Ralstonia solanacearum (R. solanacearum), the causative agent of brown rot disease in potatoes, using efficient and economical medications such as CuO and MgO metal oxide nanoparticles, both in vitro and in vivo, to reduce the risk of pesticide residues.Potatoes are a crucial vegetable crop in Egypt in terms of production and consumption. However, the potato industry suffers significant annual losses due to brown rot disease. This study aimed to suppress R. solanacearum was strongly inhibited by CuO and MgO NPs at a concentration of 3 mg/mL, resulting in zones of inhibition (ZOI) of 19.3 mm and 17 mm, respectively. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CuO-NPs and MgO-NPs were 0.5, 0.6, and 0.6, 0.75 mg/mL, respectively. When applied in vivo through seed dressing and tuber soaking at their respective MIC concentrations, CuO-NPs and MgO-NPs significantly reduced the incidence of brown rot disease to 71.2% and 69.4%, respectively, compared to 43.0% and 39.5% in bulk CuSO4 and bulk MgSO4 treatments, respectively. Furthermore, CuO-NPs and MgO-NPs significantly increased the yield, total chlorophyll content, and enzyme efficiency of potato plants compared with the infected control plants. TEM revealed that the bacterial cytomembrane was severely damaged by nanomechanical forces after interaction with CuO-NPs and MgO-NPs, as evidenced by lipid peroxidation and ultrastructural investigations.CuO and MgO metal oxide nanoparticles were synthesized via a simple chemical process. The average particle size, morphology, and structure of the nanoparticles were characterized using UV-visible spectroscopy, transmission electron microscopy (TEM), zeta potential analysis, X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. The growth of R. solanacearum in potato plants treated with CuO-NPs and MgO-NPs compared to infected control plants highlights the potential benefits for the potato industry in Egypt. Further investigations are needed to explore using metal oxide nanoparticles for treating other plant diseases.The results of this study suggest that CuO-NPs and MgO-NPs can be used as intelligent agents to manage plant pathogens in agriculture. The use of metal oxide nanoparticles could provide a risk-free alternative for treating plant diseases, which are currently one of the biggest challenges faced by the potato industry in Egypt. The significant increase in yield, photosynthetic pigments, enzymatic activity, and total phenol-promoted resistance to  Solanum tuberosum), a crucial vegetable crop in Egypt, is one of the most important crops worldwide. It ranks fourth among the most significant staple foods consumed globally, following wheat, rice, and maize. Egypt produces over 6.4 million tons of potatoes annually from 209 thousand hectares, with an average yield of approximately 30.6 tons per hectare and a market value of approximately $307 million , phosphorus (P), and potassium (K) , sodium hydroxide, and magnesium sulfate, were purchased from Sigma-Aldrich . Louis, MO, USA). i et al. .4 and MgSO4 were prepared and used in this study. To prepare antibacterial stocks, 10 mg of each tested agent was dissolved in a suitable solvent. A suspension of each concentration was applied to R. solanacearum. The plates were divided into four replicates and treated with gentamicin or sterile water as a control. All plates were maintained at a temperature of 28 ± 2 °C and relative humidity of 80 ± 5%. After two days, the bacterial count was recorded using the method described by Tiwari et al.  of CuSOi et al. .In addition, with some modifications, 2.9 g of copper nitrate and 1.2 g of polyvinylpyrrolidone (PVP) were dissolved in 100 mL of deionized water using a magnetic stirrer for approximately two hours. The solution was then heated to 60 °C. Then, 90 mL of 1 M sodium hydroxide (NaOH) was added dropwise to the aqueous copper nitrate solution with vigorous stirring for an hour and a half until a black-brown precipitate formed. The precipitate was centrifuged at 10,000 rpm for 30 min using a Rotina 380 R centrifuge  and placed in an oven at 50 °C for 2 h  and magnesium sulfate were prepared in 100 mL of distilled water. The two solutions were then added dropwise to the solution at a rate of 1 mL/min while stirring at high speed for 1 h. The resulting solid solution was then thoroughly cleaned using deionized water to remove any impurities and dried in an oven at approximately 100 °C for over 24 h. Finally, the product was annealed for 3 h at 450 °C to yield white MgO nanoparticles  of the Agricultural Research Center in Egypt. UV-vis scanning was performed using a Jasco V630 instrument produced in Europe, and the analysis range was set between 200 and 800 nm.The morphology, size, and form of the nanoparticles were analyzed using TEM at the National Research Center in Giza, Egypt. TEM analysis was conducted using two instruments: a Philips CM 12 TEM operated at 120 KV with resolutions up to 510,000 magnifications and a Joel 1230 TEM operated at 100 KV coupled to a CD camera . SAED and HRTEM imaging were performed using a JEOL 3000 F instrument operated at 300 KV with a spot size of 1 nm and an acquisition time of one minute, respectively.The Nano ZS from Malvern, UK, was utilized in all analyses, which were performed at the Central Laboratory for Nanotechnology and Advanced Materials (NAMCL), Agricultural Research Center (ARC), Giza, Egypt. The laser particle analyzer (LPA) was carried out using the Malvern Dispersion Software, with a range of zeta potential (mV) from − 200 to 200 mV and a size range (nm) from 0.6 to 6000, as reported by Li et al. .Powder elemental analysis was conducted using a Rigaku Smart Lab Beijing Co. XRD model . The measurements were performed with a wavelength of 1.54 Å, 45 kV, and 40 mA under a temperature range between 4 and 80° in the angular range of 2θ. The interplanar spacing (‘d’) was calculated using Bragg’s law, 2d sinθ =  λ, where ‘θ’ is the diffraction angle and ‘λ’ is the wavelength of the X-ray used.-1.In addition, FTIR analysis was performed using a Nicolet Avatar 660 FTIR analyzer  at the National Petroleum Research Institute in Egypt, covering a range of 500–4000 cmR. solanacearum were prepared using 24-h-old bacterial cells containing 10–7 colony-forming units per mL (CFU/mL). Sterilized YPG agar medium was thoroughly mixed with 100 µL of the cell suspension and evenly distributed across sterilized Petri dishes. Aqueous nanoparticles (3 mg/mL) with well diameters of 6.0 mm were tested in 100 µL and added to the medium after solidification. The plates were incubated at 30 °C for 48 h, followed by a 2-h pre-incubation at 4 °C. The inhibition zones were measured in millimeters after incubation. Each test was repeated five times for each nanoparticle, along with a control group (gentamicin). Acute toxicity was evaluated after two days to identify inhibition. Before treatment, all the solutions were sonicated for 5 min to reduce sedimentation, particularly at higher doses.This study employed the agar diffusion technique to examine the antimicrobial activity of the nanoparticles. The antimicrobial activity was indicated by the zone of inhibition, as defined by Bonev et al. . Cell suThe measurements were performed by repeated half-fold dilutions of NPs from a diluted stock solution in Mueller-Hinton broth medium, following the method outlined by Azam et al. , with slThe procedure outlined by Roy Choudhury et al.  was follTEM was performed at the Central Laboratory of the Electron Microscope, Fungi Center, El-Azhar University, Egypt.The compost product used in this study, produced by the Ministry of Agriculture in Giza, Egypt, contained compound percentages of organic matter (38.6%), organic carbon (22.4%), C/N ratio (10.5%), total phosphorus (0.26%), and potassium (1.1%). These compounds exert beneficial effects on plant growth. Before application, the compost tea preparation was examined for microorganisms on the PDA medium. Compost tea and humic acid (15.9%) were added at 250 mL/pot before planting. After the soil was infested with the pathogen, pots were sown with disease-free tubers of the Spunta variety. Each treatment was replicated five times, and uninoculated controls were prepared similarly.R. solanacearum in the soil. The density of R. solanacearum was recorded for each treatment. The soil samples were transferred to the laboratory for bacterial counts in potato pots using the SMSA medium  collected from potato districts in El-Sharkia Governorate, Egypt. Each pot contained 6 kg of soil, and one uniform disease-free tuber of the Spunta variety was planted in each pot. The soil infestation was performed using the soil drenching method of Deberdt et al.  at a ratr et al. , with soR. solanacearum isolates were propagated in liquid YPG broth and incubated for four days at 28 °C on a rotary shaker  at 150 rpm. Subsequently, the cultures were centrifuged at 10 °C for 10 min at 10,000 rpm using a centrifuge . The resulting bacterial pellets were optically calibrated to 108 CFU/mL and suspended in distilled water. Measurements were taken using a spectrophotometer  at an optical density (OD) of 600.R. solanacearum. One day after infection, nanoparticles were added again to the infected pots at 180 mL/pot, equivalent to 30 mL/kg soil . The application rate was 1 g/kg of seed.First, a group of potato tubers was soaked in a nanoparticle solution for 30 min before planting. After 21 days of planting, the pots were infected with R. solanacearum were included in the experiment. Throughout the growing season, all pots were placed in a greenhouse with a temperature range of 26–30 °C and a relative humidity of 75–90%. After 85 days of potato planting, various parameters were observed, including the reduction percentage, wilt incidence, and yield per plant. Data on disease severity as a percentage, infection rate, tuber weight (in grams), phenol levels, enzyme activity (polyphenol oxidase and peroxidase), and pigments  were collected.Uninfected controls  and controls infested with Photosynthetic pigments  were extracted from fresh leaf samples in the abovementioned experiments. The extraction was performed using pure acetone based on Fadeel’s method Fadeel . SpectroPeroxidase (EC 1.11.1.7) activity was determined and expressed in milligrams per gram of fresh weight (mg/g fwt). The activity was measured by recording the change in absorbance per minute immediately after adding the substrate, following the method described by Kochba et al. . Color dPolyphenol oxidase (EC 1.14.18.1) activity was measured using the method outlined by Lisker et al. . Color dThe color optical density of the reaction mixture was measured at a wavelength of 520 nm using a spectrophotometer , following the method described by Mechikova et al. .The obtained data were analyzed using ANOVA, and the least significant difference (LSD) test was used to determine statistically