Ad was labeled by injections of anterograde tracer (Alexa 488 conjugated dextran 10 MW) into the nidopallium lateral to LMAN (LMAN-shell), and AIV was retrogradely labeled in the same birds by injection of CTB into VTA and SNc (n = 8 hemispheres).
On the evening of 12th March after the lights had been switched off, four birds received an injection of 0.1 mg E. coli LPS (strain 055 B5, Sigma L2880) in 40 μL sterile isotonic saline into the pectoralis muscle.
Positive antibody controls for HI and MN assays were prepared by immunization of birds with s.c. injection of an emulsion containing 0.5 mg of whole, formaldehyde-inactivated influenza NIBRG-14 four times with complete or incomplete Freund's adjuvant (Calbiochem, La Jolla, CA, USA).
Like the coal mines, which were often named to conjure up riches, fantasy and benevolence – the Century mine, the Freedom mine, the Rosebud mine – the injection wells of Appalachia have misleading, lovely names: Flowers well, Red Bird Injection well, Hazel Ginsburg.
Following 10 minutes of its circulation, the bird was killed by injection of 5 ml of thiopentone sodium into the right brachial vein.
On the morning of the following day, a blood sample was obtained from each bird using a pre-heparinized needle and syringe and the birds were killed by intravenous injection of sodium pentobarbitone.
For tracer injections, birds were anaesthetized by intramuscular injection of ketamine (Pfitzer, Freiburg, Germany)/rompun (Bayer, Leverkusen, Germany), and their heads were fixed in a custom-built stereotactic apparatus.
Two birds were killed by intramuscular injection of an overdose of Narcoren (Merial, Hallbergmoos, Germany).
All birds were euthanized by intravenous injection of a barbiturate overdose followed by decapitation.
BiRD was stimulated by direct injection of Epson salt (MgSO4·7H2O) and sodium (L) lactate (NaC3H5O3) in five injection wells.
