AADHAVAN.T.

M, Guided By:Karthick Raja

Namasivayam
EFFECT OF PHYTO INSPIRED ZINC OXIDE NANOPARTICLES ON THE ANTIMIROBIAL
EFFICIENCY AGAINST MICROBIAL STRINESS ASSOCIATED WITH SKIN INFECTION

INTRODUCTION
Skin infections caused by pathogenic bacteria are a significant global health concern, and the increasing
incidence of antibiotic-resistant strains has further complicated the treatment options. Traditional antibiotics have
become less effective against resistant strains, making it challenging to manage skin infections effectively. In
recent years, there has been growing interest in developing alternative antimicrobial agents that can overcome
the limitations of traditional antibiotics. Nanoparticles have emerged as a potential alternative to antibiotics due to
their unique physicochemical properties and broad-spectrum antimicrobial activity. Zinc oxide nanoparticles (ZnO
NPs) have received significant attention due to their biocompatibility, stability, and low toxicity.

In recent years, there has been an increasing focus on developing green and sustainable synthesis methods for
nanoparticles. Phyto-inspired ZnO NPs synthesized from natural plant extracts have shown promising
antimicrobial activity. The phytochemicals present in the plant extracts can serve as reducing agents, stabilizers,
and capping agents during the synthesis process, which can improve the stability and biocompatibility of the
resulting nanoparticles. In addition, phyto-inspired ZnO NPs have been reported to exhibit enhanced antimicrobial
activity compared to their conventionally synthesized counterparts.

MATERIALS AND METHODS

ANTIFUNGAL
➢ The strain was cultivated on lab-rock and kept alive using
Sabouraud Maltose Yeast Extract broth.
➢ A loopful of the fungal culture was inoculated in a 250 ml
Erenlymer flask with 100 ml of sterile SMYB broth to
prepare the inoculum.
➢ Cells were extracted by centrifugation at 3000 rpm for 15
min while the flask was incubated at 300 C for 7 days on a
rotary shaker at 150 rpm.
➢ The resulting biomass was dried for 24 hours in an oven
at 600C.
ANTIBACTERIAL
➢ For this study, laboratory stock cultures of Escherichia coli, Bacillus
subtilis, and Staphylococcus aureus were employed.Nutrient agar
slants were used to sustain all of the strains.
➢ To prepare the inoculum, 50ml of sterile nutritional broth was
inoculated with a loopful of the appropriate strains under aseptic
conditions, and the mixture was then incubated at 37°C under
shaking conditions for 16 hours.
➢ On sterile nutritional agar media, 100 l of the prepared bacterial
inoculum were applied.
➢ Aliquots of the plant composite were dissolved in 10% DMSO and
applied to agar cups (8 mm) using sterile well punctures.
➢ The agar cups were then incubated at 37 °C for 24 hours. The
zone of inhibition was noticed following the incubation period.
➢ The common antibiotic served as a good control.

RESULTS
ANTIFUNGAL
It was investigated for antifungal activity against fungal strains
Candida albicans. Among control and silver-treated catheters,
maximum inhibition was recorded at silver-treated catheters
against all the fungal strains. The zone of inhibition against
Candida albicans was 19 mm.
ANTIBACTERIAL
Evaluation of the antibacterial activity of zinc oxide was
recorded. Among different concentrations, maximum
inhibition was recorded at 500 µg/ml against all the bacterial
strains. In zone of inhibition against Escherichia coli was 19
mm in 500 µg/ml.

DISCUSSION AND CONCLUSION

The results of this study suggest that phyto-inspired ZnO nanoparticles exhibit potent antimicrobial activity against microbial
strains associated with skin infections. The synthesized nanoparticles showed high efficacy in inhibiting the growth of both Gram-
positive and Gram-negative bacteria, including Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, which
are common pathogens responsible for skin infections.
The enhanced antimicrobial activity of phyto-inspired ZnO NPs can be attributed to the presence of phytochemicals in the plant
extracts used for synthesis. These phytochemicals can serve as reducing agents, stabilizers, and capping agents during the
synthesis process, which can improve the stability and biocompatibility of the resulting nanoparticles. Furthermore, the
phytochemicals present in the plant extracts can interact with the bacterial cell membrane and disrupt its integrity, leading to cell
death.

