2. REVIEW OF LITERATURE 2.1 Introductionshodhganga.inflibnet.ac.in/bitstream/10603/43421/9/09...Review of Literature Figure-2.3: Ajanta caves Figure-2.4: Mural Painting of Ajanta caves

A Study on Microbial Deterioration of Ancient South Indian Mural Paintings and

Fabrication of Biobased Natural Protective Coating for its Conservation --------------------------------------------------------------------------------------------------------------------------------------------------------------------

6 Review of Literature

2 REVIEW OF LITERATURE

21 Introduction

We are very well aware that there are thousands of wall paintings existing in

various part of India dating from the prehistoric times to the present date Wall paintings of

Ajanta Ellora and Bagh are world famous Besides these well known examples there are

thousands of other mural paintings which are extremely important but are less

known (Agrawal and Pathak 2001)

Murals are paintings made on walls of caves and palaces while miniatures are

small-sized colorful intricate handmade illumination Murals are considered to be the

earliest evidence of Indian paintings which are unearthed as the symbol of ancient

civilization The term mural is derived from the Latin word ldquomurusrdquo which means wall

The paintings carried out on the walls are known as mural paintings (Mini 2010) Mural

paintings reflect the feelings of Indian spirituality prismed through the age-old mythologies

and spirituality

22 History of wall and mural painting

221 History of wall painting

The history of art of painting in India can be traced back to pre historic and proto

historic times The earliest specimen was found in caves and rock shelters found admits

rocky hills These paintings are dated from 1200 BC to the tenth century AD (Mathpal

1984 Misra et al 1977) Artifacts found at Mohenjodaro and Harappa has traces of

natural pigments suggestive of the use of colors in pre-Aryan art These cave paintings has

quite a wide distribution though all the paintings do not belong to one period Rock shelter

paintings are usually seen on back walls arranged in horizontal lines and often with many

super impositions of successive styles The earliest paintings go back to at least 550 BC

Ajantan art the tradition of which extends more than two millennia from the second

century BC to the present represents the flowering of Indian mural paintings History of

wall paintings in India is studied through its evolution from Ajanta to Kerala Ajanta

paintings considered to have been executed between the 2nd

century BC and 5th

centaury

AD Murals of 6th

centaury AD was found at Badami (Karnataka) and Sittannavasal Jain

cave temple (Tamilnadu)Murals of Narthamala Vijayalaya Chozeswara belongs to

9th

century AD and murals of Brihadeeswara temple Tanjore (Tamilnadu) belongs to




11th

century AD Murals seen in Kerala are dated between 16th

17th

and 19th

century AD

(Sasibhooshan 2006)

The most important sites of rock paintings in North India are at Bimbetka Gwalior

Tikla Ayra Bhanpura Rampura Singhauli Bhojpur Adamgrh Delware and Mirzapur

In south India they are found at Mehaboob Nagar (Andhrapradesh) Edakkal Marayoor

and Perimkadavila (Kerala) Mallampadi (Tamilnadu) and Bellary (Karnataka)These

paintings were considered as the early models of mural paintings The mural painting of

Sittannavasal Badami Lepakshi Tanjore Vijayanagar and Kerala has a lot of similarities

Style colour theme and technique of mural painting varied with different dynasties their

ruling period and important painting sites are given belowThe important mural painting

site Ajantha cave 9 amp10 was made during the ruling period from 2nd

BC to 2nd

AD of

Satavahana dynasty Vaishnava cave in Badami in the ruling period from 6th

-8th

AD

belongs to early western Chalukyas During 7th

-9th

AD Pallava dynasty in Kailasanatha

temple Kancheepuram was considered to be as one of the dominant mural painting site in

Southern India Similarly other South Indian dynasties early Pandya between 7th

and

9th

AD and early Chera from 8th

to 9th

AD highlighted their mural paintings at

Sittannavasal cave temple and Thirunandikkara cave temple respectively Mural paintitngs

at Kailasanatha temple Ellora from 8th

- 10th

AD was made by the dynasties of

Rashtrakuta The familiar mural paintings at Brihadeeswara temple Thanjavur and

Vijayalaya Choliswaram temple Narthamalai was carried out by the chola dynasty

between the period of 9th

and 13th

AD Another biggest dynasty called Vijayanagara

betweeen 14th

and 17th

AD made numerous mural paintings at Vidharbha temple

Lepakshi Varadaraja Perumal temple Kancheepuram Vitthala temple Hampi and Sri

Ranganatha tenple Srirangam Nayakarsquos dynasty ruled inthe period from 17th to 18th AD

established the mural paintings in Kapardisvara temple Tiruvanjuli and later murals of

Thanjavur and Kancheepuram Finally medivial Kerala (blend of Hoysala Chalukya and

Vijayanagara paintings) in 16th

and 18th

AD carried out the mural paintings at Tripayar

temple Ettumanur temple Thiruvanchikulam temple Panayanarkavu temple

Mattancherry Padmanabhapuram and Krishnapuram palace (Sivaramamurthi 1994)




222 History of mural painting

The first paintings were Cave Paintings Ancient peoples would decorate walls of

protected Caves with paint made from dirt or charcoal mixed with spit or animal fat In

Cave paintings the pigments (often carbon black or ochre) stick to the wall partially

because the pigment gets trapped in the porous wall and partially because the binding

media (the spit or fat) dries adhering the pigment to the wall Over the years countless

graves unearthed by archaeologists exposed bodies covered in red pigment or chunks of

pigment buried alongside the bodies Red associated with blood the most life sustaining

of body fluids was the appropriate color to symbolize lifes meaning and end The word

Hematite (the source of many iron oxide pigments) is derived from the Greek word hema

meaning blood (Chakarabarti 1980)

Wall paintings have been cultural expressions of human creation throughout

history from the earliest beginnings such as rock art extending up to present day murals

Throughout history humans have used the most beautiful and durable stones for

monumental buildings Beginning in the sixth century BC marbles extracted from the

Pentelikon quarry became a common building material Abundant limestone and marble

quarries enabled the ancient Greek architects to build the Acropolis and Roman architects

to build the Forum (Saiz 2002)

Figure-21 Cave painting




Figure-22 Mural painting

The oldest known Maya murals are also the most recently discovered Murals

found at the Preclassic site of San Bartolo Guatemala date to 100 BC The church of San

Pietro and the ldquooratoriordquo of San Benedetto are believed by most scholars to date from the

third quarter of the 11th century During the last decade of the same century that some

changes were made to the edifice of the church and it was decorated with mural paintings

and stucco reliefs deep in the jungle of southeastern Mexico archaeologists were surprised

that the murals are about 1350 years old

St Agatharsquos Complex was one of the preferred sites since it consists of the

hypogean environment embellished by a significant number of wall paintings some dating

back to the fourth century AD The Catacombs of St Paulrsquos constitute a complex of

interconnected underground Roman cemeteries that were in use up to the 4th century AD

There are a few surviving murals and ochre inscriptions from late Roman and early

medieval periods Decorated tablets and reliefs adorn the graves (Zammit et al 2008) The

uncovering of paintings was made from the middle of the 19th century until the 1960s

23 Indian murals

History of wall paintings in India is studied through its evolution from Ajanta to

Kerala Ajanta paintings considered to have been executed between the 2nd

century BC and

5th

century AD Murals of 6th

century AD was found at Badami (Karnataka) and

Sittannavasal Jain cave temple (Tamilnadu) Murals of Narthamala Vijayalaya Chozeswara

belong to 9th

century AD and murals of Brihadeeswara temple Tanjore (Tamil Nadu)

belong to 11th


17th and

19th

century AD (Sivaramamurthy 1994)




India has one of the greatest traditions of painting of the ancient world Murals and

miniature are the two different forms of arts which intrinsically related to Indian painting

tradition The earliest evidence of murals is the beautiful frescoes painted on the caves of

Ajanta and Ellora also on the Bagh caves and Sittanvasal The oldest mural paintings date

is about 3000 years BC (Sasibhooshan 2006)

The number of wall paintings in each state of India is very large and they are

suffering from all types of deterioration and damage but in comparison facilities for their

preservation are much too small This is a very serious problem The main objective of the

present study is to develop a protective bio-based gel (Rajagopalan 2008)

24 North Indian murals

North India has a rich heritage of Mural paintings even before the Mughal period

The murals at the Vishnu Temple located at Madanpur in Lalitpur district of Uttar Pradesh

of 12th century AD reveals the skillful hands of the painters In Eastern India there are

many evidences of wall and panel paintings describing Buddhist and non- Buddhist

themes In Arunachal Pradesh and Tripura sublime mural works have been found which

needs further study Ladakh is known for its wall paintings in Alchi and Hemis

monasteries made on 11th-12th century and the Spiti Vally in Himachal Pradesh is known

for its Buddhist paintings in the Gomphas of Tabo Monastery (Havell 1982)

