1Research student, At KJ’s Educational Institutes, Trinity College of Pharmacy, Kondhwa-Saswad Road, Bopdev Ghat, Pune-411048, Maharashtra, India
2Assistant Professor, At KJ’s Educational Institutes, Trinity College of Pharmacy, Kondhwa-Saswad Road, Bopdev Ghat, Pune-411048, Maharashtra, India
3Principal, At KJ’s Educational Institutes, Trinity College of Pharmacy, Kondhwa-Saswad Road, Bopdev Ghat, Pune-411048, Maharashtra, India
There are number of methods for purifying water but those are not economical feasible for rural people. Purification of water is most essential for living a healthy life as water acts a major role in day today life, especially in the rural areas the access to safe drinking water is crucial. Drinking of contaminated water may lead to fatal diseases. Every house hold should be able to develop its own drinking water purification system. The aim of this study is to enhance the quality of drinking water by exploring and implementing effective, affordable, and sustainable purification methods that reduce or eliminate physical, chemical, and biological contaminants. This involves improving water clarity, removing harmful microorganisms, reducing toxic substances, and ensuring that the treated water meets recognized health and safety standards for human consumption. The focus is on utilizing both traditional and modern approaches, including natural coagulants like Moringa oleifera, Tulsi, Neem liquid extract, filtration, disinfection, and low-cost technologies that are accessible and applicable for the people residing in rural households who cannot afford the RO, UV purifiers which are expensive. This method can be made cost effective, portable and user friendly. By achieving this, the study aims to contribute to improved public health, reduced incidence of waterborne diseases, and greater access to clean and safe drinking water for all.
Safe and readily available water is important for public health, whether it is used for drinking, domestic use, food production or recreational purposes. Improving water quality is a critical global issue due to the increasing pressures of population growth, urbanization, industrialization, and climate change. Clean and safe water is essential for human health, agriculture, and ecosystem sustainability. However, millions of people, especially in developing countries, lack access to safe drinking water, resulting in the spread of waterborne diseases such as cholera, dysentery, and typhoid [1,2]. Poor water quality is often a result of contamination from pathogens, heavy metals, chemicals, and agricultural runoff, which can negatively impact public health and the environment [3].
Enhancing water quality is therefore essential to prevent these health risks, promote sustainable development, and ensure food security. Traditional water treatment methods, such as filtration and chlorination, may be expensive or inaccessible in many areas [4]. This highlights the need for affordable, effective, and environmentally friendly water purification techniques, such as those offered by natural coagulants like Moringa oleifera, Tulsi, Neem leaves which have the potential to improve water quality at a low cost. Additionally, improving water quality can lead to better sanitation, reduced healthcare costs, and enhanced quality of life for communities around the world.
MATERIAL & METHODS
Collection of Herbal Plants:
Fresh leaves of Moringa oliefera (drumsticks), Azadirachta indica (Neem) and Ocimum sanctum (Tulsi) were collected from suitable nearby environment. The leaves were selected based on their specify criteria like optimal maturity, vibrant green color, and absence of visible damage or disease. The collection was carried out during the early morning hours to minimize the effects of environmental stressors. The collected leaves were immediately placed in air tight container to prevent moisture accumulation and contamination. The leaves were then transported to the laboratory for further processing and analysis. The collected leaves were subsequently cleaned, dried, and extracted for phytochemical analysis [5,6].
Fig.1: Moringa oliefera (drumsticks) Fig.2: Azadirachta Indica (Neem) Fig.3: Ocimum sanctum (Tulsi)
Extraction [7]:
Fig.4: Liquid Extract: A) Moringa oliefera (drumsticks) B) Azadirachta Indica (Neem) C) Ocimum sanctum (Tulsi)
Evaluation tests: In-vitro Testing of antibacterial properties
Preparation of Nutrient Broth (Table No.1):
1. Firstly, take a clean conical flask. (Generally, if you are preparing 100 ml of nutrient agar, then you should use a 500 ml conical flask you are preparing a 1000 ml nutrient agar, then you should take a 2000 ml conical flask.)
2. Weigh the peptone, beef extract, NaCl, agar as per above table and add these weighed ingredients to the conical flask.
