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Effects of nanoparticles on the activity and resistance genes of anaerobic digestion enzymes in livestock and poultry manure containing the antibiotic tetracycline

  • Xiaofei Zhen EMAIL logo , Ruonan Jiao , Han Zhan , Wenbing Wu , Ke Li , Lei Feng and Tie Du
Published/Copyright: July 12, 2024
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Open Chemistry
From the journal Open Chemistry Volume 22 Issue 1

Abstract

Taking chicken manure containing antibiotic oxytetracycline (OTC) as the research object, nano-Fe2O3 with a concentration of 300 mg/kg TS and nano-C60 with a concentration of 100 mg/kg TS as carriers were used for a 55-day sequential batch mesophilic anaerobic digestion experiment. The gas production performance, chemical parameters, antibiotic content, enzyme concentration, and resistance gene changes during anaerobic digestion were analyzed to clarify the effect of nanoparticles on anaerobic systems containing antibiotic chicken manure and provide a reliable basis for pollution reduction capacity of livestock and poultry manure. The results showed that (1) adding nano-Fe2O3 and nano-C60 promoted anaerobic gas production in chicken manure with different concentrations of OTC. The cumulative gas production from days 1 to 10 was 2,234(T5) > 2,163(T4) > 1,445(T2) > 1,289(T3) > 1,220(T1) > 1,216(CK) mL. The cumulative gas production of T4 and T5 increased by 77.29 and 83.11%, respectively. The final cumulative gas production for each group was 3,712(CK), 3,993(T1), 4,344(T2), 4,447(T3), 4,671(T4), and 4,849(T5) mL. The final OTC residue concentrations were 15.25, 20.40, 56.56, 17.67, and 16.89 μg/L, with degradation rates of 98.31, 98.80, 98.29, 99.07, and 99.11% respectively; (2) antibiotic OTC increased the activities of dehydrogenase, amylase, protease, and urease, while adding nanoparticles increased the activities of dehydrogenase and amylase, with no significant impact on cellulase, urease, and protease activities; (3) antibiotic resistance gene (ARGs) of various types were found in groups T2, T4, and T5, with the multidrug resistance gene (ARGs)-Multidrug accounting for a high proportion of 33.54, 35.63, and 37.43%, respectively, while the proportions of the other four types MLS, tetracycline, glycopeptide, and peptide ranged from 8.18 to 12.98%.

Keywords: nanoparticles; oxytetracycline; anaerobic digestion; antibiotics; resistance genes

1 Introduction

The expansion of the population has increased the demand for animal-derived food, and industrialization and intensification have become important features of modern livestock farming. The concentrated discharge of large amounts of animal waste is causing increasingly severe environmental pollution problems [1]. According to the second national pollution source survey, the chemical oxygen demand emissions reached 21.44 million tons in 2017, with livestock excrement being the main source of pollution, contributing 46.67% [2]. The use of antibiotics in the livestock industry is a common phenomenon, with China’s total antibiotic use in 2013 estimated at around 162,000 tons, of which 52% were for veterinary use. Commonly used antibiotics include tetracyclines, sulfonamides, quinolones, macrolides, and β-lactams. Livestock farms discharge bacteria containing resistant genes into the environment through treated wastewater and solid waste. Antibiotics have biological toxicity, strong inhibitory effects on microbial activity, and may induce resistant bacteria and antibiotic resistance genes (ARGs). The transfer of resistance genes can occur between antibiotic-resistant bacteria and local bacteria through conjugation, transformation, and transduction. Antibiotic-resistant bacteria may enter the human body through various means, including consuming contaminated food or water, as well as through occupational exposure such as inhalation [3], posing a threat to human life and health [4,5,6,7].

Anaerobic digestion technology is becoming increasingly mature, which can effectively reduce, harmlessly treat, and resource livestock and poultry manure. The clean energy biogas produced can alleviate the pressure of today’s energy shortage, and utilizing livestock and poultry manure to produce biogas can meet 4–5% of China’s annual energy demand [8]. By starting a biogas project using livestock and poultry manure as raw materials, the antibiotic content and concentration have a certain impact on the gas production performance of anaerobic systems. Ke et al. [9] found that tetracycline at concentrations of 20, 50, and 80 mg/L caused reductions in biogas production from mesophilic anaerobic digestion of cattle manure by 43.83, 65.10, and 77.79%, respectively. Ma et al. [10] reported that lincomycin and chloramphenicol at 25 mg/L led to decreases in total methane production of 29.39 and 19.88%, respectively, further decreasing to 51.27 and 49.46% at a concentration of 50 mg/L. Mitchell et al. [11] demonstrated that tylosin at concentrations below 100 mg/L had no inhibitory effect on overall biogas production, but as the dosage was gradually increased to 130, 260, 520, and 913 mg/L, biogas yields decreased by 10, 20, 30, and 38%, respectively. Zhao et al. [12] observed that norfloxacin at 500 mg/kg inhibited methane production rate without affecting the total methane yield. According to Cetecioglu et al. [13], sulfamethoxazole at concentrations between 100 and 250 mg/L significantly suppressed gas generation. Nanoparticles refer to materials with at least one dimension within the 100 nm range or composed of nanostructural units in a three-dimensional space spanning the nanoscale (1–100 nm). Nanoparticles exhibit surface effects, characterized by a dramatic increase in surface area, surface energy, and surface tension as particle size decreases. This renders nanoparticles highly chemically reactive, making them prone to interaction with other chemical substances. In this context, this study focuses on chicken manure containing tetracycline antibiotics, using nano-Fe2O3 and nano-C60 as carriers for a 55-day batch anaerobic digestion experiment. By analyzing gas production performance, chemical parameters, antibiotic content, enzyme concentration, and resistance gene changes during the anaerobic digestion process, the impact of nanoparticles on antibiotic-containing chicken manure anaerobic systems is clarified. This research provides important insights for maintaining ecological integrity, improving energy structure, and promoting sustainable development of ecological organic agriculture.

2 Materials and methods

2.1 Experimental equipment

The experiment uses a self-designed anaerobic digestion potential test device, as shown in Figure 1. The test system consists of two wide-mouth bottles (1 L) and 1 capacity bottle (1 L), which serve as the raw material digestion tank, biogas collection bottle, and water collection bottle, respectively. They are connected by anti-aging treated rubber tubes to form a set of gas communication devices. When connecting the device, it is necessary to ensure air tightness.

Figure 1 Physical and schematic illustration of the apparatus: (1) water bath; (2) digestion-reaction bottle; (3) mixture of chicken manure and acclimated sludge; (4) air duct; (5) gas cylinder; (6) distilled water; (7) beaker.
Figure 1

Physical and schematic illustration of the apparatus: (1) water bath; (2) digestion-reaction bottle; (3) mixture of chicken manure and acclimated sludge; (4) air duct; (5) gas cylinder; (6) distilled water; (7) beaker.

