UGC SPONSORED NATIONAL CONFERENCE ON ...

UGC SPONSORED NATIONAL CONFERENCE ON CURRENT PERSPECTIVES AND CHALLENGES IN BIODIVERSITY AND ITS CONSERVATION (NACPCB-15) 6-7th March 2015

PROCEEDINGS ISBN 978-93-5212-382-7

Organized by PG & RESEARCH DEPARTMENT OF BIOCHEMISTRY RAJAH SERFOJI GOVT.COLLEGE,(AUTONOMOUS), THANJAVUR – 613 005, TAMIL NADU, INDIA.

In Collaboration with Amphigene Research Laboratories, Thanjavur 1

Edited by PG & Research Dept of Biochemistry, Rajah Serfoji Govt.College (Autonomous) Thanjavur – 613 005 Tamil nadu, India

ISBN 978-93-5212-382-7

Printed by Surya Press, Old Bus Stand, Thanjavur- 613 005

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A STUDY OF ETHNO MEDICAL PRACTICES USED IN TREATING CAPTIVE ELEPHANTS V.SIVASHANKAR AND DRA.LOGAMADEVI Department of Botany, NGM College, Pollachi INTRODUCTION Herbal medicines are currently in demand and their popularity is increasing day by day. About 500 plants with medicinal use are mentioned in ancient literature and around 800 plants have been used in indigenous systems of medicine. India is a vast repository of medicinal plants that are used in traditional medical treatments (Chopra et al., 1956). The various indigenous systems such as Siddha, Ayurveda, Unani and Allopathy use several plant species to treat different ailments (Rabe and Staden, 1997). Plant derived drugs came into use in the modern medicine through the uses of plant material as indigenous cure in folklore or traditional systems of medicine. (EVM) is a holistic comprehension of the indigenous systems of animal health, their interpretation through western medicine and the development of effective and appropriate technologies

(McCorkle,

1986).

According

to

Mathias-Mundy and

McCorkle

(1989)

Ethnoveterinary medicine (EVM) deals with the folk beliefs, knowledge, skills, methods and practices pertaining to the health care of animals. EVM is in practice by majority of the farmers / animal caretakers in one or the other form and have been found more useful where modern veterinary techniques are unavailable, or too expensive or difficult for the caretakers to access (Wanyama, 1997a,b). Some time EVM has been misunderstood to be linked to the use of only herbs, however it has more to offer regarding information, practices, tools, technologies, believes, breeds and Human resources (MathiasMundy and McCorkle 1989, Martin et al., 2001).

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Elephants Elephants has played an important role in the cultural, economic and social life of Asia from millennia. However, it has been increasingly marginalized in the regions having elephants; there is now little demand for the traditional work done by elephants. The significance of elephants as beasts of burden and a cultural symbol continues to last even today. India harbours more than 50percent of the wild elephant population and about 20 percent of the captive elephant population of Asia (Bist et al.,2001). The Asian elephant (Elephas maximus) enjoys a special status in the country and the elephant symbolises the Indian ethos. It has been very closely associated with the religion, myths, history, and cultural heritage of India for centuries. Protecting and ensuring the survival of the elephant means much more to an Indian than protecting just another endangered species. Biology of Elephants The Asian elephant is one of the few extant mega herbivores (Owen-Smith, 1988). Males grow faster than female from the age of 2 years onwards (Sukumar et al., 1988). Adult bulls are typically 240–300 cm in height and 3500–6000 kg in body mass compared to 195–240 cm and 2000–3500 kg for adult cows, although there is considerable regional variation. Elephants are highly intelligent (Rensch, 1957) and have superior cognitive abilities. Domesticating Elephants India has a fascinating history of domesticating wild elephants. Lahiri Choudhury (1989) has traced, on the basis of rock paintings, the history of domesticated elephants in India to about 6000 B.C. Seals of the Indus Valley Civilization (2500 – 1500B.C) also suggest the presence of domesticated elephants in India at that time. The Ashokan edicts refer to the setting up of hospitals for the treatment of elephants and other animals (Bist et al.,2001).

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Mahout relationship Elephant is a big and intelligent animal. Taking care of such an animal is itself a strenuous job and also needs a lot of understanding and thinking on the part of a mahout. The daily routine of a mahout of a working elephant is also very demanding Mahouts should have stamina, quick reflexes, sense of discrimination, and most of all patience, perseverance and responsibility.Common sense and a sharp mind are essential qualities for a mahout to handle in a crisis. A clever mahout always use intelligence and clever tactics, as the elephant is a very intelligent creature (www.elephantcare.com). Ethno-veterinary medicine (EVM) Ethno-medicine is a sub-field of ethnobotany or medical anthropology that deals with the study of traditional medicines: not only those that have relevant written sources but especially those for which knowledge and practices have been orally transmitted over the centuries. . EVM or ethno-veterinary research was defined by McCorkle as: “The holistic, interdisciplinary study of local knowledge andits associated skills, practices, beliefs, practitioners, and social structures pertaining to the healthcare and healthful husbandry of food, work, and other income-producing animals, always with an eye to practical development applications within livestock production and livelihood systems, and with the ultimate goal of increasing human well-being via increased benefits fromstock raising.” (Martin et al., 2001) Ethno-veterinary medicines/remedies are practical in use, effective and cheap. They rely on local plants or easily available materials. They reflect centuries of experience of application and learning through trial-and-error (Jabbar et al., 2006).

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In most rural areas people prefer to treat their animals with indigenous drugs. At present over 35,000 plants are known to have healing properties (Jain,1991). All parts of the plants, including

leaves,

bark, fruits,

flowers, seeds

are used in

medicinal

preparations

(McCorkle,1996). Ethnoveterinary practices are often cheap, safe, time tested and based on local resources and strengths. These can provide useful alternatives to conventional animal health care (Kumar,2002). Ethnoveterinary medicine is in danger of extinction because of advancement of the modern veterinary medicine. The importance of the traditional knowledge on ethnoveterinary practices by specialists (Vaidyas) and local healers who are knowledgeable and experienced in traditional systems of treatment, but their knowledge is not documented, and is dwindling fast (Jain,2000). As the local healers did not document their knowledge and experience, and did not pass it on to others readily, there was danger of extinction of that knowledge (Mathias and Anjaria,1998). This study aims to collect such knowledge / information in treating captive elephants and documenting it in a proper format Objectives and methdology The purpose of the study is to document the indigenous/ ethnic knowledge of the mahouts, tribes and other people closely associated with elephants about caring and treating elephants with herbal remedies for common ailments and diseases. Following are the objectives of the current study. 

To collect the traditional knowledge related to elephant caring from the experienced mahouts/care takers.



To provide a detailed list of plant species and other caring practices for the treatment of ailments in captive elephants.

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

To document the information on utilization of medicinal plants for the treatment and to conserve the valuable but rapidly disappearing traditional knowledge of treating / caring of elephants.



To make available the collected data for the interested new generation, mahouts/ care takers of the pachyderms.

Description of Study Area The study areas include Mudumalai, Wayanad and Palakkad. These areas were selected on the basis of availability of mahouts/caretakers who are well experienced in handling the animal and who are ready to share the information’s. Period of visit Field trips were done to the above said areas in between December 2012 to January 2014 as a part of study and collection of data related to ethnomedical practices in captive elephants. Interview with mahouts/caretakers Data were collected from experienced mahouts according to the methodology suggested by Jain, 2001. Mahouts were selected on the basis of their working experiences (< 20years) with the elephants. A total of 8 mahouts were interviewed. The age of the mahouts was between 30 75 years. Questionnaire method was adopted for collecting information from mahouts. The questionnaire surveys were done by using a pre structured questionnaire. It was adjusted during the survey process. Data analysis The reported problems/ conditions were grouped into 13 categories on the basis of information gathered. 7



Impaction



Colic



Foot rot



Wounds



Wounds caused by using chains/ropes



Edema



Insect bite



For proper tusk growth



Skin problems



Body ache



Disturbances by ticks and flies



Eye Problem



Maggot infestation

There has been an avid interest in the study of medicinal plants and their traditional healing properties in veterinary practices by either tribal people or indigenous communities of India.The present study was taken up to ascertain the detailed information on the traditional practices in caring captive elephants. Results & Discussion Disorders/ Problems in captive elephants and the traditional ailments 

COLIC

Colic is a condition of abdominal disorder, commonly seen in elephants. It occurs due to irregularities in diet and water intake

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Traditional Practices 

A herbal mixture can be prepared. The ingredients are Asafoetida, jaggery , dried ginger ,garlic and chilies. They are altogether crushed into a paste and can be placed inside the concentrate food or elephant’s ration and can fed to the animals. The particular results can be compared with the results of Tyler 1994 &Spelman et al.,

2006. According to Tyler, Garlic is having antiseptic property which was first demonstrated by Louis Pasteur (Tyler 1994). Spelmen 2006 confirms the presence of allyl sulfides which are known for antitumour, anticarcinogenic, and antistress action. Asafoetida possesses antifungal, antiviral, anti – diabetic, anti – inflammatory and anti – mutagenic activities. It is also shown to be a remedy of great value as a carminative in flatulence and colic. It also exerts a significant anthelmintic action against round worms and pinworms. It has been found useful in ttreating neck sore, paralysis, stomach – ache and indigestion (Iranshahy and Iranshahi, 2011). Impaction- It is a condition of Lack of appetite. Animal does not eat fodder. Body posture suggests discomfort. Lays down and gets up frequent. Dung may be absent, fewer in numbers or smaller in size. Stomach may look bloated. Dull appearance and lack of interest in drinking water Traditional practices 

Hot compresses may be applied on the flanks and abdomen. A cloth compress of sand or fried sawdust (to provide dry heat) can be used, for all kinds of stomach ailment. It relieves pain.



During impaction, the mahouts/keepers lubricate their hands with oil, insert the hand into the rectum and try to relieve the impaction manually. 9



Foot rot

Fungal infection on the heel or soles and toes of an elephant are invariably due to unhygienic conditions of the tethering site and cause tremendous pain to the animal. Traditional Practices 

Apply the mixture prepared by boiling mustard oil mixed with crushed nuts of Bhela (washerman’s nut i.e. Semecarpus anacardium) and a little blue vitriol (copper sulphate) at the affected feet.



Elephants are not allowed to get the affected feet wet. The wound is treated with the mixture of turpentine oil, coconut oil and carbolic acid till it completely heals up. In Ayurvedha and Siddha (Indian systems of medicine) classics, copious references regarding the properties and uses of S.anacardium nuts are found (Sharma et al., 1966). The fruit of S.anacardium is acrid, hot, sweetish, edible aphrodisiac, anthelminthic,causes looseness of bowels, removes ascites, skin diseases, heals ulcers and is useful in treating asthma (Krithikar & Basu, 1975).

Conclusion& Recommendations In the present study, 31 medicinal plants used for treating different problems of elephants were recorded and documented. For more than a decade now ethno veterinary medicine has experienced a revival and several reports have been published. This growing interest in traditional practices had been encouraged by the recognition of some efficacious ethno veterinary medicinal products. Ethno veterinary medicine often provides cheaper options than comparable western drugs, and the products are locally available and more there is increasing interest in the field of ethno veterinary research and development easily accessible. In the face of these and other factors, In spite of rapid progress and spread of modern medicine and surgery, faith in and 10

popularity of traditional methods has not decreased in treating captive elephants. The importance of these traditional medicines / practices may realized worldwide as some of them may prove to be very effective and some other prescriptions of these traditional practioners may benefit to the entire elephant community when thorough scientific analysis is conducted. This study shows the social importance of the floristic richness, particularly regarding the significance of medicinal plants in treating/ caring captive elephants. This is reflected in the great diversity of plants used for medical purposes as well as in the wide range of their applications and associated procedures. Ethano veterinary explorations play vital role in bringing to light information about valuable plant species from our rich flora that can be source of safer and cheaper potent drugs for the benefit of entire elephant community. The traditional knowledge with its holistic and systems approach supported by an experimental base can serve as an innovative and powerful discovery engine for newer, safer and more affordable medicines. A great deal of information about the traditional uses of plants is still intact with the traditional caretakers. Plant derived medicines have greatly contributed to animal health and well-being, and today, plant materials have provided the models for, a significant proportion of the modern drugs. References 1. Bist, S.S., Cheeran, J.V, Choudhury, S., Barua, P. and Misra, M.K. 2001. The domesticated Asian elephant in India Giants on Our Hands, Proceedings of the International Workshop on the Domesticated Asian Elephant, Bangkok, Thailand, pp. 129–148. 2. Iranshahy, M. and M. Iranshahi, 2011. Traditional uses, Phytochemistry and pharmacology of asafetida (Ferula asafoetida): A review. J.ethanopharmacol. 134:1-10.

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3. Jabbar, A., Raza, M.A., Iqbal, Z. & Khan, M.N. (2006). An inventory of the ethnobotanicals used as anthelmintics in the southern Punjab, Pakistan. Journal of Ethnopharmacy, 108, 152-154.

4. Jain, S.K., 2000. Plants in Indian ethnoveterinary medicine: Status and Prospects. Ind. J. Vet. Med., 20: 1 - 11. 5. Jain,S.K. 1991.Dictionary of Indian folk medicine and etnobotany. Deep publications, New Delhi, India. 6. Kayode.J, M. K. Olanipekun and P. O. Tedela 2009. Medicobotanical Studies in Relation to Veterinary Medicine in Ekiti State, Nigeria: (1) Checklist of Botanicals used for the Treatment of Poultry Diseases. Ethnobotanical Leaflets 13: 40-46. 2009. 7. Kirti Sinha, N P Mishra, J Singh & S P S Khanuja 2004. Tinospora cordifolia, a reservoir plant for therapeutic applications: A review. Indian Journal of traditional Knowledge. 3(3). 257-270. 8. Krithikar KR & Basu BD. 1975. Indian Medicinal Plants. Vol 1. (MS Bishen Singh – Mahendra Paul Singh Publishers, Dehradun, India) 667. 9. Kumar, D. 2002. The use and relevance of ethnoveterinary practices in sheep. Ind. J. SmallRuminants, 8 (2): 124 - 128. 10. Lahiri choudhury, D.K. 1989. The Indian elephant in a changing world. In Carla M.Borde, ed.Contemporary Indian tradition: voices on culture, nature and the challenges of change. Edited by. Washington and London: Smithsonian institution Press.Lam.Int j Green Pharm 2008., 2:218-9.