Ajanta is known to be the fountain head and inspiration of Buddhist paintings

across the whole of Asia There are fragments of paintings of the time of Ajanta which

survive at many Buddhist cave sites including Pitalkhora near Ellora in Maharashtra The

magnificent Kailashnath temple had been hewn out of a mountain at Ellora in the 8th

centuryThe Kashmiri style was mainly responsible for the lovely wall paintings still seen

in the beautiful monasteries at Alchi Mangyu and Sumda in Ladakh in the Tabo

monastery in the Spiti valley and in the Nako monastery in Kinnaur district Himachal

Pradesh On the western edge of the trans-Himalayan plateau in Spiti is the monastery

complex of Tabo This appears to be one of the first among the 108 monasteries built by

Yeshe It is dated around 996 AD There are also paintings of the late 9th century in the

Jain caves at Ellora (Spink 2009)




Figure-23 Ajanta caves

Figure-24 Mural Painting of Ajanta caves

Ellora Caves Maharashtra belong to the highest achievements of mankind both

regarding the engineering skills and arts Some 2000 years ago Pitalkhora Caves

(Maharashtra) were among the largest temple complexes in India These 11 - 14 caves

were created in 250 BC - 4th century AD In 4th century AD temples were adorned with

beautiful murals especially fine paintings have been preserved on columns of Cave 3

Traces of murals are in Cave 6 as well Already the oldest group of caves here - Buddhist

caves from 630 - 700 AD - contain paintings of Buddhist panteon Remnants of paintings




are seen in the most beautiful temple - Cave 16 Kailasanatha circa 760 - 860 AD Some

Jain temples - Cave 32 (Indra Sabha) Cave 33 - contain remnants of valuable murals from

800 - 1000 AD - this artwork marks the end of ancient period in the Indian art and

beginning of medieval art (Baskaran 1975)

Figure-25 Ellora caves

There are surviving mural paintings from the 17th century onwards in Rajasthan

The verdant Pahari hills saw the finest continuation of the tradition of murals in India The

18th and 19th century paintings on the walls of the Rang Mahal in Chamba are among the

best surviving examples of Pahari murals The 18th century paintings on the walls of the

Viranchinarayan Temple at Buguda are some of the finest surviving murals of that period

in India

Figure-26 Paintings on the walls of the Rang Mahal in Chamba






In South India they are found at Mehaboob Nagar (Andhrapradesh) Edakkal Marayoor

Perimkadavila (Kerala) Mallampadi (Tamilnadu) and Bellary (Karnataka)These

paintings were considered as the early models of mural paintingsThe mural painting of


(Sasibhoosan 1987)

25 South Indian murals

South India also got rich tradition of mural paintings In the reign of Cholas

Vijayanagaras and Nayakas this art reached the climax The Deccan art of Bijapur

Hyderabad and Golconda schools were influenced by the Mughal traditions and later by

European idiom Maratha murals are also shaped under the Mughal traditions and

employed oil as medium The Mural art of Kerala vividly depicted on the walls of temples

and monuments show the traces of European affinity In Tamil Nadu almost all the extent

examples of frescos have been done on rock surfaces and stone walls This is a crucial

factor in determining the quality of frescos (Havell 1982) The ceiling of the Virupaksha

temple in Hampi is covered with paintings of the 15th century The paintings on the ceiling

of the mandapa of Lepakshi temple are some of the finest mural paintings of the medieval

period in India Legends associated with Siva and Parvati Krishna and Rama were painted

on the walls of palaces and temples in Kerala from the 16th to 19th century Mural

paintings were made on the walls of all the palaces within the magnificent Orccha fort The

Raj Mahal was completely adorned with mural paintings of the 17th century

Figure-27 Virupaksha temple in Hampi




Figure-28 Ceilings of Virupaksha temple Hampi

The murals on the walls of the navaranga (the central pillared hall) depict stories

from Bhagavatha and are excellent examples of 19th

century Mysore paintings The

Mallikarjunaswamy temple on the hill is one of the Panchalingas of Talakad and the

temple has a beautiful mural painting on the wall of the temple The Dariya Daulat Bagh

has excellent mural painting (Sasibhoosan 1987)

Figure-29 Paintings on the walls of Jagan mohan palace Mysore




Figure-210 Painting of Raja woodeyar era

Kerala mural paintings are the frescos depicting mythology and legends which are

drawn on the walls of temples and churches in South India principally in Kerala Ancient

temples churches and palaces in Kerala South India display an abounding tradition of

mural paintings mostly dating back between the 9th to 12th centuries The murals of

Thirunadhikkara Cave Temple (now ceded to Tamilnadu) and Tiruvanchikulam are

considered the oldest relics of Keralarsquos own style of murals Fine mural paintings are

depicted in temples at Trikodithanam Ettumanur Vaikom Pundarikapuram

Udayanapuram Triprangode Guruvayoor Kumaranalloor Aymanam the Vadakkunathan

temple in Trichur and the Sri Padmanabhaswamy temple at Thiruvananthapuram Other

mural sites are in the churches at Edappally Vechur Cheppad and Mulanthuruthy and at

palaces such as the Krishnapuram Palace near Kayamkulam and the Padmanabhapuram

Palace

Figure-211 Kerala murals




Figure-212 Murals paintings of Kerala temples

There are wall paintings in Badami in the Bijapur district Karnataka Some

excellent paintings exist in the Jain caves at Sitanavassal in Puddukkottai District of Tamil

Nadu Tamil Nadu gave exuberant and glorious expression to themes relating to Siva in the

temples of Panamalai and Kailashanatar in Kancheepuram In the 9th century Jain cave of

Sittannavasal in Tamil Nadu there is a marvellous lotus pond painted on the ceiling In

the heart of the Brihadeeswara temple in Thanjavur Tamil Nadu protected by massive

walls of stone are the finest paintings of the theme of Siva ever painted towards the end of

the 10th century (Sivaramamurthi 1992)

Figure-213 Kancheepuram Mural paintings




Figure-214 Sculpture of Mahabalipuram

26 Methods of mural painting

In the history of mural several methods have been used The Buon fresco technique

consists of painting in pigment mixed with water on a thin layer of wet fresh lime mortar

or plaster The pigment is then absorbed by the wet plaster after a number of hours the

plaster dries and reacts with the air it is this chemical reaction which fixes the pigment

particles in the plaster After this the painting stays for a long time up to centuries in fresh

and brilliant colors A Secco painting is done on dry plaster The pigments thus require a

binding medium such as egg (tempera) glue or oil to attach the pigment to the wall

Mezzo-fresco is painted on nearly-dry plaster which is defined by the sixteenth-century

author Ignazio Pozzo as ldquofirm enough not to take a thumb-printrdquo so that the pigment only

penetrates slightly into the plaster Ancestral Hopi village archaeologists found that

fourteen kivas had painted murals and multiple layers of murals on the walls some with up

to twenty-seven layers of murals (Saturno 2006)

The colors used in painting are generally obtained from plants and minerals

Among them only such colors which are lime resistant are suitable for mural painting The

selection of proper colors for a mural painting is an important part of the work as the

longevity the painting depends on it Only mineral colors are preserved for a long time

which could be the reason for use of those colors in early days As per ldquoShilpa ratnardquo and

ldquoAgnipuranardquo the colors used in mural painting are white black red yellow blue and

green Yellow and red are from yellow ochre and red ochre Blue is generally indigo or

china blue White is the prepared wall and black is the carbon soot Mural paintings have




survived centuries only on account of the identification and use of those colors which does

not have any reaction with lime (Miraseth 2006)

Originally three types of brushes were made and used flat medium and fine Flat

brushes were made from hairs behind calfrsquos ear the medium from the bottom of goats

belly and the fine brush from delicate blades of grass However in due course of time and

change use of easily available brushes from commercial market and shading of the

original colours can be distinctively noticed The use of blue colour is also noticed over the

passage of time The paintings have been brought down from the walls to plywood cloth

paper and other frames for easy to make and use purposes (Chakarabarti 1980)

27 Themes of mural painting

The subjects and themes in the murals are centered on the Hindu pantheon of Gods

and Goddesses drawn from description in the invocatory verses or Dhyana Slokas More

common representations are of Vishnu in different incarnations Ganesha and

manifestations of Siva We see murals in the Pallimanna Siva Temple devoted to Siva-

Parvatirsquos marriage the Ramayana Story being narrated on the walls of the Mattancheri

palace in the Sankara Narayana the Mahabharata is retold Murals are various scenes from

the Hindu epics and also the frolics of Lord Krishna

It would be interesting to bring into account the influence of the mural paintings on

Christianity On various churches like the St Georgersquos Orthodox Church has Christian

murals bringing out biblical characters and narrating stories from the scriptures Mar