2.2 Experimental materials

The chicken manure used in this experiment was taken from a certain meat chicken breeding base. The difficult-to-degrade substances such as eggshells, feathers, and stones in the chicken manure were picked out and stored in a refrigerator at 4°C for later use. It was tested that the chicken manure did not contain any antibiotics.

The anaerobic digestion inoculum used in the experiment comes from the residual sludge of the surrounding sewage treatment plant. The residual sludge is transferred to a large sealed plastic container, with a temperature of about 20°C during transportation. After returning to the laboratory, the residual sludge is domesticated under the condition of 37°C. The domestication method is to place 5 L of recovered activated sludge in a 25-L sealed plastic container and incubate at a constant temperature of 37°C for 3 days. After the adaptation period, 2.5 kg of chicken manure taken out in advance to return to room temperature is added to the activated sludge for a 10-day incubation. Then, 5 kg of kitchen waste returned to room temperature is taken for domestication and incubation for 10 days, completing the domestication. The characteristics of chicken manure and inoculated sludge are shown in Table 1.

Table 1

Properties of the chicken manure and the acclimated sludge [14]

Substance TS (%) VS (%) C (%) H (%) O (%) N (%) S (%) C/N
Chicken manure 27.32 24.49 47.338 6.256 43.786 1.981 0.639 23.896
Acclimated sludge 2.21 0.87 19.970 3.270 74.888 1.180 0.692 16.930

TS, total solids; VS, volatile solid.

2.3 Experimental methods

2.3.1 Reagents

Preparation of nano-Fe2O3 suspension: nano-Fe2O3 (90 nm) was purchased from Shanghai Maclin Biochemical Technology Co., Ltd., with a purity of 99.8%.

Preparation of nano-C60 suspension: Fullerene C60 was purchased from Shanghai Maclin Biochemical Technology Co., Ltd.: purity, 99.9%. Antibiotics were purchased from Shanghai Aladdin Co., Ltd.: Tetracycline (97%, CAS: 6153-64-6) – chemical formula, C22H24N2O9·2H2O; relative molecular weight, 496.46, purity ≥98%, and stored in a 4°C refrigerator before use.

2.3.2 Experiments

The study set up a total of 6 groups of experiments, each group adding 120 g of chicken manure and 300 g of inoculated sludge; the maximum residual concentration of oxytetracycline (OTC) was 47.25 mg/kg TS, approximately taken as 50 mg/kg TS, and added at 50, 100, and 200% of the maximum residual concentration, respectively. In experiments T4 and T5, under a concentration of 50 mg/kg TS of OTC, 300 mg/kg TS of nano-Fe2O3, and 100 mg/kg TS of nano-C60 were added. The materials for each group are as shown in Table 2. After the feeding, deionized water was added to reach a final volume of 1 L, followed by nitrogen stripping for 2 min to remove the air. The tank was then sealed. The anaerobic digestion reactor was placed in a thermostat water bath at 37 ± 0.5°C and shielded from light. Four replicates were set up for each group. One replicate was only used for supplementing the material lost for the sampling detection, and the final result of each group was the average of the remaining three replicates. Throughout the experiments, the slurry was stirred manually twice daily, for 1 min each time.

Table 2

Experimental design

Serial no. Fresh chicken manure (g) OTC (mg/kg TS) Nano-Fe2O3 (mg/kg TS) Nano-C60 (mg/kg TS) Initial concentration (μg/L) Amount of acclimated sludge (g)
CK 120 0 0 0 0 300
T1 120 25 0 0 700 300
T2 120 50 0 0 1,900 300
T3 120 100 0 0 3,300 300
T4 120 50 300 0 1,900 300
T5 120 50 0 100 1,900 300

2.4 Monitoring method

2.4.1 Analysis of physicochemical properties

The conventional indicators tested in this study mainly focus on the supernatant of the digestive fluid, including volatile fatty acids (VFAs) and pH value. The slurry supernatant was obtained by centrifugation of an appropriate amount of the digestion mixture using a high-speed refrigerated centrifuge at 9,000 rpm for 10 min, followed by filtration using a 0.45 μm filter membrane.

The specific testing method is as follows:

(1) VFAs: In this study, the composition of VFAs was determined using a gas chromatograph. The supernatant samples of sludge were further filtered through a 0.22-μm filter membrane, 1 mL of sample was added to the injection bottle along with 100 μL of chromatographic grade formic acid, and the sample was acidified to pH < 2.

The biogas production is measured using the drainage metering method, with measurements taken once daily.

The components of biogas were analyzed by gas chromatography. The analysis conditions are as follows: chromatographic column: stainless steel packed column produced by the National Chromatography Research and Analysis Center of Dalian Institute of Chemical Physics, Chinese Academy of Sciences, with TDX-01 as the filler, 2 m × 3 mn; detector: TCD; carrier gas: He, flow rate of 20 mL min−1; current, 100 mA; attenuation, 1; detection temperature, 200°C; column oven temperature, 180°C; injection temperature, 200°C.

(2) pH: The pH value is determined by the glass electrode method.

2.4.2 Determination of the content of antibiotic tetracycline

About 2 g of sample was added to a 50 mL centrifuge tube and then 10 mL of 0.1 mol/L EDTA-Mellvaine buffer; the mixture was homogenized for 1 min, sonicated for 10 min in an ice bath, and centrifuged at 10,000 rpm for 5 min at 4°C. The above steps were repeated, the supernatants from the two times were combined, and 15 mL of water-saturated n-hexane and vortex was added; centrifuge at 10,000 rpm for 5 min at 4°C; and set aside.

Solid phase extraction uses an HLB column (3 mL, 60 mg), sequentially activated with 5 mL of methanol and 5 mL of ultrapure water, accurately measures 10 mL of sample, washes with 5 mL of water, discards all liquid and dries, elutes with 6 mL of methanol, collects eluate in a 10 mL centrifuge tube, and dries. Dried with 45°C nitrogen, 0.2% formic acid water–methanol (50:50) made up to 1 mL, sonicated, passed through a 0.22-μm organic membrane filter, bottled, and tested on the machine.

