LWT - Food Science and Technology 168 (2022) 113907

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Construction of ZnO@mSiO2 antibacterial nanocomposite for inhibition of

microorganisms during Zea mays storage and improving the germination
Dong-Dong Zhang 1, Si Hu 1, Qiong Wu , Jin-Feng Zhao , Ke-Rui Su , Li-Qin Tan ,
Xian-Qing Zhou *
Grain Storage and Security Engineering Research Center of Education Ministry, School of Food and Strategic Reserves, Henan University of Technology, Zhengzhou,
450001, PR China

A R T I C L E I N F O A B S T R A C T

Keywords: Zea mays (Maize) is prone to mildew due to its rich nutrition and large embryo, resulting in the decline of edible
ZnO nanoparticles quality, germination rate and even threatening people’s health. In this study, ZnO nanoparticles were prepared
ZnO@mSiO2 nanocomposite by hydrothermal method to inhibit the growth of microorganisms on the surface of Zea mays. For its safe and
Antimicrobial
efficient application, mesoporous silica (mSiO2) was modified on its surface and successfully prepared
Zea mays storage
ZnO@mSiO2 Nanocomposites. The ZnO@mSiO2 nanocomposites were successfully prepared and characterized
Safety
by TEM, EDS, XRD, FT-IR, and it could significantly inhibit two model bacteria and three model fungi. After
mixing with Zea mays and 42 days of simulated storage, they could significantly inhibit the growth of micro­
organisms in Zea mays, ensure the stability of Zea mays quality and improve the germination rate of Zea mays.
The ZnO@mSiO2 nanocomposites prepared in this study provide a new idea for the safe storage of Zea mays.

1. Introduction excessive growth of microorganisms would directly reduce the germi­

As one of the largest grain in the world, Zea mays (maize) is rich in
nutrients. In addition, Zea mays embryo is relatively large, easy to absorb
moisture and carry large amount of bacteria, which is very easy to
promote the rapid growth and reproduction of microorganisms on the
surface of Zea mays and mildew (Mannaa & Kim, 2017). If the moisture
content in Zea mays is high after harvest, in the process of transportation
and storage, coupled with the continuous metabolism of Zea mays itself,
it would provide a suitable growth environment for microorganisms on
the surface of Zea mays and cause the heat of corn pile, so as to further
promote the growth and reproduction of microorganisms and cause
safety problems such as the corruption, deterioration, mildew and
excessive mycotoxin (Li et al., 2021; Rodríguez-Félix et al., 2021; Wang,
Liu, Guo, He, & Lu, 2020). In addition, the massive reproduction of
microorganisms would destroy the color, taste, nutrition and processing
characteristics of Zea mays. The mycotoxins metabolized include afla­
toxin, deoxynivalenol, zearalenone, ochratoxin and so on, which are
recognized as carcinogens, which directly threaten human life and
health (Walker, Jaime, Kagot, & Probst, 2018; Wawrzyniak, Waskie­
wicz, & Ryniecki, 2018). During the storage of Zea mays seeds, the
nation rate of Zea mays seeds and make Zea mays seeds lose their value.
Zinc oxide (ZnO) nanoparticles are a kind of antibacterial materials
with broad spectrum, enduring effect, stable, low toxicity, and no special
requirements for the environment (Devi & Maji, 2012; Duan et al.,
2021). Therefore, ZnO nanoparticles have been widely used in medical
textile and farming applications such as cosmetics, decontamination,
protective medical clothing, packing bag and pig feed (Gudkov et al.,
2021; Seo et al., 2018; H. Zhang et al., 2018). Different synthesis
methods of nanoparticles will affect the shape, particle size and
dispersion of nanoparticles, which would directly affect the antibacterial
properties of ZnO nanoparticles (Rodríguez-Félix et al., 2022). In the last
few years, various methods have been used to prepare ZnO nano­
particles with different morphologies, including solvent-based ultra­
sonic irradiation (Pintaric et al., 2020), sol-gel method (Kumar, 2020) or
green methods such as plant extracts/bacterial strains (Dobrucka &
Dugaszewska, 2016; Ekennia et al., 2021; Ifeanyichukwu, Fayemi, &
Ateba, 2020). However, our hydrothermal synthesis method was not
only low cost, high efficiency, high product purity and good dispersion,
but also could prepare ZnO nanoparticles with small and uniform par­
ticle size. In the application of ZnO nanoparticles, there was a concern

* Corresponding author. No. 100 Lianhua Street, Zhengzhou High-Tech Development Zone, Henan, 450001, PR China.
E-mail address: xianqingzh@163.com (X.-Q. Zhou).
1
These authors contributed equally to the work.

