1.

INTRODUCTION

The name of product is spelt as GAUNYLE. It is a floor cleaner that has been made

by tapping the microbial properties of Desi Govansh urine and neem. The formula has

been developed by the Ministry of small & medium enterprises (MSME)., Govt. of India,

Bioprocessing & Herbal division MahatmaGandhi Institute for rural industrialization

(MGIRI), Wardha. The name is registered as a property of Holy cow foundation. Patanjali

intention to protect the Desi Govansh and at the same time the product being hold by

patanjali has nothing to do with us except for the deceptive similarity in the name. The

name is registered in the name of Holy cow Foundation since 2015.(The good will project

Jan.23,2017)

1.1 GAUNYLE

Gaunyle is a floor cleaner and has Gaumutra as a component and the term

phenyl is commonly used for floor cleaners, the presence of the cow element,

and also to denote its function of cleaning floors. Chemical analysis of desi

1
govansh urine and govar proves that it has powerful microbicidal properties.

The uniqueness of the desi govansh is recognized by Ayurvrda in ancient times.

Goumutra is boiled for a long time during the process of making Gaunyle.

Gaunyle. Gomutra refers to the usage of cow urine for therapeutic purposes in traditional

Indian medicine, Ayurveda. It is also an important component of the mixture called

Panchagavya also used in Ayurveda.(T V Sairam 16 january 2008). Cow is considered special

and it contain special hormones and minerals. It is also used in Vaastu shastra for

purification purposes. Sprinkling of cow urine is said to have a spiritual cleansing effects

as well.(Kamadhenu Sutra 10 march2015). Cow urine is helpful in the treatment of

leprosy

and cancer. It is also used in the treatment of fever by mixing it with black pepper,

yoghurt,, and ghee. A mixture of gomutra, neem bark, vasaka bark, kurilo bark, kaner

leaves. In study from Mandsaur has claimed that it may also benefit cancer patients.

Cow urine is also used in Myanmar and Nigeria as a folk medicine.(N.K. jain .,et al 2010-

2
2015)

In 2002, a US patent is issued to a group of Indian scientist from Council of

Scientific and Industrial Research (CSIR). The cow urine was claimed to be serving as a

bioenhancer, enhancing anti-microbial activity of antibiotic and antifungal agents. It is an

activity of anti-microbial & agents.(Indian patents cow urine for medicinal use, july 2002)

Micro-organism (bacteria & fungi) are mainly responsible for the increased in human being

and also animals, as these create problems in treatment.

1.2 COW URINE

Cow urine and all Indian systems of medicine has a strong scientific base. The

cow is proved to be beneficial in the areas of agriculture, science and technology, energy,

industry, medicine, etc for the development of any community, in eco-friendly in nature. In

the present study the antibacterial potential of cow urine were investigated.

The Indian cow Bos indices, is the most valuable animals in religious scriptures. Cow

3
urine has found therapeutic applications since days. Cow urine is consumed by the

majority of the rural population as the traditional medicine in almost the whole Indian

continent.

Cow urine is based preparations is able to the viral, microbial, and fungal pathogenic

conditions. These are powerful antimicrobial, antiviral , anti-allergic and anti-oxidant

activity.

India has a large geographical area with different culture practices that are medicinal plants.

The Indian Ayurveda is the result of the medicine developed by means of documenting over

25000 herbs.

1.3 TRADITIONAL USES OF COW URINE

In India the rural villages, cows urine is used as a very long time for the antiseptic for the

wounds, skin diseases, bathing etc. cow urine is used in many drugs formulations and also used

in purification. Around 5 years old it is proved that cow urine is most effective natural antiseptic

and disinfectant in comparision to the synthetic chemicals that are recently available to the

consumers.

It is not a toxic effluent like 95% is content of water, 2.5% is urea and remaining mixture of

4
mineral, salt, harmones and enzymes is 2.5%. Ancient time Indian vedic scriptures including

Manu smriti, charaka Samhita and this time researchers of used this animals product separate any

non-functional of respiratory systems, cardiovascular system, cancer and so many other.