3 Results and discussion

3.1 Effects of nanoparticles on the anaerobic digestion properties of OTC-containing chicken manure

3.1.1 Effects of nanoparticles on the anaerobic biogas production from OTC-containing chicken manure

Nanoparticles have a promoting effect on anaerobic gas production and gas accumulation in livestock and poultry manure containing OTC, as shown in Figure 2. In the 55-day anaerobic process, two gas production peaks appeared. The first gas production peak occurred from days 1 to 10 of the experiment, with T1–T3 reaching their peak values on day 1 without the addition of nanoparticles, at 271 ± 17, 390 ± 13, and 199 ± 15 mL, respectively. The blank group CK reached its first gas production peak on the day 3, at 166 ± 12 mL. In contrast, the T4 group with added nanoparticles reached its peak gas production on day 1, at 423 ± 21 mL, and the T5 group reached its peak on day 2, at 445 ± 31 mL. The cumulative gas production during this stage was: 2,234(T5) > 2,163(T4) > 1,445(T2) > 1,289(T3) > 1,220(T1) > 1,216(CK) mL. It can be seen that nano-Fe2O3 and nano-C60 have a promoting effect on anaerobic gas production in livestock and poultry manure containing different concentrations of OTC. In the first 10 days of the reaction, the cumulative gas production of T4 and T5 increased by 77.29 and 83.11%, respectively. After the first gas production peak (days 1–10), the system began to acidify, and gas production gradually stagnated. After adding alkaline conditions, gas production in each group resumed on day 25. The addition of OTC caused differences in the appearance of the second gas production peak in each group, with T2 and T3 reaching their peak gas production on days 32 and 33, at 360 ± 17 and 410 ± 22 mL respectively, while CK and T1 had delayed peak periods on days 36 and 38, with daily gas production of 230 ± 26 and 181 ± 37 mL, respectively. The results indicate that OTC stimulates gas production in the system, with a more pronounced effect at higher concentrations. Wang [15] found through batch experiments that 5 and 10 mg/L of OTC, respectively, increased methane production in anaerobic digestion of chicken manure by 3.38 and 7.11%, and calculated an inhibition threshold of 24.32 mg/L, which is similar to the results of this study. The stimulating effect of tetracycline antibiotics has also been observed in plants. Pomati et al. [16] found that 0.1 mg/L of tetracycline promoted the growth of duckweed, while 1 mg/L had a significant inhibitory effect on its cultivation and development.

Figure 2 Effects of nanoparticles on changes in daily biogas production rate and cumulative biogas production from anaerobic digestion of OTC-containing chicken manure.
Figure 2

Effects of nanoparticles on changes in daily biogas production rate and cumulative biogas production from anaerobic digestion of OTC-containing chicken manure.

The addition of nanoparticles T4 and T5 on day 37 reached a second peak of gas production, with volumes of 187 ± 12 and 198 ± 4 mL, respectively. The final cumulative gas production for each group was 3,712(CK), 3,993(T1), 4,344(T2), 4,447(T3), 4,671(T4), and 4,849(T5) mL. The research results show that the addition of nano-Fe2O3 and nano-C60 has a promoting effect on anaerobic gas production in livestock manure with different concentrations of OTC. Adding carbon-based nanomaterials can enhance direct interspecies electron transfer between electron-supplying bacteria and methane-producing archaea, promote the conversion of carbon dioxide to methane, shorten the lag phase required for starting biogas production, and increase methane production. Nanoparticles have excellent properties different from other common materials. The ultra-small structure of nanoparticles results in a surface effect, where the surface area, surface energy, and surface tension of nanoparticles increase sharply with decreasing particle size. This makes nanoparticles highly chemically active and easily react with other chemicals, acting as a medium to promote the gas production effect of microbial hydrolysis of substrates. At the same time, the release of metal ions from nano-Fe2O3 and nano-C60 is considered one of the most common and important ways to affect the anaerobic digestion of sludge. Fe3+ is an essential trace element for anaerobic microbial growth and metabolism and is beneficial to microorganisms within a certain range. Mu et al. [17] and others have also reached similar conclusions, suggesting that when the concentration of nano-Fe2O3 is low, the released Fe3+ can chelate with negatively charged residues in extracellular polymeric substances (EPS), reducing ion toxicity.

3.1.2 Effects of nanoparticles on the changes in pH and TVFA levels during anaerobic digestion of OTC-containing chicken manure

Nanoparticles affect the pH and TVFAs concentration changes during the anaerobic digestion process of livestock and poultry manure containing different concentrations of OTC as shown in Figures 3 and 4. VFAs are produced by hydrolysis-producing acid bacteria, among which formic acid and acetic acid are metabolic substrates for methane-producing bacteria, but excessive acid accumulation will inhibit the activity of methane-producing bacteria, reducing the gas production efficiency of the anaerobic system. As shown in Figure 4, the TVFAs concentration of each group was in a rapid increase period from day 1 to day 10, and the VFA concentration of T4 and T5 was higher than the other four groups. It can be seen that the presence of nano-Fe2O3 and nano-C60 has a promoting effect on the hydrolysis of livestock and poultry manure containing antibiotics OTC, and their ultrafine structure as a medium is conducive to the decomposition of substrates by hydrolysis-acidifying microorganisms. In the early stage of the experiment, it showed a continuous rapid increase trend, reaching the maximum peak value of 10269.2(CK), 10673.61(T1), 11056.18(T2), 10488.7(T3), 11034.5(T4) and 10,987(T5) mg/L on day 19. At this time, a large amount of free H+ was accumulated in the system due to the accumulation of TVFAs, which severely inhibited the activity of methane-producing bacteria, leading to the cessation of methane production reaction. At this time, the pH of each group continued to decrease, and on day 19, the pH of the group without added nanoparticles decreased to 5.30–5.37, while the pH of the groups with added nano-Fe2O3 and nano-C60 was lower, at 5.05 ± 0.3 and 5.03 ± 0.1 respectively. Just like the TVFAs research, the addition of nanoparticles to a certain extent promotes the hydrolysis of substrates to produce organic acids, leading to a decrease in system pH. Latif et al. [18] found that when the pH in the anaerobic digestion system was 5.5, the severe accumulation of propionic acid and butyric acid led to a 50% reduction in methane production, and the proportion of incompletely degraded organic matter would increase. When the pH value was lower than 5.5, the methane-producing community significantly decreased and shifted to a propionic acid-utilizing community. Adjusting the pH of each group to above 6.5, the activity of methane-producing bacteria recovered, and volatile acids were rapidly utilized and converted into methane. On day 34, the TVFAs concentration of CK, T1, T2, T3, T4, and T5 decreased to 5457.87, 5063.61, 4329.29, 4737.36, 5785.4, and 5876.4 mg/L, respectively, and on day 37, they rapidly increased again to 9678.20, 9411.72, 7855.33, 8263.40, 8133.3, and 8043.2 mg/L, indicating that at this time, the hydrolysis rate of difficult-to-decompose substances such as cellulose was higher than the utilization rate of methane-producing bacteria. After day 37, as the substrate decreased, the density of hydrolysis-producing acid bacteria decreased, and the TVFA concentration gradually decreased. At the end of the experiment, the pH changes of each group were small, ranging from 7.30 to 7.67.

