Vidya (2019) Vol : 14(2)

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From the Editorial Desk

Carried out with the help of GSLV Mk-III launch vehicle, Chandrayaan 2 was launched from the Satish Dhawan Space Centre in Sriharikota, Andhra Pradesh.

India will become the fourth country to land a spacecraft on the Moon. So far, all the landings have been in the areas close to the Moon’s equator. Chandrayaan-2 will make a landing at a site where no earlier mission has gone, i.e., near the South Pole of the Moon. It can contain clues to the fossil records of early Solar System. The primary objective of Chandrayaan-2 is to demonstrate the ability to soft-land on the lunar surface and operate a robotic rover on the surface. Scientific goals include studies of lunar topography, mineralogy, elemental abundance, the lunar exosphere, and signatures of hydroxyl and water ice.

The unexplored territory gives an opportunity for the Mission to discover something new. The South Pole of the Moon holds possibility of presence of water. In addition, this area is also supposed to have ancient rocks and craters that can offer indications of history of the Moon. The launch vehicle will be carrying an orbiter, a rover named Pragyan and a lander named Vikram.

And what makes this mission even more special is that the Project Director and Mission Director of the Rs 978 crore Chandrayaan-2 are both women scientists from the Indian Space Research Organisation (ISRO). Also, it is noteworthy that 30 percent of the team leading Chandrayaan-2 are all women. Who can forget the team of women scientists rejoicing at the success of the Mars Orbiter Mission (MOM) in 2014? Ritu Karidhal is the Mission Director of Chandrayaan-2. M Vanitha, the Project Director of Chandrayaan-2, have been working with ISRO for a long time.

India creates history with the launch of Chandrayan 2.

Prof. (Dr.) Meenu Saraf

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Original Paper ISSN: 2321-1520

Solid Waste Management Survey in Kankaria Lake, Ahmedabad

Munema Suratwala1, Jaimini Kumarkhaniya1, Honey Panchal1, Vinit Damor1 and Hitesh Solanki2*

1. Student, Department of Environmental Science, USSC, Gujarat University, Ahmedabad, Gujarat 2. Professor and Head, Department of Environmental Science, USSC, Gujarat University, Ahmedabad, Gujarat

E-mail: [email protected]

Received Date: 2-7-2019 Published Date: 15-9-2019

Abstract

Municipal solid waste management is one of the major environmental problems of Indian cities. Improper management of municipal solid waste (MSW) causes hazards to the inhabitants. Various studies reveal that 90% of MSW is disposed of unscientifically in open dumps and landfills, creating problems in public health and the environment. The Kankaria Lake is a well-known place in Ahmedabad. This paper presents an assessment of the existing situation of municipal solid waste management in Kankaria. Zoo, Butterfly-garden, Kids park and Nagina Vadi are comprised in this research. Every aspect of solid waste management related to the Kankaria Lake is covered in this paper.

Keywords: Solid waste management, Kankaria-lake, Zoo.

Introduction

Rapid population growth, urbanization and industrial growth have led to severe waste management problems in the cities of developing countries like India. India is an agriculturally based country 3 Vidya (2019) Vol : 14(2) Solanki et al

with a present population of approximately 1.2 billion (Union Health Ministry, 2011). According to the provisional figures of Census of India 2011, 377 million people live in the urban areas of the country. This is 31.16 % of the Country’s total population. There are 29 states and seven union territories in the country. India has led to the migration of people from villages to cities, which generate thousands of tons of MSW daily. The Municipal solid waste (MSW) amount is expected to increase significantly in the near future as the country strives to attain an industrialized nation status by the year 2020 (Sharma and Shah, 2005; CPCB, 2004; Shekdaret al., 1992). Poor collection and inadequate transportation are responsible for the accumulation of MSW at every nook and corner. The management of MSW is going through a critical phase, due to the unavailability of suitable facilities to treat and dispose of the larger amount of MSW generated in metropolitan cities. Unscientific disposal causes an adverse impact on all components of the environment and human health (Rathi, 2006; Sharholyet al., 2005; Ray et al., 2005; Jhaet al., 2003; Kansal, 2002; Kansalet al., 1998; Singh and Singh, 1998; Guptaet al., 1998).

Generally, MSW is disposed in low-lying areas without taking any precautions or operational controls. Therefore, MSWM is one of the major environmental problems of Indian Megacities. It involves activities associated with generation, storage, collection, transfer and transport, processing and disposal of solid wastes. But in most cities, the MSWM system comprises of only four activities, i.e., waste generation, collection, transportation, and disposal. The management of MSW requires proper infrastructure, maintenance and upgradation of all activities. This becomes increasingly expensive and complex due to the continuous and unplanned growth of urban centers. The difficulties in providing the desired level of public service in the urban centers are often attributed to the poor financial status of the managing municipal corporations (Moret al., 2006; Siddiquiet al., 2006; Rajeet al., 2001; MoEF, 2000; Ahsan, 1999). During the last three decades, the National Environmental Engineering Research Institute (NEERI) has carried out studies in more than 50 cities and towns in India. Characterization of MSW indicated that the waste consists of 30-45% organic matter, 6-10% recyclable and the rest as inert matter. The organic matter in solid waste in developing countries is much higher than that in the developed countries (Bhide and Sundersan, 1983), and organic matter can be converted into useful products to reduce the burden on existing landfills (Richard, 1992). The MSW (Management and Handling) Rules, 2000 recommend source-

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specific waste collection and transportation in addition to appropriate processing and disposal. In this paper, it is shown that how much of solid waste is generated from the Kankaria-Lake.

Methodology

Survey based methodology is adopted for the present study.

Result and Discussion

In Kankaria most of the waste is organic matter as it is surrounded by so many trees. The paper and plastic waste is in lower amounts than leaves and sticks. The cleaning of Kankaria starts at night around 8.00 pm. everyday. There are total 64 to 68 sweepers are working under the Ahmedabad Municipal Corporation. All of them are hired by a private company to take care of the Kankaria-lake. This cleaning ends at 5.00 am. in the next day morning. There are three shifts, morning, evening and night for cleaning and disposing the solid waste from whole place. It consists the waste from the Zoo, Food-Stalls, Kids-Park, Periphery, Butterfly Park and surrounding of the walls of Kankaria.

In the Kamla Nehru Zoological Garden, the waste is utilized in making manure for plants. Zoo generates a very less amount of paper and plastic waste. In manure plant 600 ml of microbial liquid and 200 liter of water is used for making the manure. All the waste is mixed with microbial liquid and water very well in the rotator. After rotating this mixture, it is placed for aerobic process near by area. After 15 days this mixture is screened on the vertical screening machine. After the screening the larger parts are used again for aerobic process and the screened small particles are collected in the big bag. This manure production plant used 100 m2 of land for manure production. This process require75 kWh of electricity. The manure plant is near the gate of the zoo and it produces more than 1000 kg manure in 3-4 days. This manure is sent to the Ahmedabad Urban Development Authority (AUDA) gardens in the Ahmedabad city for using it as fertilizer for plants.

Plastic and paper waste are send to the industries for recycling once in a week. The workers from industries come to Kankaria to collect the plastic bottles. They segregate them and then carry these bottles to plastic recycling industry. The food waste is collected in different blue coloured bags. They are kept separate for disposal. Whatever the green waste is generated/collected from the periphery of the Kankaria-lake is sent to small part called bore well station in the Kankaria. 5 Vidya (2019) Vol : 14(2) Solanki et al

Workers use the sticks as wood-fuel for cooking and other purpose, who live there, and the rest of the plant leaves and sticks are sent to compost making industries.

The Kankaria Lake area generates 7000 kg to 8000 kg of solid waste on a daily basis. The amount increases when it is a national holiday and on weekend days. It increases up to 12000 kg to 14000 kg during holidays. All the leftover solid waste is disposed to the outside of Kankaria which is later collected by AMC trucks and transported to Pirana, the dumping site of Ahmedabad city.

Name of Holidays and numbers of Public entry at Kankaria as follows: Rakshabandhan – 38974, Indepedence Day - 53168, Janmashthami – 8093, Diwali – 54000, Hindu New year – 68000, Bhaiduj – 77000. As per the record of the Kankaria lake administration, these are the entries of people on Sundays and national holidays for the month of August to November 2017.

Figure shows percentage of waste generated during holidays.

Conclusion

Solid waste management is a big challenge in bigger cities nowadays. As the research has done in Kankaria, it really looks very difficult to manage all the prospects related to waste management. Some places lacks advanced equipment and modern ideology.

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The disturbing thing which was found in every part of research is lack of awareness in the public. We the people of Ahmedabad or as citizens of India need to be aware of this situation first. We must understand the growing problem of waste disposal. We must propagate the recyclable/reusable things in first demand. We must stop using plastic bags at a certain point. Awareness is the only way to better and sustainable future.

Acknowledgement

We would like to thank Ahmedabad Municipal Corporation and The Kankaria Zoo authorities for providing us necessary facilities and permission.

References

Gupta Shuchi, Krishna Mohan, Rajkumar Prasad, Sujata Gupta and

Arun Kansal (1998). Solid waste management in India: options andopportunities. Resources Conservation and Recycling 24(2):137

ManjuRawat, AL. Ramanathan, T. Kuriakose (2013). Characterisation of Municipal Solid Waste Compost (MSWC) from Selected Indian Cities — A Case Study for

its Sustainable Utilisation. Journal of Environmental Protection, 4(2): 163-171. doi: 10.4236/ jep.2013.42019.

Neha Gupta, Krishna Kumar Yadav, Vinit Kumar (2015). A review on current status of municipal solid wastemanagement in India.Journal of Environmental Sciences.37:206-217 DOI: 10.1016/ j.jes.2015.01.034.

Rajendra Kumar Kaushal, George K. Varghese , Mayuri Chabukdhara An insightArticle 2012: Municipal Solid Waste Management in India Current State and Future Challenges. International Journal of Engineering Science and Technology. 4(4): 1473-1489.

SharholyMufeed, KafeelAhmad , Gauhar Mahmood and R.C. Trivedi (2008). Municipal solid waste management in Indian cities – A review.Waste Management 28: 459–467.

Sudhira, V., G. Srinivasana and V. R. Muraleedharana (1997).Planning for sustainable solid waste management in urban India.System Dynamics Review 13 (3): 223–246 John Wiley & Sons, Ltd.

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CCC 0883-7066/97/030223–24 $17.50

Sunil Kumar, J.K. Bhattacharyya, A.N. Vaidya, Tapan Chakrabarti, Sukumar Devotta, A.B. Akolkar (2009). Assessment of the status of municipal solid waste management in metro cities, state capitals, class I cities, and class II towns in India. Waste Manag. 29(2):883-95. doi: 10.1016/ j.wasman.2008.04.011.

TapanNarayana,(2009). Municipal solid waste management in India: From waste disposal to recovery of resources.Waste Manag. 29(3):1163-6. doi: 10.1016/j.wasman.2008.06.038.

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Research Paper

ISSN: 2321-1520

Synthesis of Reduced Graphene Oxide Using Tea Extract and Its Application in the Removal of Methylene Blue from Effluents.

Himanshu V Madhad, Dilip V Vasava*

Chemistry Department, School of Sciences, Gujarat University, Navrangpura, Ahmedabad, Gujarat-380009, India. E-mail: [email protected]

*Corresponding Author

Received Date: 20-7-2019

Published Date: 15-9-2019

Abstract

An environment friendly, simple and cost-effective method for reduction of graphene oxide (GO)by using green tea extract method is reported. Reduced graphene oxide (RGO) was characterized by X-ray diffraction (XRD), Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and UV-visible spectroscopy analysis. RGO has shown to possess remarkable dye disposal property of Methylene blue dye. The Green tea reduced GO material offered a good potential in the removal of dye waste water.

Keywords: Dye adsorbent, Green Reduction, Graphene Oxide, Reduced Graphene Oxide.

Introduction

Graphene is a single layer of sp2 hybridized carbon atoms arranged in a honeycomb lattice, which has established significant attention in the scientific community worldwide due to its unique properties

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such as electrical, optical, mechanical, thermal, gas barrier, sensors, energy storage and biomedical etc. Recently, numerous scientific reports have described the application of graphene as a good adsorbent. Graphene and its derivatives have opened a new dimension for development of adsorbents for elimination of heavy metals and dyes from wastewater. Numbers of green methods have been reported for the reduction of GO with various green reagents such as alkaloids, amino acids, proteins, polyphenolic compounds, polysaccharides, enzymes and vitamins. While the reduction effectiveness found through green methods is typically lower than that obtained with chemical methods. However, application of such toxic chemicals and reducing agents for large scale remedial applications is possibly harmful to ecosystem and human health. Therefore, to minimize environmental effects, the production of eco-friendly reducing agents has been investigated. So, One and only alternative is the green synthesis of materials which have been broadly used in environmental remediation. In this work, we proposed environmentally friendly method for the reduction of graphene oxide to reduced graphene oxide (GO) by using Green tea extract. The GO were synthesized by chemical exfoliation technique and then reduced by green tea extract solution. The synthesized reduced graphene oxide (RGO) was studied to evaluate its ability to adsorb the methylene blue dye. Experimental Materials Graphite fine powder (150 mesh, 99.5%) and Sodium nitrate (NaNO3) were purchased from CDH. Potassium permanganate (KMnO4, 99.5%) was purchased from Merck. Sulfuric acid (H2SO4, 98%) and Hydrogen peroxide (H2O2,30%) were purchased from Finar Chemicals. Green tea powder was purchased from Apollo Pharmacy. All the chemicals were of analytical grade and used without further purification. Synthesis of Graphene Oxide Graphene oxide from graphite was prepared according to modified Hummer’s method.1 g of graphite powder and 0.5 g NaNO3 were added to 23 mL of concentrated H2SO4 in a 500 mL flask. The prepared mixture was stirred in an ice bath. Stir it till homogeneous paste is formed. 3 g KMnO4 was then added slowly (in little portions) with continues stirring. Then, the flask was

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removed from ice bath to maintain the reaction mixture at room temperature. After 2 hr,46 mL of deionized water was added drop wise in the reaction mixture and temperature was kept to 98 0C using an oil bath for 15 min. Then add 100ml of deionized water and 100 mL of 30% H2O2 solution into the reaction mixture. The reaction mixture turned bright yellow. Allow it to stand and decant the supernatant. The supernatant was washed with deionized water and HCl for several times. The residue was centrifuged at 8000 rpm till the pH becomes nearly 5 to 6. The residue was dried in an oven at 50 0C for 72 h. The brown coloured material was obtained. Synthesis of Green Tea Reduced Graphene Oxide.

Reduced graphene Oxide was prepared by reduction using Green tea extract. Green tea extract was used as a reducing agent because it’s an excellent source of polyphenolic compounds. 1 g of tea powder was added to 100 mL of deionized water and boiled at 100 0C for 1 h. After 1 h allow the reaction mixture to attain the room temperature. Then filter the solution. 50 mg GO was added in the solution of green tea extract and heated in an oil bath at 85 °C in a nitrogen atmosphere for 4 h. The obtained solution was filtered and washed several times with ethanol and deionized water to remove excess of polyphenols. The residue was dried in an oven at 50 0C for 24 h. The black coloured product was obtained.

Removal of Methylene blue by RGO.

Exactly 20 mg of reduced graphene oxide added in 100 ml of the methylene blue dye solutions with constant stirring. During the experiment samples were collected at specific time intervals (10, 20, 30, 40, 50 min) for the UV- vis analysis. The absorbance was recorded for methylene blue at 664 nm. The percentage removal of the dye was determined by using the equation given below.

% of removal = (Ci × Cf) ×100 Ci

Where, Ci and Cf are initial and final dye concentrations respectively.

Characterization

GO and RGO were characterized using X-ray diffraction (XPERT-PRO diffractometer) equipped with goniometer PW 3050/60 working with Cu(Ka) radiation of wavelength 1.5406 A° , Scanning

11 Vidya (2019) Vol : 14(2) Vasava et al electron microscopy (SEM, Hitachi S-3000N), Fourier transform infrared spectra frequency range of 600 to 4000 cm-1 by Bruker FT-IR spectrometer and UV- vis absorption spectra a wavelength range of 200–800 nm by Jasco model-v630 spectrometer.

Result and Discussion

X-ray diffraction pattern (Fig. 1), the appearance of 2? broad peak at 26.50 2 theta is due to the formation of few layers of RGO sheets. The reflection peak appeared at 100 2 theta value was attributed to GO. The peak of GO which was absent in the diffraction pattern of RGO suggesting the absolute exfoliation of GO. Another peak at 430 2? coincide to the turbostratic band of disordered RGO.

Fig. 1. XRD pattern of RGO

SEM studies (Fig. 2) reveled that the formation of transparent and exfoliated RGO due to the procedure of reduction with green tea extract.

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Fig. 2. SEM image of RGO

The peak in UV-visible spectrum at 234 nm is accredited to the p–p* transition of C=C bonds in GO (Fig. 3). The peak shown at 268 nm is due to green reduction of GO. Although, the GO peak absent in the RGO spectrum due to rebuilding of graphitic p-electronic conjugated network structure in RGO after reduction.

Fig. 3. UV–visible spectra of GO and RGO.

FT-IR spectra of GO and RGO (Fig. 4). The existence of oxygen functional groups confirm the

13 Vidya (2019) Vol : 14(2) Vasava et al successful exfoliation of GO. The typical peaks of GO were observed at 1650 cm-1 (C=O stretching) and at 1040 cm-1 (C–O stretching vibration), with a broad peak at 3270 cm-1 (O–H stretching vibrations). The disappearance of peak at 1040 cm-1in RGO spectra shows a decrease of the C– O stretching vibrations. A broad peak at 3270 cm-1 is compressed in the RGO due to removal of hydroxyl groups. This observation supports the reduction of RGO.

Fig. 4. FT-IR spectra of GO and RGO

Dye removal property of RGO was explored with methylene blue dye without interaction to either sunlight or UV using of 20 mg of RGO (Fig. 5). It proposes comprehensive elimination of the dyes in a time span of 50 min.

Fig. 5. Removal of Methyleneblue in the presence of RGO

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Conclusion

X-ray diffraction, SEM imaging, FT-IR and UV-vis spectroscopic techniques clearly reveal the efficient reduction of few layered graphene from GO using green tea extracts. The synthesized RGO was shown to possess suggestively good dye removal properties. The simple and effective method of reduction makes it possibly attractive for the low-cost synthesis of graphene and related materials.

Acknowledgement

The authors are thankful to the Department of Chemistry, Gujarat University, Ahmedabad for providing the essential facilities. The authors are also grateful to MRC-Lab MNIT, Jaipur for XRD and Department of Materials Science, SP University for SEM Analysis. UGC-Info net and INFLIBNET Gujarat University are acknowledged for providing the resource facilities.

References

Abdullah, M. F., Zakaria, R., &Zein, S. H. S. (2014). Green tea polyphenol–reduced graphene oxide: derivatisation, reduction efficiency, reduction mechanism and cytotoxicity. Rsc Advances, 4(65), 34510-34518.

Geim, A. K., &Novoselov, K. S. (2010).The rise of graphene. In Nanoscience and Technology: ACollection of Reviews from Nature Journals (pp. 11-19).

Hummers Jr, W. S., &Offeman, R. E. (1958).Preparation of graphitic oxide. Journal of the american chemical society, 80(6), 1339-1339.

Jin, X., Li, N., Weng, X., Li, C., & Chen, Z. (2018).Green reduction of graphene oxide using eucalyptus leaf extract and its application to remove dye. Chemosphere, 208, 417-424.

Suresh, D., Kumar, M. P., Nagabhushana, H., & Sharma, S. C. (2015). Cinnamon supported facile green reduction of graphene oxide, its dye elimination and antioxidant activities. Materials Letters, 151, 93-95.

Wang, Y., Shi, Z., & Yin, J. (2011).Facile synthesis of soluble graphene via a green reduction of

15 Vidya (2019) Vol : 14(2) Vasava et al graphene oxide in tea solution and its biocomposites. ACS applied materials & interfaces, 3(4), 1127-1133.

Weng, X., Wu, J., Ma, L., Owens, G., & Chen, Z. (2019). Impact of synthesis conditions on Pb

(II) removal efficiency from aqueous solution by green tea extract reduced graphene oxide. Chemical Engineering Journal, 359, 976-981.

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Review Article

ISSN: 2321-1520

A Review on Plant Tissue Culture of Coleus Forskohlii and Its Gene Expression

Sonal kunwar Chouhan, Tisha Patel & Meenu Saraf*

Department of Microbiology and Biotechnology, School of Sciences, Gujarat University, Ahmedabad-380009, India E-mail: [email protected]

*Corresponding Author

Received Date: 6-6-2019

Published Date:15-9-2019

Abstract

Coleus forskohlii Briq.is a perennial herb, belonging to the family Lamiaceae. The tuberous roots of Coleus forskohliia rich source of forskolin (coleonol). It is an endangered plant so it is necessary for the Plant Tissue Culture of Coleus forskohlii. It is a most important medicinal as well as aromatic values. Coleus forskohlii has been successfully micro propagated under in vitro conditions. Aromatic Coleus acts as a natural source of drug for hypertension, cardiovascular health, obesity, glaucoma and many other diseases without any side effects. Worldwide medicinal plant are being investigated because product safe, cheaper, and efficacious in overall.

The pharmacologica and biochemical investigations established that forskolin possess sesmultifaceted biological activities. It is valuable addition in medicinal plants bio diversity. The

demand of this plant is increasing day by day multiplication of this plant is necessary. The herb contains an active ingredient called Forskolin. Forskolin being the major chemical constituent of

17 Vidya (2019) Vol : 14(2) Saraf et al the tuber, herbal preparations of it act on various multiple pharmacologic mechanisms. From gene expression we identified several reference genes that are suitable for qRT-PCR data normalization.

Keywords: Coleus forskohlii, ornamental plant, forskolin, plant tissue culture and gene expression.

Introduction Plant tissue culture is collection of techniques used to maintain or grow plant cells tissue or organs under sterile conditions on a nutrient culture medium of known composition. Medicinal plants are of great significance to the health of individuals and communities. Coleus is an aromatic perennial, with an erect stem and tuber like roots, reaching 60 cm. Coleus is a member of the mint family and native to India, and grows in the subtropical temperate climates of India, Nepal, Thailand and Sri Lanka. The name Coleus is derived from the Greek word Koleos, which means sheath around the style. There are more than 500 varieties of Coleus in cultivation all over the world. Coleus is commonly known as Pashanbhedi (Sanskrit), Patharchur (Hindi), Makandiberuor Manganiberu (Kannada) and is grown throughout the country. Medicinal plants like Coleus have curative properties due to the presence of various complex chemical substances of different chemical nature, which are found as secondary plant metabolites in one or more parts of theseplants. Medicinal plants are important for pharmacological research and drug development, not only when constituents are used directly as therapeutic agents, but also as starting materials for the synthesis of drugs or as models forpharmacologically active compounds (Mukherjee, 200 Complete plants of Coleus froskohliiwas developed in MS medium containing 2.0 mg/l BAP through direct multiplication at the rate of 15-20 shoots per explant. The survival rate of full grown plants is 100 % and rooting occurred in all plants(Kaul et al., 2015). Best shoot formation was reached from the callus using 0.5 mg/l BAP and 0.1 mg/l NAA after 2 weeks of incubation. Green, light green, cremish, brown colour callus was stimulated with (2,4-D) auxin alone and in combination with (BAP) cytokinines ( Atulkar et al., 2012). Transcriptome is the full range of m-RNA, molecules expressed by an organism. Forskolin, a complex labdanediterpendoid found in the root of Coleus forskohlii(Lamiaceae), has received attention for its broad range of pharmacological activities. IriniPateraki (2014) detected Forskolin in the root cork of Coleus forskohliiin a specialized cell type containing characteristic structure

18 Vidya (2019) Vol : 14(2) Saraf et al with histochemical properties consistent with oil bodies. Organelle purification and chemical analysis confirmed the localization of Forskolin and of its simplest diterpene precursor backbone (13R) manoyl oxide, to the oil bodies. The labdanediterpen backbone is typically synthesized by two successive reactins catalyzed by two distinct classes of diterpenesynthases. IriniPateraki et al.(2014) recently described the identification of small gene family of diterpenesynthase candidates (CfTPSs) in Coleus forskohlii.

Geographical Distribution

The Coleus group of plant grows in tropical situations and in warm temperature climate zone on mountains of India, Nepal, Burma, Srilanka, Thailand and Africa. It comes up well on the sun exposed dry hill slopes from 300 m to 1800 m altitude. A well-drained medium fertile soil is suitable for its cultivation.

Botanical Description

Coleus forskohliiis perennial plant that grows to about 45-60 cm tall. It has four angled stems that are branched and nodes are often hairy. Leaves are 7.5 to 12.5 cm in length and 3-5 cm in width, usually pubescent, narrowed in to petioles. Inflorescence is raceme, 15-30 cm in length; flowers are stout, 2 to 2.5 cm in size, usually perfect and calyx hairy inside. Upper lip of calyx is broadly ovate. The ovary is four parted and stigma is two lobed and flower is cross pollinated by wind and insect. The root is typically golden brown, thick, fibrous and radially spreading. Roots are tuberous fasciculate 20 cm long 0.5 to 2.5 cm in diameter, conical fusiform, straight, orange red within and strongly aromatic. Coleus forskohliiis only species of genus to have fasciculate tuberousroots.

Taxonomical Classification of Coleus Forskohlii

Coleus forskohliiBriq.is a member of mint family, Lamiaceae. It is an indigenous to India and is recorded in Ayuvedic material medica under the Sanskrit name Makandi and Mayani (Shah, 1996).The taxonomic position of C. forskohliiis as follows:

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Kingdom - Plantae Divisio - Magnolophyta Class - Magnoliopsida Order - Lamiales Family - Lamiaceae Genus - Coleus Species - forskohlii Genetic Variation in Coleus

Reddy (1952) reported that Coleus forskohliiis diploid with n = 14. However, Riley and Hoff (1961) from their studies on chromosome numbers in South African dicotyledons reported that C. forskohliiis diploid with basic chromosome number n = 16.Birand Saggoo (1982 and 1985) reported that Central Indian collections have basic number of n = 17, while South Indian collections have n =15 and concluded that variability in base number of various members of the family could be due to aneuploidy at generic level which ultimately leads to morphological variations. Shah (1989) reported that populations from different ecogeographic areas vary greatly in their morphology.

Phytochemistry

The tuberous root extracts of C. forskohliicontain minor diterpenoidsviz.,deactylforskolin, 9– deoxyforskolin,1,9- deoxyforskolin, 1,9-dideoxy-7-deacetylforskolin in addition to forskolin (7β- acetoxy-8, 13-epoxy-1α, 6β,9α- trihydroxylabd-14-en-11-one) (Ammon and Kemper,1982 and De Souza and Shah, 1988).

Chemical structure of forskolin Adapted from Khan et al., (2012).