Sabore and Afroth Church at Akapparambu in the outskirts of Ernakulam also have murals

of scenes from the bible Interestingly StMaryrsquos Church at Kanjoor has on both sides of its

entrance a battle scene between the armies of Tipu Sultan and of the English East India

Company It is also apparent that the history of Kerala Murals has its origin in the temple

walls and thus a study of Kerala Murals is not complete without pondering on the

architecture of Kerala its association with the murals (Warrier 2011)

28 Technique used in mural paintings

Indian murals executed on walls of caves structural temples walls of palaces were

based on highly developed techniques Mural painting is a composite structure of these

three basic layers the support the ground and the pigment layer In case of wall painting

the wall itself is the support This is followed by the layer or layers of plaster which




constitute the ground The pigment layer is the one which contains the picture The support

is the wall which may be rock surface in a cave or a structural one These stone surfaces

which serve as carriers are rough or uneven This unevenness is itself an advantage as it

provides ldquotoothrdquo to hold the plaster on the surface The uneven surface of the wall has to be

made uniform with layers of plaster in successive gradation of fineness There is generally

one layer of rough plaster followed by one or more layers of fine plaster Ground or plaster

is a term used to mean the surface material of a wall or building which is applied as a

plastic mass made by mixing certain materials with water letting the mass set by drying

carbonation or hydration The active or setting components of plaster are commonly clay

which sets by drying lime which sets by carbonation gypsum and pozzolana which sets by

hydration (Mini 2010)

There are different mixtures for rough plaster In all the mixtures clay limestone

lime and shells in powdered form serve as the matrix The particles of which are held

together by the organic materials like gum glue extracts from barks etc are in fact

adhesives The fine plaster layer requires more careful preparation because it is the layer on

which the pigments are laid to make the picture ldquoShilparatnardquo suggest a mixture of conch

Oyster shells or white clay with gum of neem or slaked lime and coconut water Here the

consolidation is effected by the carbonation of slaked lime ie it absorbs carbon dioxide

from air and becomes carbonate Mural paintings were lime-washed from the 17th until the

beginning of 19th century A few paintings in vaults were never lime-washed for example

at Taby and Floda painted by the medieval painter Albertus Pictor (Mini 2010)

All the paintings have certain basic common characteristics There has to be a wall

or rock surface called support over which the artist goes to work A plaster base called

ground of varied thickness is laid on the support The ground may consist of one or more

layers The strength and life of the whole mural depended up on this ground over which the

painting was done It is the process of applying the final painting on the ground determines

the character of the mural whether it is fresco or tempera The mural painting is done

usually by creating wall surface free of any previous oil or paint by using solvent or by

sand blasting The brick wall is first plastered with mixture of lime and sand in the ratio of

12 A mixture of egg yolk vinegar oil colour is used in dry plaster

There are at least six stages before a Kerala Mural Painting is complete Lekhya

Karma is the first stage where sketching by pencil or crayon is done The second stage is




the Rekha Karma which enhances the outlines of the sketching done The third stage called

Varna Karma breathes life into the painting with its colours Here all colours are

appropriately filled in The colouring of the human figures is done according to the

characters and their common virtues The divine and noble characters (Saatvik) are painted

in green those inclined towards power and wealth (Raajasik) is painted in shades of red

lowly mean characters (Tamasik) in white and demons in black In the fourth stage or the

Vartana Karma shading is done Fifth stage called the Lekha Karma brings us to the final

outline of the painting which is usually in black The final and the finishing touches stage

are called Devika Karma (Warrier 2011)

29 Dyes used in mural painting

Indians have been considered as fore runners in the art of natural dyeing Natural

dyes find use in the colouring of textiles drugs cosmetics etc Owing to their non-toxic

effects they are also used for colouring various food products In India there are more

than 450 plants that can yield dyes In addition to their dye-yielding characteristics some

of these plants also possess medicinal value Though there is a large plant resource base

little has been exploited so far Due to lack of availability of precise technical knowledge

on the extracting and dyeing technique it has not commercially succeeded like the

synthetic dyes (Siva 2007)

The color materials on the mural paintings in ancient India were derived from the

natural materials like terracotta chalk red ochre and yellow ochre mixed with animal fat

The dyes for painting is prepared such as white from lime Black from carbon soot of

lamps Red and yellow from minerals Blue from indigo ferra Deep red from lac A fine

coating of resin is done after the painting in order to make a shining appearance Egg yolk

and vinegar was mixed with oil colors and painted on dry plaster (Krishnakumar 1999)

Although plants exhibit a wide range of colours not all of these pigments can be

used as dyes Some do not dissolve in water some cannot be adsorbed onto fibres whereas

others fade when washed or exposed to air or sunlight It remains a mystery why plants

reward us with vibrant dyes India has a rich biodiversity and it is not only one of the

worldrsquos twelve mega diversity countries but also one of the eight major centres of origin

and diversification of domesticated taxa It has approximately 490000 plant species of

which about 17500 are angiosperms more than 400 are domesticated crop species and




almost an equal number their wild relatives Thus India harbours a wealth of useful

germplasm resources and there is no doubt that the plant kingdom is a treasure-house of

diverse natural products One such product from nature is the dye (Siva 2007)

Many natural dyestuff and stains were obtained mainly from plants and dominated

as sources of natural dyes producing different colours like red yellow blue black brown

and a combination of these Almost all parts of the plants like root bark leaf fruit wood

seed flower etc produce dyes It is interesting to note that over 2000 pigments are

synthesized by various parts of plants of which only about 150 have been commercially

exploited Nearly 450 taxa are known to yield dyes in India alone of which 50 are

considered to be the most important ten of these are from roots four from barks five from

leaves seven from flowers seven from fruits three from seeds eight from wood and three

from gums and resins Some important dye-yielding plant habitats their distribution and

colouring pigment (Siva 2007)

The dye is generally prepared by boiling the crushed powder with water but

sometimes it is left to steep in cold water The solution then obtained is used generally to

dye coarse cotton fabrics Alum is generally used as a mordant Flowers of Butea

monosperma (Lam) Taubert yield an orange-coloured dye which is not fast and is easily

washed away For the purpose of colouring the material is steeped in a hot or cold

decoction of the flowers A more permanent colour is produced either by first preparing the

cloth with alum and wood ash or by adding these substances to the dye-bath The dye

indigo is produced by steeping the plant in water and allowing it to ferment This is

followed by oxidation of the solution with air in a separate vessel Mallotus philippinensis

muell yields an orange colour used for dyeing silk and wool To prepare the annatto dye

from B orellena the fruits are collected when nearly ripe The seeds and pulp are removed

from the mature fruit and macerated with water Thereafter they are either ground up into

an lsquoannatto pastersquo or dried and marketed as annatto seeds Sometimes when the seeds and

pulp are macerated with water the product is stained through a sieve and the colouring

matter which settles out is collected and partially evaporated by heat and finally dried in

the sun (Sowbhagya 2002)

The predominant colors used in rock art and cave art are black (from charcoal soot

or manganese oxide) yellow ochre (limonite) red ochre (hematite or baked limonite) and

white (kaolin clay burnt shells powdered gypsum or powdered calcium carbonate) All of




theses pigments may be used on leather or raw hide as well Early cave art in Seminole

Canyon was monochromatic in red and as culture grew more sophisticated the color

palette expanded Blue Orange Brown Green and Purple are the most common colours

used in Ajantha murals White Pink Gold are occasionally represented mineral earths

containing ferric oxide and copper carbonates were used as source of red yellow and green

colours There is also evidence to establish that root of Rubia cardifolia and leaves of

Indigofera tinctoria were used as source for red and blue colours (Nayer et al 1999)

Many colors of pigments were obtained from both organic and inorganic sources in

ancient times and used to paint various surfaces such as braintan leather clothing and

accoutrements rawhide the human body etc Blue color was obtained from powdered

azurite and lapis duck manure and certain clays Fine Blue-green clay was obtained from

deposits near Mankato Michigan Organic sources of blue include Turkeys Tail a

greenish blue polypore that grows on dead decaying wood (not tannish ones) Native

Americans get Indigo from either Guatemala or Ecuador which was traded well up onto

Kansas long before the Spanish ever set foot here American Indigo has purple tones to it

but IndianJapanese indigo has green tones Berries and crushed flowers were also used to

obtain blue color by rubbing the plant material directly on the leather

An organic source of red was Cochineal It was native west of the Mississippi and

is produced from the bodies of small insects that grow on the prickly pear cactus Ladys

Bedstraw gives a good vermillion red color Harvest the roots dry them out clean the mud

etc off of them and then grind the roots really fine then soak them to extract the red

coloring The key here is drying the root first Blood was also used as red pigment Pucoon

roots also yield a red pigment (Smith 2002) Red ochre the reverse original red colours are

due to variations in iron oxide (or) calcite content and to redox conditions in the firing

produced Brown coloured ceramics owe their colour to maghemite and must be

considered as a fabrication defect due to the position of the ceramic piece in a zone where a

reducing atmosphere prevailed in the course of firingRed pigments were produced from

iron oxides like hematite or limonite and from ochre Hematite was the most frequently

used mineral It occurs in the form of aggregates of small red crystals or in the form of

bigger black crystals which are difficult to crush white was obtained mostly from

kaolinite or illite (Clottes et al 1990)