Figure 3 Effects of nanoparticles on changes in pH value during anaerobic digestion of OTC-containing chicken manure.
Figure 3

Effects of nanoparticles on changes in pH value during anaerobic digestion of OTC-containing chicken manure.

Figure 4 Effects of nanoparticles on changes in TVFAs level during anaerobic digestion of OTC-containing chicken manure.
Figure 4

Effects of nanoparticles on changes in TVFAs level during anaerobic digestion of OTC-containing chicken manure.

3.1.3 Effects of nanoparticles on the degradation rate of OTC

The addition of nanoparticles in the anaerobic digestion process of livestock manure antibiotic OTC degradation rate is shown in Table 3. The trends of T1–T5 are the same, with OTC being rapidly degraded on days 1–5. The addition of nano-Fe2O3 and nano-C60 groups has higher degradation rates of antibiotic OTC compared to the other three groups. The degradation rate of OTC in the group without nanoparticles on day 5 is 91.34–96.29%, while the degradation rates after adding nano-Fe2O3 and nano-C60 are 96.97 and 97.08%, respectively. The concentrations then tend to stabilize, with residual amounts on day 15 being 15.25, 20.40, 56.56, 17.67, and 16.89 μg/L, and degradation rates of 98.31, 98.80, 98.29, 99.07, and 99.11%, respectively. The results show that anaerobic digestion has a significant effect on the removal of OTC. Studies have shown that under conditions without nanoparticles, adsorption is the main removal pathway for anaerobic reaction systems to remove OTC drugs [19]. Yin et al. [20] found that when the OTC addition concentration is less than 40 mg/kg TS, the anaerobic system can completely remove it. When the OTC concentration range is 40∼100 mg/kg TS, the antibiotic removal rate follows the exponential equation y = 6.6676x − 0.516. Adding 300 mg/kg TS of nano-Fe2O3 and 100 mg/kg TS of nano-C60 can further degrade the antibiotic OTC on the basis of traditional mesophilic anaerobic digestion, indicating that adding nanoparticles can promote gas production in anaerobic digestion while also having a synergistic metabolic effect on antibiotic OTC. Other studies have found that tetracycline antibiotics cannot be completely degraded in the anaerobic digestion process, and their metabolites still have similar biological toxicity to the parent compounds and are easily adsorbed to solid phases, making them more environmentally persistent. Therefore, the degradation degree of antibiotics needs to be taken seriously [21–23].

Table 3

Concentration and degradation rate of antibiotic OTC during anaerobic digestion of livestock and poultry manure

0 day (μg/L) 5 day (μg/L) Degradation rate on 5 day (%) 15 day (μg/L) Degradation rate on 15 day (%)
T1 700 65.75 92.69 15.25 98.31
T2 1,900 63.13 96.29 20.40 98.80
T3 3,300 285.83 91.34 56.56 98.29
T4 1,900 57.66 96.97 17.67 99.07
T5 1,900 55.43 97.08 16.89 99.11

3.1.4 Effects of OTC on the activities of different enzymes

Anaerobic digestion produces biogas, which is the result of enzymatic reactions gradually carried out under the action of different functional microbial communities. The activity of dehydrogenase is an important indicator of microbial status. The changes in dehydrogenase activity during anaerobic digestion with different concentrations of OTC are shown in Figure 5. The concentration of dehydrogenase showed a trend of first increasing and then decreasing throughout the entire experimental period. The dehydrogenase activity in the antibiotic group was higher than that in the control group. The addition of 300 mg/kg TS of nano-Fe2O3 and 100 mg/kg TS of nano-C60 had the highest dehydrogenase activity. The average dehydrogenase activity of each group was 127.02(T5) > 124.74(T4) > 114.38(T2) > 114.98(T3) > 113.87(T1) > 90.88(CK) IU/L, indicating that the addition of nanoparticles stimulates microbial activity.

Figure 5 Changes in dehydrogenase activity at different OTC concentrations.
Figure 5

Changes in dehydrogenase activity at different OTC concentrations.

The changes in anaerobic digestion enzyme activity of different concentrations of OTC are shown in Figure 6. The addition of antibiotics and nano-particle groups showed a trend of first increase and then decrease in amylase activity, while the control group showed no significant changes. On the first day, there was no significant difference in amylase activity among the groups, all ranging 291.37–321.21 IU/L. On the fifth day, the amylase activity in the control group decreased to 242.47 IU/L, while T1–T3 were stimulated by OTC, with significantly increased amylase activity of 321.61, 313.84, and 366.44 IU/L, respectively, and the stimulating effect continued until the end of the experiment, with T4 and T5 groups at 324.3 and 332.5 IU/L, respectively. T3 reached its maximum value of 429.86 IU/L on day 15, while T1 and T2 reached their maximum values of 406.56 and 461.16 IU/L on day 30. The mean enzyme activity of each group was 384.08(T5) > 381.48(T2) > 378.12(T4) > 365.35(T3) > 364.79(T1) > 268.26(CK) IU/L. OTC has a stimulating effect on amylase activity, with the best promoting effect at a concentration of 50 mg/kg TS, and the addition of 100 mg/kg TS nano-C60 has a better stimulating effect on amylase activity.

Figure 6 Changes in activities of hydrolytic enzymes at different OTC concentrations.
Figure 6

Changes in activities of hydrolytic enzymes at different OTC concentrations.

Cellulose can be hydrolyzed into cellobiose by cellulase, and then further broken down into glucose. The change in cellulase activity can reflect the degradation of carbon substances during the fermentation process [24]. Chicken manure has a high cellulose content, with a longer hydrolysis time compared to starch and protein. The cellulase activity level in the system is significantly higher than other hydrolytic enzymes. As shown in Figure 5, on the first day, the cellulase activity in the antibiotic group was significantly enhanced. On the fifth day, it was the peak period of the first gas production, with enzyme activity levels of 131.77(CK), 251.21(T1), 292.50(T2), 198.96(T3), 273.6(T4), and 265.9(T5) IU/mL. The addition of 300 mg/kg TS of nano-Fe2O3 and 100 mg/kg TS of nano-C60 resulted in the highest cellulase activity. On day 30, each group was in the peak period of the second gas production, with enzyme activity data of 158.46(CK), 263.32(T1), 259.13(T1), 248.84(T3), 274.6(T4), and 289.9(T5) IU/mL. The difference in enzyme activity between the groups was small, but all were significantly higher than the control. On day 15, group T2 reached the maximum enzyme activity of 358.79 IU/mL. As cellulose was utilized, the density of cellulose-hydrolyzing bacteria decreased, leading to a significant decrease in enzyme activity. The average enzyme activity values were 275.43(T2) > 259.34(T5) > 253.41(T4) > 243.84(T3) > 242.26(T1) > 143.98(CK), indicating that tilmicosin stimulates cellulase activity, with the best promoting effect at a concentration of 50 mg/kg TS.

