Falcon tube

In this experiment exclusively, Pi and ${\text{NO}}_3^{-}$ concentrations were monitored in the Hoagland^dil100X solution of the M and NM plants grown in the S-H cultivation system. Twenty ml of Hoagland^dil100X solution was sampled from each bottle after each circulation (T1, T2, T3, and T4). At each sampling time, the solution was collected in 50-ml Falcon Tubes (Sarstedt, Germany) and then stored at 4°C in the dark before proceeding with the analysis.
Concentration of Pi was evaluated via inductively coupled plasma atomic emission spectroscopy (ICP-AES) as described by Garcés-Ruiz et al. (2017 (link)). For ${\text{NO}}_3^{-}$ , samples were first diluted 50 times with Milli-Q water and then analyzed via ionic chromatography system (IC, DIONEX, DX 120). The ${\text{NO}}_3^{-}$ quantification was analyzed under ultraviolet-visible (UV/VIS) spectrophotometry detection between 200 and 220 nm. The limit of detection was <100 ppb.

Root colonization was estimated on three replicates from the PM and NM plantlets before experiments set up and all the PM replicates at the end of the experiment (i.e., at 17 and 46 days, respectively). Roots were sampled and analyzed via ink staining (Walker, 2005 ). The roots were cut into small pieces and placed in Falcon tubes (Sarstedt, Germany). Twenty-five milliliter of KOH 10% was added to the roots before incubation at 70°C in a water bath for 1 h. The KOH was then removed and roots washed with HCl 1%. The staining step consisted in adding 25 mL of ink 2% (Parker blue ink, United States) in HCl 1%. The tubes were then placed at 70°C in a water bath for 1 h. The roots were rinsed and stored in deionized water before observation (Walker, 2005 ). Total (TC), arbuscular (AC) and vesicular (VC) colonization were estimated under a dissecting microscope (Olympus BH2–RFCA, Japan) at 10× magnification (McGonigle et al., 1990 (link)). Around 170 intersections were observed per plantlet. At each intersection the presence or absence of the fungus was noted.

Fecal samples for the analysis of cortisol metabolites 11,17-dioxoandrostanes (11,17-DOA) concentrations were collected rectally from each animal between 7:30 a.m. and 10:30 a.m. The samples were stored in 50-ml falcon tubes (Sarstedt, Nürnbrecht, Germany), immediately cooled by freezer packs, and within 3 h after collection frozen at −20°C until assay.
The frozen fecal samples were thawed for 30 min at room temperature. The 0.5-g wet feces of each sample was vortexed on a multi-vortex (VWR International GmbH, Darmstadt, Germany; Tube Rotator) for 30 min with 5-ml 80% methanol (Avantor, Griesheim, Germany; J.T. Baker^® Chemicals) and centrifuged at 2,500 × g for 15 min at room temperature. The supernatant was pipetted into a tube and stored at −20°C until analysis. 11,17-DOA was analyzed using the group-specific 11-oxoetiocholanolone enzyme immunoassay (EIA) from Möstl and Palme (Labcode 72a, Department of Biomedical Sciences, Physiology, University of Veterinary Medicine, Vienna, Austria). Extraction and EIA were carried out as described by Palme and Möstl (38 ). Statistical analysis of 11,17-DOA concentration was carried out for days 2, 3, and 5 of each experimental period (B, T1, T2, T3).

To investigate if the brine microbiota could lead to biogenic amine production, small-scale fermentation processes with an inoculum of 1% (vol/vol) brine 1 (brine sample B) were performed in sterile, closed 15 mL Falcon tubes (Sarstedt, Nümbrecht, Germany) at 15°C. Therefore, two different media were used. A liquid medium optimal for the production of biogenic amines was prepared as defined before (81 (link)) but without meat extract and supplemented with 5 g/L of NaCl. Also, a cheese medium as described above was used. Samples were taken after 0, 1, 2, and 4 weeks of fermentation, and biogenic amines were quantified as described previously (21 (link)).

The hunters were instructed to collect samples from shot wild boars including tonsils, one mesenteric lymph node, and faeces. The samples were frozen in 15 mL Falcon tubes (Sarstedt AG & Co, Nümbrecht, Germany) and sent on ice by ordinary mail to the laboratory. All samples were kept frozen at − 20 °C until analysis (maximum 18 months storage). The questionnaire was to be filled out and sent in with the samples and included questions on sex, weight and time of sampling of each wild boar, and information on the population characteristics for the population in the area where the wild boars were shot.
The questions on population characteristics [24 ] are given in Table 1.Table 1

The distribution of the 30 wild boar populations in the respective risk factor category, based on the answers in a questionnaire that accompanied the samples

Risk factors	Number (%) of populations in each category
Feeding intensity (feeding places/10 km2)
< 3	7 (23.3)
3–5	15 (50.0)
5–10	3 (10.0)
> 10	5 (16.7)
Years since establishment of population (years)
< 3	1 (3.3)
3–5	3 (10.0)
5–7	4 (13.3)
7–10	5 (16.7)
> 10	17 (56.7)
Yearly harvest/10 km2 (animals)
< 5	6 (20.0)
5–15	12 (40.0)
15–30	3 (10.0)
30–50	5 (16.7)
> 50	4 (13.3)
Handling of slaughter waste
Made unavailable for wild boars	11 (36.7)
Left out in the forest	19 (63.3)

Organ transport and perfusion fluid collected at time of liver transplant was centrifuged in 50 ml falcon tubes (Sarstedt) at 1,800 rpm for 15 min and the supernatant discarded. The tubes were vortexed to disrupt the pellet and the cells pooled and resuspended in RPMI before density gradient centrifugation over Ficoll Hypaque as above.

The embryos were checked for viability before exposing them to either 18°C, 23°C, 28°C, 33°C, or 37°C in sealed 50ml falcon tubes (62.547.254, Sarstedt, Germany) in a temperature-controlled water bath. Up to 70 embryos were transferred into the falcon tubes containing 50ml of E3 medium. The embryos were exposed to the respective temperature for 20h (see Supplementary Figure). We did not term temperature exposure “acclimation”, as we did not evaluate acclimation steady states. At 48 hpf, the zebrafish embryos were transferred to respirometric analysis.