significant differences between treatments at a significance level of p < 0.05. Statistical analysis was performed using the CoStat software . The results were presented as mean ± standard error with a sample size (n) of 3, following the guidelines described by Snedecor and Cochran .4 and MgSO4, on the growth of R. solanacearum. As shown in (Table ), bacterial growth decreased with increasing metal concentrations. In contrast, low metal concentrations resulted in the narrowest growth density compared to that of the control.This experiment aimed to investigate the effects of two metals, CuSOin Table , bacteriUltraviolet (UV) spectroscopy revealed the reduction of CuO to CuO nanoparticles, which was confirmed by the peak obtained at 220 nm with an absorption value of 0.2 Fig. .The synthesized CuO-NPs had an average size of 3.59 and 6.05 nm. They were smooth, spherical, loose, and evenly distributed throughout the matrix Fig. .The zeta potential of the CuO-NPs was measured to be − 39.5 mV, indicating the surface charge of the particles. This value suggested that CuO-NPs were stable, as shown in .The X-ray diffraction patterns of the synthesized CuO-NPs were analyzed to determine their crystalline nature. Distinct diffraction peaks were observed at specific 2θ values. Reference code 00-048-1548 was used to identify the crystal structure. The diffraction peaks observed at 35.417°, 35.544°, 38.709°, 38.902°, 48.718°, and 61.526° correspond to the crystal planes (h k l) of (0 0 2), (1 1 –1), (1 1 1), (2 0 0), and (2 0 –2) crystal planes, respectively. Two sharp peaks were particularly prominent at 35.544° and 38.709°, as depicted in  spectroscopy revealed MgO-NPs, which could be confirmed by the narrow peak obtained at 200 nm with an absorption value of 2.5 Fig. .The synthesized MgO-NPs had average sizes ranging from 3.71 to 6.58 nm with spherical shapes .MgO is a single-phase material with a crystalline structure, as confirmed by the X-ray diffraction pattern of MgO-NPs using the reference code (04-007-3846). The diffraction pattern showed several well-defined peaks at 2θ values of 37.016°, 43.006°, 62.447°, 74.869°, and 78.822° correspond to the crystal planes (h k l) of (1 1 1), (2 0 0), (2 2 0), (3 1 1), and (2 2 2), respectively. The two sharp peaks observed were indexed to the (2 2 0) and (2 0 0) planes, as illustrated in Fig. .) and  demonstrate that CuO-NPs exhibited the most effective inhibition of R. solanacearum growth, resulting in the widest zone of inhibition with an average diameter of 19.3 mm. Following CuO-NPs, MgO-NPs demonstrated significant inhibitory activity with an average zone of inhibition diameter of 17.0 mm. Notably, these nanoparticles suppressed pathogen activity considerably better than the control treatment with gentamicin.The diffusion agar technique was employed for the experimental procedure, with a well diameter of 6.0 mm and a volume of 100 µL. The antimicrobial activity of the NPs was directly proportional to the equivalent concentrations of the prepared nanosolutions. The results in Table  and  and illustrated in  demonstrate the effectiveness of metal oxide NPs, specifically CuO-NPs, and MgO-NPs, in reducing disease incidence compared to the control treatment. CuO-NPs and MgO-NPs demonstrated disease reductions of 71.2% and 69.4%, respectively. In contrast, the application of magnesium sulfate at a concentration of 3 mg/mL resulted in the lowest disease reduction (39.5%) compared to the control treatment.The data presented in Table  and illu in Fig.  demonstr4 and MgSO4 were added to the soil, respectively. The values of chlorophyll (b) were 0.221 and 0.351 in the CuSO4 and MgSO4 treatments, respectively. Among the treatments, the lowest carotenoid values were recorded at 0.120 in the presence of CuSO4. In contrast, when CuO-NPs and MgO-NPs were introduced into the soil, the highest values of chlorophyll (a) were 0.769 and 0.642, respectively, whereas the chlorophyll (b) values reached 0.621 and 0.551, respectively. These values were notably higher than those observed for the control treatments (Table ).The observed chlorophyll (a) values were 0.369 and 0.442 when CuSOts Table .4 and CuSO4 were added . These values were significantly lower than that of the Rhizo-N treatment (0.401).Table 4 and MgSO4 were added . These values were notably lower than that of the the Rhizo-N treatment (0.274).Furthermore, the maximum polyphenol oxidase activity values were observed by adding CuO-NPs and MgO-NPs, measuring 0.405 and 0.427, respectively. These values were also within the range typically observed in healthy plants. In contrast, the lowest polyphenol oxidase activity values were recorded when bulk CuSO4 and CuSO4 were added . These values were notably lower than that of the Rhizo-N treatment (2.208).The phenol contents were examined and are presented in Table R. solanacearum, as identified by Yabuuchi et al.  in bacterial growth, as determined by the number of colony-forming units (CFU), with an increase in metal concentration. In contrast, lower metal concentrations did not cause any reduction in CFU numbers compared with the control treatment. These findings are consistent with those reported by Cai et al. , who obsThe application of ultraviolet (UV) visible spectroscopy in this study revealed that CuO-NPs exhibited a surface plasmon absorption band with a peak at 220 nm and an absorption value of approximately 0.2. These results align closely with the findings of Fadhil et al. , who rep− 1, suggesting the presence of different chemical compounds surrounding the metal nanoparticles. These findings align well with those reported by Pandiyarajan et al. , (1 1 –1), (1 1 1), (2 0 0), and (2 0 –2). These results closely correspond to the findings reported by Mobarak et al. , providi− 1. The peak at 669.05 cm− 1 represents a complex fingerprint region encompassing multiple distinct vibrations. The FTIR spectrum of the magnesium nanoparticles indicated the presence of various chemical compounds surrounding them. This finding aligns with a previous conclusion drawn by Imani and Safaei (Ultraviolet-visible (UV-vis) spectroscopy of the MgO-NPs revealed a surface plasmon resonance (SPR) absorption band within the 200–300 nm peak range, with an absorption value of approximately 0.6. Essien et al.  identifid Safaei  that sevThe diffraction pattern obtained from the X-ray analysis of the prepared MgO-NPs in the present study demonstrated that MgO had a single-phase crystalline structure indexed to the (1 1 1), (2 0 0), (2 2 0), (3 1 1), and (2 2 2). The two sharp peaks observed were indexed to the (2 2 0) and (2 0 0) diffraction planes. Previous studies conducted by Imani and Safaei  and MajiShigella flexneri and B. subtilis, depicted high sensitivity indicated by the inhibitory zones created by CuO-NPs. Similarly, Cai et al. (S. aureus) and gram-negative (E. coli) bacteria.The antibacterial activities of the CuO-NPs and MgO-NPs synthesized in this study were investigated. The results demonstrated that CuO-NPs exhibited the highest antibacterial activity, with an inhibitory zone of 19.3 mm on TZC media, surpassing gentamicin. Pandiyarajan et al.  reportedi et al.  found thClavibacter michiganensis subsp. sepedonicus, R. solanacearum, and Dickeya solani, respectively. The present data indicate that the prepared nanoparticles exhibited stronger antibacterial activity than bulk materials at the same concentration, particularly against R. solanacearum. These findings are consistent with the observations of Mondal and Mani , which can damage various biological components, including proteins, lipids, and nucleic acids .The greenhouse experiment involved the application of NPs, which resulted in a significant reduction of bacterial wilt. Specifically, CuO-NPs and MgO-NPs reduced bacterial wilt by 71.2% and 69.4%, respectively. In the absence of nanoparticle treatment, potato tubers exhibited nearly complete mortality. These findings are consistent with those reported by Chen et al. , who obsPhomopsis vexans, R. solanacearum, and Meloidogyne incognita. In addition, researchers have employed seed and soil inoculation methods. The application of 200 ppm ZnO-NPs by spraying resulted in a significant decrease in the disease index, reducing it to one.Khan and Siddiqui  investigConsequently, the utilization of various NPs, as demonstrated in the aforementioned studies, has shown promising potential for increasing crop yields compared to lower concentrations of equivalent bulk materials.In this study, applying CuO-NPs and MgO-NPs to the soil resulted in the highest recorded values of chlorophyll (a) and chlorophyll (b), respectively, compared with the lower values observed with equivalent bulk materials. The use of NPs significantly increased the chlorophyll content, indicating improved plant photosynthetic activity. Previous studies have indicated that nanoparticles can enhance root pore formation, nutrient uptake, and hydromineral flow (Castiglione et al. 4 and MgSO4 bulk materials resulted in the lowest activity values for these enzymes.Moreover, the present data revealed that the application of NPs led to increased peroxidase and polyphenol oxidase activities, as well as enhanced phenolic content. The highest peroxidase and polyphenol oxidase activities were recorded when CuO-NPs and MgO-NPs were used, respectively, and these values were comparable to those observed in healthy plants. Conversely, adding CuSOR. solanacearum, a phytopathogenic bacterium that causes destructive bacterial wilt. A straightforward chemical process was employed to synthesize CuO-NPs and MgO-NPs. The NPs produced were characterized using UV and zeta potential measurements, TEM, and FTIR spectroscopy. Additionally, a parallel experiment was conducted to examine the impact of copper and magnesium sulfate solutions. The experimental data revealed that both CuO-NPs and MgO-NPs exhibited significantly stronger antibacterial activity, with CuO-NPs demonstrating greater biocompatibility. CuO-NPs exhibited lower MIC and MBC values than MgO-NPs.This study was designed to reduce the reliance on pesticides. It focused on comparing and discussing the antibacterial effects of CuO and MgO NPs against Furthermore, TEM imaging revealed that the primary toxicity mechanism involved a rupture of the outer cell membrane, resulting in cellular protein damage. In vivo, the application of CuO-NPs and MgO-NPs influenced the growth of potato plants by reducing the disease index. CuO-NPs and MgO-NPs exhibited control efficacy of 71.2% and 69.4%, respectively. Given their low dosage requirements and potent antibacterial activity, both CuO-NPs and MgO-NPs hold promise as potential biocides for effective plant disease control and offer alternative agricultural disease management strategies. Future research should focus on understanding nanoparticle absorption and transport within the rhizosphere.