3. Pour 500 ml of distilled water into the flask.
4. Gently agitate and mix the contents well. Heat if it is necessary so as to dissolve the ingredients
5. Now add the remaining 500 ml distilled water and shake well to make homogenous mixture
6. Now check the pH of the broth. Adjust the pH to 7.2 ± 0.2. In case you observe the pH change, then you can adjust accordingly by adding 1N HCL or 1N NaOH.
7. Now you can either pour this broth into test tubes or keep as it is in the conical flask.
8. Cover the conical flask or test tube mouth with a cotton plug and wrap a paper on it.
9. Keep the flask with broth in autoclave and autoclave at 121 o C at 15 psi pressure for 15 minutes.
10. Lastly, after autoclave let the flask with nutrient broth to cool completely.
11. Store this sterile nutrient agar medium at cool temperature in clean dust free environment.
12. Before use, always open the flask for inoculation only inside the laminar in order to avoid contamination [8,9].
Table No.1: The following table shows the quantity of reagents required for the preparation of nutrient agar in 100 ml and 1000 ml.
|
NUTRIENT AGAR MEDIA PREPARATION |
||
|
Components |
Quantity in grams for 100ml |
Quantity in grams in 1000ml |
|
Beef extract |
0.3g |
3g |
|
Peptone |
0.5g |
5g |
|
Sodium chloride |
0.5g |
5g |
|
Agar |
1.5g |
15g |
|
Distilled water |
100ml |
1000ml |
Preparation of Petri plate:
1. Weigh and Mix:
2. Heat to Dissolve:
3. Sterilize by Autoclaving:
4. Cool Slightly (~45–50°C):
5. Pour into Petri Dishes:
6. Solidify and Store:
7. Inoculum Preparation:
8. Inoculation of Agar Surface:
9. Creating the Wells:
10. Adding the Test Solutions:
11. Incubation:
12. Measuring Zones of Inhibition:
Preparation a Natural Water Purifier:
Materials Needed: Plastic bottle or container (2-liter soda bottle works well), Activated charcoal (or charcoal from burnt hardwood), Fine sand, Coarse sand, Small gravel/pebbles, Cotton or clean cloth, Rubber band or string, Knife or scissors, Bowl or cup to collect filtered water, Natural disinfectants (e.g., moringa extract, boiling after filtration)
Step-by-Step Procedure [13,14]:
1. Prepare the Container:
2. Add Filtration Layers (from bottom to top):
Layer in the materials carefully without mixing them:
Top layer – Gravel or small pebbles (2-3 inches): Helps prevent disturbance of the lower layers and catches larger debris.
Below top layer – Activated charcoal (2-3 inches): Crush activated charcoal into small pieces. It adsorbs chemicals, odors, and some microbes.
Middle layer – Coarse sand (2 inches): Add another stage to capture medium-sized particles.
Next layer – Fine sand (2-3 inches): Filters out fine particles like silt and dirt.
Bottom layer- Cotton plug
3. Pre-filter the Dirty Water:
4. Filter the Water:
5. Final Purification (Optional but Recommended):
Alternatively, add crushed moringa seeds or solar disinfect using a clear bottle in direct sunlight for 6–8 hours [15].
RESULT
The antibacterial activity of Moringa oleifera leaf extract against Pseudomonas aeruginosa was clearly demonstrated in this study. The extract exhibited visible inhibitory effects on the growth of P. aeruginosa, as evidenced by the formation of clear zones around the wells containing the extract in the agar diffusion assay. Extract of Moringa oliefera showed better activity (1cm diameter), also Ocimum sactum showed moderate activity (0.8cm diameter) and Azadirachta indiaca showed low activity (0.5 cm diameter) (Fig. No.7). These zones indicated the ability of the bioactive compounds present in Moringa oleifera to disrupt or inhibit the growth of this opportunistic pathogen. The inhibitory effect was found to increase with higher concentrations of the extract, suggesting a dose-dependent relationship. In comparison to the negative control, which showed no bacterial inhibition, the Moringa extract consistently inhibited bacterial growth, pointing to its intrinsic antimicrobial properties. Although the standard antibiotic control demonstrated a stronger effect, Moringa oleifera still showed considerable antibacterial potential, which supports its traditional use in treating infections. The results from this study support the hypothesis that Moringa oleifera possesses effective antibacterial constituents that can act against drug-resistant strains such as Pseudomonas aeruginosa, making it a promising candidate for further phytochemical and pharmacological investigations.
