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| ORIGINAL ARTICLE |
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| Year : 2022 | Volume
: 16
| Issue : 4 | Page : 269-273 |
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Exploration of antimicrobial potential of snuhi ksheer according to ksheer sangrahan kaal - A comparative study
Neelam1, Mita Kotecha2, Kamal Nayan Dwivedi3
1 Department of Dravyaguna, NEIAH, Shillong, Meghalaya, India
2 Department of Dravyaguna, NIA, Jaipur, Rajasthan, India
3 Department of Dravyaguna, IMS-BHU, Varanasi, Uttar Pradesh, India
| Date of Submission | 12-Jul-2021 |
| Date of Decision | 12-Oct-2021 |
| Date of Acceptance | 26-Dec-2021 |
| Date of Web Publication | 17-Dec-2022 |
Correspondence Address:
Neelam
Department of Dravyaguna, NEIAH, Shillong, Meghalaya
India
 Source of Support: None, Conflict of Interest: None
DOI: 10.4103/joa.joa_225_21
Introduction: Snuhi, botanically identified as Euphorbia neriifolia L., is a deciduous tree or shrub, belongs to the family Euphorbiaceae, commonly known as milk hedge. Snuhi is one of the constituents of Kshaarasootra, which is used to treat anal fistula. In Charaka Samhita, Snuhiksheera (latex) collection is mentioned specifically at the end of Shishir Ritu (mid-Jan-mid March); on the other hand, the Ksheera collection of any medicinal plant for therapeutic use should be collected in Sharad Ritu (mid-Sept-mid Nov). The study aimed to compare the antimicrobial activity of Snuhi Ksheera collected in Shishirante and Sharadritu. Methods: The antimicrobial efficacy was evaluated against three bacterial strains (Staphylococcus aureus, Streptococcus pyogenes, and Pseudomonas aeroginosa). The study was evaluated using Agar well diffusion method. For samples preparation, 5% solution (ciprofloxacin 500 mg) was used as positive control and 20% dimethyl sulfoxide was used as the negative control. Mueller-Hinton agar medium was used as a culture media for microbes. Results: The results showed that both the ksheera (Shishirante and Sharadritu) have a significant zone of inhibition (ZOI) and Activity Index (AI), but Shishirante collected Snuhi Ksheer have relatively higher ZOI and AI. Hence, both Ksheera can kill the pathogen, but Shishirante collected Snuhi Ksheer showed more potent antimicrobial activity. Conclusion: Antimicrobial activity was better shown by Shishirante collected Snuhi Ksheer against all three microorganisms.
Keywords: Antimicrobial, Sharadritu, Shishirritu, Snuhiksheer
How to cite this article:
Neelam, Kotecha M, Dwivedi KN. Exploration of antimicrobial potential of snuhi ksheer according to ksheer sangrahan kaal - A comparative study. J Ayurveda 2022;16:269-73 |
How to cite this URL:
Neelam, Kotecha M, Dwivedi KN. Exploration of antimicrobial potential of snuhi ksheer according to ksheer sangrahan kaal - A comparative study. J Ayurveda [serial online] 2022 [cited 2023 Feb 6];16:269-73. Available from: http://www.journayu.in/text.asp?2022/16/4/269/364045 |
| Introduction | |  |
Since the time immemorial for the health and well-being of humans, medicinal plants have been the main source of welfare. Valuable drugs are being discovered from time to time based on herbal origin.[1] As per data of the WHO, about 80% of the world population depends on medicinal plant-based drugs for their primary health needs.[2] Active compounds present in plants show different therapeutic qualities/pharmacological actions such as antimicrobial and anti-inflammatory which can be used efficiently to fight the diseases caused by microbes.[3] Due to indiscriminate uses of synthetic antimicrobial drugs, resistance developed in human beings.[4] In addition to resistance, antibiotics are related to severe detrimental results on a host, which include hypersensitivity, allergic response, immune suppression, and depletion of the intestine's ordinary flora. There is a need to mitigate and triumph over antimicrobial resistance and supra infections.[5]
Snuhi (Euphorbia neriifolia L.) is popularly known as “Sehund” or “Thohar” in Hindi, Sudha, and Samantadugdha in Sanskrit. It is considered a small tree or a large shrub up to 6–20 ft. in height. It is found all over India.[6] The plant is pungent, laxative, carminative, improves appetite, beneficial in stomach troubles, bronchitis, tumors, leukoderma, piles, inflammation, anemia, ulcers, fever, and respiration problems.[7] It is Teekshna (stimulant) Virechaka (purgative), Vranaropaka (wound healing), Arshoghna (anti haemorrhoidal), and Kushthaghna (anti dermatosis) Vishahara (anti poisonous).[8] Kshara sutra (alkaline thread) is one of the important preparations of Snuhi ksheera. Snuhi Ghrita, Jatyadi Varti, and Jalodarari rasa are some important formulations of Snuhi.[9]
Snuhi ksheera has substantial capacity and has broad-spectrum activities on numerous ailments. Moreover, their usages had been endorsed in Ayurvedic classic “Charaka Samhita” and different conventional medicinal systems. In Charaka Samhita, the Snuhi ksheera collection is mentioned specifically at the end of Shishir Ritu,[10] on the other hand, the ksheera collection of any medicinal plant for therapeutic use should be collected in Sharad Ritu.[11] However, researches on both the ksheera, which was collected in different seasons, have not been done so far. Hence, both ksheera were selected for the comparative study. Hence, to evaluate both Ksheera experimentally, an in vitro antimicrobial study was carried out against some pathogenic bacteria.
