PREPARATION OF NATURAL MOSQUITO

REPELLENT

Introduction:

A. indica belongs to Meliaceae, a family of dicots mostly represented by trees

and shrubs.The family includes about 51 genera and 550 species, with many of
them prized for their wood, edible fruits, and medicinal and ornamental
qualities (Wiart, 2006). It is a small to medium-sized evergreen tree with a
height of 15 m (30 m maximum), having a larger rounded crown (10–20 m)
with spreading branches and a branchless bole (7.5 m, diameter90 cm). The
bark of the tree is thick, fissured, dark gray to red (inside) in colour, and it
possesses a gummy colourless sap. The leaves are long (20–40 cm), alternate,
pinnate,exstipulate, and glabrous with a light green hue. The leaves have two
pairs of basal glands [1]
With a subglabrous petiole (2–7 cm) and above, channeled rachis. Each leaf
comprises8–19 serrated, proximally alternate, ovate to lanceolate leaflets.
Inflorescence is axillary clustered multiflowered thyrsus (150–250 flowers) with
a length of 15–30 cm and minute caducous bracts. Flowers of the tree are small
(1 cm in diameter), white or pale yellow, and sweet smelling.They are
actinomorphic, pentamerous, and bisexual or unisexual maleon the same plant.
The calyx of the flowers is imbricate, ovate, thin, and puberulous from

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Inside, while petals are free, spreading, imbricate, spathulate and ciliolate from
inside.Fruits are single (maximum of two) and small (1–2 cm) in size. They are
greenish to yellow in colour and an ellipsoidal seeded drupe. The tree has a thin
exocarp, pulpy mesocarp,[1]

Chemical constituents:

Neem compound Source Biological activity Reference Nimbin Seed oil

Spermicidal (Sharma, 1959) Nimbolide Seed oil Antibacterial Antimalarial
(Rochanakij, 1985), (Khalid, 1989), (Rojanapo, 1985) Gedunin Seed oil
Antifungal Anti malarial (Khalid, 1989), (Rao, 1977) Azadirachtin Seed Anti
malarial (Jones, 1994) Mahmoodin Seed oil Antibacterial (Devakumar, 1996)
Gallic acid, Epicatechin, Catechin Bark Anti-inflammatory and
immunomodulatory (Van der Nat, 1991) Margolone, Margolonone and
Isomargolonone Bark Antibacterial (Iffat, 1989) Cyclic trisulphide and Cyclic
tetrasulphide Leaf Antifungal (Pant, 1986) [1-10]

Uses:

1. Antibacterial activity Recent research shows the isolation and identification

of the antibacterial active compound from petroleum ether extract of neem
oil (Zhang, 2010). Elavarasu et al. studied in vitro anti-plaque microbial
activity of neem oil (Elavarasu, 2012)

2. Antiviral Galhardi et al. studied the in vitro antiviral property of Azadirachta

indica polysaccharides for poliovirus (Galhardi, 2012). The research of Xu et
al. showed the in vitro antiviral activity of neem seed kernel extracts against
duck plague virus (Xu et al, 2012).

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3. Sexually transmitted disease Few researchers have focused on neem efficacy
in treating sexually transmitted diseases. The reports that have been
completed are overwhelmingly positive. Recent research of Shokeen et al.
showed the evaluation of the activity of 16 medicinal plants against
Neisseria gonorrhoeae (Shokeen, 2009). National Conference on “Recent
Advances in Zoology” (NCRAIZ-2018) www.ijlsci.in Int. J. of Life
Sciences, Special Issue A10; January, 2018 | 183

4. Neem and the immune system Thoh et al. studied that azadirachtin interacts
with the tumor necrosis factor (TNF) binding domain of its receptors and
inhibits TNF induced biological responses (Thoh, 2010).
5. Anti-inflammatory activity The study of Alam et al. showed the anti-
inflammatory activity of epoxyazadiradione against macrophage migration
inhibitory factor (Alam, 2012). Thoh et al. found that azadirachtin interacts
with retinoic acid receptors and inhibits retinoic acid-mediated biological
responses (Thoh, 2011).