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11. M.N.B.Nair 2006.- Documentation and assessment of ethnoveterinary practices from an ayurvedic viewpoint. Traditional knowledge systems of India & Srilanka, -77pp. 12. Mahima, Anu Rahal, rajib deb, Shyma K. Latheef, Hari Abdul, Ruchi Tiwari , Amit kumar varma, Amit Kumar and K.Dhama 2012. Immunomodulatory and therapeutic Potentials of Herbal, traditional Indigenous and Ethnoveterinary medicines. Pakistan Journal of Biological Sciences. 1-22. 13. Mathias, E., and Anjaria, J. 1998. Forthcoming Prospects of ethnoveterinary medicine in the modern world. In: Directory on Indian agriculture. Rashtra Deepika Ltd., Kerala, India. pp. 526 – 534. 14. Mathias-Mundy, E. and McCorkle, C. M. 1989. Ethnoveterinary Medicine: An annotated bibliography. Bibliographies in Technology and Social Change No. 6. Center for Indigenous Knowledge and Agricultural andRural Development (CIKARD). Iowa State University Research Foundation, Ames, Iowa, USA. 15. McCorkle, C. M. 1996. An Introduction to Ethnoveterinary Research and Development. J. of Ethnobiology,6 (1):129-149. 16. Misra B. 1966. - The Rajamartanda of Sri Bhojaraja with the 'Vidyotini' Hindi Commentary. Chowkhamba Vidyabhawan, Varanasi, 108-112. 17. Mukhopadhyaya G.N. 1926. - History of Indian Medicine, Vol. II. University of Calcutta, 356-424,400-424,491-495. 18. Owen-Smith, R. N. (1988): Megaherbivores: the influence of very large body size on ecology. Cambridge:Cambridge University Press.

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19. Pallavi Srivastavaa: and S. Durgaprasad 2008. Burn wound healing property of Cocos nucifera. An appraisal. Indian J Pharmacol. Aug 2008; 40(4): 144–146. 20. Pande PC, Lalit Tiwari & HC Pande. 2007. Ethano veterinary plants of Uttaranchal – A review. Indian journal of Traditional Knowledge. 6(3). 444-458

21. Rabe, T. and Staden, J. V. 1997. Antibacterial activity of South African plants used for medicinal purposes, J. Ethnopharmacol. 56: 81-87. 22. Raman Sukumar 2003.The Living Elephants: Evolutionary Ecology, Behavior, and Conservation. Oxford university press PP:77. 23. Rawat GS,Neem(Azadirachta indica) Natures drug store, Indian For, 1995,25,145-152. 24. Rensch, B. (1957): The intelligence of elephants. Scientific AmericanFebruary: 44–49. 25. Sharma P.V. 1983. - Caraka-Samhita (text with English translation), Vol. II. Chaukhambha Orientaba, Varanasi & Delhi, 665-666. 26. Smita jain and U K Patil. 2010. Pytochemical and pharmacological profile of Cassia tora Linn. – An Overview. Indian journal of natural Products and resources. 1(4). 430437. 27. Srikantha murthy K.R. 1987. - Medicine and Allied Sciences in Sivatattvaratnakara. Bull. Indian Inst. Hist. Med., 17, 89-93. 28. Sukumar, R. & Gadgil, M. (1988): Male–female differences in foraging of crops by Asian elephants. Animal Behaviour 36: 1233–1235.

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29. Tamal Mondal. 2012. An investigation on ethno-veterinary medicinal plants of Siliguri Subdivision of Darjeeling District, West Bengal, India. Journal of Today’s Biological Sciences : Research & Review (JTBSRR) ISSN 2320-1444 (Online)Vol.1,Issue 1,page 45-50. 30. Thatte, U.m. and S.A. Dhanukar, 1997. The Rasayan concept: Clues from Immunomodulatory Therapy. In: Immunomodulation, Upadhayaya, S.N.(Ed.) Narosa publishing House, New Delhi, PP 141-148. 31. Tyler, V.E., 1994. Herbs of Choice. The Therapeutic use of Phytomedicinals. Haworth Press, New York, Page: 209. 32. Wanyama, J. 1997a. Ethnoveterinary knowledge among pastoralists of Samburu, Kenya. ITDG/IIRR. 33. Wanyama, J. 1997b. Ethnoveterinary treatment study among the Samburu and Turkana communities in Bargoi. ITDG, Nairobi, Kenya. ITDG/OXFARM. 34. www.elephant care.com *****************************************

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INDIAN LEGISLATION ON BIODIVERSITY AND ITS ROLE IN CONSERVATION By R.Thiyagarajan, M.L., Lecturer in Law, The Central Law College, Salem and G. Arun, Vth Year B.A.B.L., The Central Law College, Salem.

INTRODUCTION

India is one of the 12 mega biodiversity countries of the world and one among the 194 signatories to the Convention on Biological Diversity at Earth Summit in Rio de Janeiro in 1992. With only 2.5% of the earth’s land area, India accounts for 8% of the recorded species of the world which includes millions of races, subspecies and local variants of species and the ecological processes and cycles that link organism into population, communities and all different ecosystems. Demographically, it is the second largest populated country in the world and a majority of its population directly depends on biological resources for livelihood. The necessity to protect bio-diversity also engaged the attention of the Supreme Court of India in Research Foundation for Science Technology and Ecology v. Ministry of Agriculture1. The Government was reminded to – 1. have legislation for the protection of bio-diversity, 2. the geographical indicators, 3. Plant varieties and farmers ‘rights’ protection. As a matter of fact for this purpose the government informed the Supreme Court of India that a Bill stood drafted. To give effect to that, The Biological Diversity Act was enacted by the Parliament of India in the year 20022 to attain the following objects: 1. To provide for conservation of biological diversity, 2. To provide for sustainable use of its components, and 3. To provide for fair and equitable sharing of the benefits arising out of the use of biological resources, knowledge, and 4. For matters connected therewith or incidental thereto.

1 2

(1999) 1 SCC 655. Act No.18 of 2003; received the assent of the President of India on the 5 th February 2003.

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Regulation of Access to Biological Diversity Section 3 of the Biological Diversity Act, provides that the following persons shall be required to obtain the approval of the National Biodiversity Authority for obtaining any biological resource occurring in India or knowledge associated thereto for research or for commercial utilization or for bio-survey and bio-utilization. (a) a person who is not a citizen of India; (b) a citizen of India, who is a non-resident as defined in clause (30) of Section 2 of the Income tax Act, 1961; (c) a body corporate, association of organization – (i)

not incorporated or registered in India; or

(ii)

Incorporated or registered in India under any law for the time being in force which has any non-Indian participation in its share capital or management.

Section 4 prohibits the transfer of results of any research relating to any biological resources occurring in, or obtained from, India for monetary consideration or otherwise to any person who is not a citizen of India or citizen of India who is non-resident as defined in clause (30) of Section 2 of the Income Tax Act, 1961 or a body corporate or organization which is not registered or incorporated in India or which has any non-Indian participation in its share capital or management without the previous approval of the National Biodiversity Authority. Application for Intellectual Property Rights Section 6 of the Biological Diversity Act provides that “No person shall apply for any intellectual property right, by whatever name called, in or outside India for any invention based on any research or information on a biological resource obtained from India without obtaining the previous approval of the National Biodiversity Authority before making such application.” However, if a person applies for a patent, permission of the National Biodiversity Authority may be obtained after the acceptance of the patent but before the seating of tile patent by the patent authority concerned.

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It is also provided that the National Biodiversity Authority shall dispose of the application for permission made to it within a period of ninety days from the date of receipt thereof. The National Biodiversity Authority may, while granting the approval under this Section, impose benefit sharing fee or royalty or both or impose conditions including the sharing of financial benefits arising out of the commercial utilization of such rights. Prior intimation to State Biodiversity Board for obtaining Biological Resource for certain purposes Section 7 of the Biological Diversity Act provides that “ No shall person who is a citizen of India, shall obtain any biological resource for commercial utilization, or bio-survey and bioutilization for commercial utilization except after giving prior intimation to the State Biodiversity for commercial utilization concerned. However the section exempts the local people and communities of the area, including growers and cultivators of biodiversity, and vaids and hakims, who have been practicing indigenous medicine from giving prior intimation to the State Biodiversity Board. Establishment of National Biodiversity Authority The Central Government shall establish the National Biodiversity Authority, having its head office at Chennai and the National Biodiversity Authority may, with the previous approval of the Central Government, establish officers at other places in India. The National Biodiversity Authority shall consist of the following members: (a) A chairperson, who shall be an eminent person having adequate knowledge and experience in the conservation and sustainable use of biological sustainable use of biological diversity and in matters relating to equitable sharing of benefits, to be appointed by the Central Government; (b) Three ex-officio members to be appointed by the Central Government, one representing the Ministry dealing with Tribal Affairs and two representing the Ministry dealing with Environment and Forests or the Director General of Forests;

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(c) Seven ex-officio members to be appointed by the Central Government to represent respectively the Ministries of the Central Government dealing with – 1. Agricultural Research and Education; 2. Biotechnology; 3. Ocean Development; 4. Agriculture and Cooperation; 5. Indian Systems of Medicine and Homoeopathy; 6. Science and Technology; 7. Scientific and Industrial Research; (d) Five non-official members to be appointed from amongst specialists and scientists having special knowledge of, or experience in, matters relating to conservation of biological diversity, sustainable use of biological resources and equitable sharing of benefits arising out of the use of biological resources, representatives of industry, conservers, creators and know ledge-holders of biological resources. Committees of National Biodiversity Authority The National Biodiversity Authority may constitute a committee to deal with agrobiodiversity. “Agro-biodiversity” means biological diversity, of agriculture related species and their wild relatives.3 The National Biodiversity Authority may also constitute such number of committees as it deems fit for the efficient discharge of its duties and performance of its functions under this Act. Functions and powers of the National Biodiversity Authority Section 18 of the Biological Diversity Act prescribes the functions and powers of the National Biodiversity. The following are the functions and powers of the National Biodiversity Authority: 1. It shall be the duty of the National Biodiversity Authority to regulate activities referred to in Section 3, 4 and 6 and by regulations issue guidelines for access to biological resources and for fair and equitable benefit sharing.

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Section 13 of the Biological Diversity Act, 2002.

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2. The National Biodiversity Authority may grant approval for undertaking any activity referred to in Section 3, 4 and 6. 3. The National Biodiversity Authority may – (a) Advise the Central Government on matters relating to the conservation of biodiversity, sustainable use of its components and equitable sharing of benefits arising out of the utilization of biological resources; (b) Advise the State Government in the selection of areas of biodiversity importance to be notified under sub-section (1) of section 37 as heritage sites and measures for the management of such heritage sites; (c) Perform such other functions as may be necessary to carry out the provisions of this Act. 4. The National Biodiversity Authority may, on behalf of the Central Government, take any measures necessary to oppose the grant of intellectual property rights in any country outside India on any biological resource obtained from India or knowledge associated with such biological resource which is derived from India. Transfer of Biological Resource or Knowledge4 No person who has been granted approval under Section 19 shall transfer any biological resource or knowledge associated thereto which is the subject matter of the said approval except with the permission of the National Biodiversity Authority. Any person who intends to transfer any biological resource or knowledge associated thereto shall make an application to the National Biodiversity Authority in such form and in such manner as may be prescribed. On receipt of an application under sub-section (2), the National Biodiversity Authority may, after making such enquiries as it may deem fit and if necessary after consulting an expert committee constituted for this purpose, by order, grant approval subject to such terms and conditions as it may deem fit, including the imposition of charges by way of royalty or for reasons to be recorded in writing, reject the application: provided that no such order for rejection shall be made without giving an opportunity of being heard to the person affected.

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Section 20 of the Biological Diversity Act, 2002.

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The National Biodiversity Authority shall give public notice of every approval granted by it under this section. Determination of equitable benefit sharing by National Biodiversity Authority The National Biodiversity Authority shall while granting approvals under Section 19 or Section 20 ensure that the terms and conditions subject to which approval is granted secures equitable sharing of benefits arising out of the use of accessed biological resources, their byproducts, innovations and practices associated with their use and applications and knowledge relating thereto in accordance with mutually agreed terms and conditions between the person applying for such approval, local bodies concerned and the benefit claimers. The National Biodiversity Authority shall, subject to any regulations made in this behalf, determine the benefit sharing which shall be given effect in all or any of the following manners, namely: 1. Grant of joint ownership of intellectual property rights to the National Biodiversity Authority, or where benefit claimers are identified, to such benefit claimers; 2. Transfer of technology; 3. Location of production, research and development units in such areas, which will facilitate better living standards to the benefit claimers; 4. Association of India scientists, benefit claimers and the local people with research and development in biological resources and bio-survey and bio-utilization; 5. Setting up of venture capital fund for aiding the cause of benefit claimers; 6. Payment of monetary compensation and non-monetary benefits to the benefit claimers as the National Biodiversity Authority may deem fit. Where any amount of money is ordered by way of benefit sharing, the National Biodiversity Authority may direct the amount to be deposited in the National Biodiversity Fund: provided that where biological resources or knowledge was a result of access from specific individual or group of individuals or organizations, the National Biodiversity Authority may direct that the amount shall be paid directly to such individual or group of individuals or organizations in accordance with the terms of any agreement and in such manner as it deems fit.