Determining the activities of protease and urease is of great significance for understanding the decomposition and metabolism of nitrogenous organic matter in anaerobic digestion slurry and the changes in ammonia nitrogen concentration. Protein degradation is divided into two stages: extracellular and intracellular. Proteins are broken down into proteose and peptides under the catalysis of protease and eventually converted into amino acids that can be utilized by methane-producing bacteria. The average activities of protease are 227.56(T4) > 227.21(T2) > 224.21(T5) > 219.48(T3) > 204.17(T1) > 174.52(CK) IU/L. The protease activity in the antibiotic group is higher than in the control group, indicating that tetracycline stimulates protease. Among them, the protease activity level of T2 is the highest, and the addition of nanoparticles has no significant effect on protease activity. Urease activity data shows that the antibiotic group generally shows a trend of first increase and then decrease, while the control group shows little change over time. Throughout the experiment, the urease activity of CK has been maintained at 40 IU/mL. On the first day, the antibiotic group was significantly higher than the control group, and then, the difference in urease activity between T1–T3 and CK continued to increase until day 15. T1, T2, and T5 reached their maximum values at 15 days, which were 67.04, 73.06, and 75.05 IU/mL, respectively. Groups T3 and T4 reached their maximum values at 30 days, which were 68.20 and 69.53 IU/mL. The average activities of the enzymes in each group are 69.21(T5) > 64.79(T4) > 62.83(T2) > 54.84(T3) > 52.83(T1) > 38.78(CK) IU/mL. The addition of nanoparticles T4 and T5 has a promoting effect on urease activity. Pan et al. [25] found that 60 and 100 mg/kg tetracycline activates urease activity in the early stage of anaerobic fermentation and has a certain promoting effect on methane production. Gas production was significantly inhibited in the later stages of the experiment. They speculated that in the early stages of the experiment, urease may use the amide bond of tetracycline as a substrate and induce an increase in urease activity through substrate induction.

3.2 Nanoparticles on OTC-resistant genes in livestock and poultry manure

The anaerobic digestion system itself has a certain tolerance, and the self-regulation ability during operation is also a guarantee to cope with toxic environments. Yu et al. [26] found through a 180-h OTC waste sludge batch mesophilic anaerobic digestion experiment that 5 mg/L tetracycline did not have a negative impact on the anaerobic digestion system, while 50 mg/L tetracycline would acutely inhibit methane production activity in the system, with inhibition relieved at 141 hours and methane production reduced by 23.75%. Wei et al. [27] found through OTC pig manure mesophilic batch anaerobic digestion experiments that 10–1,000 mg/kg tetracycline could significantly increase biogas production, with 50 mg/kg showing the best promoting effect. When the OTC addition concentration was 2,000 mg/kg, it would severely inhibit microbial activity. The inoculum used by Wei was domesticated with antibiotics before use, showing a certain degree of resistance. Although the antibiotic content in the chicken manure and activated sludge used in this study was below the detection limit, the metagenomic sequencing results in Figure 7 showed that there were various types of ARGs in groups T2, T4, and T5, with the multidrug resistance gene (ARGs)-Multidrug accounting for a high proportion of 33.54, 35.63, and 37.43%, respectively, indicating a certain resistance of the anaerobic digestion system to tetracycline. The proportions of the other four types of MLS, tetracycline, glycopeptide, and peptide were between 8.18% and 12.98%. It can be seen that the addition of 300 mg/kg TS of nano-Fe2O3 and 100 mg/kg TS of nano-C60 can increase the proportion of multidrug resistance genes, while not reducing the proportions of MLS, tetracycline, glycopeptide, and peptide resistance genes, to a certain extent enhancing the inhibitory effect of the anaerobic digestion system on tetracycline. The use of antibiotics in agricultural breeding processes can lead to the presence of resistant bacteria and resistance genes in the intestines of livestock and poultry, which are then excreted with feces. Wastewater treatment systems serve as both a “sink” and a “source” of antibiotics, with activated sludge becoming the main carrier of ARGs. The higher biological density in anaerobic reactors may promote gene transfer, with resistant bacteria gaining an advantage under the selection pressure of antibiotics [28]. In addition, research [29] has shown that when microorganisms are exposed to toxic substances, EPS can release substrates to bind with toxic substances and form a protective barrier, which may be an important reason why anaerobic systems can resist toxic substances.

Figure 7 (a), (b) and (c) represent pie charts of ARGs for chicken manure and sludge in T2, T4 and T5 groups, respectively.
Figure 7

(a), (b) and (c) represent pie charts of ARGs for chicken manure and sludge in T2, T4 and T5 groups, respectively.

4 Conclusions

The addition of nano-Fe2O3 and nano-C60 has a promoting effect on the anaerobic gas production of livestock and poultry manure containing different concentrations of OTC. The nano-particle group T4 reached peak gas production on the first day, with 423 ± 21 mL, while group T5 reached peak gas production on the second day, with 445 ± 31 mL. The cumulative gas production from day 1 to day 10 was 2,234(T5) > 2,163(T4) > 1,445(T2) > 1,289(T3) > 1,220(T1) > 1,216(CK) mL. The cumulative gas production of T4 and T5 increased by 77.29 and 83.11%, respectively. The cumulative gas production of each group was 3,712(CK), 3,993(T1), 4,344(T2), 4,447(T3), 4,671(T4), and 4,849(T5) mL.

The trend of changes in the degradation rate of antibiotic OTC during anaerobic digestion of livestock and poultry manure in T1–T5 is the same. The degradation rate of OTC without the addition of nanoparticles on the fifth day is 91.34–96.29%. The degradation rates after adding nano-Fe2O3 and nano-C60 are 96.97 and 97.08%, respectively. The concentrations then tend to stabilize, with residual amounts on day 15 being 15.25, 20.40, 56.56, 17.67, and 16.89 μg/L, and degradation rates being 98.31, 98.80, 98.29, 99.07, and 99.11%, respectively.

Over-the-counter antibiotics can increase the activity of dehydrogenase, amylase, protease, and urease. Adding nanoparticles can increase the activity of dehydrogenase and amylase, with no significant impact on cellulase, urease, and protease activity.

There are multiple types of ARGs in groups T2, T4, and T5, with the proportion of multidrug resistance genes (ARGs)-Multidrug being relatively high at 33.54, 35.63, and 37.43%, respectively. The proportions of the other four types of ARGs, MLS, tetracycline, glycopeptide, and peptide range from 8.18–12.98%.

  1. Funding information: This work was funded by the National Natural Science Foundation of China (52206255), the Youth Science and Technology Talent Lift Program of Gansu Province (GXH20220530-14), the Youth Science Foundation Project of Lanzhou Jiaotong University (2020018, 2020011), and Gansu Province Youth Science Fund project (No. 22JR11RA148).

