Digesta samples were collected from the rumen mat for bacterial 16S rRNA gene Illumina MiSeq (Illumina, Balgach, Switzerland) sequencing, and samples of particle-associated rumen liquid were obtained for measuring short-chain fatty acids, lactate, ammonia, and pH during RD (d 6), Conc I (d 19), and Conc II (d 39).
Totally, nine sets of samples were obtained for measuring the dry weight of superior and inferior spikelets, respectively.
Daily samples of arterial blood and venous jugular bulb blood were obtained for measuring pCO2, lactate, serum sodium and CRP.
After a brief centrifugation at 1600 × g for 10 min at 4°C, the supernatants were obtained for measuring absorbance at 540 nm.
Blood sample and cervical exfoliated cells were obtained for measuring serum retinol, α-carotene, β-carotene, zeaxanthin/lutein, cryptoxanthin, lycopene and α-tocopherol and for HPV detection.
A venous blood sample was obtained for measuring lipids, glycerol, FAs, albumin, sensitive CRP and IL-6, as previously described (Agustsson et al, 2007; Ryden et al, 2008).
On arrival, and after signing a consent form, a fasting blood analysis will be obtained for measuring the blood levels for glucose, creatinine, glycated haemoglobin (HbA1c), lipids and lipoproteins.
Whole blood and urine samples were obtained for measuring the serum creatinine, serum cystatin C, and urine and plasma NGAL levels at 0, 3, 6, 12, 24, and 48 hours after admission.
BALF and lung tissue were obtained for measuring levels of complement activation (C4b/c in BALF), lung injury (total protein levels in BALF), and inflammation (IL-6 levels in lung tissue).
Furthermore, no reliability or validity data were obtained for measures of ideation, plan or attempt, restricting inferences that can be drawn from this study.
