ISSN: 2322 - 0902 (P)

ISSN: 2322 - 0910 (O)

International Journal of Ayurveda
and Pharma Research

Research Article

AN EXPERIMENTAL STUDY ON THE EFFICACY OF VAGBHATOKTA JALA NIRVISHIKARANA YOGA

ON POLLUTED WATER
Shivanand B. Kalasannavar1*, S. K. Hiremath2, U. S. Hampannavar3
*1Assistant Professor, Department of Agada Tantra, Govt. Ayurveda Medical College, Mysore, Karnataka, India.
2Professor and Head, Department of Agada Tantra, Taranath Govt. Ayurveda Medical College, Bellary,

Karnataka, India.
3Principal, Maratha Mandal Engineering College, Belagaum, Karnataka, India.

ABSTRACT
Background: Increased suffering and death due to impure water consumption, along with the failure
of the outreach of advanced techniques to the deprived people with middle socioeconomic status, has
directed the evaluation of efficacy of the classical Ayurvedic formulations used in water purification.
Aim: To evaluate the efficacy of Vagbhatokta Jala Nirvishikarana Yoga in terms of its physiochemical
and antimicrobial qualities for the treatment of polluted river water.
Materials and Methods: The Jala Nirvishikarana Yoga (JNY) was prepared according to the classical
method, and analytical study was performed. Optimum dose and period for JNY was calculated, and
the water sample was treated accordingly. The sample was analysed for various physical, chemical
and microbiological parameters before and after treatment. Stability and efficacy of the formulation
was determined after 3 months and 1 year. Results: JNY is a thick, alkaline and viscous liquid bearing
a characteristic taste and pleasant odour. The optimum dose and period was found to be three drops
per 100 mL for 30 min of contact time. No significant change in the physiochemical parameters was
observed except pH. Apart from the variation in pH, JNY was observed to have antimicrobial activity.
Reduction in the coliforms and inhibition of bacterial and fungal colonies in the sample were
observed. JNY was observed to be stable for 1 year at room temperature. Conclusion: JNY showed
efficient antimicrobial activity and was stable and effective for at least 1 year at room temperature.
However, in larger doses, its administration resulted in increase of pH of water.
KEYWORDS: Jala Nirvishikarana Yoga, Stability, Antimicrobial activity, pH, Dose, Period.
INTRODUCTION
As a result of industrialization, pollution of monitor water quality provided for the purpose of
the natural resources has been one of the major drinking, irrigation etc.
concerns of the modern world. Among them, water The quality of water is usually analysed by
pollution poses a huge risk to the well-being of the considering various parameters, such as
humans, as it is the most common source of temperature, dissolved oxygen level, conventional
diseases.[1] Rivers are the most susceptible of variables (pH, total dissolved solids, conductivity),
contamination, as they are known to be the carriers and the concentration of nutrients, metals and
of agricultural run-offs and wastes from the industrial chemicals. However, microbiological
municipalities and industries.[2] assessment of water source is done by the detection
Foul smell and presence of toxic substances of indicators such as faecal coliforms, faecal
are often the major concerns of the inhabitants living enterococci and streptococci, sulphur-reducing
on the banks of polluted rivers.[3] Worldwide, clostridia etc.[5] The faecal contamination of water is
approximately 5 million of people or more have usually determined by detecting the presence of
perished because of water-borne diseases[1]. faecal coliforms by most probable number (MPN) and
Annually, approximately 37.7 million people in India membrane filtration method.[6]
are known to be affected by water-borne diseases. [4]
Polluted water is usually treated chemically
Hence, the foremost objective of the environment and biologically before their release in the water
protection agency should be to minimise the bodies. Purification of water by implementing the
frequency of pollution-oriented problems and traditional and ancient method of India, i.e. by use of