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Importance of Coleus Forskohlii

In traditional Ayurvedic system of medicine, Coleus forskohliibriq.has been used for treating heart diseases, abdominal colic, respiratory disorder, insomnia, and convulsions bronchitis, intentional disorder, burning sensation, constipation, epilepsy and angina. When roots mixed with muster oil, the extract is applied to treat eczema and skin infection. Forskolin is also used in the preparation of medicines preventing hair greying and restoring essential oil has potential color. The plant is used for veterinary purpose (De Souza 1993). Essential oil has potential uses in food flavoring industry and can be used as an antimicrobial agent (Chowdhary et al.,1998).

Action of Forskolin

Forskolin being the major chemical constituent of the tuber, herbal preparations of it act on various multiple pharmacologic mechanisms. De Souza (1977) establish that the methanol extracted from the root tuber is helpful in lowering blood pressure andpositive inotropic activities in animal models. The principle mechanism by which forskolin exerts its hypertensive activity is by stimulation of adenylatecyclase and thereby increasing cellular concentrations of the second messenger cyclic AMP (cAMP) (Seamon et al.,1981). Forskolin directly stimulates almost all hormone sensitive adenylatecyclases in intact cells, tissues and even solubilized preparation of adenylatecyclase (Metzger and Lindner, 1981). The unique feature of this activation is that the site of action for forskolin is the catalytic subunit of the enzyme or a closely associated protein (Seamon and Daly, 1981). Of the 9 types of adenylatecyclase in humans, forskolin can activate all except type IX, which is found in spermatozoa (Iwatsuboet al., 2003). Stimulation of adenylatecyclase is thought to be the mechanism by which forskolin relaxes a variety of smooth muscles. This action of forskolinproved the potential use of the molecule, not only as an invaluable research tool for understanding cyclic – AMP dependent physiological processes, but also as a potential therapeutic agent for diseases like cardiac insufficiency, hypertension, glaucoma, thrombosis, asthma and metastatic condition (Seamon, 1984).

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Adapted from Soni et al., 2012

Use of forskolin

Coleus forskohliiis a herb that has been used before this ancient times in Ayurvedic and Hindu traditional medicine. The roots has been used to treat colic, heart failure, congestive, convulsions, eczema, hypertension, painful urination, insomnia and respiratory disorders (Khan et al., 2012). Uses of Coleus also produce therapeutic benefit in angina, psoriasis, asthma and prevention of cancermetastases.

Cosmetic use

Forskolin is extracted from Coleus froskohliiplant. It is increase isoform selectivity via acting on adenylyl cyclase to enhance the intracellular levels of AMP (cyclic adenosine monophosphate) (Khan et al., 2012). Forskolin in combining with hydroxycitric acid (HCA) can be apply for body shape and fat management (Gupta, 2004). The mechanism of rising and maintaining skimpy body mass is link to the availability of cyclic AMP. Aside facilitating hormonal action, cyclic AMP may manipulate thermo genic response of the body to food, rising the utilization of body and enhance the metabolic rate of the body (Khan et al.,2012).

For skin problems

Coleus has antimicrobial properties. Its oil is useful in topical preparations. Coleus oil is an

22 Vidya (2019) Vol : 14(2) Saraf et al especially effective against propionic bacterium acnes, the microorganism responsible for acne, and it has been found effective against other microorganism known to be responsible for skin infection and eruptions (Khan et al., 2012). This oil suppress the growth of skin pathogens. It has also been found active against yeast culture (Majeed&Prakash, 2007). Antioxidant activity Plant of Coleus froskohliiextract express higher antioxidant activity and high amount of polyphenols in comparison to other Coleus species (Khan et al., 2012). Leaf extract of this plant showed importantly high amount of total polyphenols, flavones and flavonols and high antioxidant activity. Rasineni et al. (2008) intimate that Coleus can be used as an important source of phenolic compound. Perfumery uses Coleus is a rich source of diterpenoids with different oxygen patterns. Over forty compounds consisting to four different classes of aroma compounds have been isolated from oils found from different indigenous genotypes of Coleus forskohli. These include monoterpenoids, diterpenoids, sesquiterpenes alcohol and sesquiterpenes. The presence of 3-decanone, bornyl acetate and g- eudesmol compounds were describe in their experimental studies (Majeed&Prakash, 2007). Miscellaneous uses Coleus also well work in treating cancer, asthma, cardiopathy, congestive heart failure, cramp, convulsions, depression, dermatosis, dyspepsia, dysuria, glaucoma, eczema, high blood pressure, hypothyroidism, infertility, insomnia, ischemia, myocardosis, obesity psoriasis, respirosis, water retention and thrombosis (Duke et al., 2002). Heart disorder Forskolin has a positive inotropic action on cardiac tissue via increased cAMP levels. Forskolin lowered normal or elevated blood pressure in different animal species through a vasodilatory effect and it had a positive inotropic action on the heart muscle (De Souza et al.,1983; Dubey etal.,1981).

Glaucoma

The topical utilization of forskolin lowered the intraocular pressure in rabbits, monkeys and healthy

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human volunteers and it was associated with a reduction in aqueous inflow and no change in outflow facility indicating the potential of forskolin as a therapeutic agent in the treatment of glaucoma. It also showed no influence on humans in reducing aqueous flow when applied topically to the eye (Brubaker et al., 1987). Asthma

Forskolin was studied as bronchodilator for its potential use in the treatment of asthma (Bruka, 1986). In human basophils and mast cells, forskolin blocked the release of histamine and leukotriene C-4 (Marone et al., 1987). A study involving human revealed that inhaled forskolin powder formulations were capable of causing bronchodilatio in asthma patients (Bauer et al., 1993).

Antithrombotic effect

Forskolin inhibits platelet aggregation through adenylatecyclase stimulation, augmenting the effects of prostaglandins (Siegl et al., 1982; Adnot et al., 1982).

Anti-obesity

The antiobesity effects of C. forskohliiwere investigated in ovariectomized rats (Han et al., 2005) and the administration of C. forskohliiextracts reduced body weight, food intake and fat

accumulation in those rats suggesting that C. forskohliimay be useful in the treatment of obesity.

Other uses

In addition to its cAMP stimulating activity, forskolin inhibits the binding of platelet-activating factor (PAF), independently of cAMP formation (Wong, 1993). Forskolin also appears to have an effect on several membrane transport proteins and inhibits glucose transport in erythrocytes, adipocytes, platelets and othercells (Mills et al., 1984). The safety of C. forskohliiand forskolin has not been fully evaluated. It should be avoided in people with ulcers, because it may increase stomach acid levels (Seamonetal., 1981).

Plant Tissue Culture of Coleus Forskohlii Plant tissue culture refers to in vitro techniques used to cultivate the plants, seeds and various

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parts of the plant like embryo, tissues, protoplast etc. Plant cell has the ability of totipotency that can differentiate and regenerate from meristematic cell to whole plant. Coleus forskohliiis an endangered plant so it is necessary for growth and development of its by plant tissue culture. Develop complete plant free from microbes by the plant tissue culture technique. High frequentation Shoot generation of Coleus forskohlii In plant tissue culture developed complete plants of Coleus forskohliiin MS medium containing 2.0 mg/l BAP through multiplication at the rate of 15-20 shoots per explants (KaulT etal.,2015). Shoot regeneration from callus As callus culture can be obtained as rapid plant regeneration system which could be used for the somaclonal variationfor producing transgenic plants. Developed plant of Coleus forskohliifrom the leaf explants of callus. For callus formation using the growth hormones 2,4-D (1.0 mg/l) alone and in combination with BAP (0.5 mg/l) induced green colour,browncolour, greenfragile, and light green colourare formed. Best shoot induction are achieved from the callus using BAP 0.5 mg/l and NAA 0.1mg/l after two week of inoculation. Where highest number of shoot induced and maximum frequency was regenerated. Pratibhadube (2011) was developed the plant of Coleus forskohlii from the adventitious shoots in MS medium supplemented with 1.5 mg/lBAP. Callus formation of Coleus forskohlii Malathy and J.pai (1999) have obtained the callus using the hormone of cytokinin and auxin. E sreedevi (2013) was developed plant from the callus on MS medium containing in BAP (2mg/l) and NAA (1mg/l). The highest rate of shoot multiplication was achieved and more than 200 shoots were produced from the callus clump. Callus induction and proliferation system are known to be very useful for the study of bio-synthesis of natural product of Coleus forskohlii(Sen et. al.,1992; Mukherjee et. al.,1996; Reddy et.al.,2001). Gene expression Stable reference genes are necessary to normalize the gene expression data (Shin-young hong et al (2008). Reference genes are particularly useful when stress responsive gene are analyzed in order to produces transgenic plants that exhibit enhanced stress resistance. Genes encoding tubulins, actin or elongation factors are widely used in gene expression studies. We will use three genes

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Actin7 (ACT7), Tubulin alpha 6 (TUA6), and Ubiquitin18 (UBC18) for gene expression studies. UBC18 and ACT7 genes are the most stable genes that might be widely used for multiple purposes. The function of ACT7 genes encode one of the six distinct and ancient subclasses of actin protein in the complex Arabidopsis actin gene family. ACT7 actin genes and investigated its tissue specific expression and regulation. The highest level of ACT7 mRNA are found in rapidly expanding vegetative organs.TheACT7 mRNA levels varied by 128- fold among different tissue and organs.TheACT7 promoter may contain a high density of hormone response elements (J. McDowell et al,1996). TUB6 genes the expression patterns of the affected tubulin genes and the phenotypes of the lefty double mutant to address increasing effects of microtubule defects on cell morphogenesis. Tubulin genes were expressed ubiquitously in examined tissue and cell types in Arabidopsis (Tatsuya Abe, et al 2004). The UBC18 gene encoding an ubiquitin-conjugating enzyme exhibits the most stable expression in different plant tissue or in plants treated with various growth hormones. The ubiquitin-conjugating enzyme UBC18 function redundantly to control an early step of pharyngeal morphogenesis. The regulation of protein stability through ubiquitin-mediated proteolysis has been show to play a critical role in a wide range of cellular and developmental processes. In Brachypodiumdistachyonthe expression of the polyubiquitingenes are most stable in different plant tissue and growth hormone treated plant sample, the expression of the S- adenosymethionine decarboxylase gene (Sam DC) ranked are most stable in plant grown under various environmental stresses (Shin-Young Hong et al.2008).

Conclusion

The present evidence shows that Coleus forskohliiis the only known natural source of the forskolin (deterpenoid). Moreover still, there is paucity for the mechanism of other bioactive principles present in the herb except forskolin. Further researches in view of applicability of forskolin for treating human ailments without any side effects. It is an ornamental plant with many herbal component and herbal properties. It is a valuable addition in medicinal plants biodiversity. The expression of growth regulatory genes were seen to be upregulated in Coleus treated explants and hence it can be observed that Coleus can be propagated by plant tissue culture method in a short duration and conserved environment.

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Reference Abe, T., Thitamadee, S., & Hashimoto, T. (2004).Microtubule defects and cell morphogenesis in the lefty1lefty2 tubulin mutant of Arabidopsis thaliana.Plant and Cell Physiology, 45(2), 211- 220. Adnot, S., Desmier, M., Ferry, N., Hanoune, J., &Sevenet, T. (1982). Forskolin (a powerful inhibitor of human platelet aggregation).Biochemical pharmacology, 31(24), 4071-4074. Bauer, K., Dietersdorfer, F., Sertl, K., Kaik, B., &Kaik, G. (1993).Pharmacodynamic effects of inhaled dry powder formulations of fenoterol and colforsin in asthma.Clinical Pharmacology & Therapeutics, 53(1), 76-83. Biswas, T. K., & Mukherjee, B. (2003). Plant medicines of Indian origin for wound healing activity: a review. The international journal of lower extremity wounds, 2(1), 25-39. Brubaker, R. F., Carlson, K. H., Kullerstrand, L. J., & McLaren, J. W. (1987).Topical forskolin (colforsin) and aqueous flow in humans.Archives of Ophthalmology, 105(5), 637-641.. Bruka, J. F. (1986). Forskoli: Its chemical biological and medical potential. In Proc. Int. Symp.Forskolin, Hoechst India Ltd., Bombay (p. 1986). De Souza, N. J., Dohadwalla, A. N., &Reden, Ü. (1983). Forskolin: a labdanediterpenoid with antihypertensive, positive inotropic, platelet aggregation inhibitory, and adenylatecyclase activating properties. Medicinal research reviews, 3(2), 201-219 Dube, P., Gangopadhyay, M., Dewanjee, S., & Ali, M. N. (2011). Establishment of a rapid multiplication protocol of Coleus forskohliiBriq. and in vitro conservation by reduced growth. Duke, J. A. (2002). Handbook of medicinal herbs.CRC press. Han, L. K., Morimoto, C., Yu, R. H., & Okuda, H. (2005).Effects of Coleus forskohliion fat storage in ovariectomized rats.Yakugakuzasshi: Journal of the Pharmaceutical Society of Japan, 125(5), 449-453. Hedin, L., &Rosberg, S. (1983). Forskolin effects on the cAMP system and steroidogenesis in the immature rat ovary. Molecular and cellular endocrinology, 33(1), 69-80.

Hong, S. Y., Seo, P. J., Yang, M. S., Xiang, F., & Park, C. M. (2008).Exploring valid

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reference genes for gene expression studies in Brachypodiumdistachyonby real-time PCR.BMC plant biology, 8(1), 112.

Kaul, T., Malik, M. A., Yaqoob, U., & Mehta, J. (2015).High Frequency and Rapid in vitro Plant Regeneration ofColeus forskohliiBriq.Med Aromat Plants, 4(193), 2167-0412.

Kavitha, C., Rajamani, K., &Vadivel, E. (2010).Coleus forskohliiA comprehensive review on morphology, phytochemistry and pharmacological aspects. Journal of Medicinal Plants Research, 4(4),278-285.

Khan, B. A., Akhtar, N., Anwar, M., Mahmood, T., Khan, H., Hussain, I., & Khan, K. A. (2012). Botanical description of Coleus forskohlii: a review. Journal of Medicinal Plants Research, 6(34),4832-4835.

Khatun, S., Chatterjee, N. C., &Cakilcioglu, U. (2011). The Strategies for Production of Forskolinvis-a-vis Protection Against Soil Borne Diseases of the Potential Herb ColeusforskohliiBriq.

Majeed, M., &Prakash, L. (2004).Fighting acne and more Effective natural approaches to skin care. Cosmetics & Toiletries Manufacture Worldwide, 215-219.

Malathy, S., & PAP, J. (1999). Monitoring of forskolin production from roots and callus by HPTLC in Coleus forskohliiBriq. Journal of Spices and Aromatic Crops, 8(2), 153-157.

McDowell, J. M., An, Y. Q., Huang, S., McKinney, E. C., & Meagher, R. B. (1996). The Arabidopsis ACT7 actin gene is expressed in rapidly developing tissues and responds to several external stimuli. Plant physiology, 111(3), 699-711.

Metzger, H., & Lindner, E. (1981). The positive inotropic-acting forskolin, a potent adenylatecyclase activator. Arzneimittel-Forschung, 31(8), 1248-1250..Shah VC(1989).

Pateraki, I., Andersen-Ranberg, J., Hamberger, B., Heskes, A. M., Martens, H. J., Zerbe, P., ...&Hamberger, B. (2014). Manoyl oxide (13R), the biosynthetic precursor of forskolin, is synthesized in specialized root cork cells in Coleus forskohlii. Plant physiology, 164(3), 1222- 1236.

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Plectranthusforskohlii(Wild) Briq.(Syn: Coleus forskohlii)—A compendium on its botany and medicinal uses. Int.J. Res. Plant Sci, 3, 72-80.

Qiu, X., & Fay, D. S. (2006). ARI-1, an RBR family ubiquitin-ligase, functions with UBC-18 to regulate pharyngeal development in C. elegans. Developmental biology, 291(2), 239-252.

Riley, H. P., & Hoff, V. J. (1961). Chromosome studies in some South African dicotyledons. Canadian Journal of Genetics and Cytology, 3(3), 260-271.

Seamon, K., & Daly, J. W. (1981). Activation of adenylatecyclase by the diterpeneforskolin does not require the guanine nucleotide regulatory protein. Journal of Biological Chemistry, 256(19), 9799-9801

Soni, H., &Singhai, A. K. (2012). Recent updates on the genus coleus: A review. Asian J Pharm Clin Res, 5(1), 12- 17.. 8.

Tharp, M. D., Kagey-Sobotka, A., Fox, C. C., Marone, G., Lichtenstein, L. M., & Sullivan, T. J. (1987). Functional heterogeneity of human mast cells from different anatomic sites: in vitro responses to morphine sulfate. Journal of allergy and clinical immunology, 79(4), 646-653

Wong, S., Mok, W., Phaneuf, S., Katz, S., &Salari, H. (1993). Forskolin inhibits platelet- activating factor binding to platelet receptors independently of adenylyl cyclase activation. European Journal of Pharmacology: Molecular Pharmacology, 245(1), 55-61

xSeamon, K. B., Padgett, W., & Daly, J. W. (1981). Forskolin: unique diterpene activator of adenylatecyclase in membranes and in intact cells. Proceedings of the National Academy of Sciences, 78(6), 3363-3367.

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Research Paper

ISSN: 2321-1520

Using Agricultural Residues for Biomass Briquetting and pelletising asa Source of Renewable Energy to replace fossil energy by biomass co-firing to mitigate climate change and address farmer’s poverty by enhancing farmer’s income

Vineshsingh Rore* , Himanshu Pandya, A. U. Mankad

Department of Botany, Bioinformatics and Climate change Impacts Management University School of Sciences,Gujarat University

*Corresponding Author

Received Date: 30-7-2019

Published Date: 15-9-2019

Abstract

Achievement of long term climate change mitigation objectives will ultimately require a transition from fossil fuel energy to renewable energy. Every year millions of tons of agricultural wastes are generated which are either destroyed by left to decay on their own in fields generating methane (CH4) or burnt inefficiently in loose form by carbon dioxide (CO2)) causing air pollution and global warming as both are potential greenhouse gases. The agricultural residues like rice husk, wheat straw, castor shells, ground nutshell and cotton stalks etc. generated in abundance in state of Gujarat which are either left to decay on their own or burnt in fields to clear fields for next sowing season of crops can be utilized as the potential biomass energy sources to be densified into pellets and briquettes for biomass co-firing as well as replacement of fossil energy and fuels .This renewable biomass energy fuel is beneficial for the environment as it conserves natural resources

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as well as generates additional income for farmers whereby instead of spending money on getting rid of these agricultural residues from fields farmers can actually earn handsome amount of money by selling these agricultural residues and farm wastes and also simultaneously contribute to green- house mitigation and climate-change mitigation. In this paper we examine agricultural residues like rice husk, wheat straw,saw dust, castor shells,ground nutshell and cotton stalks etc generated in abundance in state of Gujarat as the potential biomass energy sources to be densified into pellets and briquettes for biomass co-firing as well as replacement of fossil energy and fuels. We discuss the various advantages, factors that affecting the biomass briquetting and comparison between coal and biomass briquetting. The de- tails of the study were highlighted in this paper. Keywords- Biomass, Briquetting, Pelleting, Potential, Process, Technologies Introduction Concerns regarding the potential global environmental impacts of fossil fuels used in power gen- eration and other energy supplies are increasing worldwide. The primary driver of climate change is the emission of greenhouse gases, including carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O) that arise from a number of human activities, the primary contributors being defores- tation and the burning of fossil fuels for energy generation. Achievement of long term climate change mitigation objectives will ultimately require a transition from fossil fuel energy to renewable and sustainable energy. A number of techniques and methods have been proposed for reducing gaseous emissions of NOx ,SOx and CO2 from fossil fuel combustion and for reducing costs associated with these mitigation techniques. Some of these control methods are, however, expen- sive and increase the overall production costs. Biomass Co-Firing : A Potential Option To A Clean Future One of the key areas of environmental concern globally is power generation in coal power plants. Coal fired power plants account for 41% of global electricity, and the importance of coal to electricity generation worldwide is set to continue with coal fuelling 44% of global electricity in 2030. There is an increasing interest worldwide in generation of energy from biomass, as a carbon neutral alternative to coal. Among the quickest and most cost-effective alternatives for large elec- tric utilities producers is the technique of “Biomass cofiring”

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Biomass cofiring refers to the simultaneous combustion of a biomass fuel and a base fuel to pro- duce energy, usually electrical power. “Biomass co-firing” in coal based power plants involves replacing a certain percentage of coal the biomass briquettes/pellets to reduce GHG emissions at source. This technology is emerging as a sustainable model for mitigating environmental degrada- tion due to carbon emissions from large coal based power plants.

The technology has successfully moved from engineering studies to parametric tests to long term demonstrations. As it gains acceptance worldwide, co-firing technology will address many envi- ronmental issues facing the world today.

Worldwide, about 40% of electricity is produced using coal. If only 5% of coal energy is replaced by biomass in all coal-fired power plants, this would result in CO2 emission reduction of around 300 Mton CO2/ year.

Co-Firing With Biomass Briquettes/Pellets

Co-firing is the simultaneous combustion of a supplementary fuel with a base fuel. The most com- mon base fuel is coal. Biomass briquettes/pellets are a carbon neutral, renewable fuel source produced using farm residues, left over forest residues and other natural organic stuff, as well as through farming of dedicated energy crops and plants on marginal or wasteland.

Co-firing with biomass briquettes/pellets incorporates environmental, socio-economic and larger strategic advantages including.

• Lower capital costs for power generation.

• Reduced coal usage without major technology and infrastructural changes.

• Highest electrical conversion efficiencies amongst all biomass power technologies.

• Proven model world wide in installations for most fuel combinations and boiler types.

• A clean, renewable fuel for power plants with reduction in GHG emissions and ash content.

• People friendly alternative that improves hygiene, maintains cleanliness and lowers dust gen- eration.

• Ease of compliance with emission norms as compared to solar and wind technologies.

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• Greater employment and income generation opportunities to support rural economies.

• Encourages the formation of a biomass commodity market.

Co-firing with biomass briquettes/pellets is, therefore, a long term, sustainable alternative that has the potential to meet the global climate challenge, as well as generate grassroots growth and development for nations.

About Biomass Briquettes/Pellets

Half a kilo of dry plant tissue can produce as much as 1890 kcal of heat which is equivalent to the heat available from a quarter of kilogram of coal.

Biomass is a renewable energy resource derived from numerous sources, such as left over farm residues, left over forest residues and other natural organic stuff, as well as farming of dedicated energy crops and plants. Biomass is the most important fuel worldwide after coal, oil and natural gas.

Biomass is abundantly available on earth in the form of agricultural residues that lie unutilized and can be effectively used for power generation.

Biomass Briquettes/Pellets /Pellets

Biomass briquette is a fuel material that is uniform insize, shape, moisture, density and energy content, produced by drying and compacting different types of biomass. During the production process, biomass is dried in the most efficient way in order to ensure maximum heat generation when it is burned in the boiler.

Biomass Briquettes/Pellets/Pellets Are Carbon Neutral

From a chemical perspective, biomass briquettes/pellets contain carbon in simple or more com- plex combinations with other chemical elements - oxygen, hydrogen, etc. Carbon is the base element, which can be burned (by oxidizing it) to produce heat energy, which can thereafter be converted into other types of energy -steam, electricity, etc.

From an ecological point of view, biomass briquettes/pellets is an ideal fuel material because it is considered carbon neutral. Using biomass briquettes/pellets/pellets as an energy source creates a

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'closed carbon cycle'. As a biomass energy source grows, CO2 is absorbed from the atmosphere and when it is burned the same amount of CO2 stored by the biomass is released ensuring that the natural carbon cycle balance is maintained. This makes biomass briquettes/pellets “carbon neu- tral.”

As opposed to this, fossil fuels like coal are extracted from below the earth, releasing CO2 trapped beneath the soil, into the atmosphere. As this process continues, there is a CO2 overload in the atmosphere, which disturbs the natural carbon cycle, and is one of the culprits behind climate change. Fossil fuels are, therefore, considered to be “carbon negative.”

Carbon dioxide emissions from the burning of fossil fuels currently account for about 65 per cent of the extra carbon dioxide in our atmosphere. Carbon from biomass is recycled within one year whereas carbon from fossil fuels such as coal takes 9000 to 12000 years for recycling.

Benefits of Using Biomass Briquettes/Pellets

Environmental Benefits

Biomass briquettes/pellets V/s Coal & Lignite

Coal is a non renewable source of energy.Burning coal emits harmful gases and waste such as carbon dioxide, sulphur dioxide, nitrogen oxides, sulphuric acids, arsenic and ash, and has led to major environmental impacts like acid rain in some regions. Coal excavation requires a significant amount of energy and it is generally transported from remote locations through long distances, leading to higher carbon emissions.

Lignite causes problems in transportation and storage, due to its high moisture and ash content, and is susceptible to spontaneous combustion. It is also difficult to crush, pulverize, and combust. It has a lower heating value, which means that more fuel must be handled to produce a given amount of power. The high inherent moisture content of lignite decreases boiler efficiency. Its ash characteristics require more attention to soot blowing and boiler operation to maintain high avail- ability and reliability.

Variations in quality parameters of lignite /coal like ash content, moisture, GCV result in efficiency losses and variations in steam pressure. High SPM content is also an area of concern, especially in

34 Vidya (2019) Vol : 14(2) Pandya et al the case of lignite, and adding to this is the pressure to abide by very strict laws of pollution control boards

Biomass briquettes/pellets manufacturing and transportation leads to lower carbon emissions as compared to fossil fuels, as biomass is abundantly available on site and can be easily collected without significant use of energy. Biomass, after collection, is processed in a vicinity of 50-100 kms from the site of cultivation/harvest, which further reduces the carbon emissions from process- ing. Biomass briquettes/pellets are a carbon neutral, environment friendly fuel that provide a com- fortable, healthy working environment to the manpower involved in collection, processing, manu- facturing and usage.

Carbon Emission Reduction Potential

Biomass briquettes/pellets co-firing reduces/displaces fossil fuel use in large coal fired power plants with higher CO2 emission levels. Recognizing that about 50% of tree mass (on a dry basis) is carbon, tree energy crops represent a significant tool for carbon management-having an additional sequestration component which solar/wind energy do not have.

Reduced GHG Emissions

Co-firing biomass briquettes/pellets with coal reduces concentration of GHG emissions by using waste residues, which would otherwise cause carbon dioxide emissions through open burning, or methane emissions through decay.

Sustainable use of biomass briquettes/pellets also addresses the threats associated with global warming, because they are carbon-neutral and reduce SOx emissions to significantly lower levels as compared to conventional fuels.

Economic Benefits

Rural Income & Employment Generation

The use of biomass briquettes/pellets as an energy source generates at least 20 times more local employment within the national economy than any other form of energy per unit. This is because a large amount of unskilled labor is engaged in growing, harvesting, processing, transporting and trading the fuels, which generates off-farm income for rural populations, either regularly or off-

35 Vidya (2019) Vol : 14(2) Pandya et al season. Thus, short term and long term employment is created through project activities associ- ated with biomass energy.