210 Biodetoriation of mural paintings

Historic items often contain natural biodegradable materials and have histories of

exposure to moist conditions These two conditions make them amenable to microbial

attack Microbial colonization may damage the building or the cultural asset

(Salonen 2001) Biodeteriogens are organisms involved in deterioration of artifacts They

are very specific for each type of artifact in accordance with its chemical structure and

environment They also have different nutritional requirements and act directly or

indirectly on the substrate The adhesion of microorganisms to a substrate is the result of

cell hydrophobicity and of excreted polymeric substances contained by sheaths capsules

and slimes (Colao 2005)

The surviving wall paintings and decorations in subterranean environments around

Malta are rather scarce (Buhagiar 1986 1993 1998) These include those at St Paulrsquos

Catacombs (Zammit 2008) St Agatharsquos Crypt and Catacombs (Camilleri 2001)

San Catald Catacombs Tac-Cghaqi Abbatija tad-Dejr (Buhagiar 1993) all in Rabat the

cave-sanctuary in Mellieħa (Buhagiar 1987) and the Ħal Saflieni Hypogeum in Paola

(Pace 2000) Previous studies conducted on wall paintings and decorations in Maltese

Palaeo-Christian Catacombs have involved their manufacturing techniques (Muscat 2005)

However to date limited systematic studies have been conducted into the biodeterioration

in Maltese Palaeo- Christian Catacombs The only such studies ever conducted in Malta

concerned the biodeterioration of works of art in catacombs have been carried out at St

Agatharsquos Crypt and Catacombs in Rabat (Zammit et al 2008 Psaila 2006)

Microorganisms can be responsible for the destruction of monuments buildings

and cultural heritage assets including the decay of concrete cement stones mural

paintings etc These microorganisms are not only related with biodeterioration but water-

damaged indoor building materials are frequently colonized by complex bacterial

communities and may emit mixed bioaerosols into the indoor air (Jimenez 2000) It is

well known that mural paintings are colonized by micro-organisms and can be severely

damaged by such growths (Ciferri 1999) A large number of fungal (gt50) and bacterial

taxa (70) have been detected in indoor environments Over 50 bacterial species and 14

genera novel to science were reported in the past 10 years from indoor air and historic

items indicating bacterigravea in these enviroments are as yet only fragmentarily known

(Salonen 2001)




Traditionally the study of microflora which causes this biodeterioration was based

on classical cultivation methods that were mainly useful to study the physiological

potential of microorganisms but do not provide information on all microbial communities

to be found in excavated artifacts Indeed many studies using culture-independent

techniques which cultivate microorganisms from different environments may often

represent only minor components of the microbial community as a whole It is generally

accepted that cultivation methods recover less than 1 of the total microorganisms present

in environmental samples (Ward et al 1990 Amann et al 1995) Hence microbial

investigations based only on cultivation strategies cannot be regarded as reliable in terms

of reflecting the microbial diversity present in objects of art (Osburn et al 2011)

Early workers suggested that the presence of living microorganisms on stone

increased its susceptibility to damage through their water-binding capacity It has recently

been reported that the mineralogy porosity surface roughness and capacity to collect

water and organic materials control its bioreceptivity and tendency to biodeterioration

(Diakumaku 1995) In addition environmental factors as temperature light pH and

relative humidity affect the number and type of colonising species (Tiano et al 1995) and

hence the progress of colonisation Biofilms on decayed stones consist on mixed microbial

populations embedded in a polymeric matrix Fungi algae filamentous and unicellular

bacteria are found in these complex communities A variety of bacteria have been isolated

from different stone samples confirming the dominance of Gram-positive organisms

especially spore-forming bacteria such as Bacillus and filamentous actinomycetes

(Ripka 2005)

In cultivation studies of deteriorated mural and rock paintings bacterial isolates

have frequently been attributed to the genus Bacillus (Weirich 1989 Tate and Rebrikova

1991 Gonzalez et al 1999 Heyrman et al 1999 Jimenez et al 2000) Other studies that

have included DNA-based techniques to characterize the bacterial community involved in

biodeterioration have shown that a substantial part of the bacterial population is made up of

so far unknown species (Altenburger et al 1996 Rolleke et al 1996 Gurtner et al 2000

Heyrman and Swings 2001) (Heymann et al 2003)

Many potential mycotoxin producers have been detected in air and items from

indoor environments with histories of exposure to moisture for example species of

Acremonium Alternaria Aspergillus Chateomium Fusarium Eurotium Paecilomyces




Penicillium Scopulariopsis Stachybotrys Trichoderma Verticillium and Wallemia

(Peltola et al 2001 Pieckova et al 2001) Not all strains of one species produce

mycotoxins and different strains may produce different toxins Whereas toxin producing

strains are frequent in crops and outdoor vegetation toxin producing isolates from indoor

environments and non living assets have only been reported of the species Aspergillus

versicolor (Nielsen et al 1998) Stachybotrys chartarum Nikulin (1997) and

Trichoderma harzianum (Peltola et al 2001) Streptomyces griseus strains producing

valinomycin occur in environments where humans can get exposed

(Andersson et al 2002) The biological properties and toxicity mechanisms of cereulide

were identical to those by valinomycin (Mikkola et al 1999 Salonen et al 2001)

The genus Paenibacillus was first proposed by Ash et al (1993) and its description

was emended by Shida et al (1997) The genus Paenibacillus is under intensive taxonomic

investigation with ten species having been formally described in the last year Heyrman amp

Swings (2001) isolated a strain originally classified as Paenibacillus sp LMG 19508

from biodeteriorated mural paintings in the Servilia tomb at the Roman necropolis of

Carmona in Seville Spain (Smerda et al 2006) Building stone and wall paintings contain

a variety of hydroscopic salts including carbonates chlorides nitrates sulfates etc

dispersed within the porous material or locally concentrated These salts are solubilised

and migrate with the water in and out of the stone Due to changes in physical parameters

the so called salt efflorescences appear on the surfaces providing extremely saline

environments and building niches for settlement of halophilic microorganisms Halite and

other hygroscopic salts can retain considerable quantities of moisture when entrapped in

the building materials and thus favouring the bacterial growth (Ripka 2005)

It is demonstrated that water-damaged indoor building materials contained

substances of microbial origin extremely toxic to eukaryotic cells and large quantities of

several microbial species known to be potentially toxic or immunopotentiating These

include peptidoglycans Gram-negative endotoxin β-13-D glucans lipoglycans and

teichoic acids and mycotoxins (Jimenez 2000) In turn these microorganisms can mediate

the crystallization process These crystallizations can produce destructive effects Some

salts may crystallize or recrystallize to different hydrates which occupy a larger space and

exert additional pressure producing cracking powdering and flaking

(Jimenez and Laiz 2000)




211 Types of deterioration on mural paintings

The frescos represent a variety of ecological niches where primary colonization is

due to photoautotrophic and chemolithoautotrophic microorganisms followed by

secondary colonization due to heterotrophic bacteria that obtain nutrients from various

sources (metabolites of autotrophic bacteria airborne organic contamination animal faeces

and organic compounds in the paint layers themselves) Microbial-induced deterioration

processes cause structural as well as aesthetic damage to wall paintings such as the

discoloration of materials the formation of crusts on surfaces and the loss of material

(Sarro et al 2006 Osburn et al 2011) Halophilic bacteria and archaea were thought to

be found only in extreme environments like salt lakes or black smokers However it has

been shown that mural paintings and stones are a common habitat for extremely salt

tolerant and moderate halophilic (Ripka 2005) Microbial induced deterioration processes

can cause different kinds of alterations such as discolouration of materials the formation of

crusts on surfaces and the loss of material which leads to structural damage (Ripka 2005)

First of all microorganisms can cause severe problems due to the excretion of aggressive

metabolic products such as organic or inorganic acids Secondly material components can

be used as substrates for microbial metabolism Exoenzymes such as cellulases proteases

tannases beta-glucosidase produced by fungi and bacteria are responsible for hydrolysis of

cellulose and proteins Further examples are mechanical processes caused by swelling and

shrinking of slime or mycelia penetrating inside plaster of the painting resulting in loss of

cohesion and detachment of the paint layer (Rolleke 1996 Gurtner et al 2000)

Table-21 Microbial damage on mural paintings (Rolleke 1996)

S No Damage Microbial activity

1 Green brown red grey

surface layers (patinas)