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 Shivanand B. Kalasannavar et al. Efficacy of Vagbhatokta Jala Nirvishikarana Yoga on Polluted Water
herbs and shrubs, helps to cleanse the water while nitrate). All the tests were performed as prescribed
retaining its natural benefits.[7] As such, by the Ayurvedic Pharmacopeia of India.
environmental toxicology has been known to come Optimum dose and period
under the purview of Agada tantra[8] and thus, has a The optimum dose and optimum period of
wide scope of research. JNY application was determined by adding JNY to the
Therefore, the following study was conducted river sample in required quantity and for requisite
to evaluate the efficacy of an Ayurvedic formulation, time, respectively. Bureau of Indian Standards
Vagbhatokta’s Jala Nirvishikarna yoga, in terms of its methodology was followed for conducting the
physiochemical and antimicrobial qualities for the analysis.
treatment of polluted river water. The optimum dose of JNY was fixed by taking
MATERIALS AND METHODS river water sample (100 mL) in a set of four beakers.
Source of sample One among the sets was kept as control, whereas
The study was conducted at the Department 0.25 mL (five drops), 0.5 mL (10 drops) and 1 mL (15
of Agada Tantra, KLE University’s Shri BMK Ayurveda drops) of JNY were added to the rest of the beakers,
Mahavidyalaya, Belgaum. The water sample was respectively. Similar set of beakers were used for
collected from Malaprabha river at Kadrolli , Belgaum determining the optimum period of JNY. Following
(located between 15.6976 ̊ North latitude and the addition of the optimum dose of JNY, the sample
74.7352°East) by following standard operating in the beakers were incubated for 30 min, 1 h and 2 h,
procedures. respectively. The samples were then analysed to
Jala Nirvishikarana yoga determine the optimum period of JNY.
Jala Nirvishikarana Yoga (JNY), a novel Stability study
formulation, was prepared by adding 300 g of each The stability of JNY was determined by
drug: Ajashrungi (Pistacia chinensis), Indravaruni storing the formulation in sterilised air-tight glass
(Citrullus colocynthis), Uttamarini (Daemia extensa), bottles for a period of 1 year under aseptic condition.
Guduchi (Tinospora cordifolia), Phanijjaka (Ocimum One of the bottles were kept at room temperature
basilicum) and Ativisha (Aconitum heterophyllum) in whereas the other was stored under cold condition.
the form of ash and paste of extra ingredients Periodical testing of the stored JNY was done for
(Prativapa dravya) into distilled water in a fixed ratio above mentioned organoleptic parameters and pH.[9]
as shown in Table 1. The ash was adequately cooled Microbiological parameters
till it turned white and analysed by Rekhapurnatvum Microbial limit test
test. Prativapa dravya was prepared by grinding and 1. Most probable number
mixing of Sarala Niryasa (Pinus roxburghii), Haridra
Under the presumptive test, single strength
(Curcuma longa), Daruharidra (Berberi saristata),
MacConkey broth (5%) was uniformly distributed in
Bruhat Ela (Amomum subulatum), Manjista (Rubia
two sets (containing three tubes each).Whereas,
cordifolia), Bakuchi (Psoralea corylifolia), Rasna
double strength MacConkey broth (5%) was
(Alpinia galanga) and Ushira (Vetiveria zizanioides).
dispensed in a set of five tubes. Water samples (0.1
Vagbhat’s Ksharapakavidhi was followed for
and 1 mL) were inoculated in the sets of single
preparing the formulation of JNY. JNY was stored in
strength MacConkey broth. The tubes containing
sealed glass bottle under cold condition for further
double strength MacConkey broth were inoculated
use.
with 10 mL of the water sample, following which, all
Table 1: Ratio of ingredients the tubes were incubated at what temperature for
Ash Water Prativapa Dravya 24h[1].
Quantity 1 Drona 2000 Pala 8 Pala The tubes showing the positive results were
JNY 100 g 800 g 16 g further analysed by incubating the tubes for 24 h
Following its preparation, JNY was analysed after treatment with JNY. MPN in each case was
for organoleptic properties (colour, odour and calculated by using McCardy’s table. The above test
taste),physical characteristics (weight/mL, refractive was repeated after 1 year.
index, specific gravity, pH, viscosity and total solids), 2. Membrane filtration
organic properties(carbohydrates, reducing sugars,
The untreated and treated water samples
hexose sugars, pentose sugars, non-reducing
were filtered with a membrane filter (0.45 µ). The
polysaccharides, amino acids, proteins, steroids,
filter was then incubated in 100-mL MacConkey
tannins and phenolic compounds) and inorganic
broth at 30˚C for 24 h. Any change of colour of the
properties (calcium, magnesium, sodium, potassium,
broth was the indication of microbial growth.[10]
iron, sulphates, phosphate, chloride, carbonate and