https://doi.org/10.1016/j.lwt.2022.113907
Received 11 April 2022; Received in revised form 18 August 2022; Accepted 24 August 2022
Available online 28 August 2022
0023-6438/© 2022 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
D.-D. Zhang et al.
about the migration and direct contact effects of ZnO nanoparticles, for
example it may affect the activity of enzymes and protein structure in
Zea mays (Franz, Bott, & Stormer, 2020; Ricardo, Garcia, Bernardo, Rios,
& Martin-Doimeadios, 2021). Therefore, in order to avoid this, we added
a modified layer on the surface of ZnO nanoparticles so that ZnO
nanoparticles did not direct contact with Zea mays and play its anti­
bacterial role at the same time. Meanwhile, it could also avoid the ag­
gregation of ZnO nanoparticles and increase their dispersion.
Mesoporous silica (mSiO2) not only has large pore volume, adjustable
pore size, large specific surface area, good hydrophilicity, nonspecific
surface groups, but also has low biotoxicity, good biocompatibility,
chemical stability and its been applied in living organisms (Wu, Yang, &
Yan, 2017; Yue, Sun, Kang, & Deng, 2020). It is an ideal surface modi­
fication layer.
In this study, ZnO nanoparticles prepared by hydrothermal method
were used as the core of antibacterial particles, after the characterization
of morphology, structure and chemical bond, we modified a layer of
mesoporous silica on the surface of ZnO nanoparticles to avoid direct
contact with Zea mays. After confirming the morphology and structure of
the prepared ZnO@mSiO2 nanocomposites, the two model strains of
bacteria and three model strains of fungi were used to test the antimi­
crobial activity of the prepared ZnO@mSiO2 nanocomposite. Finally,
after 42 days of simulated storage with Zea mays, the inhibitory effect of
the prepared ZnO@mSiO2 nanocomposites on microorganisms during
Zea mays storage was tested. Meanwhile, the quality and germination
rate of stored Zea mays should not be destroyed. The brief process of the
study was shown in Fig. 1. We expect that the prepared ZnO@mSiO2
nanocomposites could be used to inhibit the growth of microorganisms
during Zea mays storage, and ensure the stability of Zea mays quality and
improve the germination rate of Zea mays.
2. Materials and methods
2.1. Chemicals and materials
Zinc acetate dehydrate (Zn(CH3COO)2⋅2H2O; 99.0%) (was pur­
chased from Shanghai Aladdin Biochemical Technology CO., Ltd,
China); sodium hydroxide (NaOH; 99.6%), Tetraethyl orthosilicate
(TEOS; 98.0%) (were purchased from Tianjin kermi ou chemical reagent
CO., Ltd, China); Cetyltrimethylammonium Bromide (CTAB; 99.0%)
(was obtained from Tianjin guangfu fine chemical research institute,
China). All the chemicals used in this study were at least analytical
grade, all the water used in this study was distilled twice, all strains used
in this study were preserved in our laboratory.
2.2. Characterization
The morphology of prepared nanoparticles was analyzed by a JEM-
2100F field emission transmission electron microscope (TEM, JEOL,
Fig. 1. Schematic illustration of the synthesis and surface modification of ZnO and their application in inhibiting the growth of bacteria and fungi and safe storage
of maize.
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LWT 168 (2022) 113907
Japan) operating at acceleration voltage of 200 kV, along with an energy
dispersive spectrometer (EDS). Kα-rays from a d8 advance X-ray poly­
crystalline diffractometer (XRD) were used to bombard the Cu target at
the wavelength of 1.5406 Å (Mydeen, Kumar, Kottaisamy, & Vasantha,
2020). The particle size distribution was determined by dynamic light
scattering (DLS) laser particle size analyzer (ZMV2000, Malvern Pan­
alytical Ltd, United Kingdom) (Cao, Gong, Shu, Zhu, & Liang, 2019). The
chemical bond structure was analyzed by ARffinity-1S Fourier transform
infrared spectrometer (Optosky Photonics Inc, Xiamen, China) with the
range of 4000-400 cm− 1 and a resolution of 1 cm− 1, the infrared spec­
trogram of nanomaterial powder was determined by conventional KBr
press method (Delgado-Licona, Lopez-Guajardo, Gonzalez-Garcia,
Nigam, & Montesinos-Castellanos, 2020). The specific surface area and
pore size were determined by surface adsorption instrument (Micro­
meritics Instrument Corporation, USA) (X. Wang et al., 2017).
2.3. Preparation of ZnO nanoparticles
The ZnO were synthesized by hydrothermal method. Firstly, 3 g of Zn
(CH3COO)2⋅2H2O was dissolved in 75 mL double distilled water and
stirred for 15 min. Meanwhile, 0.3 g CTAB was dissolved in 10 mL
double distilled water and stirred for 10 min. Then, mix CTAB solution