There is also an opportunity for revenue generation for suppliers of agro residues viz., small-scale industries and farmers, which enhances development of the region and uplifts the local economy. It also fosters a sense of self reliance in local and rural communities and empowers them to a great extent.

Money Velocity within the Region The most compelling principle of using biomass briquettes/ pellets in co-firing is that, since biomass is cultivated regionally, there is never a monetary drain on a town, city or country. Regional waste problems are addressed and supply is tailored to local needs.

Energy Efficiency V/s Wind & Solar

The average PLF for biomass power projects is much higher as compared to solar and wind, resulting in a significantly higher efficiency of power generation This means that for 1MW power to be generated, biomass stand alone or existing coal fired power plants (where co-firing needs to be done) need to have an installed capacity of 1MW, as the PLF is 0.8. In wind and solar power, the installed capacity needs to be as high as 4 MW or 5 MW as the PLF is very low, 0.2 and 0.15, respectively.

Strategic Benefits

Investment & Benefits within Nation

Biomass based power generation requires comparatively less investment in technology than solar and wind energy, which are highly capital-intensive technologies.

This allows for local industry investments, and larger benefits to the nation in the form of enhanced income and employment generation opportunities.

Increased Energy Security

Local availability and reliability of supply is one of the key advantages of biomass briquettes/ pellets/pellets. Modern applications have demonstrated that biomass briquettes/pellets energy can

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be competitive for large-scale industrial applications. For fuel importing countries, the use of local biomass can save substantial amounts of foreign exchange and reduces dependence on other countries for conventional fuels.

Co-Firing In Existing Thermal Power Plant

Co-firing can be done in existing power plants with little or no modifications, allowing for com- paratively inexpensive technologies and resulting into rapid reductions in greenhouse gases and local pollutants like SOx ,NOx and SPM.

Besides their potentially harmful effect on the environment, which is well documented, fossil fuels are also a scare resource, and the world is constantly looking for less harmful, more cost effective, and abundantly available renewable energy alternatives, that can replace fossil fuels in energy generation.

Biomass is a widely available resource that can be a powerful substitute for fossil fuels.

Co-firing biomass briquettes/pellets and coal is a win-win combination that offers several key advantages.

Co-firing can be done in existing power plants with little or no modification, allowing for compara- tively inexpensive and rapid reductions in greenhouse gases.

It takes advantage of the high efficiencies obtainable in coal-fired power plants.

It also maintains fuel diversity ,which reduces the need for a constant supply of biomass briquettes/ pellets/pellets that would be required in a stand alone biomass power plant.

Co-Firing Helps Meet Renewable Power Purchase Obligation (Rppo) Norms

Different strategies are promoting renewable energy sources for power generation globally. Biom- ass is a strong contender for renewable energy across the world.

In India too, power sector reforms are underway and the Indian government is promoting renew- able energy as a tool to control energy-related GHG emissions.

The Renewable Power Purchase Obligation (RPPO) clearly sets a mandate for including the contribution of renewables to electricity generation.

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As co-firing needs less capital investment and less technological modifications, it emerges as a credible renewable technology and is the quickest and most cost effective option to help the country meet its RPPO norms.

Co-Firing With Biomass Briquettes/ Biopellets Vis-À-Vis Loose Biomass

Instead of burning the loose biomass fuel directly, it is more practical to compress it into pellets (compressing them through a process to form blocks of uniform shapes) and thereby improve its utility and convenience of use.

Biomass briquettes/pellets can also be used directly as fuel instead of coal in the boilers.

Biomass Briquettes/Pellets V/S Loose Biomass

Co-firing biomass in a dense briquetted form, known as biomass briquettes/pellets, offers a lot of advantages such as ease in handling, convenience in use, proper combustion and efficient trans- portation.

Biomass briquettes/pellets are uniform in size, shape, moisture, density and energy content. During its production, biomass is dried in the most efficient way in order to ensure maximum heat genera- tion when it is burned in the boiler. As opposed to this, when burning wet biomass, additional energy is consumed during the burning process to dry the biomass, thereby increasing the total fuel material consumption and decreasing heating efficiency.

Due to the high density of biomass briquettes/pellets, they can be handled more easily and predicatably in large-scale applications. They also allow for a smaller and simpler conveying sys- tem that reduces costs and supports free flow of the biomass fuel through the conveyor system.

There is no technology upgradation required in terms of capacity enhancement for biomass bri- quettes/pellets, whereas with loose biomass, it is important to consider the possibility of deteriora- tion of capacity due to use of less densified fuel, which may require additional investments in capacity enhancement.

Biomass briquettes/pellets reduce the actual fuel consumption because of proper combustion in the furnace area, thereby increasing the efficiency of the system. It is convenient for continuous

38 Vidya (2019) Vol : 14(2) Pandya et al mechanical feeding operation and there is an increase in the life of the boiler.

The high density and uniform shape of biomass briquettes/pellets enables their storage in standard silos. Less storage space is required due to piling of standard sized bags, resulting in savings in storage space and manpower costs, as well as optimal utilization of space. In contrast, loose biomass requires more space due to irregular shaped packaging and piling of bags, leading to increased storage and manpower costs. Biomass briquettes/pellets are also more efficient to transport than loose biomass, because no slack, air and water is being transported.

Conclusion

The use of densified agri wastes for renewable energy for co-firing for power generation and along with fossil fuels for energy needs in Industry would serve as an effective tool of climate change adaptation and mitigation strategy by decreasing carbon-di-oxide and Methane emissions associ- ated with the burning and decay of crop residues and agri-wastes in the agricultural fields after harvesting of crops as well as providing effective renewable energy fuel substitute in the form of biomass briquettes for the energy needs of the country.

It would also contribute to sustainable socio-economic development with social equity as the major project beneficiaries, would be rural population especially in the agricultural dominated regions where agri-waste and crop residue are generated and are available in abundance and the local farming community has a major challenge in disposing them before the next sowing season from their small marginal fields and also would ensure increased access to low cost, clean energy fuel for rural businesses and households as well as industry, institutes, commercial units at costs comparable to the existing fuels being used by them.

It also has capability for ancillary industries development. It would also result in establishment of biomass briquetting and pelleting machine spares and consumables manufacturing units and work- shops for manufacturing Die, Ram, Collates, Collete Flenge, V Belt etc locally. This would be a win-win situation for all the stake holders viz. Farmers, Agri enterprenuers, Rural community and Households. Thus it would result in sustainable socio-economic development in the region and effectiveness as the tool of climate change adaptation and mitigation strategy which is an important aspect of the it leading to its widespread acceptance among the various stake holders.

39 Vidya (2019) Vol : 14(2) Pandya et al

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Research Paper

ISSN: 2321-1520

Phytochemical Screening of Blumeaeiantha Dc. and Sphaeranthusindicus L.

Vibhuti S. Patel*, N.R. Modi

Department of Botany, Bioinformatics & Climate Change Impacts Management, School of Science, Gujarat University, Ahmedabad. E-mail: [email protected]

*Corresponding Author

Received Date: 8-7-2019

Published Date:15-9-2019

Abstract

The aim of the present study was qualitative phytochemical screening of aqueous and methanolic extract in leaves of Blumeaeriantha DC and SpharenthusindicusL.belongs to Asteraceae family. This genus includes various species which are used in preparation of traditional and endemic medicines.Blumeaeriantha is known to show anti-diarrheal, anticancer and anti-diabetic activity while Spharenthusindicuswas found to possess powerful medicinal properties to cure skin infection, liver disease, jaundice, bronchitis. The phytochemical screening of aqueous and methanolic leaf extracts revealed the presence of various secondary metabolites such as alkaloids, phytosterols, phenolic compounds, tannins, flavonoids, terpenoids and saponins.

Keywords: Blumeaeriantha DC, Spharenthusindicus L., phytochemical screening.

Introduction

"A medicinal plant" which contains organic compound can be used for therapeutic purpose in the

49 Vidya (2019) Vol : 14(2) Patel et al

form of drugs. The Medicinal plants have great value which gives a significant physiological action in human body (Edeogaet al, 2005). The medicinal plants have backbone of traditional herbal medicine. More than 400 different plant species are listed for medicine and their medicinal values in Ayurveda, Unani, Homeopathy, Sidhha. Phytochemical means ‘phyto-from Greek - phyto meaning ‘plant’. Generally, plant chemical that are protecting the plant cell from environment risk like stress, pollution, and UV exposure (Mamta s. et al, 2013). Phytochemicals are classified mainly in two groups primary metabolites and secondary metabolites. The Medicinal plants have variety of phytochemicals such as carbohydrate, proteins, amino acids, alcohols, oils, tannins, saponins, glycosides, Phenol and flavonoids.

The Blumeaeriantha Dc and Spharenthusindicus L. belongs to Asteraceae family. Blumeaeriantha is an aromatic, small annual herb, height upto 1 m, camphor like smell, and commonly found in the road side of near forest area. It is commonly known as "Nirmundi" and "Kukronda" (Singh et al.,2011).Sphaeranthusindicus is small aromatic herb, height 15-30 cm, reddish-purple of ovoid heads or solitary glandular peduncles with toothed wings. The plant grows in rich field, dry waste places and cultivated lands in tropical parts (Chatterjee et al.,1997 and Galani et al., 2010)

Material and Method

Collection of plant material

The whole plant was collected from motapondha region of valsad district, Gujarat in the month of January 2018. The collected plant material was identified and authenticated by Dr.Sandip Patel, taxonomist, Dr. A.P.J. Abdul KalamSilvassa College, Gujarat. Taxonomic identification was carried out in the Department of Botany, School of Science, Ahmedabad.

Preparation of Sample

Fresh leaves and stem of Blumeaeriantha andSpharenthusindicuswere washed in water and naturally air dried at room temperature for 10-15 days. Then make fine powdered with the help of mixer grinder and store in air tight bottles.

Preparation of Extraction

Dried samples powered were weighted 10 gm and kept in flask containing 100 ml solvents (Methanol

50 Vidya (2019) Vol : 14(2) Patel et al and distilled water) on shaker at room temperature. After 24 hours the solution was filter using Whatman no 1 filter paper and allowed it to evaporate at the room temperature.

Result

Qualitative analysis

The Qualitative screening of BlumeaerianthaDc. and SphaeranthusindicusL. was performed according to standard protocol by harborne 1998, kokate 1999, Raaman 2000.The present phytochemical constituents are shown in table 1.

Phytochemicals Tests Aqueous Methanol

Blumea Sphaeranthus Blumea Sphaeranthus

Alkaloids Mayer’s test ++ +++ + +

Wagner’s test ++ +++ ++ ++

Dragendroff test ++ +++ + ++

Hager’s test ++ +++ + +

Carbohydrates Molish test _ _ _ _

Fehling test _ _ _ _

Barford test _ _ _ _

Bendict test _ _ _ _

Flavonoids Alkaline reagent +++ ++ +++ ++ test

Lead acetate test ++ ++ +++ ++

Saponin Froth test + + _ _

Foam test ++ ++ _ _

51 Vidya (2019) Vol : 14(2) Patel et al

Terpenoids Salkowski test +++ _ +++ _

Copper acetate test ++ _ +++ _

Phenols Ferric chloride test +++ ++ +++ +++

Tanins Gelatin test +++ ++ ++ ++

Ferric chloride test ++ ++ ++ ++

Glycosides Legal’s test + + _ _

Kellakiliani test + + _ _

Phytosterols Salkaouski’s test ++ + ++ +

Lieberman + + + + burchard’s test

Table 1: Qualitative screening of BlumeaerianthaDc. and SphaeranthusindicusL. (+=present, - = absent)

Conclusion

This present study concluded that the phytochemical of Blumeaeriantha has alkaloids, flavonoids, terpenoids, phenols, tannins, phytosterols are present while saponins and glycosides are present only in aqueous extract, but carbohydrate is absent in both extracts. In aqueous and methanolic extract of Spharenthusindicusalkaloids, flavonoids, phenols, tannins, phytosterols are present whilesaponins and glycosides are present only in aqueous extract, but carbohydrate and terpenoids are absent in both extracts.

References

Chatterjee A, Pakrashi S C (1997), The treatise on Indian medicinal plant, Vol 5, National Institute of Sciences Communication and Information Resources, New Delhi.

Edeoga HO, Okwu DE, Mbaebie BO (2005). Phytochemical constituents of some Nigerian medicinal plants, Afr. J. Biotechnol. 4(7): 685-688.

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Galani VJ, Patel BG, Rana DG, Sphaeranthusindicus L.: A Phytopharmacological Review, International Journal of Ayurveda Research,1,2010,247-253.

Harborne JB (1998). Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. New Delhi: Springer, Rajkamal Electric Press, 3rd edn. p.1-32, 40-227.

Mamta S, Jyoti S, Rajeev N, Dharmendra S and Abhishek G(2013).Phytochemistry of medicinal plant, Journal of Pharmacognosy and phytochemistry ,1(6): 168-182.

Singh UP, Singh AK, Sarathy R P (2011), Effect of Methanolic extracts of Blumeaeriantha DC leaves on protein metabolism and marker enzyme s in streptozotocin induced Hyperglycemicanimals. Int J of pharmaSci; Vol 4(1):235-238.

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Research Paper

ISSN: 2321-1520

Bioactive Natural Compounds from Cactus Euphorbia Caducifolia

Maitry Acharya1, Maitry Mehta1, Zalak Rathod*, Meenu Saraf2

Department of Microbiology and Biotechnology,School of Sciences, Gujarat University, Ahmedabad 380009 E-mail:[email protected]*

*Corresponding Author

Received Date: 12-6-2019

Published Date:15-9-2019

Abstract

In recent years, bioactive compounds are in high demand in the pharmaceuticals and naturopathy, due to their health benefits to human and plants. The bioactive of bioactive compounds. Bioactive compounds like alkaloids, steroids, compounds are extra-nutritional constituents present in small quantities in lipid rich foods and plant products. Endophytes are organisms that live inside healthy tissue of plant. Majority of endophytic bacteria showed beneficial effects like enhancement of biological nitrogen fixation, production of phytohormons, solubilization of phosphate etc. Endophytic bacteria, fungi and actinomycetes plays significant role in production flavonoids, quinols, phenols. Cacti have attracted special interest due to their particular traits, such as a spinosebody that stores water and photosynthesize with CAM metabolism. Such traits allow them to live in extreme environments and tolerate water stress.

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Introduction

Endophytesan overview

Endophytes are group of microorganisms having the ability to enter inside the plant hosts colonizing the intercellular spaces and also the xylem vessels. Almost all plants species have been found to harbour endophytic bacteria or fungi (Sturz and Nowark 2000). Moreover, the colonization of endophytes in algae (Smith and Barko 1990; Stanley, 1992), mosses and ferns (Petrini et al. 1992; Premila et al., 1996) has been recorded. They prevail in a range of tissues types within a broad range of plants, colonizing the plant systemically with bacterial/fungal colonies.Endophytes can offer several benefits to the host plant, particularly growth promotion and protection from pathogens; and that under diverse environmental conditions bacterial endophytes are able to communicate and interact with the plant more efficiently than rhizospheric bacteria (Ali et al., 2012, Coutinho et al., 2015).They are ubiquitous, colonize most of the plants, and have been isolated from almost all plants examined till date. Endophytes are to live symbiotically within the plants. While growing inside the plant the endophytes shows no visible symptoms of infection and disease (Bacon and White, 2000). The term endophyte was first carried out by De Barry in 1879. There is general consensus that microbial communities make significant contributions to plant health. Large numbers of substances are derived from the plants and therefore serve as an important source for hunting biologically active endophytes. Endophytes can also be useful for the crop production. Biological strategieshave to be developed to increase crop productivity and to reduce the usage of chemical fertilizers and also environmental stress tolerance of plant. And these can be accomplished by the judicious application of strains of plant growth promoting endophytes (PGPE)(Saraf et al.,2018). Clay &Schardl (2002) recognized three types of clavicipitaceousendophytes, ranging from symptomatic and pathogenic species (Type I) to mixedinteraction and asymptomatic endophytes (Types II and III, respectively). Endophytes are hyperdiverse at the species level, phylogenetically rich, ecologically important, evolutionarily dynamic, and represent an under explored trove of taxonomic, genetic, and functional diversity. Endophytes have been found in every plant and lichen species, including wild, crop and forage lants in all teristrial biomes samples so far. Endophytes play a major role in physiological activities of host plants influencing enhancement of stresstolerance, nematode and disease resistance (Carroll,1988; Hallmann and Siora 1996; Azevedo et al 2000; Strurz and Nowark 2000).

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Endophytes are rich in all terrestrial communities, boreal forests, an imperilled ecosystem of immense global importance and one in which their ecological associations, genetic diversity, and functional roles are largely unknown. Entophytes colonizing inside plant tissue usually get nutrition and protection from the host plant. In return they confer profoundly enhanced fitness to the host plant by producing certain functional metabolites. Endophyte containing plants grow faster than the non-containing ones (Cheplick et al., 1989). This effect is atleast in part due to production of phytohormones such as indole-3-acetic acid (IAA). Endophytes would have enhanced the hosts’ uptake of nutritional elements such as nitrogen (Ries et al., 2000) and phosphorus (Gasoni and Gurfinkel, 1997; Malinowski and Belesky, 1999). Endophytes are viewed as outstanding source of bioactive natural products as many of them occupy literally millions of unique biological niches, growing in diversified environmental conditions. Thus, it would appear that these bio typical factors can be important in plant selection since they may govern the novelty and biological activity of the products associated with endophytic microbes. The symptomless nature of endophyte occupation in plant tissues has prompted focus on symbiotic relationships between endophytes and their hosts. Both fungi and bacteria are the most common microbes existing as endophytes. Selective microbial forms like mycoplasma and archaebacteria have not yet been reportedto exists in plants as endophytes (Strobel and Daisy, 2003).Endophytes are viewed as outstanding source of bioactive natural products as many of them occupy literally millions of unique biological niches, growing in diversified environmental condition. Endophytes promote the growth of plant in various ways, that include secretion of plant growth regulators via phosphate solublizing activity (Walelin et al., 2004), production of siderophores (Costa and Loper, 1994), supplying biologically fixed nitrogen (James et al., 1994) and enhancing hyphal growth and mycorhizal colonization (Will and Sylvia, 1990).In addition, endophytes are also reported to supply essential vitamins to plants (Rodelas et al., 1993) ,confer protection against plant pathogenic microorganisms via production of antibiotics (Struz et al., 1999) or synthesis of secondary metabolites (Long et al., 2005).

Plant under examination:Euphorbia Caducifolia

Euphorbia caducifolia is a plant of Euphorbiaceae which includes 300 genera and 7,500 species. It is a latex yielding plant, grows abundantly in arid and semiarid regions of western and central India. The phylloclades of this species are tough, succulent and fleshy, covered with divaricated sharp spiny stipules. E. caducifolia (ECL) is used by the local inhabitants for treatment of bleeding

56 Vidya (2019) Vol : 14(2) Rathod et al wound, cutaneous eruption and other skin diseases. Looking to the medicinal importance of the plant, it was considered worthwhile to scientifically evaluate the for the wound healing activity. E. caducifolia is observed to be used in 31 different disease conditions. It is exclusively used in skin ailments, cough, asthma and body pain. It is effective against gastro-intestinal diseases, gynecological and obstetrical problems, respiratory diseases, in musculo-skeletal disorders, ENT disorders, immune disorders and as an antidote in snake-bite, scorpion bite. E. caducifolia has also been reported for its wound healing, (Goyal, Nagori and Sasmal, 2012) antimicrobial, antibacterial, antifungal (Goyal, Sasmal and Nagori, 2012) and anti nematologiacal activities (Maqbool, Hashmi and Ghaffar, (1987). Dry environments have plants with special adaptations which enable them to live on nutrient-poor soils, in high temperatures, and with a scarcity of water. One of the most important features is the ability to establish symbiosis with different microorganisms. Desert plants have several types of adaptations that help them conserve water. Like a leathery or waxy coating on the leaves and stems reduces evaporation. Thick stems or other plant parts provide water storage space. Small leaves or spines (modified leaves) reduce the surface area of the plant exposed to the sun. Spines and fine hairs reflect heat and reduce the air flow over the plant’s surface.

Phytochemicals in cactus

Endophytes are recognized as the potential sources of novel natural products for exploitation in medicine, agriculture and industry with more bioactive natural products isolated from the microorganisms (Bacon and White, 2000; Strobel and Daisy, 2003; Kumar and Sagar, 2007).Cactus species are important natural and economic resources in desert environments due to their adaptability and rich phytochemistry.These plants are rich in vitamin C, carotene and secondary metabolites such as polyphenols.(Stinzing and Carle,2005)these polyphenols includes mainly phenolic acids, flavonoids, tannins, saponins and alkaloids.These compounds are helpful to plants as well as human.Research have shown that plant based foods high in flavonoids can reduce mortality rates by 25% as well as significantly decrease the instances of myocardial infarction. They are also known to improve cardiac function, decrease anginas and cholesterol level. Phenols reduces the toxicity of streptozotocin by neuytalizing the free radicals produces in the pancreatic cells. Phytochemicals generally originated from the plant source are nothing but the bioactive

57 Vidya (2019) Vol : 14(2) Rathod et al compounds also known as secondary metabolites. There are two types of metabolites produced in plants viz. Primary metabolites and Secondary metabolites. Primary metabolites are important for the plants regular metabolism such as growth and development. Secondary metabolites produced by plants may have little need for them. These are synthesizing in almost all parts of the plants like bark, leaves, stem, root, flower, fruits, seeds etc. During past several years, phytochemicals have been used worldwide as the traditional herbal medicine. Because of this pharmaceuticals industry as well as researchers put a greater emphasis on the phytochemical studies. Also, these phytochemicals present in different plant parts are used up by the local people for healing of certain disorders. These are also widelyused in the field of agriculture. Secondary metabolites are economically important in the production of drugs, flavor, and fragrances, dye, and pigment, pesticides and food additives. Many of the drugs that are derived from the secondary metabolites are simple synthetic modifications or copies of the naturally obtained substances. Plants are the tremendous source for the discovery of new products with medicinal importance in drug development. Secondary metabolism, particularly in the possibility of altering the production of bioactive plant metabolites by means of tissue culture technology. Plant cell and tissue culture technologies can be established routinely under sterile conditions from explants, such as plant leaves, stems, roots and meristems for both the ways for multiplication and extraction of secondary metabolite in plant cell suspension cultures has been reported from various medicinal plants, and bioreactors are the key step for their commercial production. Based on this lime light, the present review is aimed to cover phytotherapeutic application and recent advancement for the production of some important plant pharmaceuticals. Secondary metabolites, though not essential for growth of an organism, play an adaptive role in functioning as the defense compound or the signaling molecule during ecological interactions and environmental stresses. Endophytic microorganisms produce low-molecular weight secondary metabolites that include antimicrobial compounds, phytohormones, or their precursors, vitamins like B12 (Ivanova et al. 2006) and B1 (Mercado and Bakker 2007), bioprotectants (Trotsenko and Khmelenina 2002). Several secondary metabolites are alkaloids, steroids, terpenoids, polyketones, flavonoids, quinols and phenols. These compounds also have important role in therapeutic applications such as anti-cancer, antioxidant, antimicrobial, anti-inflammatory, and immunosuppressive agents (Korkina 2007). Endophytes produce alkaloids

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as the secondary metabolites that have diverse biological potential as anti-fungal, anti-cancer, and anti-viral agents (Silva et al. 2007).

Phytochemicals helps plant to survive under harsh conditions, protects against herbivores and other insects. Alkaloids, phenols, saponins, tannins, terpenoids, carotenoids, flavonoids, steroids etc are known as phytochemicals.

Tannins

The term tannin was first introduced by Seguin to indicate various plants extracts which have the capacity to convert hides and skins into leather (Seguin A,1796). Vegetable tannins are important

59 Vidya (2019) Vol : 14(2) Rathod et al as retanning agent in the leather production and have been recognized as an important tanning agent in non-chrome tanning. Tannins are water-soluble phenol derivatives naturally synthesized and accumulated by higher plants as secondary metabolic products. From a chemical point of view, tannins are polyphenols with molecular weights between 500 and 3000 Da. In complexes with saccharides, alkaloids, and proteins its molecular weight can increase even up to 20,000 Da (Haslam Edwin, 1998) that exhibit characteristic reactions for phenols. The chemical structure of tannic acid depends on the plant species producing the compound. Currently, more than 8000 different tannins have been isolated and chemically characterized. However, there are certainly many more tannins with chemical structures that have not been precisely determined yet. All tannins have some common features, which enable classification of these types of compounds in two main groups, three types of hydrolysable tannins: gallotannines, ellagitannines, and complex tannins(sugars derivatives—mainly glucose, gallic acid, and ellagic derivatives) and condensed tannins(nonhydrolysable) called procyanidins containing condensed carbon chain typical for flavonoids (Khanbabaee and Van Ree, 2001). Condensed tannins are much more resistant to microbial degradation than hydrolysable tannins and exhibit stronger antibacterial, antiviral, and antifungal activity.

Gallic acid

Phenols

Plant phenolics" and "polyphenols" are secondary natural metabolites arising biogenetically from either the shikimate/phenylpropanoid pathway, which directly provides phenylpropanoids, or the "polyketide" acetate/malonate pathway, which can produce simple phenols, or both, thus producing

60 Vidya (2019) Vol : 14(2) Rathod et al monomeric and polymeric phenols and polyphenols, which fulfill a very broad range of physiological roles in plants. Higher plants synthesize several thousand known different phenolic compounds. The ability to synthesize phenolic compounds has been selected throughout the course of evolution in different plant lineages, thus permitting plants to cope with the constantly changing environmental challenges over evolutionary time. Plant phenolics are considered to have a key role as defense compounds when environmental stresses, such as high light, low temperatures, pathogen infection, herbivores, and nutrient deficiency, can lead to an increased production of free radicals and other oxidative species in plants. Both biotic and abiotic stresses stimulate carbon fluxes from the primary to the secondary metabolic pathways, thus inducing a shift of the available resources in favor of the synthesis of secondary products. Phenolic phytochemicals are known to exhibit several health beneficial activities such as antioxidant, anti-inflammatory, antihepatotoxic, antitumor and antimicrobial (Hertog, 1995; Rice-Evans et al., 1996; Middleton et al., 2000).

Caffeic acid

Flavonoids

Flavonoids are plant pigments that are synthesized from phenylalanine and generally display astonishing colours in the ûowering parts of plants. Flavonoids comprise a large group of polyphenolic compounds that are characterized by a benzo-y-pyrone structure, which is ubiquitous in vegetables and fruits. Besides the irrelevance in plants, ûavonoids are important for human health because of their high pharmacological activities as radical scavengers. It’s potential health benefits arising from antioxidant activities has been increased the interest of extraction of these phenolic compounds. In general, all ûavonoids are derivatives of the 2 phenyl chromone parent compound composed of three phenolic rings referred to as A, B, and C rings all of which exhibit various levels of hydroxylation and methylation. Flavonoids are the most common and widely

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distributed group of plant phenolic compounds, occurring virtually in all plant parts, particularly the photosynthesising plant cells. They are an integral part of both human and animal diets. Being plant phytochemicals, ûavonoids cannot be synthesized by humans and animals. Flavonoids found in animals are considered to originate from the plants that animals feed rather than being biosynthesized in situ. More than 5000 different plant-derived ûavonoids have been isolated from various plants. They are classiûed into at least 10 chemical groups. Flavanones, ûavones, isoûavonoids, ûavans (ûavanols), anthocyanins, and ûavonols are particularly common in the diet. Quercetin, kaempferol, and myricetin are the three most common ûavonols. Flavanones are mainly found in citrus fruits and ûavones in celery. Some ûavonoids have been reported to possess a variety of biological activities, including antiallergic, antiinûammatory, antiviral, antiproliferative, and anticarcinogenic activities, in addition of having effects on mammalian metabolism.