Colored potatoes comprise many bioactive compounds that potentially support human health. Polyphenols present in them have associated therapeutic benefits like antimutagenic and anticarcinogenic properties.PP-1901 and Lady Rosetta (LR) potato varieties. Changes in the antioxidant activity, color, total ascorbic acid, phenolic, and flavonoid content were based on the variations in parameters including temperature (blanching using hot water and steam) and capacity 100– 900 W (blanching using microwave).The current study aimed to explore the effects of different blanching methods  on the phytochemical and structural aspects of PP-1901 and LR varieties, all the blanching methods led to a significant reduction in residual peroxidase activity, as well as affecting their color. The preservation of bioactive substances exhibited a microwave steam>hot water blanching trend. Blanching significantly increased the antioxidant activity of all the samples. Additionally, Fourier-transform infrared spectroscopy revealed that phytocompounds were retained to their maximum in microwave-blanched samples, especially at 300 W. The type of blanching method significantly affected the thermal properties of potatoes by disrupting the ordered structure of the matrix.For both Microwaves at 300 W can be used as a novel and suitable alternative technique for blanching potatoes, which successfully retained the original quality of it in comparison to steam and hot water blanching. They also contain niacin, folate, and riboflavin, as well as various phenolic compounds which increase the antioxidant capacity of potatoes (−1) than hot-water treatment (41.41 kJ mol−1) to reach the transition state and then to unfold the polyphenol oxidase enzyme  (Dry matter-24.00 ± 0.12%/Starch-16.23 ± 0.16% dry wt. basis) were procured from the Department of Vegetable Science, Punjab Agricultural University, India. Potato tubers were sliced using stainless steel knife (thicknesses of slice6 ± 0.27 mm and diameter of 25 ± 0.52 mm). A vernier caliper was used to ascertain that the thickness of the slices remained within this range. After slicing they were blanched immediately to avoid enzymatic browning.Potato varieties (purple potato variety) (Dry matter-18.30 ± 0.06%/Starch-13.20 ± 0.12%dry wt. basis) and 2.2.100, MB300, and MB900, respectively. Blanched samples were immediately dipped in water (20°C) to avoid over-cooking. Unblanched potato slices were taken as a control for both varieties, i.e., PP-1901 and LR.Potato slices were subjected to WB (85°C for 5 min) as per the method adopted by Zhang et al.  and were2.3.2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′- azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), 6-hydroxy-2,5,7,8-tetramethyl chroman-2-carboxylic acid (Trolox), Folin–Ciocalteu reagent, gallic acid, sodium carbonate, aluminum chloride, potassium acetate, quercetin, acetic acid, hexane, metaphosphoric acid, and 2,6-dichlorophenol indophenol sodium salt  were procured from Sisco Research Laboratories Pvt. Ltd. (SRL), Mumbai, India. Throughout the experiment, deionized water that had been filtered through a 0.22 μm filter was used.2.4.2.4.1.For estimating moisture content, the standard hot air oven  method was used. Samples were placed inside the oven at 105°C for 24 h . EquatioColor parameters of potato samples were assessed using a handheld hunter lab colorimeter  as per the method described by Aggarwal et al. . The samL*, a*, and b* represents lightness-darkness, redness-greenness, and yellowness-blueness.2.4.2.2O2 (0.3%) and 0.5 ml of guaiacol solution (1%in absolute ethanol v/v). The solution was observed for any color development. The appearance of red color confirmed the presence of POD enzyme activity. For quantitative estimation, 5 g of potato slices were crushed in phosphate buffer (pH 7.8) containing 1% (w/v) polyvinyl/pyrrolidone. The crushed sample was cold centrifuged  at 4°C for 3,000 × g for 10 min. The supernatant (crude enzyme extract) obtained was used for the POD assay. In a final volume of 3.0 ml, the test mixture contained 50 mmol L−1 sodium phosphate buffer (pH 7.0), 12 mmol L−1 H2O2, 7 mmol L−1 guaiacol, and 0.1 ml of enzyme extract. UV Visible spectrophotometer  was used to record the increase in absorbance at 470 nm for 3 min. A change of absorbance from the initial reading to the final reading was taken as residual POD activity  tests were performed for all the samples, as per the method adopted by Liburdi et al. , with slactivity .2.5.2.5.1.The extraction method was adopted from the study by Wu et al. , with so2.5.2.Total phenolic content (TPC) and total flavonoid content (TFC) were estimated as per the methods given by Fattahi et al. , with sl2.5.3.The titrimetric method was used to measure ascorbic acid content using 2,6-dichlorophenol indophenol dye (0.04% w/v) and results were expressed in mg/100 g . Anthocy520nm–A700nm) (pH 1.0)–(A520nm–A700nm) (pH 4.5), DF is dilution factor, MW is the molecular weight of cyanidin-3-glucoside (449.2), W is the weight of the sample (g), and Ɛ is molar absorptivity .Where A is absorbance (A2.5.4.4/100 g DW) were estimated as per the methods explained by Huang et al. , ABTS radical scavenging activity (μg TE/g DW), and Ferric reducing antioxidant powers (FRAP)  . Directl2.6.2.300 at magnifications ranging from ×100 to ×5,000. The sample was mounted on an aluminum stub using double-backed cellophane tape after being coated with gold–palladium (60:40 w/w) in an auto fine coater, JEOL JFC-1600.A scanning electron microscope  was used to morphologically analyze the control and MB2.7.A texture analyzer  outfitted with a 50 kg load cell was used to determine the texture of fresh and treated potato cuboids. Samples were compressed to 50% (3 mm compression) of their original height (6 mm thickness), with a 5 s interval between compressions. The properties of hardness, springiness, cohesiveness, fracturability, and chewiness were measured for samples .2.8.p), onset temperature (To), and conclusion temperature (Tc) were calculated using a thermogram.The differences in the thermal characteristics of the potato samples after each blanching treatment were examined using a differential scanning calorimeter (DSC) as mentioned in a study conducted by Zhang et al. , with sl2.9.p < 0.05% significance level). To determine mean differences, the Tukey post hoc test was used. Points of FTIR were plotted using Origin Pro 2021 .Experiments were replicated thrice for all the samples and the values were reported as mean ± standard deviation. Obtained results were statistically analyzed using Minitab 18.1 software and were subjected to two-way ANOVA (3.3.1.PP-1901 is a purple-colored potato variety containing anthocyanins (900 (67.01) as compared to the control (36.01), denoting a higher amount of anthocyanin disruption which is also confirmed in further sections. These results are consistent with past studies, which showed that the entire color change tended to increase as microwave power levels increased as natural pigments are unstable and sensitive to several elements, including light, temperature, oxidation, pH, metal ions, etc.  in residual POD activity was observed in all blanched samples with MB900 showing maximum reduction, indicating the intensity of enzymatic denaturation brought by high-power microwaves  which can be attributed to the lower amount of thermal shock experienced by POD at a lower microwave power. These findings are in agreement with those observed by Wang et al.  for purple and LR variety, respectively , which w300 W to MB900 W, which can have a similar explanation to that of TPC. In the case of the purple variety, a decrease in TFC was observed. The difference in the results among varieties may be due to the more heat-labile flavonoids (anthocyanins) present in the purple variety  lowered the ascorbic acid content (AAC) for treated potato samples. Out of all samples, WB potatoes had the least retention of AAC in comparison to control samples and had a reduced rate of 52.99 and 36.92% for purple and LR varieties, respectively. Leaching of soluble particles into the blanching water, which has been viewed as the primary drawback of WB, could be one possible explanation for this loss  while this phenomenon is not that effective at lower power levels, whereas higher powers are destructive due to high heat generation  indicate the presence of amide I, 1480–1,575 cm−1 (N-H in-plane bending vibration) amide II, and 1,200–1,400 cm−1 (N-H deformation and C-N stretching) amide III region. Bands with wavenumber 3,200–3,400 cm−1 and 2,800–3,000 cm−1 represent -OH and -CH stretching vibrations, respectively. Absorption around 2,921 cm−1 depicts CH3 asymmetric stretching vibrations in aliphatic chains of protein . The O-H bonds , which are frequently present in both native