and zinc acetate solution, and the mixture was stirred for 15 min. Sec­
ondly, 2.4 g of NaOH was dissolved in 75 mL double distilled water,
stirred for 15 min, and added to the mixture solution prepared in the first
step, stirred for 30 min at 45 ◦ C. After ultrasonic treatment (ultrasonic
frequency 40 kHz, KQ-600KDE, Kunshan Ultrasonic Instrument Co.,
LTD, China) for 15 min, the mixture was transferred into the Teflon-
lined stainless steel autoclave and kept 120 ◦ C for 6 h. After cooled to
room temperature naturally (about 2 h), the harvested precipitates were
centrifuged (8000 g, 20 min, 4 ◦ C) and washed with double distilled
water and anhydrous ethanol three times. Finally, the prepared pre­
cipitate was vacuum freeze-dried to obtain the ZnO nanoparticles
powder.
2.4. Preparation of ZnO@mSiO2
The prepared ZnO nanoparticles as the core of ZnO@mSiO2. The
harvested ZnO nanoparticles (100 mg) were suspended in 100 mL
double distilled water with 0.5 mM NaOH, treated with ultrasonic bath
(ultrasonic frequency 40 kHz) for 5 min, 0.4 g CTAB was added to the
suspension and stirred at 45 ◦ C; 1.5 mL 10% TEOS/methanol (v/v) was
added drop by drop, stirring for 90 min at 45 ◦ C. Then, another 1.5 mL
10% TEOS/methanol (v/v) was added drop by drop, stirring for another
90 min at 45 ◦ C. The mixture was transferred to a Teflon-lined stainless
steel autoclave and kept 120 ◦ C for 24 h. After cooled to room temper­
ature naturally, the harvested compound was washed with double
distilled water and ethanol three times. To remove CTAB, the product
was well dispersed in 1% NaCl/methanol (m/v) with continuous
D.-D. Zhang et al.
stirring, and the solution was changed every 3 h, replace the solution
repeatedly for at least six times. At last, the final product was vacuum
freeze-dried to obtain the ZnO@mSiO2 nanocomposites powder.
2.5. Determination of inhibition circle
The plate counting method (colony forming unit analysis method)
was used to determine the antibacterial activity of ZnO@mSiO2. The
microorganisms used in this study were isolated and identified from the
surface of Zea mays, used as the model bacteria and fungi. The bacteria
were Escherichia coli and Staphylococcus aureus, the fungi were Aspergillus
flavus、Aspergillus Niger and Penicillium citrinum. For the detection of
inhibiting bacteria, ZnO@mSiO2 was mixed well with liquid lysogeny
broth (LB) medium with the concentrations of 0, 0.1, 0.25, 0.5, 0.75 and
1 mg/mL. Subsequently, 100 μL bacterial suspension (106 CFU/mL) was
spread evenly on LB agar plates, incubated at 37 ◦ C for 24 h (Zhao, Du,
et al., 2020).
For the detection of mold inhibition, ZnO@mSiO2 was mixed well
with potato dextrose agar (PDA) medium at concentrations of 0, 0.10,
0.25, 0.50, 0.75 and 1.00 mg/mL, and 100 μL fresh spore suspension
(106 CFU/mL) was spread evenly on PDA medium (Pariona et al., 2020).
After incubation at 30 ◦ C for 72 h, the average diameter of fungal radial
growth was measured with digital vernier calipers. The growth inhibi­
tion rate of the control (the addition concentration of ZnO@mSiO2 is 0
mg/mL) was calculated according to Equation (1).
C− T
Inhibition ​ of ​ mycelial ​ growth ​ (%) ​ = ​ × 100 (Equation 1)
C
Note: C is the mycelial length of the control group, and T is the
mycelial length of ZnO@mSiO2 treatment, in mm.
2.6. Determination of minimum inhibitory concentration
The resuspended cells (bacteria or spores of molds) in the medium
were prepared at the number of 106 CFU/mL, evenly mixed with
ZnO@mSiO2 of different concentrations. The concentrations used for
bacteria were 0–1000 μg/mL, fungi were 0–500 μg/mL. Subsequently,
incubated at 37 ◦ C for 12 h for bacteria and 30 ◦ C for 24 h for fungi. In
the medium mixed with ZnO@mSiO2, the bacteria or mold do not grow,
no visible strain was found on LB agar (or PDA agar) plates by spread
plate method. This minimum concentration of ZnO@mSiO2 was taken to
be the minimum inhibitory concentration (MIC) (Ji et al., 2020).
2.7. Inhibition of microbial growth during Zea mays storage
The prepared ZnO@mSiO2 was suspended in sterile water and
dispersed by ultrasound (ultrasonic frequency 40 kHz) for 30 min. The
suspension was evenly sprayed on the surface of Zea mays, and the Zea
mays was rotated (100 r/min) and stirred in a Humidification wheat
(BLH-1780, Zhejiang Bethlehem Apparatus Co., Ltd., China) for 30 min,
make sure the concentration of ZnO@mSiO2 in Zea mays was 1500 mg/
kg. The sterile water without ZnO@mSiO2 was used as control, and the
same treatment. The treated Zea mays was stored at 30 ◦ C and 80%