1.4 TRADITIONAL USES OF COW URINE AND DRUGS USED WITH

COW URINE

DISEASES CONSTITUENTS OF GOMUTRA ALONG

WITH ADJUVANT
Fever Urine,pepper,curd ghee
Leprosy Dhruhardi , urine
Deformation condition in leprosy Nimbuchal , urine
Chronic leprosy Vasaka leaves, kuraila bark, kaner leaves,
neem bark, urine
Epilepsy Neem bark, somapada bark, mustard oil,
urine
Anemia a)Cow milk,urine,tripala
b)Loh bhasma, urine, milk

5
1.5 COW URINE

Cow urine therapy destroys all the pathogenic organisms and it is used in daily routein, it boosts

immunity. It is proved that some diseases are cured by cow urine such as cough, migraine or

headache, itching, acne, heart disease,

AIDS, diabetes mellitus, blood disorders, urological disorder, asthma, kidney shrinkage, hepatic

disorder, cancer etc. cow urine is also help in maintaining the homeostasis of body where it

affects body function of lowering cholesterol level, relieving tension, improving memory,

functioning of liver, decreasing the aging process, destroy the toxic effects of medicinal level in

the body.

Cow urine is regularly used without having any illness, our body is healthy by the boosting

immunity, by separating toxic substance by generation of antioxidants and free radicals.

Recently it proved that cow urine is the potential source of natural antioxidant. It important that

preventing of supportive the therapy and decreasing the stress related degenerative diseases.

(^a b India today5 May 2015)

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1.6 COW URINE USED IN AGRICULTURE

Cows urine boosted the annual rye grass break down by causing an increase in nitrogen (N)

component of the soil and depression in N fixation by 10% annually in clovers particularly in

winter. Urine increased the N concentration of grass ( particularly the nitrate fraction) and

increased the potassium concentration of grass and clover.

Total N concentration in the cow urine is very high ranging from 6.8 to 21.6 g N/I., average off

69% is urea. Then the effects on yield lasted 2-3 harvests and decreases in clover growth.

1.7 ANTIBACTERIAL ACTIVITY

Antibacterial activity are the most important tools in fighting bacterial infections and it is

benefit for health related quality of human life.

Anything that destroys bacteria or suppresses their growth or their ability to reproduce.

Many antibacterial products for cleaning and handwashing are sold. Such products do not

reduce the risk for symptoms of viral infectious diseases. The potential contribution of

antibacterial products to reducing symptoms of bacterial diseases in the home.

7
Any how these benefits are under treated as many commonly used antibiotics have become less

and effective against certain illnesses not, because so many produces toxic reactions but also

due to emergence of drug- resistance bacteria.

Drugs are derived from natural sources play important role in the prevention and treatment of

human diseases. The major role of bacteria have genetic ability to transmit and obtain resistance

towards the drugs and there are also adverse effects of drugs on the host.

Cow is the most valuable animal and it is called mother of all. Different products obtains from

cow such as urine, drug, ghee, milk, curd, and so many numbers of Ayurvedic formula of the

medicine. Cow urine are many medicinal properties and use for curing numbers of diseases such

as skin diseases, kidney, anemia, constipation, respiratory diseases etc.

1.8 USEFUL CONTENT IN COW URINE

1-UREA: Urea is a major found in urine and is the end product of protein

metabolism. It is strong antibacterial agent.

2-URIC ACID: Uric acid is similar to urea and has strong antibacterial properties. In

addition, it help to control cancer-causing substances.

3-MINERALS: Minerals from urine can be very easily reabsorbed as compared to those

8
 derived from food. Urine mostly contains different types of minerals then

those are obtained from food.

Urine becomes turbid if left alone for a white. This is because when enzyme present

in urine dissolve urea and change it into ammonia then urine becomes strongly

alkaline making it difficult rich minerals.

Therefore urine looks turbid ; this does not means that it has decayed urine with a

higher ammonical disorder content when applied to the skin plays as an important

role in beautifying it.

9
2. LITERATURE REVIEW

The current research is focussed to evaluate the comparative intro antimicrobial

activity of cow urine with medical plant extracts agains common pathogenic

microbial strains. The detail study of antimicrobial potential of (i) fresh cow

urine, (ii) cow urine distillate, (iii) sterile cow urine on pathogens like Bacillus

subtilis, Escherichia coli, Salmonella typhi, Proteus vulgaris. It indicates that cow

urine and medicinal plant extracts exhibit better antimicrobial action against

different clinical microbial strains hence it can be utilized to control microbial

infections. (Farida p Minocherenhomji ; oct-dec., 2014)

The use of chemicals of control infection due to micro-organism, more problems

have been created rather than being addressed. Disinfectant prepared during this

study is 100% natural and is based on cow urine and biodegradable and

antibacterial plant extracts. ( Neem, Tulsi, Ritha, and Pine oil ) possessing pleasant

aromatherapeutic vapour.

