Sigma 3 18ks

The cultivated bacterial liquid was placed in a 100°C water bath for 10 min to inactivate the enzymes. The bacterial solution was centrifuged for 15 min at 4°C and 8,000 rpm. The supernatant was collected after centrifugation, 12% trichloroacetic acid solution of half the volume of the supernatant was added, and the proteins were removed from the supernatant after stirring for 30 min. Following centrifugation at 4°C at 8,000 rpm for 15 min (Sigma 3-18KS, Harzberg am Harz, Germany), the supernatant was collected, mixed with 2–3 times its volumes of 95% ethanol solution, and precipitated overnight. Then, the solution was centrifuged at 4°C and 8,000 rpm for 15 min to obtain a precipitate containing EPSs, after which it was weighed [11 (link),17 (link)].
Based on the quality of the EPSs, the precipitate was diluted with sterile water to prepare EPS solutions with different concentrations. After preparation, the bacteria were removed using a 0.22 µm filter (Millipore, MA, USA) and used as a backup (Table 1).

Cores were sectioned for pore-water collection in an anaerobic glove box (N₂ atmosphere with 3–5% H₂; Coy lab products, USA). Cores were sectioned at 0.5 cm resolution from 0 to 6 cm depth and 1 cm resolution between 6 and 12 cm depth. Sediment slices were collected in 50 ml polypropylene centrifuge tubes (TPP, Switzerland) and centrifuged at 3,000 rpm for 10 min (Sigma 3–18KS, Sigma Laborzentrifugen GmbH, Germany). Subsequently, the centrifuge tubes were opened in the anaerobic glove box, and overlying pore water was transferred into suitable sample containers after filtration through 0.45 μm cellulose filters (Millex-HA filter, Merck Millipore, USA). Pore-water samples were analyzed for dissolved iron and dissolved As. To prevent oxidation, the solid phase that remained after centrifugation was freeze-dried and sealed in an airtight aluminum bag inside the anaerobic glove box and stored anaerobically for later solid-phase analysis.

First, a 4% sodium alginate solution was prepared from distilled water and alginic acid sodium salt. It was used throughout the experiment for emulsion preparation as the shell material. Emulsion with Trifolium pratense L. and Glycyrrhiza glabra L. extracts, and Myristica fragrans Houtt. essential oil were prepared as follows: solution with excipients (maltodextrin, inulin, and/ or gum Arabic) was mixed with sodium alginate solution (stirred for 15 min with a magnetic stirrer MSH-20A (Witeg, Wertheim, Germany)) and then extracts with essential oil were added. The solution was homogenised for 15 min at 5000 rpm using an IKA T18 homogeniser (IKA-Werke GmbH & Co., KG, Staufen, Germany).
The emulsion’s stability was tested using a centrifuge Sigma 3-18KS (Sigma Laborzentrifugen GmbH, Osterode am Harz, Germany). The test was repeated three times using 23 °C temperature, 3000 rpm, and the duration was 5 min. The centrifugation index (CI) was calculated to evaluate emulsion stability.
$$\mathrm{CI}(\%)=\frac{{\mathrm{V}}_{\mathrm{e}}}{{\mathrm{V}}_{\mathrm{i}}}·100$$
where V_e is the volume of the remaining emulsion after centrifugation and V_i is the volume of the initial emulsion.

First, 4% sodium alginate solution was prepared from distilled water and alginic acid sodium salt. It was used throughout the experiment for emulsion preparation as the shell material. Emulsion with polysorbate 80 was prepared as follows: 4% sodium alginate solution (5 g, 10 g, and 15 g) was diluted with distilled water (10 mL, 5 mL, 0 mL), a surfactant (0.5 g, 1 g, 1.5 g) was added, everything was stirred with a magnetic stirrer MSH-20A (Witeg, Germany) for 10 min and then the nutmeg essential oil (0.5 g, 1 g, 1.5 g) was added. The emulsion was stirred for 15 min. The absorbent magnesium aluminometasilicate (0.2 g and 0.4 g) was used in several samples; this material was added before the essential oil.
The emulsion with Sisterna SP70 was prepared by three different methods (Figure 1).
The stability of each emulsion was tested using a centrifuge Sigma 3-18KS (Sigma Laborzentrifugen GmbH, Osterode am Harz, Germany). The test was carried out in these conditions: 3000 rpm, temperature 23 °C, and duration 5 min. All samples were tested three times.

Animals were anesthetized
using isoflurane, and blood samples (1 mL) were collected from the
lateral tail vein in 1.5 mL autocued plastic tubes between 9:00 am
and 12:00 pm (dark cycle). The samples were allowed to stand for 10
min before centrifuging (Sigma 3-18KS, Germany) at 3000 rpm (800g) for 10 min at 23 °C, after which serum was collected
into duplicate 0.5 mL autocued plastic tubes and stored at −80
°C until assay.

Pollen of Egyptian date palm (Phoenix dactylifera L.) were harvested at the end of the March 2021 and 2022 seasons at the beginning of opening covers of the male species from Rashed city, Elbehyra Governorate, Egypt. The extract of pollen was prepared according to the authors of [95 (link)], with some modifications, as follows: to prepare the water pollen extract, 0.1 g of pollen grains was added to 10.0 mL of distilled water. After one hour, the mixture was sonicated using a VCX 750 ultrasonic probe (SONICS & MATERIALS, INC., Newtown, CT, USA) (frequency of 6 kHz), cut for 30 s, and then centrifuged (Sigma 3–18 KS, SIGMA Laborzentrifugen GmbH, Osterode am Harz, Germany) at 5000 rpm for 10 m in at a temperature of 20 °C. The resulting supernatant was used as the water pollen extract in all the experiments. Then, the volume was completed with water to obtain the used concentrations (10 and 20 g/L) [56 ]. The tested bio-stimulant was applied as a foliar spray thrice. The 1st and 2nd sprays were carried out after transplanting at 32 and 62 days, respectively, and the third one was conducted 22 days after the first cut. For the NPs and DPPE, tween 80 was applied as a sticking agent in each spray. The spraying was carried out with a hand sprayer in the morning. The plants were sprayed till the spray ran off.