humidity, the microbial changes in Zea mays were detected every 7 days.
2.8. Zea mays storage quality detection
The determination method of fatty acid value of Zea mays was based
on Chinese National Standard (GB/T 20570-2015 Guidelines for eval­
uation of maize storage character), the basic steps are as follows. Took
10.00 g ground Zea mays sample, added 50 mL ethanol, and shook for
30 min. Took 25 mL filter liquor, added 50 mL of distilled water and 5
drops of phenolphthalein solution, and titrated with potassium hy­
droxide standard titration solution to reddish.
The determination method of germination rates of Zea mays was
based on Chinese National Standard (GB/T 5520-2011 Inspection of
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LWT 168 (2022) 113907
grain and oils-Germination test of seeds), the basic steps are as follows.
Placed 1 cm of fine sand in the Petri dish and added water to saturation.
Randomly selected 50 Zea mays seeds and place them in the Petri dish,
with the two seeds alternating 1–2 times the Zea mays length. These Zea
mays were cultivated at 30 ◦ C for 7 days.
2.9. Statistical analysis
All experiments were performed in triplicate, and the means of
replicates’ values were used for figures in this study. The least significant
difference (LSD) computations were performed to determine significant
differences between different treatments. The statistical analyses were
conducted by SPSS 17.0 software, and the differences were considered at
the 0.95 or 0.99 confidence level.
3. Results and discussion
3.1. Successful preparation of ZnO and ZnO@mSiO2
3.1.1. Morphological and EDS characterization
The prepared nanoparticles of ZnO and ZnO@mSiO2 were white
powder, loose and easy to disperse (Fig. 2A). Under the visual field of
TEM, ZnO nanoparticles were spherical, with irregular edges and good
dispersion (Fig. 2B). one hundred ZnO nanoparticles were selected from
the TEM images, and the average particle size was determined to be 33
± 2 nm, which was also the real particle size of ZnO nanoparticles.
Under the visual field of TEM, the ZnO@mSiO2 nanocomposites were
spherical and some of them gather together (Fig. 2C). There was an
obvious coating on the surface of ZnO nanoparticles, after mesoporous
silica modification, the dark ZnO core and light mesoporous silica
coating are clearly visible, which due to the high-energy electron beam
passes through different thicknesses of the nanocomposite. The thick­
ness of mSiO2 modified layer was about 10 nm. ZnO core has high
density, when it was collided by high-energy electron beam, the scat­
tering angle could be significantly affected. However, the peripheral
modified mSiO2 layer had a relatively small scattering angle due to its
low density and uncompact structure, thus leading to the different bright
and dark images of ZnO@mSiO2 (Cao et al., 2019).
The spectra of EDS showed that the prepared ZnO nanoparticles
contained only Zn and O elements (Fig. 2D), and the elemental mapping
of ZnO (Fig. 2E) showed that the Zn atoms and O atoms were uniformly
distributed. The spectra of EDS showed that the prepared ZnO@mSiO2
nanocomposites contained only Zn, Si and O elements (Fig. 2G). and the
elemental mapping of ZnO@mSiO2 (Fig. 2H) showed that the Zn atoms,
O atoms and Si atoms were uniformly distributed. There were no
obvious impurity elements in the specturm, which showed that the
elemental composition of the prepared samples were relatively pure.
3.1.2. Particle size distribution
The particle size range of ZnO@mSiO2 determined by dynamic light
scattering (DLS) method in this study was 200 nm – 460 nm, with an
average particle size of 268 nm (PDI = 0.081) (Fig. 2F). However, the
particle size range of ZnO determined by the DLS method was 68 nm –
122 nm, with an average particle size of 93 nm (PDI = 0.024) (Fig. 2F),
indicated that after the surface modification by mSiO2, the particle size
of ZnO nanoparticles increased significantly. The modified layer could
not only directly increase the diameter of ZnO nanoparticles, but also
caused the aggregation of some ZnO nanoparticles, which also increased
the particle size in the detection of nanoparticles. Meanwhile, the
determined particle sizes of ZnO or ZnO@mSiO2 were larger than the
results by TEM method, it due to the nanoparticles would collide with
the surrounding particles in the solution while doing Brownian move­
ment, it even combined with –OH groups to drive the movement of
surrounding water molecules, resulting in the large-scale simultaneous
movement of particles in the detection process (Zhang et al., 2019).
D.-D. Zhang et al. LWT 168 (2022) 113907