No additional artificial dyes or fragrances have been used. The result showed that

10
the higher concentrated of both Neem and Tulsi extract possess maximum efficacy.

The main aim of this product development is to provide employment to rural youth

and to use safe disinfectant for cleaning floors, etc. ( S A Mandavgane,et.al; 2005)

Gaunyle is a disinfectant obtained from cows urine . The name Gaunyle is

registered as a property of Holy cow foundation. This name is registered in the

name of holy cow foundation since 2015.

Gaunyle is a floor cleaner and has Gaumutra as component and term phenyl is

commonly used for floor cleaner, gaumutra is boiled for a long time during the

process of making Gaunyle. Many other products are manufactured using

distilled Gomutra. Ayurvedic aapplications to make traditional as well as modern

Panchagavya based products like face packs, hair oil, shampoo, joint pain oil,

Gaunyle and more.(The good will products , 25 march 2017)

Anuradha modi, heads the holy cow foundation, said Gaunyle has the anti-

microbial and anti-fungal properties of cow urine and neem. The products is

being sourced from cow sheds in Mathura district of U.P. Initially the products

it has a strong smell. Nobody would have used it. So we have distilled the

urine now and added natural intgredients like pine oil to cover the smell.

(Pravin singh.,2015)

11
Gaunyle with the anti-microbial and biodegradable properties of cow urine

shampoo with gomaya and gomutra. An economic times report says gaunyle soon

replace the commonly used phenyl to clean government offices and will be

shocked at centrally run chain of Kendriya Bhandras.

In India cow is equated to the mother and her urine is helpful for all the heart attack,

piles, arthritis, asthma, cancer, acidity, constipation, ulcer, blockage in arteries,

blood pressure, diabetes, migraine, thyroid, eczema, gynaecological; and also used as

increase the nitrogen content of the soil, better rearing of honeybee.

In natural, eco-friendly with no side effects available, therefore it can be of cow urine

distillate was analyzed against the Bacillus subtilis, Pseudomonas aeruginosa,

Salmonella typhi. It conclude that, the cow urine has antibacterial activities and

inhibitory activity can be used in the control of bacteria of various origin.

Mostly these products provides to synthesize new structural types of antimicrobial

chemicals that are safe to man. Crude extracts of some well medicine plants are used

to control plant pathogens. The antibacterial activity of methanol extracts and

petroleum ether, chloroform and ethyl acetate fraction from the root bark of Akanda

(Calotropis gigantea) were investigated by Ashreful et al., 2008.

12
Indiscriminate uses of antibiotics have caused bacterial resistance and also lead to

many side effects. In some cases plants and animal materials are widely used for the

treating various ailments due its antibacterial products. In Ayurveda, cow urine has

been used to general health of an individual. The method employed tostudy of

antibacterial activity is agar well diffusion technique. The result showed good

antibacterial activity of cow urine against most of the test bacterial strains by

exhibiting zone of inhibition.

In urine is usually support the growth of non bacteria UTIs(Cattell etal.,1974). Urine

from normal individuals may be inhibitory , especially when the inoculums is small

(Kaye,1968). The most inhibitory factors are the osmolality, urea concentration,

organic acid concentration, and pH. Bacterial growth is inhibited either by very

dilute urine or a high osmolality when associated with a low pH. Much of the

antibacterial activity of urine is related to a high urea and content (Solomon

et.al.,1983). From a clinical perspective, anyhow conditions do not ppear to

13
 significantly distinguish between patients are susceptible or resistance to infection.

In the scenario about 70% of pathogenic bacteria are resistance to the least one

of the drugs for the treatment, is to be taken from traditional medicine to its

urgently. The Indian traditional knowledge from ayurveda,where Bos indicus is

placed at a high pedestal for numerous uses of its various products. Urine is one

of the products of a cow with many benefits and without toxicity. Interestingly

antibacterial activity has also been found against some resistance strains.

Antibacterial action is still further by it being an immune-enhancer and

bioenhancer of some antibiotic drugs. The cow urine and its distillate tested for

antioxidant and antibacterial activity exhibited the mentioned activities and

comparatively fresh cow urine was found to the better than its distillate(N.K. Jain;

2015)

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3.1 EQUIPMENT AND THEIR USES

1- ORBITAL SHAKING INCUBATOR

Incubator is a device used to grow and maintain microbiological culture or cell culture.

