardization of mouse
handling & housing conditions:
minor changes in procedures
profoundly affect biological
variables
frequency of cage changes
IVCs design can vary markedly
behavioral testing in an open air environment
complicating replication of
previously obtained data or biasing
ongoing studies
Experimental Protocols
IVC housing vs. IVC housing
IVC housing vs. Conventional housing
pre-experimental IVC housing vs. pre-experimental FILTER housing
pre-experimental IVC housing vs. pre-experimental Conventional housing
Experimental Animals
Mice
Female C57BL/6J, Charles River, Italy
Female BALB/c, Charles River, Italy
male & female C57BL6/Jarc, Australian
BioResources
C3HeB/FeJ, German Mouse Clinic
C57BL/6J, German Mouse Clinic
males & females BALB/cJ, C57BL/6J, DBA/2J;
Jackson Lab, USA
IVC Systems
Techniplast, Italy
Allentown, USA
BioZone, UK
IVC M.I.C.E.® (Animal Care System)
Innocage® (Innovive Inc.).
Rats
Slc6a41Hubr - serotonin transporter
knock-out rats; F10 out-crossed
(Harlan, Netherlands)
Behavioral Protocols
Indirect Behavioural
& Physical Measures
•Injury/wound scores
•Body hair removal
•Whisker-trimming
•Bedding pushing
•Bodyweight (g)
•Water utilisation (ml)
•Food utilisation (g)
•Saccharin preference
Within-cage behaviour
•Animal location
•Movement by biotelemetry
Faecal corticosterone
Behavioral Tasks
•Spontaneous locomotor activity
•Open Field
•Modified hole board
•Elevated Plus Maze
•Forced swim test
•Y maze
•The 8-arm radial maze
•Novel object recognition
•Social interaction
•Resident intruder test
•Рrepulse inhibition
•The acoustic startle reflex
•Fear potentiated startle
•Fear conditioning
•Fear extinction
•Baseline and following acute
treatment with NMDA antagonist
MK-801
•Sickness behavior - IP 100
μg/kg lipopolysaccharide (LPS)
2007
IVC housing vs. Conventional housing
(BioZone, UK)
C3HeB/FeJ, German Mouse Clinic
C57BL/6J, German Mouse Clinic
•Modified hole board (mHB).
•Acoustic startle reflex (ASR)
•Fear potentiated startle (FPS)
Modified Hole Board
housing: F(2,56) = 5.07, p < 0.01
housing:
x2(6)
= 28.09, p < 0.001
housing: F(2,56) = 9.27, p < 0.001
housing: F(2,56) = 8.38, p < 0.001
housing: LR x2(4) = 15.07, p < 0.01
housing: LR x2(6) = 19.92, p < 0.01
housing: LR v2(6) = 16.55, p < 0.05
Conclusions
•The present findings show that IVC housing can affect behavioral performance.
•The behavioral effects of IVC housing strongly dependent on genetic
background of experimental subjects.
2009
IVC housing vs. Conventional housing
Allentown Caging Equipment Co, NJ, USA model V1944-09-00
males & females BALB/cJ, C57BL/6J, DBA/2J; Jackson Lab, USA
Plus maze
Open field
Radial maze
Resident intruder test
Startle reflex
Plus maze
The percentage of time spent
in the open arm:
housing effect (F1, 171=0.052, p=0.82)
strain effect (F2, 171=13.98, p<0.0001)
the housing effect was different across sex
and strains (sex X strain X housing:
F2, 171=5.23, p=0.0064)
The total number of arm entries:
housing effect (F1, 171=2.55, p=0.11)
strain effect (F2, 171=152.22, p<0.0001)
Sex X strain X housing interaction
(F2, 171=3.79, p=0.024)
Open field
strain X housing: F2, 158=4.52, p=0.012
(strain X housing: F1,158=7.87, p=0.0006)
Sex X strain X housing: F2, 158=6.02, p=0.003
Resident-intruder test
strain X housing (F2, 74=4.74, p=0.01)
housing X strain (F2, 74=4.62, p=0.012)
Startle reflex
Conclusions
• IVC housing does not have a clear and unequivocal effect on the behavior
of mice.
• The behavioral effects of caging environment turn out to be strongly
dependent on strain and sex of the experimental subjects.
2014
IVC housing vs. IVC housing
(Techniplast)
(Allentown)
Female C57BL/6J, Charles River, Italy
Female BALB/c, Charles River, Italy
• Measures of general behavior and anxiety-related behavior
Open field
Plus maze
‘internal’ crossing frequency:
Strain ∗ System effect (F1,28 = 7.7, P = 0.01)
defecation:
System effect (F1,28 = 12.2, P = 0.002)
time in the internal/central zone:
System effect (F1,28 = 6.2, P = 0.019)
time in the open arm:
System effect (F1,28 = 18.5, P = 0.001)
grooming duration:
System effect (F1,28 = 8.2, P = 0.008)
time in the centre:
System effect (F1,28 = 13.6, P = 0.001)
Faecal Corticosterone
Conclusions
•There are significant differences between IVC systems in their influence on mouse
behavior and anxiety
•The results indicate that IVC systems that vary in air delivery features also appear
to influence anxiety-related behavior.