Fig. 2. Characterization of the prepared ZnO and ZnO@mSiO2. (A) Image of the prepared ZnO@mSiO2. (B) TEM images of the prepared ZnO. (C) TEM images of the
prepared ZnO@mSiO2. (D) EDS spectrum of the prepared ZnO. (E) Elemental mapping of the prepared ZnO. (F) Size distributions of the prepared ZnO and
ZnO@mSiO2. (G) EDS spectrum of the prepared ZnO@mSiO2. (H) Elemental mapping of the prepared ZnO@mSiO2. (I) The XRD patterns of the prepared ZnO and
ZnO@mSiO2 compared with standard PDF cards. (J) FTIR spectra of the prepared ZnO and ZnO@mSiO2. (K) Nitrogen adsorption and desorption isotherms of the
prepared ZnO and ZnO@mSiO2. (L) Pore-size distributions of the prepared ZnO and ZnO@mSiO2.

3.1.3. Structural characterization
The crystal structure of the prepared ZnO and ZnO@mSiO2 nano­
composites was confirmed by XRD (Fig. 2I). The spectrum showed a
series of characteristic peaks of ZnO, which was consistent with the zinc
ore of ZnO standard spectrum (PDF#01-070-2551, International
Diffraction Data Center, JCPDS5-0664) (Zhai, Tao, Pu, Zeng, & Chen,
2010). There were no impurity peaks in the spectrum, indicating that the
structure of the prepared ZnO nanoparticles was pure and complete.
After the modification of mSiO2 coating layer, the characteristic peaks of
ZnO@mSiO2 nanocomposites include the standard characteristic
spectra of zinc oxide (PDF#01-070-2551) and SiO2
(PDF#01-070-8054), the characteristic peak of SiO2 appears obviously,
compared with the XRD spectrum of ZnO. The results indicated that
mSiO2 layer had no effect on the crystal structure of ZnO and was coated
on the surface of ZnO nanoparticles successfully.
The vibrational bands induced by Zn–O bonding and the changes
induced by the modification of Si–O bonds were analyzed by fourier
transform infrared spectroscopy (FTIR), the typical FT-IR absorption
spectra of prepared ZnO and ZnO@mSiO2 nanocomposites were showed
as Fig. 2J. There was a medium peak at 3500 cm− 1, which might be
caused by the –OH group coordinated to the zinc ion or due to the water
on the surface of the nanoparticles. The stretching and bending vibra­
tional absorption peaks at 1629 cm− 1 and 472 cm− 1 were the charac­
teristic absorptions peaks of Zn–O bond (Fig. 2J). The peaks at 3380
cm− 1 and 1625 cm− 1 may be due to –OH or water absorption in the
ZnO@mSiO2 nanocomposites sample, and the characteristic peaks at