and hydrolyzed potato starch, can be seen in peaks between 3,100 and 3,700 cm−1. The starch molecules’ CH2 bonds (methylene linkages), which are missing from native potato starch but present in hydrolyzed potato starch, are represented by the wavenumber between 2,920 and 2,928 cm−1  on the textural properties of control and blanched samples, as shown in 900 samples had the lowest value of textural parameters, indicating larger tissue damage to potatoes. A similar kind of reduction in values of hardness, chewiness, and fracturability of radio frequency blanched potato cuboids was observed by Zhang et al.  and enthalpy (∆H) of control and blanched samples of both varieties are shown in g and ∆H of control for LR and PP-1901 samples were found to be 60 and 63°C and 1.69 and 2.73 J/g, respectively. A similar Tg value (60°C) for raw potatoes has been reported by Tian et al. (p < 0.05) both Tg and ∆H. Disruption in the ordered structure of potato tissue which has been confirmed in previous sections might have resulted in decreasing both Tg and ∆H values of potato samples. A decrease in the value of Tg indicates a loss in molecular order and crystalline structure  was related to increased heat generation, which impacted heat-labile phytochemicals found in potatoes. On the other hand, a lower power level (100 W) was ineffective to remove residual POD levels. As a result, the databases employed in the current study provided a broad overview of the blanching methods. An appropriate approach should have a high capacity for inactivating the browning enzyme while retaining the material’s original color and nutritional value. These process parameters could also be used to facilitate the production of other related products like French fries and other processed commodities, which has a huge commercial potential.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.RS: conceptualization, data curation, writing–original draft, preparation, methodology, visualization, and writing–review and editing. SK: supervision, conceptualization, writing–original draft and preparation, methodology, and writing–review and editing. PA: editing and revising. AD: supervision, conceptualization, software, writing–original draft and preparation, methodology, and writing–review and editing.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


To investigate the critical thinking disposition of medical undergraduates.This cross-sectional study was performed on 426 students from four majors, including preventive medicine, maternal and children’s health care medicine, health inspection and quarantine, and food quality and safety. The survey was completed in May 2019 using the California Critical Thinking Dispositions Inventory-Chinese version (CTDI-CV).A total of 435 questionnaires were distributed and 426 valid questionnaires were collected, with an effective rate of 97.93%. The CTDI-CV overall average score was 262.02 ± 34.74 points indicating an ambivalent disposition in medical undergraduate students. Only one of the subscales (maturity in judgment) had mean scores of 43.35 ± 8.23 indicating the positive disposition of students. Among them, males scored 257.42 ± 35.06 lower than females’ 264.82 ± 34.32, the difference was statistically significant. The target scores of preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety were 265.17 ± 30.10, 260.26 ± 37.05, 271.73 ± 33.55, and 252.11 ± 39.87, respectively. The difference was statistically significant. Among the three dimensions of seeking truth, open mind, and cognitive maturity, the scores of males were 38.26 ± 7.48, 38.78 ± 6.46 and 41.03 ± 8.69, which were lower than females’ 39.97 ± 7.11, 40.48 ± 6.48 and 44.91 ± 7.60, respectively. The scores of food quality and safety students were 37.23 ± 7.08, 36.61 ± 7.41 and 40.57 ± 8.60, respectively, which were lower than the preventive medicine .Most medical students were found to have an ambivalent disposition which meant they were not disposed toward critical thinking. These findings suggested that more effective teaching methods should be taken to facilitate critical thinking disposition and problem-solving ability. Due to the multifaceted and complex decisions of prevention doctors, high-level cognitive abilities and professional techniques are required. There is increasing recognition that prevention doctors facilitate to prevention and control of the disease for humans, including infectious diseases and chronic non-communicable diseases. To achieve these optimal disease prevention and control outcomes, prevention doctors are required to provide evidence-based, effective protective, and population services. Prevention doctors need well-developed cognitive skills which need positive critical thinking abilities to make independent decisions. Despite the importance of critical thinking disposition, there is limited evidence focused on thinking processes in prevention medical students. Hence, we try to investigate the distribution of critical thinking disposition in medical undergraduates in this study.In the education domain, there is disagreement about the term “critical thinking”. Predominant researchers defined “critical thinking” as a rational process involving ‘interpretation, analysis, evaluation, inference, explanation, induction, numeracy, deduce, and self-regulation’ . Other rThe evidence has identified that there is a correlation between leadership and positive disease outcomes, including precise decisions and more health services, lower incidence, prevalence, and mortality of diseases, and higher healthy satisfaction. The level of cognition, leadership, and critical thinking abilities exert crucial roles in judgment and the ability to decision-making. Critical thinking skills can develop the ability for decision-making and problem-solving , which fThe most commonly used measures to evaluate critical thinking abilities are standardized, commercially available tools such as the California Critical Thinking Skills Test (CCTST) , CriticaThe primary aim of this study was to better understand the characteristics that elicit students’ critical thinking abilities in the medical university, which provides a scientific basis for curriculum adjustment and modification. The role of educators is to teach students how to monitor their thinking and make themselves to better resolve problems with more thoughtful thinking. In this current study, we focused on preventive medicine, food quality and safety, and health inspection and quarantine since critical thinking assessments were already available for these disciplines. Specifically, we investigated the features of critical thinking abilities in undergraduate students.While most critical thinking instruments in medical contexts have undergone some form of validation to investigate the features of students in nurse disciplines, to our knowledge none have explored the characteristics of preventive medical students’ critical thinking. This research provides new insight into the characteristics of undergraduate students’ critical thinking abilities, which can further be applied by educators to incorporate more effective critical thinking opportunities and contents in the classroom. These findings suggested the status of critical thinking abilities at the undergraduate level at Medical University, which provides new insight into the way of curriculum modification.A cross-sectional survey of medical undergraduates from Anhui Province university was conducted in May 2019 using a convenience sampling method. The investigation inclusion criteria were: (1) agreed to participate in the investigation and no history of participation in similar studies of critical thinking tests, (2) at least six months of the study period in three subjects of medical university. The investigation exclusion criteria were: unwilling students. The study was approved by Anhui Medical University’s ethical committee (No.20170291). Students were informed of the aim of the research. Informed consent was obtained from all subjects and the questionnaire was completed anonymously in class. A total of 435 students from three academic years enrolled in a survey of critical thinking. Finally, 426 students consented to participate in the research study, which consisted of 265 females and 161 males. After the allocation, nine students were excluded because they could not complete the questionnaire.Data were collected in May 2019 using the California Critical thinking disposition inventory Chinese version (CTDI-CV), which assesses the critical thinking disposition of undergraduate medical students. CTDI-CV, revised from the CCTDI, is a self-reported questionnaire with a six-point Likert-type scale , which cThe investigator approached students in their classrooms and explained the aims and features of the study. Furthermore, the investigator