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For the purposed of this Section, the National Biodiversity Authority shall, in consultation with the Central Government, by regulations, frame guidelines. Establishment of State Biodiversity Board With effect from such date as the State Government may, by notification in the Official Gazette, appoint in this behalf, there shall be established by that Government for the purposes of this Act, a Board for the State to be known as the (name of the State) Biodiversity Board. In a union territory, the National Biodiversity Authority shall exercise the powers and perform the functions of a State Biodiversity Board for that Union territory: Provided that in relation to any Union territory, the National Biodiversity Authority may delegate all or any of its powers or functions under this sub-section to such person or group of persons as the Central Government may specify. The Board shall consist of the following members, namely: (a) A chairperson who shall be an eminent person having adequate knowledge and experience in the conservation and sustainable use of biological diversity and in matters relating to equitable sharing of benefits, to be appointed by the State Government; (b) Not more than five ex-officio members to be appointed by the State Government to represent the concerned Departments of the State Government; (c) Not more than five members to be appointed from amongst experts in matters relating to conservation of biological diversity, sustainable use of biological resources and equitable sharing of benefits arising out of the use of biological resources. The head office of the State Biodiversity Board shall be at such place as the State Government may, by notification in the Official Gazette, specify. Functions of State Biodiversity Board Section 23 of the Biological Diversity Act prescribes the functions of the State Biodiversity Board. The functions of the State Biodiversity Board are: 22

(a) Advice the State Government, subject to any guidelines issued by the Central Government, on matters relating to the conservation of biodiversity, sustainable use of its components and equitable sharing of the benefits arising out of the utilization of biological resources; (b) Regulate by granting of approvals or otherwise requests for commercial utilization or bio-survey and bio-utilization of any biological resource by Indians; (c) Perform such other functions as may be necessary to carry out the provisions of this Act or as may be prescribed by the State Government. Constitution of National Biodiversity Fund There shall be constituted a Fund to be called the National Biodiversity Fund and there shall be credited thereto(a) Any grants and loans made to the National Biodiversity Authority under Section 26; (b) All charges and royalties received by the National Biodiversity Authority under this Act; and (c) All sums received by the National Biodiversity Authority from such other sources as may be decided upon by the Central Government. The fund shall be applied for – (a) Channeling benefits to the benefit claimers; (b) Conservation and promotion of biological resources and development of areas from where such biological resources or knowledge associated there to has been accessed; (c) Socio-economic development of areas referred to in clause (b) in consultation with the local bodies concerned. Constitution of State Biodiversity Fund There shall be constituted a Fund to be the State Biodiversity Fund and there shall be credited thereto – (a) Any grants and loans made to the State Biodiversity Board under Section 31; (b) Any grants or loans made by the National Biodiversity Authority; 23

(c) All sums received by the State Biodiversity Board from such other sources as may be decided upon by the State Government. Biodiversity Heritage Sites5 Without prejudice to any other law for the time being in force, the State Government may, from time to time in consultation with the local bodies, notify in the Official Gazette, areas of biodiversity importance as biodiversity heritage sites under the Act. The State Government, in consultation with the Central Government, may frame rules for the management and conservation of all the heritage sites. The State Government shall frame schemes for compensating or rehabilitating any person or section of people economically affected by such notification. Power of Central Government to notify threatened species6 Without prejudice to the provisions of any other law for the time being in force, the Central Government, in consultation with the concerned State Government, may from time to time notify any species which is on the verge extinction or likely to become extinct in the near future as a threatened species and prohibit or regulate collection thereof for any purpose and take appropriate steps to rehabilitate and preserve those species. Constitution of Biodiversity Management Committee7 Every local body shall constitute a Biodiversity Management Committee within its area for the purpose of promoting conservation, sustainable use and documentation of biological diversity including preservation of habitats, conservation of land races, folk varieties and cultivars, domesticated stocks and breeds of animals and microorganisms and chronicling of knowledge relating to biological diversity. For the purposes of this sub-section, -

5

Section 37 of the Biological Diversity Act, 2002. Section 38 of the Biological Diversity Act, 2002. 7 Section 41 of the Biological Diversity Act, 2002. 6

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(a) “Cultivar” means a variety of plant that has originated and persisted under cultivation or was specifically bred for the purpose of cultivation; (b) “Folk variety” means a cultivated variety of plant that was developed, grown and exchanged informally among farmers; (c) “Landrace” means primitive cultivar that was grown by ancient farmers and their successors. The National Biodiversity Authority and the State Biodiversity Boards shall consult the Biodiversity Management Committees while taking any decision relating to the use of biological resources and knowledge associated with such resources occurring within the territorial jurisdiction of the Biodiversity Management Committee. Constitution of Local Biodiversity Fund8 There shall be constituted a Fund to be called the Local Biodiversity Fund at every area notified by the State Government where any institution of self-government is functioning and there shall be credited thereto(a) Any grants and loans made under Section 42; (b) Any grants or loans made by the National Biodiversity Authority; (c) Any grants or loans made by the State Biodiversity Boards; (d) Fees referred to in sub-section (3) of Section 41 received by the Biodiversity Management Committees; (e) All sums received by the Local Biodiversity Fund from such other sources as may be decided upon by the State Government. Penalties Section 55 of the Biological Diversity Act prescribes punishments for contravention of various provisions of the Act. The punishments prescribed are: Whoever contravenes or to or abets the contravention of the provisions of Section 3 or Section 4 or Section 6 shall be punishable with imprisonment for a term which may extend to

8

Section 43 of the Biological Diversity Act, 2002.

25

five years, or with fine which may extend to ten lakh rupees and where the damage caused exceeds ten lakh rupees such fine may commensurate with the damage caused, or with both. Whoever contravenes or attempts or contravene or abets the contravention of the provisions of Section 7 or any order made under sub-section (2) of Section 24 shall be punishable with imprisonment for a term which may extend to three years, or with fine which may extend to five lakh rupees, or with both. Conclusion The Biological Diversity Act, 2002 has been enacted to protect biological resources in India. The Supreme Court of India in State of Himachal Pradesh v. Ganesh wood products, 9 held that a decision making authority must give due weight and regard to ecological factors such as the environmental policy of government and the sustainable use of natural resources. A government decision that fails to take into account relevant considerations affecting the environment is invalid.10 From the above said cases, it is clearly stated that if anything goes wrong against biological resources or natural resources even though it said to be called as government policy, such decisions or government policy shall be held void by the Supreme Court.

9

AIR 1996 SC 149, 159, 163. Pleasant Stay Hotel v. Palani Hills Conservation council, 1995 (6) SCC 136, 139, 140.

10

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A STUDY ON THE BUTTERFLY DIVERSITY IN RAJAH SERFOJI GOVT. COLLEGE CAMPUS, THANJAVUR, TAMILNADU K.RAMESHKUMAR*, G.GNANAMUTHU AND R.SHANKAR Pheromone Research Lab P.G. & Research Department of Zoology Rajah Serfoji Govt. College (Autonomous) Thanjavur – 613 005, Tamilnadu, India *e-mail: [email protected]

Butterflies are the most exciting and beautiful creatures among the insect group. The butterfly diversity is high in the tropics compared to temperate regions of the world. It has important ecosystem roles including pollination and indicators of ecosystem health because they are very sensitive to microclimatic change in the habitat. Seasonal fluctuations are influenced by environmental factors like temperature, photoperiod, rainfall, humidity, availability of food and vegetation cover such as herbs and shrubs. Modified habitats often negatively influenced the butterfly species and their dynamics.The campus selected for the present study is rich in trees, flowering herbs and shrubs found to supports a large number of native as well as exotic butterfly species. Conservation of biological diversity requires detailed information on the geographic distribution of organisms. Based on the above information, the present study is aimed to know the butterfly diversity in Rajah Serfoji Government College campus. In this study, 14 different species of butterflies were identified and they belonged to four different families (Papilionidae, Nymphalidae, Pieridae, Hesperiidae). Among the families Papilionidae (5 species), Nymphalidae (4 species), Pieridae (4 species), and Hesperiidae(1 species) were identified in the campus. The species Atrophaneura hector (Papilionidae), Danacus chrysippus (Nymphalidae) and Delias eucharis (Pieridae) are dominantly present when compared to others. In addition, the present study also provides the basic information about the butterfly distribution and also to find out their occurrence, status as well as to create awareness for their conservation.

Keywords: Butterfly, Papilionidae, Nymphalidae, Pieridae, Diversity, Ecosystem

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Introduction Butterflies are the most tantalizing and beautiful creatures among the insect group. These are perhaps the most studied and well-known insect groups (Scott, 2001).There are 16,823 species recorded from all over the world among them 1501 species of butterflies are recorded from India (Gaonkar, 1996). Butterflies are taxonomically well studied group which have received a reasonable amount of attention throughout the world (Ghazoul, 2002). Butterfly community assembly and the factors which influence, it have long been a topic of interest to ecologists and conservationists. Human dominated landscape form a substantial and everincreasing amount of the earth’s land surface. Modified habitats often negatively influence butterfly species and their dynamics (Gascon, et al., 1999; Rickets, et al., 2001). Conservation of global biodiversity has become the issue of prime importance in recent decades (Turner et al., 1990; Ehrlich and Wilson, 1991). Conservationists around the globe are battling with conservation challenges under the ever accelerating threats of anthropogenic disturbances to biodiversity. Seasonality is a common phenomenon in insect populations. Seasonal fluctuations are often influenced by environmental factors including temperature, photoperiod, rainfall, humidity, variation in the availability of food resources, and vegetation cover such as herbs and shrubs (Anu 2006; Anu, et al., 2009; Shanthi, et al., 2009; Tiple and Khurad 2009). Butterflies have important ecosystem roles including pollination, and they are useful in studies of population and community ecology (Pollard, 1991) as indicators of ecosystem health because they are very sensitive to changes in microclimate and habitat (Erhardt, 1985; Kremen, 1992).Seasonal occurrences of butterfly common for only a few months and rarely absent in others. The seasons when they are rare or not active as adults are usually spent either as caterpillars or as pupae. The months when the adults are active are called the “flight period”. Distinct flight periods naturally imply seasonality of the early stages of butterflies as well (Kunte, 2000). Butterflies undergo four different life stages: egg, larva, pupa, and adult. Each life stages may require specific habitat needs, such as a particular host plant for the larva to eat or a flower nectar source for the adult to gain nutrients. Except for some migratory species, the presence of adult butterflies typically indicates that a breeding population occurs in the surrounding area.

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Moreover, adult butterflies are relatively large (compared to the other life stages), easy to identify and occur in a wide variety of different habitats (Pollard, 1991). The present investigation is aimed to study the butterflies in Rajah Serfoji Govt. College campus and also to find out their occurrence, status as well as to create awareness for their conservation. In addition, the study also providing the basic information regarding butterfly species for further studies related to biodiversity. The college campus is one of the beautiful natures of the Thanjavur, where the birds can get cover. Hence, the area should be wisely used without disturbing the activity of the butterflies and instead encouraging number of species in the area. Material and methods Thanjavur district lies as the east coast of Tamilnadu between 09°50 N to 11°25 N and 78°45 to 70°25 E. The study area, Rajah Serfoji Government College campus is located at 10°48°N 79° 09° E10.8° N 79.15° E approximately in the center of the Indian state of Tamil Nadu. The College is nearly about 110 acres located in the center of the Cauvery delta. In this campus variety of herbs, shrubs, plants and butterflies are present. The climate is typically tropical and humid. The southwest monsoon during June-September accounts for the major share of rainfall compared to the northeast monsoon of October –December. December and January are the coldest months and April – May are the hottest during which time the average daily maximum and minimum temperature are around 35°C and 20°C respectively. In the present study, the field observation was carried out during November 2012 to February 2013. A daily survey was done by systematically walked on the college campus to observe the butterflies. Butterflies were mostly observed during the most active period of the day (from 06.00 to 09.00hrs and 17.00 to 18.00hrs). Observations were carried out with the aid of binoculars and take photos.Field characteristics and the number of butterflies were counted with species wise and noted on data sheets. Results and Discussion The present study on butterflies’ community structure of the Rajah Serfoji Government College, Thanjavur revealed the presence of 14 species of butterfly belonging to 4 families. Among this Atrophaneura aristolochiae, Atrophaneura hector, Papilio demoleus, Papilio 29

dravidarum and Papilio polytes belongs to Papilionidae family,

Acraea terpsico, Danaus

chrysippus, Hypolimnas bolina and Mycalesis perseus belongs to Nymphalidae family, Cepora nerissa, Delias eucharis, Eurema hecable and Pieris rapae are belongs to Pieridae family and Pelopidas mathias blongs to Hesperiidae family (table.1).The college campus vegetative patterns are suitable for the growth and survival of the butterflies and other living things. A large number of exotic and native herbs, shrubs and plants are present in the college campus.Almost all these plants provide some cover and food for the butterflies throughout the year.The butterfly distribution are expected to cover with the distribution of their host plants even at small scales and type of vegetation may reflect difference in the composition of butterfly communities among habitats at the generic and family level (Beccaloni; 1997). Our results also confirmed the above finding, because our college vegetation and environment favourable for the growth and survival of the butterflies.India has more than 1,400 species of butterflies, 330 of them in the Western Ghats alone, and of which 37 are endemic (Kunte, 2008). The species Atrophaneura hector (Papilionidae), Danacus chrysippus (Nymphalidae) and Delias eucharis (Pieridae) are dominantly present in the college campus when compared to others Conclusion The present study concluded that in Rajah Serfoji Government College campus 14 species of butterflies belonging to 4 families. Among the species Atrophaneura hector (Papilionidae), Danacus chrysippus (Nymphalidae) and Delias eucharis (Pieridae) are dominantly present when compared to others.In addition, educating innovation of the student communities about the impacts to encourage the management of the college campus to provide suitable habitat for native butterfly species conservation. REFERENCES Anu, A., (2006).Entomofaunal Dynamics and Biochemistry of Litter Decomposition in a Natural Forest with Special Reference to the Systematics of Dung Beetles (Coleoptera: Scarabaeinae). Ph.D. Dissertation.University of Calicut, Kerala, India. Anu, A., Sabu, T.K., and Vineesh, P.J., (2009). Seasonality of litter insects and relationship with rainfall in a wet evergreen forest in south Western Ghats. J. Insect Sci. 9: 46.

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Beccaloni, G.W., (1997). Vertical stratification of the ithomiine butterfly (Nymphalidae: Ithomiinae) mimicry complexes: the relationship between adult flight height and larval hostplant height. Biol. J. Lin. Soc. 62: 313–341. Ehrilich, P.R., and Wilson, E.O., (1991). Biodiversity studies: science and policy. Science.253:750-752. Erhardt, A., (1985). Diurnal Lepidoptera: sensitive indicators of cultivated and abandoned grassland. J. Applied Eco. 22: 849-862. Gaonkar, H., (1996). Butterflies of the Western Ghats, India, including Srilanka; A biodiversity assessment of a threatened mountain system. Center for Ecological Sciences, IISc., Bangalore and the Natural History Museum, London. 51. Gascon C, Lovejoy TE, Bierregaard, RO., Malcolm ,J.R., Stouffer P.C., Vasconcelos HL, Laurance WF, Zimmerman, B., Tocher M , and Borges, S., 1999. Matrix habitat and species richness in tropical forest remnants. Biol. Conserv. 91: 223–229. Ghazoul, J., (2002). Impact of logging on the richness and diversity of forest butterflies in a tropical dry forest in Thailand. Biodivers. Conserv. 11: 521–541. Kremen, C.,(1992). Assessing the indicator properties of species assemblages for natural areas monitoring. Ecol. Applications 2: 203-217. Kunte, K.J., (2000). Butterflies of peninsular India.Indian Academy of Sciences, Bangalore and University Press, Hyderabad. Kunte, K.J., (2008). The Wildlife (Protection) Act and conservation prioritization of butterflies of the Western Ghats, South Western India. Curr. Sci. India 94: 729-735. Pollard, Ε., (1991). Monitoring butterfly numbers; in Monitoring for conservation and ecology (ed.) F Β Goldsmith (London: Chapman and Hall) p· 87 Rickets, T.H., Daily, G.C., Ehrlich, P.R., and Fay, J.P., (2001). Countryside biogeography of moths in a fragmented landscape: biodiversity in native and agricultural habitats. Conserv. Biol., 15: 378–388. Scott James, A., (2001). The butterflies of North America. A Natural History and Field Guide, Stanford University Press. Stanford. Shanthi, R., Hussain, K.J., and Sanjayan, K.P., (2009). Influence of weather on the incidence of sucking pest complex on summer-irrigated cotton crops of Tamil Nadu. Hexapoda16(1): 89-92. Tiple, A.D., and Khurad A.M., (2009). Butterfly Species Diversity, Habitats and Seasonal Distribution in and around Nagpur City, Central India. World J.Zoo.4(3): 153-162

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Table.1 List of identified butterflies in R.S.G. College Campus, Thanjavur, Tamilnadu S.No.

