|Year : 2022 | Volume
| Issue : 3 | Page : 353-359
Inhibitory effect of Basella alba-Mediated zinc oxide nanoparticles against the infection-causing bacteria
Aranganathan Agila1, Gnanasekar Dayana Jeyaleela2, Joseph Devaraj Rosaline Vimala1, Moses Stella Bharathy1, Sagaya Adaikalaraj Margrat Sheela1
1 Department of Chemistry, Holy Cross College, Affiliated to Bharathidasan University, Tiruchirappalli, Tamil Nadu, India
2 Department of Chemistry, Shrimati Indira Gandhi College, Affiliated to Bharathidasan University, Tiruchirappalli, Tamil Nadu, India
|Date of Submission||05-Jun-2022|
|Date of Decision||25-Jul-2022|
|Date of Acceptance||01-Aug-2022|
|Date of Web Publication||17-Sep-2022|
PG & Research Department of Chemistry, Holy Cross College (Autonomous), Affiliated to Bharathidasan University, Tiruchirappalli, Tamil Nadu
Source of Support: None, Conflict of Interest: None
Background: Naturally occurring biomolecules from the plant extract have been identified to play an active role in the formation of any nanoparticles. Methods: This work aimed to synthesize the nano-sized zinc oxide material (zinc oxide nanoparticles [ZnONPs]) using the 70% ethanolic leaf extract of Basella alba by precipitation method and also studied the antibacterial activity of green-synthesized ZnONPs on infection-causing five bacteria. Synthesized nanomaterials were characterized by the aid of ultraviolet (UV)-visible, Fourier transform infrared spectroscopy, scanning electron microscope, X-ray diffraction, and Energy Dispersive X-ray Analysis (EDAX). Results: Qualitative analysis and UV results of extract reveal the occurrence of some medicinally important phytomolecules such as flavonoids, terpenoids, phenolic acids, and ascorbic acid. The spherical nature of ZnONPs was observed with an average crystalline size of 28.64 nm. EDAX analysis revealed the elemental compositions in the B. alba-mediated ZnONPs (BA-ZnONPs) which showed zinc in 70.04% and oxygen in 29.96%. BA-ZnONPs were tested against the bacteria (an infection causing) such as Pseudomonas aeruginosa, Escherichia coli, Enterobacter aerogenes, Staphylococcus aureus, and Proteus vulgaris, which results that, against all the pathogens, ZnONPs showed noticeable inhibition effects compared with zinc acetate and B. alba extract. Especially against the E. coli, ZnONPs performed well with inhibitory effect and least on S. aureus. Conclusion: Antibacterial activities of BA-ZnONPs were studied which can act as the new antimicrobial-resistant agents.
Keywords: Antibacterial activity on infection-causing bacteria, Basella alba, characterization, green synthesis
|How to cite this article:|
Agila A, Dayana Jeyaleela G, Vimala JD, Bharathy MS, Sheela SA. Inhibitory effect of Basella alba-Mediated zinc oxide nanoparticles against the infection-causing bacteria. Biomed Biotechnol Res J 2022;6:353-9
|How to cite this URL:|
Agila A, Dayana Jeyaleela G, Vimala JD, Bharathy MS, Sheela SA. Inhibitory effect of Basella alba-Mediated zinc oxide nanoparticles against the infection-causing bacteria. Biomed Biotechnol Res J [serial online] 2022 [cited 2022 Oct 5];6:353-9. Available from: https://www.bmbtrj.org/text.asp?2022/6/3/353/356145
| Introduction|| |
Numerous Gram-negative and Gram-positive bacteria are identified that can affect any internal and external organs or part(s) of the human body. Bacterial infections are the main causes of chronic infections and mortality. Any infectious diseases will be caused by the bacteria when they get into the human body, and they produce some chemicals called toxins which can be damaged or destroyed the tissues. Bacterial infections occur in various types which including cystitis (urinary tract), cellulitis (skin), pharyngitis (respiration system), tonsillitis (respiratory tract), toxemia (bloodstream), osteomyelitis (bone), pneumonia (clinical), septic arthritis (joint), and necrotizing and pyoderma (soft tissue).,
Nanotechnology, abbreviated as “nanotech,” is the intriguingly emerging recent technology that involves the manipulation of the bulk materials into nano-sized materials and is ranging from 1 to 100 nm. It has been practiced extensively in curative applications, which include biological applications and drug delivery to improve and develop therapeutic applications for numerous diseases. Nanoscience covers all the science domains known to humankind including physics, chemistry, human biology, genetics, energy sciences, food industry, electronics, aviation, medicine, or any other field of scientific research. Nanomaterials/particles have been long known for their excellent antimicrobial behavior against most Gram-positive and Gram-negative bacterial pathogens. Even the metal oxide nanoparticles also serve as tremendous antimicrobial agents because they have a large surface area. Nowadays, nanomaterial-based drugs were potentially used as alternate antibiotics for all infectious diseases (bacterial and fungal)., Zinc oxide nanoparticles (ZnONPs) also showed excellent therapeutic effects, especially against the infectious disease it used to control as well as treat infections. In addition, zinc oxide (ZnO) particles revealed wound-healing properties, antioxidant effects, and anticancer properties.