In conclusion, our findings suggest that phyto-inspired ZnO NPs have the potential to be developed as a sustainable and effective
antimicrobial agent for the treatment of skin infections. The use of green and sustainable synthesis methods for nanoparticles can
help overcome the limitations of traditional antibiotics and reduce the risk of antibiotic resistance. Further studies are warranted to
evaluate the toxicity and biocompatibility of these nanoparticles in vivo and to optimize their properties for clinical use.

BIBLIOGRAPHY
❖ Gupta, A., Qureshi, M., Singh, S., Kumar, S., Kumar, R., Koul, V., & Singh, K. (2021). Phyto-inspired synthesis of zinc
oxide nanoparticles and their antibacterial activity. Journal of Microbiology, Immunology, and Infection.
❖ Saeed, S., Idrees, M., & Arshad, M. (2019). Green synthesis and characterization of ZnO nanoparticles using the
aqueous extract of Fagonia indica and their potential antimicrobial activity. Materials Science and Engineering.
❖ Ghorbani, H. R., Zolfaghari, B., Mirzaei, A., & Ghafourian, S. (2021). Green synthesis of zinc oxide nanoparticles: a
review of antibacterial, antifungal, and antiviral activities. ChemistrySelect.
 Students Name: Aadhavan.T.M,
Guided By: Dr.Karthick Raja Namasivayam

SCREENING OF METAL OXIDE NANOPARTICLES INDUCED

HEMOLYTIC EFFECT - AN IN-VITRO STUDY

INTRODUCTION
In-vitro studies can be useful for assessing the potential toxicity
of metal oxide nanoparticles (MONPs) on red blood cells. One
common way to evaluate the hemolytic effect of nanoparticles is
by performing a screening assay, which measures the release
of hemoglobin from erythrocytes following exposure to MONPs.
To perform the screening assay, a sample of whole blood is
collected from a donor and then centrifuged to obtain a red blood
cell (RBC) suspension.

MATERIALS AND METHODS

Biocompatibility of respective nanoformulation was further confirmed by hemocompatibility
assays using peripheral blood cells. 5 mL of fresh blood sample was taken from the healthy
volunteer. 500µL of the collected blood was transferred to an equal volume of isotonic saline
e
followed by the addition of 50, 100, and 250 µL of samples were added. Respective treatment
groups were incubated at 35 °C for 12 h. After the incubation period, plasma was collected by
centrifuging the sample. Blood smears were also prepared from the respective treatment group
and stained with Leishman staining. After the staining, slides were observed under a microscope
to observe blood cell lysis, morphological changes, or abnormalities.

RESULTS
Haemocytes are readily obtainable and exhibit highly observable morphological changes to the
exposure of the text compound which can be utilized in the toxicity assessment system. In the
present investigation, human erythrocytes were used for the determination of hemolysis.
Hemocompatibility studies are studied by determination of hemolysis at time dependent intervals
Significant effect was not observed at all the tested time periods in all the treatment. Microscopic
examination of blood smears also revealed no morphological changes and lysis of haemocytes.
As in control, all the treatment groups exhibited normal cellular morphology of haemocytes.

DISCUSSION AND CONCLUSION

Biocompatibility of respective treatment was further confirmed by in vitro hemolysis assay using
peripheral blood cells Toxicity assessment under in vitro condition using haemocytes have used
for the decades. All the findings clearly shows the high level of safety and biocompatibility of
nanoparticles.