Name of the butterflies

Families

1

Atrophaneura aristolochiae

Papilionidae

2

Atrophaneura hector

Papilionidae

3

Papilio demoleus

Papilionidae

4

Papilio dravidarum

Papilionidae

5

Papilio polytes

Papilionidae

6

Acrae aterpsico

Nymphalidae

7

Danaus chrysippus

Nymphalidae

32

8

Hypolimnas bolina

Nymphalidae

9

Mycalesis perseus

Nymphalidae

10

Cepora nerissa

Pieridae

11

Deliase ucharis

Pieridae

12

Eurema hecable

Pieridae

13

Pieris rapae

Pieridae

14

Pelopidas mathias

Hesperiidae

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BIOPIRACY AND COMMERCIALIAZATION OF NEW PRODUCTS V.Murugesan 1, M.Sharmila 2 & K.Jeyaprakash 3 1. Asst Professor, Dept of Statistics, Rajah Serfoji Govt.College (A), Thanjavur -05 2. Asst Professor, Dept of Chemistry, Rajah Serfoji Govt.College (A), Thanjavur -05 3. Head, Dept of Biochemistry, Rajah Serfoji Govt.College (A), Thanjavur -05

INTRODUCTION Bioprospecting is the process of discovery and commercialization of new products based on biological resources. Despite being intuitively helpful, bioprospecting has only recently begun to incorporate indigenous knowledge in focusing screening efforts for bioactive compounds. [1] Bioprospecting also includes biopiracy, the exploitative appropriation of indigenous forms of knowledge by commercial actors, and bioprospecting includes the search for previously unknown compounds in organisms that have never been used in traditional medicine before.[2] Biopiracy is a situation where indigenous knowledge of nature, originating with indigenous peoples, is used by others for profit, without permission from and with little or no compensation or recognition to the indigenous people themselves.[3] For example when bioprospectors draw on indigenous knowledge of medicinal plants which is later patented by medical companies without recognizing the fact that the knowledge is not new, or invented by the patenter, and depriving the indigenous community to the rights to commercial exploitation of the technology that they themselves had developed. Critics of this practice such as Greenpeace,[4] claim these practices contribute to inequality between developing countries rich inbiodiversity, and developed countries hosting companies that engage in 'biopiracy'. FAMOUS CASES The Maya ICBG bioprospecting controversy took place in 1999-2000, when the International Cooperative Biodiversity Groupled by Ethnobiologist Dr. Brent Berlin was accused of being engaged in unethical forms of bioprospecting by several NGOsand indigenous organizations. The ICBG aimed to document the biodiversity of Chiapas, Mexico and the ethnobotanical knowledge of the indigenous Maya people - in order to ascertain whether there were possibilities of developing medical products based on any of the plants used by the indigenous groups.[5][6] The Maya ICBG case was among the first to draw attention to the problems of distinguishing between benign forms of bioprospecting and unethical biopiracy, and to the difficulties of securing community participation and prior informed consent for would-be bioprospectors.[7] 34

THE ROSY PERIWINKLE The rosy periwinkle case dates from the 1950s. The rosy periwinkle, while native to Madagascar, had been widely introduced into other tropical countries around the world well before the discovery of vincristine. This meant that researchers could obtain local knowledge from one country and plant samples from another. The use of the plant as a cure for diabeteswas the original stimulus for research. Effectiveness in the treatment of both Hodkin's Disease and leukemia were discovered instead.[8] Different countries are reported as having acquired different beliefs about the medical properties of the plant.[9] The Hodgkin's lymphoma chemotherapeutic drug vinblastine is derivable from the rosy periwinkle.[10]

THE NEEM TREE In 1994, the U.S. Department of Agriculture and WR Grace received a European patent on methods of controlling fungal infections in plants using a composition that included extracts from the neem tree (Azadirachta indica), which grows throughoutIndia and Nepal.[11][12][13] In 2000 the patent was successfully opposed by several groups from EU and India including the EU Green Party, Vandana Shiva, and theInternational Federation of Organic Agriculture Movements (IFOAM) on the basis that the fungicidal activity of neem extract had long been known in Indian traditional medicine.[13] WR Grace appealed, and lost that appeal in 2005.[14]

THE ENOLA BEAN The Enola bean is a variety of Mexican yellow bean, so called after the wife of the man who patented it in 1999.[15] The allegedly distinguishing feature of the variety is seeds of a 35

specific shade of yellow. The patent-holder subsequently sued a large number of importers of Mexican yellow beans with the following result: "...export sales immediately dropped over 90% among importers that had been selling these beans for years, causing economic damage to more than 22,000 farmers in northern Mexico who depended on sales of this bean."[16] A lawsuit was filed on behalf of the farmers, and on April 14, 2005 the US-PTO ruled in favor of the farmers. An appeal was heard on 16 January 2008, and the patent was revoked in May 2008. An appeal to the court against the revocation was unsuccessful (Decided October 2009).[17]

BASMATI RICE In 2000, the US corporation RiceTec (a subsidiary of RiceTec AG of Liechtenstein) attempted to patent certain hybrids ofbasmati rice and semidwarf long-grain rice.[18] The Indian government intervened and several claims of the patent were invalidated. Meanwhile, the European Commission has agreed to protect basmati rice under its regulationspertaining to geographical indications. HOODIA Hoodia, a succulent plant, originates from the Kalahari Desert of South Africa. For generations it has been known to the traditionally living San people as an appetite suppressant. In 1996 South Africa’s Council for Scientific and Industrial Researchbegan working with companies, including Unilever, to develop dietary supplements based on hoodia.[19][20][21][22] Originally the San people were not planned to receive any benefits from the commercialization of their traditional knowledge, but in 2003 the South African San Council made an agreement with CSIR in which they would receive from 6 to 8% of the revenue from the sale of Hoodia products.[23] In 2008 after having invested €20 million in R&D on hoodia as a potential ingredient in dietary supplements for weight loss, Unilever terminated the project because their clinical studies did not show that hoodia was safe and effective enough to bring to market.[24]

36

PATENT LAW One common misunderstanding is that pharmaceutical companies patent the plants they collect. While obtaining a patent on a naturally occurring organism as previously known or used is not possible, patents may be taken out on specific chemicals isolated or developed from plants. Often these patents are obtained with a stated and researched use of those chemicals. Generally the existence, structure and synthesis of those compounds is not a part of the indigenous medical knowledge that led researchers to analyze the plant in the first place. As a result, even if the indigenous medical knowledge is taken as prior art, that knowledge does not by itself make the active chemical compound "obvious," which is the standard applied under patent law. In the United States, patent law can be used to protect "isolated and purified" compounds - even, in one instance, a new chemical element (see USP 3,156,523). In 1873, Louis Pasteur patented a "yeast" which was "free from disease" (patent #141072). Patents covering biological inventions have been treated similarly. In the 1980 case of Diamond v. Chakrabarty, the Supreme Court upheld a patent on a bacterium that had been genetically modified to consume petroleum, reasoning that U.S. law permits patents on "anything under the sun that is made by man." The United States Patent and Trademark Office (USPTO) has observed that "a patent on a gene covers the isolated and purified gene but does not cover the gene as it occurs in nature".[26] Also possible under US law is patenting a cultivar, a new variety of an existing organism. The patent on the enola bean (now revoked) was an example of this sort of patent. The intellectual property laws of the US also recognize plant breeders' rights under the Plant Variety Protection Act, 7 U.S.C. §§ 2321-2582.[27] Convention on Biological Diversity (CBD) The CBD came into force in 1993. It secured rights to control access to genetic resources for the countries in which those resources are located. One objective of the CBD is to enable lesser-developed countries to better benefit from their resources and traditional knowledge. 37

Under the rules of the CBD, bioprospectors are required to obtain informed consent to access such resources, and must share any benefits with the biodiversity-rich country.[28] However, some critics believe that the CBD has failed to establish appropriate regulations to prevent biopiracy. Others claim that the main problem is the failure of national governments to pass appropriate laws implementing the provisions of the CBD.[29] The Nagoya Protocol to the CBD (negotiated in 2010, expected to come into force in 2014) will provide further regulations. The CBD has been ratified by all countries in the world except for Andorra, Holy See and United States. The 1994 Agreement on Trade-Related Aspects of Intellectual Property Rights (TRIPs) and the 2001 International Treaty on Plant Genetic Resources for Food and Agriculture are further relevant international agreements. Bioprospecting contracts The ethical debate has sparked a new branch of international patent and trade law. Bioprospecting contracts lay down the rules, between researchers and countries, of benefit sharing and can bring royalties to lesser-developed countries. However, the fairness of these contracts has been a subject of debate. Unethical bioprospecting contracts (as distinct from ethical ones) can be viewed as a new form of biopiracy. An extensively discussed example of a bioprospecting contract is the agreement between Merck and INBio of Costa Rica.[30] On June 14, 2011, Colombia approved a policy for the sustainable commercial use of its biodiversity resources, primarily through the development of biotechnology research. It includes plans to set up a national company for bioprospecting to link up with the commercial sector and will be backed with US$14 million in government funds over the next four years.[31][32] TRADITIONAL KNOWLEDGE OF DATABASES In response to concerns of biopiracy raised by research into turmeric, neem and basmati rice, the Government of India has been translating and publishing ancient manuscripts containing old remedies in electronic form, and in 2001 the Traditional Knowledge Digital Library was set up as a repository of 1200 formulations of various systems of Indian medicine, such asAyurveda, Unani and Siddha.[25][33] The texts are being recorded from Sanskrit, Urdu, Persian and Arabic; made available to patent offices in English, German, French, Japanese and Spanish. The aim is to protect India's heritage from being exploited by foreign companies. Hundreds of yoga poses are also kept in the collection. The project has been criticized by a spokesman for the pharmaceutical industry as "a solution in search of a problem". The library has also signed agreements with leading international patent offices such as European Patent Office (EPO), United Kingdom Trademark & Patent Office(UKPTO) and the United States Patent and Trademark Office to protect traditional knowledge from biopiracy as it allowspatent examiners at International Patent Offices to access TKDL databases for patent search and examination purposes.[25][34][35] The database is not available to the general public, but 38

the articles to which it refers are (though typically in a not widely understood language such as Sanskrit). In this way the database prevents subsequent patenting without making the information to which it refers readily available for public use.

REFERENCES 1. C. Haris Saslis-Lagoudakisa , Vincent Savolainenb,c, Elizabeth M. Williamson, (2012) Phylogenies reveal predictive power of traditional medicine in bioprospecting, PNAS, Vol 109, 15385-15390. 2. Cluis, Corinne (2013). "Bioprospecting: A New Western Blockbuster, After the Gold Rush, the Gene Rush". The Science Creative Quarterly (8) (The Science Creative Quarterly (University of British Columbia)). Retrieved2013-11-04. 3. "biopiracy - definition of biopiracy by the Free Online Dictionary, Thesaurus and Encyclopedia". Thefreedictionary.com. Retrieved 2013-11-03. 4. "Agriculture and Food". Green Peace Australia Pacific: What We Do: Food. Greenpeace. Retrieved 2013-11-04. 5. Cori Hayden (2003). When Nature Goes Public: The Making and Unmaking of Bioprospecting in Mexico. Princeton University Press. pp. 100–105. ISBN 978-0-69109556-1. Retrieved 2013-11-04. 6. Feinholz-Klip, Dafna; García Barrios, Luis; Cook Lucas, Julie (2009). "The Limitations of Good Intent: Problems of Representation and Informed Consent in the Maya ICBG Project in Chiapas, Mexico". In Wynberg, Rachel, Doris Schroeder & Roger Chennells (eds.). Indigenous Peoples, Consent and Benefit Sharing. Springer Netherlands. pp. 315– 331. doi:10.1007/978-90-481-3123-5_17.ISBN 978-90-481-3123-5. 7. James V. Lavery (2007). "Case 1: Community Involvement in Biodiversity Prospecting in Mexico". Ethical Issues in International Biomedical Research: A Casebook. Oxford University Press. pp. 21–43.ISBN 978-0-19-517922-4. Retrieved 2013-11-04. 8. http://muse.jhu.edu/journals/jaf/summary/v117/117.465hafstein.html 9. "A traditional brew leads to cancer cure". Smithsonian Institution: Migrations in history: Medical Technology. Smithsonian Institution. Retrieved 2013-11-04. 10. Karasov, C. (2001). "Who Reaps the Benefits of Biodiversity?". Environmental Health Perspectives(Environmental Health Perspectives, Vol. 109, No. 12) 109(12): A582– A587. doi:10.2307/3454734.JSTOR 3454734. PMC 1240518. PMID 11748021. 11. European Patent EP0436257B1 12. Karen Hoggan for the BBC. May 11, 2000 Neem tree patent revoked 13. Cormac Sherida. Nature Biotechnology 23(5):511-12. May 2005. EPO neem patent revocation revives biopiracy debate 14. BBC News, March 9, 2005 India wins landmark patent battle

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15. L. Pallottinia, E. Garciab, J. Kamic, G. Barcacciaa and P. Gepts (1 May 2004). "The Genetic Anatomy of a Patented Yellow Bean". Crop Science 44 (3): 968– 977.doi:10.2135/cropsci2004.0968. 16. Goldberg, Danielle (2003). "Jack and the Enola Bean".TED Case Studies Number xxx. Danielle Goldberg. Retrieved 2013-11-04. 17. Neil (2009-07-22). "New legal decision against Enola bean". CIAT: News: Ecoefficiency in Action: Crops @en: Beans @en. International Center for Tropical Agriculture (CIAT). Retrieved 2013-11-04. 18. U.S. Patent No. 5,663,484 19. Maharaj, VJ, Senabe, JV, and Horak, RM. 2008. Hoodia, a case study at CSIR. Science real and relevant: 2nd CSIR Biennial Conference, CSIR International Convention Centre Pretoria, 17&18 November 2008, pp 4 20. Rachel Wynberg; Doris Schroeder; Roger Chennells (30 September 2009). Indigenous Peoples, Consent and Benefit Sharing: Lessons from the San-Hoodia Case. Springer. ISBN 978-90-481-3123-5. Retrieved 2013-11-04. 21. Vermeylen, Saskia (2007). "Contextualizing 'Fair' and 'Equitable': The San's Reflections on the Hoodia Benefit-Sharing Agreement". Local Environment: The International Journal of Justice and Sustainability (Informa UK Limited) 12 (4): 423– 436.doi:10.1080/13549830701495252. Retrieved 2013-11-04. 22. Wynberg, Rachel (2013-10-13). "Hot air over Hoodia".Grain: Publications: Seedling. Grain. Retrieved 2013-11-03. 23. Foster, Laura A. (April 2001). "Inventing Hoodia: Vulnerabilities and Epistemic Citizenship in South Africa". UCLA Center for the Study of Women: CSW update (UCLA Center for the Study of Women). Retrieved2013-11-04. 24. http://www.unilever.com.au/sustainability/nutrition/ 25. "Know Instances of Patenting on the UES of Medicinal Plants in India". PIB, Ministry of Environment and Forests. May 6, 2010. 26. "Department of Commerce: United States Patent and Trademark Office [Docket No. 991027289-0263-02] RIN",Federal Register: Notices (Office of the Federal Register of the National Archives and Records Administration) 66 (4), 2001-01-05: 1092–1099, retrieved 2013-11-04 27. Chen, James Ming (2006). "The Parable of the Seeds: Interpreting the Plant Variety Protection Act in Furtherance of Innovation Policy". Notre Dame Law Review (Social Science Electronic Publishing, Inc.) 81 (4): 105–166. Retrieved 2013-11-04. 28. CBD stating that the benefits arising from the use of genetic resources should be shared in a fair and equitable way (Rau, 2010)/ 29. "Policy Commissions". International Chamber of Commerce: About ICC. International Chamber of Commerce. Retrieved 2013-11-03. 30. Eberlee, John (2000-01-21). "Assessing the Benefits of Bioprospecting in Latin America". IDRC Reports Online. IDRC. Retrieved 2013-11-03. 31. Fog, Lisbeth (2011-06-07). "Columbia to commercialise its biodiversity". Sci Dev Net: Global: Biodiversity: News. SciDev.Net. Retrieved 2013-11-04. 40