6. Antioxidant effect Manikandan et al. researched that antioxidant and

protective effects of active neem leaf fractions against hydrogen peroxide
induced oxidative damage to pBR322 DNA and red blood cells
(Manikandan, 2009).

7. Anticarcinogenic activity Chatterjee et al. showed that identification of a

sulfon oquinovosyldiacylglyceride from A. indica and studies on its
cytotoxic activity and DNA binding properties (Chatterjee, 2010). Mahapatra
et al. showed novel molecular targets of Azadirachta indica associated with
inhibition of tumor growth in prostate cancer (Mahapatra, 2011).

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8. Skin diseases Neem has a remarkable effect on chronic skin conditions.
Acne, psoriasis, eczema, ringworm and even stubborn warts are among the
conditions that can clear up easily when high quality, organic neem oil is
used. Neem oil and leaves has been used in Siddha medicine for the
treatment of skin diseases (Thas, 2008). In addition, neem oil can be used as
an excellent component of cosmetics to help clear, beautify and rejuvenate
the skin.

9. Antisnake venom activity Ashis et al. studied a snake venom phospholipase

A2 (PLA2) inhibitor (AIPLAI) was isolated from leaves of A. indica (neem)
and the mechanism of PLA2 inhibition by AIPLAI in vitro condition was
also studied (Ashis, 2008).

10.Digestive disorders Neem is generally accepted in the ayurvedic medical

tradition as a therapy for ulcers and other types of gastric discomfort. Neem
promotes a healthy digestive system by protecting the stomach, aiding in
elimination [11-17]

Markteted available Products

Protecting oneself against mosquito bites is a big deal and matter of concern in
India. Mosquito Bites can cause infections and lead to a serious disease called
malaria. It is necessary to use a mosquito repellent coil or cream or liquid to
protect our body against those infections. Here is a list of the best Mosquito
Repellents in India :

 Good Knight Mosquito Repellent Liquid

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Good Knight Mosquito Repellent is one of the most popular Mosquito
Repellents of India. The brand is in the market for several years and is rated
high by users. It effectively protects against mosquito bite transmitting
infections like malaria, chickengunya and dengue.

 All Out Mosquito Repellent Liquid

There are so many places in India, which are seriously affected by mosquitoes
and insects and annually a number of people die due to the diseases transmitted
by mosquitoes and other insects. Since its inception in the 90’s of the last
century, All Out has been a trustworthy insect killer for the household purposes.
Not only sprays, but All Out also has a wide range of liquid vaporizer to make
your sleep comfortable at night without any disturbances from mosquitoes and

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insects. Extensive research showed that not only in India, but also all over the
world, people are more or less affected with the torture of insects. There are
some places in India where the disturbances of the mosquitoes are too much and
which causes the death of so many people annually. All Out has become a
stalwart in the industry for more than two decades.

 Mortein Mosquito Repellent Liquid

Mortein is a renowned name in household insecticide. It is basically an

Australian company and is a very popular insect repellent. Mortein has made a
significant place in Australian household. Mortein has the most popular slogan
of “More Smart, more safe, Mortein.” The word Mortein is made by the
combination of two words “mort” meaning (dead) and “ein” meaning (one). The
main active ingredients used in Mortein are Allethrin, Pyrethroid and
Resmethrin. All these are neuro toxins which can be harmful to humans when
taken in typical doses.

 Odomos Mosquito Repellent Cream

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Odomos is a clinically proven brand of mosquito repellent that offers the most
effective protection from mosquitoes, the carriers of deadly viruses that cause
diseases like Dengue, Chikungunya, Malaria, Filaria and Encephalitis etc.

Independent institutes have clinically tested and certified Odomos to be the best
protector against mosquito bites for as long as 12 hours.