The incubator maintain optimal temperature, humidity and other condition such as the

carbon dioxide and oxygen content of the atmosphere inside. The most commonly used

temperature both foe bacteria such as requently used s.cerevisiae is approximately 30 c

as these organism grow well under such condition.

Fig.1- Orbital shaking incubator

15
2- LAMINAR AIR FLOW- A laminar flow cabinate is a carefully bench

designed to prevent contamination of semiconductor wafer, biological sample or

any particle sensitive material. Air drown through a HEPA filter and blow in a

very smooth, laminar flow towards the user. The cabinate usually made of

stainless with no gap or joints where spores might collects. Laminar air flow

cabinate may have a UV-C germinate lamp to sterilize the interior and content

before usages to prevent contamination of experiment.

Fig.2- Laminar air flow

16
3- MICROCENTRIFUGE- A laboratory centrifuge is a piece of driven

by a motor, which spins liquid samples at high speed. There are various types of

centrifuges, depending on the size and the sample capacity. The centripetal

acceleration is used to separate substances of greater and lesser density.

Fig.3- Microcentrifuge

17
4- AUTOCLAVE- An autoclave is a pressure chamber used to carry out

industrial process requiring elevated temperature and pressure

different from ambient air pressure. The autoclave carries out exact

function of sterilizing materials. Micro-organism are bacteria, virus,

fungi, parasites etc, they are able to cause infection in our body.

Spores are the environment resistant form of the micro-organism, they

can be killed if extreme condition are maintained for an extened

period of time.

18
 Fig.4- Autoclave

5- ELECTRIC BALANCE- Electronic balance is to measure weight

or calculate mass spring balance or spring scale measure weight

(force) by balancing the force gravity against the force spring , where

balance or pair of scale using balance due to the mass of an object

against the own mass or masses. Either type can be calibrated to read

in unit of force such as newton or in unit of mass such as kilogram,

but the balance or pair of scale using a traditional balance being to

compare masses will read correctly in force in a different gravitational

field strength, while the spring balance would read correctly in force

in a different gravitational strength.

19
 Fig.5- Electronic balance

6- MAGNETIC STIRRER- It employs a rotating magnetic field to cause a stir

bar(also called flea) immersed in a liquid to spin very quickly, thus stirring it.

The rotating field may be created either by a rotating magnet or set of stationary

electromagnet, placed beneath the vessel with the liquid.

A stir bar mixing a solution on a combined hot-plate magnetic-stirrer device. The

20
left knob controls the stirring rate and the right knob controls heating.

Fig.6- Magnetic stirrer

21
3.2 GLASSWARE AND OTHER MATERIALS

USED

S.No Name Objective

1 Conical flasks As a container of media & culture flask
2 Inoculating loop To inoculate the fungal disk into media
Pipette (20, 50,100, 500,
3 For pipetting
750, 1000 l)
4 Measuring cylinder To measure the volume of reagent

5 Volumetric flask To maintain the volume of buffer,substrate etc

6 Petriplates Culture growth, Inoculum preparation etc

3.3 MATERIALS AND METHODS

3.1 MATERIAL REQUIRED

Autoclave

Heating metal

Double distilled water

Incubator

Nutrient agar

Laminar air flow

22
3.2 SAMPLE OF GAUNYLE

Fig.7

Sample 1 of # 1 GauGram Gaunyle

Sample 1 of # 2 GauGram Gaunyle
Sample 1 of # 3 GauGram Gaunyle
Sample 1 of # 4 GauGram Gaunyle
Sample 1 of 3 5 GauGram Gaunyle

23
3.3 GROWTH MEDIA

Culture media is a solid, liquid, semi-solid designed to support the growth of

microorganism or cells . Media are used for growing different cells.

The most common growth media for microorganisms is nutrient agar plates, specialized

media are sometimes required for cell culture growth. Some organisms, termed fastidious

organisms require specialized environment due to complex nutritional requirements.

3.4 PREPARATION OF NUTRIENT AGAR

* Distilled water

* Measuring cylinder

* Laboratory spoon

* cotton gloves

* Sterile, empty petri plates

* Autoclave

*Incubator

The component of Nutrient agar media for bacteria is---

Nutrient Agar- 28g

Distilled water- 1000ml

24
3.5 PREPARE A MEDIA

1- Media component is required amount were added into a conical flask containing doubled
distilled water.

2- The mouth of flask was plugged with cotton and covered with paper wrapping around it.