32. "Política Para El Desarrolo Comercial De La Biotecnología A Partir Del Uso Sostenible De La Biodiversidad", Documento Conpes (in Spanish) (Bogotá: Consejo Nacional de Politica Económica y Social, República de Columbia, Departamento Nacional de Planeación) 3697, 2011-06-14, retrieved 2013-11-04 33. "Traditional Knowledge Digital Library". Council of Scientific and Industrial Research and Department of Ayurveda, Yoga & Naturopathy, Unani, Siddha and Homeopathy. Retrieved 2013-11-03. 34. Koshy, Jacob P.; gen patent claim (2010-04-28). "CSIR wing objects to Avesth". live mint & The Wall Street Journal (HT Media). Retrieved 2013-11-04. | 35. "India Partners with US and UK to Protect Its Traditional Knowledge and Prevent BioPiracy" (Press release). Press Information Bureau, Ministry of Health and Family Welfare, Government of India. 2010-04-28. Retrieved2013-11-04.

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A View of Soil Fungal Flora in Thiruvarur District, East Coast of Tamil Nadu, India Arulmozhi,R.* and Kannahi,M PG and Research Department of Microbiology, Sengamala Thayaar Educational Trust Women’s College, Sundarakkottai, Mannargudi-614001, Tamil Nadu, India. Email id: [email protected] Abstract Soil represents a favorable habitat for microorganisms and inhabited by a wide range of microorganisms. Microorganisms are found in large numbers in soil, usually one to ten million microorganisms is present per gram of soil with a dominant number of fungi. The contribution of soil fungi was very significant in many soil functions such as supporting the growth of plants, absorbing, neutralizing and transforming compounds that might otherwise become pollutants in the environment. This study deals with fungal diversity at various sites of traditional paddy field in Thiruvarur district. The soil from paddy field of Sundarakkottai, Laxmangudi, Vellore, Mannargudi, Manali and Serankulam was infected by the fungi Tricoderma, Mucor, Fusarium, Arthrobotrys, Rhizopus, Rhizoctonia and Aspergillus. In the present study, it was concluded that mostly Mucor is dominated followed by Trichoderma and Rhizopus. Keywords: Habitat, Diversity, Fungi, Microorganisms and Soil

INTRODUCTION Soil is a complex system. Many biological processes take place in soil and determine functions that provide various services within ecosystes: turn-over of organic matter, symbiotic and non-symbiotic atmospheric nitrogen fixation, denitrification, aggregation etc., Filters and remediates anthropogenic pollutants, and enables food production (Kennedy and Smith, 1995; Richards, 1987).

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One particularly significant component of soil is the microorganisms. Soil is a medium with solids, liquids and gasses in which the mineral and organic particles form differently sized aggregates that delimit pores. This organization creates micro-environments that are suited to microbial activity hot spots (Gillard et al., 2003). Microorganisms are beneficial in increasing the soil fertility and plant growth as they are involved in several biochemical transformation and mineralition activities in soil. Type of cultivation and crop management practices found to have greater influence on the activity of soil microflora (Mc Gill et al., 1980). Continuous use of chemical fertilizers over a long period may cause imbalance in soil microflora and therby indirectly affect biological properties of soil leading to soil degradation (Manickam et al., 1972).

Fungi are fundamental for soil ecosystem functioning (Warcup, 1950). Especially in forest and agricultural soils; they play a key role in much essential process such as organic matter decomposition and elemental release by mineralization (Christensen et al., 1989). The rate of biodegradation depends on environmental factors, numbers and types of microorganisms present and the enzymatic process leading to the disappearance of the parent molecular structure and the formation of smaller organic species. Some of which are directly usable for cell anabolism and are converted to CO2 and H2O ultimately (Mishra et al., 1991)

Material and methods: About six soil samples were collected from the Thiruvarur district .i.e., Sundarakkottai, Laxmangudi, Vellore, Mannargudi, Manali and Serankulam. The potato dextrose agar media was selected to grow fungi of soils. Results and discussion Fungal cultures were isolated and prepared temporary slide and observed under microscope sketches were drawn and photography was done. In the present investigations seven genus of fungi were isolated from the soil of paddy field. Tricoderma and Mucor is observed. In Laxmangudi soil Fusarium and Arthrobotrys fungus are observed. In the soil of Manali Rhizopus and Mucor are observed. From Mannargudi soil Tricoderma, Rhizoctonia and Mucor are observed and From Sundarakkottai soil Mucor and Aspergillus are observed. In the Serankulam soil Rhizopus and Mucor are observed.

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Comparative study was carried out among paddy field soil of different talukas it was found that Mucor is dominated followed by Tricoderma and Rhizopus. Selvaraj kalaiselvi and Annamalai Panneerselvam (2011) worked on ecology of soil fungi in paddy field of Tamilnadu, they isolated various fungal species in which the dominated species was Aspergillus nigre but in present study Mucor is dominent . On the other hand V. Manimegalai et al worked on paddy field of Thanjavur district, Tamilnadu, where dominant species are Aspergillus niger and Aspergillus flavus. Sethilkumar et al. (2009) worked on soil samples of three different places along the Muthupet mangroves in Tamilnadu out of 22 species Aspergillus and Penicillium were represented as dominated species. J.M Benila Smily et al. (2012) found the fungal species in rice field with paddy alone showed 13 species and Aspergillus is dominated. Kadar et al. (1999) found Aspergillus genus is dominated in their work Microscopic slide observation of soil fungi

Tricoderma

Aspergillus

Mucor

Rhizopus

Fusarium

Arthrobotrys

CONCLUSION Generally, top soil contains high organic matter, which in the presence of adequate moisture supply is acted upon by the microorganisms to decompose the complex organic residues into simpler forms; hence, microbial counts are generally higher in the surface soil layer 44

as compared to the lower depths. Soil of paddy field Sundarakkottai, Laxmangudi, Vellore, Mannargudi, Manali and Serankulam of was infected by the Tricoderma, Mucor, Fusarium, Arthrobotrys, Rhizopus, Rhizoctonia and Aspergillus. After comparative study, it was concluded that mostly Mucor is dominated followed by Tricoderma and Rhizopus.

The important factors influencing the variation in the population of fungi in the present study could possibly due to temperature, organic nitrogen and moisture content of the soil. The paddy field soil was subjected to disturbances such as irrigation, fertilizer and agricultural practices resulting in more homogeneity of soil which did not allow relatively wide fluctuation in the population of fungi. Agriculture would not be possible without microorganisms. Therefore, the soil depends upon the microorganisms for the fertility. REFERENCES

1. Benila Smily JM, Vinoy Jacob and Ravi Kumar M (2012) Soil microflora of paddy fields among different rice forming system. J. Acad. Indu. Res., 1 (1): 50-52. 2. Christensen M (1989) A view of fungal ecology, Mycologia, 81: 1-19. 3. Gaillard V, Chenu C and Recous S (2003) Carbon mineralisation in soil adjacent to plant residues of contrasting biochemical quality. Soil biology and biochemistry, 35: 93-99. 4. Kadar AJ, Omar O and Fing LS (1999) Isolation of cellulolytic fungi from the Balio Highlands, Sarawork. ASEAN review of Biodiversity and Enviornmental conservation, 1:1-3. Kennedy AC and Smith KL (1995) Soil microbial diversity and the sustainability of agricultural soils, Plant and soil, 170:75-86. 5. Manickam TS and Venkataraman CR (1972) Influence of Fertilization and different tillage systems on soil microflora, Madras Agriculrtural Journal, 59: 508-512. 6. Manimegalai V, Ambikapathy V and Panneerselvam A (2011) Population dynamics of soil mycoflora in the paddy field of Thanjavur district, Tamilnadu. European Journal of expermintal Biology, 2011, 1 (3) : 14-19. Mc. Gill WB, Cannon KR, Robertson JA and Coock GD (1980) Dynamics of soil microbial biomass and water stable organic carbon in Breton. L after fifty years of cropping to two rotations, Canadian journal of soil science, 66: 1-19. 45

7. Mishra PC, Manorama Patri and Madumita Panda (1991) Growth of water hyacinth and its efficiency in the removal of pollution load from industrial waste, Journal of Ecotoxicology and Environmental Monitoring, 1(13):218-224. 8. Richards BN (1987) Mineral cycling processes. In: The microbiology of terrestrial ecosystems. . John Wiley and Sons, New York, pp 177-221. 9. Selvaraj Kalaiselvi and Annamalai Panneerselvam (2011) Ecology of soil fungi in paddy field of Tamilnadu-Thanjavur district. Peligia Research Library, Der chemicasinica, 2 (2) : 9-19. 10. Sethilkumar G. Madharaj P. Kanimoshi K and Panneerselvam A (2009) J. Microb. World, 11 (1): 31-36. 11. Warcup JH (1950) The Soil-plate method for isolation of fungi from Soil, Nature, Lond, 117166.

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EFFECT OF PSEUDOMONAS FLUORESCENS ON BIOCHEMICAL CHANGES OF SAMANEA SAMAN UNDER NURSERY CONDITION E. Mohan1 M.Kannan2 and K.Rajendran1 1- Research Department of Botany, Thiagarajar College, Madurai -9, Tamil Nadu 2- Department of Botany, NMSS.Vellaichamy Nadar College, Nagamalai, Madurai Email: [email protected] ABSTRACT A Nursery experiment was conducted to evaluate the effectiveness of Pseudomonas fluorescens on biochemical changes of Samanea saman. The plants treated with bioinoculant (Pseudomonas fluorescens) had increase the biochemical contents such as chlorophyll a, chlorophyll b, total chlorophyll, carotenoid, soluble protein, soluble sugar content 180 days after inoculation when compared with the control plants. The results revealed that this treatment could be enhancing the biochemical parameters in Samanea saman seedlings. Key words: Pseudomonas fluorescens, Samanea saman, Biochemical content INTRODUCTION Samanea saman belongs to the second largest family of flowering plants, Leguminoseae. It is easily recognized by its characteristic umbrella-shaped canopy, reaching a height of 20 to 25 meters, with the branches wide spreading. The tiny flowers are massed in pinkish heads 5–6 cm across and about 4 cm in height. Pods are fleshy, straight, more or less flat and black when ripe. Seeds 16 to 20, rounded, truncate at one end, pointed at the other, dark reddish brown with a paler ring at each side. The plant is used traditionally in diarrhea, intestinal diseases, stomach ache, colds and headache (Prasad et al., 2008). It is well known that a considerable number of bacterial species, mostly those associated with the plant rhizosphere, are able to exert a beneficial effect upon plant growth. This group of bacteria has been termed ‘plant growth promoting rhizobacteria’. Strains from genera such as Pseudomonas, Azospirillum, Burkholderia, Bacillus, Enterobacter, Rhizobium, Erwinia, Serratia, Alcaligenes, Arthrobacter, Acinetobacter and Flavobacterium are act as PGPR. Stimulation of different crops by PGPR has been demonstrated in both laboratory and field trials. Strains of Pseudomonas fluorescens have increased root and shoot elongation in various crops. (Kloepper and Schroth, 1978). The role of biofertilizers has already been proved extensively in annual crops, but its exploitation in forest species is scanty in India. In forestry, few research reports are available to demonstrate that biofertilizers stimulate the growth (Sudhir et al. 1994), biomass (Sekar et al., 1995), biochemical parameters (Niranjan et al. 1990) and uptake of nutrients (Mathur and Vyas 1996). Ultimately, the survival rate of planted seedling is enhanced. Hence, a study on the effect of biofertilizers on biochemical parameters of Samanea saman was initiated.