Flavone

Terpenoids

Terpenoidsarefeasibly the most structurally varied class of plant natural products. The name terpenoid or terpene derives from the fact that the first members of the class were isolated from turpentine. All terpenoids are derived by repetitive fusion of branched five-carbon units based on isopentane skeleton. These monomers generally are referred to as isoprene units because thermal decomposition of many terpenoid substances yields the alkene gas isoprene as a product. The smallest terpenes contain a single isoprene unit; as a group, they are named hemiterpenes (half-terpenes). The best- known hemiterpene is isoprene itself, a volatile product released from photosynthetically active tissues. Plant terpenoids are used for their aromatic qualities and play a role in traditional herbal remedies. Terpenoids contribute to the scent of eucalyptus, the flavors of cinnamon, cloves, and

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ginger, the yellow color in sunflowers, and the red color in tomatoes.[3] Well-known terpenoids include citral, menthol, camphor, salvinorin A in the plant Salvia divinorum, the cannabinoids found in cannabis, ginkgolide and bilobalide found in Ginkgo biloba, and the curcuminoids found in turmeric and mustard seed. Camphor and camphene are significant teroenoids. They are both naturallyoccurringterpenoidsprimarilyfoundun laurel trees (Cinnamonumcamphora). Camphor is a pungent terpenoid used primarily as a fragrance additive and in cooking recipes, while camphene, a monoterpene,is often used as a camphor substitute. It is sold as a color-less crystal with a strong, camphor-like scent, and is an ingredient in many foods, ointments, and topical creams. In Asia, amphor has long been used in the treatment of pain, swelling, and inflammation and served as a topical analgesic, providing a cooling effect for sore muscle tissue (Scott, R. Camphor and Camphene.)

Taxol

Alkaloids

Alkaloidsare a class of naturally occurring organic nitrogen-containing bases. Alkaloids have diverse and important physiological effects on humans and other animals. Well-known alkaloids include morphine, strychnine, quinine, ephedrine, and nicotine.Alkaloids are found primarily in plants and are especially common in certain families of flowering plants. More than 3,000 different types of alkaloids have been identified in a total of more than 4,000 plant species.Ancient people used medicinal plant extracts such as opium poppy (Papaver somniferum), Sarpagandha root (Rauwolfia serpentina) as purgatives, antitussives, sedatives, and treatments for a wide range of ailments, including snakebite, fever, and insanity.

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Coniine

Alkaloid-containing plants were mankind’s original "materia medica." Many are still in use today as prescription drugs. One of the best-known prescription alkaloids is the antitussive and analgesic codeine from the opium poppy. Plant alkaloids have also served as models for modern synthetic drugs, such as the tropane alkaloid atropine for tropicamide used to dilate the pupil during eye examinations and the indole-derived antimalarial alkaloid quinine for chloroquine.

Saponins

Saponins are naturally occurring surface-active glycosides produced by plants, lower marine animals, and some bacteria. Chemically, they generally occur as glycosides of steroids or polycyclic triterpenes. Because of their lyobipolar properties, they are able to interact with cell membranes and are also able to decrease the surface tension of an aqueous solution. This activity is the reason for the name "saponin", derived from the Latin word "sapo", which refers to the formation of a stable soap-like foam in aqueous solution. Saponins occur constitutively in a great many plant species, in both wild plants and cultivated crops. In cultivated crops, the triterpenoid saponins are generally predominant, while steroid saponins are common in wild plants used as herbs. Plant saponins have been shown to inhibit cholesterol absorption from the intestinal lumen in experimental animals and consequently to reduce the concentration of plasma cholesterol. The triterpenesaponins called ginsenosides are the major active constituents in ginseng.Saponins have been associated with numerous pharmacological actions, the important ones being permeabilizing of cell membrane, lowering of serum cholesterol levels, stimulation of luteinizing hormone release leading to abortifacient properties, immunomodulatory potential via cytokine interplay, cytostatic and cytotoxic effects on malignant tumor cells etc.

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Solanine

Identification methods of phytochemicals

Chromatography techniques

Chromatography is a technique where the molecules are separated based on their size, shape and charge. During chromatography analyte in solvent and move through solid phase that acts as a filtering material. As molecule proceeds further through molecular sieve it gets separated. Paper and thin layer chromatography are the chromatographic techniques which readily provides qualitative information and through which it become possible to obtain quantitative data.

Which includes paper chromatography, TLC, HPLC, GC etc.

Paper chromatography

In paper chromatography a sheet of paper is used for the inert phase. One of the advantages of paper chromatography is that separations are carried out simply on sheets of filter paper, which acts as both support as well as medium for separation. Another advantage is the considerable reproducibility of Rf (retention factor) values determine on paper. In paper chromatography, filter paper used as solid phase, which is inert phase. A sample is placed near the bottom of the filter paper. Then this filter paper is placed in chromatographic chamber with solvent. The solvent moves forwards by capillary action carrying soluble molecules along with it. Low porosity paper will produce a slow rate of movement of the solvent and thick papers have increased sample capacity.

Thin layer chromatography (TLC)

The first practical application of thin layer chromatography was given by Stahl. Compared to

65 Vidya (2019) Vol : 14(2) Rathod et al paper chromatography, the special advantage of TLC is the versatility, speedy and sensitive. TLC is an adsorption chromatography where samples are separated based on the interaction between a thin layer of adsorbent attached on the plate. The technique mostly employed for the separation of low molecular weight compounds. Gas chromatography (GC) Gas chromatography is a method for the separation of volatile compounds. In this method, species distribute between gas and a liquid phase. The gas phase is flowing and the liquid phase is stationary. The rate of migration for the chemical species is determined through its distribution in the gas phase. For example, a species that distributes itself 100% into gas phase will migrate at the same rate as the flowing gas, whereas, a species that distributes itself 100% into stationary phase will not migrate at all. Species distribute themselves partly in both phases will migrate at an intermediate rate. Gas chromatography involves a sample being vaporized and injected onto the head of the chromatographic column. The sample is then transported through the column by the flow of inert, gaseous mobile phase. The column itself contains a liquid stationary phase which is adsorbed onto the surface of an inert solid. High performance liquid chromatography (HPLC) HPLC is an analytical technique for the separation and determination of organic and inorganic solutes in any samples especially biological, pharmaceutical, food, environmental, industrial etc. Another name for HPLC is high – pressure liquid chromatography, separates compounds on the basis of their interactions with solid particles of tightly packed column and the solvent of the mobile phase. Modern HPLC uses a non-polar solid phase, like C18 and a polar liquid phase, generally a mixture of water and another solvent. High pressure up to 400 bars is required to elute the analyte through column before they pass through a diode array detector (DAD). A DAD measures the absorption spectra of the analytes to aid in their identification. HPLC is useful for compounds that cannot be vaporized or that decompose under high temperature, and it provides a good complement to gas chromatography for detection of compounds.

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Methods for detection Fourier-transform infrared spectroscopy (FTIR) Fourier- transform infrared spectroscopy is a valuable tool for the identification of functional groups present in the plant extract. It helps for identification and structure determination of the molecule. Samples for FTIR can be prepared in a number of ways. For liquid samples, the easiest is to place one drop of sample between two plates of sodium chloride. The drop forms a thin film between the plates. Solid samples can be milled with potassium bromide (KBr) to and then compressed into a thin pellet which can be analysed. Otherwise, solid samples can be dissolved in a solvent such as methylene chloride, and the few drop of solution is then placed onto a single High Attenuated Total Reflectance (HATR) plates and spectra was recorded in terms of percentage transmittance. The peaks at specific wave number were assigned by bonding and functional group as per the reference given in Varian FTIR instrument manual.

Nuclear Magnetic Resonance Spectroscopy (NMR)

Nuclear Magnetic Resonance Spectroscopy gives physical, chemical and biological properties of matter. One dimensional technique is routinely used but the complicated structure of the molecules could be achieved through two-dimensional NMR techniques. Solid state NMR spectroscopy is used for the determination of molecular structure of solids. Radiolabelled C NMR is used to identify the types of carbon are present in the compound. H-NMR is used to find out types of hydrogen are present in the compound and to find out how the hydrogen atoms are connected.

Mass spectrometry (MS)

Mass spectrometry is a powerful analytical technique for the identification of unknown compounds, quantification of known compounds and to elucidate the structure and chemical properties of molecules. Through MS spectrum the molecular weight of sample can be determined. This method mostly employed for the structural elucidation of organic compounds, for peptide or oligonucleotide sequencing and for monitoring the existence of previously characterizes compounds in complex mixtures with a high specificity by defining both the molecular weight and a diagnostic fragment of the molecule simultaneously.

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Cactus as Antioxidant

Cactus has been used in traditional folk medicine because of its role in treating number of diseases and condition including diabetes, hypertension, hypercholesterolemic, rheumatic pain, gastric mucosa and asthma, Cactus fruits contain bioactive compounds. It has been found that fruit juice contains a rich variety of natural antioxidants. An antioxidant is a molecule that inhibits the oxidation of other molecules. The free radicals produced during oxidative processes release highly reactive compounds generated in the body as byproducts of normal processes. Insufficient levels of antioxidants or inhibition of the antioxidant enzymes cause oxidative stress that may damage or kill DNA cells. Plant or animal foods contain a variety of nutrient/non-nutrient antioxidants, such as glutathione, vitamin C, Vitamin A and vitamin E. They are investigated for the prevention of disease such as cancer, coronary heart disease and even altitude sickness.

Conclusion

This research review’s purpose to help the reader understand different aspects of phytochemicals isolated from endophytes of cactus Euphorbiacaducifolia. And it is concluded that the desert plant is having Phenols, Tannins, Saponins, and Alkaloids as bioactive natural compounds. These compounds help plant to survive under harsh condition and also protects against herbivores. They are also having good dietary supplements which reduce hypertension, acts as anticancer, antiulcerogenic and antioxidant agents. Cactus is a desert plant which grows under harsh arid environment. Results from this study suggest that they are important producers of many novel compounds that are significant in the field of medicine, pharma, agriculture and industries.

Acknowledgement

The authors are thankful to the Department Microbiology and Biotechnology, Gujarat University for providing required facilities for conducting the research work, Teaching and Nonteaching staff of department and our parents for support.

References

Chaturvedi, H., Singh, V., & Gupta, G. (2016).Potential of bacterial endophytes as plant growth promoting factors.J Plant PatholMicrobiol, 7(376), 2.

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Cheplick, G. P., Clay, K., & Marks, S. (1989). Interactions between infection by endophytic fungi and nutrient limitation in the grasses Loliumperenne and Festucaarundinacea.New Phytologist, 111(1), 89-97.

Croteau, R., Kutchan, T. M., & Lewis, N. G. (2000).Natural products (secondary metabolites).Biochemistry and molecular biology of plants, 24, 1250-1319.

Gasoni, L., & De Gurfinkel, B. S. (1997). The endophyteCladorrhinumfoecundissimum in cotton roots: phosphorus uptake and host growth. Mycological Research, 101(7), 867-870. Strobel, G., & Daisy, B. (2003).Bioprospecting for microbial endophytes and their natural products.Microbiol. Mol. Biol. Rev., 67(4), 491-502.

Gupta, R. M., Kale, P. S., &Rathi, M. L. (2015).Isolation, characterization and identification of endophytic bacteria by 16S rRNA partial sequencing technique from roots and leaves of Prosopis cineraria plant.

Ingle, K. P., Deshmukh, A. G., Padole, D. A., Dudhare, M. S., Moharil, M. P., &Khelurkar, V. C. (2017). Phytochemicals: Extraction methods, identification and detection of bioactive compounds from plant extracts. J. Pharmacogn. Phytochem, 6, 32-36.

Joseph, B., &Priya, R. M. (2011).Bioactive Compounds from Endophytes and their Potential in.American Journal of biochemistry and Molecular biology, 1(3), 291-309. Joseph, B., &Priya, R. M. (2011).Bioactive Compounds from Endophytes and their Potential in.American Journal of biochemistry and Molecular biology, 1(3), 291-309.

Khan, P. S. S. V., Rajeswari, B., & Kumar, S. P. (2013).Potential of Euphorbia caducifolia Haines as a renewable source for biofuel.Indian Journal of Energy, 2(2), 99-107.

Musa, A. E., &Gasmelseed, G. A. (2012).Characterization of Lawsoniainermis (Henna) as vegetable tanning material.Journal of forest products & industries, 1(2), 35-40.Musa, A. E., &Gasmelseed, G. A. (2012).Characterization of Lawsoniainermis (Henna) as vegetable tanning material.Journal of forest products & industries, 1(2), 35-40.

Rodriguez, R. J., White Jr, J. F., Arnold, A. E., & Redman, A. R. A. (2009). Fungal endophytes: diversity and functional roles. New phytologist, 182(2), 314-330.

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Santoyo, G., Moreno-Hagelsieb, G., del Carmen Orozco-Mosqueda, M., & Glick, B. R. (2016). Plant growth-promoting bacterial endophytes.Microbiological research, 183, 92-99.

Thakur, M., Melzig, M. F., Fuchs, H., &Weng, A. (2011). Chemistry and pharmacology of saponins: special focus on cytotoxic properties. Botanics: Targets and Therapy, 1, 19-29.

Tubiello, F. N., Amthor, J. S., Boote, K. J., Donatelli, M., Easterling, W., Fischer, G., ...&Rosenzweig, C. (2007). Crop response to elevated CO2 and world food supply: a comment on "Food for Thought…" by Long et al., Science 312: 1918–1921, 2006. European journal of agronomy, 26(3), 215-223.

Wakelin, S. A., Warren, R. A., Harvey, P. R., & Ryder, M. H. (2004). Phosphate solubilization by Penicillium spp. closely associated with wheat roots. Biology and Fertility of Soils, 40(1), 36- 43.

Yao, L. H., Jiang, Y. M., Shi, J., Tomas-Barberan, F. A., Datta, N., Singanusong, R., & Chen, S. S. (2004).Flavonoids in food and their health benefits.Plant foods for human nutrition, 59(3), 113-122.

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Original Paper

ISSN: 2321-1520

The Challenges of Legal Profession in India: Revisiting the Saga of Mahipal Singh Rana Case

Parna Mukherjee¹*, Rhishika Srivastava² , Rahul Rajhans³

1. Asst. Prof. GLS Law College, Ahmedabad, India. 2/ 3. 5thSemester, GLS Law College, Ahmedabad, India. E-mail: [email protected]

*Corresponding Author

Received Date: 28-7-2019

Published Date:15-9-2019

Abstract

In law profession misconduct means an act done willfully with a wrong intention by the people engaged in the profession. It means any activity or behavior of an advocate in violation of professional ethics for his selfish ends. The 266th Law Commission of India Report, took cognizance of the ever- increasing instances of Professional misconduct, specifically from the mishap of Mahipal Singh Rana’s case.

The paper tries to dig deeper into the roots of professional misconduct in legal profession, looking for the potential reasons behind the same, and analyzing the gravity of the instances witnessed in this regard. It systematically attempts to analyze the steps taken by the legal fraternity to get a hold of this malpractice along with providing personal solutions to the same.

An in-depth analysis of the above-mentioned setbacks has been presented along with an logical conclusion of the subject matter.

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The paper has sought to resort to different reliable sources, both online and offline, inclusive of different online reports, surveys, statistics, studies, books and articles inter alia for the purpose of research, analysis, interpretation and execution of the subject matter.

Keywords: Law Commission of India, 266th Report of LCI, Professional Misconduct, Legal Malpractices.

Introduction

In the words of Roscoe Pound, "Historically, there are three ideas involved in a profession: organization, learning, and as spirit of public service."1 We can very well say that legal professions is culmination of all these their virtues mentioned by Roscoe Pound. Considering the background and the importance of law as an ethical profession, the makers of our Constitution, in Indian while designing the nation as a Federal structure of governance and Unitary in Spirit, entrusted separation of powers to the three organs of the Government, and further gave the Judiciary the power to interpret and enforce the rights of the people.

The dawn of modern legal profession in India, began with the passing of the Indian High Courts Act, 1861. The said act prescribed for the establishment of High Courts initially at Calcutta, Madras and Bombay, (which was subsequently extended to other high courts as well) through the letter patents during the British period.2 Thereafter, the respective High Courts were delegated with the power to make further rules for the enrollment and professional conduct of various advocates, attorneys and solicitors. The legal profession was considered to be a Nobel profession because it strived for the attainment of impartial justice in human society.

The Origin of Advocates Act, 1961 -its Legislative Intent

If we trace the genesis of the legislative development of legal profession in India post- independence period, it re-started with passing of the All India Bar Committee (1953)3 report along with recommendations. Thereafter, major development was initiated with the passing of the legislative enactment the Advocate’s Act, 1961. As we know, the said act is a codified statute governing the conduct of the Advocates and regulating the entire system of legal profession as well as legal education in India. It has many other regulatory bodies within its scope, including the Bar Council of India and the various respective State Bar Councils, which have been delegated the statutory 72 Vidya (2019) Vol : 14(2) Mukherjee et al

mandate pertaining to the admission of advocates, maintaining their rolls, governing the professional conduct, interalia of the Advocates enrolled under them. With the ever-increasing number of cases, the legal profession has now become an indispensable part of our society, which is also evident by the exponential increase in the number of enrollments of advocates throughout the country. Advocates today are vital part of our judicial system ranging from the various courts to tribunals, to law firms and legal departments in various organizations.

Professional Misconduct: A Slippery Slope

In an article it was stated that "Legal profession is a noble profession. It is not a business or a trade. A person practicing law has to practice in the spirit of honesty and not in the spirit of mischief making or money-getting. An advocate's attitude and dealings with his

client have to be scrupulously honest and fair,".4 The given statement, however may be directly in conflict with the existing realities of the legal profession in India in present times. Hence, this case of conflict and mismatch suggests that the existing legal framework governing the legal profession i. e, the Advocates Act, 1961 needs urgent amendments to eradicate the issues of professional misconduct by advocates. Also, one needs to introspects and deeply analyze the way such cases of misconduct are dealt by the local Bar councils or the even the court of law. The ever- rising demand and supply of justice however seen a contrast decline in its professional ethics and decorum. There has been increasing number of instances, where the advocates, for their personal benefit and financial as well as professional gains have turned the face of justice to a darker side. Both occasional and repetitive, has been the instances where in the need of stringent regulations for the legal professionals have been felt. In recent years, the role of advocates, particularly in the dispensation of justice through courts of law, has come under sharp criticism and is being viewed as an eyesore by the public.5 A news item published in the Indian Express titled ‘Laws for Lawyers’, spoke of the crumbling regulatory structure after having witnessed a decline in the conduct of advocates; and lawyers that was unprofessional and inconvenienced by a variety of instances.6 The conduct of advocates inside and outside the courtroom, with the judges, other fraternity members and the parties along with the radical increase of day to day strikes have reached a terrifying level and needs to be checked for in order to render true justice to individuals.

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The courtrooms nowadays often witness disturbing instances such as carrying arm, shoe hurling, abuses and giving threats to opponent advocates or judges by the appearing duly and qualified advocates themselves, may it be junior and senior as well. In addition to this, even in earlier occasions, various high courts had urged for the need for overhauling and making important amendments in the existing Advocates Act, for regulating the legal profession in matters related to professional misconduct. For example, the decision of the full judge bench of the Jharkhand High Court in the case of K. K. Jha Kamal &Anr. v. Pankaj Kumar &Anr.7, the full bench judgment of the Allahabad High Court in the case of Sadhna Upadhyay v. State of U.P8, to name a few. The gravest of all being the case of a very senior advocate hailing from the state of Uttar Pradesh named one Mahipal Singh Rana, which attracted the robust attention of the apex court, the media, the Law Commission as well as the entire judicial fraternity. The said case acted as catalyst agent to give rise to the recommendations of the Law Commission in its 266th Report. Thus, it is pertinent to analyses the various facets of this landmark case Mahipal Singh Rana v. State of U.P. to get a deeper insight in the issue of professional misconduct.

Saga of Mahipal Singh Rana’s Case

Mahipal Singh Rana vs State of UP9case came into focus in the 266th law commission as this case aroused a major issue of misconduct by a senior advocate not only affecting the judge but also the whole legal profession and judiciary. Facts of the case illustrates that Advocate Mahipal Singh Rana who was enrolled as an advocate in the UP-State Bar Council, during a hearing of case, threatened a presiding civil judge Shri Onkar Singh Yadav and literally challenged his authority as a judicial officer and the authority of the court of law. He lost his temperament when the said judge was passed on order against him and hence, threatened the Hon’ble Civil Judge stating that he will make sure that the respective judge will no longer remain as a judge in judiciary of Etah. He also openly challenged the authority of whole judiciary by disclosing the fact that already 15-20 cases have been filed against him and thus, he is not afraid of anyone! Such facts essentially construe as a case of serious professional misconduct and thus such default on the part of advocate also directly lowers the decorum of the judge, dignity of the profession and adversely affects the functioning of the judiciary. Such incident did not happen for only once but again in another case advocate Mahipal Singh Rana again showed such aggressive and disrespectful attitude towards

74 Vidya (2019) Vol : 14(2) Mukherjee et al the sitting judge. Repeatedly, Mahipal Singh hindered the court proceeding by his aggressive and hostile behaviors, challenging the authority of the pressing judges and violating the basic discipline of the judicial of the profession.

Aforestated facts and details accounts of the alleged misconduct were included by the said Judge Shri Onkar Singh Yadav in his compliant, being a witness himself he also represented before the Hon’ble High Court, urging for a strict action against such misconduct by an advocate is concerned. In the light of the above stated facts, the question arises why the local UP state bar council remained a silent spectator and allowed such an advocate to continue to terrorise, brow beat and bully the judicial officers? It is important to add here that under the Advocates Act,1961 section 6(c)10commands to every Bar Council itself having a primary authority to determine and dispose all cases of misconduct against any advocates existing on its roll. Thus, whenever any respective Bar Council fails to act or check the issues related to the misconduct of Advocates on its role, they need be made statutorily liable for such grave substantive as well as procedural lapses.

In its judgment the local High court at Allahabad held the said advocate guilty and made following observations; "We thus punish him underSection1211of the Contempt of Courts Act1971, with two months ’imprisonment and also impose fine of Rs.2000/-on him. In case non- payment offline he will undergo further a period of imprisonment of two weeks.12" Looking at this whole incident of serious misconduct, the most pertinent issue before us emerges that the term "Misconduct" is not clearly defined in the Advocates Act, 1961 and then how the cases relating to misconduct can be judiciously dealt by the respective Bar Councils or any court of law? Further, there is also an urgent need to review the existing provisions related to the quantum of the punishment, etc. Not only this, cases like Mahipal Singh Rana also puts a question mark on the regulatory role of the respective Bar Councils, the judiciary as well as the whole legal profession as a whole.

Verdict of the Apex Court

Further, the stubborn and habitually delinquent advocate Mahipal Singh filed as an appeal challenging the authority of the local High Court. In its verdict the apex court upheld the conviction and the direction given by the Allahabad High Court that the advocate shall not be permitted to appear in

75 Vidya (2019) Vol : 14(2) Mukherjee et al courts in District Etah until he purges himself of contempt. The Court also held that under Section 24A13 of the Advocates Act, the enrolment of the contemnor Advocate will stand suspended for two years. The Court also said that, as a disciplinary measure for proved misconduct, the license of the contemnor will remain suspended for further five years. The Court has however set aside the imprisonment imposed on the Advocate keeping in mind his advanced age14. Thus, this case stands as a glaring example of the existing professional misconduct in the legal profession in India in recent times.

The Apex Court in its observation very rightly held that there is an urgent need to review the existing provisions of the Advocates Act dealing with regulatory aspects for the legal profession. The Court also held that, what is permissible for this Court by virtue of statutory appellate power underSection38 of the Advocates Act is also permissible to a High Court under Article 226 of the Constitution in appropriate cases on failure of the Bar Council to take action after its attention is invited to the misconduct.15

The details analysis of the said case exposes existing several loopholes in legal frame work dealing with the issue of professional misconduct in legal profession in India. Firstly, the existing Advocates Act, 1961 definitely requires a suitable amendment to define and clarify the scope of terms "Misconduct" relating to legal profession. Secondly, there is also a need for marking a distinction between "Contempt" vis-a-vis a "Misconduct", as many cases of it may be a case overlapping both the issues. Thirdly, even though is the said case the respective High Court and Supreme Court dealt with the issue, but as per the mandate of the Advocates’ Act it must be primarily dealt by the respective Bar council itself, as already held in the case of Supreme Court Bar Association vs Union of India & Anr16.

It was laid down very clearly in the said case that, prior to the enactment of the Advocate’s Act, 1961, the former High courts were given the power to deal with the disciplinary matters of legal practitioners. Thus, previously High Courts were considered as the competent authority to punish the advocates for their professional misconduct. However, after the enactment of the Advocate’s Act, 1961 this jurisdiction of dealing misconduct of the advocates was entrusted to the respective Bar councils and further the All India Bar Council. As the Bar council being a multifunctional body

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with diverse mandate, the specific role to regulate misconduct cases has been entrusted the its Disciplinary Committees.

Thus, the Disciplinary Committees have authority to hold Disciplinary proceedings against the misconduct of advocates on any complaint received. Thus, vide the Advocates’ Act each respective Bar Councils has the statutory power to take cognizance of the disciplinary proceedings of its advocates and hence, there shouldn’t be any need for the state judiciary or the Apex court to get involve into such cases of misconduct again and again. It is just a sheer wastage of the valuable time of the apex court its worthy jurisdiction. Since, the said appeal completely lacked in merits and thus challenging such the judgement at the apex court was a complete wasteful exercise and loss of precious time of the court, however the observations of the court will be of paramount significant and will serve as judicial precedent for future. To avoid such wasteful judicial exercise, there should be clarity that what shall be the procedure and punishment and most importantly the jurisdiction of Bar councils to handle such cases of misconduct in future.

