Protocols for field and laboratory rodent studies - HAL

ContentsForeword ............................................................................................................................................................... IIIBackground: CERoPath project ........................................................................................................................... VContents ................................................................................................................................................................. 11. Trapping rodents .......................................................................................................................................... 31.1. Trapping techniques ............................................................................................................................. 31.2. Trapping protocol .................................................................................................................................. 41.3. Recording trapping data ........................................................................................................................ 52. Dissecting rodents ....................................................................................................................................... 62.1. Organization of the field laboratory ....................................................................................................... 62.1.1. Processing site .............................................................................................................................. 62.1.2. Avoid handling live animals ........................................................................................................... 62.1.3. Preparation of tubes ...................................................................................................................... 62.2. Euthanasia ............................................................................................................................................ 72.3. Collection of ectoparasites .................................................................................................................... 92.4. Blood collection ................................................................................................................................... 102.4.1. Blood collection from the base of the tail ..................................................................................... 102.4.2. Blood collection from the orbital sinus (retro-orbital).................................................................... 102.4.3. Blood collection by cardiac puncture ........................................................................................... 112.4.4. Blood collection from a dead rodent. ........................................................................................... 112.4.5. Serum separation and blood storage ........................................................................................... 112.5. Dissection ........................................................................................................................................... 142.5.1. Separation of organs ................................................................................................................... 142.5.2. Hemoparasites: detection, identification and cryopreservation .................................................... 182.5.3. Leptospira: culture preparation from kidneys ............................................................................... 192.5.4. Orthopoxviruses: culture preparation from urine and tracheal mucus. ......................................... 193. Identifying rodents ..................................................................................................................................... 203.1. External identification .......................................................................................................................... 203.1.1. External measurements ............................................................................................................... 203.1.2. Sex determination ........................................................................................................................ 223.1.3. Maturity determination and development stage ........................................................................... 223.1.4. Data recording and species identification .................................................................................... 233.2. Morphometrics .................................................................................................................................... 243.3. Genetics .............................................................................................................................................. 253.3.1. Species-specific PCR-tests.......................................................................................................... 253.3.2. DNA barcoding ............................................................................................................................ 273.4. Cytogenetics: karyotyping and tissue sampling for fibroblast cell culture............................................ 293.4.1. Karyotypes: mitotic chromosome preparation from bone marrow ................................................ 293.4.2. Tissue sampling for cell culture.................................................................................................... 314. Isolation of helminths from rodents ......................................................................................................... 324.1. Gastrointestinal (GI) tract and helminths isolation .............................................................................. 324.2. Helminths collection in the gastrointestinal tract ................................................................................. 344.3. Method for karyological analysis of helminths ..................................................................................... 354.4. Helminths fixation for scanning electron microscopy .......................................................................... 355. Preparing rodent specimens for collections ............................................................................................ 365.1. Dry collections .................................................................................................................................... 365.1.1. Skin preparation ........................................................................................................................... 365.1.2. Skull preparation .......................................................................................................................... 375.2. Wet collections .................................................................................................................................... 385.3. Data sharing with online collections .................................................................................................... 38References............................................................................................................................................................ 39Other relevant guidelines .................................................................................................................................... 39Glossary ............................................................................................................................................................... 40List of figures ....................................................................................................................................................... 42List of tables ......................................................................................................................................................... 43Appendixes........................................................................................................................................................... 431