•IVC housing System Two (with air delivery at the ‘animal’ level at 50 ACH) was more
anxiety-inducing than IVC housing System One, with air delivery at the ‘cover’ level
(and at 75 ACH), for mice of both strains (BALB/c, C57BL/6J).
2012
pre-experimental IVC housing vs. pre-experimental Conventional housing
Micro-VENT (IVC) system (Allentown)
C57BL/6J, Jackson Lab, USA
Social exploration
Novel object recognition
Spontaneous locomotor activity
Results
IVC mice have an increased preference for saccharin but no change in
locomotion, social exploration or novel object recognition
2013
pre-experimental IVC housing vs. pre-experimental FILTER housing
Airlaw, Smithfield, Australia
male & female C57BL6/Jarc, Australian BioResources
•elevated plus maze
•Y maze,
•fear conditioning
•prepulse inhibition
Plus maze
* - significant level
for factor “Housing”
* - significant level
for factor “Housing”
* - significant level
for factor “Housing”
Conclusions
•In the current study IVC systems had no impact on locomotion and exploration of male
and female C57BL/6J mice in a variety of test paradigms
•Female mice kept in IVC developed an increased anxiety response compared to FILTERhoused females.
•Testing mice in cognitive tasks for cue and contextual FC and short-term recognition
memory revealed no impact of housing conditions or sex on relevant learning
and memory processes.
•Sensorimotor gating and the acoustic startle response of male and female test mice were
not affected by the two housing conditions.
заключение
Различия в поведении, наблюдаемые в условиях конвенционального и ИВК- содержания,
зависят от генотипа и пола.
Наиболее чувствительным к условиям содержания является эмоциональный профиль
животных, в частности, уровень тревожности.
Условия содержания (конвенциональное, тип ИВК систем) могут взаимодействовать с
генетическими, фармакологическими, или другими манипуляциями.
Термин «ИВК» не может быть «генерализован» на все типы ИВК-систем. Их вариации могут
определять различия в поведении и, соответственно, в получаемых данных.
К сожалению, большинство экспериментальных поведенческих данных на грызунах не
указывают, какой тип систем был использован. Это делает проблематичным возможность
сравнения результатов тестирования между лабораториями и т.д.
Условия выращивания и содержания могут изменяться. Исследователь должен понимать,
что это может привести к изменению получаемых им экспериментальных данных.
Идентифицированные однажды факторы выращивания и содержания должны приниматься
в расчет экспериментального дизайна и дальнейшего статистического анализа.
Различия в условиях выращивания и содержания в питомниках и лабораториях должны
обсуждаться. Экспериментальная работа должна быть тщательно продумана.
Tecniplast IVCs compliant with 8th edition of the NRC
Guide for the Care and Use of Laboratory Animals
The 8th (2011) edition of the Guide for the Care and Use of Laboratory Animals (1) , published by the National
Research Council of the National Academies, discusses issues related to the speed, rate and supply of air in
individually ventilated cage systems ( IVCs). The possible impact of the direct exposure of animals to air
moving at high velocity (=0.3m/sec) is a specific concern.
The Guide states on page 46 “ ..direct exposure to air moving at high velocity should be avoided as the
speed of the air to which animals are exposed affects the rate at which heat and moisture are removed
from an animal”.
This issue has been raised by a number of authors who cited the high turnover of air as a possible cause of a
stressful, dry, micro-environment (2, 3, 4, 5).
In 2002, Baumans et al (6), in a study of commercial IVC systems, concluded “we found that mice prefer
having large cages and an air supply in the cover, at least in the studies systems, and nesting material
should be supplied whenever possible”.
Discussing the effects of draughts, Krohn & Hansen (7) concluded that: ”the point of air entrance into the
cage can be responsible (due to the high air speed directed at the animal) of welfare problems in mice
and consequently the air should not be blown into the cage at animal level, irrespective of the number
of air changes”.
Copyright Tecniplast Group 2014
Клетки с индивидуальной вентиляцией
Крышка с фильтром
Самозакрывающийся воздушный клапан
обеспечивает целостность пространства и
проникновение воздуха в каждую клетку при низкой
скорости (< 0,2 м/с)*.
Поилка располагается в
углублении и может
быть заменена без снятия клетки с
IVC стеллажа, что позволяет
минимизировать количество
открываний клетки.
литой нейлоновый замок и
фиксаторы обеспечивают
надежное крепление крышки к
клетке
Воздух фильтруется через НЕРА фильтр и попадает в клетку на
низкой скорости через самозакрывающийся клапан,
подключенный к системе вентиляции стеллажей.
Диффузор клапана клетки и направляющая перегородка
направляют воздух вдоль верха к передней части клетки и вниз,
создавая легкий обратный поток на уровне подстилки по
направлению к вытяжке.
Прямая вытяжка из клетки увеличивает эффективность
вентиляции, в результате чего контролируется температура и
влажность. Благодаря этому подстилка остается суше и снижается
уровень аммиака, что позволяет увеличить интервал между
заменами подстилки, без необходимости вентиляции на уровне
животных
Фильтр 0,2 μ PALL для защиты от бактерий и аэрозольных частиц
ограничивает воздействие CO2 при отсутствии вентиляции без
необходимости применения запасной батареи питания
Science. 1975 Aug 8;189(4201):465-7.
Mouse mammary tumors: alteration of incidence as apparent function of stress.
Riley V.

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