Pigments produced by microorganisms (intra-or

extracellular) as chlorophyll and carotinoids

2 Dislodging of painting layers Use of organic binders as nutrient and energy

source

3 Acid attack of carbonates Acid produced by fungi or bacteria

4 Discolouration of pigments Redox-reactions induced by fungi and

bacteria

5 Loss of paint layer Growth of microorganisms deep into the plaster

6 Hydrolysis of synthetic

polymers Chemoorganotrophic activity of bacteria and fungi




Discolorations are caused by excretion of pigments and microbiologically induced

reductionoxidation-processes (Krumbein et al 1990) Degradation of the bindings is

followed by reduction of stability Oxidation of ammonium and reduced sulphur

compounds results in acidification of the environment such as Sulphuric acid and the

variety of organic acids produced by the heterotrophs cause solubilization of calcareous

binding material and chelation of metal ions (Bock et al 1993) It is now well recognised

that wall paintings can be deteriorated by microorganisms (Ciferri 1999)The growth of

microorganisms essentially results in two types of damage aesthetic and structural

Aesthetic damage implies pigment discoloration stains and formation of a biofilm on the

painted surface Structural damage is observed as cracking and disintegration of the paint

layer formation of paint blisters and degradation of support polymers or of glues and

binders resulting in detachment of paint layer from the support The colonization of mural

paintings by bacteria and other microorganisms can have different causes Provided that

favorable environmental conditions occur nutrients for heterotrophic bacteria and fungi

can be available from metabolites of autotrophic bacteria from airborne contamination and

dripping water from animal faeces but also from organic compounds in the paint layer

itself

212 Methods of studying biodeterioration

It is now well recognized that wall paintings may be deteriorated by

microorganisms as established by several studies that linked the observed damage with the

microbial community present as reviewed by Ciferri (1999) Despite this knowledge a

reliable characterization of bacteria associated with such damage is often lacking To

obtain greater insight into the bacterial community involved in biodeterioration of mural

paintings three sites (from the necropolis at Carmona Spain and the Saint-Catherine

chapel of the castle at Herberstein Austria) were sampled In previous studies from these

sites 385 isolates were analysed by fatty acid methyl ester GC and representatives of

different fatty acid clusters were further studied by 16S rRNA gene sequencing

(Heyrman and Swings 2001) Several of these representative strains showed the highest

sequence similarity with members of the genus Arthrobacter (Heyrman et al 2005)

Jimenez and Samson (1981) analysed the microbial flora of a large fresco painted

in the late 1920s in the monastery ldquoSanta Maria de la Rabidardquo (Huelva Spain) exhibiting

two types of deterioration white efflorescence and green-to-black stains




Tate and Rebrikova (1990) thought autotrophic nitrifying bacteria to be the first colonisers

of damaged frescoes and building materials in the cathedral of the nativity of the Virgin in

the Pafnutii-Borovskii monastery (Russia) The bulk of research on biodeterioration of

mural paintings has focused on explaining the observed phenomena by analysing the

microflora present Following this approach a precise characterisation of the

microorganisms using a polyphasic approach was never the first goal

Only a few studies included genomic fingerprinting (16S rDNA sequence analysis)

for the characterisation of bacteria associated with damage on mural paintings

[Rolleke (1996) Pinar et al (2001)] Previous studies based on culture-dependent

techniques showed that the flora present on frescos are fungi (predominant species of

Penicillium Aspergillus Cladosporium and Engyodontium) (Guglielminetti et al 1994)

bacteria (predominant species of Bacillus Arthrobacter Micrococcus Streptomyces and

Pseudomonas) (Heyrman et al 1999) cyanobacteria and eukaryotic algae (species of

Nostoc Lyngbya and Chlorophyceae) (Arino et al 1996 Calvo et al 1993) The killing

effect of an antimicrobial agent on a microorganism can be assessed in several ways

(1) Lethal activity may be expressed as the rate of killing by a fixed concentration

of drug under controlled conditions This rate is determined by measuring the number of

viable bacteria at various time intervals The resulting graphic depiction is known as the

time-kill curve Bacterial killing rates are in part dependent on the class of antibiotic and

the concentration of this agent With certain classes of antibiotics (eg aminoglycosides

and fluoroquino-lones) the rate of killing increases with increased drug concentrations up

to a point of maximum effect This is termed concentration-dependent bactericidal activity

In contrast the killing rates of szlig-lactam agents and vancomycin are relatively slow and

continue only as long as the concentrations are in excess of the MIC This rate of killing is

termed time-dependent bactericidal activity

(2) The minimal concentration of drug needed to kill most of the viable organisms

after incubation for a fixed length of time (generally 24 hours) under a given set of

conditions is the most common estimation of bactericidal activity and is known as either

the minimal bactericidal concentration (MBC) or the minimal lethal concentration (MLC)

It will be referred to in this document as the MBC Unfortunately the definition of the

MBC (999 killing of the final inoculum) is somewhat arbitrary and separates the

bacteria into two populations ndash a segrega-tion which might not have biological relevance




The determination of the MBC moreover is so subject to methodologic variables that the

clinical relevance of MBCs is nearly impossible to assess particularly for certain pathogen

and drug combinations (eg Staphylococci and beta-lactam agents)

(3) The serum of a patient receiving an antibiotic may be tested against the

infecting microorganism This can be done using time-kill curve methodology (ie serum

bactericidal rate) or using dilution methodology (ie serum bactericidal titer) The

principles of these methods as well as the influence of biological and technical factors are

similar (Barry et al 1999)

213 Microbial flora associated with deteriorated mural paintings

A great deal of research has been done in this field mainly by using cultivation dependent

methods However by using culture-dependent methods only a very small proportion of

microorganisms can be obtained Far less than 1 of organisms in the environment can be

cultivated by standard cultivation techniques (Pace 1996) Recent studies have shown that

culture-dependent techniques yielded mainly sporeforming bacteria suggesting that plating

leads to an over estimation of the number of spore-forming bacteria A wide number of

investigations have shown that in addition to chemical and physical processes

microorganisms play a major role in degradation of objects of art such as paintings stone-

works wood paper masonry leather parchment glass and metal (Ripka 2005)

A) Bacteria

In general growth of diverse organisms on mural paintings is supported by

humidity slight alkaline pH values and the presence of organic and inorganic nutrient

sources (Weirich 1989) In mural paintings pigments are suspended in water or oil often

binders such as casein and milk are used and they are supplied on the damp lime plaster

(Ciferri 1999) The calcium carbonate formed on contact with air consolidates the

pigments Thus frescos contain mainly inorganic components While chemoautotrophic

nitrifiers and thiobacilli obtain their energy for growth by oxidizing ammonium and

reducing sulphur-compounds respectively heterotrophic bacteria and fungi depend on

organically bound carbon [Tate et al (1991) Lazar (1971) Petushkova et al (1986)

Jimenez et al (1981)] Thus primary colonization of photoautotrophic Cyanobacteria

chemolithoautotrophic nitrifying bacteria and sulphuric acid producing thiobacilli represent




the first supply of organic matter The death and lysis of such bacteria promotes the

secondary settlement of heterotrophic bacteria and fungal populations (Calvo et al 1991)

Bacteria may derive from plants or pollen as many sequences obtained in this study

gave high score similarities with bacteria isolated from plants like Moraxella osloensis

Enterobacter cloace Enterobacter agglomerans and Pantoea endophytica Bacteria may

also derive from animal faeces like flies Bacillus Pseudomonas Aeromonas and Panteoa

(Ripka 2005) As Lazar and Dumitru (1973) reported studying mural paintings in a

Romanian monastery the genera Bacillus Arthrobacter and Pseudomonas have been

proved to produce salt efflorescences and were even considered to cause the blanching

phenomenon of frescoes (Jimenez et al 2000) Predominant bacterial genera are Bacillus

Streptomyces Arthrobacter Micrococcus and Pseudomonas [Weirich (1989)

Tate et al (1990) Arroyo et al (1997) Gonzalez et al (1999) Groth et al (1999)

Jimenez et al (2000)]

B) Fungi

It may be concluded that there is wide variety of fungal forms present on mural

paintings but Aspergillus and Alternaria were found dominating from all the experimental

sites next to them were Cladosporium Drechestera Fusarium Curvularia Chaetomium

Memnoniella Mycelia sterelai Pencillium Emericella Stachybotrytis Trichoderma

Phoma Monodictys Humicola and Epicoccum (Garg et al 1995)

The fungal genera most commonly found in lists of isolated taxa given in literature

are Alternaria Aspergillus Chaetomium Cladosporium Fusarium Penicillium and

Verticullum [Weirich (1989) Tate et al (1990) Guglielminetti et al (1994)

Garg et al (1995) Berner et al (1997) Ciferri (1999)] Some fungi having hard fruiting

bodies represented by sclerotia pycnidia perithecia cleistothecia and stroma could

organize themselves inside the painted layer Many fungal forms having such structures

were identified from our experimental sites viz Emericella nidulans A flavus

Chaetomium sp and Phoma sp etc These structures with little increase in humidity may

cause swelling in the paint film (Garg 1995)