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MATERIALS AND METHODS Study site The study area selected was in Madurai, Tamil Nadu located at100 05’ N latitude and 780 16’ E longitude. The elevation of the study area ranges from 100-132 M above mean sea level. The temperature ranges from 210 C during winter and about 390 C during summer. The study area of Madurai received the annual rain fall of about 750 and 850 mm. The climate is subhumid to semi-arid. Seed collection Seeds of Samanea saman was collected from the single tree in Alagar hills located in Madurai district, Tamil Nadu The collected seeds were surface sterilized with 0.01 percent mercuric chloride for 30 seconds rinsed several times with sterile distilled water and dried aseptically for further germination purpose. Isolation of Pseudomonas Pseudomonas fluorescens was isolated from soil used for planting stock production. The soil particles loosely adhering were gently teased out and mixed well. The soil thus obtained was crushed in a sterile mortar and pestle and shaken with 100 ml of sterile distilled water for 10-20 min, to obtain standard soil suspension. Isolation of Pseudomonas fluorescens was made by following the serial dilutions and pour plate method using the specific King’s B medium. Mass multiplication of Pseudomonas King’s B medium was a selective one (King’s et al., 1954) was used for the isolation of Pseudomonas fluorescens. One ml of soil suspension from aliquot dilutions (10-5 to 10-8) was aseptically added to sterile petriplates containing 20 ml of sterile medium and incubated at 28±20C for 48h. After incubation, well separated individual colonies with yellow green and blue white pigments were marked and detected by viewing under UV light. The individual colonies were mass multiplied in nutrient broth and picked up with sterile loop and transferred to fresh King’s B slants and the pure cultures so obtained were stored in refrigerator at 40C for future use. Potting media and treatments Seeds were germinated in sterile potting media and transplanted to polythene bags15 days after germination in 13 x 26 cm sized, with a mixture of unsterilized Red soil: Sand: Farm yard manure (1: 2: 1), watering was done every day with the help of hand sprayer. After germination healthy uniform size seedlings were selected for experimental work and following bioinoculants (Treatments) were added in polythene bags. All the plants were kept under identical nursery condition up to 180 days. After six months the plant leaves were collected for biochemical analysis. Biochemical Determination Estimation of chlorophyll The concentration of chlorophyll was calculated using the formula of Arnon (1949). Estimation of carotenoid

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The content of total carotenoid in the 80% acetone extract was assessed using an absorbance co-efficient A473 1% =2500 according to Goodwin (1954) Estimation of Protein The protein content was estimated by the method of Lowry et al. (1951) Estimation of soluble sugar The soluble sugar can be estimated by the method of Dubois et al. (1956). RESULTS AND DISCUSSION The seedlings inoculated with Pseudomonas fluorescens has a significant enhancement in biochemical parameters like chlorophyll a, b and total chlorophyll content, carotenoid, soluble protein and soluble protein with that of control plants. A change in biochemical parameters were observed in bioinoculant treated seedlings of S. saman. Pseudomonas treated plants had shown an increased chlorophyll, carotenoid, soluble protein and soluble sugar content. This may be due to the inoculation of Pseudomonas. Biofertilizer have shown greater absorption of Mg, which may induce on chlorophyll synthesis (Jentschke et al., 2000). Similarly, Manoharan et al. (2008) reported higher chlorophyll a, b, total chlorophyll and carotenoid content in Cassia siamea, Delonix regia, Erythrinia variegata, Samanea saman and Sterculia foetida inoculated with biofertilizer. Aggarwal et al. (2012) also observed that chlorophyll content of Linum usitatissimum was improved by the application of Pseudomonas fluorescens and AM fungi along with recommended super phosphate fertilizer. The present study soluble protein and sugar content in the leaves of S. saman treated with bioinoculants was increased. It was corroborated with other findings in Moringa oleifera seedlings inoculated with A. brasilense (Kasthuri Rengamani et al., 2006), in Pongamia pinnata seedlings inoculated with Rhizobium, phosphobacteria and VAM (Vanangamudi et al., 1998) and in Ziziphus mauritiana when inoculated with AM fungi (Mathur and Vyas, 1996). Mathur and Vyas (2000) reported an enhanced synthesis of soluble sugars in Ziziphus mauritiana seedlings inoculated with AM fungi. CONCLUSION This study ascertains that Pseudomonas fluorescens could be used as growth promoting bioinoculant for enhancing the biochemical contents and quality seedling production of S. saman. It was a suitable bioinoculant for sustainable production of quality seedlings in forest nurseries of semi arid zone. Also it was more effective, eco-friendly and economically viable method for sustainable production of quality seedlings and high survival rate in the field conditions. REFERENCES Aggarwal, A., Neetu, N., Tanwar, A. and Alpa, A. 2012. Influence of Arbuscular Mycorrhizal fungi and Pseudomonas fluorescens at different superphosphate levels on linseed (linum usitatissimum L.) growth response. J. Agri. Res. 72(2): 237-243.

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Arnon, D.I. 1949. Copper enzymes in isolated chloroplast. I. Polypenol oxidase in Beta vulgaris. Plant Physiol. 24: 1-15. Dubois, M., Gilles, K.N., Hamilton, J.K., Roders, P.A. and Smith, S. 1956. Colorimetric method for determination of sugars and related substances. Anal Chem. 28: 350-356. Goodwin, T.W. 1954. Carotenoids. In: Hand book of plant Analysis. (eds.) Peach K, Tracey MW. Springer Verlag, Berlin, Vol III. pp.272-311, Jentschke, G. Brandes, B. Kuhn, A.J. Schroder, W.H. Becker, J.S and Godbold, D.L. 2000. The mycorrhizal fungus Paxillus involutus transport magnesium to Norway spruce seedings. Evidence from stable isotope labeling. Plant Soil. 220: 243-246. Kasthuri Rengamani, S, Jothibasu, M and Rajendran, K. 2006. Effect of bioinoculants on quality seedlings production of Drumstick (Moringa oleifera L.). J. Non-Timber For. Pro. 13(1): 41-46. King, E.O. Ward, M.K and Raney, D.E. 1954. Two simple media for the demonstration of Pyocianin and fluorescein. J. Lab. Clin. Med. 44: 301-307. Kloepper J.W, Schroth M.N. 1978. Plant growth-promoting rhizobacteria on radishes. In: Station de Pathologie vegetale et Phyto-bacteriologie, editor. Proceedings of the 4th International Conference on Plant Pathogenic Bacteria Vol II. Tours: Gilbert-Clary, pp. 879–82. Lowry, D. H. Rosenrough, H.J. Lawstarr, A. and Randal, R.J. 1951. Protein measurement with the Folin-Phenol reagent. J. Bio. Chem. 93: 265-275. Manoharan, P. T. Pandi, M. Shanmugaiah, V. Gomathinayagam, S and Balasubramanian, N. 2008. Effect of vesicular arbuscular mycorrhizal fungus on the physiological and biochemical changes of five different tree seedlings grown under nursery conditions. Afr. J. Biotechnol. 7 (19): 3431-3436. Mathur N and Vyas A 1996 Physiological changes in Ziziphus mauritiana by different VAM fungi. Indian For. 122(6): 501-505. Mathur, N and Vyas, A. 2000. Influence of arbuscular mycorrhizae on biomass production, nutrient uptake and physiological changes in Ziziphus mauritiana Lam. under water stress. J. Arid Environ. 45: 191-195. Niranjan R, Banwarilal S and Rao M 1990 Studies on the effect of Rhizobium and endomycorrhizal interaction in Dalbergia sissoo. In: Proceedings of a National Conference on Mycorrhiza. Hisar Agricultural University, Hisar, India. Feb. 14- 16,1990. pp.205-207. Prasad R.N, Viswanathan S, Devi JR, Nayak V, Swetha VC, Archana BR, Parathasarathy N, Rajkumar J. (2008) Preliminary phytochemical screening and antimicrobial activity of Samanea saman. J. Med. Plants Res. 2(10): 268-270. raising seedlings of fast growing trees. Indian J. For. 17(3): 243-248. 50

Sekar I, Vanangamudi K and Suresh K.K 1995 Effect of biofertilizers on the seedling biomass, VAMcolonization, enzyme activity and phosphorus uptake in the Shola tree species. Myforest 31 (4): 21-26.

Sudhir, M. U. Sonali, N.J., Paude, S.S, Gaikwad, S.J and Jawarkar, A.S. 1994. Microbial technology for raising seedlings of fast growing trees. Ind. J. For. 17(3): 243-248. Vanangamudi, K. Venkatech, A. V. Mallika, V. Ravichandran and Rai, R.S.V. 1998. Impact of biofertilizers on morphological attributes in pungam (Pongamia pinnata (L.) Pierre) seedlings. Trop. Agric. Res. Exten. 1: 7-11. ********************************************

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THE EFFECT OF IPR ON ANIMAL GENETIC RESOURCES, GENETIC DIVERSITY, GENETIC IMPROVEMENT AND THE LIVELIHOODS OF LIVESTOCK KEEPERS Selvam, R.*, Panneerselvam, S., Thiruvenkadan, A.K., Ponnudurai, G., Ramesh Saravanakumar, V, and Thilak Pon Jawahar, K *Assistant Professor, Department of Animal Genetics and Breeding, Veterinary College and Research Institute, Tirunelveli – 627 001

Intellectual property refers to creations of the mind: inventions, literary and artistic works and symbols, names and images used in commerce. Intellectual property is divided into two categories: Industrial Property includes patents for inventions, trademarks, industrial designs and geographical indications. Copyright covers literary works (such as novels, poems and plays), films, music, artistic works (e.g., drawings, paintings, photographs and sculptures) and architectural design. Rights related to copyright include those of performing artists in their performances, producers of phonograms in their recordings, and broadcasters in their radio and television programs (WIPO, 1970).

The IP rights are outlined in Article 27 of the Universal Declaration of Human Rights, which provides for the right to benefit from the protection of moral and material interests resulting from authorship of scientific, literary or artistic productions. The importance of intellectual property was first recognized in the Paris Convention for the Protection of Industrial Property (1883) and the Berne Convention for the Protection of Literary and Artistic Works (1886). Both treaties are administered by the World Intellectual Property Organization (WIPO, 1970).

IPR are largely territorial in nature except copyright, which is global in nature. Under the Paris Convention Treaty (March 20, 1883, as revised at Stockholm on July 14, 1967, and as amended on September 28, 1979) a group of countries agreed to work with each other to allow patent applicants in these countries a one-year period in which to file an application in one of the other countries without losing the benefit of their filing date.

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The reasons for promote and protect intellectual property are

  

 IPRs give creators exclusive rights to their creations, thereby providing an incentive for the author or inventor to develop and share the information rather than keep it secret.   The legal protection of new creations encourages the commitment of additional resources for further innovation.   The promotion and protection of intellectual property encourages economic growth, creates new jobs and industries, and enhances the quality and enjoyment of life.   An efficient intellectual property system can help all countries to realize intellectual property’s potential as a catalyst for economic development and social and cultural well-



being.   The intellectual property system helps strike a balance between the interests of innovators and the public interest, providing an environment in which creativity and invention can flourish, for the benefit of all. 

IPR rewards the creativity and it encourages economic growth. Diamond vs. Chakrabarty law case open the ways for patenting of higher forms of life. First animal patent is for “Onco mouse” by Harvard University (1984). (Lessor, 2002 and Rothschild, 2002). Patent Patent is exclusive right given to the inventor for his invention, it represents a legal monopoly granted by the respective country's government to an inventor (specific to a geographic region), which permits the inventor to prohibit anyone else from making, using, or selling his or her invention for a specific period of time. Patents must meet certain criteria which include non-obviousness or the inventive step which requires that there be a real invention and not a simple result from some obvious extension of existing technology. Novelty, which refers to something created that is new and utility or usefulness of the invention. Patents may be related to a process, a product produced by a process.

In animal sciences, patent can be granted for a product (gene sequence, protein), process and research tool (receptors or antibodies, apparatus (PCR machine). Technologies in animal breeding that qualify for IP protection include statistical methods, DNA markers, methods to 2 53

measure traits, methods to identify animals and computer software (Ramesha et al, 2010, Matthew Rimmer, 2008).). Patents in Animal Sciences in India In India, Patents are granted for products or processes that are novel, useful, and in respect of which an inventive step can be demonstrated. Section 2 (j) of the Patents Act, 1970 defines 'invention' as any new and useful (i) Art, process, method or manner of manufacture; (ii) Machine, apparatus or other article; (iii) Substance produced by manufacture

Section 3 of the Patents Act, 1970 India, (39 of 1970, as amended by the Patents (Amendment) Act, 1999, effective from the 1st January, 1995) plays a major role in deciding the grant of patents. One has to remember here that the criteria for the grant of patent are, Novelty, Inventive Step [Non-obviousness], Industrial Applicability [Utility] and Statutory Restrictions [Sections 3 & 4]. The following are considered as non patentable inventions. 1. Plant variety 2. Animal species 3. Any part of a plant or an animal [including seeds] 4. Essential biological processes 5. Traditional knowledge component or a group of components 6. Methods of treatment of animals either to increase their value or to increase the value of the derivable products of that animal 7. Methods of treating the animals to make them disease resistant etc., 8. Bioinformatics 9. Any inventions which may be frivolous, against to the established natural laws, affecting the plant or animal life or the environment 10. Discoveries of scientific principles and thermos

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3

11. Inventions related to obscenity (any statement or act which strongly offends the prevalent morality of the time) 12. A method of agriculture and horticulture The patentable inventions 1. Genetically modified microorganisms 2. Nucleotide sequences and Amino acid sequences 3. Methods of extraction of bioactive compounds from plant or animal kingdoms. 4. Methods of making the disease resistant, drought resistant plant varieties 5. Processes of making the genetically modified plants with desirable characters. 6. Herbal extracts and methods of obtaining them 7. Cultures and methods of culturing and Nutrition mediums etc., Copyrights Copyrights are used to protect IP that includes articles, books, web pages, computer software and music. In general, copyrights involve limited formal procedures. In many cases the creative work can be copyrighted by simply adding the symbol, but registering the mark provides formal legal protection. Trademark Trademarks are protection used to identify the source of the owners' goods, products or services. As with copyright they can be informal (simply using TM) or formally registered (®). Trademarks applied to specific products that are required to be from a certain origin and have a specific level of quality (Lesser, 2002). In animal science, examples of trademarks are "Berkshire Gold™'', signifying a pork product that is Berkshire in origin and PICmarq TM signifying gene marker information used in selection of pigs sold by PIC company. Nebel et al. 2002). Trade secret A trade secret represents IP that is not revealed by the owner and therefore confers some competitive advantage. Trade secrets have no time constraints, as long as the information is kept 55

secret. Examples of trade secrets in animal breeding might include specific information to make specialized synthetic lines or specialized selection programs and indexes. More recently the definition of trade secrets has become a potential issue for the protection of seed stock. Plant Breeders’ Rights (PBR) PBR is a special purpose system only for plants. In most countries, PBR are administered by an office of the agriculture department, rather than by a separate patent office. Technically, it is a form of sui generis or special-purpose system, because of the exclusive focus on plants. PBRs are quite recent, with origins in the 1930s and 1940s. UPOV (the French acronym for the International Convention for the Protection of New Varieties of Plants), the international convention, was established only in 1961. PBRs replace utility, non-obviousness and novelty for patent systems with uniformity, stability, distinctness and novelty (FAO, 2005). CBD and TRIPS At the international level, Convention on Biological Diversity (CBD) of United Nations and Trade Related Aspects of Intellectual Property (TRIPs) Agreement of World Trade Organization are the two major agreements governing access and rights over genetic resources. As India is signatory to both CBD and TRIPs, there is a need to assess the issues at hand with a global perspective and also narrate and evaluate the options for the typical livestock production situation existing in the country (Bhat, 2009). Biological Diversity Act 2002 India is signatory to CBD and TRIPS Agreement and also approved the Global Plan of Action on Animal Genetic Resources. Therefore, it is obligatory for India to develop suitable legislation to protect and improve livestock genetic resources and associated traditional knowledge. As a member to the UN Convention on Biological Diversity, India formulated the Biological Diversity Act, 2002 which provides that the authority to determine access to genetic resources rests with the national government and is subject to national legislation. At present, information available on India's biodiversity is inadequate, including their potential use and value. Similarly, there is limited capacity and awareness regarding the implementation of the Biological Diversity Act, 2002. IPR issues related to biodiversity protection and conservation in 56

India are currently managed under the Biodiversity Act, 2002. The main objectives of the Act 2002 are conservation of biological diversity, sustainable use of its components and fair and equitable sharing of the benefits arising out of the use of biological resources and knowledge. Under TRIPS, provision is made for plant variety protection under patents or a sui generis system, or a combination of both. There is no comparable system for animals. Under TRIPS neither prior informed consent nor protection of TK is mandated. TRIPS also do not offer sufficient recourse for balancing the rights of the owners of AnGR (Ramesha, 2011).