elucidates the feature and instructions of the CTDI-CV on how the test would be taken were explicated to the participants. In addition, participants were requested not to discuss the test with other students.t-test or one-way analysis of variance (ANOVA) with LSD post hoc test or Tamhane’s test. A level of statistical P < 0.05 was set for all the tests and considered to be statistically significant.Statistical analysis was performed for all quantitative data by using SPSS software version 23.0. Data were analyzed using descriptive . Critical thinking abilities evaluation data were analyzed by independent sample Four hundred and thirty-five students completed the critical thinking disposition assessment. Among these, 426 were valid samples. However, 9 were incomplete in the questionnaire. Exclusions were based on the fact of nine students did not fulfill responded to all test items. Table The distribution of critical thinking disposition in medical students is listed in Table The CTDI-CV scores are presented in Table t=-2.139, P = 0.033). In terms of academic levels, the CTDI-CV scores are 263.64 ± 35.47, 260.82 ± 33.06, and 262.37 ± 35.67 in the fifth year, fourth year, and second year, respectively, whilst there is no significant difference among academic levels . In terms of academic majors, the CTDI-CV scores are 265.17 ± 30.10, 260.26 ± 37.05, 271.73 ± 33.55, and 252.11 ± 39.87 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst there is a significant difference among academic levels . After ANOVA with LSD analysis, the results demonstrated that the CTDI-CV scores in students of preventive medicine and health inspection are significantly higher than that in food quality and safety.The CTDI-CV scores in students were 262.02 ± 34.74. Among these, the CTDI-CV target scores in male (257.42 ± 35.06) is significantly lower than that in female (264.82 ± 34.32) . In terms of academic levels, truth-seeking subscale scores are 40.24 ± 7.44, 38.78 ± 7.27, and 39.44 ± 7.29 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, truth-seeking subscale scores are 39.98 ± 7.07, 39.04 ± 7.53, 40.96 ± 6.98, and 37.23 ± 7.08 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst significant difference among groups . After ANOVA with LSD analysis, the results demonstrated that the truth-seeking subscale scores in students of preventive medicine and health inspection are significantly higher than that in food quality and safety.We analyzed the subscale scores in truth-seeking of CTDI-CV. The truth-seeking subscale scores in male (38.26 ± 7.48) is significantly lower than that of female (39.97 ± 7.11) . In terms of academic levels, open-mindedness subscale scores are 39.66 ± 6.75, 39.95 ± 5.81, and 39.82 ± 6.52 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, open-mindedness subscale scores are 40.60 ± 5.96, 39.80 ± 6.59, 41.73 ± 5.71, and 36.61 ± 7.41 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst significant difference among groups . After ANOVA with LSD analysis, the results demonstrated that the open-mindedness subscale scores in students of food quality and safety are significantly lower than the other academic majors.We analyzed the subscale scores in open-mindedness of CTDI-CV. The open-mindedness subscale scores in male (38.78 ± 6.46) is significantly lower than that of female (40.48 ± 6.48) . In terms of academic levels, analyticity subscale scores are 35.68 ± 6.28, 34.33 ± 5.60, and 34.59 ± 5.75 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, analyticity subscale scores are 34.51 ± 5.49, 34.53 ± 5.98, 36.53 ± 6.61, and 34.38 ± 5.52 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst no difference among groups .We analyzed the subscale scores in the analyticity of CTDI-CV. The analyticity subscale scores are no difference between males (34.78 ± 5.83) and females (34.55 ± 5.75) . In terms of systematicity levels, systematicity subscale scores are 36.04 ± 6.28, 35.91 ± 6.34, and 36.85 ± 6.02 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, systematicity subscale scores are 36.89 ± 5.76, 36.04 ± 6.43, 38.27 ± 5.45, and 35.36 ± 6.74 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst no difference among groups .We analyzed the subscale scores in the systematicity of CTDI-CV. The systematicity subscale scores in males (36.09 ± 6.10) and females (36.66 ± 6.20) . In terms of systematicity levels, confidence in reasoning subscale scores are 34.45 ± 6.89, 33.93 ± 7.43, and 33.68 ± 7.35 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, confidence in reasoning subscale scores are 33.84 ± 7.78, 33.40 ± 6.96, 35.93 ± 6.51, and 34.12 ± 7.20 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst no difference among groups .We analyzed the confidence in reasoning subscale scores of CTDI-CV. The confidence in reasoning subscale scores in males (33.73 ± 6.92) and females (33.93 ± 7.55) . In terms of systematicity levels, inquisitiveness subscale scores are 35.06 ± 6.46, 34.40 ± 6.32, and 34.38 ± 6.55 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, inquisitiveness subscale scores are 34.95 ± 6.45, 33.92 ± 6.53, 35.80 ± 6.51, and 33.84 ± 6.19 in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst no difference among groups .We analyzed the subscale scores in the inquisitiveness of CTDI-CV. The inquisitiveness subscale scores in males (34.74 ± 6.29) and females (34.31 ± 6.56) . In terms of systematicity levels, maturity of judgment subscale scores are 42.51 ± 8.32, 43.52 ± 7.75, and 43.62 ± 8.51 in the fifth year, fourth year, and second year, respectively, whilst no difference among groups . In terms of academic majors, scores of maturity of judgment subscale are 44.44 ± 6.97, 43.53 ± 8.93, 42.50 ± 9.41, and 40.57 ± 8.60 in preventive medicine, maternal and child health medicine, health inspection and quarantine, and food quality and safety, respectively, whilst significant difference among groups . After ANOVA with LSD analysis, the results demonstrated that the maturity of judgment subscale scores in students of preventive medicine is significantly higher than that in food quality and safety.Finally, we analyzed the subscale scores in the maturity of judgment of CTDI-CV. The maturity of judgment subscale scores in males (41.03 ± 8.69) is significantly lower than that of females (44.91 ± 7.60) (This study investigates the critical thinking disposition in medical undergraduate students. The results indicate an ambivalent range of critical thinking disposition in medical undergraduate students, including truth-seeking, open-mindedness, and maturity in judgment. However, the present findings resemble those of Zia  and NguyAlternatively, the results further confirmed that critical thinking disposition in male students is lower than that of female students, including truth-seeking, open-mindedness, and maturity of judgment. The critical thinking abilities of students are related to reading and writing but are not easily influenced by educational methods. Some studies such as that of Boso suggests a positive critical thinking disposition with 296 points in nursing students , and ZiaMoreover, in this current study, a high score on the maturity of judgment subscale suggested that students might own the ability to cope with complex situations, which might mean they have the ability of inductive reasoning . InteresThere are some limitations in this current study that should be mentioned. First, a limited sample pool in this study, which is undergraduate students enrolled in preventive medicine, maternal and children’s health medicine, health inspection and quarantine, and food quality and safety, limits the generalization of conclusions. It needs to recruit more subjects representative sample of undergraduate students in future studies. Second, for some reason, we have not included the individual information in this investigation, such as family status, neighbors and partners, urban or rural origin, and family economic levels. Finally, cross-sectional was selected in this study to investigate the nature and characteristics of critical thinking disposition in undergraduate students in medical universities. Further longitudinal studies and intervention field tests should be used to explore the effective method to develop the critical thinking disposition and critical thinking skills in undergraduate students in medical universities.The finding in this present study indicated ambivalent critical thinking disposition in medical undergraduate students. Furthermore, critical thinking disposition in male undergraduates are lower than that of female undergraduates, such as truth-seeking, open-mindedness, and maturity of judgment. Moreover, the academic year might not be the predictor of critical thinking disposition, and this result is inconclusive with the result of other studies , 28, 29.