| Materials and Methods | | |
Snuhi ksheera collection and authentication
Snuhi ksheera (Latex of E. neriifolia L-.) were collected from the village of Kunda, Jaipur, Rajasthan. The botanical authentication of plant material was carried out in the herbarium section, Department of Botany, University of Rajasthan, Jaipur with authentication No. RUBL211649 as E. neriifolia L. belongs to the family Euphorbiaceae. For this antimicrobial study, the approval was done by the Institutional Ethical Committee (IEC) with approval No. IEC/ACA/2016/07 on July 15, 2016
Method of collection of Snuhi Ksheera
Latex of Snuhi was drained into an amber color glass bottle by cutting the stem apices with a sharp knife. The cut or slicing immediately oozed out the whitish latex, but oozing did not persist for more than 20 min. Because of air interaction, latex gets solidified in the later stage, so we had added 2 ml of Tween 80 (Polysorbate 80 or Tween 80 is a synthetic nonionic surfactant commonly used in food, cosmetics, and drug formulations as a solubilizer, stabilizer, or emulsifier) after that the latex had collected in the amber color glass bottle. The collected latex was stored and brought to the laboratory for analysis.
Seasons of Snuhi Ksheera collection
Snuhiksheera was collected in two different seasons.
- In Shishirante (end of Shishir Ritu) – 25 February
- In Sharadritu - 15 September.
Selection and collection of pathogens
The pathogenic strains of different species were procured from “Institute of Microbial Technology (IMTech),” Chandigarh, and stock cultures maintenance and antimicrobial study were done at SR Labs, Pratap Nagar, Jaipur. For research work, three bacterial strains such as Staphylococcus aureus (MTCC No. 737), Streptococcus pyogenes (MTCC No. 443), and Pseudomonas aeruginosa (MTCC No. 7925), were taken.
Sample design and sample preparation
- Negative control - 20% dimethyl sulfoxide (DMSO) v/v
- Positive control - 5% solution (ciprofloxacin 500 mg)
- Test sample - Snuhiksheera of two different seasons
- Test sample 1 - Snuhiksheera of Shishirante
- Test sample 2 - Snuhiksheera of Sharadritu.
Extract solution preparation and test procedure
An in vitro experiment was performed by dissolving each extract (0.2 g) in 0.1 ml of DMSO and volume was made up to 1 ml with distilled water to prepare 200 mg/ml w/v solution. The 20% DMSO alone did not show any biological activity, hence used as a negative control. As standard antibiotics, ciprofloxacin (5 μg/ml) was used and prepared a 5% solution. For media preparation, Muller-Hinton agar (3.8 g/100 ml) was dissolved in distilled water in the conical flask and sterilized in an autoclave for 30 min; pour the molten agar into plates. After solidification, wells were made by using 5 mm of steel borer. For the determination of antimicrobial activity, Agar well diffusion method was used.[12] The four wells (diameter 5 mm) were used and the volume applied in each well is 100 μl. One well for a standard antibiotic was used as a positive control, another two well-containing samples (aqueous and ethanolic solution), and the fourth well containing 20% DMSO for negative control.[13] The experiment was carried out in triplicate and the average diameter of zone of inhibition (ZOI) was measured in mm, with the help of a scale. Then, the mean was calculated of the three readings taken and the activity index (AI) was calculated. AI was done to find out where test samples stand in comparison to the standard antibiotic.