Livestock keepers’ have the risk of losing their intellectual property rights, while biotechnology firms require a legal framework to operate. Thus, there is an urgent need for internationally recognized legally binding instrument which gives right to livestock keepers including rights over associated indigenous traditional knowledge (Damodaran, 2008). Bioprospecting and Biopiracy Bioprospecting is the exploration, extraction and screening of biodiversity and traditional knowledge for commercially valuable genetic and biochemical resources. Frequently, the term bioprospecting refers to the use of traditional knowledge in finding leads for pharmaceutical research. Biopiracy is stealing of knowledge from traditional and indigenous communities or individuals. An example is a controversy over biopiracy of the unique dwarf Indian cattle breed 'Vechur'. The unique genetic quality of this breed is the high milk fat content ranging from 6.02 to 7.86 per cent. Roslin Institute and the PPL Therapeutics (Scotland) Ltd have 14 patent applications at the European Patent Office and one of these (EP 0765390) for the gene construct of bovine alpha-lactalbumin is based on studies in Bos indicus (of which vechur is a breed). Biodiversity rich countries like India should promote bioprospecting to upgrade local capabilities and to prevent biopiracy (Ramesha et al. 2008). Need for a sui generis system for animal genetic resources In spite of significant contribution of animals to the food security and nutrition, laws and policies including intellectual property protection on genetic resources at the national as well as international level so far has focused only at plant genetic resources. There is a need for legally binding internationally accepted system for access and sharing of benefits from the use of AnGR 57

and associated traditional knowledge. There is a need for recognition of community rights over knowledge and biodiversity. Considering the special characteristics of AnGR it is essential to develop a suitable sui generi system for AnGR for protection of intellectual property rights (Ramesha et al., 2010). In case of animal breeding most of the cases superior individual animal or its germplasm is more important than providing protection for a breed/ variety. The major problem lies in identifying characteristics that could be used as a standard description of the subject matter. Breed associations and herd book registration will help in breed conservation and intellectual property protection (Verrier, 2005). A sui generis system linked to special geographical related characteristics of the animals or their products aimed at preventing misappropriation of AnGR and associated knowledge held by livestock keepers needs to be developed. Livestock keepers’ rights could include production and grazing rights as well as protection of traditional knowledge. Need for an International Treaty on Animal Genetic Resources In the modern biotechnology era, there is danger of animal genetic material from biodiversity rich countries being exploited by others, including large international companies. The development of a legal binding 'International Framework on Animal Genetic Resources' is therefore, a matter of urgency (http://dad.fao.org) on Animal Genetic Resources held at Switzerland delegates from 109 countries including India adopted a Global Plan of Action for Animal Genetic Resources, the first internationally agreed framework to halt the erosion of livestock diversity and support the sustainable use, development and conservation of animal genetic resources." In future, IP protection is likely to influence pricing and international trade of livestock products in a significant way. Livestock keepers’ have the risk of losing their intellectual property rights, while biotechnology firms require a legal framework to operate. Thus, there is an urgent need for internationally recognized legally binding instrument which gives right to livestock keepers including rights over associated indigenous traditional knowledge (FAO, 2004 and Rothschild, 2002). Conclusion To improve the productivity of livestock in India, it is absolutely necessary to increase technological interventions. Development and diffusion of knowledge intensive technologies in 58

all livestock sector are greatly influenced by IPR system. Among various types of IPR, patents, trademarks, trade secrets and geographical indications are relevant to the livestock sector. Each of these forms of IPR has different requirements and grants different legal status.

The full scope of patent protection in the field of animal breeding has not yet been determined. The application of patents to this field of innovation is fairly recent, and there is a need for more in-depth analysis of the probable consequences of introducing patents to animal breeding. Several observations are need to be made on how the special field of animal breeding calls for adaptation of the general patent laws to this particular field of technology.

Patent has been identified as a major tradable asset of enormous importance in the modern knowledge regime. Patent law has potential to alter the existing legal conditions for competition and investments in the field of animal breeding, and needs therefore a higher level of awareness among policy-makers, animal breeders and farmers.

The effects of IPRs on animal genetic resources, genetic diversity, genetic improvement and the livelihoods of livestock keepers have scarcely been analysed, but are in urgent need of exploration. Protection of native livestock breeds and associated traditional knowledge through appropriate sui generis system is essential for conservation and their sustainable use in the modern IPR system. The customary right of livestock keepers to breed their animals, which has been the driver of livestock biodiversity may be under threat unless specific national and international legal frameworks are developed. The internationally accepted legal system is need to be developed with respect to use of livestock biodiversity, then the IPR in animal sciences will bring economic benefit to the livestock keepers. Potential Patentable Areas in Animal Sciences There are unique traits in the native farm livestock species of Indian subcontinent, which can be harnessed or exploited through biotechnologies. The distinctive attributes of breeds can be exploited for profits by getting some form of Intellectual Property Rights or protection. The notable areas for IPRs could be the following in Indian context. 59

        

 Superior disease resistance.   Adaptation to withstand tropical heat and far greater climatic fluctuations.   Better potentials to utilize coarse quality feed and fodder resources.   Better survivability under more recurrent drought conditions.   Superior growth and higher fecundity in some species / breeds.   Potential to yield of certain unique products, which have economic potential after value addition.   Therapeutic and nutritional value of certain livestock products.   Geographical application of breeds associated with traits or products.   Indigenous technical knowledge on animal husbandry.   Utilization of unique genes through biotechnologies for novel use. 

With further advances in biotechnologies and processing techniques, many more IPR relevant areas would emerge. REFERENCES

Bhat Ishwara, P., (2009). The Role of Collective bodies in protection of intellectual property rights in India. Journal of Intellectual Property Rights, 14: 214-225.

Damodaran A., (2008). Traditional Knowledge, Intellectual Property Rights and Biodiversity conservation; Critical issues and Key Challenes. Journal of Intellectual Property Rights, 13: 509513.

FAO., 2004. Report by the Commission on Genetic Resources for Food and Agriculture on the Intergovernmental Technical Working Group on Animal Genetic Resources for Food and Agriculture, 2004. Food and Agriculture Organization of the United Nations, Rome.

FAO, 2005. The legal framework for the management of animal genetic resources. FAO Legislative Study. ISBN 978-82-7613-504-6, 1-33.

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Lesser W. (2002) In "Intellectual Property rights and Patenting in Animal Breeding and Genetics" Editors M.F. Rothschild and S. Newman, CAB International, axon UK, 1-16.

Matthew Rimmer. (2008). Intellectual Property and Biotechnology: Biological Inventions. Edward Elgar Publishing.

Nebel H., Akrad L., and Brady T., (2002). "Intellectual Property Rights in Animal Breeding and Genetics" Editors M.F. Rothschild and S. Newman, CABI, axon, UK pg 119-146.

Ramesha K.P., Pourouchottamane R., Kataktalware M.A. and Sarkar M., (2008). Intellectual Property Rights (IPR) issues in livestock biodiversity-Indian Perspective. Journal of Livestock Biodiversity. 1: 8-12.

Ramesha K.P., Pourouchottamane R. and Bhattacharya M., (2010). Intellectual property rightsAnimal genetic resources-Options for India. Indian Dairyman, 62: 50-63.

Ramesha K P., (2011). Intellectual Property Rights Regime for Livestock Agriculture in IndiaPresent Status and Future Prospects. Journal of Intellectual Property Rights, 16: 154-162.

Rothschild Max and Scott Newman, Intellectual Property Rights in Animal Breeding and Genetics. CABI publishing 2002.

Verrier E., Tixier-Boichard M., Bernigaud R. and Naves, M., (2005). Conservation and value of local livestock breeds: usefulness of niche products and/or adaptation to specific environments. Animal Genetic Resources Information. 36. *****************************************

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TOPIC: MOLECULAR TYPING AND CHANGING EPIDEMIOLOGY OF DENGUE IN TAMILNADU Authors: Ms. P. Padmapriya, Mr. V. Senthil kumar, Mr. R. Senthil Raja, Dr. A.K. Sheriff, Dr.T.S. Uma, Ms.C.P. Anupama, Dr. K. Kaveri*, Dr. P. Gunasekaran Name of the Institution – King Institute of Preventive Medicine & Research, Guindy, Chennai

Background: Dengue virus infection is now recognized as one of the most important vector borne human infections. The global incidence of the dengue infection has now increased enormously and an estimated 50–100 million cases of dengue infections are now reported annually from more than 100 tropical and sub tropical countries of the world (1). Dengue viruses (genus Flavivirus, family Flaviviridae) are transmitted by mosquitoes and the principal vector (Aedes aegypti) is a day-biting domestic mosquito of public importance that breeds in natural or artificial water. Dengue illnesses are caused by any of the four serologically related viruses designated as DENV-1, DENV-2, DENV-3 and DENV-4 (2). Infection with any one of these serotypes mostly causes a mild, self limiting febrile illness (classical dengue fever (DF).However, a few cases develop severe life threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Several hypotheses, like antibody dependent enhancement (ADE) in heterotypic secondary dengue infections and host factors have been suggested to explain the mechanism of pathogenesis of DHF and DSS. The number of DHF and DSS cases has been increased in India. Occurrence of concurrent infection has been implicated as the causative agent in most of these outbreaks (3).

Laboratory diagnosis of dengue infection is primarily achieved through virus isolation from acute phase serum, serodiagnosis and molecular assays for detection of viral RNA. Virus isolation though considered 'gold standard' is technically demanding and time consuming. The most rapid serological technique such as IgM ELISA with a single serum sample, does not furnish information about the serotype of the virus. The plaque reduction neutralization technique (PRNT) allows typing but is costly, slow and difficult to perform (4). The molecular methods based on PCR technique offer a suitable alternative to conventional isolation technique. The RT-PCR targeting the conserved regions of C-prM gene junction is widely employed for 62

precise confirmation of dengue infection. In the present study, we report the serological and molecular investigations among suspected dengue cases (5). In recent years co-circulation of multiple dengue virus serotypes is being increasingly reported with concurrent infections. However the association of concurrent infections with severe forms of disease (DHF/DSS) needs further studies. From India there are few studies reporting cases of dual infection (6). In this study we report the results of the detection of dengue virus RNA by RT-PCR during the year 2014. The main feature of this report is the co-circulation of all the serotypes of dengue virus and to highlight percentage occurrence of concurrent infections probably leading to DHF/DSS.

Materials & Methods: The study was performed at the King Institute of Preventive Medicine, Department of Virology, Guindy. Samples were received from Government General Hospital, Institute of Child Health, other Government hospitals and private institutions all over Chennai. Since these are tertiary care centres, the samples received were inclusive of referred patients from the various districts in Tamil Nadu. Both adult and paediatric cases were included in this study. The patients were referred to as suspected DF cases, based on standard diagnostic criteria (7). Most of the samples were collected during 5 to 10 days of illness. Approximately 2-5 ml of blood was received, serum separated and subjected to ELISA. MAC ELISA was performed using kit from NIV, IgM Capture ELISA and IgG ELISA was also performed using kits from Panbio (Australia). Few samples were tested for the presence of NS1 Antigen. Positive samples were further subjected to RT-PCR. Viral RNA extraction was performed using Qiagen Viral RNA kit. Published primers by Lanciotti et al were used in this study for PCR (8). Controls were put up where ever necessary.

Statistical analysis: The data presented were analyzed using Chi-square test for proportion and the Chi-square test for linear trend using the Graphpad prism 6.01 programme.

63

RESULTS: During the study period, the total number of samples screened was 1249, of which 180 (14.4%) were positive for IgM antibodies. Of these 180 samples 49 (27.2%) were positive for the presence of NS1 Antigen. 70 (38.8%) samples were IgG positive. For the present study the NS1 positive samples were further analysed. An age wise distribution was performed. RT PCR positive were observed in both pediatric and adult cases. Few samples were both NS1 and IgG positive leading to probable secondary infection. Table 1 depicts the distribution of RT PCR positive samples in the pediatric age group. It was observed that 2 samples in the 0-5 age group and 6-12 was found to be both RT-PCR positive and IgG positive confirms the presence of secondary serotype.

Table 1: Pediatric Age Group NS1 TOTAL PEDIATRIC POSITIVE

19

0-5 IgM

NS1 9

6-12 IgM

IGG 1

RT 2*

NS1 10

IGG 1

RT 3**

*1 sample was positive for both dengue 1&4 **2 sample was positive for both dengue 1&3

Table 2: Adult Age Group 18-Dec TOTAL ADULT POSITI VE

30

19-30

31-45

31-45

46-60

60>

N IG S IG NS IG R NS IG R NS IG R NS IG R NS1 G RT 1 G RT 1 G T 1 G T 1 G T 1 G T 6 1 2* 7 1 3** 5 0 2 4 1 1 4 0 1 4 0

*1 sample was positive for both dengue 1&4 **1 sample was positive for both dengue 1&3 Table 2 depicts the distribution of RT PCR positives among the Adult population. It was found that adult in population there were presence of secondary serotype.

64

Figure 1: Depicts the gel picture of positive samples showing the presence of dengue 1&3 in one sample, dengue 2 in one sample and dengue 1 &4 in one sample. For each sample 4 wells were taken up.

DISCUSSION: The changes in the global epidemiology of dengue during the last 50 years lead to the increase in the number of countries reporting dengue. The first epidemic of dengue in India occurred in Kolkata in 1963–64 and ever since the epidemiology of dengue virus has been changing (9). As reported by Preeti Bharaj et al., dengue-1 and dengue-3 were identified to be the most frequent dengue virus serotype combination occurring during this outbreak. This was followed by the dengue-1 and dengue-4 serotype combination (10). Our study implicated that dengue 1&3 combination followed by dengue 1&4 was commonly observed in Tamilnadu. It has been postulated that concurrent infections by multiple dengue virus serotypes may influence the clinical expression of the disease (11). This is considered as a single major factor for the emergence of DHF. Presence of combination of serotypes in the Pediatric age group confirms us the presence of secondary infection. Dengue 2 as a single serotype is known to be the cause of complications leading to DHF and DSS (12). The transmission of dengue increases in monsoon, as was also observed in our study. This shows that the presence of stagnating water after rainfall favour breeding of mosquito vector resulting in an increased incidence of dengue. These findings also indicate that preventive measures against dengue infection should probably come into full swing during the monsoon and post monsoon months (13). The present study reports the emergence of multiple serotypes circulating in Tamilnadu. Further studies regarding the molecular characterization of these dengue viruses are underway. Epidemiology of dengue infection in Tamilnadu is rapidly changing face, with frequency of outbreaks increasing, even as dengue establishes itself as endemic disease(14). The need of the hour is to characterize the circulating serotypes of dengue virus in our community and understand the evolutionary processes influencing the dengue virus, as this is expected to impact on vaccination strategies for future (15).