Strains of species of Alternaria and Cladosporium both of them identified on the

mural paintings of the Palace of Schoumlnbrunn are the most common genera isolated from

stone and mural paintings in indoor environments with standard cultivation methods




(Ventosa 2004) Saprophytic species of the genus Cladosporium are among the most

abundant fungi isolated in saltern samples Commonly isolated Cyanobacteria belong to

Nostoc and eukaryotic algae to Lyngbya and members of the Chlorophyceae such as

Chlorella Pseudococcomyxa and Pseudopleurococcus [Gorbushina et al (1987)

Calvo et al (1993) Arino et al (1996)]

They grow optimally on media with low water activity due to high salt or sugar

concentrations seem to propagate 70 vegetatively in the salterns and are thus

halotolerantosmotolerant species rather than simple flush-through occurrences

Cladiosporum herbarum was frequently isolated from hypersaline waters dead and dying

plant material air soil food fruit cereals paint and textiles (Ventosa 2004)

C) Algae

Most of the painted areas in temperature climate became covered with green algae

(Pleurococcus Protococcus and Trentopholia) while under tropical conditions blue green

algae of the genera Oscillatoria and Sytonema predominated (Garg 1995) The main

factors which contribute to the growth of algae on wall paintings are the presence of

moisture in the plaster as well as availability of visible light These conditions are quite

prevalent in the tropical climate but in temperature regions also algal growth on wall

paintings have been reported [Tonolo et al (1961) Pietrini et al (1991)]

D) Actinomycetes

Usually literature refers to specific aspects eg the occurrence of cyanobacteria

and algae in caves with natural openings to sunlight or those with artificial lighting fungi

and so forth Actinomycetes have been considered to be mainly confined to soils where

they find their natural habitat but despite intensive studies there are still many gaps in our

knowledge of the role played by actinomycetes in soil processes and biodegrations of

murals cave paintings (Groth et al 1999) In addition halophilic bacteria actinomycetes

and cyanobacteria are also capable of growing on salt-affected mural paintings

(Karbowska 2003)

Monte and Ferrari (1993) reviewed the mechanisms of biodeterioration in tombs

and underground churches The role of actinomycetes in the deterioration of frescoes in

hypogean environments was stressed by Agarossi et al (1985) Monte and Ferrari (1993)

Giacobini et al (1987) described the serious damage suffered by a whole series of famous




frescoes in underground environments Actinomycetes were isolated from these mural

paintings located in crypts caves and tombs About 200 strains of actinomycetes were

isolated 60 of which were identified The most frequent species were Streptomyces albus

S cinereoruber S griseolus S vinaceus and some members of the Nocardia genus

Heterotrophic bacteria and actinomycetes were the most abundant microorganisms

on the frescoes of Etruscan tombs (Agarossi et al 1985) Bettini et al (1977) studied

some of the oldest Etruscan tombs (7th to 4th century BC) and found white-grey patinas

and spots from streptomycetes in the mural paintings They related the biodeterioration to

the microclimatic conditions (eg lower temperatures high RH) Monte and Ferrari (1993)

considered that colonization of the mural paintings was the result of selective competitive

and inhibitory dynamics among different microorganisms under different and changing

environmental conditions The pigments excreted by some actinomycetes were responsible

for the violet stains found on the walls of tombs in Tarquinia Similar stains were observed

in one of the Roman tombs the Circular Mausoleum the Necropolis of Carmona Spain

from which a Streptomyces sp producing violet pigments in culture media was isolated

214 Identification of biodeteriogens

The possibility of pigments and biodeteriogens identification in frescos and stone

monitoring is offered mostly by vibrational spectroscopy infrared thermography laser

induced fluorescence (LIF) laser induced breakdown spectroscopy (LIBS) XRF (X-Ray

Fluorescence) IR and UV emission Among them laser based methods allowing for in

situ or remote characterization of artwork surface were applied successfully for

prospecting diagnosis and maintenance (Colao et al 2005) The rep-PCR fingerprinting

of the isolates allowed determination of their genotypic diversity Studies that have

compared rep-PCR genomic fingerprint analysis with DNAndashDNA relatedness studies have

shown that the two techniques yield results that are in close agreement [Nick et al (1999)

Rademaker et al (2000)] These studies therefore suggest that rep-PCR fingerprinting can

be used as a genomic screening method to differentiate at the species level and to select

representatives for DNAndashDNA reassociation experiments (Heymann et al 2003)

The characterisation of bacteria was for example often limited to isolation from

group-specific media followed by microscopy [Sorlini et al (1987) Weirich (1989)

Tate et al (1990) and in more recent studies to morphological and physiological




characteristics including miniaturised and automated systems such as API and BIOLOG

[Arroyo et al (1997) Jimmenez et al (2000) and Laiz et al (2000)] or to

chemotaxonomic characteristics [Gonzalez et al (1999) Groth et al (1999)


Wagner et al (1994) demonstrated that conventional microbiological efforts can

fail to isolate all microorganisms present in natural samples DNA-based techniques used

to identify microorganisms have revealed only a smaller fraction of bacteria isolated from

artworks (Ward et al 1990 Rolleke et al 1996) Bacterial identification using molecular

techniques especially those including the sequencing of genes coding for ribosomal

16S rRNA is very important to study bacterial communities found on artefacts One such

method is Denaturing Gradient Gel Electrophoresis (DGGE) of PCR-amplified gene

fragments coding for rRNA (Muyzer et al 1993) that allows the separation of partial

16S rDNA amplified fragments of identical length but different sequence due to their

different melting behaviour in a gel system containing a gradient of denaturants

(Jimenez 2003)

Total genomic DNA was purified for 16S rRNA gene sequencing and rep-PCR

using a slight modification of the method of Pitcher et al (1989) as described by

Heyndrickx et al (1996) rep-PCR genomic fingerprinting was performed with the REP-

and (GTG)5 primers (Versalovic et al 1994) using the PCR conditions described

previously by Rademaker et al (1997) Electrophoresis and pattern analysis were further

performed as described by Heyrman et al (2003) 16S rRNA gene sequencing and

phylogenetic analysis were performed as described by Heyrman and Swings (2001)

Phylogenetic trees were constructed using the BioNumerics software (Applied Maths) by

applying the neighbour-joining and maximum-parsimony methods on a multiple alignment

similarity matrix The stability of relationships was assessed by a bootstrap analysis of

1000 datasets For determination of the G+C content and levels of DNAndashDNA

hybridization approximately 1 g biomass was harvested from agar plates DNA was

purified by using a combination of the protocols of Marmur (1961) and

Pitcher et al (1989) as described by Logan et al (2000) The G+C content of the DNA

was determined by HPLC (Mesbah et al 1989) using the further specifications given by

Logan et al (2000) DNAndashDNA hybridization experiments were performed using a




modification of the microplate method of Ezaki et al (1989) as described by

Willems et al (2001)

215 Preservation and conservation

Nowadays the conservation and preservation of our cultural heritage is one of the

world main concerns particularly in Europe The increasing need for non-destructive

investigation tools has become a major issue as sampling is in most cases restricted in

view of the value or the uniqueness of the object Artworks as wall paintings in buildings

and monuments of historical interest are affected by environment physical (temperature

and humidity) and biological factors (biodeteriogens) and by human (pollutant releases)

impact and therefore modified over the time The term ldquoconservationrdquo may have many

different meanings in different fields Every intervention directed to the restoration and

safeguard of works of art is commonly considered as a conservation treatment For

example conservation can involve the cleaning procedures set up by chemists or the photo

documentation of the preservation of historical memory For the appropriate conservation

of works of art even the lighting systems in a museum or the control of the visitorrsquos

movements in a site should be monitored Chemical degradation which induces the flaking

of paintings and deterioration from rain wind dust and other environmental causes is

mainly responsible for the weakening of the porous structure and the surface layers of the

materials used in works of cultural heritage (Colao et al 2005)

216 Conservation history

Wall paintings are an integral part of monuments and sites and should be preserved

in situ Many of the problems affecting wall paintings are linked to the poor condition of

the building or structure its improper use lack of maintenance frequent repairs and

alterations Also frequent restorations unnecessary uncovering and use of inappropriate

methods and materials can result in irreparable damage Substandard and inadequate

practices and professional qualifications have led to unfortunate results It is for this reason

that an appropriate document covering the principles of proper conservation-restoration of

wall paintings is necessary

The Venice Charter (1964) has provided general principles for the conservation-

restoration of cultural heritage The Amsterdam Declaration (1975) introducing the

concept of integrated conservation and the Nara Document on Authenticity (1994) dealing




with cultural diversity have expanded these principles Taking into account these and

additional relevant contributions such as the ICOM-CC Code of Ethics (1984) Document

of Pavia (1997) and ECCO Professional Guidelines (1997) the aim of this document is

to provide more specific principles for the protection preservation and the conservation

restoration of wall paintings This document therefore reflects basic and universally

applicable principles and practices and does not take into account particular problems of

regions or countries which can be supplemented at regional and national level by

providing further recommendations where necessary

217 Preservation techniques of the mural paintings

The removal of fat salts varnishes and pollutants from the surfaces of the works

of art are mostly done by chemical and mechanical methods At the end of the 19th

century the practices of restoration methods that do not inhibit further future interventions

was considered acceptable During the 1960s several chemicals such as polymeric resins

were synthesized and applied as a consolidation or protection agents The accepted idea

was that these substances could be removed at any time leaving a completely unaltered

substrate On the basis of origin archaeological objects can be classified as follows