In-plant extract-mediated nanomaterials, i.e., excess of phytocompounds, were involved in the synthesis which can affect the shape, size, and property of the resulted ZnO nanomaterials. The utilization of medicinal plant extracts may reduce the toxicity of the nanomaterials and thus enhance the medicinal applications in pharmacological fields. The main advantage of practicing plants is the unnecessary usage of capping and stabilizing agents.,, The plant extracts act as all the three reagent's roles because of the presence of many phytocompounds present in the single extract. The plant chosen for the study is Basella alba in which the leaves are the edible perennial vine and belong to the Basellaceae family. It is mostly used as a leaf vegetable and is widely found in tropical Asia and Africa. Commonly, it is known as vine spinach, Malabar spinach, and Ceylon spinach and is very cheap efficacy and easily accessible. It possesses lots of medicinal values such as antidiabetic, antimicrobial, antioxidant, antiviral, and anticancer. The red variety of B. alba was historically used for red dye preparation in China.,,,,
From the above discussion, ZnO material possesses satisfactory antimicrobial effects. Hence, the present study aimed to biosynthesize and characterization of B. alba extract mediated Zinc oxide nanoparticles and also aimed to screen its antibacterial potential against the infection-causing bacteria.
| Methods|| |
Preparation of 70% ethanolic extract of Basella alba leaves
About 500-g disease-free leaves of B. alba were chopped and ground and then transferred into the 500-ml beaker; to that, 70% ethanol was added and mixed well. Maintain the solvent level in the beaker at 2 cm above the plant material, then kept for 3 days for the complete extraction of all the phytoconstituents in the plant leaves. The final extract was first filtered by using the ordinary filter paper and then followed by Whatman filter paper. The final liquid filtrate was kept in the refrigerator for further work. The different qualitative chemical tests can be performed to establish or screen the phytochemical profile of 70% ethanolic extract of B. alba. It is an essential test for detecting bioactive compounds (plant products) present in the different extracts of plants. The standard screening tests and procedures were carried out which are performed on the extract to detect various phytocompounds present in them.