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D.-D. Zhang et al.
1000 cm− 1, which must be attributed by the typical Si–O–Si asymmetric
stretching vibrations (Zafar et al., 2019). It showed that the mSiO2 shell
formed by the hydrolytic condensation of TEOS was successfully coated
on the surface of ZnO nanoparticles.
3.1.4. The pore size and specific surface area
The specific surface area of ZnO@mSiO2 reached 162.56 m2/g,
detected by BET (Brunauer-Emmett-Taylor) method, which was signif­
icantly higher than that of ZnO (Fig. 2K). The average pore size of
ZnO@mSiO2 was 5.27 nm, which was calculated by BJH (Barrett-Joy­
ner-Halenda) method, the distribution curve of pore volume and pore
size were also clearly shown in Fig. 2L. According to the definition of
nitrogen adsorption and desorption isotherm, the pore structure could
be divided into three types: macroporous (r > 50 nm), mesoporous (2
nm < r < 50 nm) and microporous (r < 2 nm), which indicated that the
prepared ZnO@mSiO2 was typical type IV mesoporous materials with
good nitrogen adsorption effect (Yue et al., 2020). Meanwhile, this also
indicated that the mesoporous silica was better modified on the surface
of ZnO nanoparticles, and the specific surface area of ZnO nanoparticles
was significantly improved.
3.2. Inhibition of microbial growth by prepared ZnO@mSiO2
3.2.1. Two strains of model bacteria
We selected two strains of bacteria, Staphylococcus aureus (S. aureus)
and Escherichia coli (E. coli), isolated and identified from the surface of
Zea mays, used as the model bacteria for the study of bacteriostasis of
ZnO@mSiO2. With the increase of the concentration of ZnO@mSiO2, the
total number of S. aureus and E. coli colonies in the medium plate
decreased (Fig. 3). When the concentration of ZnO@mSiO2 was 0.1 mg/
mL, the total number of S. aureus colonies was significantly less than that
of E. coli colonies. When the concentration of ZnO@mSiO2 was 0.25 mg/
mL, there was almost no growth of S. aureus in the medium plate, and the
total number of E. coli colonies also significantly decreased. Therefore,
with the increase of the concentration of ZnO@mSiO2 nanocomposites,
ZnO@mSiO2 could kill almost all S. aureus, while the concentration
Fig. 3. Inhibition of five model strains on medium plate by ZnO@mSiO2. The five model strains are two strains of bacteria, Escherichia coli (E. coli), Staphylococcus
aureus (S. aureus), three strains of fungi, Aspergillus flavus (A. flavus), Aspergillus niger (A. niger) and Penicillium citrinum (P. citrinum), which were all isolated
from maize.
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LWT 168 (2022) 113907
required to kill all E. coli was a little higher. The results showed that the
ZnO@mSiO2 nanocomposites could significantly inhibit the growth of
two model bacteria, which was similar to the results of Liu’s study (Liu
et al., 2018). It was shown that the growth inhibitory properties of ZnO
nanoparticles would be accompanied by the loss of corresponding cell
viability. Many previous studies (Hermida-Montero et al., 2021; Ilkhe­
chi, Mozammel, & Khosroushahi, 2021; Vazquez et al., 2021; Y. Zhang,
Zhou, Cheng, Luo, & Sun, 2021) have also confirmed that ZnO nano­
particles in ZnO@mSiO2 could generate reactive oxygen species (ROS)
free radicals, such as superoxide and hydroxyl, which could change the
activity of membrane proteins or enzymes, resulting in the damage of
cell membrane, and cell death. Meanwhile, Zinc ions and ROS could
enter bacteria to inhibit the replication of bacterial DNA, and inhibit the
expression of protein and enzyme genes in bacteria. Zinc ions could also
interact with ribosomal proteins to inhibit the production of bacterial
proteins, this would inhibit the growth of bacteria.
The minimum inhibitory concentration (MIC) is an effective method
for quantifying the antibacterial activity of ZnO@mSiO2 nano­
composites, and MIC was the lowest concentration of ZnO@mSiO2
nanocomposites that resists the growth of a microorganism in medium
(El-Borady & El-Sayed, 2020). The minimum inhibitory concentration of
ZnO@mSiO2 nanocomposites against S. aureus and E. coli were 0.5
mg/mL and 0.9 mg/mL, respectively (Fig. 4A). As a gram-negative
bacterium, E. coli has higher minimum inhibitory concentration. This
finding confirmed that gram-positive bacteria were more susceptible to
ZnO@mSiO2 nanocomposites compared with Gram-negative bacteria.
This has been confirmed by many studies (Alam et al., 2022; Elumalai,
Velmurugan, Ravi, Kathiravan, & Ashokkumar, 2015; Yusof, Zain, &
Pauzi, 2019), which due to the possibilities of membrane damage were
caused by electrostatic interaction between ZnO and cell surface.
3.2.2. Three strains of model fungi
We selected three strains of fungi, Aspergillus flavus (A. flavus),
Aspergillus niger (A. niger) and Penicillium citrinum (P. citrinum), isolated
and identified from the surface of Zea mays, used as the model fungi for
the study the inhibition of ZnO@mSiO2 on the growth of fungal hyphae.
D.-D. Zhang et al. LWT 168 (2022) 113907