Determination of activity index
The AI of the test sample was calculated as[14]
AI = ZOI of the extract
ZOI obtained by standard drug
AI - Test substance >0.50 is considered a significant activity.[15]
| Results and Discussion | | |
In the present study, the bioassay result of antimicrobial activity of both (test samples 1 and 2) are tabulated in [Table 1] and [Table 2]. It was evident from the result that both the samples were found active against the bacterial strains. Concerning standard antibiotic (positive control), the ethanolic extract of test sample 1 was found a privileged ZOI against all bacterial strains when compared to the aqueous extract of test sample 1 [Figure 1]. Same in the case of test sample 2, i.e., the ethanolic extract showed a better ZOI than aqueous extract [Figure 2] and [Figure 3]. However, the degree to inhibit the growth of bacteria was significantly found in test sample 1 [Figure 4]. The AI of the test substance >0.5 is considered a significant activity. The ethanolic extract of test sample 1 [Figure 5] and [Table 3] showed a significant AI against all three bacterial strains (S. aureus 0.85, S. pyogenes 0.52, and P. aeruginosa 0.77), but the aqueous extract showed activity against only two bacterial strains (S. aureus 0.65 and P. aeruginosa 0.61). The aqueous and ethanolic extracts of test sample 2 [Table 4] showed better AI against S. aureus and P. aeruginosa) which again proved the significant antimicrobial efficacy of test sample 1 than test sample 2. | Table 1: Zone of inhibition of test sample 1 with positive and negative control
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 | Table 2: Zone of inhibition of test sample 2 with positive and negative control
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 | Figure 1: The antimicrobial activity of test sample 1 with positive and negative control. Zone of inhibition of ethanolic extract (14 mm) of Shishirante collected Snuhi ksheera showed maximum antimicrobial activity against Pseudomonas aeruginosa and to some lesser extent against Staphylococcus aureus and Streptococcus pyogenes in comparison to standard drug
Click here to view |
 | Figure 2: The antimicrobial activity of test sample 2 with positive and negative control. ZOI of ethanolic extract (13 mm) of Sharad ritucollected Snuhi ksheera showed maximum antimicrobial activity against Pseudomonas aeruginosa and to some lesser extent against Staphylococcus aureus and Streptococcus pyogenes in comparison to standard drugs
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 | Figure 3: Antimicrobial activity of test sample 2 against target microorganisms The activity of Staphylococcus aureus, Pseudomonas aeruginosa and Streptococcus pyogenes
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 | Figure 4: Comparative activity index of test sample 1 and test sample 2. (a) Ethanolic extract (0.85) of Shishirante collected Snuhi Ksheera (test sample 1) showed maximum activity index against Staphylococcus aureus, 0.77 and 0.52 against Pseudomonas aeruginosa and Streptococcus Pyogenes, respectively. Activity index of Aqueous extract of test sample 1 showed maximum activity (0.65) against S. aureus, 0.61 and 0.42 against P. aeruginosa and S. pyogenes, respectively. (b) Ethanolic extract of Sharad Ritu collected Snuhi Ksheera (test Sample 2) showed maximum Activity Index (0.78) against S. aureus, 0.72 and 0.47 against P. aeruginosa and S. pyogenes, respectively. Activity index of aqueous extract of test sample 2 showed maximum activity (0.65) against S. aureus, 0.61 and 0.42 against P. aeruginosa and S. pyogenes, respectively
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 | Figure 5: Antimicrobial activity of test sample 1 against target microorganisms The activity of Staphylococcus aureus, Pseudomonas aeruginosa and Streptococcus pyogenes
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Probable mode of action according to Ayurveda perspective
There are six Ritus (seasons) and accordingly six Rasas (taste) also. Acharya Vagbhata highlighted the relation between seasons, Panchamahabhuta (five basic elements), the composition of Rasas (taste), and evaluation of Rasas (taste).[16] He further said that certain Rasas (taste) will form in other seasons also, but the formation of Rasas (taste) in the specified seasons will be the best.
Vayu and Aaksha both having Ruksh (Dryness), Laghu (lightness), and Sukshma (fineness) Guna, which helps in the depletion of moisture, pus sweat, etc.; hence, it inhibits bacterial growth in wounds also through Sukshma Guna the drug can penetrate through the minutest channels of the body.
Snuhi itself has Katu (bitter) Rasa having the property of Krimihar (antibacterial, Kandughna (antipruritic), Vrana Ropana (healing), Mamsa Lekhan (scraping), etc.[18] Hence, the Ksheera, which was collected at the end of Shishir Ritu will potentiate the Krimihar (antibacterial) properties. Hence, Shishir Ritu collected Snuhi Ksheera showed significant antimicrobial activity than Sharad Ritu.
| Conclusion | | |
From the above results and discussion, it can be concluded that the Snuhi Ksheera (Latex of Euphorbia nerifolia L.), which was collected at the end of Shishir Ritu possess significant antimicrobial activity against all three bacterial strains (S. aureus, S. pyogenes, and P. aeruginosa) then Sharad Ritu collected Snuhi Ksheera. This antimicrobial property is surely due to some antimicrobial substances in Ksheera (latex). The limitation of the study was the collection of Snuhi Ksheera. The collection was much difficult unless added Tween 80. Hence, the future study may be carried out to explore new techniques to collect the Ksheera (latex) from any medicinal plant without adding an emulsifier (Tween 80) and to search the lead component responsible for this antimicrobial activity.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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