Synthesis and characterization of Basella alba extract-mediated zinc oxide nanoparticles
50 ml of 0.1 M concentrated zinc acetate solution was prepared in a 250-ml beaker and kept on the magnetic stirrer. To this, 10 ml of the filtered ethanolic extract of B. alba was slowly added dropwise at every 30-min time interval. Then, the temperature was set to 60°C; after the complete addition of 50 ml of ethanolic plant extract, the whole mixture was left for 24 h at room temperature for the nanoparticle growth. ZnONPs were deposited in the beaker bottom, and it was collected by centrifuging at 6000 rpm. To achieve the maximum purity of ZnONPs, the collected ZnONPs were washed with Double distilled water, followed by ethanol. Solvent-free ZnONPs were calcinated for the complete conversion of Zn(OH)2 into ZnO. Synthesized ZnONPs were confirmed with ultraviolet (UV)-visible spectroscopy, Fourier transform infrared spectroscopy (FT-IR) spectral analysis, scanning electron microscope (SEM), X-ray diffraction (XRD), and energy-dispersive X-ray (EDAX) analysis.,,,
Antibacterial activity of zinc oxide nanoparticles was tested by disc diffusion assay
The Petri dishes with an average diameter of 60 mm were used in the assay. Petri plates were filled with a medium of Muller-Hinton agar, and then, it is inoculated with the test microorganisms. Sterile discs of 6 mm width were impregnated with 10 μl of various samples (A, B, C, and D, respectively). Prepared plates were kept for 30 min at room temperature, and then, the positive control disc was prepared with 10 μl of amoxicillin which is the standard antibiotic. Finally, all the plates were incubated for 24–36 h at room temperature, and then, the zone of inhibition (mm) was recorded using the millimeter scale.,,,,,,
| Results and Discussion|| |
Qualitative analysis of results of Basella alba extract
The phytochemical components present in the leaf extract of B. alba were investigated by a standard phytochemical screening procedure. Results indicated the occurrence of terpenoids, carbohydrates, glycosides, phenolic compounds, proteins, flavonoids, and phlobatannins in the ethanolic extract (as reducing agents), which plays a synergistic effect in the reduction of zinc in zinc acetate.
Characterization results of Basella alba leave extract-mediated zinc oxide nanoparticles
UV-visible spectra of B. alba extract [Figure 1] show the maximum absorbance at 219 nm, 271 nm, 329 nm, and 338 nm, which is due to unsaturated groups or heteroatoms (C = C, S, N, O) in the extract, and are mainly flavonoids, phenolics, alkaloids, etc. The absorbance at 663 nm is because of chlorophyll (organic chromophores). In the ZnONPs UV spectrum [Figure 2], the maximum absorption peak was observed at 234 nm and 330 nm, which confirms the formation of ZnONPs (due to the surface plasmon resonance effect). FT-IR spectra of plant extracts [Figure 3] reveal the functionalities of OH, C = O, C = C, and C-O, which indicates the presence of phytomolecules. Metal–oxygen (M-O) bond showed FT-IR frequencies below 600 cm−1. The FT-IR spectrum of ZnONPs [Figure 4] absorbed multiple peaks between 433 cm−1 and 548 cm−1. The UV-visible and FT-IR spectrum of B. Alba extract and ZnONPs is shown in [Figure 1].
|Figure 3: Fourier transform infrared spectroscopy spectrum of Basella alba extract|
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|Figure 4: Fourier transform infrared spectroscopy spectrum of Basella alba extract-mediated ZnONPs|
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XRD was used to determine the crystal structure of a material system. The 2Θ values of synthesized zinc nanoparticles [Figure 5] were found to be 31.84°, 34.52°, 36.38°, 47.64°, 56.70°, 63.06°, and 68.10°. The peaks of the graph were matched with the literature (JCPDS File No: 36-1451). Using the Debye–Scherrer equation (Dp = [0.94 × λ]/[β × CosΘ]), the average crystalline size of the nanoparticles was determined. The average crystalline diameters of the formed ZnONPs were found to be 28.65 nm. The obtained XRD plane and data of the B. alba-mediated ZnONPs are similar to the other reported plant extract-mediated (biological method) ZnONPs.
|Figure 5: X-ray diffraction pattern of Basella Alba-mediated ZnO nanoparticles|
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SEM analysis was used to identify the morphology, size, and shape of the nanoparticles. The obtained SEM image showed the spherical shape of obtained ZnONPs [Figure 6]. As the synthesized nanoparticles are spherical, they can easily penetrate the cell wall of the pathogens which contributed to the effective antibacterial activity of the ZnONPs.
|Figure 6: Scanning electron microscope image of Basella alba-mediated ZnO nanoparticles|
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The EDAX investigation was done to check the elemental composition of synthesized ZnONPs. The EDAX results revealed the highest percentages of zinc and oxygen with the elemental composition of 70.04% and 29.96%, respectively, in the B. alba-mediated ZnO [Figure 7].