  2. Author contributions: Xiaofei Zhen: writing – original draft, conceptualization; Ruonan Jiao: writing – original draft, conceptualization; Han Zhan: writing – review and editing; Wenbing Wu: investigation; Ke Li: investigation; Lei Feng: supervision; Tie Du: validation.

  3. Conflict of interest: The authors state no conflict of interest.

  4. Data availability statement: The data sets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

  5. Ethical approval: The conducted research is not related to either human or animal use.

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Received: 2024-04-22
Revised: 2024-06-04
Accepted: 2024-06-14
Published Online: 2024-07-12

© 2024 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/chem-2024-0060
Keywords for this article
nanoparticles; oxytetracycline; anaerobic digestion; antibiotics; resistance genes
Creative Commons
BY 4.0

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  9. Two triazole-based coordination polymers: Synthesis and crystal structure characterization
  10. Phytochemical and physicochemical studies of different apple varieties grown in Morocco
  11. Synthesis of multi-template molecularly imprinted polymers (MT-MIPs) for isolating ethyl para-methoxycinnamate and ethyl cinnamate from Kaempferia galanga L., extract with methacrylic acid as functional monomer
  12. Nutraceutical potential of Mesembryanthemum forsskaolii Hochst. ex Bioss.: Insights into its nutritional composition, phytochemical contents, and antioxidant activity
  13. Evaluation of influence of Butea monosperma floral extract on inflammatory biomarkers
  14. Cannabis sativa L. essential oil: Chemical composition, anti-oxidant, anti-microbial properties, and acute toxicity: In vitro, in vivo, and in silico study
  15. The effect of gamma radiation on 5-hydroxymethylfurfural conversion in water and dimethyl sulfoxide
  16. Hollow mushroom nanomaterials for potentiometric sensing of Pb2+ ions in water via the intercalation of iodide ions into the polypyrrole matrix
  17. Determination of essential oil and chemical composition of St. John’s Wort
  18. Computational design and in vitro assay of lantadene-based novel inhibitors of NS3 protease of dengue virus
  19. Anti-parasitic activity and computational studies on a novel labdane diterpene from the roots of Vachellia nilotica
  20. Microbial dynamics and dehydrogenase activity in tomato (Lycopersicon esculentum Mill.) rhizospheres: Impacts on growth and soil health across different soil types
  21. Correlation between in vitro anti-urease activity and in silico molecular modeling approach of novel imidazopyridine–oxadiazole hybrids derivatives
  22. Spatial mapping of indoor air quality in a light metro system using the geographic information system method
  23. Iron indices and hemogram in renal anemia and the improvement with Tribulus terrestris green-formulated silver nanoparticles applied on rat model
  24. Integrated track of nano-informatics coupling with the enrichment concept in developing a novel nanoparticle targeting ERK protein in Naegleria fowleri
  25. Cytotoxic and phytochemical screening of Solanum lycopersicum–Daucus carota hydro-ethanolic extract and in silico evaluation of its lycopene content as anticancer agent
  26. Protective activities of silver nanoparticles containing Panax japonicus on apoptotic, inflammatory, and oxidative alterations in isoproterenol-induced cardiotoxicity
  27. pH-based colorimetric detection of monofunctional aldehydes in liquid and gas phases
  28. Investigating the effect of resveratrol on apoptosis and regulation of gene expression of Caco-2 cells: Unravelling potential implications for colorectal cancer treatment
  29. Metformin inhibits knee osteoarthritis induced by type 2 diabetes mellitus in rats: S100A8/9 and S100A12 as players and therapeutic targets
  30. Effect of silver nanoparticles formulated by Silybum marianum on menopausal urinary incontinence in ovariectomized rats
  31. Synthesis of new analogs of N-substituted(benzoylamino)-1,2,3,6-tetrahydropyridines
  32. Response of yield and quality of Japonica rice to different gradients of moisture deficit at grain-filling stage in cold regions
  33. Preparation of an inclusion complex of nickel-based β-cyclodextrin: Characterization and accelerating the osteoarthritis articular cartilage repair
  34. Empagliflozin-loaded nanomicelles responsive to reactive oxygen species for renal ischemia/reperfusion injury protection
  35. Preparation and pharmacodynamic evaluation of sodium aescinate solid lipid nanoparticles
  36. Assessment of potentially toxic elements and health risks of agricultural soil in Southwest Riyadh, Saudi Arabia
  37. Theoretical investigation of hydrogen-rich fuel production through ammonia decomposition
  38. Biosynthesis and screening of cobalt nanoparticles using citrus species for antimicrobial activity
  39. Investigating the interplay of genetic variations, MCP-1 polymorphism, and docking with phytochemical inhibitors for combatting dengue virus pathogenicity through in silico analysis
  40. Ultrasound induced biosynthesis of silver nanoparticles embedded into chitosan polymers: Investigation of its anti-cutaneous squamous cell carcinoma effects
  41. Copper oxide nanoparticles-mediated Heliotropium bacciferum leaf extract: Antifungal activity and molecular docking assays against strawberry pathogens
  42. Sprouted wheat flour for improving physical, chemical, rheological, microbial load, and quality properties of fino bread
  43. Comparative toxicity assessment of fisetin-aided artificial intelligence-assisted drug design targeting epibulbar dermoid through phytochemicals
  44. Acute toxicity and anti-inflammatory activity of bis-thiourea derivatives
  45. Anti-diabetic activity-guided isolation of α-amylase and α-glucosidase inhibitory terpenes from Capsella bursa-pastoris Linn.
  46. GC–MS analysis of Lactobacillus plantarum YW11 metabolites and its computational analysis on familial pulmonary fibrosis hub genes
  47. Green formulation of copper nanoparticles by Pistacia khinjuk leaf aqueous extract: Introducing a novel chemotherapeutic drug for the treatment of prostate cancer
  48. Improved photocatalytic properties of WO3 nanoparticles for Malachite green dye degradation under visible light irradiation: An effect of La doping
  49. One-pot synthesis of a network of Mn2O3–MnO2–poly(m-methylaniline) composite nanorods on a polypyrrole film presents a promising and efficient optoelectronic and solar cell device
  50. Groundwater quality and health risk assessment of nitrate and fluoride in Al Qaseem area, Saudi Arabia
  51. A comparative study of the antifungal efficacy and phytochemical composition of date palm leaflet extracts
  52. Processing of alcohol pomelo beverage (Citrus grandis (L.) Osbeck) using saccharomyces yeast: Optimization, physicochemical quality, and sensory characteristics
  53. Specialized compounds of four Cameroonian spices: Isolation, characterization, and in silico evaluation as prospective SARS-CoV-2 inhibitors