To investigate the correlation between FF, R2* value of IDEAL-IQ sequence and bone mineral density, and to explore their application value in the osteoporosis.We recruited 105 women and 69 men aged over 30 years who voluntarily underwent DXA and MRI examination of lumbar spine at the same day. Participants were divided into normal, osteopenia and osteoporosis group based on T-score and BMD value of DXA examination. One-way ANOVA was adopted to compare the quantitative parameters among the three groups. Independent samples t-test was utilized to compare FF and R2* value between men and women.Pearson correlation analysis was used to research the correlation between FF, R2* value and BMD.p < 0.05). No significant difference was found in FF value between male and female group, while R2* value were significantly different. Vertebral FF was moderately negatively correlated with aBMD, especially in women . R2* was mildly to moderately positively correlated with aBMD in men , but not in women. Moreover, FF was positively correlated with age, R2* was negatively correlated with age in men, and BMD was negatively correlated with age.Age, height, weight, BMD and FF value were significantly different among three groups (The vertebral FF value of IDEAL-IQ sequence has the potential to be a new biological marker for the assessment of osteoporosis. Vertebral FF is moderately negatively correlated with aBMD, especially in women, allowing accuratly quantify the bone marrow fat. R2* value is mildly to moderately correlated with BMD in men and can be served as a complementary tool in the assessment of osteoporosis. Osteoporosis(OP) is defined as a systemic metabolic bone disease that is characterized by decreased bone mass and microarchitectural deterioration of bone tissue, which leads to increased bone fragility and elevated fracture risk. As of August 2020, the global prevalence of osteoporosis was approximately 18.3%, with the prevalence in women at approximately 23.1% and in men at about 11.7% , osteopenia (− 2.5 SD < T-score <  − 1.0 SD), and osteoporosis (T-score ≤  − 2.5 SD) .A 3.0 T MR scanner  was used. The scanning parameters for the IDEAL-IQ sequence were as follows: TR 6.3 ms, TE 2.9 ms, FOV 38 cm × 38 cm, slice thickness 5 mm, matrix 16 cm × 16 cm, flip angle 4°, NEX 2, number of slices 72, and scan time of the sequence 14 s. After the MRI examination was completed, the L1–L3 vertebral FF and R2* value were measured independently by two radiologists with 2 years of experience using a GE AW 4.6 post-processing workstation. On the FF and R2* map, the sagittal midplane was selected, and the rectangular ROI  was manually outlined to directly obtain the FF and R2* value of each vertebra, and calculated the average FF and R2*. FF maps and R2* maps of three females are shown in Fig. In addition, 30 females and 30 males were randomly selected from all data sets using a random table generated by SPSS to measure FF and R2* value and assess intra-observer reliability. These 60 subjects were also analyzed by another radiologist with 2 years of experience to investigate inter-observer reliability.P-value less than 0.05.Statistical software  was used for data analysis. The data conforming to normal distribution were expressed as mean ± standard deviation (SD). The differences among normal, osteopenia and osteoporosis group in terms of normal variables were compared through One-way ANOVA. Independent samples t-test was utilized to compare FF and R2* value between men and women. Pearson correlation test was applied for analyzing the correlation between two different parametric variables. Reproducibility of the data was assessed by the root mean square coefficient of variation (RMS-CV), and inter- and intra-observer agreement of measurements between two radiologists was assessed by intraclass correlation coefficient (ICC).Differences were considered statistically significant at a The reproducibility of the FF and R2* measurements was satisfactory, with RMS-CV of 2.41% and 2.63%, respectively. The inter- and intra-observer agreements of FF and R2* value were excellent, with ICCs ranging from 0.971 to 0.990 (> 0.9).With DXA as diagnostic criteria, there were significant differences in age, height, weight, T-score, aBMD and FF value among the normal, osteopenia and osteoporosis group, except for BMI and R2* value Table .Table 1P < 0.05)  Table .Table 2p < 0.001. FF was positively correlated with age . However, R2* value exhibited no significant correlation with age. The results are shown in Table Age was negatively correlated with T-score and aBMD value, and the correlation coefficients were -0.498 and -0.494, 1 = -0.527, P1 < 0.001; r2 = -0.515, P2 < 0.001 respectively , but there was no correlation with T-score.Vertebral FF was negatively correlated with T-score and aBMD, with correlation coefficients of approximately: rr = -0.409, P < 0.001), positively correlated with T-score and aBMD value, but no correlation with FF   Table .Table 6, contributing to indirectly estimate bone density and strength [IDEAL-IQ sequence is a new MRI quantitative technique with the parameters FF value, which accurately quantifies bone marrow fat, and R2* value, which reflects iron deposition in vivostrength . Vertebrstrength , 16. TheBone mineral density explains approximately 70% of bone strength, while the remaining 30% is affected by other elements, of which bone marrow adiposity is closely related to bone density and bone strength. Bone marrow adipocytes and osteoblasts are both derived from mesenchymal stem cells (MSCs), and there is a competitive inhibitory relationship between them. With aging, oestrogen deficiency and glucocorticoid exposure, MSCs preferentially differentiate into adipocytes, resulting in reduced bone formation . Some stIron deposition has also been indicated to depress bone density and strength, and raise the incidence of osteoporosis . Iron meSignificant differences were found in age, height and weight among different bone mass groups. Bone loss was more pronounced in subjects who tended to be short and thin, and osteoporosis was more likely to occur. Fahimfar et al.  found thr = -0.494 ~ -0.498, P < 0.001), positively correlated with FF value  but not correlated with R2* value. When subjects were divided by gender, R2* value of males was negatively correlated with age, and mildly to moderately positively correlated with aBMD, while R2* value of females was not significantly correlated with other parameters. The correlation coefficients between FF value and age, aBMD in females were 0.725 and -0.638 respectively, while in males the correlation coefficients were 0.506 and -0.259, which meaned the correlations were more significant in women than in men.No significant difference was found in FF value between male and female, while R2* value was obviously different. There was variability in both FF and R2* value between men aged < 50 years and ≥ 50 years and between menopausal and non-menopausal women, suggesting that FF was primarily age-related, while the difference in R2* may be due to difference in gender. When subjects were not divided by gender, age was negatively correlated with T-score and aBMD value , which was much higher than our results . We speculated that the age of included participants over 50 years and the small sample size may be responsible for this. Chang et al. [At present, the correlation analyses between FF and BMD value are relatively common. Zhao et al.  found thg et al. , adoptinr = -0.219, 0.290, P < 0.05), and the diagnostic efficacy of R2 * in diagnosing osteoporosis was relatively poor. Kim et al. [In recent years, quantitative assessment of FF and R2* value of lumbar spine has been performed, and FF and R2* are considered to be probably moderate markers of osteoporosis , 32. R2*m et al.  selectedThere are several limitations in our study. Firstly, the sample size was relatively small and the number of patients with osteoporosis was relatively insufficient, especially in men. Secondly, MRI examination included L1-L3 vertebrae while DXA measured aBMD of L1-L4 vertebrae, and the difference may influence the findings. What’s more, the incidence of vertebral fractures in our study was particularly low and did not allow for assessment of fragility fracture risk, which has been a hot topic in the research of osteoporosis currently.In short, IDEAL-IQ sequence is a fast, convenient, non-invasive and non-radiation technique. The parameter FF performs a more significant correlation with T-score and aBMD in women than in men, therefore, FF value has the potential to be a new biological marker for the assessment of osteoporosis, predicting the risk of fragility fractures and assessing the efficacy of treatment in women. In contrast, R2* value is mildly to moderately correlated with BMD in men and can be served as a complementary parameter to FF and BMD in the assessment of osteoporosis.