In its observation, it was expressed that there is a dire need for inserting stringent amendments and rules in the areas of Professional Conduct to the Advocates Act, was felt by the three judge bench of the Honorable Apex Court comprising of Justices Anil R. Dave, Kurian Joseph and Adarsh Kumar Goel, while hearing an appeal in a case of advocate’s misconduct. The decision went in line with the increasing number of cases of Professional misconduct by the Advocates, which got the highest gravity in the case of Mahipal Singh Rana v. State of Uttar Pradesh17. The bench requested the Government and the Law Commission to suggest appropriate measures in this regard.18

Subsequent Initiatives by Law Commission of India

The 266th Report of the Law Commission of India, titled ‘The Advocates Act, 1961 (Regulation of Legal Profession)’ suggesting important amendments to the Advocates Act, 1961 was submitted to the Government19. Considering the aforesaid judgment of the Supreme Court and the intensity of the matter along with the need for maintaining decorum of the Legal Profession as a whole, the Law Commission of India suggested some important changes inclusive of the need of new regulatory bodies and mechanism inter alia, in its 266th Report. The Commission also proposed a draft of the Advocate (Amendment) Bill 2017.20 On 22nd July2016, the Law Commission of India put an

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official notice inviting the suggestions from all the stakeholders as to how the system could be improved. Recommendations were asked from the Bar Council of India as well as various State Bar Councils as well. The Bar Council of India further constituted a committee headed by a former judge of Supreme Court Mr. Shivraj Patil. The Bar Council of India was of the view that in addition to the regulatory mechanism, other inter-related issues, i.e., constitution of the BCI and the State Bar Councils are also required to be revisited. The BCI made some suggestions in this regard.21

A detailed analysis dwelling into the major aspects of the amendments and changes suggested by the266th Report is summarized herein below. Considering the recommendations and the provisions of the existing Advocates Act, the Law Commission came to the inference that the conduct of the Advocates in one way or the other affects the functioning of the Courts and has the potential to increase the already existing pendency of cases. It also observed that the practice of going on strikes by the Advocates is rampant along the length and breadth of the country which further darkens the course of justice. The commission was astonished that despite the landmark judgment of the Supreme Court in Ex-Capt. Harish Uppal vs. Union of India 22 which stated that "… that lawyers have no right to go on strike or give a call for boycott, not even on a token strike. The protest, if any is required, can only be by giving press statements, TV interviews, carrying out of Court premises banners and/or placards, wearing black or white or any colour arm bands, peaceful protest marches outside and away from Court premises, going on dharnas or relay fasts etc.…only in the rarest of rare cases where the dignity, integrity and independence of the Bar and/or the Bench are at stake, Courts may ignore (turn a blind eye) to a protest abstention from work for not more than one day…"

Inspite of the decisions, the strikes have continued leading to the ever- increasing pendency of the cases. This goes against the decision of the Apex Court, which held that Right to speedy trial is an intrinsic part of Article 21 of the Indian Constitution. In the case of Hussainara Khatoon v. Home Secy., State of Bihar23; and in some other cases, it was held that the litigant has a right to speedy justice. The lawyers’ strike, however, result in denial of these rights to the citizens in the State. This was further affirmed by the apex court in the cases of Hussain & Anr. v. Union of India24, Ramon Services Pvt. Ltd. v. Subhash Kapoor25, Vishram Singh Raghubanshi v. State of Uttar Pradesh26,

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M.B. Sanghi v. High Court of Punjab and Haryana27, R.D. Saxena v. Balram Prasad Sharma28, Mahabir Prasad Singh v. Jacks Aviation Pvt. Ltd.29, M/s. Chetak Construction Ltd. v. Om Prakash30, Radha Mohan Lal v. Rajasthan High Court31 etc. The Constitution provides for an independent and efficient justice delivery system. Any delay in disposal of cases not only creates disillusionment amongst the litigants, but also undermines the Capability of the judicial system to impart justice in an effective manner.32 The Supreme Court has even disapproved any such conduct of the party resorting to dilatory tactics before the court seeking adjournments on one or other pretext and observed that the party acted in a manner to cause colossal insult to justice and to the concept of speedy disposal of cases33. In Re: Ajay Kumar Pandey34, the Supreme Court held: "No one can be permitted to intimidate or terrorize judges by making scandalous unwarranted and baseless imputations against them in the discharge of their judicial functions so as to secure orders which the litigant ‘wants’…The liberty of expression cannot be treated as a license to scandalize the court..." In another case the Bar Council of India v. High Court of Kerala35, the Supreme Court observed, "An advocate in no circumstances is expected to descend to the level of appearing to support his view in a vulgar brawl." In Re: S. Mulgaokar36, the Supreme Court observed that "public interest and public justice require that whenever there is an attack on the judge, it is scurrilous, offensive, intimidatory as well as malicious, the law must strike a blow on him as he challenges the supremacy of law by fouling the source and stream". Taking into account the gravity of the mentioned cases and issues, a suggestion was made by the commission for the setting up of an Advocates’ Grievance Redressal Committee headed by a Judicial Officer in every district37, which shall deal with the day to day or routine matters of the cases relating to professional misconduct. Expressing and comprehending the concerns, the Law Commission of India further suggested modifications to Section 24(A)38 of the Advocates Act, 1961 which provides for the disqualification of the enrolled advocates in certain specific cases. Section 24(A) of the Act provides that any person who is convicted of any offence is eligible to be enrolled as an advocate after the completion of two years from the date of completion of the sentence. It also went through Section 26(A)39 of the Act, which provides for the power of the Bar Councils to remove names of the Advocates from the roll, do not provide for any ground for removing the names of Advocates who are or

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were convicted of any offence under the Indian Penal Code. The issue was on the onset taken into account by the Honorable Gujarat High Court in the case of ‘C’ v. Bar Council of Gujarat40. Hence, the Commission submitted its recommendation for substituting Sections 24(A) and 26(A) with new provisions which would take into account the non-admittance of persons convicted for various crimes, as advocates and hence preserving the integrity and vitality of the Legal profession. On a rather stringent scrutiny, under the Bar Council of India Certificate and Place of Practice (Verification) Rules, 2015, the BCI found that as many as 33-45% of the then practicing advocates were bogus41 and were either practicing with fake degrees or with no degrees at all. Owing to this starling end, the Law Commission recommended a specific rule making power for Bar Council of India to make rules for verification of certificates of Advocates and for periodical verification of antecedents, conduct, place of practice of Advocates; and to make a data- based web-portal of all the advocates.42 Imparting Quality Legal Education Dwelling into the role of UGC and the BCI in imparting Legal Education throughout the Nation, The Law commission further suggested certain changes to the delivery of the Legal Education in the country. It suggested that Law Colleges should make their infrastructure revised and should provide for upgraded resources and Library facilities. The BCI should also take measures to innovate the practice of learning in order to maintain the spirit of Globalization and International competency among the Indian Law Students.

Pre-Enrollment training of Advocates

The Law Commission also highlighted the problem of newly enrolled advocates directly practicing in Higher Courts without any experience or learning by practicing in lower courts. This leads to the increase instances of proxy appearances by the senior counsels who send the freshers to take up the matter without due experience and knowledge. This in turn hampers the path of justice to be given to the citizens. While dealing with such a situation, the Supreme Court in Sanjay Kumar v. State of Bihar43, deprecated such practice by observing that an "arji", "farji" half-baked lawyer under the label of "proxy counsel", without any acquaintance with or authorization from the litigant

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use, abuse or misuse the process of the court under the false impression that he has a right to waste public time.

Further, in a nun precedented step, the Supreme Court in Re: Rameshwar Prasad Goyal44, held that conduct of Shri Goyal, AOR45, lending his signature for petty amount without appearing in court had been reprehensible and not worth pardoning, and he was censured. The court also put him on probation for a year.

Hence, the Law Commission also suggested an insertion of clause 7 in the Act, which empowers the Bar Council of India to make provisions for a mandatory training before the enrollment of a person as an Advocate.

Redefining Misconduct- The specificity of the term "Misconduct" under the statute of Advocates Act leaves room for doubt and hence, possible for either misinterpretation or vague judicial analysis. This is so because neither the Advocates Act nor the Bar Council of India Rules,46 defines professional misconduct. However, if we consider the definition by various literary sources, texts and judicial precedents, misconduct may be defined as; "A wrongful action and not mere error of judgment. A transgression of some w ell- established and definite rule of action, where no discretion is left. It is a forbidden act, carelessness, an unlawful behavior or neglect by which the right of a party has been affected e.g. allegation of disproportionate asset; misappropriation; and criminal breach of trust; not working diligently; an action which is detrimental to the prestige of the institution and acting beyond authority. It may be synonymous to an improper behavior or mismanagement. It is detrimental to public interest. Misconduct is to be construed and understood with reference to the subject matter and context wherein the term occurs taking into consideration the scope and object of the statute involved."4

Advocates have a duty to uphold the integrity of the profession and to discourage corruption so that justice may be secured by the citizenry in a legal manner.48 A lawyer must strictly adhere to the norms of profession which make him worthy as an officer of court49. Dignity of the judiciary is to be maintained, failing which the institution itself will collapse.50 Indulging in practices of corrupting

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the judiciary or offering bribe to the Judge 51; retaining money deposited with the advocate for the decree holder even after execution proceedings52 ; scandalizing the Judges53 ; constant absent from the conducting of cases; misappropriation of the amount paid54; attesting forged affidavit55 ; failure to attend trial after accepting the brief56 ; taking money from client in the name of the Judge57; gross negligence involving moral turpitude 58 ; indecent crossexamination63; breach of trust59; conducting fraud and forgery60 61 by the advocates, have been held to be serious misconduct by the Supreme Court. Considering the aforementioned decisions of the Courts, the Law Commission further provided a well -defined and comprehensive definition of Professional Misconduct in its 266thReport. Reviewing Regulatory Mechanism- The commission also recommended that for the attainment of both present and future needs of the Legal System, certain amendments, providing for strong and competent regulatory bodies must be made to the Advocates Act, 1961.Thus, the case of Mahipal Singh Rana may be remembered a blotch mark judicial history, however the case also gets credit at the same time for stirredthehornet’snestandforbringingoutseveralglaringissuerelatedtotheindisciplineexisting in legal profession in India in present times. The legislators, law makers and the regulators have to together collaborate to tackle such attitude of indiscipline and lack of professionalism. Thus, there is no iota of doubt that the existing law must expressly define the issue of "Misconduct" with reference to legal profession along with co-relating strict penal provisions to deal with it within the scope of existing legislation the Advocates Act as well as the Bar Council Rules. Further, all State bar councils must be made statutorily liable for implementation of the said rules without any scope for either substantive or procedural lapses. If urgent and necessary steps are not taken as advised by the apex court and the Law Commission of India, this legendary legal profession will continue to suffer from the plague of misconducts and indiscipline and will lose its real glory and importance in society.

References 1. Roscoe Pound, "What is a Profession - The Rise of the Legal Profession in Antiquity", 19

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Notre Dame L. Rev. 203 (1944), at p. 204. 2. Letter Patent of 1911. (1911). [ebook] England. Available at: http://www.legislation.gov.uk/ ukpga/1911/18/pdfs/ukpga_19110018_en.pdf [Accessed 20 May 2018]. 3. Barcouncilofindia.org. (n.d.). The Bar Council of India. [online] Available at: http:// www.barcouncilofindia.org/ [Accessed 20 May 2018]. 4. News, I. (2011). Advocate's professional misconduct unpardonable: SC - Times of India. [online] The Times of India, available at: https://timesofindia.indiatimes.com/india/ Advocateamp39s- professional-misconduct- unpardonable-SC/articleshow/11112845.cms [Accessed 20 May 2018]. 5.Archive.indianexpress.com. (2016). Laws for lawyers- Indian Express. [online] Available at: http://archive.indianexpress.com/news/laws-for-lawyers/432560/ [Accessed 20 May 2018]. 6.Supra Note 5. 7.AIR 2007 Jhar 67 8.2003 (2) AWC 1297 9.AIR 2016 SC 3302 10. Advocates Act, 1961; Section 6(c) Functions of State Bar Councils: - (c) to entertain and determine cases of misconduct against advocates on its roll 11.Section 12. Punishment for contempt of court (1) Save as otherwise expressly provided in this Act or in any other law, a contempt of court may be punished with simple imprisonment for a term which may extend to six months, or with fine which may extend to two thousand rupees, or with both: PROVIDED that the accused may be discharged or the punishment awarded may be remitted on apology being made to the satisfaction of the court. 12.AIR 2016 SC 3302 13.Advocates Act, 1961; Section 24A Disqualification for enrolment (1) No person shall be admitted as an advocate on a State roll-

83 Vidya (2019) Vol : 14(2) Mukherjee et al if he is convicted of an offence involving moral turpitude. If he is convicted of an offence under the provision s of the Untouchability (Offences) Act,1955. c.(Note: - Ins. by Act 70 of 1993, sec.7) If he is dismissed or removed from employment or office under the State or any charge involving moral turpitude. 14. KM, A. (2016): Law Commission Suggests Drastic Changes In Advocates Act: Live Law. [online] Live Law. Available at: http://www.livelaw.in/breaking-law-commission-suggests-drastic-changes-advocates-act/ [Accessed 20 May 2018]. 15. Supra Note 14. 16.Supreme Court Bar Association of India v. Union of India (1998) 4 SCC 409 17. AIR 2016 SC 3302 18. KM, A. (2016): Law Commission Suggests Drastic Changes In Advocates Act: Live Law. [online] Live Law. Available at: http://www.livelaw.in/breaking-law-commission-suggests- drastic- changes-advocates-act/ [Accessed 20 May 2018]. 19.266th Report on Regulation of Legal profession (The Law Commission of India) (2016), [eBook] Available at: http://lawcommissionofindia.nic.in/reports/Report266.pdf [Accessed 20 May 2018]. 20. Supra Note 17. 21.Supra Note 19. 22. AIR 2003 SC 739. 23. AIR 1979 SC 1360. 24.Criminal Appeal No. 509 of 2017 decided on 9 th March 2017 25.AIR 2001 SC 207 26. AIR 2011 SC2275. 27. AIR 1991 SC1834. 28.AIR 2000 SC2912.

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29.AIR 1999 SC 287. 30. AIR 1998 SC 1855. 31.AIR 2003 SC 1467 32. Syed Gulzar Hussain v. Dewan Syed Ale Ramul Ali Khan, (2014) 10 SCC 825 33. Gayathri v. M. Girish, (2016) 14 SCC 142. 34.AIR 1998 SC 3299 35.AIR 2004 SC 2227. 36.AIR 1978 SC 727. 37.Supra Note 4. 38.S.24(A) Advocates Act, 1961: Disqualification for enrolment. — (1) No person shall be admitted as an advocate on a State roll— (a) if he is convicted of an offence involving moral turpitude; (b) if he is convicted of an offence under the provisions of the Untouchability (Offences) Act, 1955 (22 of 1955); 4(c) if he is dismissed or removed from employment or office under the State on any charge involving moral turpitude. 39.S. 26(A) Advocates Act, 1961: Power to remove names from roll. —A State Bar Council may remove from the State roll the name of any advocate who is dead or from whom a request has been received to that effect. 40.(1982)2Guj LR 706 41.The Times of India, Delhi dated 23.01.2017, The Hindu dated 26.01.2017 and Hindustan Times dated 19.03.2017. 42.Supra Note 4. 43.(2014) 9 SCC 203. 44.AIR 2014 SC 850 45.Advocate on Record 46.Advocates Act, 1961; Bar Council of India Rules, 1961

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47. Black’s Law Dictionary, Sixth Edition; P Ramanatha Aiyar’s Law Lexicon, Reprint Edition 1987 at page 821; 48. N.G. Dastane v. Shrikant S. Shivde AIR 2001 SC 2028, Baldev Singh Gandhi v. State of Punjab AIR 2002 SC 1124; General Manager, Appellate Authority, Bank of India v. Mohd. Nizamuddin AIR 2006 SC 3290; Ravi Yashwant Bhoir v. Distt. Collector, Raigarh AIR 2012 SC 1339; and Vijay Singh v. State of U.P AIR 2012 SC 2840. 49. Shambhu Ram Yadav v. Hanuman Das Khatry, -AIR 2001 SC 2509 50. Noratanmal Chouraria v. M R Murli &Anr., AIR -2004 SC 2440. 51.In Re: Vinay Chandra Mishra, AIR 1995 SC2348 52.Supra note 33 53.Prahlad Saran Gupta v. Bar Council of India, AIR 1997 SC 1338 54.Dr. D C Saxena v. Chief Justice of India, AIR 1996 SC 2481. 55. D S Dalal v. State Bank of India, AIR 1993 SC 1608; and J S Jadhav v. Mustafa Haji Mohamed Yusuf, AIR 1993 SC 1535. 56.M Veerabhadra Rao v. Tek Chand, AIR 1985 SC 28. 57.S J Chaudhary v. State, AIR 1984 SC 618 58. Chandra Shekhar Soni v. Bar Council of Rajasthan, AIR 1983 SC 1012 59. In the matter of P an Advocate, AIR 1963 SC 1313; and V P Kumaravelu v. the Bar Council of India, AIR 1997 SC 1014 60. Shiv Narain Jafa v. The Hon’ Judges of the High Court, Allahabad, AIR 1953 SC 368. 61. Bapurao Pakhiddey v. Suman Dondey, AIR 1999 SC 916 62. LC Goyal v. Nawal Kishore, (1997) 11 SCC 258; and Devender Bhai Shanker Mehta v. Ramesh Chandra Vithal Dass Seth, AIR 1992 SC 1388; See also: Dr. Elbe Peter, MDS, LL. B., DCR, Professional misconduct of lawyers in India.

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Research Paper

ISSN: 2321-1520

Study and Characterization of Indole Carboxylate Derivative Synthesized Via Condensation

Nikita Mishra1, Tejal Gavit2, Ajay Katariya3, Dilip Vasava4*

*Department of Chemistry, Gujarat University, Ahmedabad, India E-mail:[email protected]

*Corresponding Author

Received Date: 25-7-2019

Published Date: 15-9-2019

Abstract High potency can be obtained when one biologically active molecule linked to another for resultant molecule. Here, cyclization is done by fused indole moiety to obtain highly potent, more specific and less toxic agent. In the present study, the reaction of phenyl hydrazine with a mixture of ethyl aceto acetate and glacial acetic acid by condensation reaction to synthesize desired moiety. The derivatives of indole carboxylate were obtained and characterized by FTIR and UV. The newly prepared compounds were screened for their antibacterial activity. The obtained molecule can be further applied for potential application and as intermediate for antibacterial active compounds. Keywords: Phenyl hydrazine, Ethyl acetoacetate, Hydrazine hydrate, Indole, antibacterial activity. Introduction

Heterocyclic compounds are of very much interest in daily life. Heterocyclic compounds have one or more hetero atoms in their structure. They may be cyclic or non-cyclic in nature. Heterocyclic compounds have a wide range of application. They also find applications as sanitizers, developers,

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antioxidants, as corrosion inhibitors, as copolymers, dye stuff. They are used as vehicles in the synthesis of other organic compounds. Heterocyclic compounds are also finding an increasing use as intermediate in organic synthesis. Very often this is because a relatively stable ring system can be carried through a number of synthetic steps and then cleaved at the required stage in a synthesis to reveal other functional groups. In the family of heterocyclic compounds nitrogen containing heterocycles are an important class of compounds in the medicinal chemistry and also contributed to the society from biological and industrial point which helps to understand life processes [1- 2].Indole, a privileged heterocyclic nucleus has diverse biological activities and inspired chemists to utilize this skeleton as bioactive moiety to synthesize new compounds possessing pharmacological and biological properties. The beauty of indole moiety is that it provides a privilege scaffold for the discovery of different kinds of more active, less toxic novel drugs with different modes of action. [3- 5].Amongst the wide range of methods available, the Fischer indole synthesis was employed to synthesis the indole compounds. This method involves the preliminary condensation of a carbonyl compound (usually a ketone, but occasionally an aldehyde) with an aryl hydrazine to form an aryl hydrazone, which is then rearranged to give the desired indole in a step that is often described as "indolisation". So the Fischer indole synthesis can be regarded as the elimination of ammonia from the aryl hydrazone of an aldehyde or ketone, by treatment with an acid or various metal and anhydrous metal salt catalysts, with formation of an indole nucleus [6-8].Phenyl-hydrazine is used to prepare indoles via the Fischer Indole Synthesis, which are intermediates in the synthesis of various dyes and pharmaceuticals. The indole ring system represents one of the most abundant and important heterocycles in nature. Indole and its derivatives have occupied a unique place in the chemistry of nitrogen heterocyclic compounds because of their varied biodynamic properties. Although indole motif is very small, it is an attention grabbing molecule and captured the attention of scientists and researchers over the years [9, 10]. Thus indole derivatives occupy a unique place in medicinal chemistry due to their wide range of pharmacological activities such as anti-inflammatory, antifungal, antimicrobial, antioxidant, antiviral, anti-tubercular, anticancer, anticonvulsant, antihistaminic and antagonistic, etc. [11-13].The reaction mechanism of fischer indole synthesis have been extensively studied and results has been summarized[14].In a typical fischer procedure, the aryl hydrazone is obtained by treating the requisite ketone (or aldehyde) with an equimolar quantity of the appropriate

88 Vidya (2019) Vol : 14(2) Vasava et al aryl hydrazine. The key indolisation sequence may then effected by moderate heat under thermal conditions, normally with acid catalysis [15-17]. Many different catalysts may be used, including mineral acids (sulphuric acid or hydrogen chloride), organic acids (trifluoroacetic acid and various sulphonic acids) and Lewis acids [18, 19]. A particularly convenient method involves the use of an acidic organic solvent, such as formic or acetic acid (or a homologue if a higher temperature than the boiling point of acetic acid is needed). In many instances, isolation of the intermediate aryl hydrazone is unnecessary; the reaction can be conducted as a "one-pot" procedure starting from equimolar quantities of the ketone or aldehyde and the aryl hydrazine, so that indolisation of the aryl hydrazone occurs in situ [20-23]. Preliminary studies showed that the direction of indolisation and the ratio of the product are greatly affected by the reaction conditions (i.e. the solvents and the catalysts); under weakly acidic conditions, such as with glacial acetic acid as the solvent without the addition of stronger acids, the more substituted ene-aryl hydrazines, predominated. The choice of catalysis is subjective and more a matter of task rather than based on critical experimentation [24- 26].Glacial acetic acid as a protic acid was employed as a catalyst in a solvent free condition for facile preparation of di(indolyl)methanes via one-pot condensation of indole with aryl or hetero aryl aldehydes[27-29]. AcOH as a mild and efficient catalyst for the promotion of the condensation reaction of indoles with aromatic aldehydes. So the choice of acid catalyst is very important. Bronsted acids such as HCl, H2SO4, polyphosphoric acid and p-toluenesulfonic acid have been used successfully while, lewis acids such as boron trifluoride, zinc chloride, iron chloride, and aluminium chloride are also useful catalysts [30-32]. Kalaskar et al. synthesized indole-3-acetic acids and evaluated them for their in vivo anti-inflammatory activity. The compound 1, 2- disubstituted-5-methoxyindole/ benz(g)indole-3-acetic acid showed significant activity[33-35]. The significant contribution of many derivatives of indole in the development of medicinal chemistry should be recognized.

The aim of present work is to synthesize Indole carboxylate derivatives with potential application and as intermediate for antibacterial active compounds.

2 Experimental

2.1 Material and Chemicals

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Phenyl hydrazine, Ethyl Aceto Acetate, Glacial Acetic acid, 40% bromine solution, HNO3, H2SO4 All chemicals used were of synthetic grade. The purity of compounds was ascertained by TLC on pre coated silica F254 plates (Merck, Mumbai, India) using iodine vapors and UV light as detecting agents. The melting points of the synthesized compounds were determined by open capillary method and are uncorrected. Melting points were determined in open capillaries in electrical apparatus and are uncorrected. IR spectra were recorded on a FT-IR spectrometer using KBR pellet.

Synthesis of Indole Derivatives

STEP-1: Synthesis of Ethyl-2-methyl-1H-indole-3-carboxylate[36] [C1]

O O O Glacial Acetic Acid + O NH Reflux temp. N NH2 O H

1-Phenyl Hydr azine ethyl 3 - oxobutanoate ethyl 2- methyl-1H- indole- 3-carboxylate

In a flat bottom flask (250 ml), a sealed stirrer unit and reflux condenser, a mixture of ethylacetoacetate (2.4 ml; 0.018 mole) and glacial acetic acid (1.0 ml,0.018mole)was placed in the flat bottom flask and heated under reflux with stirring. Add phenyl hydrazine (1.8 ml; 0.018mole) slowly during first 1hr. Continue the stirring for further 1hr. Reaction progress is checked by TLC method. Pour the reaction mixture into a 50ml beaker with cooled water and stir vigorously while it solidifies. Then add sufficient water and filter. Dry the crude product, the crude product thus obtained was recrystallized from methanol.

STEP 2: Synthesis of Ethyl5-bromo-2-methyl-1H-indole-3-carboxylate [B1]

2

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In 250 ml conical flask take Ethyl-2-methyl-1-H-Indole-3-Carboxylate (2 gm, 0.0098mole) dissolve in suitable solvent. i.e. Methanol. Put this solution in ice bath to maintain temperature 0- 10°C. Take 40% Br solution (2.5 ml) and transfer it into separating funnel. Then add this Br solution dropwise to above solution at temperature maintaining 0-10°C. After addition put the mixture at R.T for 20 min. then dump this into ice water, stir vigorously so the product is obtained.

Step 3: Synthesis of Ethyl 6-nitro-2-methyl-1H-indole-3-carboxylate [N1]

O O O O Co nc. HNO3 + Con c. H 2SO 4 O2N

o 0-5 C N N H H

ethyl 2-methyl-1H-indole-3-carboxyl ate ethyl 2-methyl-5-nitro-1H-indole-3-carboxylate

Take HNO3 (1 ml, 0.019 mole) in one test tune and add, in portions with shaking, another test tube containing concentrated H2SO4 (2 ml, 0.039 mole). Keep the mixture cool during the addition by immersing the test tube in cold water. Then add this mixture dropwise in RBF containing dissolved Ethyl-2-methyl-1-H-Indole-3-Carboxylate (2 gm, 0.0098 mole) in methanol. Try to maintain temperature by immersing RBF in cold water. When all nitrating reagent mixture added, fix the reflux condenser to RBF; Heat in water bath to maintained temperature 60°-70°C for 30 minutes. Then pour this into cold water to get the product. 2.3 Properties and Characterization Techniques 2.3.1 SOLUBILITY [37] Solubility in the different solvents depends upon the solvent type and on the functional groups if present in the solution. Neutral organic compounds tend to be hydrophobic; that is, they are less soluble in water than in organic solvents. Organic compounds tend to dissolve in organic solvents. The solubility of compounds can be determined by placing approximately 0.1gm of finely grind polymer in test tube with 5ml of particular solvent for each. The mixtures were stored at 25°C for some time with continues shaking. While shaking, formation of streaks indicated dissolution. Samples that swelled without dissolving at 25°C were heated to 50°C in order to affect the process of dissolution.

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2.3.2 UV-VIS SPECTROSCOPY[38]

Ultraviolet-visible spectroscopy involves the absorption of ultraviolet/visible light by a molecule causing the promotion of an electron from a ground electronic state to an excited electronic state. A beam of light from a visible and/or UV light source (colored red) is separated into its component wavelengths by a prism or diffraction grating. Each monochromatic (single wavelength) beam in turn is split into two equal intensity beams by a half-mirrored device.