 Good Knight Mosquito Repellent Cream

Good knight Naturals Mosquito Repellent Cream is a unique blend of Active &
Exotic natural ingredients, which not only provides complete protection from
mosquitoes but is also safe on skin. Enriched with Tulsi & Lavender, it contains

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 Milk Protein Pearls that moisturise the skin and make it soft.

 Himalaya Herbal Mosquito Guard

Himalaya Herbals Mosquito Guard is a herbal solution to protection against

mosquito bite transmitting diseases, the lotion has protective natural elements
which prevent infections like malaria and dengue. This has proven to be on eof
the best anti mosquito bite aolutions in the country, where infections from
mosquitoes is a common problem.[11-18]

Chemic
als use in synthetic mosquito repellent:

 DEET. DEET, which is an acronym for N,N-Diethyl-meta-toluamide, is one of

the most effective bug repellents and also repels ticks.
 Cyfluthrin. Cyfluthrin structurally resembles DDT and has a similar mode of
action. ...
 Permethrin. ...
 Pyrethroids.[19-24]

Side effect of synthetic mosquito repellent:

Most common reported side effects are eye irritation, headache,

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frequent allergic rhinitis and difficulty in breathing especially for asthmatic
patients.
The side effects are more commonly seen in young children and babies. Hence,
they should be kept away from such repellents as much as possible.
They Are Free Of Dangerous DEET [25-28]

The Benefits of Using Natural Organic Repellents:

N, N-Diethyl-3-methylbenzamide is also known as DEET which is common in

most standard insect repellents offers protection to various insects like ticks,
fleas and mosquitoes. This ingredient is harmful to human’s central nervous
system.  If it happens to mix with other repellent chemicals, it will turn more
toxic and even more dangerous.

They Are Friendly and Kind on Sensitive Skin

Majority of insect repellents contain substances like petroleum and other

synthetic chemicals that may cause skin irritations and allergic reactions. Many
people can use natural organic repellents even in the presence of children, as
they are free of such harmful chemicals.

They Are Good For the Human Body

The essential oils used in these natural repellents have a pleasant smell. In
addition to that, they are known to be beneficial to the human body, for
instance, peppermint oil which can improve an individual’s concentration and
make the muscles relax.

The Combination of Various Natural Organic Repellents Work Better

When you combine different natural insect repellents, they are considered to
work better than any DEET repellents.

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On that note, at DA Aromatherapy Collection, you can choose from a range of
our natural organic repellents, and you will not miss a Natural Tick and
Mosquito Repellent.

Reason for using neem as mosquito repellent:

The neem leaf is also wonderfully soothing for skin that is irritated from the
heat and insects. But realistically these sprays do not offer quite the protection
that you would get from a neem oil/coconut oil mix.

Well, it depends what species of mosquito you are dealing with. For malaria
protection neem oil is fantastic. If you combine the 96-100% protection rate of
the home made mosquito repellent with burning some neem oil when sitting
outside and wearing sensible clothing, you are well protected indeed.

But take for example the common house mosquitoes in northern America. They
are a culex species, and neem's effectiveness against culex varies. The number
of bites will be greatly reduced, but you probably still get bitten.

So to make an educated guess how effective neem oil will be, it helps to know
what mosquito species are common in your area or in the area you want to visit.
There are many different kinds of mosquitoes and they are specific to different
regions.

Another way to determine how well neem oil works for you as a natural
mosquito repellent is to simply try it out.

And now to one important caveat. The only studies that showed significant
protection from mosquito bites through neem oil, were those that used the neem
oil in coconut oil mix. Rubbing pure oil on your skin is not the most pleasant

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sensation in hot an humid climates where mosquitoes are most active. Oh, and
neem oil does not smell of roses!

Several companies offer personal neem outdoor sprays like this or this They
may not necessarily use neem oil, as neem leaf extract has similar properties
though it is not as powerful as the oil. Those outdoor sprays do offer some
protection against biting insects in general. The neem leaf is also wonderfully
soothing for skin that is irritated from the heat and insects.