In the 2020–2021 season, C+ had a significantly higher number of MI compared to C− in both championships . Multiple linear regression analysis confirmed that belonging to C+ in the season 2020/2021 was the variable that most strongly influenced the probability of having a muscle injury. Survival analysis revealed a hazard ratio of 3.73 (95% CI 3.018 to 4.628) and of 5.14 (95% CI 3.200 to 8.254) for Serie A and LaLiga respectively. We found an association between SARS-CoV-2 infection and increased risk of muscle injury, emphasizing the importance of carefully considering the infection in the decision-making process for returning to sport. Therefore, SARS-CoV-2 infection should be judged as a real injury requiring specific assessment and training programs.A retrospective cohort study on professional soccer players from the Serie A and LaLiga was conducted to investigate the correlation between SARS-CoV-2 infection and muscle injuries. Players were divided into two groups based on whether they contracted the SARS-CoV-2 infection (C+) or not (C−) during the 2020/2021 season. In the 2019–2020 season, both championships showed a similar number of muscular injuries (MI) between C+ and C− (Serie A:  In late December 2019, The World Health Organization (WHO) office in China was informed about novel cases of pneumonia of unknown aetiology detected in the city of Wuhan, Hubei province [Football (soccer) is a physically demanding sport that requires a high level of neuromuscular readiness, including strength, reactivity, and muscular power ,4. The cHowever, these studies did not show whether having contracted SARS-CoV-2 infection can be considered an additional risk factor for the occurrence of injuries. Recently only the prospective study by Wezenbeek et al., involving three Belgian professional male football teams, during the first half of the 2020–2021 season reported a five times higher risk of developing a muscle strain after a SARS-CoV-2 infection.The goal of this study is to deepen the results reported by Wezenbeek et al. and to retrospectively verify, on a larger sample, the correlation between SARS-CoV-2 infection and muscle injuries among professional footballers of two different championship Serie A and LaLiga. The choice of these two different championships was because Serie A represented the exception of post-lockdown injuries increase and both Serie A and LaLiga had the most active collaboration in identifying and communicating positive cases . Our hypA retrospective cohort study was conducted according to the STROBE guidelines . The aimThe grouping was performed as follows: (1) all football players infected during the 2020–2021 season were classified as “C+”, while all players who did not contract the infection during this time period were classified as “C−”; (2) players who contracted the infection during the 2019–2020 season or during off-season periods  were excluded from the study  and their data were removed from both seasons. This created a database with pre-COVID (2019–2020 season) and post-COVID (2020–2021 season) data.https://www.transfermarkt.co.uk/, accessed on 12 September 2022) a website founded in Germany in May 2000 which contains a wealth of football information from leagues around the world, including data on the type and severity of injuries. This methodology is consistent with previous studies [Data collection was conducted using the Transfermarkt24 site  and 649 players from LaLiga (mean age = 28.2 ± 4.2 years), for a total of 1283 elite football players. Of the Italian championship players, 171 (27.3%) were infected with SARS-CoV-2, with 29 excluded due to being infected during the 2019-2020 season. In LaLiga, a total of 165 (25.4%) players were infected with SARS-CoV-2, with 38 excluded as they were infected during the 2019–2020 season . p = 0.001 95% CI: 117.22 to 451.170) and for matches played (p = 0.015 95% CI: 0.44 to 4.15). In the 2019–2020 Serie A championship, on average each player participated in 21 soccer matches with an average of 1502 ± 898 min played. On average group C+ played for 1719 ± 928 min in 23.1 ± 9.7 matches, while group C− played for 1440 ± 879 min in 20.8 ± 9.9 matches, showing a significant difference between the two groups both for minutes played (p = 0.997 95% CI: −173.67 to 174.38) and for matches played (p = 0.554 95% CI: −2.57 to 1.38). In the 2020/2021 season, on average each player participated in 22 soccer matches with an average of 1453 ± 927 min played, group C+ played on average for 1454 ± 912 min in 21.5 ± 10.1 matches, while group C− played for 1456 ± 934 min in 22.12 ± 10.6 matches, showing a non-significant difference between the two groups both for minutes played  and an average of 265 ± 893 more minutes played in the 2019/2020 season  and an average of 16 ± 883 more minutes played in the 2020/2021 season .The C+ group reported a mean change between seasons of 1.6 ± 9.8 higher number matches played in the 2019/2020 season  were found in the average number of muscular injuries between the C+ group (0.52 ± 0.75) and the C− group (0.43 ± 0.68). In the 2020–2021 season, the C+ group had a significantly higher number of muscular injuries (1.44 ± 0.96) compared to the C− group (0.56 ± 0.78)  ; the C− group reported a mean change between seasons of 0.131 ± 0.843 higher number of muscular injuries in the 2020–2021 season .In the 2019–2020 Serie A championship, non-significant differences (o 1.038) . The C+ The multiple linear regression analysis revealed that, in the 2019–2020 season, muscular injuries were mainly related to age, minutes played, and matches played by the athletes . In the p = 0.401). However, during the 2020–2021 season, the difference in injury rates between the two groups became statistically significant (p < 0.05), with a hazard ratio of 3.73 (95% CI 3.018 to 4.628).The survival analysis based on the Kaplan–Meier curves  revealedp = 0.673 95% CI: −149.36 to 231.33) and for matches played (p = 0.688 95% CI: −1.69 to 2.57). In the 2019–2020 LaLiga championship, on average each player participated in 22 soccer matches with an average of 1559 ± 964 min played. On average C+ group played for 1592 ± 929 min in 22.6 ± 11 matches, while group C− played for 1550 ± 974 min in 22.2 ± 10.7 matches, showing a non-significant difference between the two groups both for minutes played (p = 0.202 95% CI: −299.96 to 63.69) and for matches played (p = 0.129 95% CI: −3.57 to 0.45). In the 2020/2021 season, on average each player participated in 23 soccer matches with an average of 1500 ± 921 min played. Group C+ played on average for 1406 ± 873 min in 21.6 ± 9.9 matches, while group C− played for 1525 ± 933 min in 23.1 ± 10.2 matches, showing a non-significant difference between the two groups both for minutes played (p = 0.273 95% CI: −0.874 to 3.066) and on average 184 ± 969 more minutes played in the 2019/2020 season  and on average 39 ± 1017 more minutes played in the 2019/2020 season .The C+ group reported a mean change between seasons of 1.1 ± 11.1 higher number of matches played in the 2019/2020 season . However, during the 2020-2021 season, the average number of injuries was significantly higher  in the C+ group (1.23 ± 1.04) compared to the C− group (0.32 ± 0.59) ; the C− group reported a mean change between seasons of 0.131 ± 0.843 lower number of muscular injuries in the 2020–2021 season .In LaLiga, the results indicated that during the 2019-2020 season, there was no significant difference in the average number of muscular injuries between the C+ and C− groups, with 0.40 ± 0.67 and 0.39 ± 0.65 injuries, respectively ( ± 0.59) . The C+ p < 0.05), while games played and minutes played by athletes did not significantly influence the occurrence of injuries. However, in the 2020–2021 season, the group variable had a significant influence on the muscle injury rate , while in the season 2020–2021, the difference becomes statistically significant (p < 0.05), with a hazard ratio of 5.14 (95% CI: 3.200 to 8.254).Moreover, in the Spanish championship, the survival analysis  showed hThe main aim of this study was to determine whether professional soccer players who contracted SARS-CoV-2 had an increased risk of muscular injuries. The findings of this study reported a three to five times increased risk of muscle injury after SARS-CoV-2 infection.Over the years, UEFA has conducted numerous studies on the match/minute ratio played by the elite athletes of the teams participating in its international tournaments to define a “normal” value for the incidence of injuries in football ,23,24. IThe results of minutes and matches played showed that in Serie A the players in the C+ group, during the 2019/2020 season, played more games and in total more minutes than the players in group C−; in the 2020/2021 season, the players in group C+ (which contracted COVID-19 during this season), they played fewer games and in total fewer minutes than in the previous season, on the contrary the players in group C− obtained an increase in total games and minutes played. This could be caused both by the period of absence linked to the infection itself and by the greater number of muscle injuries that players in group C+ have suffered, forcing them to have a longer period of absence from sporting activity. The same, albeit to a lesser extent, is observed in the Spanish league showing a substantial overlap of minutes played and games played in the 19/20 season between the two groups; however, in the following season, there was a reduction in minutes and games played in group C+ and an increase in games played and a slight reduction in minutes played in group C−. Our results indicate that SARS-CoV-2 infection could be a risk factor for muscular injury, in line with the recent study by Wezenbeek et al. . Multipl2 max [As discussed by Wezenbeek et al. , there a2 max  are possThese results emphasize the importance of carefully considering the infection in the decision-making process for determining athletes’ readiness to return to sport (RTS). In fact, as already described by Elliot et al.  in additOne possible limitation of this study is that the muscle injury events were collected from a single database, which may have resulted in some injuries being overlooked, omitted, or interpreted differently, leading to over- or underestimation. Another limitation of this study is that it was not possible to control for some factors that may have influenced the increased risk of injury, such as previous muscle injuries in unexamined seasons, severity of COVID-19-related symptoms, total period of quarantine and injured body part. Nevertheless, the study’s strength lies in its design, with a largely homogeneous population group of male professional soccer players followed prospectively with a standardized methodology.This retrospective cohort study, conducted on a large sample of professional male football players from two main European leagues, has revealed a significant association between SARS-CoV-2 infection and increased risk of muscular injury. The results of this study show that the risk of muscular injury is 3.7 to 5.1 times higher in football players who have been infected with SARS-CoV-2. Overall, our study highlights the need for further investigation into the effects of SARS-CoV-2 on athletes and the development of tailored rehabilitation protocols to ensure a safe return to play.