Ultraviolet/Visible light: wavelengths between 190 and 800 nm

Ultraviolet range between 190-380 nm&

Visible region falls between 380-750 nm

UV-Vis Spectroscopy is the measurement of the attenuation of a beam of light after it passes through a sample or after reflection from a sample surface. Absorption measurements can be at a single wavelength or over an extended spectral range.

UV-Vis spectroscopy is routinely used in the chemistry for the quantitative determination of different analytes, such as transition metal ions, highly conjugated organic compounds, and biological macromolecules. Solvent polarity and pH can affect the absorption spectrum of an organic compound.

2.3.3 FT-IR SPECTRA [39]

It is a very popular method infrared spectroscopy is being utilize for characterizing samples. Vibration of atoms of the molecules is main concept based on this technique. Through a sample IR radiation are passed in which IR spectrum is obtained with an energy. IR spectrum explains characteristic of the entire molecule, in which certain groups of atoms give rise to bands at or near the same frequency regardless of the structure of the rest of the molecule.

2.3.4 ANTIMICROBIALACTIVITY [40]

In recent years, there has been a growing interest in researching and developing new antimicrobial agents from various sources to combat microbial resistance. Therefore, a greater attention has been paid to antimicrobial activity screening and evaluating methods. Several bioassays such as

92 Vidya (2019) Vol : 14(2) Vasava et al disk-diffusion, well diffusion and broth or agar dilution are well known and commonly used. Antimicrobial susceptibility testing can be used for drug discovery, epidemiology and prediction of therapeutic outcome.

Dilution methods are the most appropriate ones for the determination of MIC values, since they offer the possibility to estimate the concentration of the tested antimicrobial agent in the agar (agar dilution) or broth medium (macro dilution or micro dilution). Either broth or agar dilution method may be used to quantitatively measure the in vitro antimicrobial activity against bacteria and fungi. MIC value recorded is defined as the lowest concentration of the assayed antimicrobial agent that inhibits the visible growth of the microorganism tested, and it is usually expressed in µg/mL or mg/ L.

3 Results and Discussion

3.1 Physical Properties

Name of Compound Mol. formula M.W. M.P. Color (gm/mole)

Ethyl-2-methyl-1H-indole-3- C12H13NO2 203.24 135°C Cream carboxylate yellow

Ethyl 5-bromo-2-methyl-1H- C12H12BrNO2 282.13 127°C Yellowish indole-3-carboxylate Orange

Ethyl 6-nitro-2-methyl-1H- C12H12N2O4 248.23 194°C Yellowish indole-3-carboxylate Brown

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3.2 Solubility Chart Sr. No. Name of Compound Solubility

1 Methanol ++

2 DMF ++

3 THF ++

4 Hexane +-

5 Toluene +-

6 CHCl3 ++ 7 MDC ++

8 Ethyl Aceto acetate ++

9 Acetone ++

10 DMSO ++

11 Acetonitrile ++

12 CCl4 --

13 Water --

3.3 IR Analysis

Figure 1: IR analysis of C1

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Figure 3: IR analysis of B1

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Interpretation of IR Compound C-H C-N N-H C=C C-X C=O N-O C-Mono -1 -1 -1 -1 -1 -1 -1 (cm ) (cm ) (cm ) (cm ) (X=Br) (cm ) (cm ) Substituted(CH3)(cm ) (cm-1) C-1 3124 1163 3109 1533 - 1591 - 908 B-1 3124 1163 3109 1533 661 1591 - 908 N-1 3124 1163 3109 1533 - 1591 1548 908

3.4 UV Analysis

Figure 4: UV analysis of C1

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Figure 5: UV analysis of N1

Figure 6: UV analysis of B1

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Interpretation of UV Molecule Wavelength of Maximum Absorption (nm) C1 250nm N1 217nm B1 248nm A jump from a pi bonding orbital to a pi anti-bonding orbital ought to have a fixed energy and therefore absorb a fixed wavelength.

All of the molecules give similar UV-visible absorption spectra - the only difference being that the absorptions move to longer and longer wavelengths as the amount of delocalization in the molecule increases.

3.5 Antibacterial Activity Analysis

The newly prepared compounds were screened for their antibacterial activity two gram- positive bacteria S. aureus and two gram-negative bacteria E.coli using Nutrient agar medium.

Antibacterial activity was carried out by Disc-diffusion method [zone of inhibition] and serial dilution method [MIC].Antibacterial activities were studied by subjecting the compounds to pharmacological screening by standard procedures.

All the compounds synthesized in the present investigation were tested for their antibacterial activity. The antibacterial activities were tested on nutrient medium against Staphylococcus aureus and Escherichia coli.

• Investigation: Antimicrobial activity of B2 sample:

Method: Agar Diffusion

Sr. No. Test organism Sample Name- bromo-B2

100 µg 50 µg 25 µg

1 E. coli 100 mm NO ZONE NO ZONE ATCC 25922 OBSERVED OBSERVED 2 S. Aureus 20 mm 15 mm 14 mm ATCC 25923

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• Investigation: Antimicrobial activity of C2 sample:

Sr. No. Test organism Sample Name- C-2

100 µg 50 µg 25 µg

1 E. coli 10 mm NO ZONE NO ZONE ATCC 25922 OBSERVED OBSERVED 2 S. Aureus 10 mm NO ZONE NO ZONE ATCC 25923 OBSERVED OBSERVED

Conclusion

The indole derivatives are abundantly available in wide variety of natural sources and new indole derivatives are being synthesize on regular basis. The synthesized Ethyl 5- bromo 2-methyl-1-H- indole-3-carboxylate showed enhanced biological activities. This synthesized indole carboxylate derivatives are analyzed by IR and UV analysis. They showed a broad spectrum of antibacterial activity against G(+ve) and G(-ve) bacteria. It may be worthwhile to explore the possibility in this area by fusing other heterocyclic moieties and increase the potency of synthesized compounds.

Acknowledgement

I would like to express my deepest gratitude to the Head, Department of Chemistry, Gujarat University for providing the facility of UV-analysis. I am also thankful to my guide for their constant support and encouragement. I also thanks to The Director, ultrasonic laboratory, Ahmedabad for IR analysis. I greatly acknowledge the Director, Biocare research pvt. Ltd. Paldi for Antibacterial Activities.

References

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Arora P, Arora V, Lamba HS and Wadhwa D, Int J Pharm Res Sci. 3(9); 2947-2955, (2012). Ashcroft, C.P.; Hellier, P.; Pettman, A.; Watkinson, S. Second-Generation Process Research Towards Eletriptan: A Fischer Indole Approach. Organic Process Research and Development, 15, 98-103, (2011).

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Brodfuehrer, P.R.; Chen, B.; Sattelberg, T. R.; Smith, P. R.; Reddy, J. P., Stark, D.R.; Quinlan, S. L.; Reid, J. G.;. J. Org. Chem., 62, 9192-9202, (1997). Douglass F. Tabera, and Pavan K. Tirunahari, Tetrahedron, 67(38): 7195–7210, (2011). Goldberg R, kishore, N.; Lennen, R.; J. of Physical and Chemical Reference Data, 31 (2): 231-370; (2002). Hickman, A. B.; Klein, D. C.; Dyda, F., Molecular Cell, 3, 23-32, (1999). I.-K. Park, S.-E.Suh, B.-Y.Lim, C.-G.Cho, Org. Lett.; 11, 5454-5456, (2009). J. Acta Chemica Scandinavica, 43,651-659, (1989). Kalaskar, G. P.; Girisha, M.; Purohit, M. G.; Thippeswamy, B. S.; Patil, B. M. Indian J Heterocycl Chem ; 16, 325, (2007). Kelly, A. H.; McLeod, D. H.; Parrick, J., Canadian Journal of Chemistry, 43, 296-301, (1965). M.A. Pfaller, D.J. Sheehan, J.H. Rex; Clin. Microbiol. Rev., 17, 268-280, (2004). Mardia El-Sayeda,b , Kazem Mahmouda and Andreas Hilgeroth, Current Chemistry Letters 3 , 7–14, (2014). Martyn Inman and Christopher J. Moody, J. of Royal Society of Chemistry, Chem. Sci., 2013, 4, 29–41, (2013). Miller, B., Pearson Education, Inc: Upper Saddle River, NJ,pp 372-373, (2004). Mills, K.; Al Khawaja, I. K.; Al-Saleh, F. S.; Joule, J. A., Journal of the Chemical Society, Perkin Transactions 1: Organic and Bio-Organic Chemistry, 636-4, 1972-1999, (1981). Nagendra Kumar Kaushik, Neha Kaushik, Pankaj Attri ,Naresh Kumar et al,. Molecules, 18, 6620-6662, (2013). O. Miyata, Y. Kimura, K. Muroya, H. Hiramatsu, T. Naito, Tetrahedron Lett., 40, 3601-3604, (1999). P. BharathRathna Kumar, S. Subramaniyan, K. Yamini and R. Suthakaran, 4 (2), 400-400, (2011). Panda SS, Chowdary PVR. Indian j pharma Sci; 70,208-215; (2008) Pausacker, K. H., Journal of the Chemical Society, 621-4, (1950). R. D. Brown, B. A. Coller, Aust. J. Chem., 12,152 (1959). R. J. Sundberg, Academic Press, New York (1970). Robinson, B., Chem. Rev. 373-401, (1963).

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Roussel, P. A. The Fischer Indole Synthesis. J. Chem. Educ., 30, 122-125, (1953). S. Gore, S. Baskaran, B. König, Org. Lett., 14, 4568-4571, (2012). S. Wagaw, B. H. Yang, S. L. Buchwald, J. Am. Chem. Soc., 120, 6621-6622, (1998). Sami Sajjadifar, HooshangVahedi, Abdolhossien Massoudi and Omid Louie, J. Molecules, 15, 2491-2498; (2010). Schomerus, C.; Korf, H. W., Annals of the New York Academy of Sciences, 1057, 372-383, (2005). Shriner, R.L.; Hermann, C.K.F.; Morrill, T.C.; Curtin, D.Y. and Fuson, R.C.; John Wiley & Sons, ISBN 0-471-59748-1, (1997). Sosale Chandrasekhar &Somnath Mukherjee, An International Journal for Rapid Communication of Synthetic Organic Chemistry, 45(8), 1018-1022, (2015). Standard Practice for Describing and Measuring Performance of Ultraviolet, Visible, and Near- Infrared Spectrophotometers, ASTM Standard E 275-83, (1994). Stoermer, D.; Heathcock, C. H., Journal of Organic Chemistry, 58, 564-8, (1993). Streitweiser, Andrew Jr; Healthcock clayton H; macmillian,Isbn 0-02—418010-6, (1976) Sundberg, R.J. Indoles. Academic Press: San Diego; pp 1-2, 54-55, (1996). Sureyya Olgena, Nurten Altanlarb, Ersin Karataylý , and Mithat Bozdayýc; Z. Naturforsch. 63c, 189-195, (2008). Urban, M. W., Encyclopedia of Analytical Chemistry, Vol. 3, Meyers, R. A. (Ed.), Wiley, Chichester, UK, pp. 1756–1773, (2002). Van Order, R. B.; Lindwall, H. G., Indole. Chem. Rev, 30, 69-96, (1942). W. J. Houlin, Indoles; Wiley-Interscience: New York, Part I (1972). Wagaw, S.; Yang, B. H.; Buchwald, S. L., Journal of the American Chemical Society 121, 10251- 10263 (1999).

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Review Article

ISSN: 2321-1520

A Review on Sugar and Citric Acid as Chemical Preservatives for Cut Flower

Christian Stacey1*, Ekta Patel2, Ruby Patel3 and Himanshu Pandya4

1, 2, 4Department of Botany, University School of Sciences, Gujarat University, Ahmedabad 3President Science College, Gujarat University, Ahmedabad E-mail: [email protected]

*Corresponding author

Received Date: 27-6-2019

Published Date: 15-9-2019

Abstract Maintaining good quality of cut flowers and extending the vase life, are considered important and practical for having acceptable products for the markets. For this reason, a considerable number of studies have been undertaken for this purpose. Vase life of cut flowers is affected by many factors. The prevalent method for maintaining the vase life of cut flowers is the use of moderately low temperatures. An alternative is the use of chemical preservatives. Such a solution can affect vase life, ethylene production, and regulation of sugar accumulation in floral organs. Treatment of cut flowers with sucrose and citric acid is found to be beneficial in delaying senescence processes. Keywords:Cut flowers, Chemical preservatives, Sugar, Citric acid. Introduction Cut flowers are precious products of horticulture. The post-harvest longevity of cut flowers is of critical importance in determining the value of the crop. Maintaining good quality of cut flowers and extending the vase life, are considered important and practical for having acceptable products for the markets. For this reason, a considerable number of studies have been undertaken for this

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purpose. (Macnishet al., 2008 and Solgiet al., 2009, Zencirkiran, 2010).Many investigations on the longevity and quality of cut flowers have been conducted by adding various preservatives to the vase water (Reid, 2012), resulting in cut flowers senescence being delayed considerably. In cut flowers, the processes of flower bud opening and colour development require substrates and energy for their satisfactory development (Uthaichayet al., 2007). Factors affecting vas life According to Kelegama (2001) Factors such as the mode of transportation, condition during transport, storage at the distributors and time lag from shipment to ultimate sale are very important factors in the maintain the quality and extending the shelf life.Vase life of cut flowers is mainly affected by two main factors, namely ethylene which accelerates the senescence of many flowers and microorganisms which cause vascular blockage and thus reduce the vase life of cut flowers (Van Doorn, 1994, Zencirkiran, 2010).As described by Wilberg (1973), flower wilting can be considered as one of the main postharvest disorders which may lead to stem break that occurs 10 cm below capitulum. As well as this, blockage of xylem vessels due to bacterial or microorganisms accumulation is another contributing factor leading to quality loss (JaliliMarandi, et al., 2011). This blockage can be culminated in water uptake deficiency and water loss (Hassan, 2005). Role of preservatives A floral preservative usually is a complex mixture of sucrose (sugar), acidifier, an inhibitor of microorganisms and also an ethylene action or synthesis inhibitor like STS and SA (Tehranifaret al., 2013). The prevalent method for maintaining the vase life of cut flowers is the use of moderately low temperatures. An alternative is the use of chemical preservatives. Such a solution can affect vase life, ethylene production, and regulation of sugar accumulation in floral organs (Ichimura and Hisamatsu, 1999). Sucrose: Ichimura and Hiraya (1999) reported that the pulse treatment with 100 g L-1 sucrose in combination with 200 mg L-1 HQS for 16 h had a significant effect on extending the vase life of cut sweet pea flowers.Aarts (1957) also suggested that exogenous sucrose in some way maintains the structure and semi-permeability of the plasma membrane. Furthermore, treatment of cut flowers with sucrose is found to be beneficial in delaying senescence processes (Chung et al., 1997).Many studies have reported on the application of sucrose in extending the vase life of cut flowers. For example, Paulin and Jamain (1982) and Kaltaler and Steponkus (1976) demonstrated the vase life of cut carnations and roses, respectively, increased following sucrose treatment. Similar results

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were found in cut flowers of Chrysanthemum(Patel et al., 2016a) and Tithonia (Patel et al., 2016b). In our study, the increase in vase life by the pulse treatment with sucrose was significant in comparison to water controls.Meir et al. (1995) reported that mini-gladiolus cut spikes, together with sucrose plus STS pulsing, offered potential advantages of extending their vase life and maintaining flower quality. Moreover, Han (1998) also reported that the postharvest quality of cut Heucherasanguinea was significantly improved and its vase life significantly increased by pulsing the inflorescence with STS for 4 h followed by placing the stems in a sucrose solution containing cntdotL-1 8-hydroxyquinoline citrate. Citric acid:It is a widespread organic acid in the plant kingdom and makes a weak acid in water. Citric acid is used to adjust water pH and to control the growth of microorganisms. Citric acid is commercially advised for a number of cut flowers like chrysanthemum (Dole and Wilkins, 1998). Also, it reduces the risk of vascular blockage in cut flowers through its anti- embolism trait. Citric acidtreatments extended vase life in association with inhibition of ethylene production (Srivastava, 2000). Citric acid is also known as an acidifier which inhibits the growth of microorganisms and is commercially advised for a number of cut flowers including chrysanthemum (Dole and Wilkins, 1998). It can alleviate water uptake and extend vase life due to its anti-embolism trait (Bhattacharjee et al., 993). Water constitutes a large proportion of horticultural products weight. In addition to water, carbohydrates are the other major constituent of these products. These products commonly take water and other materials from the mother plant, but when cut off, they rapidly move into senescence and death which take place of water loss and weight reduction. This reduction is much higher in stress conditions. Conclusion The vase life of cut flowers and foliage is often shortened by vascular occlusions that constrict vase solution supply. Cut flower and foliage longevity can be greatly affected by the chemical composition of the vase solution. External sugars can be provided to cut flowers by dissolving a known amount of sugar, along with a biocide, into the vase solution. Most flowers benefit from a continuous supply of 2% sugar in the vase solution. Citric acid is used to lower the pH. It has been shown that low pH water (pH=3.5) travels faster in the water-conducting system (xylem), thereby preventing or reducing wilting that frequently occurs in field-grown flowers. References Aarts, J.F.T. (1957): On the capability of cut flowers. Meded.Landbouwhogesch.Wageningen

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57: 1-62. Bhattacharjee, S. K., Singh, V., and Saxena, N. K. (1993): Studies on Vegetative Growth, Flowering, Flower Quality and Vase Life of Roses. Singapore J. of Primary Industries. 21 (2): 67-71. Chung, B.C., S.Y. Lee, S.A. Oh, T.H. Rhew, H.G. Nam, and C.H. Lee. (1997): The promoter activity of sen 1, a senescence-associated gene of Arabidopsis, is repressed by sugars. J. Plant Physiol. 151: 339-345. Dole, J. M., F. Harold and P. Wilkins. (1998): Floriculture Principles and Species. Prentice Hall Inc, Upper Saddle River, New Jersey. 613P. Han, S.S. (1998): Postharvest handling of cut HeucherasanguineaEngelm. flowers: Effects of sucrose and silver thiosulfate. HortScience 33: 731-733. Hassan, F., (2005): Postharvest studies on some important flower crops. Ph.D. Thesis, Faculty of Horticultural Sciences, Corvinus University of Budapest, Hungary. Ichimura, K. and T. Hiraya (1999): Effect of siver thiosulfate complex (STS) in combination with sucrose on the vase life of cut sweet pea flowers. J. Japan. Soc. Hort. Sci. 68: 23-27. Ichimura, K. and T. Hisamatsu (1999): Effects of continuous treatments with sucrose on the vase life, soluble carbohydrate concentrations, and ethylene production of cut snapdragon flowers. J. Japan. Soc. Hort. Sci. 68: 61-66. JaliliMarandi, R., Hassani, A., Abdollahi, A., Hanafi, S., (2011) : Improvement of the vase life of cut gladiolus flowers by essential oils, salicylic acid and silver thiosulfate. Journal of Medicinal Plants Research, 5 (20), 5039-5043. Kaltaler, R.E.L. and P.L. Steponkus, (1974): Uptake and metabolism of sucrose in cut roses. J. Amer. Soc Hort. Sci., 99: 490-493. Kelegama, S. (2001): Agriculture and the new trade agenda in the WTO 2000 negotiations: Economic analysis of interests and policy option for Sri Lanka. (www.unctad.org/trade_env/ test1/meetings/standards/shrilanka.doc) Macnish, A. J., Leonard, R. T and Nell, T. A. (2008): Treatment with Chlorine Dioxide Extends the Vase Life of elected Cut Flowers. Postharvest Biol. Technolo.50: 197-207. Meir, S., S. Philosoph-Hadas, R. Michaeli, and H. Davidson (1995): Improvement of the keeping quality of mini-gladiolus spikes during prolonged storage by sucrose pulsing and modified atmosphere packaging. Acta Hort. 405: 335- 342.

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Patel, R., Parmar, M., Bhatt, M. and Mathur, M., (2016a): Effect of chemical preservatives on shelf life of Cosmos sulphreus cav. Cut flower, International Journal of Advanced Research, 4(11), 1467-1471. Patel, R., Thakkar A, and Mankad A., (2016b): Effect of chemical preservative on water relation and vase-life of Tithonia rotundifolia Blake Cut flower, Annals of biological Research, 7 (1):27- 30. Paulin, A. and C. Jamain. (1982): Development of flowers and changes in various sugars during opening of cut carnations. J. Amer. Soc. Hort. Sci. 107: 258-261. Reid, M.S.,(2012): Handling of cut flowers for air transport. http://ucce.ucdavis.edu/file/ datastore/234 1373.pdf. 05/09/12. Solgi, M., Kafi, M., Taghavi T. S and Naderi, R. (2009): Essential Oils and Silver Nanoparticles (SNP) as Novel Agents to Extend Vase-Life of Gerbera (Gerbera jamesonii cv. ‘Dune’) Flowers. Postharvest Biol. Technol.53: 155-158. Srivastava, M. K. and Dwivedi, U. N. 2000.Delayed Ripening of Banana Fruit by Salicylic Acid.Plant Science, 158: 87-96. Tehranifar A, Vahdati N, Selahvarz Y, Bayat H, (2013):Treatment with salicylic acid extends the vase life of important commercial cut flowers, Advanced Crop Science, 3 (6): 405–413. Uthaichay, N., S. Ketsa and W.G. Van Doorn (2007): 1-MCP pretreatment prevents bud and flower abscission in Dendrobium orchids. Postharv.Bio.and Techno., 43: 374-380. Van Doorn W.G., Zagory, D., Witte, Y.D. and Harkema H. (1994): Effect of Vase-water Bacteria on the Senescence of Cut Carnation Flowers. Postharvest Biol. Technol., 1:161-168. Wilberg, B., (1973): PhysiologischeUntersuchungenzumknickert-problem alsVoraussetzungfür die selektionhaltbar Gerbera-Schnittblumen. Z. Pflanzenzucht. 69, 107-114. Zencirkiran, M. (2010): Effect of 1-MCP (1- Methyl Cyclopropene) and STS (Silver thiosulphate) on the Vase Life of Cut Freesia Flowers. Sci. Res. Essay.5 (17): 2409-2412

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Research Paper

ISSN: 2321-1520

Cultivation and Study of Growth of Oyster Mushroom on Different Agricultural Waste

Shukla Rachana M. 1, Shrimali Priyanka2 and Thakkar Aarti3*

President Science College, Gujarat University, Ahmedabad, Gujarat, India E-mail:[email protected]

*Corresponding Author

Received Date:2-7-2019

Published Date: 15-9-2019

Abstract The study was conducted to compare the effects of different agro-wastes on the growth and yield of oyster mushroom Pleurotusostreotus. Cultivation of Pleurotusostreotus on different substrate such as sugarcane, paddy straw and sorghum. Sugarcane produced highest yield, biological efficiency and number of fruiting bodies, recommended as a best substrate for oyster mushroom cultivation. The highest number of primodia and fruiting bodies and the amount of fresh weight was obtained with sugarcane bagasse in all flushes whereas, the lowest with paddy straw and sorghum. Keywords: cultivation, Oyster mushroom, Substrates, Sugarcane baggase, Yield Introduction Mushrooms are one kind fungus which may be edible or inedible. Edible fungi belong to the genus Pleurotus under the class ofBasidiomycetes. Oyster mushroom (Pleurotusostreatus) is one of the edible mushrooms having excellent flavour and fine taste. Pleurotus species are popular and widely cultivated throughout the world mostly in Asia and Europe because of their simple and low

107 Vidya (2019) Vol : 14(2) Thakkar et al cost production technology and higher biological efficiency [1]. Cultivation of oyster mushroom have increased tremendously throughout the world now-a-days because of their abilities to grow at a wide range of temperature and they can be harvested all over the year [2]. Pleurotus have the ability to excrete hydrolyzing and oxidizing enzymes [3]. Pleurotusostreatus demands few environmental controls and their fruiting bodies are not often attacked by diseases and pests and they can be cultivated in a simple and economic way [4]. It requires a shorter growth time in comparison to other edible mushrooms [5]. Mushrooms require carbon, nitrogen and many different inorganic compounds as their nutritional sources. The main nutrients are less nitrogen and more carbon congaing substrate such as most organic matters containing cellulose, hemicelluloses and lignin can be used (i. e. rice and wheat straw, cottonseed hulls, corncob, sugarcane baggase, sawdust, waste paper, leaves and so on) which supports the growth, development and fruiting of mushroom [6]. The growth of diverse type of mushrooms require different type of substrates (organic or inorganic) and availability of varied type of materials that may dictate which type is used [7]. In general edible mushrooms are low in fat and calories but rich in vitamins B, D, K, A and C [8]. They contain more protein than any other food of plant origin and are also a good source of mineral nutrients [9]. Malnutrition is a major problem in developing third world countries. Mushrooms with their flavour, texture, taste, nutritional value and high productivity per unit area have been identified as an excellent food source to alleviate malnutrition in developing countries [10]. Oyster mushroom cultivation can play an important role in managing organic wastes whose disposal has become a major problem worldwide [11]. These wastes can be recycled into an environment and may be less endangered by pollution [12]. Strengthening mushroom production sector could be essential in order to enable the rural economy to keep its vibrancy and development, increasing and diversifying business which beings employment opportunities in the rural areas as well as providing income opportunities to small family farms. Furthermore, the use of these residues in bioprocesses technology also may be one of the solutions to bioconversion of inedible biomass residues into nutritious protein rich food in the form of edible mushrooms [13]. Apart from food value, its medicinal value for diabetics and cancer therapy has been emphasized [14]. Many of mushrooms pose a range of metabolites of intense interest to pharmaceutical sector e.g. antitumour metabolites, antigenotoxic metabolites, antioxidant metabolites, anti-inflammatory metabolites, anti-

108 Vidya (2019) Vol : 14(2) Thakkar et al hypertensive metabolites, antipalletel aggregating metabolites, anti-hyperglycaemic metabolites, antimicrobial metabolites, antiviral activities and food industries [15]. Classification of Oyster Mushroom:

Scientific classification:

Kingdom – Fungi

Phylum – Basidiomycota

Class - Agaricomycetes

Order - Agaricales

Family - Pleurotaceae

Genus – Pleurotus

Species – ostreatus

Materials and Method

Materials

1. Spawn (Source:- M. G. Science Institute, Ahmedabad, Gujarat)

2. Sugarcane baggase, Paddy straw as raw materials.

3. Water bath

4. Hot Air Oven

5. Plastic Bags

6. Poly propylene bottles

7.