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 Int. J. Ayur. Pharma Research, 2018;6(12):26-33
3. Standard plate count distilled water. After serially diluting the respective
The (treated and untreated) water samples concentrations of the formulation, each dilution (1
were inoculated on soybean casein digest agar and
[11] mL) was added into the wells in each of the plates.
Sabouraud’s dextrose agar.[12] The bacterial and The plates were incubated at 37˚C for 24 h and
fungal plates were incubated at 30 ± 2˚Cand 25 ± 2˚C observed for the zone of inhibition[13].
for 24h and 5days, respectively. Following RESULTS
incubation, the colonies developed on the plates were Analysis of Jala Nirvishikarana Yoga
enumerated. 1. Organoleptic test: The formulation was found to
Antimicrobial activity of Jala Nirvishikarana Yoga be a dark brown, alkaline, thick and viscous liquid
Microbial test cultures (Salmonella abony, with a pleasant odour and a characteristic slight
Staphylococcus aureus, Escherichia coli and Candida bitter taste.
albicans) were inoculated onto soybean casein digest 2. Preliminary test: The formulation was devoid of
agar plates. The antimicrobial activity of JNY was any organic compound and had the presence of
evaluated by using various concentrations of JNY sodium, nitrates, iron and sulphates. The physical
(one drop, three drops and five drops) in 100 mL of characteristics, as analysed, are listed in Table 2.
Table 2: Physical characteristics of JNY
S.No. Tests JNY
1 Weight/mL (mg/mL) 620
2 Refractive index 1.359
3 Specific gravity 1.0978
4 pH 10
5 Viscosity (PaS) 7.27 X 10-4
6 Total solids (g/10 g) 0.460
Optimum dose and period
The optimum dose was analysed by taking into consideration 0.25, 0.5 and 1mL of JNY, wherein it was
found that pH of water sample increased from 7.5 to 10.8 with the increase in concentration of JNY. This pH
change was found to be beyond the permissible limits and made the water more alkaline and conductive in
nature (Table 3).
Table 3: Estimation of optimum dose of Jala Nirvishikarana Yoga
S. No. Parameters Permissible limit Control 0.25 mL JNY 0.5 mL JNY 1 mL JNY
1 pH 6.5–8.5 7.5 9.55 10.5 10.8
2 Conductivity (μΩ/cm) 1000 210 300 420 450
3 Turbidity (NTU) 40 34.8 36.8 39.8 42.5
4 Total hardness (mg/L) 300 32 32 40 32
5 Calcium (mg/L) 75 16 28 28 20
6 Chlorides (mg/L) 250 32 32 44 32
7 Alkalinity (mg/L) 200 36 48 84 120
8 Total dissolved solids (mg/L) 1000 120 122 123 125
Hence, the optimum dose was considered as less than 0.25 mL (or three drops) of JNY. Since none of the
tubes showed a considerable change in the parameters considered, 30 min was considered the optimum period
for administration of JNY (Table 4).
Table 4: Estimation of optimum period of Jala Nirvishikarana Yoga
Sl. No. Parameters Permissible limit Control 30 min 1 h 2h
1 pH 6.5–8.5 7.5 7.99 7.99 7.99
2 Conductivity (μΩ/cm) 1000 210 300 302 300
3 Turbidity (NTU) 40 34.8 36.8 36.9 37.1
4 Total hardness (mg/L) 300 32 32 33 32
5 Calcium (mg/L) 75 16 28 29 29
6 Chlorides (mg/L) 250 32 32 31 31
7 Alkalinity (mg/L) 200 36 48 48 48
8 Total dissolved solids (mg/L) 1000 120 122 128 132
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 Shivanand B. Kalasannavar et al. Efficacy of Vagbhatokta Jala Nirvishikarana Yoga on Polluted Water
Stability test
The colour remained unchanged even after 1 remained the same till 1 year of storage. pH of JNY
year of storage in the case of both the samples. The was constant (7.99) for approximately 3 months and
taste and odour was found to slightly decrease in then decreased to pH 9.0 for both the samples.
quality after 4 days of storage but thereafter
Microbial limit test
1. MPN
The count of coliforms in the river water sample (100 mL) was found to be 180, which reduced
considerably to 8 after treatment with JNY. (Table 5, Figures 1& 2).
Table 5: Analysis of treatment by MPN method
Samples Before treatment After treatment
River water 180 MPN 8 MPN
Well water <1 MPN -
Borewell water <1 MPN -
Tap water <1 MPN -
2. Membrane filtration
Prior to the treatment of water sample with JNY, the sample showed the presence of faecal coliforms per
250 mL (yellow-coloured broth) indicating the coliform contamination. However, the broth inoculated with
the membrane filter of the treated water sample displayed no change of colour indicating inhibition in the
proliferation of coliform by the administration of JNY.
3. Standard plate count
The untreated water sample showed the presence of 267 CFU/mL of bacterial colonies on soybean casein
digest agar and 98 CFU/mL of fungal colonies on Sabouraud’s dextrose agar. Conversely, upon treatment
with JNY, the bacterial and fungal counts were reduced to 83 CFU/mL and 15 CFU/mL, respectively. (Table
6, Figure 3).
Table 6: Analysis of effectiveness of JNY by Standard Plate Count method
Bacterial count Cfu/ml) Fungal count (Cfu/ml)
Sample
2012 2013 2012 2013
River water 267 83 76 53 98 15 45 31
Well water 28 - 35 - 05 - 06 -
Borewell water 32 - 40 - 12 - 11 -
Tap water 105 - 29 - 29 - 08 -
*BT, Before treatment; AT, After treatment
Antibacterial activity
All the concentrations of JNY (one drop, three drops and five drops in 100 mL of distilled water) were
found to be effective in inhibiting the growth of the organisms (Salmonella abony, Staphylococcus aureus,
Escherichia coli and Candida albicans).