  54. Identification of a novel drug target in Porphyromonas gingivalis by a computational genome analysis approach
  55. Physico-chemical properties and durability of a fly-ash-based geopolymer
  56. FMS-like tyrosine kinase 3 inhibitory potentials of some phytochemicals from anti-leukemic plants using computational chemical methodologies
  57. Wild Thymus zygis L. ssp. gracilis and Eucalyptus camaldulensis Dehnh.: Chemical composition, antioxidant and antibacterial activities of essential oils
  58. 3D-QSAR, molecular docking, ADMET, simulation dynamic, and retrosynthesis studies on new styrylquinolines derivatives against breast cancer
  59. Deciphering the influenza neuraminidase inhibitory potential of naturally occurring biflavonoids: An in silico approach
  60. Determination of heavy elements in agricultural regions, Saudi Arabia
  61. Synthesis and characterization of antioxidant-enriched Moringa oil-based edible oleogel
  62. Ameliorative effects of thistle and thyme honeys on cyclophosphamide-induced toxicity in mice
  63. Study of phytochemical compound and antipyretic activity of Chenopodium ambrosioides L. fractions
  64. Investigating the adsorption mechanism of zinc chloride-modified porous carbon for sulfadiazine removal from water
  65. Performance repair of building materials using alumina and silica composite nanomaterials with electrodynamic properties
  66. Effects of nanoparticles on the activity and resistance genes of anaerobic digestion enzymes in livestock and poultry manure containing the antibiotic tetracycline
  67. Effect of copper nanoparticles green-synthesized using Ocimum basilicum against Pseudomonas aeruginosa in mice lung infection model
  68. Cardioprotective effects of nanoparticles green formulated by Spinacia oleracea extract on isoproterenol-induced myocardial infarction in mice by the determination of PPAR-γ/NF-κB pathway
  69. Anti-OTC antibody-conjugated fluorescent magnetic/silica and fluorescent hybrid silica nanoparticles for oxytetracycline detection
  70. Curcumin conjugated zinc nanoparticles for the treatment of myocardial infarction
  71. Identification and in silico screening of natural phloroglucinols as potential PI3Kα inhibitors: A computational approach for drug discovery
  72. Exploring the phytochemical profile and antioxidant evaluation: Molecular docking and ADMET analysis of main compounds from three Solanum species in Saudi Arabia
  73. Unveiling the molecular composition and biological properties of essential oil derived from the leaves of wild Mentha aquatica L.: A comprehensive in vitro and in silico exploration
  74. Analysis of bioactive compounds present in Boerhavia elegans seeds by GC-MS
  75. Homology modeling and molecular docking study of corticotrophin-releasing hormone: An approach to treat stress-related diseases
  76. LncRNA MIR17HG alleviates heart failure via targeting MIR17HG/miR-153-3p/SIRT1 axis in in vitro model
  77. Development and validation of a stability indicating UPLC-DAD method coupled with MS-TQD for ramipril and thymoquinone in bioactive SNEDDS with in silico toxicity analysis of ramipril degradation products
  78. Biosynthesis of Ag/Cu nanocomposite mediated by Curcuma longa: Evaluation of its antibacterial properties against oral pathogens
  79. Development of AMBER-compliant transferable force field parameters for polytetrafluoroethylene
  80. Treatment of gestational diabetes by Acroptilon repens leaf aqueous extract green-formulated iron nanoparticles in rats
  81. Development and characterization of new ecological adsorbents based on cardoon wastes: Application to brilliant green adsorption
  82. A fast, sensitive, greener, and stability-indicating HPLC method for the standardization and quantitative determination of chlorhexidine acetate in commercial products
  83. Assessment of Se, As, Cd, Cr, Hg, and Pb content status in Ankang tea plantations of China
  84. Effect of transition metal chloride (ZnCl2) on low-temperature pyrolysis of high ash bituminous coal
  85. Evaluating polyphenol and ascorbic acid contents, tannin removal ability, and physical properties during hydrolysis and convective hot-air drying of cashew apple powder
  86. Development and characterization of functional low-fat frozen dairy dessert enhanced with dried lemongrass powder
  87. Scrutinizing the effect of additive and synergistic antibiotics against carbapenem-resistant Pseudomonas aeruginosa
  88. Preparation, characterization, and determination of the therapeutic effects of copper nanoparticles green-formulated by Pistacia atlantica in diabetes-induced cardiac dysfunction in rat
  89. Antioxidant and antidiabetic potentials of methoxy-substituted Schiff bases using in vitro, in vivo, and molecular simulation approaches
  90. Anti-melanoma cancer activity and chemical profile of the essential oil of Seseli yunnanense Franch
  91. Molecular docking analysis of subtilisin-like alkaline serine protease (SLASP) and laccase with natural biopolymers
  92. Overcoming methicillin resistance by methicillin-resistant Staphylococcus aureus: Computational evaluation of napthyridine and oxadiazoles compounds for potential dual inhibition of PBP-2a and FemA proteins
  93. Exploring novel antitubercular agents: Innovative design of 2,3-diaryl-quinoxalines targeting DprE1 for effective tuberculosis treatment
  94. Drimia maritima flowers as a source of biologically potent components: Optimization of bioactive compound extractions, isolation, UPLC–ESI–MS/MS, and pharmacological properties
  95. Estimating molecular properties, drug-likeness, cardiotoxic risk, liability profile, and molecular docking study to characterize binding process of key phyto-compounds against serotonin 5-HT2A receptor
  96. Fabrication of β-cyclodextrin-based microgels for enhancing solubility of Terbinafine: An in-vitro and in-vivo toxicological evaluation
  97. Phyto-mediated synthesis of ZnO nanoparticles and their sunlight-driven photocatalytic degradation of cationic and anionic dyes
  98. Monosodium glutamate induces hypothalamic–pituitary–adrenal axis hyperactivation, glucocorticoid receptors down-regulation, and systemic inflammatory response in young male rats: Impact on miR-155 and miR-218
  99. Quality control analyses of selected honey samples from Serbia based on their mineral and flavonoid profiles, and the invertase activity
  100. Eco-friendly synthesis of silver nanoparticles using Phyllanthus niruri leaf extract: Assessment of antimicrobial activity, effectiveness on tropical neglected mosquito vector control, and biocompatibility using a fibroblast cell line model
  101. Green synthesis of silver nanoparticles containing Cichorium intybus to treat the sepsis-induced DNA damage in the liver of Wistar albino rats
  102. Quality changes of durian pulp (Durio ziberhinus Murr.) in cold storage
  103. Study on recrystallization process of nitroguanidine by directly adding cold water to control temperature
  104. Determination of heavy metals and health risk assessment in drinking water in Bukayriyah City, Saudi Arabia