Chemicals

1. 1N NaOH

2. Chloroform : Methanol (2:1)

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3. 1% Gypsum

4. Potato Dextrose Agar

5. 1% Calcium super phosphate

Methods

Microorganism and spawn preparation:

The methodology that was used in these experiments was standardized following the methods of Siddiqui [16].Pleurotusostreatus, obtained from the Mushroom Spawn was grown on the potato dextrose agar (PDA 200 g/l diced potatoes; 20 g/l glucose; 15 g/l agar) medium at 250C for regular subculture. Oyster mushroom spawn was prepared in 850-ml polypropylene plastic bottles filled with cotton seed hull 87%, wheat bran 10%, sucrose 1%, plaster stone 1% and calcium superphosphate 1% (w/w, in terms of dry weight), then sterilized in autoclave at 1210C for 80 min. After cooling down to room temperature, 1% gypsum was added. Then sterilized cotton seed hulls of every bottle were inoculated with 5 cm2 mycelial agar discs. The spawn was incubated in the laboratory at 26 ± 20C and 70% relative humidity for two weeks.

Image: Spawn Preparation

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Substrate preparation:

Oyster mushroom was grown on various substrates viz. Paddy straw, sugarcane baggase and sorghum. All the substrates were chopped into 3-4 cm lengths with cutter and then completely sun dried.

Soaking: All three substrates were then soaked in water with 1-5 ppm formaldehyde overnight before the final step performed for substrate preparation. Physico-chemical methods were used for the sterilization purpose. Heat treatment was given to each and every substrate for avoiding contamination problems. Such treatment also helps in giving higher and almost constant yields from particular substrate used. Widely used method is pasteurization. Pasteurization: Water was boiled in a wide mouth container (steel/aluminium). The wet substrates were filled in gunny bags before they get dry. The filled bags were dipped in hot water (approximately 700 C) for about 10-15 min. After pasteurization, excess hot water was drained off from container so that it can be reused for other sets and thus temperature was also maintained. Spawning: Pasteurized substrates were cooled down to room temperature. At this stage, moisture content maintained at constant degree in all substrates at about 70%. Polythene bags were used for cultivation. Bags of 25cm×15cm in size were used. About 2.5 % grain spawn were inoculated on the surface of substrate and incubated in a controlled dark ttemperature around 25-350 C. The bags were closed after spawning. Humidity maintained at 70-85% by spraying water twice a day on walls and floor. Cropping and harvesting: After 10-15 days of incubation, bags were fully impregnated with white mycelium. Polythene covers were removed for further analysis. Mushroom was kept in a temperature range of 20-330 C (Mesophillic range). Relative humidity was maintained by spraying water twice a day on the wall and floor. A light spray of water was given on blocks as soon as small premordia appeared. Once

111 Vidya (2019) Vol : 14(2) Thakkar et al premordia appeared, watering to blocks was stopped to allow them to grow. After harvesting, the mushroom weight was recorded to calculate the biological efficiency.

Weight of fresh mushroom Biological efficiency = ------× 100% Dry weight of substrate

Total weight of harvested fresh mushroom per bag Mushroom weight = ------Total number of mushrooms harvested per bag

After harvesting, the mushrooms were packed in perforated polythene bags to keep them fresh till evaluation. It loses freshness after about 6 hours, which can be enhanced by keeping them in refrigerator. Oyster mushroom can be shed dried for 2/3 days and dried product are marketed in labelled polythene bags.

Results

Number of primordial:

The highest numbers of primordia (20 primordia) were found in sugarcane baggase and the lowest found in sorghum waste. The similar trends were observed in the second and third flush.

Figure 1. Primordial in sugarcane waste Figure 2. Primordial in Paddy straw Number of fruiting bodies: The highest number of fruiting bodies of oyster mushroom was found in sugarcane baggase (20) and lowest numbers found in sorghum waste in 1st flush. Numbers decreased at second and third flush.

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Figure 3. Fruiting bodies Fresh weight of fruiting bodies: The highest fresh weight of fruiting bodies was obtained in sugarcane baggase, whereas the lowest found in sorghum. These results were found statistically different in different substrates.

Figure 5. Harvested oyster mushroom

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Table 1. Comparison between three different substrates

Discussion

In the first flush, the number of primordia was found with significant variation in all substrates. The number of fruiting bodies was found to vary significantly among all substrates. The similar kind of results was observed in the 2nd and 3rd flush.

For each of the treatments, two flushes of mushroom were harvested. The yield was significantly highest obtained in the first flush of all treatments used. The study revealed that the yield for sugarcane baggase was highest yield among all treatments, but it was not significantly highest. Paddy straw was also almost at par. Growth on sorghum waste was also at par with sugarcane baggase.

Sugarcane baggase gave maximum number of primordia and fruiting bodies and fresh weight followed sorghum. The variations were probably occurred due to the difference of bulk density and constituents of the substrates used in study [17]. found that the yield of mushroom was correlated positively with cellulose content and cellulose: lignin ratio, but negatively with lignin and ortho- dihydroxyphenolics content remarkably [18].Stated that cellulose rich organic substances have been reported to be of good substrates for the cultivation of this kind of edible mushrooms.

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Substrates with high lignin and phenolic content decreased the activity of cellulose enzyme, but less lignin would enhance enzyme activity and thus ensure higher yield of mushrooms grown by such method [19]. Mshandeteet.al. (2008) reported that sugarcane baggase contains both cellulose and sucrose content which is easily degraded by oyster mushrooms. It is also rich in nitrogen content, which is favorable for mushrooms [13].

Conclusion

Mushroom cultivation is one of the efficient ways by which residues can be recycled. P. ostreatusgrown on different substrates are nutritious with high protein content, fiber content and low fat content. It may also offer economic incentives for agribusiness to examine these residues as valuable resources and develop new enterprises to use them to produce nutritious mushroom products. Therefore, the mushroom cultivation may become one of the most profitable agribusiness which could produce food products from different agricultural wastes or other substrates and help to dispose them in an environment friendly manner.One can conclude from the results of above experiments that sugarcane baggase have higher nutritional value in comparison of other substrates used. It is one of the cheapest sources of nutrition for mushroom cultivation.

Acknowledgements

The authors highly acknowledge the Dr. MrugeshShukla, M.G. Science Institute, Navrangpura, Ahmedabad, Gujarat, India. They gave training and facilitated premises as well as ground for mushroom cultivation.

Reference

A. Eswaran, R. Ramabadran, Studies on some physiological, cultural and post-harvest aspects of oyster mushroom, Pleurotuseous, Tropical Agricultural Research, 12, 2000, 360 – 374.

A. Eswaran, R. Ramabadran, Studies on some physiological, cultural and post-harvest aspects of oyster mushroom, Pleurotusostreatus, Tropical Agricultural Research Journal, 12, 2000, 360- 374.

A.M. Mshandete, J. Cuff, Cultivation of three types of indigenous wild edible mushrooms: CoprinusCinereus, Pleurotusflabellatus and Volvariellavolvocea on composted sisal decortications

115 Vidya (2019) Vol : 14(2) Thakkar et al residue in Tanzania, Afr. J. Biotech., 7(24), 2008, 4551-4562.

H. Sivrikaya L. Bacak A. Saacbasi, I. Toroguli, H. Erogulu, Trace elements in PleurotusSajor- caju cultivated on chemithermomechenical pulp for bio-bleeching, Food Chem., 79, 2002, 173- 176.

L. Pathmashini, V. Arulnandh, R. S. W. Wijerathan, Efficancy of different spawn types on sawdust media (Tropical Agricultural research and Extension, 2008) 11.

N. Bahl, Hand book on mushrooms (Oxford & IBH Publishing co Pvt Ltd., 1998), pp.15-40.

N. Caglarirmak, The nutrients of exotic mushrooms (Lentinulaedodes and Pleurotus species) and an estimated approach to the volatile compounds, Food Chem., 105, 2007, 1188–1194.

N. Das, M. Mukherjee, Cultivation of Pleurotusostreatus on weed plants. Bio Resource Technology, 98, 2007, 2723 – 2726.

Quimio, T. H. 1987. Introducing Pleurotusflabellatusfor your dinner table.The mushroom Journal, 96: 282-283. S. M. R. Amin, C. S. Nirod, M. Moonmoon, J. Khandaker, M. Rahman, Officer’s Training Manual (National Mushroom Development and Extension Centre, Savar, Dhaka, Bangladesh, 2007) 7-17.

S. T. Chang, Mushroom cultivation using the "ZERI" principle: potential for application in Brazil, MicologiaAplicadaInternatonal, 19(2), 2007, 33-34.

S. T. Chang, P. G. Miles, Edible Mushroom and their cultivation (CRC press, Inc. Boca Raton, Florida U.S.A. 1988) 27: 83-88.

Siddiqui, A. B. Mushroom Production Technology. IHAND project, GOB/UNDP/ FAO, Rangdhanu Printers, 2002, pp. 1-14.

Sivaprakasam, K. 1980. Studies on Oyster mushroom (Pleurotussajor-caju) Fr. Ph. D. Thesis, Tamil Nadu Agril.Univ., Coimbatore, India.

Sivaprakasam, K. 1986. Constituents of substrates in relation to sporophore yield of

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Pleurotussajor-caju Fr. Madras J. Agril., 73(11): 601-605.

T. Kausar, Cultivation of mushrooms using crop residues as substrate, Ph.D. Thesis. Department of Botany, University of Punjab, Lahore, Pakistan, 1988.

U. Kues, Y. Liu, Fruiting body production in basidiomycetes, Appl. MicrobiolBiotec, 54, 2000, 141-152.

V. P. Mane, S. S. Patil, A. A. Syed, M. M. V. Baig, Bioconversion of low quality lignocellulosic agricultural waste into edible protein by Pleurotussajor-caju (Fr.) Singer. Journal of Zhejiang University of Science, 8(10), 2007, 745-751.

Z. A. Shah, M. Ashraf and Ch. Ishtiq, Comparative study on cultivation and yield performance of oyster mushroom (Pleurotusostreatus) on different substrates (Wheat straw, Leaves, saw dust). Pak. J. Nutri., 3, 2004, 158-160.

117 Vidya (2019) Vol : 14(2)

Review Article

ISSN: 2321-1520

Virtual Laboratories in STEM Courses: A Critical Review

Bhumika Shah*, Jyoti Pareek

Department of Computer Science,Gujarat University, Ahmedabad. Email : [email protected]

*Coresponding Author

Received Date: 14-8-2019

Published Date:15-9-2019

Abstract

Computer Science courses demand extensive laboratory work as well as strong subject knowledge. Information technology resources have given wings to innovations and advancements by providing an artificial educational environment, which can be used along with the traditional environment. Such virtual environments provide many advantages like students are not required to install costly software or need not have specific set up for the desired course, just an internet connection, and a browser can serve the purpose. There are very few systems equipped with virtual laboratory experiments, along with tutorials related to the subject. The computer science courses particularly lack this type of learning management systems (LMS). Most of the Virtual laboratories reviewed are either just simulator-based or are some remote environments accessed through the cloud. Majority of the Virtual laboratories exist in Basic Sciences and other Engineering domains, whereas Computer science courses have very limited virtual laboratories.

Introduction

E-learning is a concept which evolved as online learning and utilized different forms of electronic

118 Vidya (2019) Vol : 14(2) Shah et al technologies, unlike a traditional classroom concept. Initially, e-learning resources were used as assisted teaching in classrooms for displaying videos, animations, etc. for better understanding of the concept. E-learning platforms are becoming critical platforms not only for educational institutions but also for corporates and individuals for lifelong learning

Web technologies are reshaping the education systems, and the innovations in technology have the ability to take education systems to new dimensions. The Learning component, Teaching-Learning methods, Flipped learning, Blended learning are all the significant researched areas in the education systems.Each Year (Educause) surveys the Higher education community to determine the areas of improvement in the Teaching and Learning Process.

The picture in Figure 1 represents key issues of Teaching and learning for the year 2019.

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These types of surveys help the Academicians to address the teaching-learning process and bring the technological advancements with a clear focus on innovative pedagogical approaches. The evidence of flipped learning’s effectiveness is nonetheless plentiful in the area of educational research since 2014. 2017 alone had 131 peer-reviewed articles on flipped learning. (Robert Talbert). However, the term flipped learning is rapidly getting attention from the research communities in varied forms and approaches. Some researchers believe that just using technological aids gives the teaching a label of flipped learning, whereas there are researchers who have insisted that classroom-based learning is inverted to deepen the understanding among students which helps to make the process of understanding more effective. Modern Re-usability frameworks assume that a learning object is a moveable entity (Brusilovsky & Nijhawan, 2002), Data which is stored in a repository and can be reused by copying on the destination client. But in advanced re-usable objects, activities are delivered by a Web Server(Brusilovsky & Nijhawan, 2002) which need to reside permanently on a web server and can be accessed by multiple clients. Here, researchers suggest that the learning activities should be adaptive based on the environment and student needs. It promotes courseware reuse and adaptive educational systems. They propose a knowledge tree where learning goal or objective is adaptive with the learning need of the student. (Olin, Bourne, Mayadas, & Consortium, 2005) believe that Learning effectiveness, student satisfaction, Faculty satisfaction, access, and cost-effectiveness are the five metrics that drive investigations into online education. Computer Science is one discipline that uses the growing and demanding e-Learning environment to deliver the computation and technology and knowledge to the learners. Computer science needs practical sessions as the main part of its education process(Fechner, Keller, & Schiffmann, 2005a). The engineering bodies for accreditation (ABET) also strongly recommend for lab sessions/ practical sessions in their evaluation criteria. Laboratory work is an integral part of computer science courses. The rise of technological advancements has led to innovations and improvement in the online educational system, including

120 Vidya (2019) Vol : 14(2) Shah et al online learning courses. The innovative approaches help the courses to be designed as per the changing requirements of the next generation learning. To achieve effective learning, laboratory work should be integrated with e-Learning environments. For courses like Computer Science laboratory work is an integral part of learning, thus providing a virtual laboratory environment for computer science courses can prove beneficial.

As our focus is on computer science domain, the limited duration for Lab sessions and fixed lab time slots in most of the colleges make the lab work limited or restricted to a particular time slot. The need of today’s era is to let the student's practice/implement lab work in their pace and time. To provide such a facility, a separate infrastructure in the form of remote laboratories is required.

Virtual laboratories offer flexibility, allowing students to visit laboratories at odd times and enabling them to revise concepts as and when needed. Additionally, Virtual laboratories are beneficial in situations where the students are deprived of the laboratory infrastructure or have insufficient access to the laboratory resources.

Table 1: Traditional Laboratories Vs Virtual Laboratories Traditional Laboratory Virtual Laboratory Fixed Time and Place Any time and Place, based on the user’s requirement Expensive Less Expensive compared to Traditional Laboratories Learning and Assessment are done as per Complete Learning Management traditional methods of teaching System(LMS) at one place All students addressed in one group at one Personalized Learning time Less student engagement Higher student engagement Teacher/Learner-centric Pedagogy Learning Centric Pedagogy

The Virtual Laboratories highly emphasize on Student Engagement. The digital platform brings with them the limitation of being diverted easily from the focus area. Hence, if the developers want a student to spend more time in a virtual laboratory, and practice and learn more, then the system developed

121 Vidya (2019) Vol : 14(2) Shah et al should be more engaging. The system needs to focus on learning-centric pedagogy to achieve the same. The experiments should be designed in such a way that students enjoy learning, which could be achieved by properly defining the Learning Objectives. The Learning objectives, in turn, should be aligned with the cognitive levels which help in catering the needs of all types of students and engaging all types of students. The appropriate cognitive level selection helps in defining the task correctly, and hence, the assessment gets adequately represented.

The joint ACM-IEEE Curriculum Task Force (1989) recommends thatcomputer science courses should be supported by extensive laboratory work.

In computer science courses, laboratories are an integral part of the learning experience. The instructors must consider all the objectives of the subject while designing tasks for laboratory- related work.The Learning objectives provide a clear idea about the outcome expected from the course getting designed.As, In today’s world, students are turning towards e-Learning environment, it becomes essential that e-Learning environments are able to integrate virtual laboratories to provide complete learning.

RELATED WORK

There are many e-Learning repositories and courses available on e-Learning contents. However, the e-Learning contents mainly focus on delivering content to a mass audience and allow sharing of pools. These type of learning environments do not provide student interaction, and their primary focus is good quality e-content/tutorials. However, laboratory work is equally important, along with good quality course content. With the increasing technological advancements, even the laboratory work is required on the e-Learning platform, which results in the laboratories being available remotely. Hence, there is a need for Virtual Laboratories. However, good quality Virtual Laboratories are rare. The computer science virtual laboratories are even lesser. Virtual Laboratories in education are referred to as the simulation environment that adds an environment and human touch to the virtual interfaces (Virtual Laboratory, 2019). The primary purpose of a Virtual Laboratory is to provide remote access to laboratories in the area of science and engineering. The virtual environment gives an advantage of sharing costly equipment’s which otherwise might be limited to a few users due to constraints on time and geographical distances. Such virtual

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environments provide the benefit of addressing a large number of students at once, and the subject expert develops the content at one location, which can be accessed by students in different geographical areas. The virtual laboratory research is still in its infancy stage, and there is an acute dearth of good quality virtual laboratories.

A whole heap of research already exists on e-Learning systems and different online learning materials or learning courses. However, all those e-Learning contents involve delivering the content only; very few of them include live student interaction or student engagement. In the past,researchers have suggested that the learning activities should be adaptive based on the environmentand student needs. It promotes courseware reuse and adaptive educational system

Computer Science courses are not entirely instruction-based, and most of the subjects demand extensive practical work along with the conceptual clarity of the said subject. The traditional laboratories are the solutions for the implementation of the concepts learned.However, with the growing Information technology needs and innovative learning environments, the demand is to have laboratory access at their own pace, time, and place. Plenty of research isconductedto provide the students with the traditional laboratory environment on a virtual platform while also making the entire experience more interactive.

(Bourne, Harris, & Mayadas, 2013)suggests that the e-learning tools should be comparable with the traditional classrooms. They believe that learning effectiveness, student satisfaction, faculty satisfaction, access, and cost-effectiveness are the five metrics that drive investigations into online education.

Many researchers have argued that the students have started losing interest in traditional eLearning curriculum. The area of Learning requires significant innovations, wherein the student’s interest can be maintained and adhere to the learning capacity of the students concerning different learning levels of the students. However,STEM (Science, Technology, Engineering, and Maths) courses still demand more efforts in learning skills. The engineering or computer science course is not complete without the laboratory work.Hence, there is a demand for the virtual/remote laboratory which can satisfy student’s need for practical implementation in the digital content. However, to make laboratory learning effective and maintain student’s interest, the pedagogical perspective has

123 Vidya (2019) Vol : 14(2) Shah et al a vital role to play. The pedagogical effectiveness helps design the course with all the learning objectives in mind, which results in a learning-centric virtual laboratory getting developed.

(Fechner, Keller, & Schiffmann, 2005b) discuss that Computer Science is a discipline that uses the growing and demanding e-Learning environment to deliver the computation, technology, and knowledge to the learners. Computer science needs practical sessions as the central part of its educational process.

In other engineering disciplines, researchers have proposed the following virtual laboratories.

(Smith, 2004)introduced an online simulator for Microelectronic devices. They designed and implemented an MIT device simulation WebLab (WeblabSim). It was an online simulator wherein the students could explore the behavior of microelectronic devices. WeblabSim makes device simulator readily available anywhere, anytime.

In August 2005, (Kantzavelou, 2005)proposed a virtual lab model for introductory computer science courses. The model consists of seven modules, and each one corresponds to a specific topic of the course. Every module provides several different services to a particular topic of the course. Every module offers various services to assimilate theory and practical exercises. The proposed model was partially implemented and gave a better understanding of theoretical concepts

(Kuleshov, 2008) Rightly points out that the interactive simulations allow students to explore a topic by comparing and contrasting different scenarios. Users may get more in-depth exposure to the subject matter either by modifying parts of existing simulator or by building a new simulation from scratch.

In 2015, (Al-Khanjari & Al-Roshdi, 2015)proposed the usage of Service Oriented Architecture (SOAs) to overcome the limitation of Learning Management Systems towards the practical implementations in computer science education. The developer uses SOA to develop the required resources as services and then integrates them in the LMS.

In February 2016, (Mendes, Li, Bailey, Delong, & Del Alamo, 2016)proposed a web services approach to support interactive LabVIEW based Remote experiments under MIT’s iLabShared Architecture(ISA). Experiment lab server Architecture (ELSA) was developed as an extension to

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ISA, fulfills the requirements of an ISA process agent and uses web services to connect the client with the lab Server.

In November 2016, (Velasquez, Ramos, & Amaya, 2016) reviewed different technologies which could be used for the development of different types of virtual/remote laboratories in the education system. They conclude that the Remote Laboratories (LRS) lack interactivity and there are many technologies available which could be integrated to develop aninteractive LR. They also explored various disadvantages of techniques which require compulsory installation of add-ons as a prerequisite. Such considerations should be eliminated to create an interactive and open platform LR/VR.

Additionally, there are other educational areas which are also explored concerning Virtual laboratories like Languages, Gamings, etc. One of the language lab named as "The Virtual Language Lab," which helps the students to learn a foreign language in a virtual environment. (Mads Bo- Kristensen, 2008)insist that even before the virtual lab was implemented pedagogically and practically, new significant options of Mobile language lab had emerged. And even before the latest lab displayed its pedagogical potential, yet another lab scenario was ready in the form of pervasive computing and learning.

Understanding the need of user interaction for effective learning, MHRD initiated Virtual Lab project. Virtual labs project is collection of many virtual laboratories consisting of different experiments in the field of Science and engineering. The project is anconsortium of many Participating institutes and IIT-Delhi is the coordinating institute. The Virtual laboratories are available in the areas like Electronics and Communication, Biotechnology and Biomedical , Computer Science, Civil engineering, Physical sciences , Electrical, Mechanical and chemical engineering. The new labs developments include many innovative areas and technologies. One of the major advantage of virtual laboratories is they are not just simulator based , but they are the entire Learning Management System at one place.

Virtual lab project is dedicated to address the basic problem of lack of good lab facilities in many engineering colleges. (Bose, 2013)

Virtual labs is more than just providing laboratory access to students , It enhances the students

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thinking capacity and motivates him to learn more. As(Bose, 2013) rightly mentions that good teachers are always a scarce resource. Hence, such Virtual lab project is an Open Educational Resource (OER) that addresses the issue of lack of good lab facilities, as well as trained teachers by making remote experimentation possible.

The Virtual lab project is intended to benefit varied communities like Students and Faculty members , High-school students whose inquisitiveness would be triggered, Researchers, who can collaborate inter-institute or inter-discipline and Different engineering colleges. (Bose, 2013)

Conclusion

Authors could review that various remote or virtual laboratories are mostly based in engineering disciplines. Most of the models proposed were simulator-based and provided access to the program on a remote computer or a server.Authors believe that a virtual laboratory proves more efficient if it caters to all the needs of a complete Learning Management System and provides step-by-step guidance in the process of learning.

The virtual laboratories available are just simulator based and not able to provide guided learning. There is need to create complete LMS, considering all the Teaching Learning aspects such as pedagogy of teaching and learning, student engagement and cognitive levels. All the cognitive levels from Recall to Create should be considered to facilitate higher order of learning to students. The virtual laboratories should be such that students should seek to spend more time and should feel that they can learn more by using the virtual laboratory. The online labs under the Virtual labs project not only arouse curiosity in students but also permit them to learn at their own pace.

Many virtual laboratories have been developed and are being developed, but there is lot of scope of improvement in virtual lab design and development. Also sometimes E-Learning platforms are not as effective as traditional classroom based learning, because of one major factor.In classroom based approach a teacher can teach with different methods and different pace to each student according to their grasping capabilities. Hence, Development of smart virtual lab which can give personalized learning experience to each student as per their pace of learning and needs is an open issue for researchers to explore.

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References Al-Khanjari, Z., & Al-Roshdi, Y. (2015). Developing virtual lab to support the computer science education in moodle. Proceedings of 2015 12th International Conference on Remote Engineering and Virtual Instrumentation, REV 2015, (February), 186–191. https://doi.org/ 10.1109/REV.2015.7087290 Bourne, J., Harris, D., & Mayadas, F. (2013). Online Engineering Education: Learning Anywhere, Anytime. Journal of Engineering Education, 94(1), 131–146. https://doi.org/10.1002/j.2168- 9830.2005.tb00834.x Brusilovsky, P., & Nijhawan, H. (2002). A Framework for Adaptive E-Learning Based on Distributed Re-usable Learning Activities. 154–161. Retrieved from http://www2.sis.pitt.edu/ ~peterb/papers.html Fechner, B., Keller, J., & Schiffmann, W. (2005a). Evaluation of a Virtual Computer Architecture Lab. Kantzavelou, I. (2005). A virtual lab model for an introductory computer. Facta Universitatis (Nis), Ser.: Elec. Energ., 18(2), 263–274. Mendes, L. A., Li, L., Bailey, P. H., Delong, K. R., & Del Alamo, J. A. (2016). Experiment lab server architecture: A web services approach to supporting interactive LabVIEW-based remote experiments under MIT’s iLab shared architecture. Proceedings of 2016 13th International Conference on Remote Engineering and Virtual Instrumentation, REV 2016, (February), 293–305. https://doi.org/10.1109/REV.2016.7444486 Olin, D., Bourne, J., Mayadas, F., & Consortium, S. (2005). Online engineering education/ : Learning anywhere , anytime . Online Engineering Education/ : Learning Anywhere , Anytime. Journal of Engineering Education, 9(January). Smith, A. C. (2004). MIT Device Simulation WebLab/ : An Online Simulator for Microelectronic Devices. Velasquez, Y., Ramos, O., & Amaya, D. (2016). Technology used for the implementation of remote laboratories. International Journal of Applied Engineering Research, 11(11), 7456– 7461.

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Bose, R. (2013). Virtual Labs Project: A Paradigm Shift in Internet Based Remote Experimentation. IEEE Access, (pp. 1-8) Educause. (n.d.). Retrieved from https://www.educause.edu/ Educause. (n.d.). Issues in Teaching and Learning. Retrieved from https://www.educause.edu/ eli/initiatives/key-issues-in-teaching-and-learning Mads Bo-Kristensen, B. M. (2008). Transformations of the Language Laboratory. Handbook of Digital Information Technologies , IGI GLobal. O'Sullivan, M. (Nov 2004). The Reconceptualisation of Learner-Centred Approaches: A Namibian Case Study. International Journal of Educational Development, v24 n6 , p585-602. Robert Talbert, P. (n.d.). Myths and Facts about Flipped learning. Retrieved from https:// er.educause.edu/articles/2017/9/myths-and-facts-about-flipped-learning Shah, B. (2017). Open Source Databases. Retrieved from E-pgpathshala: https:// epgp.inflibnet.ac.in/view_f.php?category=1490 Vavrus, F. (2009). The cultural politics of constructivist pedagogies: Teacher education reform in the United Republic of Tanzania. International Journal of Educational Development, 303- 311. Virtual Laboratory. (2019). Retrieved from EduTech Wiki: http://edutechwiki.unige.ch/en/ Virtual_laboratory

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Review Article

ISSN: 2321-1520

Review on Heavy Metal Toxicity on Plants.