Control A B C

BIBLIOGRAPHY
• Karthick Raja Namasivayam, S., Arvind Bharani, R., & Karunamoorthy, K. (2018, December).
Insecticidal fungal metabolites fabricated chitosan nanocomposite (IM-CNC) preparation for
the enhanced larvicidal activity - An effective strategy for green pesticide against economic
important insect pests. International Journal of Biological Macromolecules, 120, 921–944.
https://doi.org/10.1016/j.ijbiomac.2018.08.130
• In-Text Citation: (Karthick Raja Namasivayam et al., 2018
• Faria N, Song H, Branquinho R, et al. Hemolytic activity of metal oxide nanoparticles - a
systematic review. Nanotoxicology. 2019;13(4):466-492. doi:10.1080/17435390.2019.1599508
• Hamedi A, Yazdanparast R, Shahverdi AR. In vitro hemolytic activity of ZnO and CuO
nanoparticles on human peripheral blood erythrocytes. J Biomed Mater Res A.
 Students Name: Aadhavan.T.M,
Guided By: Dr.Karthick Raja Namasivayam

NON TARGET TOXICITY ASSESSMENT OF BIODEGRADIZED

PRODUCTS OF MICROPLASTIC USING ZEBRA FISH MODEL

INTRODUCTION
Microplastics are tiny plastic particles that are less than 5 mm in size, which are
found in various environmental media such as oceans, freshwater, soil, and air.
Microplastics are generated by the fragmentation of larger plastic items, such as
packaging materials, and from the shedding of microplastic fibers from textiles during
laundering. Biodegradation is considered as one of the promising ways to mitigate
microplastic pollution. However, there is a lack of knowledge on the potential adverse
effects of biodegraded microplastic products on non-target organisms.

MATERIALS AND METHODS

Collection of biodegraded product of microplastics:
Microplastics samples that degraded with bacterial inoculant (rhizobium) was used in this study.
Zebrafish embryonic toxicity study:
Collection and maintenance of zebrafish embryo under standard condition
e
Addition of microplastic (0.1ml) to the culture medium

Determination of various parameters at predetermined time interval (24,48,72,96,120) using

phase contrast microscope

Parameters studied (mortality of embryo, mobility pattern, hatching rate %, morphological

changes)

RESULTS
The study is elongated to 96 hours since embryogenesis
occurs at 96 hours and the four stages of embryonic
development occur between 24 and 96 hours
(Figure 8: a-h). In both the control and nanoparticle-
treated flasks, hatching, early larval stages, segmented
embryos, and pharyngeal embryos were visually
perceived. A substantial vicissitude in embryonic Figure 8.Effect of biodegraded products of microplastics on developmental neurotoxicity in Zebrafish

morphology was not optically discernible in the test group of zebrafish (Figure 8). No
toxicity was visually perceived in the treatment groups. The control embryos were
mundane. The heart rate of the rats in both tests and the control group was
mundane, ranging from 160 to 180 beats per minute. The motility pattern in the
experiments (Figure 8: e-f) was the same as the movement pattern in the reference,
indicating that the metabolites created by the nanotechnology did not cause any
discernible toxicity.

DISCUSSION AND CONCLUSION

Because of the increased use of novel medications, there is an urgent need for low-
cost, rapid in vivo models to investigate total drug toxicity and metabolic effects. One
such model is the zebrafish, which has many similarities to humans (Zhao et al 2022)
Because nanoparticles may cross the blood-brain barrier, they have prompted fresh
concerns regarding potential neurotoxicity. The zebrafish offers significant tools for
overcoming the limitations of other models in neurotoxicity testing. However, the
utilization of early life stages in humans and animals to estimate its long-term toxicity
is restricted. In this light, the zebrafish has been regarded as an effective model
organism for ketamine biosafety studies

BIBLIOGRAPHY
• Theodorou E, Karaolia P, Anastasiadou K, Hapeshi E, Christou A, Torres T, et al. Toxicity of
biodegraded microplastics and nanoplastics in a zebrafish embryo model. Nanomaterials.
2021;11(1):148.
• Rowe SP, Lindeque PK, Galloway TS. The impact of microplastic biodegradation on toxicity to
the marine fish Danio rerio. Environ Sci Technol. 2020;54(17):10998-11007.
• Kim T, Choi K, Lee J, Kang JH, Kim SY, Kim SW, et al. Biodegradation of microplastics in
marine environments: a review on current knowledge and future perspectives. Marine Pollution
Bulletin. 2019;141:448-458.