Milk fat is the most variable nutrient in milk, and recent studies have shown that rumen bacteria are closely related to milk fat. However, there is limited research on the relationship between rumen bacteria and milk fatty. Fatty acids (FAs) are an important component of milk fat and are associated with various potential benefits and risks to human health.In this experiment, forty-five healthy Holstein dairy cows with alike physiological and productive conditions were selected from medium-sized dairy farms and raised under the same feeding and management conditions. The experimental period was two weeks. During the experiment, raw milk and rumen fluid were collected, and milk components were determined. In this study, 8 high milk fat percentage (HF) dairy cows and 8 low milk fat percentage (LF) dairy cows were selected for analysis.Prevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03 in HF group were significantly higher than those in the LF group. Spearman rank correlation analysis indicated that milk fat percentage was positively related to Prevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03. Furthermore, Prevotellaceae_UCG-001 was positively related to C14:0 iso, C15:0 iso, C18:0, Ruminococcus_1 with C18:1 t9, Lachnospiraceae_AC2044_group with C18:1 t9 and C18:1 t11, U29-B03 with C15:0 iso.Results showed that the milk fat percentage in HF group was significantly greater than that of the dairy cows in the LF group. 16S rRNA gene sequencing showed that the rumen bacterial abundance of HF dairy cows was significantly higher than that in LF dairy cows; at the genus level, the bacterial abundances of To sum up, rumen bacteria in dairy cows are related to the variation of milk fat, and some rumen bacteria have potential effects on the deposition of certain fatty acids in raw milk. The content of milk fat is generally 3% ~ 5% in milk, and the main component is triglyceride, which account for about 98% of milk fat. The remaining components of milk fat include diacylglycerol, monoacylglycerol, phospholipid, glycolipid, cholesterol, free fatty acids and fat-soluble vitamins . Fatty aFatty acid is the main component of milk fat, and its type, composition and content have been widely concerned in the current years. The milk fatty acid was mainly synthesised by rumen microbia and exogenous uptake . ButyricDialister, Megasphaera, Lachnospira and Sharpea in rumen .n = 8) and LF group (n = 8) according to the milk fat percentage. The feed intake and milk yield between the two groups were similar, which avoided the large difference in milk fat percentage caused by the huge difference in milk yield, and met the requirements of experiment.This experiment was carried out in Tianjin Mengde Dairy Farm, with more than 1,500 Holstein dairy cows in spring. The dairy farm has advanced facilities and records the milk yield of individual dairy cows every day. From more than 1,500 lactating dairy cows, 45 healthy dairy cows with similar body weight, parity (1.3 ± 0.74), day in milk (153 ± 16d) and daily milk yield (34 ± 6 kg) were selected. After collecting milk samples, MilkoScan FT120  was used to determine the content of milk protein, milk fat, lactose, and total solids. The data were subjected to quality control through the mean and standard deviation (SD), and split into HF group  reagent. The purity of extracted DNA was checked using 1% agarose gel, and the DNA concentration was determined using a NanoDrop 2000 UV–vis spectrophotometer . The purified DNA as template, 341F (5’-CCTACGGGNGGCWGCAG-3′) and 806R (5’-GGACTACHVGGGTWTCTAAT-3′) as primers, the V3 to V4 of bacterial 16S rRNA gene was amplified using an ABI GeneAmp 9,700 PCR thermocycler . The PCR amplification conditions and the specific steps of sequencing all adopt the method of http://drive5.com/uparse/) was used to cluster the operational taxonomic unit (OTU) with 97% similarity cutoff, then identified, and removed the chimeric sequences. The longest read was deemed as a representative sequence of the taxonomy for each OTU. Further, those representative sequences were identified against to the Silva SSU128 database using RDP Classifier.All raw reads analysis and the quality control were referenced to the methods as reported by 2.5.3OH solution (20 g/L) was added, and saponification and alkali-catalyzed methyl esterification at 50°C for 20 min in a water bath. Subsequently, Acid-catalyzed methyl esterification was carried out by adding 2 mL of acetyl chloride-methanol solution (100 mL/L) at 90°C for 120 min in a water bath. After cooling, add ultrapure water, transfer the n-hexane phase, and dilute to 10 mL. Add about 1 g of anhydrous sodium sulfate, vortex for 30 s, and let stand for 10 min. Pipette 200 μL of the n-hexane phase into the bottle and dilute the volume to 1 mL, so that the dilution factor of liquid milk to sample bottle is 25. Analysis with GC–MS.The test was carried out by the method independently developed by our laboratory. Draw 2 mL of liquid milk sample, add 25 μL of internal standard C19:0 fatty acid methyl ester , add 4 mL of n-hexane/isopropanol (V/V = 3/2) mixed solution, after vortexing for 30s, centrifugation  for 3 min. Pipette the n-hexane phase into a high temperature test tube with a lid, add n-hexane, centrifuge, take the upper n-hexane phase, repeat twice. To the n-hexane phase containing the lipid extract, 2 mL of NaOH-CHTesting equipment: Gas chromatography tandem mass spectrometer 7890A/7000B (G-150). The conditions of use of the instrument are as follows, chromatographic column: CP-Sil 88; programmed temperature; gas: helium; constant pressure: 38 psi; inlet temperature: 250°C; quadrupole temperature: 150°C; injection volume: 1 μL.2.6.3-N and VFAs, and the relationship between rumen bacteria and raw milk fatty acid composition were mainly analyzed.Import the experimental data into Excel 2016 for preliminary statistical. Significance analysis was performed using SPSS (version 26.0). Non-parametric tests  were used to compare the differences in milk composition, rumen fermentation indexes, and fatty acid composition between the two groups of cows . The omi3.3.1.p = 0.958). The CP, NDF, ADF, and DMI intakes of cows in HF group were significantly higher than that in the LF group (P<0.05). The milk fat percentage is the most concerned indicator. The milk fat percentage of the dairy cows in the HF group was 4.42, which was significantly greater than that in the LF group (p < 0.01).3.2.3-N concentration of rumen fluid between the two groups of dairy cows were similar, and the difference was not significant (p > 0.05). At the same time, the difference of total VFAs and individual VFAs between the two groups of dairy cows was also not significant (p > 0.05). However, it was worth noting that the concentration of VFAs in the rumen fluid of dairy cows in the HF group was higher than those in the LF group. In addition, the ratio of acetic acid to propionic acid and the mole percentage of each VFA between the two groups of dairy cows were not significant, and there was no statistical significance.The analysis results of the rumen fermentation indexes of dairy cows in the two groups were shown in 3.3.p < 0.05, p < 0.05, p < 0.01). It indicated that there is a higher abundance of bacteria in the HF rumen.16S rRNA gene sequencing analysis of rumen fluid samples from 16 cows obtained a total of 1,624,665 sequences, with an average of 10,1,541 ± 4,613 sequences per sample. After the sequencing results were processed for quality control, the bacterial richness index was obtained by using the alpha diversity analysis of rumen bacteria. The results in By listing species abundance information based on OTUs, a PCA plot was carried out to show the similarity between and within the two groups of dairy cows. The PCA diagram indicated that the variation degrees of the PC1 and PC2 were 56.67 and 21.93%, respectively. Moreover, the rumen bacterial composition of HF group and LF group dairy cows was clearly distinguished, indicating the rumen bacterial structure was significantly different between HF group and LF group . Samples3.4.Prevotella_1 was the highest, and both groups account for more than 20% of the total bacteria, followed by Succiniclasticum, and Prevotella-7. LF had 1.69 times more Succiniclasticum than HF , Prevotella_7 was 3.14 times higher than HF .A total of 25 bacterial phyla were discovered at the phylum level after classification analysis. Among them, the relative abundance of Bacteroidetes and Firmicutes accounted for more than 93.0% . The ratPrevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03 of rumen fluid in HF dairy cows were significantly greater than those in LF dairy cows (p < 0.05).There were 8 different genera of bacteria between the two groups, which basically belong to Bacteroidetes and Firmicutes . The rel3.5.Prevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03 bacterial relative abundance were significantly positive correlated with milk fat percentage, and all of them were negatively correlated with non-dairy solids and lactose content. Among the rumen fermentation indexes, the relative abundance of 8 rumen differential bacteria were all positively related to pH, but inversely correlated with NH3-N. The relative abundance of Prevotellaceae_UCG-003 was positively related to VFAs.Correlations between milk composition, rumen fermentation indexes, and relative abundance of rumen bacteria were analyzed in HF and LF dairy cows using Spearman rank correlation analysis. The result is shown in 3.6.p < 0.05) between the two groups of cows (p < 0.05) in cows in the high milk fat group than in cows in the low milk fat group. In contrast, cows in the high milk fat group had C16:1 c7, C17:1 t10, C18:1 c9, C18:3 c6, c9, c12, C20:1 c11, C20:2 c11, c14, C22:0, C22:1 c13, C20:3 c11, c14, c17, C20:4 c5, c8, c11, c14, C20:5 c5, c8, c11, c14, c17, C24:0, C22:4 c7, c10, c13, c16, and C22:5 c7, c10, c13, c16, c19 were significantly lower (p < 0.05) than cows in the low milk fat group. SCFA, LCFA, SFA, MUFA, PUFA, and Trans contents were not significantly different (p > 0.05) between the two groups.A total of 22 fatty acids were significantly different ; the relative abundance of Ruminococcus_1 was positively related to C18:1 t9 (p < 0.05); the relative abundance of Lachnospiraceae_XPB1014_group was positively related to C7:0 (p < 0.05); The relative abundance of Lachnospiraceae_AC2044_group was positively related to C18:1 t9 and C18:1 t11 (p < 0.05); The bacterial abundance of U29-B03 was positively correlated with the C15:0 iso (p < 0.05). Summarily, these 8 differential bacteria were positively correlated with short and medium chain fatty acids. However, it was inversely correlated with long chain fatty acids.Detecting fatty acids in raw milk by GC–MS, we determined a total of 59 fatty acids, and carried out correlation research with fatty acids. The result was shown in 4.3-N concentration indicated that the rumen maintained a stable acid–base environment, and the molar percentage of VFAs composition indicated that the rumen fermentation model of the dairy cows did not change. And, no differences were found in rumen fermentation indicators between HF and LF group dairy cows. This was difficult to explain the difference in milk fat between the two groups of cows. But research has found that the NDF and ADF feed intake of HF was significantly higher than that in LF dairy cows . Plant fibers are decomposed by rumen bacteria into VFAs and other precursors for synthetic milk fat  can be found at: The animal studies were approved by Animal Care Committee of Institute of Animal Sciences of CAAS . The studies were conducted in accordance with the local legislation and institutional requirements. Written informed consent was obtained from the owners for the participation of their animals in this study.BS: investigation, data curation, and writing – original draft. KL and GH: methodology and validation. MC and XW: formal analysis. JY, NL, and WT: investigation. YZ: conceptualization and writing – review. SZ: conceptualization and project administration. NZ: investigation and writing – review and editing. JW: supervision, conceptualization, and writing – review and editing. All authors contributed to the article and approved the submitted version.