(Carretti 2005)

1 Inorganic It includes siliceous and nonsiliceous materials Archaeological materials

produced from stones terracotta ceramics and glass can be included under siliceous

materials whereas objects made of metals can be included under nonsiliceous inorganic

materials Again metal objects can be classified as ferrous metals and non-ferrous metals

Objects made of iron are termed as ferrous metal objects whereas metal objects made from

other than iron is termed as non-ferrous metal objects

2 Organic It includes archival and anthropological materials Again archival materials

include paper bhojapatra tadapatra textiles wooden objects miniature paintings whereas

anthropological objects include bone and ivory leather feather and fur etc

In the past restoration of paintings and other old artwork often involved application

of acrylic resins to consolidate and protect them One of the most important tasks for

modern restorers is thus to remove these layers because it turns out that acrylic resins not

only by drastically changing the optics of the treated artwork but in many cases they

accelerate their degradation The scientists use an oil-in-water microemulsion with the




organic solvent para-xylene as the oil component The matrix makes the cleaning agent

viscous so that it cannot enter very far into the pores of a painting (Carretti 2005)

Nanoparticle treatment is the logical evolution of the Ferroni-Dini method

Dispersions of kinetically stable Ca (OH)2 nanoparticles in non-aqueous solvents solved

most of the drawbacks of the microsized powders Stable dispersions of calcium hydroxide

have been successfully applied (replacing polymers) as fixatives to re-adhere lifted paint

layers during many restoration workshops in Italy and in Europe as a consolidant The

FerronindashDini method (two steps the application of a saturated solution of ammonium

carbonate (NH4)2CO3 and the treatment with a barium hydroxide solution Ba(OH)2 also

called the barium method has long been the acknowledged method for the removal of

salts that threaten paintings reinforcing at the same time the porous structure However

commercially available carbonates and hydroxide powders have dimensions of several

micrometres much larger than the pores on the paint surface This means they donrsquot

penetrate the painting well and there is also a risk of damaging the artwork by a white

glaze forming on the surface (Berger 2006) Restoration should provide the reinforcement

of the porous structure and the consolidation of the surface layer of arte facts A few

simple principles can be considered to define the most appropriate restoration method

1) The treatment should be reversible so that one can revert to the original status of the

work of art at any desired time

2) All the applied chemicals must ensure the maximum durability and the chemical

inertness

3) The applied chemicals must invert the degradation processes without altering the

chemical composition of the artefacts and their physico-chemical and mechanical

properties (Baglioni 2006)

218 Commercial preservative used for preservation of mural paintings

Cleaning was done using soft brushes vacuum cleaner and Gumma paine (dough)

a natural latex sponge Areas near the window were very dirty with black soot and were

cleaned with micro fibrous cloths moistened with synperonic N (surfactant alkyl phenol

ethoxylate based on nonyl phenol) and neutralized with clean water In several places the

paint layers had separated from the substrate (exfoli-ation) In that cases loose paint flakes

were stuck back using Paraloid B72 (10 solution in ethanol) as sparingly as possible




Paraloid was injected with syringe behind the exfoliated area twice pushed and held back

with Melinex (polyester film) until fixed on the third application (Bionda et al 2002)

In a period of significant growth of molds and intensive efforts to stop the mold

contamination some of the lines of certain paintings were obscured or faded Solutions of

ethanol formalin 91 and Thiabendazole (TBZ) were used but they were not very

effective Para-formaldehyde fumigation was adopted beginning in 1981 to establish an

emergency treatment sensitivity of microorganisms from studied samples was tested

against Desogen by adding 1-3 (v v) in serial dilutions The best concentration

3 (vv) was applied with a brush to prevent spreading of the spores In order to overcome

the problem of fungal attack suitable fungicides carbendizeme or kiroxy-I or 5 solute of

N formaldehyde can be used Shellac solution in spirit or 2 solution of polyvinaycletate

in toluene can be used as preservative and consolidantsTo control insect and fungal attack

fumigants like pyrethrum DDT or 5 solution of sodium penta chlorophenate in kerosene

oil 2 to 5 solution of PVA in toluene or 2 to 5 solution of paraloid B-72 in toluene or

acetone be used (Miura 2009) Microclimatic conditions constitute a critical factor in the

conservation of cave paintings The unstable microclimatic condition is one of the factors

that always affect the state of conservation of mural paintings In the environmental-

monument system knowledge of the functional environment and climatic conditions of the

surroundings are necessary for the identification of causes of decay (Singh 2011)

Furthermore the risk of formation of indoor bioaerosols in residential and public

buildings (offices schools museums etc) has attracted interest in the last years due to

socio-economic as well as sanitary reasons (Ripka 2005) Thus it is important in aspects of

health and maintenance of Cultural Heritage to analyse the complete microflora to further

develop biocides for harmful microorganisms and methods for restoration of damaged

objects of art The biocidal testing was carried out on one representative algal species of

the two classes of algae viz Chlorococcum sp (green) and Chroococcus sp (blue-green)

found to be common in all the samples Efficiency tests were carried out both in liquid as

well as solid media containing algal suspensions by adding a biocide solution of known

concentration and determining the growth of the microorganism after the incubation

period The biocides used for the past studies were




1) Benzalkonium chlorides (MS Loba-chime Indo-Australian Co Bombay)

2) 2-Chloro-4 ethyl amino-6-isopropylamino-s-triazine commercially known as

Atrazine (MS Rallis India Ltd Bombay)

3) chloro-4 6-bis (ethylamino)-s-triazine commercially known as Simazine (MS

Bharat Pulverising Mills Ltd Bombay)

4) [3-(34-dichlorophenyl)-11-dimethyl urea] commercially known as Diuron (MS


Traditional conservators confront a challenging set of issues that must be resolved

prior to the restoration of any community mural In some cases the special nature of

community murals creates conflicts with conservatorsrsquo normal practices so it is useful to

begin by considering briefly the differences between community murals and other forms of

painting To do that it is necessary to better understand the nature of community murals

The biofilm was removed where possible with a low concentration of hydrogen peroxide

solution then the area was treated with about 70 isopropyl alcohol The conservation

techniques in Italy are barium hydroxide starts a reaction which converts the calcium

carbonate (lime) into calcium hydroxide (slaked lime) which means a new carbonation

process will take place and create adhesion to the wall The ammonium and water

evaporates The crystals of barium sulfate act as filler in the pores of the mural and can

also be washed away (Garreau 2007)

Another choice for protecting the mural is to use some of the various graffiti-

resistant finishes that are commercially available These range from protective wax

coatings that are removed with hot water to the 2-component solvent-based polyurethane

coatings Breath ability is important to the successful adhesion of acrylic products If a

mural encompasses an entire wall made of a masonry product it is advisable to apply thin

coats of MSA Varnish This will allow interior condensation and evaporating solvents and

out gassings to escape A mixture of shellac and alcohol in ratio of 5050 is also used

Readhesion of the paint layer was done by application of acrylic resin emulsion (primal

AC 335)the emulsion was applied by brush along the border with water and alcohol

(Agrawal and Pathak 2001)




219 Conservation agents

Indian Association for Study of Conservation of Cultural Property IASC was

founded in the year 1966 as a spin off of the UNESCOs Regional Seminar at New Delhi

The Association provided a National forum for the conservators practicing in isolation to

come together and share their experience among the peers and also to seek answers to

their problems and keep the professionals informed of the difficulties in carrying out their

work The forum took the responsibility of information dissemination about the new

developments in the area of conservation Formation of such a specialized professional

Association gave a big boost to the conservation research activity in the country

(Singh 1998)

The Archaeological Survey of India (ASI) under the Ministry of Culture is the

premier organization for the archaeological researches and protection of the cultural

heritage of India Maintenance of ancient monuments and archaeological sites and remains

of national importance is the prime concern of the ASI For the maintenance of ancient

monuments and archaeological sites and remains of national importance the entire country

is divided into 24 Circles The Archaeological Survey of Indiarsquos Science Branch is

responsible mainly for the chemical conservation treatment and preservation of some three

thousand five hundred ninety three protected monuments besides chemical preservation of

museum and excavated objects countrywide

Nanotechnology has recently found practical applications in the Conservation and

Restoration of the Worldrsquos Cultural Heritage (CAR-WHC) Baglionis group was among

the first to synthesize nanoparticles in non-aqueous solvents with the optimal properties for

application to cultural heritage conservation Nanoparticles of calcium and magnesium

hydroxide and carbonate have been used to restore and protect wall paints such as Maya

paintings in Mexico or 15th century Italian masterpieces (Berger 2006)