3- Then, kept them into autoclaved for at 121 c for 15 minutes.

25
 Fig.8- Nutrient agar media

4- After autoclaving the flask cooled at room temperature. Then, kept in the laminar flow
unit under UV radiation for 20 minutes.

5- After 20 minutes media poured each petriplates to volume.

6- Label the plates according to the medium and perform a sterility test as in.

7- In case of longer storage, petri plates must be placed into plastic bags or boxes to avoid

26
 drying out.

Fig.9- Pouring plate

8- Allow plates to cool until solid and streaked bacteria onto the media

Fig.10- Striking plate

27
9-Transfer plates to the cool untill solid and steaked bcateria onto the media incubator at
30c for 48 hrs. .

Fig.11- Striking the media plate with microorganism

Make the three drill in media plate, put the sample 1 of # 1, the concentration is
20 l.

In second media plate make the drill and put the sample 1of # 2, the
concentration is 20 l.

Then the third media plate make the three drill and put the sample 1 of # 3,
28
 the concentration is 20 l.

In fourth media plate make the drill and put the sample 1 of # 4, the concentration
is 20 l.

And the last media plate make the three drill and put the sample 1 of # 5, the
concentration is 20 l.

Fig.12 Media of samples

29
 Fig.13- Largest zone of inhibition occur in sample 1 of # 5

SEM PROCEDURE FOR BACTERIA

In laminar chamber take a culture of bacteria with the help of loop.

A small quantity of bacteria put in appendrof tube, add 2 ml of distilled water

and shake it well then centrifuged till 5000 rpm for 5 minutes.

After centrifugation discard distilled water from tube and 30% ethanol in tube.

In appendrof tube (bacteria) put in incubator at 30c for 10 minutes, after

30
 incubate; centrifuge for 5 minutes at 5000 rpm.

Discard 30% ethanol from tube and add 50% , 70%, 90% ethanol in tube

respectively

100% ethanol add in tubes then left for 10 minutes then after One drop of

sample of bacteria put on slide and cover with coverslip.

31
Fig.14- SEM (Scanning electron microscope) for antibacterial analysis of Gaunyle

``````````````````````````

32
RESULT AND DISCUSSION

Sample Bactria Sample Zone of Growth

Name amount inhibitio Pattern
n (cm)
Sample-1 20l to 1.2 Effective
6000l at 5000
l
Note:
growth
observed
Sample-2 20l to 1.5 Effective
6000l at 5000
l
Note:
growth
observed
Sample-3 20l to 1 Effective
6000l at 5000
l
Note:
growth
observed
Sample- 20l to 0.5 Effective
4 6000l at 5000
(Without l
GM) Note:
growth
observed
Sample-5 20l to 2.5 Effective
6000l at 5000
l
Note:
growth
observed
Pine 20l to 0 Growth
6000l observed
at all
tested
33
concentra
tion
 Tulsi 20l to 0 Growth
6000l observed
at all
tested
concentra
tion
Neem 20l to 0 Growth
6000l observed
at all
tested
concentra
tion
Commer 20l to No zone No
cial 6000l sustainab
Phenyl Growth ility of
from microorg
20l to anisms
900l
at 1000
Effective l
at 1000

TESTING REPORT OF GAUNYLE

Results: (1) Sample -1, 2, 3, and 5 has antimicrobial activity as inhibition growth was observed.

(2) The antimicrobial activity is not very effective as sufficient growth was observed at

fix concentrations.

(3) Sample-4 has realizable antimicrobial property which cannot be considered.

34
The antibacterial activity of the study is also constituents with the reports .Other authors

which show good antibacterial activity in fresh urine. Urine distillate was not found to be

active among all the cow urine preparations. It may also be due to presence of more

cations and formation of nitrosoaamines.

CONCLUSION

The result indicates that the tested Gaunyle have strong antimicrobial activity and should

zone of inhibition on nutrient agar plate. Finally, it was concluded that highest zone

inhibition 2.5 was observed in sample 5.

As cow urine show that antibacterial activity against the pathogenic bacteria, which is

selected for further to isolate bioactive natural constituents that may needs for therapeutic.

But, in vivo studies on these cow urine preparations are necessary and should seek to

justify its potential as antimicrobial agents.In research in this direction with the help of

develop proper extracts to counter various microbial infections, and boosting the human

immune system.

It is the most effective natural remedy and the safest method of treatment gave upon by

the nature. The constituents of the cow urine and its medicinal important and has brought

in different phases of life.

35
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