Fig. 4. Effects of different concentrations of ZnO@mSiO2 on bacterial (A) and fungal (B).

Different concentrations of ZnO@mSiO2 nanocomposites showed
different inhibitory effects on three different fungi, the higher the con­
centration of ZnO@mSiO2 nanocomposites, the more obvious the inhi­
bition of mycelial growth (Fig. 3, Table 1), probably due to the different
mycelial morphology of these three fungi led to different antifungal
effects. The inhibition of ZnO@mSiO2 nanocomposites on the growth of
A. niger was the most significant, when the concentration was 1 mg/mL
and cultured for 72 h, the inhibition rate of ZnO@mSiO2 nano­
composites on the mycelium was 72.89 ± 1.46%. P. citrinum was the
least sensitive, under the same treatment conditions, the inhibition rate
was 44.18 ± 1.53%. This probably due to different fungi have different
active defense mechanisms against ROS, resulting in the structural
damage of fungal cells and the inhibition of conidial germination (Sun
et al., 2019; Zhao, Chen, et al., 2020). Meanwhile, ZnO@mSiO2 nano­
composites could affect the function of mycelial cells, deform mycelium,
or inhibit the development of conidia, resulting in the inhibition of
fungal growth.
Different concentrations of nanoparticles were cultured with
A. flavus, A. niger and P. citrinum for 36 h, the measured MIC were as
follows: the MIC of A. flavus was 0.5 mg/mL, and the MIC of both A. niger
and P. citrinum was 0.4 mg/mL (Fig. 4B). Some researchers confirmed
that the antimicrobial potential of ZnO nanoparticles was due to their
unique properties such as disruption of cell walls and cell membranes,
prevention of cell transport and leakage of internal cell contents (Ahmed
et al., 2021; Korshed, Li, Ngo, & Wang, 2018).
3.3. Inhibition of total bacterial count in stored Zea mays by prepared
ZnO@mSiO2
the storage period, fluctuating around 150 CFU/mL (Fig. 5A). The pre­
pared ZnO@mSiO2 significantly inhibited the growth of E. coli and
S. aureus (Fig. 3), and the total number of bacterial colonies on the Zea
mays surface was significantly reduced during the storage period, indi­
cating that it must significantly inhibited the bacterial colonies of other
species on the Zea mays surface, and could be applied to the inhibition of
bacterial growth during the storage of Zea mays. As it was shown in
Fig. 5A, the total number of bacterial colonies in the ZnO@mSiO2
treatment group was consistently lower than the control group, indi­
cating that ZnO@mSiO2 nanocomposites had a sustained inhibitory ef­
fect on bacteria, and the addition of ZnO@mSiO2 nanocomposites to Zea
mays could inhibit the growth of bacteria during Zea mays storage for a
long time.
3.3.2. The total number of fungal colonies
After 42 d of storage at 30 ◦ C and 80% relative humidity, the total
number of fungal colonies on the surface of Zea mays in the ZnO@mSiO2
treatment group was relatively stable during the storage, fluctuating
around 5000 CFU/mL. However, the total number of fungal colonies in
the control group increased significantly, with the total number of
fungal colonies increasing from the initial 4500 CFU/mL to 51500 CFU/
mL (Fig. 5B). This indicated that ZnO@mSiO2 nanocomposites could not
only inhibit the growth of bacteria, but also inhibit the growth of fungi,
which it was more destructive during Zea mays storage. Meanwhile, the
total number of fungal colonies in the ZnO@mSiO2 treatment group was
consistently lower than that in the control group during the storage
period of 42 d, and it was remained at a low level, indicated that the
addition of ZnO@mSiO2 had a good long-term inhibitory effect on the
total number of fungal colonies.

3.3.1. The total number of bacterial colonies 3.4. Effect on fatty acid value of Zea mays during storage
After 42 d of storage at 30 ◦ C and 80% relative humidity, the total
number of bacterial colonies on the surface of Zea mays in the control Fatty acid value was an important indicator of Zea mays during
group increased significantly (P < 0.01) with the extension of storage storage, its value represents the deterioration degree of Zea mays quality
time (Fig. 5A), and the total number of bacterial colonies increased from and was an important indicator of whether Zea mays was suitable for
initially 950 CFU/mL to 10500 CFU/mL. However, when ZnO@mSiO2 storage. After 42 d of storage, the fatty acid values of Zea mays in the
nanocomposites with a concentration of 1.5 g/kg were added to Zea control group increased from the initial 36.94 mg/100 g to the final
mays, the total number of bacterial colonies on the surface of Zea mays 45.28 mg/100 g, and those of Zea mays in the ZnO@mSiO2 treatment
was significantly inhibited, and it was remained relatively stable during group increased from the initial 34.46 mg/100 g to the final 43.00 mg/
100 g (Fig. 6A). The fatty acid values of Zea mays increased significantly
Table 1 (p < 0.05) with the increase of storage time, and the addition of
Growth inhibition (%) of the three fungal mycelial growth treated by ZnO@mSiO2 did not show any significant difference in the fatty acid
ZnO@mSiO2 values compared to the control group. On the one hand, it also
Concentration (mg/ Inhibition of mycelial growth (%) confirmed that the addition of ZnO@mSiO2 would not cause damage to
mL) the fatty acid content of Zea mays. On the other hand, it indicated that
Aspergillus Aspergillus Penicillium
niger flavus citrinum the fatty acid values of Zea mays did not change much during the shorter
0.10 51.60 ± 0.93 25.99 ± 1.06 23.13 ± 1.06 storage period, and its quality deteriorates slowly.
0.25 54.75 ± 0.99 41.14 ± 1.50 28.05 ± 0.50
0.50 63.74 ± 0.76 53.49 ± 1.56 33.11 ± 1.53 3.5. Effect on germination rate of Zea mays during storage
0.75 69.20 ± 1.04 59.91 ± 1.10 37.74 ± 0.87
1.00 72.89 ± 1.46 64.71 ± 1.06 44.18 ± 1.53
On the one hand, the germination rate of Zea mays was related to the

6
D.-D. Zhang et al. LWT 168 (2022) 113907

Fig. 5. Inhibition of the total number of bacterial (A) and fungal (B) colonies during maize storage.