  105. Larvicidal properties of essential oils of three Artemisia species against the chemically insecticide-resistant Nile fever vector Culex pipiens (L.) (Diptera: Culicidae): In vitro and in silico studies
  106. Design, synthesis, characterization, and theoretical calculations, along with in silico and in vitro antimicrobial proprieties of new isoxazole-amide conjugates
  107. The impact of drying and extraction methods on total lipid, fatty acid profile, and cytotoxicity of Tenebrio molitor larvae
  108. A zinc oxide–tin oxide–nerolidol hybrid nanomaterial: Efficacy against esophageal squamous cell carcinoma
  109. Research on technological process for production of muskmelon juice (Cucumis melo L.)
  110. Physicochemical components, antioxidant activity, and predictive models for quality of soursop tea (Annona muricata L.) during heat pump drying
  111. Characterization and application of Fe1−xCoxFe2O4 nanoparticles in Direct Red 79 adsorption
  112. Torilis arvensis ethanolic extract: Phytochemical analysis, antifungal efficacy, and cytotoxicity properties
  113. Magnetite–poly-1H pyrrole dendritic nanocomposite seeded on poly-1H pyrrole: A promising photocathode for green hydrogen generation from sanitation water without using external sacrificing agent
  114. HPLC and GC–MS analyses of phytochemical compounds in Haloxylon salicornicum extract: Antibacterial and antifungal activity assessment of phytopathogens
  115. Efficient and stable to coking catalysts of ethanol steam reforming comprised of Ni + Ru loaded on MgAl2O4 + LnFe0.7Ni0.3O3 (Ln = La, Pr) nanocomposites prepared via cost-effective procedure with Pluronic P123 copolymer
  116. Nitrogen and boron co-doped carbon dots probe for selectively detecting Hg2+ in water samples and the detection mechanism
  117. Heavy metals in road dust from typical old industrial areas of Wuhan: Seasonal distribution and bioaccessibility-based health risk assessment
  118. Phytochemical profiling and bioactivity evaluation of CBD- and THC-enriched Cannabis sativa extracts: In vitro and in silico investigation of antioxidant and anti-inflammatory effects
  119. Investigating dye adsorption: The role of surface-modified montmorillonite nanoclay in kinetics, isotherms, and thermodynamics
  120. Antimicrobial activity, induction of ROS generation in HepG2 liver cancer cells, and chemical composition of Pterospermum heterophyllum
  121. Study on the performance of nanoparticle-modified PVDF membrane in delaying membrane aging
  122. Impact of cholesterol in encapsulated vitamin E acetate within cocoliposomes
  123. Review Articles
  124. Structural aspects of Pt(η3-X1N1X2)(PL) (X1,2 = O, C, or Se) and Pt(η3-N1N2X1)(PL) (X1 = C, S, or Se) derivatives
  125. Biosurfactants in biocorrosion and corrosion mitigation of metals: An overview
  126. Stimulus-responsive MOF–hydrogel composites: Classification, preparation, characterization, and their advancement in medical treatments
  127. Electrochemical dissolution of titanium under alternating current polarization to obtain its dioxide
  128. Special Issue on Recent Trends in Green Chemistry
  129. Phytochemical screening and antioxidant activity of Vitex agnus-castus L.
  130. Phytochemical study, antioxidant activity, and dermoprotective activity of Chenopodium ambrosioides (L.)
  131. Exploitation of mangliculous marine fungi, Amarenographium solium, for the green synthesis of silver nanoparticles and their activity against multiple drug-resistant bacteria
  132. Study of the phytotoxicity of margines on Pistia stratiotes L.
  133. Special Issue on Advanced Nanomaterials for Energy, Environmental and Biological Applications - Part III
  134. Impact of biogenic zinc oxide nanoparticles on growth, development, and antioxidant system of high protein content crop (Lablab purpureus L.) sweet
  135. Green synthesis, characterization, and application of iron and molybdenum nanoparticles and their composites for enhancing the growth of Solanum lycopersicum
  136. Green synthesis of silver nanoparticles from Olea europaea L. extracted polysaccharides, characterization, and its assessment as an antimicrobial agent against multiple pathogenic microbes
  137. Photocatalytic treatment of organic dyes using metal oxides and nanocomposites: A quantitative study
  138. Antifungal, antioxidant, and photocatalytic activities of greenly synthesized iron oxide nanoparticles
  139. Special Issue on Phytochemical and Pharmacological Scrutinization of Medicinal Plants
  140. Hepatoprotective effects of safranal on acetaminophen-induced hepatotoxicity in rats
  141. Chemical composition and biological properties of Thymus capitatus plants from Algerian high plains: A comparative and analytical study
  142. Chemical composition and bioactivities of the methanol root extracts of Saussurea costus
  143. In vivo protective effects of vitamin C against cyto-genotoxicity induced by Dysphania ambrosioides aqueous extract
  144. Insights about the deleterious impact of a carbamate pesticide on some metabolic immune and antioxidant functions and a focus on the protective ability of a Saharan shrub and its anti-edematous property
  145. A comprehensive review uncovering the anticancerous potential of genkwanin (plant-derived compound) in several human carcinomas
  146. A study to investigate the anticancer potential of carvacrol via targeting Notch signaling in breast cancer
  147. Assessment of anti-diabetic properties of Ziziphus oenopolia (L.) wild edible fruit extract: In vitro and in silico investigations through molecular docking analysis
  148. Optimization of polyphenol extraction, phenolic profile by LC-ESI-MS/MS, antioxidant, anti-enzymatic, and cytotoxic activities of Physalis acutifolia
  149. Phytochemical screening, antioxidant properties, and photo-protective activities of Salvia balansae de Noé ex Coss
  150. Antihyperglycemic, antiglycation, anti-hypercholesteremic, and toxicity evaluation with gas chromatography mass spectrometry profiling for Aloe armatissima leaves
  151. Phyto-fabrication and characterization of gold nanoparticles by using Timur (Zanthoxylum armatum DC) and their effect on wound healing
  152. Does Erodium trifolium (Cav.) Guitt exhibit medicinal properties? Response elements from phytochemical profiling, enzyme-inhibiting, and antioxidant and antimicrobial activities
  153. Integrative in silico evaluation of the antiviral potential of terpenoids and its metal complexes derived from Homalomena aromatica based on main protease of SARS-CoV-2
  154. 6-Methoxyflavone improves anxiety, depression, and memory by increasing monoamines in mice brain: HPLC analysis and in silico studies
  155. Simultaneous extraction and quantification of hydrophilic and lipophilic antioxidants in Solanum lycopersicum L. varieties marketed in Saudi Arabia
  156. Biological evaluation of CH3OH and C2H5OH of Berberis vulgaris for in vivo antileishmanial potential against Leishmania tropica in murine models
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