Sl16r

Five different techniques were tested such as (i) freeze drying (FD); (ii) air drying (AD); (iii) sun drying (SD); (iv) oven drying (OD) and (v) microwave drying (MD). For that purpose, 1 L sample of T. subcordiformis culture was collected from each tank on the same day for each technique to be tested. The drying was performed in triplicate using 1 L from each tank of 20 L (1 L sample per tank, and three replicates per drying technique). After harvesting using a cold centrifuge (SL16R, Thermo Fisher), the wet biomass was spread into a glass plate prior to being dried via the following techniques:

Freeze drying (FD): After an ON incubation at − 80 °C, the biomass was transferred into a freeze dryer (Labconco, 7754067) for a duration of 48 h,

Sun drying (SD): wet biomass was incubated directly under the sun for 48 h in a location that allows the maximum of illumination hours,

Air drying (AD): biomass was covered with 4 layers of aluminum and incubated at room temperature for 48 h,

Microwaves drying (MD): wet biomass was incubated in a microwave (Panasonic NN-CD9975) for a duration of 2 h with a successive incubation of 15 min until completely dry.

Oven drying (OD): the biomass was incubated for 48 h in an oven (Binder EP-53) under 90 °C.

Treatments were performed in parallel and stopped after confirming a stable weight of the dry biomass.

Based on the results of the viability test, cells were exposed to KB (βHB 4 mM, AA 1 mM) for 2, 24 and 48 h at 37 °C with 95% humidity and 5% CO₂. Every treatment was performed in triplicate; negative (cells without KB) and positive (cells treated with H₂O₂: 50 μM for 5 min) controls were included in each batch. After incubation, an aliquot of cells was used to verify cell viability via the trypan blue exclusion test [38 (link)], to be sure that cells were suitable for the comet assay. Another aliquot of cells was centrifuged (11,000× g, for 15 s; SL16R Thermo Fisher Scientific), re-suspended in 1.5% low melting point agarose, and spread on a microscope slide previously covered with 1% normal melting point agarose. Embedded cells were lysed, DNA was allowed to unwind in electrophoresis buffer (pH 10) and then electrophoresis was performed at 25 V and 300 mA for 20 min. The slides were then immersed in neutralization buffer for 15 min, stained with ethidium bromide and analyzed using a fluorescence microscope (BX60 Olympus, Tokyo, Japan) equipped with Image-Pro Plus software (Immagini & Computer, Bareggio; Milano, Italy). Fifty images were analyzed for each slide and the tail moment was registered. DNA damage was expressed as the percentage of DNA in the tail [39 (link),40 (link)].

Bacterial strains and phages used in this study are listed in S1 Table. Growth experiments of lactococcal strains and phage propagations were performed as described previously [13 (link), 71 (link)]. TP901-1erm and mutant derivatives were induced from corresponding lysogens of L. lactis NZ9000-Cro_t712 using the following conditions: the relevant strains were grown at 30°C to an A₆₀₀ of 0.3, at which point mitomycin C (Sigma) was added to a final concentration of 3 μg/ml with subsequent incubation at 20°C until lysis occurred. NaCl, 1 M (w/v) final concentration, was added to the resultant phage lysates followed by centrifugation in a Thermo Scientific SL16R centrifuge at 5580 × g for 15 min and stored at 4^°C.

In 2022, 158 various beer samples were acquired from supermarkets and beer shops in 7 different districts of Shanghai. To promote the discussion and comparison of the results, the samples were divided into the following categories: fermentation style—ale (n = 73) and lager (n = 85); alcohol content—nonalcoholic (n = 4), between 3 and 5% vol. (n = 87), and >5% vol. (n = 67); color—pale (n = 23), amber (n = 117), and dark (n = 18); and origin—domestic (n = 121) and foreign (n = 37). Prior to analysis, all beer samples were stored at 4 °C in a dry and ventilated environment. The descriptions of the contaminated samples are organized in Table S1.
Sample pretreatment for each beer was based on a previous study with minor modifications [8 (link)]. An appropriate amount of beer was ultrasonically degassed for 30 min (Geneng G-100S, Shenzhen, China). Then, 10 mL of extraction agent, consisting of ACN:water:AA (70:29:1, v/v/v), was added to 5 ± 0.1 mL of the degassed beer sample. The mixture was vigorously shaken for 30 s and fully ultrasonically extracted for 30 min (Kylin Bell QL 861, Haimen, China). After centrifugation at 5000 rpm for 10 min (Thermo Fisher SL 16R, Waltham, MA, USA), the supernate was filtered through a 0.22 μm organic filter membrane prior to injecting into the UHPLC system. The determination was made using external standard approach.

Ewes were milked twice per day at 07:00 and 17:00 by a milking machine. Individual milk samples (n = 240) were collected at 0, 15, 30, 45, and 60 experimental days for milk chemical composition analysis and at 15, 30, 45, and 60 experimental days (n = 192) for milk FA profile determination and antioxidative status (five aliquots of 15 mL each). The stored milk aliquots were obtained after mixing a yield from the evening and the following morning milk on an identical percent volume (5%). Individual blood samples (n = 192) were also collected at 15, 30, 45, and 60 experimental days from the jugular vein of each ewe after the milking, prior to access on feeds. Approximately 10 mL of whole blood were immediately transferred to heparin-containing tubes (170 units heparin; BD Vacutainer, Plymouth, UK). Then, the blood samples were centrifuged (SL16R, Thermo Fisher Scientific, Waltham, MA, USA) at 2500 rpm for 15 min at 4 °C to separate plasma from the cells. Both the milk and blood samples were stored at −80 °C, before analysis. Furthermore, feed samples from alfalfa hay, wheat straw, and concentrate were collected at the beginning of the experiment, and every time a new concentrate was formulated. Samples of Camelina seeds were also collected for chemical analysis every time a new concentrate was formulated.