But realistically these sprays do not offer quite the protection that you would get
from a neem oil/coconut oil mix. You can not expect them to perform like a
DEET containing mosquito repellent. Also, just like other natural repellents
those neem sprays need to be applied frequently whereas DEET lasts for many
hours.

Natural mosquito repellents do work. I use only natural mosquito repellent, and
I prefer those containing neem because of the many other beneficial effects it
has. For most of the year and around my house the amount of protection those
natural mosquito repellents offer is more than good enough.

But there are times of the year where the mosquitoes are REALLY thick. (I live
in a tropical area.) During those times I make sure I also cover up from head to
toe, avoid the wettest and most overgrown areas in the garden and avoid being
outside at dusk and dawn. I will still not put any DEET on my skin. That stuff in
my opinion is far too toxic to use it regularly.

Don't count on neem natural mosquito repellent to perform a miracle. But use it
right (i.e. using the coconut oil/neem oil mix, or using a spray and applying it
regularly, and also burning neem in a lamp, candle or flare) and combine that
with other common sense measures, like covering up and using mosquito
netting, and there really is no need to put DEET on your skin.[29]

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Neem oil method:-

The method of processing is likely to affect the composition of the oil , since
the methods used , such as pressing (expelling) or solvent extraction are
unlikely to remove exactly the same mix of components in the same proportions
. The Neem oil yield that can be obtained from Neem kernels also varies widely
in literature from 25% to 45%. The oil can be obtained through pressing
(crushing) of the seed kernel both through cold pressing or through a process in
corporating temperature control . Neem seed oil can also be obtained by solvent
extraction of the Neem seed , fruit , oil cake or kernels . A large industry in
India extracts the oil remaining in the seed cake using hexane . this solvent
extracted oil is a lower quality as compared to cold pressed oil and is mostly
used for soap manufacturing . Neem cake is a product obtained in the solvent
extraction process for Neem oil.

Method:- Preparation of neem extract :

The whole neem tree contains bitterness in varied extent but higher
concentration of it found in the neem kernel . Neem kernel is a valuable source
of major limnoid . It is there for essential to understand the scientific method of
fruit collection and depulping to get kernels . 16 (1-8-5) Collection of Neem
fruit :-

The Neem yields fruits during May to August every year . being rich in
carbohydrates neem fruit gets attacked by fungi when come in contact with
soil . such fruits may damage the quality of the final products prepared from
these fruits . hence it is strongly recommended to avoid contact of Neem fruits
with soil . as the fruit ripes they must be depupled as early as possible. (1-8-6)
Depulping of Neem fruits:-

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Depulping is a process to remove seed coat and pulp from the Neem seed . It is
done by hand and using mechanical depulper .Rub the ripe Neem fruits between
palms in the bucket of water and wash the seed . by use clean water for
depulping . Neem Research and Technology Development Center (NRTDC)
developed a mechanical depulper to handle large quantity of Neem fruits . After
depulping and cleaning the Neem seeds in a cool and dry place . if process
properly these neem seeds can be stored for about 6-12 months (1-8-7)

Separation of the kernels:- The dried Neem seeds are grand slightly by the hand
and the outer shell of the seed is removed , kernels are present inside the shell
which are separated and then made into powder using a grinder . it should be
powdered such this no oil comes out of it . This coarse powdered is used for
further studies for the extraction of azadirachtin ."willard1988" (1-8-8)
Extraction of azadirachtin from Neem kernel powder:- For extraction of
azadirachtin by solvent process weight about 500 g of fine tea powder sized
clean neem kernel powder . it should be pounded in 17 such away that no oil
comes out of it . make a thimble and fill it with kernel powder , place it in a
soxhlet apparatus and add about 600 ml of di-chloromethane . keep it in a
heating mantle and heat the reflux for about 12 hours, when the powder kernel
is extracted with solvents like di-chloro-methane or ethyl acetate liminoides and
other constituents get dissolved in it . leaving the seed cake without any active
components . The solvent from this mixture is recovered by distillation . the
distillation or concentrated solution is kept a side for cooling hexane is added to
the concentrate and then filtered using vaccum pump . the above residue is dried
which gives a pale greenish colored powder . this powder consist of
azadirachtin and a very small quantity of nimbin . 100 g of neem kernel powder
on extraction gives about 1 g of azadirachtin ."Patrizia" (1-8-9)

