Severity assessment in mice subjected to carbon tetrachloride

Abstract

The Directive 2010/63 EU requires classifying burden and severity in all procedures using laboratory animals. This study evaluated the severity of liver fibrosis induction by intraperitoneal carbon tetrachloride (CCl4) injections in mice. 29 male C57BL/6N mice were treated three times per week for 4 weeks with an intraperitoneal injection (50 µl) of either 0.6 ml/kg body weight CCl4-vehicle solution, germ oil (vehicle-control) or handling only. Severity assessment was performed using serum analysis, behavioral tests (open field test, rotarod, burrowing and nesting behavior), fecal corticosterone metabolite (FCM) measurement, and survival. The most significant group differences were noticed in the second week of treatment when the highest AST (1463 ± 1404 vs. 123.8 ± 93 U/L, p < 0.0001) and nesting values were measured. In addition, respective animals showed lower moving distances (4622 ± 1577 vs. 6157 ± 2060 cm, p < 0.01) and velocity in the Open field, identified as main factors in principal component analysis (PCA). Overall, a 50% survival rate was observed within the treatment group, in which the open field performance was a good tracer parameter for survival. In summary, this study demonstrates the feasibility of assessing severity in mice using behavioral tests and highlight the open field test as a possible threshold parameter for risk assessment of mortality.

Closing an Eye for the Reliability of Orbital Tightening in Pain Detection: A Model-Specific Simplification of the Mouse Grimace Scale

“…This indicates a painful impulse caused by the injection, which lasts over the investigation period of one hour after injection. These ndings are in line with our recently published study on the severity of the CCl 4 model itself 24 , which showed the highest severity of the animals in various clinical and behavioral parameters also during the second week of treatment. In Fig.…”

“…However, this habituation effect in the CCl 4 group may be due to the increased liver metabolism in the turnover of toxic CCl 4 with the second week of treatment. These changes in liver metabolism were shown elsewhere by blood analysis in the CCl 4 model 24 .…”

“…The study protocol complied with the EU Directive 2010/63 and the Guide for the Care and Use of Laboratory Animals 25 . This study was performed in accordance with the application of the 3Rs criteria as a branch project from a recently published animal study on evaluation severity assessment in brosis induction 24 . The animals were examined retrospectively, no additional experiments were carried out.…”

“…These animals were weighed and then divided into two treatment groups: A CCl 4 group and a control group (oil) for further investigation in a liver brosis model. 24 . For this purpose, the animals were injected i.p.…”

L-glutamic acid effect on changes in biochemical parameters of rats intoxicated by carbon tetrachloride

“…Ключові слова: L-глутамінова кислота, щурі, кров, CCl 4 , АлАТ, АсАТ Потенційна небезпека для організму ссавців від впливу техногенних чинників зумовлена, зокрема, активацією вільнорадикальних реакцій, виникненням тканинної гіпоксії та порушенням детоксикаційної функції печінки. Тетрахлорметан (CCl 4 ) -токсична органічна речовина штучного походження, добре відома у науковій практиці як модельна сполука для вивчення ураження паренхіматозних клітин печінки [7,10,12,13]. За структурою це хлорований вуглеводень.…”