Fig. 6. Effect on fatty acid value (A) and germination rate (B) of maize during storage. Maize germination photos of control group (C) and ZnO@mSiO2 treatment
group (D).

moisture content of seeds, it was a direct evidence of whether the Zea
mays embryo was intact. After 42 d of storage, the germination rate of
Zea mays in control group was maintained above 90% in the early stage
of storage, and there was a decrease in the late stage of storage, while the
germination rate of Zea mays in the ZnO@mSiO2 treatment group was
relatively higher, and the whole time period of them were higher than
that of the control group (Fig. 6B). It was indicated that ZnO@mSiO2
had some protective effect on the germination rate of Zea mays, which
might be due to that ZnO@mSiO2 inhibited the growth of microorgan­
isms during the storage of Zea mays, thus protecting the integrity of the
Zea mays embryo and promoting the germination rate (Chen et al., 2019;
dos Santos et al., 2021). Moreover, the ZnO@mSiO2 treatment group
had faster germination growth and stronger roots compared to the
control group (Fig. 6C/D), which indicated that the addition of
ZnO@mSiO2 could increase the rate of germination, probably due to the
fact that the ZnO@mSiO2 nanocomposites contained zinc element,
which could act as a nutrient to promote the growth of the germ.
4. Conclusions
In this study, we prepared ZnO nano antibacterial particles by hy­
drothermal method, and modified a mesoporous silica coating on the
surface of ZnO nanoparticles, successfully prepared ZnO@mSiO2
nanocomposites. TEM images showed that the prepared ZnO@mSiO2
nanocomposites were spherical with a particle size of 40±5 nm, EDS
spectrum shows that it did not contain impurity elements. XRD and FTIR

7
D.-D. Zhang et al.
spectrum shows that the structure of the prepared ZnO@mSiO2 nano­
composites was complete and consistent with the standard spectrum,
and it also showed that the mSiO2 coating was successfully modified on
the surface of ZnO nanoparticles. The nitrogen absorption/desorption
test confirmed the mesoporous structure of the prepared ZnO@mSiO2
nanocomposites, with a pore size of 5.27 nm. Two strains of bacteria and
three strains of fungi isolated from Zea mays were selected as model
strains, the antibacterial properties of the prepared ZnO@mSiO2 nano­
composites and the MIC against five strains were studied, the MIC of
S. aureus, E. coli, A. flavus A. niger, and P. citrinum were 0.5 mg/mL, 0.9
mg/mL, 0.5 mg/mL, 0.4 mg/mL and 0.4 mg/mL, respectively. With the
increase of ZnO@mSiO2 nanocomposites concentration, the antimicro­
bial effect on the five strains was also enhanced. The prepared
ZnO@mSiO2 nanocomposites were added to Zea mays to simulate stor­
age for 42 days, it could not only inhibit the growth of bacteria and
fungi, but also inhibit the deterioration of Zea mays quality.
The results showed that the prepared ZnO@mSiO2 could inhibit the
growth and reproduction of microorganisms on the surface of Zea mays,
and it was expected to be used as a new type of mildew inhibitor in the
storage process of Zea mays, to achieve long-term inhibition of the
growth and reproduction of microorganisms on the surface of Zea mays,
and to enhance the storage stability of Zea mays.
CRediT authorship contribution statement
Dong-Dong Zhang: Conceptualization, Software, Revised manu­
script preparation. Si Hu: Methodology, Data curation, Software,
Writing – original draft. Qiong Wu: Validation, Software. Jin-Feng
Zhao: Investigation, Data curation. Ke-Rui Su: Investigation. Li-Qin
Tan: Investigation. Xian-Qing Zhou: Supervision, Reviewing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
The data that has been used is confidential.
Acknowledgements
This work was supported by the National Natural Science Foundation
of China (Grant No. 32001745), Young Elite Scientists Sponsorship
Program by CAST (Grant No. 2019QNRC001), Supported by China
Agriculture Research System of MOF and MARA (Grant No. CARS-03).
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