A few years back the INTACH Indian Council of Conservation Institutes (ICCI)

conducted a survey of wall paintings in various state of India and found that a large

number of wall paintings exist in temples palaces havelies churches forts and so on The

number of wall paintings in each state is very large but the researchers could locate and

examine only a few of them From some wall-paintings they collected samples of plasters

and paints and analyzed them They could get funding for conservation of some of them




and they were conserved by their teams (Agrawal 2007) A state-wise break-up of their

survey work is as given below table

Table-22 Status of Indian mural paintings (Agrawal 2007)

SNo Name of the State No of murals sites

surveyed

Status of sites

conserved

1 Bihar 8 Nil

2 Himachal Pradesh 4 2

3 Jammu amp Kashmir 17 Nil

4 Tamil Nadu 3 2

5 Madhya Pradesh 36 8

6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil

13 Uttar Pradesh 41 4

14 Uttarakhand 11 1

15 Orissa 79 5

220 Recent scenario of preservation

The scanning lidar fluorosensor experimental apparatus was designed by keeping in

mind that very often artworks and especially frescos on tombs are placed in small

chambers with a narrow entrance where only a limited space is available In order to

facilitate field operations the optical and mechanical supports have been assembled on a

small size optical bench contained in a box for easy transportation The compact LIF

system is designed to be coupled with another laser scanning device used for

characterization of surfaces the amplitude modulated LFR developed in the same

laboratory for 3D model reconstruction of object and environments (eg whole chambers)




The mechanical mountings are then conceived to offer a rapid and stable hookup to the

LRF apparatus in the case of simultaneous operation (Giovagnoli 2008)

221 Data recording

In order to facilitate the input and retrieval of data from the text fields a guide for

the compilation of the data record card was drawn up For each entry the guide explains the

type of data considered essential the technical vocabulary to adopt and cites a series of

illustrative examples showing the type of syntax and punctuation required The program

used was File Maker Pro 71 produced by the Claris Corporation The data record card is

divided up into six sections

1 Reference data This contains basic information relating to the work in question

(geographical location description of objectsite measurements author period

etc) The section ends with a paragraph on the methods of inspecting the painting

(visual tactile examinable support) which is of fundamental importance for

assessing the integrity and completeness of the survey (Giovagnoli 2008)

2 Documentation This section contains the necessary documentary references

relating to the work of a graphic photographic or written (bibliographical or

archival) nature

3 Positiondisplay characteristics The information contained in this section is brief

and acquired through simple visual examination as it will be presented in greater

detail on the environmental data record card

4 Technical data and conservation condition This is the most technical and analytical

section of the sheet in that it presents all the visually deducible data on the

constituent materials execution techniques and conservation condition of the

painting The section is divided into three parts presenting data that refers to the

individual layers or elements ideally comprising the work support structure

preparatory layers and paint layers The collection of data for each individual

section is carried out in the same order First the constituent materials and

execution techniques are examined and then the alterations linked to factors of

deterioration of a physico-chemical and biological nature and lastly the restoration

treatments carried out in the past An ldquoOtherrdquo entry is always included to provide

for those less common aspects not specified in the other fields As far as the




indicated factors of alteration are concerned an assessment of the extent of the

damage is recorded and a calculation expressed in percentage of its distribution in

relation to the overall surface (Giovagnoli 2008)

5 Indication of treatment programmes Based on the previously recorded

documentary technical and conservation elements this section proposes the

treatment programmes that need to be carried out indicating the type and degree of

urgency Five different time related categories were specified for this purpose

bull Emergency treatment

bull Short-term treatment

bull Medium-term treatment

bull Long-term treatment

bull Maintenance and inspection

222 Conclusion

From the above supporting articles the present study was designed in such a way to

develop a natural preservative gel for the conservation of the mural painting The present

study focuses on development of bio-based natural preservative coating for mural painting

and monuments The microbial flora of air from selected archeological sites was studied

The dye pigment from the plant sources and mineral sources used in traditional mural

painting were extracted and their antimicrobial property was studied The physico ndash

chemical characteristics of the extracted dyes were studied using SEM-EDAX FTIR and

GC-MS analysis Various combinations of these dyes and their antibacterial property were

studied Field trials for assessing the antimicrobial durability of developed natural

preservative were studied by coating it on mural paintings at Institute of Mural Paintings

Guruvayoor Kerala




11th


17th

and 19th

century AD

(Sasibhooshan 2006)



In south India they are found at Mehaboob Nagar (Andhrapradesh) Edakkal Marayoor

and Perimkadavila (Kerala) Mallampadi (Tamilnadu) and Bellary (Karnataka)These

paintings were considered as the early models of mural paintings The mural painting of


Style colour theme and technique of mural painting varied with different dynasties their

ruling period and important painting sites are given belowThe important mural painting

site Ajantha cave 9 amp10 was made during the ruling period from 2nd

BC to 2nd

AD of

Satavahana dynasty Vaishnava cave in Badami in the ruling period from 6th

-8th

AD

belongs to early western Chalukyas During 7th

-9th

AD Pallava dynasty in Kailasanatha

temple Kancheepuram was considered to be as one of the dominant mural painting site in

Southern India Similarly other South Indian dynasties early Pandya between 7th

and

9th

AD and early Chera from 8th

to 9th

AD highlighted their mural paintings at

Sittannavasal cave temple and Thirunandikkara cave temple respectively Mural paintitngs

at Kailasanatha temple Ellora from 8th

- 10th

AD was made by the dynasties of

Rashtrakuta The familiar mural paintings at Brihadeeswara temple Thanjavur and

Vijayalaya Choliswaram temple Narthamalai was carried out by the chola dynasty

between the period of 9th

and 13th

AD Another biggest dynasty called Vijayanagara

betweeen 14th

and 17th

AD made numerous mural paintings at Vidharbha temple

Lepakshi Varadaraja Perumal temple Kancheepuram Vitthala temple Hampi and Sri

Ranganatha tenple Srirangam Nayakarsquos dynasty ruled inthe period from 17th to 18th AD

established the mural paintings in Kapardisvara temple Tiruvanjuli and later murals of

Thanjavur and Kancheepuram Finally medivial Kerala (blend of Hoysala Chalukya and

Vijayanagara paintings) in 16th

and 18th

AD carried out the mural paintings at Tripayar

temple Ettumanur temple Thiruvanchikulam temple Panayanarkavu temple

Mattancherry Padmanabhapuram and Krishnapuram palace (Sivaramamurthi 1994)









































23 Indian murals



century BC and

5th




belong to 9th


belong to 11th


17th and

19th



























































in India











(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala









































23 Indian murals



century BC and

5th




belong to 9th


belong to 11th


17th and

19th



























































in India











(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala



















23 Indian murals



century BC and

5th




belong to 9th


belong to 11th


17th and

19th



























































in India











(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala


























































in India











(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala


























in India











(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala














in India











(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala










(Sasibhoosan 1987)











































Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala



























Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala
















Palace






















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala





















































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala








































































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala















































































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala













































































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala










































































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala








































































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala





































































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala



































(Salonen 2001)

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala

























(Ripka 2005)













































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala



































































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala
































source



bacteria























itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala




















itself








































































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala

























































A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala






















A) Bacteria




























B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala

















B) Fungi































C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala














C) Algae








D) Actinomycetes








(Karbowska 2003)
























































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala




















































(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala


















(Jimenez 2003)






































































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala






















































(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala




















(Carretti 2005)






































inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala
























inertness
























































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala














































































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala













































(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala













(Singh 1998)





























surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala








surveyed

Status of sites

conserved

1 Bihar 8 Nil



4 Tamil Nadu 3 2


6 Maharashtra 1 1

7 Kerala 61 3

8 Rajasthan 72 18

9 Haryana 1 1

10 Punjab 2 Nil

11 Karnataka 18 4

12 Sikkim 1 Nil


14 Uttarakhand 11 1

15 Orissa 79 5















221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala






221 Data recording














archival) nature






























222 Conclusion











Guruvayoor Kerala
















222 Conclusion











Guruvayoor Kerala

Documents

2. REVIEW OF LITERATURE 2.1 Introductionshodhganga.inflibnet.ac.in/bitstream/10603/43421/9/09...Review of Literature Figure-2.3: Ajanta caves Figure-2.4: Mural Painting of Ajanta caves