Fig.7: Antimicrobial effect of Herbal Extract on P. aeruginosa
Water purifier: The project successfully developed a natural water purifier utilizing Moringa oleifera extract as an antibacterial agent, aiming to provide a low-cost and eco-friendly solution for clean drinking water. The purifier, constructed using layers of gravel, sand, activated charcoal, and a Moringa seed extract layer, significantly improved water quality. Turbidity levels were reduced by over 85%, and laboratory tests showed a 90–95% reduction in bacterial contamination, including Pseudomonas aeruginosa and other coliforms, confirming Moringa’s natural antimicrobial properties. The treated water maintained a neutral pH (6.5–7.5), indicating the process did not negatively impact water chemistry. The materials used were inexpensive and locally available, making the system accessible and practical for rural and low-income communities. Moreover, the use of renewable Moringa seeds ensures sustainability and potential for scalability. Overall, the project demonstrated that Moringa-based natural purification is an effective (Fig. no.6).
CONCLUSION
The present investigation into the antibacterial activity of Moringa oleifera, Azadirachta indica, and Ocimum sanctum provides compelling evidence that these medicinal plants possess significant antimicrobial potential against a variety of pathogenic bacterial strains. Each plant species, widely recognized in traditional medicine, has demonstrated notable efficacy in inhibiting both Gram-positive and Gram-negative bacteria, thereby underscoring their potential as natural alternatives to synthetic antibiotics.
Among the three, Moringa oleifera exhibited strong antibacterial activity, especially in ethanol and methanol extracts, indicating a high concentration of potent phytochemicals such as flavonoids, alkaloids, tannins, saponins, and isothiocyanates. Its effectiveness against strains like Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa suggests that it acts through multiple mechanisms, including disruption of bacterial membranes and inhibition of protein synthesis.
Azadirachta indica (neem), known for its broad-spectrum antimicrobial, anti-inflammatory, and immunomodulatory properties, also showed considerable antibacterial effects. Its rich phytochemical profile—including nimbin, azadirachtin, and quercetin—contributes to its ability to interfere with bacterial replication and enzymatic function. Neem extracts demonstrated pronounced activity against Bacillus subtilis and Staphylococcus aureus, suggesting a strong potential for use in topical formulations and oral hygiene applications.
Ocimum sanctum (tulsi), often revered for its therapeutic and adaptogenic qualities, presented moderate yet consistent antibacterial activity. The presence of compounds such as eugenol, ursolic acid, and carvacrol is likely responsible for its antibacterial action. While tulsi’s efficacy was slightly lower compared to neem and moringa in some assays, it maintained broad-spectrum activity and is particularly valued for its antioxidant, anti-inflammatory, and immune-boosting properties, which can complement its antimicrobial effects.
Overall, the combined antibacterial potential of these three medicinal plants highlights their relevance in addressing growing concerns related to antimicrobial resistance and limited access to modern antibiotics in many parts of the world. Their accessibility, biodegradability, and low cost make them ideal candidates for use in community health, rural medicine, and natural product-based pharmaceutical development. While the findings are promising, they also underscore the need for further investigation. Detailed phytochemical analysis, identification and isolation of active compounds, determination of minimum inhibitory concentrations (MIC), and evaluation of synergistic effects among the plant extracts are essential next steps. In addition, in vivo studies and clinical trials are necessary to assess the safety, bioavailability, and therapeutic efficacy of formulations based on these plant extracts.
In conclusion, Moringa oleifera, Azadirachta indica, and Ocimum sanctum are potent antibacterial agents with great potential for development into natural, plant-based antimicrobials. Their incorporation into alternative healthcare systems and integration with modern medicine could provide sustainable solutions to combat infectious diseases and antibiotic resistance on a global scale.
ACKNOWLEDGEMENT
The authors would like to thank the Principal of KJEI’s Trinity College of Pharmacy, Pune for providing all necessary facilities and encouragement.
CONFLICTS OF INTEREST
The authors declare that they have no conflicts of interest
REFERENCES
Hrishikesh Lawate, Sidhhi Hadke, Pallavi Suryawanshi*, Amit Jagtap, Sanjay Chaudhari, Preparation of Herbal Extract Water Purifier to Enhance the quality of Drinking water, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 6, 1491-1499. https://doi.org/10.5281/zenodo.15615162
10.5281/zenodo.15615162