65

CONCLUSION: The increasing trend of co-circulation of dengue virus serotypes suggests that Tamilnadu is becoming a hyperendemic state from an endemic one. Although the sample size in this study was small, the study highlights a high percentage of concurrent infections by different dengue virus serotypes.

REFERENCES 1. World Health Organization: Dengue and dengue haemorrhagic fever. Fact sheet 2002, 117 2. Henchal EA, Putnak JR: The dengue viruses. Clin Microbiol Rev1990, 3:376-96. 3. Mc Bride WJH, Bielefeldt-Ohmann H: Dengue viral infections;pathogenesis and epidemiology. Microbes and Infect 2000,2:1041-50. 4. Gubler DJ: Epidemic dengue/dengue haemorrhagic fever: a global public health problem in the 21st century. Dengue Bulletin 1997, 21:1-15. 5. World Health Organization: Dengue and dengue hemorrhagic fever. . Accessed 1 July 2007. 6. Gubler DJ: Dengue and dengue hemorrhagic fever: its history and resurgence as a global public health problem. In Dengue and dengue hemorrhagic fever Edited by: Gubler DJ, Kuno G. London: CAB International; 1997:1-22. 6. Gubler DJ: Epidemic dengue/dengue haemorrhagic fever as a public health, social and economic problem in the 21st century. Trends in Microbiology 2002, 10:100-103. 7. WHO: Dengue: Guidelines for diagnosis, treatment, prevention and control. New edition. Geneva: WHO; 2009. 8. Gubler DJ: The arboviruses: epidemiology and ecology. In Dengue Volume II. Edited by: Monath TP. CRC Press, Boca Raton, FL;1988:223-260. 9. Gubler DJ: Dengue and Dengue hemorrhagic fever. Clin Microbiol Rev 1998, 11:480496 10. Concurrent infections by all four dengue virus serotypes during an outbreak of dengue in 2006 in Delhi, India. Preeti Bharaj1, Harendra S Chahar1, Anubhav Pandey1, Kavita Diddi1, Lalit Dar1, Randeep Guleria2, Sushil K Kabra3 and Shobha Broor. Virology Journal 2008, 5:1 11. Guzman MG, Kouri G: Advances in dengue diagnosis. Clin Diagn Lab Immunol 1996, 3:621-7. 66

12. Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, Vorndam AV: Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptasepolymerase chain reaction. J Clin Microbiol 1992, 30:545-51. 13. Laille M, Deubel V, Sinte-Marie FF: Demonstration of concurrent dengue 1 and dengue 3 infection in six patients by the polymerase chain reaction. J Med Virol 1991, 34:51-54. 14. Maneekarn N, Morita K, Tanaka M, Igarashi A, Usawattanakul W, Sirisanthana V, Innis BL, Sittisombut N, Nisalak A, Nimmanitya S: Applications of polymerase chain reaction for identification of dengue viruses isolated from patient sera. Microbiol Immunol 1993, 37:41-47. 15. Kanesa-Thasan N, Chang GJ, Smoak BL, Magill A, Burrous MJ, Hoke CH Jr: Molecular and epidemiologic analysis of dengue virus isolates from Somalia. Emerg Infect Dis 1998, 4(2):299-303.

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Evaluation of anticancer activity of green tea extracts in Human epithelioma of larynx (Hep-2) cell line. N.Saran1, G.Bupesh1, S.Magesh1, B. Anandharaj2, P.Gunasekaran1 C.P.Anupama1, A.K.Sheriff1 and K.Kaveri1 1

Department of Virology, King Institute of Preventive Medicine and Research, Guindy, Chennai, India. 2

Department of Microbiology, M.R. Govt. College, Mannargudi, India.

Abstract Green tea is rich in all the antioxidants. It has rejuvenating activity in our day to day life. The daily intake of green tea consumption per person was assessed as 2gm/day approximately. In this lieu, the present study was inducted to evaluate the anticancer efficacy of green tea against Human epithelioma of larynx cancer cell line. Ethanolic, chloroform and aqueous extracts of green tea were evaluated for the anticancer activity at various concentrations from 10µg to 2mg. The progressions of Hep-2 cells were periodically evaluated for 24, 48 and 72 hours. It was observed from the study that the aqueous extracts showed potent anticancer activity at minimum concentration of 50 µg than the other two extracts whereas the ethanolic and chloroform extracts showed anticancer activity at 2mg concentration. The phytochemistry profiles of green tea revealed the presence of polyphenols and flavonoids. The anticancer activity of green tea is due to the presence of bioactive secondary metabolites (polyphenols and flavonoids). Keywords: Green tea, Hep-2 cells, Anticancer activity and Bioactive compounds. Corresponding Author; Email; [email protected],[email protected] Introduction: Plants such as aromatic plants, herbs and spices are known natural resources that have been employed to produce highly nutritious and valued products. The products such as food, cosmetics, nutritional and pharmaceutical industries. Though their healing properties and health68

promoting effects have not yet been fully elucidate, several traditional herbal medicinal products have long been used to heal and cure diseases and to improve health. There are antimicrobial drugs for the treatment of inflammatory and infectious diseases were now a days not effective for therapeutics. In this lieu the available drugs have recently revived and the search for natural substances and compounds with antimicrobial properties (Jain et al., 2010), including medicinal plants (Bauer et al., 1966) were lead to clinical research. Tea is the one of the second most commonly consumed liquid after water and the most common beverage drunk. Tea, the drink, is an infusion of variously processed leaves and flowers of one of the varieties of an evergreen shrub botanically called Camellia sinensis. Green tea (Camillia sinensis L.) is prepared from the steamed and dried leaves of an evergreen shrub native to eastern Asia. It is consumed mainly in Japan, China and other Asian Countries including India(Kuroda Y & Hara Y (1999). Green tea, the minimally fermented ( oxidized) product of the tea leaf, may show certain health benefits. In addition to their traditional use for making tea, the leaves of Camellia sinensis L. are also industrially processed. Many natural substances have been identified in green tea; green tea components theanine and catechins have neuroprotective effects( Yokogoshi H & Kobayashi M(1998),Kakuda T(2007). It has significant role in cancer prevention. Green tea catechins have been shown to inhibit tumor cell proliferation and promote destruction of leukemia cells( Smith D M & Dou Q P(2001) and breast cancer cells(Vergote D et al(2002) , Masuda M et al(2002). In the present study, the phytochemistry and anti cancer efficacy of green tea extracts were investigated in Hep-2 cells (Human epithelioma of larynx). Materials and Methods: Hep-2 cells (Human epithelioma of larynx) Cell lines and culture condition Hep-2 cells (Human epithelioma of larynx) were used to investigate the cytotoxicity effect of essential oil. This cell lines were grown in RPMI 1640 media (Gibco) supplemented with 10% (v/v) foetal calf serum (FCS) and 2 mM L-glutamin in tissue culture flasks (Nunc). The media were changed twice a week and kept at 37°C in a humidified atmosphere of 95% air and 5% CO2. Chemicals and consumables

69

MEM culture medium, antibiotics – antimycotic solution, fetal bovine serum (FBS) and trypsin were purchased from Invitogen, Life Sciences, USA. Consumables and culture wares used in the study were procured from Nunc, Denmark. Ethanol and all other specified reagents and solvents were purchased from Sigma Chemical Company Pvt. Ltd. St. Louis, MO, USA. Collection of plant material The fresh leaves of Camellia sinensis were collected by following (Tariq and Reyaz, 2012) from the dense tea state garden at Ooty, Coimbatore district, Tamil Nadu South India. The leaves were shade dried and then dried at 70oC in hot air oven. The dried leaves were then ground to powder using mortar and pestle. The powder is stored in airtight closed bottles until use. Aqueous extraction 10g of each of the ground leaves were extracted by soaking for 2 days using 100ml of distilled water in a 250ml sterile conical flask. The extracts were filtered using Whatman filter paper No 1. The filtrates were then concentrated by using rotavapour and stored in universal bottles and refrigerated at 4°C prior to use. Aqueous extraction was carried on as preliminary study. Ethanolic extraction 10g powdered samples of each were extracted with 100 ml of Ethanol in a successive manner to produce crude extracts containing wide range of active compounds. The extracts were prepared by maceration of the plant material with the solvents in a shaker for 2 d. The respective extracts were filtered using Whatman No.1 filter paper and dried under reduced pressure at a temperature below 45°C in rotavapour to yield a dense residue. Chloroform Extraction 10g powdered samples of each were extracted with 100 ml of Chloroform solvent in a successive manner to produce crude extracts containing wide range of active compounds. The extracts were prepared by maceration of the plant material with the solvents in a shaker for 2 d. The respective extracts were filtered using Whatman No.1 filter paper and dried under reduced pressure at a temperature below 45°C in rotavapour to yield a dense residue. 70

MTT test The proliferation rates of Hep-2 cells after treatment with essential oil were determined using the colorimetric 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The yellow compound MTT was reduced by mitochondrial dehydrogenases to the waterinsoluble blue compound formazan, depending on the viability of cells. Phytochemical screening Preliminary phytochemical screening of the extracts about 2g of the dried extract was weighed and dissolved in distilled water. The solution obtained was then subjected to various phytochemical screening to identify the chemical components present in the extract using qualitative chemical tests with the use of chemical reagents. Standard methods were employed for testing carbohydrates, tannins, saponins, flavonoids, alkaloids, steroids and terpenes.(Trease and Evans 2002) and (Sofowora 1982). Results and Discussion Green tea (Camellia sinensis) extracts of ethanolic, chloroform and aqueous extracts were screened for the phytochemical profiles for the bioactive secondary metabolites. The qualitative analysis of phytochemicals on the different extract showed the presence of active metabolites such as, flavonoids, steroids, gallic tannins, terpenoid, saponins, and glycosides. The table .1 shows the presence of the secondary metabolites in the different extracts. Then the anticancer efficacy of Camellia sinensis were evaluated in the Hep-2 cells (Human epithelioma of larynx). Concentration of Camellia sinensis 10µg to 2mg at different time intervals such as 24, 48 and 72 hours were periodically evaluated. The anticancer activity of Camellia sinensis were presented in the figure-1. The anticancer activity of C.sinensis aqueous extracts showed potent activity at 50 µg concentration where as the ethanolic and chloroform had anticancer activity at 2mg concentration. The aqueous extract showed toxicity at 72 hours duration in the figure-1. But the rest of the two extracts displayed activity at 24 hours intervals. The higher toxicity with respect to time intervals was significantly observed in the Hep-2 cells. The figure-2 showed antiproliferative activity by MTT cytotoxicity assay. Aqueous extract at Concentration of 50 µg indicate the 50% inhibition than the rest of the concentrations. The ethanolic and

71

chloroform extracts at Concentration of 2mg reveals the 50% inhibition of proliferation on Hep-2 cells. Table.1 Phytochemical Profiles of Green tea Camellia sinensis extracts Secondary Metabolites

Aqueous Ethanol

Chloroform

Tannins

+

-

+

Saponins

+

-

+

Cardiac Glycosides

-

-

+

Terpenoids

-

+

-

Flavonoids

+

+

-

Polyphenols

+

-

-

Figure-1 Anticancer activity of Camellia sinensis extracts on Hep-2 cells (Human epithelioma of larynx)

( A. Control Hep-2 cells (Human epithelioma of larynx) cells, B. Aqueous extract of C. sinensis 750 µg at 72hrs, C. Aqueous extract of C. sinensis 2 mg at 72hrs D. Chloroform extract of sinensis 2mg at 72hrs, E. Ethanolic extracts of C. sinensis 2 mg at 48 hrs, F. Ethanolic extracts of C. sinensis 2 mg at 72hrs ) 72

Antiproliferative activity

Figure-2 Antiproliferative activity of Green tea extracts in C. sinensis 500.00 450.00 400.00 350.00 300.00 250.00 200.00 150.00 100.00 50.00 0.00

Aqeous Ethanolic Chloroform

10

25

50

100

250

500

750

1000

2000

Green tea Concentration in µg The aqueous extracts of

R. officinalis has also been demonstrated the cytotoxicity

activity in hela cells (Khafagi et al., 2000; El-Meleigy et al., 2010). Similarly the aqueous extract of green tea showed higher cytotoxicity in the Hep-2 cells than the ethanolic and chloroform extracts. The cytotoxicity of green tea was presented in the figure-2. Conclusion The phytochemistry analysis of C. sinensis revealed the presence of Tannins,Saponins, Terpenoids, Flavonoids and polyphenols. The green tea possesses potent anticancer properties against (Hep-2) Human epithelioma of larynx cells. The screening study revealed that the three solvent extracts were found to be a inhibitory to cell proliferation against Hep-2 cancer cell line. Further experimental analysis on these plants would definitely reveal the above bioactive secondary metabolites were highly responsible for cancer cell death due to the inhibitions of active principles or chemical constituents. REFERENCES 1. Kuroda Y & Hara Y, Antimutagenic and anticarcinogenic activity of tea polphenols, Mutat Res, 436 (1999)69. 2. Yokogoshi H & Kobayashi M,Hypertensive effect of gammaglutamylmethylamide in spontaneously hypertensive rats,Life Sci,62 (1998) 1065.

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3. Kakuda T,Neuroprotective effectsof the green tea components theanine and catechins, Biol Pharm Bull, 25 (2007) 1513. 4. Smith D M & Dou Q P, Green tea polyphenols epigallocatechin inhibits DNA replication and consequently induces leukemia cell apoptosis, Int J Mol Med, 7(2001) 645. 5. Vergote D,Cren-Olive C,Chopin V, Toillon R A, Rolando C,Hondermarck H & Le Bourhis X, (-)-Epigallocatechin (ECG) of green tea induces apoptosis of human breast cancer cells but not of their normal counterparts, Breast Cancer Res, 76 (2002) 195. 6. Masuda M,Suzui M,L IM J T,Deguchi A,Soh J W & Weistein I B,Epigallocatechin-3gallete decreases VEGF production in head and neck and breast carcinoma cells by inhibiting EGFR-related pathways of signal transduction, J Exp Ther Oncol, 2 (2002) 350. 7. Trease GE, Evans WC. Introduction and General Methods in Pharmacognosy. 14th ed. London: Alden press; 2002. 8. Sofowora EA. Traditional medicine and medicinal plant of Africa. Nigeria: Spectrum books publishers; 1982. 9.

Okubo S, Ikigai H, Toda M, Shimamura T. The anti-haemolysis activity of tea and coffee. Lett Appl Microbiol 1989; 9(2):65-6. *******************

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