Figure 1: River water-color change in Double Standard McConkey

Broth & Standard McConkey Broth tubes
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 Int. J. Ayur. Pharma Research, 2018;6(12):26-33

Figure 2: River water + JNY- No change in Double Standard McConkey Broth & Standard McConkey
Broth tubes

Figure 3: Results of inoculated plates showing no growth

DISCUSSION
The study was conducted to assess the sodium, nitrates and iron are contributed by ash of
physiochemical and antimicrobial efficacy of JNY on the drugs.
Malaprabha river water, which has been reported to Optimum period of administration of JNY was
be have high pH and Biological Oxygen Demand kept to minimum (30 min) even though no significant
(BOD) levels, in the recent years[14, 15]. As a river can change in the parameters was observed with
be a used for various activities (e.g. irrigation, prolonged incubation period. Among the parameters
washing and drinking), the elevated level of BOD may analysed for determining the optimum dose of JNY,
adversely affect the health of inhabitants living near significant variation was detected in pH of the water
the banks of the river. Hence, the traditional method probably due to the alkaline nature of JNY. Hence,
of treatment has been attempted to remedy the least dose of JNY (approximately three drops in 100
problem of pollution by heavy metals and mL) was considered as the optimum dose to
microorganisms. minimise the change in pH. Moreover, at such a low
JNY, the Ayurvedic formulation, was found to volume, the odour and taste were appreciable, which
be alkaline and viscous in nature, which may be due further helped to ascertain the optimum dose. The
to the presence of Kshara and Sarala Niryasa. Kshara characteristic odour may have developed due to the
can lead to increase in the pH of the formulation, volatile compounds present in Bruhat Ela.[18]
thereby giving rise to its alkaline nature[16] whereas Ushira[19] and Bakuchi.[20] However, for highly
the viscosity may be as a result of the dense and polluted water, the dosage of JNY should be increased
resinous nature of Sarala Niryasa.[17] The complete accordingly, as low concentrations of JNY may not be
oxidation of herbs upon burning resulted in the sufficient to reduce the microbial load of the water.
absence of organic component in JNY. However, the The water sources often vary in their physical
inorganic constituents in JNY such as sulphur, properties, especially in pH, due to the presence of

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 Shivanand B. Kalasannavar et al. Efficacy of Vagbhatokta Jala Nirvishikarana Yoga on Polluted Water
several organic and inorganic substances. The may result in improved activity of the formulation.
sources of water with low pH may contain higher Ativisha being a costly herb, more cost-efficient herbs
concentrations of toxic metals[21], which can become can be tested and used to make the formulation more
detrimental to the health of the consumers. affordable.
Malaprabha river has been reported to contain iron CONCLUSION
beyond permissible levels, which may be as a result JNY, a novel Ayurvedic formulation prepared
of leaching from rocks due to low pH of the river[14,15]. in the present study was observed to have substantial
JNY, due to its alkaline nature, can therefore be used antimicrobial activity with the ability to limit the
to maintain the pH of the water to some extent growth of coliforms, Salmonella abony,
leading to decrease in the leaching of metals and Staphylococcus aureus and Candida albicans. This
limiting the levels of BOD. property could help in enabling the purification of
Although there was no substantial change in water before consumption, especially at the
the taste and odour of the formulation, significant household level. In addition, the formulation was
post-storage change was observed in case of pH of found to be stable for 1 year at room temperature.
JNY. Kshara contributes to the increase of pH of JNY can therefore be stored and used for a long
JNY[16] and has been stated to possess antibacterial period of time. However, further research is required
activity against E. coli and S. aureus[22]. Small et al.[23] to enhance the dose concentration and effectiveness
has also stated that the survival rate of E. coli of JNY. Uniformity of the effectiveness of JNY should
(aerobic log-phase cultures) was affected as a result be further assessed by including samples from
of increase in the pH of the medium. Hence, it can be different water sources.
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*Address for correspondence

Cite this article as:
Dr Shivanand B. Kalasannavar
Shivanand B. Kalasannavar, S. K. Hiremath, U. S. Hampannavar. An
Experimental Study on the Efficacy of Vagbhatokta Jala Nirvishikarana Yoga Assistant Professor,
on Polluted Water. International Journal of Ayurveda and Pharma Research. Department of Agada Tantra, Govt.
2018;6(12):26-33. Ayurveda Medical College, Mysure,
Source of support: Nil, Conflict of interest: None Declared Karnataka, India.
Email: [email protected]
Tel: +91994509149
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