Experimental protocol over the 2 generations

Twenty-eight pregnant New-Zealand white female rabbits (INRA1077 line, 1-year old) (F0) were exposed by nose-only inhalation in custom made plexiglas tubes to either diluted DE (1 mg/m3) (exposed group) or clean air (control group) for 2 h/day, 5 days/week, from the 3rd to the 27th day post-conception (dpc) (i.e., 20 days altogether over a 31-day gestation) (Fig. 1). DE exposure was performed with the Mobile Ambient Particle Concentrator Exposure Laboratory [18 (link)] connected to a 25KVA Loxam engine, with a 500 nm particle filter (Additional file 1: Figure S1).
DE is a complex mixture of hundreds of constituents in either a gas or particle form. Gaseous components of DE include carbon dioxide, oxygen, nitrogen, water vapour, carbon monoxide, nitrogen compounds, sulphur compounds, and numerous low-molecular-weight hydrocarbons (some of them individually known to be toxic, such as aldehydes, benzene, 1,3-butadiene, polycyclic aromatic hydrocarbons (PAHs) and nitro-PAHS). The particles present in DE are known to be composed of center core of elemental carbon with absorbed organic compounds and small amounts of sulphate, nitrate, metals, and other trace elements [19 ]. The measured components of the exposure mixture in the present experiment are shown in Additional file 2: Table S1.
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