TLC confirmation :-

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 Take a small amount of azadirachtin and dissolve it in methanol , spot the
sample on a TLC plate and place it in a beaker consisting iso-propanol :
hexane(2.5 : 17.5) as mobile phase . Run till 3/4th of the plate , then place it in
iodine chamber . it gives two pale yellow colored spots . the upper spot
indicates azadirachtin and the below spot indicates nimbin . "Hui-Pring Huang"

UV-Spectrophotometer:-

Different concentrations of Azadirachtin were prepared with different solvents

and scanned in the UV region . the wave length 220nm was selected for
Azadirachtin where it show maximum absorbance . lower concentrations does
not show any absorbance hence concentrations (above 10mcg) were taken for
calibration . the calibration curves were linear over the concentration range of
30-70 Mg/ml of Azadirachtin . absorbance were plotted versus respective
concentrations . "Otmer" 18 (1-8-11) High performance liquid chromatography

HPLC :-

The retention time of Azadirachtin is determined using HPLC . The column

used for chromatographic separations was reverse phase phenomena . a hence
the analytical wave length of 210nm was set and the sample was injected . The
chromatographic separation were accomplished using mobile phase consisting
of acetonitryl : methanol : 1% triethyl amine adjusted to PH 4 (60:40:1) ,
filtered through a filter using value stage vaccum pump . mobile phase was
pumped at a flow rate of 1ml/min at room temperature . "[30-39]

Reason for using camphor as mosquito repellent:

Camphor is an easily available ingredient that kills mosquitoes. Made from a

tree extract, this compound has been found to have the

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longest mosquito repellent effect compared to other natural products.
Burning camphor in a closed room can be extremely effective in

Preparation Procedure of Effective & Healthy Mosquito Repellent

Malaria and Dengue are on a steep rise. We all must be using liquid or the old
blue tablet type of mosquito repellents sold under various names. Despite their
use, it is common to see mosquitoes hovering around with the best of repellents
in action. Secondly, we cannot use these mosquito repellents whole day – be
they be of any brand. Thirdly, many people land up with allergies due to them.

In the past, man would light diyas and burn camphor on a regular basis as a part
of daily puja. These helped to purify the air and keep harmful bacteria, viruses
and mosquitoes away! We can definitely have a better and healthy environment
with the use of camphor.

Camphor or Kapur is a waxy, white or transparent substance extracted from the

wood of the Camphor Laurel tree found in Asia. Camphor has many known
medicinal uses-
It relieves nasal congestion and cough when rubbed on the chest as an oil.

Camphor is used as an ingredient in throat lozenges and cough syrups and in

Vicks. Camphor is also used in some anti-itch ointments, creams and cooling
gels because it is can be absorbed through skin and is effective at treating pain
locally. It has an analgesic effect which makes it a favourite oil to be used in
pain relieving massage blends for sore muscles and arthritic pain. Camphor is
known to improve the quality of air making it a better for our lungs and heart.