|Figure 7: Energy-dispersive X-ray analysis elemental analysis of Basella alba-mediated ZnO nanoparticles|
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Antibacterial activity results of Basella alba leave extract, zinc acetate, Basella alba extract-mediated zinc oxide nanoparticles, and standard amoxicillin
B. Alba-mediated ZnONPs (BA-ZnONPs) showed [Figure 8] and [Table 1] well-inhibitory potential against the chosen five bacteria (Gram-positive and Gram-negative bacteria) in comparison with zinc acetate solution and plant extracts. At 10-μl concentration, plant extract does not reveal any inhibition effects on Proteus vulgaris, but BA-ZnONPs exhibit a noticeable effect. Among all the five bacteria, BA-ZnONPs showed the maximum zone of inhibition against Escherichia coli which is a Gram-negative bacterium.
|Figure 8: Antibacterial activity of Basella alba extract, zinc acetate, ZnONPs, and standard amoxicillin on bacterial infection pathogens. Escherichia coli, Enterococcus aerogenes, Pseudomonas aeruginosa, Staphylococcus aureus, and Proteus vulgaris|
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|Table 1: Antibacterial activity of Basella alba extract, zinc acetate, zinc oxide nanoparticles, and standard amoxicillin on bacterial infection pathogens|
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| Discussion|| |
The phytocompounds present in the ethanolic extract of B. alba leaf extract enhance the synthesis of ZnONPs. The presence of functionalities in the plant compounds forms the bond with the metal ions and reduces their size and makes it into nano-sized particles. The alcohols and phenols that are widely seen in secondary metabolite flavonoids were confirmed by qualitative tests. From the literature, it is understood that 70% ethanol extract possesses flavonoid compounds (major phytomolecule) which is also confirmed by qualitative and UV-visible. Further, the XRD analysis proved the spherical crystalline nature of the ZnONPs with an average size of 28.65 nm. The existence of a major elemental percentage of zinc and oxygen without any additional peaks confirmed the purity of the ZnONPs. On anti-bacterial activity, the synthesized ZnONPs revealed the maximum zone of inhibition against the Gram-negative bacteria (E. coli). Xie et al. studied the antibacterial effect of Campylobacter jejuni extract-rooted zinc oxide nanoparticle against Salmonella enterica serovar Enteritidis and E. coli. They found that synthesized particles revealed a noticeable effect and their finding suggest that the microbial effect of ZnONPs resulted from the disruption of the cell membrane and oxidative stress of Campylobacter. Siddiqi et al. summarized the antibacterial effects of ZnONPs; their reviews explain the mechanistic action of ZnONPs toward the bacteria. The authors recorded that the ZnONPs most effectively exhibited inhibitory effects on the microbes such as E. coli, Sarcina lutea, Bacillus subtilis, Pseudomonas fluorescens, Bacillus megaterium, Candida albicans, Staphylococcus aureus, Aspergillus niger, Klebsiella pneumoniae, and Pseudomonas aeruginosa. This microbial effect is driven by the activation of nanomaterials by light and which can penetrate the bacterial cell wall via a diffusion process.
| Conclusion|| |
Green synthesis of nanoparticles used in the research work was reported as nontoxic, ecofriendly, and less usage of earth polluting chemicals than the other physical and chemical methods. Furthermore, the antibacterial activity of BA-ZnONPs has proved that they can act as an antibacterial agent against the chosen infection bacteria. Thus, BA-ZnONPs might be a good alternative to develop as antibacterial agents against the multidrug-resistant strains of bacteria. The applications of BA-ZnONPs may lead to valuable findings in various fields such as medical devices and antimicrobial systems.
Limitation of the study
The above research findings support the green synthesis of zinc oxide nanoparticles which are found to be facile and feasible. However, this synthetic root has shortcomings and challenging issues such as concentration of the B. alba extract, stoichiometric ratios of the extract and precursor, yield and stability of the copper nanoparticles, and other optimization factors such as pH, time, and temperature. All the limitations and shortcomings are can be rectified by the advanced research in nanomaterial preparations.
The authors would like to thank Holy Cross College (Autonomous), Trichy, for providing laboratory facilities to do this research. We also thank CECRI Karaikudi for their technical assistance.
This anti-bacterial activity carried out and their results were approved by the SMS Hospital, Tirucirappalli-620018, Tamilnadu, India.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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