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Many people are not aware of the Mosquito repellent power of Camphor - a
simple solution.[40]
Preparation Presentation

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Reference:
1. Abiy, E., Gebre-Michael, T., Balkew, M., Medhin, G. (2015). Repellent
efficacy of DEET, MyggA, neem
2. (Azedirachta indica) oil and chinaberry (Melia azedarach) oil against
Anopheles arabiensis, the
3. principal malaria vector in Ethiopia. Malaria Journal, 14(187), 1–6.
4. Adeeko, K. A., Ajibola, O. O. (1990). Processing factors affecting yield and
quality of mechanically
5. expressed groundnut oil. Journal of Agricultural Engineering, 45, 31–43.
6. Ahmad, S., Ansari, M. S., Muslim, M. (2015). Toxic effects of neem based
insecticides on the fitness
7. of Helicoverpa armigera (Hubner). Crop Protection, 68, 72–78.
8. Al-Quraishy, S., Abdel-Ghaffar, F., Mehlhorn, H. (2015). Head louse control
by suffocation due to
9. blocking their oxygen uptake. Parasitology Research, 114(8), 3105–3110.
10. Ambethgar, V. (2015a). Field evaluation of some insecticides against white-
tailed mealy bug, Ferrisia
11. Chatterjee R, Singh O, Pachuau L, Malik SP, Paul M, Bhadra K (2010)
Identification of a sulfonoquinovosyldiacyl glyceride from Azadirachta
indica and studies on its cytotoxic activity and DNA binding properties.
Bioorg Med Chem Lett 20(22): 6699-6702.
12. Devakumar C, Sukhdev S (1996) Neem. In: Randhawa and Parmar B S,
Eds, 2nd Edition, Society of Pesticide Science, New Delhi 77-110.
13. Elavarasu S, Abinaya P, Elanchezhiyan S, Thangakumaran, Vennila K,
Naziya KB (2012) Evaluation of anti-plaque microbial activity of
Azadirachta indica (neem oil) in vitro: A pilot study. J Pharm Bioallied Sci
4: S394-S396.

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14. Galhardi LC, Yamamoto KA, Ray S, Ray B, Linhares RE, Nozawa C
(2012) The in vitro antiviral property of Azadirachta indica polysaccharides
for poliovirus. J Ethnopharmacol 142: 86-90.
15. Iffat A, Siddiqui BS, Faizi S, Siddiqui S (1989) Structurally novel
diterpenoid constituents from the stem bark of Azadirachta
indica(meliaceae). J Chem Soc Perkin Trans 2: 343–345.
16. Adewoye, T. L., Ogunleye, O. O. (2012). Optimization of neem seed oil
extraction process using
17. Response surface methodology. Journal of Natural Sciences Research, 2(6),
66–76.
18.https://www.google.com/search?
rlz=1C1OKWM_enIN800IN800&biw=1455&bih=717&ei=FmScXPCDN9n
orQGGo6z4BQ&q=various+marketed+avilable+mosquito+repellent&oq=va
rious+marketed+avilable+mosquito+repellent&gs_l=psy-
ab.3...11083.15301..16314...0.0..0.177.1244.0j8......0....1..gws-
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2001.

21.C. Sinha, A.K. Agrawal, F. Islam, K. Seth, R.K. Chaturvedi, S. Shukla and P.K.
Seth, Mosquito repellent
22. (Pyrethroid based) induced dysfunction of blood-brain barrier
permeability in developing brain,
23. International Journal of Developmental Neuroscience, 22 (1), 31-37
(2004).
24.A.A. Odetola and O. Bassir, Evaluation of anti-malarial properties of some
Nigerian medical plants.
25. Nasci, R. S., Haria, C. W. and Porter, C. K., Sangyo Eiseigaku Zasshi, 1995,
37, 5–8. 2. Belton, P., Mosq. News, 1981, 41, 751.
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26. Curtis, C. F. and White, G. B., New Sci., 1982, 93, 328.
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1992, 429–436.
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organic-repellents
29. 1.a-b-c- Anna Horsb rugh poter ,17April 2006, Neem india’s.
30. 2.a-b-c-d- USDA Grin taxonomy.
31. 3.a-b-c- Neem,Web MD.
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through herbal drugs in rural India , Fitoterapia.Vol –LXL No 3.240 -242.
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dirachtin india leaf “Melia azadirachta” Journal of the india chemistry
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repellant_561320_blog

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