Hemangini Gohil *, Jahnavi Pandya & Archana Mankad

Department of Botany, Bioinformatics & Climate Change Impacts Management School of Science, Gujarat University, Ahmedabad 380 009 E-mail :[email protected]

*Corresponding Author

Received Date: 19-6-2019

Published Date: 15-9-2019

Abstract

Heavy metals are the intrinsic component of the environment with essential and non essential both types. Although many metal elements are essential for the growth of plants in low concentrations, their excessive amounts in soil above threshold values can result in toxicity. Soils polluted with heavy metals have become common due to increase in geologic and anthropogenic activities. It is the unplanned disposal of municipal waste, mining, use of extensive pesticides, insecticides, fungicides, and other agrochemicals uses were significant causes of environment pollution and causes of most concern.

Keywords:Heavy metals, Toxicity, Anthropogenic, environmental pollutants, Threshold.

Introduction

Heavy metals are deûned as the elements having density greater than 5 g cm"3 (Adriano 2001). Some heavy metals namely, cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), nickel (Ni) and zinc (Zn) are considered to be essential for plants, whereas chromium (Cr),

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and antimony (Sb) are found essential for animals (Misra and Mani 1991; Markert 1993). These metal elements can directly inûuence growth, senescence and energy generating processes due to their high reactivity. Their concentration in soil beyond permissible limits is toxic to plants either causing oxidative stress through free radicals and/or disrupting the functions of enzymes by replacing essential metals and nutrients (Henry 2000; Prasad 2008). Although changes in cell metabolism permit plant to cope with, yet the reduction in plant growth is the primary symptom of metal toxicity.

Among the heavy metals, chromium and cadmium are of special concern due to their potential toxicity to both animals and plants even at low concentrations (Sharma et al. 1995; Das et al. 1997; Shukla et al. 2007). The chromium toxicity in plants varies from the inhibition of enzymatic activity to mutagenesis (Barceloet al. 1993). The visible symptoms include leaf chlorosis, stunting, and yield reduction (Das et al. 1997; Boonyapookana et al. 2002). Cadmium (Cd) is particularly dangerous pollutant due to its high toxicity and great solubility in water (Pinto et al. 2004). Cadmium also inhibits the uptake of elements such as K, Ca, Mg, Fe because it uses the same transmembrane carriers (Rivettaet al. 1997). Its accumulation in plants may also pose a serious health hazard to human beings through food chain; however, it poses an additional risk to children by direct ingestion of Cd-contaminated soil (Nordberg 2003).

Effect on Growth and Development

Heavy metals either retard the growth of the whole plant or plant parts (Shaûq and Iqbal 2005; Shankeret al. 2005). The plant parts which have the direct contact with the contaminated soils normally the roots exhibit rapid and sensitive changes in their growth pattern (Baker and Walker 1989). Heavy metals mainly affect plant growth through the generation of free radicals and reactive oxygen species (ROS), which pose constant oxidative damage by degenerating important cellular components (Pandey et al. 2005, Qureshi et al. 2005). The typical symptoms of Cd toxicity of rice plants are wilted leaves, growth inhibition, progressive chlorosis in certain leaves and leaf sheaths, and browned root systems, especially the root tips (Das et al. 1997; Chugh and Sawhney 1999). In addition, in maize (Zea mays) Cd also reduces plant growth (Talanovaet al. 2001; Liu et al. 2003/2004).

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Conclusion

Several heavy metal elements are essential for biological and physiological functions of plants, including biosynthesis of proteins, nucleic acids, growth substances, synthesis of chlorophyll and secondary metabolites, stress tolerance, structural and functional integrity of various membranes and other cellular compounds. However, beyond permissible limits, these metal elements become toxicdependinguponthenatureandspeciesofmetalandplants.Metaltoxicitymayinhibitelectrontransport,

reduce CO2 ûxation, and cause chloroplast disorganization. Heavy metal stress can induce a series of events in plants leading to decrease in number and size of leaves, enhancement of leaf rolling and leaf abscission changes in stomatal size and resistance.

References Adriano DC (2001), Trace elements in terrestrial environments. Biochemistry, Alburry, Australia, pp 1–16. Baker AJM, Walker PL (1989), Physiological responses of plants to heavy metals and the quantitiûcatioin of tolerance and toxicity. Chem Spec Biovail 1:7–17 Barceló J, Poschenrieder C, Vazquez MD, Gunse B, Vernet JP (1993), Beneûcial and toxic effects ofchromiuminplants:Solutionculture,potandûeldstudies.StudiesinEnvironmentalScience No. 55, Paper Presented at the 5th International Conference on Environmental Contamination. Morges, Switzerland. Boonyapookana B, Upatham ES, Kruatrachue M, Pokethitiyook P, Singhakaew S (2002), Phytoaccumulation and phytotoxicity of cadmium and chromium in duckweed ,Wolfûaglobosa. Int J Phytoremed 4:87–100 Chugh LK, Sawhney SK (1999), Photosynthetic activities of Pisumsativum seedlings grown in the presence of cadmium.Plant Physiology Biochem. 37(4):297–303 Das P, Samantaray S, Rout GR (1997) Studies on cadmium toxicity in plants: a review. Environ. Pollut. 98:29–36. Henry JR (2000),InAn overview of phytoremediation of lead and mercury. NNEMS Report Washington, pp 3–9.

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Liu DH, Jiang WS, Gao XZ (2003/2004), Effects of cadmium on root growth, cell division and nucleoli in root tip cells of garlic. Bio. Plant 47(1):79–83.

Misra SG, Mani D (1991) Soil pollution.

Nordberg G (2003), Cadmium and human health: a perspective based on recent studies in China. J Trace Elem Exp Med 16:307–319.

Pinto AP, Mota AM, de Varennes A, Pinto FC (2004), Inûuence of organic matter on the uptake of cadmium, zinc, copper and iron by sorghum plants. Sci. Tot Environ. 326:239–247.

Prasad MNV (2008), Trace Elements as Contaminants and Nutrients: Consequences in Ecosystems and Human Health. Wiley, New York.

Qureshi MI, Israr M, Abdin MZ Iqbal M (2005), Responses of Artemisia annua L. to lead and salt Pandey V, Dixit V, Shyam R (2005) Antioxidative responses in relation to growth of mustard (Brassica juncea cv. Pusa Jai Kisan) plants exposed to hexavalent chromium. Chemosphere 61:40–47 induced oxidative stress. Environ. Exp. Bot. 53:185–193

Rivetta A, Negrini N, Cocucci M (1997), Involvement of Ca+- calmodul in in Cd2+ toxicity during the early phases of radish (Raphanussativus L.) seed germination. Plant Cell Environ 20: 600–608.

Shaûq M, Iqbal MZ (2005), Tolerance of Peltophorumpterocarpum D. C. Baker Ex K. Heyne seedlings to lead and cadmium treatment. J New Seeds 7:83–94

Shanker AK, Cervantes C, Loza-Tavera H, Avudainayagam S (2005),Chromium toxicity in plants. Environ Int 31:739–751.

Sharma DC, Chaterjee C, Sharma CP (1995), Chromium accumulation and its effects on wheat (Triticumaestivum L. cv. DH220) metabolism. Plant Sci. 111:145–151.

Shukla OP, Dubey S, Rai UN (2007), Preferential accumulation of cadmium and chromium: Toxicity in Bacopamonnieri L. under mixed metal treatments. B Environ Contam Toxicol 78:252–257.

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Original Paper

ISSN: 2321-1520

133 Vidya (2019) Vol : 14(2) Chavda

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Original Paper

ISSN: 2321-1520

Himanshu v. Limbachiya

Ph.D. Research Scholar Education Dept, Gujarat University, Ahmedabad E-mail : boby [email protected]

* corresponding Author

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Review Article

ISSN: 2321-1520

Root Canal Irrigantsin Pediatric Dentistry: A Literature Review & Update

Vaibhav Kotecha*, Anupkumar Panda, Vishwa Y. Patel

Department of Pedodontics& Preventive Dentistry, College of Dental Sciences & Research Center, Ahmedabad E-mail – [email protected]

*Corresponding Author

Received Date:10-6-2019

Published Date: 15-9-2019

Abstract

Successful root canal treatment is dependent on the removal of microorganisms from the pulp and other anatomical irregularities of the root canal system. However, complete elimination of bacterial contaminants as well as necrotic debris require adjunctive use of root canal irrigants along with mechanical instrumentation. Irrigants can augment mechanical debridement by flushing out debris, dissolving tissue, and disinfecting the root canal system. Chemical debridement is especially needed for primary teeth with complex internal anatomy and zones inaccessible to debridement, such as accessory canals, ramifications, and dentinal tubules that might be missed by instrumentation. None of the available irrigating solutions alone provides all the ideal requirements. Studies have shown that a combination of two or more irrigating solutions in a specific sequence will help to achieve optimal irrigation. This review article mainly highlights the mechanism of action, safety and biocompatibility of currently used irrigation solutions and also the other materials that can be used

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as a potent irrigants, their advantages and limitations infuture.

Keywords:Root canal irrigants, primary tooth, smear layer, dentin permeability, antimicrobialproperties.

Introduction

The success of endodontic therapy in primary teeth strongly depends on achieving an adequate level of disinfection within their root canals5. Mechanical instrumentation alone is unlikely to be sufficient in attaining such disinfection, considering the root resorption process and the complex anatomy of the root canal system, characteristic of primary molars, and the risk of damage to the permanent germ6 7. Evidence has shown that the mechanical instrumentation technique with files is limited because it tends to leave significant portions of the infected canal walls untouched; thus, an important number of viable pathogenic microorganisms persist, lodged together with dentin debris and necrotic pulp-tissue remnants inside the dentin tubules, canal ramifications, and resorption craters8. Therefore, it is necessary to significantly reduce or to eradicate, to the extent possible, the microorganisms and their by-products present to the pulp canals by employing clinically effective and biocompatible irrigants, which also aid in dissolving organic debris9. Irrigation currently represents the best method in paediatricpulpectomy for the lubrication and flushing away of loose necrotic and contaminated materials during instrumentation10.

RATIONALE FOR USING ROOT CANAL IRRIGANTS

While various chemical and physical irritants can cause irritation and even necrosis of the pulp, the most common causes for pulpal inflammation (pulpitis) are bacteria and/or their products entering the pulp through a deep caries lesion or a leaking filling, e.g. an inflammatory reaction in the pulp starts long before bacteria invade the pulp tissue. The inflammatory reaction is first initiated by bacterial antigens interacting with the local immune system. Although no exact data are available, it is likely that the majority of bacteria inmost primary

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Sr Category Agents Ingredients Major No. Advantage(s)

1 Antiseptic Sodium 0.5-5.25% Tissue dissolution and/or Hypochlo available and Disinfectant rite chlorine antimicrobial Antimicrobial 2 Antiseptic 0.1-2.0% and/or Gluco Chlorhexidine Disinfectant nate Gluconate

3 Oxidizing Hydrog 3% Hydrogen Effervescence agents en Peroxide with NaOCl Peroxid (beneficial e effect in the canal questionable)

4 Oxidizing Urea 10% Urea Good wetting agents Peroxide Peroxide, ability, excellent (Endo- 15% lubricant PTC) Tween 80 and 75% Carbowax

5 Oxidizing Urea 10% Carbamide Good wetting agents Peroxide peroxide in ability, excellent (Gly- glycerol lubricant Oxide)

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6 Chelating Ethylenediam 10–17% Softens dentine agent ine recommended and removes (partially) root canal infections are located in the main root canal, while a minority of the cells would have invaded further into the dentinal tubules and lateral canals In root canal treatment, cleaning is the removal of all contents of root canal system before and during shaping. Irrigation is presently the best method for lubrication, destruction of microbes, the removal of tissue remnants, and dentin debris during instrumentation. The simple act of irrigation allows the flushes away loose, necrotic, contaminated materials before that they are inadvertently pushed deeper into the canal and apical tissues, compromising the periapical tissue and permanent bud. In this context, the use of cleansers in the irrigation process is essential.1

IDEAL REQUISITES OF A ROOT CANAL IRRIGANT

The ideal requisites of a root canal irrigant as given by Zehnderare4:

x Broad antimicrobialspectrum

x High efficacy against anaerobic and facultative microorganisms organized inbiofilms

x Ability to dissolve necrotic pulp tissueremnants

x Ability to inactivateendotoxin.

x Ability to prevent the formation of a smear layer during instrumentation or to dissolve the latter once it hasformed.

x Systemically nontoxic when they come in contact with vital tissues, noncausticto periodontal tissues, and with little potential to cause an anaphylacticreaction.

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TABLE 1: Root canals cleansers, which have been studied, for use in endodontic treatment.2

Tetra smear layer Acetic Acid

7 Chelating EDTAC EDTA with Good wetting agent ability for EDTAC preparations

8 Chelating RC-Prep EDTA and Urea Excellent agent Peroxide in a lubricant base of carbowax

9 Organic Acid Citric acid 10-50% Removes smear recommended layer

10 Organic Acid Polyacrylic 5-20% Removes smear acid recommended layer

11 Organic Acid Tannic acid 25% solution Removes smear layer

EFFICACY OF CLEANSERS In root canal treatment, cleaning is the removal all contents of root canal system before and during

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shaping. Irrigation is presently the best method for lubrication, destruction of microbes, the removal of tissue remnants, and dentin debris during instrumentation. The simple act of irrigation allows the flushes away loose, necrotic, contaminated materials before that they are inadvertently pushed deeper into the canal and apical tissues, compromising the periapical tissue and permanent bud. In this context, the use of cleansers in the irrigation process is essential12. Many researchers have studied the effect of several cleansers on the permeability of the dentine using methods that involve bacteria or radioisotopes, with different methodologies. Those cleansers have been used with the objective of eliminating pulpal remains and residues. In addition, they increase the dentin permeability (removing the smear layer), facilitate the instrumentation and promote the cleaning and disinfection of the root canals13-21. In addition, they should be soluble in water and biocompatible to the periapicaltissues22. VARIOUS ROOT CANAL IRRIGATING SOLUTIONS 1. NORMAL SALINE4 x Normal saline is isotonic to the body fluids. It is universally accepted as the most common irrigating solution in all endodontic and surgical procedures. It is also found to have no side effects, even if pushed into the periapical tissues. However, saline should not be the only solution to be used as an irrigant, it is preferably used in combination with or used in between irrigations with other solutions like sodiumhypochlorite. 2. SODIUM HYPOCHLORITE(NaOCl)1 x Henry Drysdale Dakin and the surgeon Alexis Carrel popularized the use of buffered 0.5% sodium hypochlorite solution for the irrigation of infectedwounds. x Sodium hypochlorite is a weak alkali that acts on the albumin (remains of pulpal tissue, foods and microorganisms), denaturing them and turning them soluble in water. It acts on microbial cells disrupting their vital functions leading to celldeath. x The most effective irrigation regimen is reported to be 5.25% at 40 minutes. Irrigation using 1.3% and 2.5% sodium hypochlorite for this same time interval is ineffective in removing E faecalisfrom infecteddentin. x Altering the pH, temperature and use of ultrasonic agitation increases the efficacy of sodium hypochlorite. Pure hypochlorite solutions, as are used in endodontics, have a pH of12. x Bloomfield and Miles confirmed that hypochlorites possess greater antimicrobial activity

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at a lower pH. x Rising temperature by 25 °C increases efficacy by a factor of100. x The dissolution power of 1% sodium hypochlorite at 45 °C is equivalent to that of a 5.25% solution at 20°C. x Sonic irrigant activation and ultrasonic irrigation with a nickel-titanium tip increases the effectiveness of 5% sodium hypochlorite in the apical third of the rootcanal. x A study evaluating effects of sodium hypochlorite on nickel-titanium lightspeed instruments concluded that no significant corrosion of nickel-titanium files in these solutions was detected. Sodium hypochlorite irrigation leads to decreased bond strength between dentin and resin cements. Certain reversal agents like ascorbic acid or sodium ascorbate reverse this reduction in bond strength completely. x The antimicrobial effect of 2.5% sodium hypochlorite used in combination with 0.2% chlorhexidine was greater than that of either agent used separately. But a carcinogenic product parachloroanaline (PCA) is produced by the reaction between sodium hypochlorite and chlorhexidine and the possibility of its leakage into the surrounding tissues is aconcern. x Though sodium hypochlorite is widely used as an irrigant in endodontics, it has an unpleasant taste and odour, removes the smear layer only partially, does not consistently disinfect the root canal, is toxic to the periradicular tissues and damages the permanent tooth follicle, peripheral tissues and oralmucosa. 3. CHLORHEXIDINE GLUCONATE2 x ChlorhexidineGluconate, currently used in endodontic therapy, seems to act by adsorbing onto the cell wall of the microorganisms and causing leakage of the intracellular components. At low concentrations, small molecular weight substances will leak out, especially potassium and phosphorus, resulting in a bacteriostatic effect. At high concentrations, chlorhexidinegluconate has a bactericidal effect due to the precipitation and/or coagulation of the cellular cytoplasm, probably caused by cross-linkingproteins. x Vahdaty et al. evaluated in vitro the antibacterial efficiency of 2% and 0.2% chlorhexidine, comparing them with sodium hypochlorite in the same concentrations. These cleansers were used in the infected dentin tubules. The results indicated that both substances reduced the number of bacteria in the superficial layers of the

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dentintubules. x Heling and Chandler and White et al. suggested that chlorhexidine can be an excellent antimicrobial endodontic irrigating agent if used alone, or as an auxiliary to sodium hypochlorite during the instrumentation. ChlorhexidineGluconate showed quick residual antimicrobial activity in these in vitrostudies. x Gomes et al. evaluated the antimicrobial activity of the two formulations of ChlorhexidineGluconate (liquid and gel) in three concentrations (0.2%, 1.0% and 2%), and of sodium hypochlorite (0.5%, 1.0%, 2.5%, 4.0%). The results showed that chlorhexidine in liquid form eliminated bacterial cells more quickly than the chlorhexidine gel. Even though all tested cleansers possessed antimicrobial activity, the time required to eliminate the studied microorganisms depended on the concentration and of the type of cleansersused. x Ferraz et al. evaluated 2% chlorhexidinegluconate gel as an endodontic irrigating agent according to its capacity to disinfect root canals contaminated with Enterococcus faecalis. Furthermore, they tested chlorhexidine gel cleaning capacity when compared with 5.25% sodium hypochlorite and 2% chlorhexidinegluconate solution. The results demonstrated that the chlorhexidine gel produced cleaning of the surface of the root canal and presented antimicrobial capacity comparable with the other appraised solutions. It could be concluded that chlorhexidine gel has satisfactory potential to be used as an endodontic irrigating agent. Most of the studies have been undertaken on permanent teeth in vitro, demonstrating the properties of the cleanser used for the instrumentation of those canals. This literature review found only two studies regarding primary teeth and root dentin permeability. x Bengtson et al. concluded that urea peroxide/Dakin’s liquid showed the highest dye penetration index of primary root dentin permeability. Primo verified that 1% sodium hypochlorite associated with 10% citric acid was the most effective association to remove the smear layer of anterior primary teeth, followed by the associations urea peroxide/Dakin´s liquid and 4% sodium hypochlorite /3% hydrogen peroxide. All of these solutions produced an increase in the primary dentin permeability. Therefore, other researches should be undertaken to indicate an effective substance for the chemical- mechanic preparation of primary teeth, as well as to verify the physiochemical properties

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of treatment solutions, providing greater sanitation and appropriate preparation of the rootcanals. 4. ETHYLENEDIAMINE TETRA ACETIC ACID(EDTA)2 x Ethylenediamine Tetra Acetic Acid is a chelating substance capable of removing calcium ions of the dentin, giving rise to demineralization and as a consequence, increasing the dentin permeability of the root canals. Ethylenediamine Tetra Acetic Acid is used in concentrations from 10 to 17% and in association with other drugs. The efficiency of chelating agents generally depends on many factors, such as root canal length, penetration depth of the material, hardness of the dentin, application time, pH, andconcentration. Nakashima and Terataobserved that the permeability of root canal disinfectants increased to similar degrees in the 3% and the 15% ethylenediamine tetra acetic acid groups. Comparing dentin properties, they propose that 3% ethylenediamine tetra acetic acid is more useful for clinical applications. x Zuolo et al. found that the most effective combination to increase root dentin permeability was ethylenediamine tetra acetic acid associated with Cetavlon(EDTAC). x Tao et al. verified that ethylenediamine tetra acetic acid did not modify the root dentin permeability. They suggest that the absence of changes in the root dentin permeability with a conventional endodontic preparation was due to the fact that, even though endodontic preparation reduces dentin thickness, it also created a smear layer that compensated to the extent that there was no overall change in permeability. A new chelating agent (Glyde File Prep) containing ethylenediamine tetra acetic acid has been proposed for permanentteeth. x Grandini et al. evaluated the smear layer, debris, and tubule orifices of root canal walls after being instrumented and irrigated by Glyde File Prep, using a scanning electron microscope. The results of this study confirm that irrigation with sodium hypochlorite alone was not able to remove totally the smear layer, because its action is mainly directed to the organic debris. To obtain the total removal of the smear layer, that is, both organic and inorganic components, the combined use of sodium hypochlorite and ethylenediamine tetra acetic acid is recommended. The chelating agent prepares the canal wall surfaces so that cleansers and medications are effective with their antibacterial action. One of the most effective drugs for removing the smear layer is RC-Prep

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(ethylenediamine tetra acetic acid /Urea Peroxide). It is a potent bactericidal agent and increases the dentin permeability significantly. x Others substances (organic acids) have been used to remove the smear layer, such as 6 - 10% citric acid, 20% polyacrylic acid, and tannic acid. Salamaand Abdelmegid found that irrigation with 6% citric acid for 15 or 30 seconds was effective in removing all smear layer components of the primary root canals. However, further researches are needed to investigate the biocompatibility of acids and to test combinations of solutions. Moreover, these acids could have a harmful effect on the periapical tissues of both permanent and primaryteeth. 5. UREA PEROXIDE2 x Another widely used solution to aid instrumentation is Urea Peroxide (Endo-PTC or Gly-Oxide). The peroxides are oxidizing agents that react chemically, liberating great amounts of nascent oxygen that explains their bactericidal action. The effervescence, due to the liberation of oxygen, contributes to the removal of pulp tissue remains and dentin particles during the chemical-mechanicpreparation. x In Brazil, the trade name of Urea Peroxide is Endo-PTC (10% Urea Peroxide, 15% Tween 80 and 75%Carbowax). x International literature points out that Urea Peroxide is marked as Gly-Oxide commercial brand. It is anhydrous glycerol based, without any added detergent. x Moura and Paivausing Endo-PTC as an auxiliary chemical substance, observed less dye penetration with instrumentation increasing, mainly in the apical area. The Urea Peroxide hasseveraldesirable characteristics for the irrigation of root canals in primary teeth. It presents detergent and haemostatic properties, besides not being irritating to the periapical tissues and non allergenic. x Stewart et al. and Rome et al. observed that the bactericidal activity of the Urea Peroxide (Gly- Oxide) was superior to 3% Hydrogen Peroxide in the preparation of infected root canals. The association of Urea Peroxide/sodium hypochlorite maintains the previously describedproperties. x According to Rome et al., the use of Urea Peroxide is the first choice cleanser in small curved canals. Its properties of lubrication without demineralization the dentin walls avoid the risks of root perforation, common in primary teeth. The association of Urea

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Peroxide with sodium hypochlorite promotes significant more increase in the dentin permeability index to dye and drugs than when used separately. In spite of promoting increase in the dentin permeability, the association of Urea Peroxide/sodium hypochlorite showed less effectiveness in removing the smear layer. In contrast, it is known that the smear layer reduces dentin permeability, and prevents the penetration of root canal disinfectants into the deep area of the root canalwall. 6. MIXTURE OF TETRACYCLINE ISOMER, ACID, AND DETERGENT(MTAD)3 x Biopuremtad has been introduced to dentistry as a final irrigantfor smear layer removal. Mtad has been proved to be effective in eliminating resistant microorganisms and providing sustained antimicrobial activity. Minimal erosion of intra-radicular dentin has been reported after final canal irrigation withmtad. x Torabinejadet al.developed airrigantwith combined chelating and antibacterial properties. Mtad (a mixture of tetracycline isomer, acid, and detergent, biopure, tulsadentsply, tulsa ok, usa) is a new product in the quest for a better root canal irrigant, with a phas low as 2.15. In their study, the above authors used this new irrigant, focusing on the removal of smear layer, extracted single-rooted teeth were prepared by using passive stepback and rotary 0.04 taper niti files. Distilled water or 5.25% sodium hypochlorite was used for irrigation followed by 5ml irrigation with one of the following: sterile distilled water, 5.25% sodium hypochlorite, 17% ethylenediamine tetra acetic acid, or mixture of tetracycline isomer, acid, and detergent. The results indicated that mtad is an effective solution for the removal of the smear layer and does not significantly change the structure of the dentinal tubules, when canals are first irrigated with sodium hypochlorite, followed by a final rinse of mixture of tetracycline isomer, acid, and detergent. Mixture of tetracycline isomer, acid, and detergent is an irrigant solution with ingredients capable of disinfecting the dentin, removing the smear layer, opening the dentinal tubules and allowing the antibacterial agents to penetrate the entire root canal system. x In another study, the same group investigated the effect of various concentrations of sodium hypochlorite as an intracanalirrigant before irrigation with mixture of tetracycline isomer, acid, and detergent as a final rinse on the smear layer. The results showed that mtad removed most of the smearlayer when used alone; however, remnants of the

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organic component of the smear layer could be detected on the root canal walls. There were no significant differences between the ability of 1.3%, 2.6%, and 5.25% sodium hypochlorite as root canal irrigants and mtad as a final rinse to remove the smearlayer. 7. CITRIC ACID3 x Citric acid can also be used for irrigation of the root canal to remove the smear layer. Concentrations ranging from 1% to 50% have been used. The use of 10% citric acid as final irrigation has shown good results in smear layer removal and proven to be more biocompatible than 17% EDTA-T and 17% ethylenediamine tetra aceticacid. x Gutmann et al showed that 10% citric acid was more effective in removing the smear layer from apical root-end cavities thanultrasound. x Yamaguchi et al compared the chelating and antibacterial properties of citric acid andethylenediamine tetra acetic acid. Powdered dentine–resin mixture was found to be more soluble in a 0.5, 1, and 2M citric acid solutions than in a 0.5M ethylenediamine tetra acetic acid solution. Citric acid solution showed antibacterial effects on all 12 root canal bacteriatested. x Di Lenarda et al and Scelza et al reported a minor or no difference in smear layer removal with citric acid and 15% ethylenediamine tetra acetic acid. The use of 25% citric acid was found to be ineffective in eradication of biofilms of E faecalisafter 1, 5, and 10 minsofexposure. x Machado-Silveiro et al measured the demineralization capability of 1% and 10% citric acid, 10% sodium citrate and 17% ethylenediamine tetra acetic acid during immersions of 5, 10 and 15 min on root canal dentine. Ten percent citric acid was more effective than 1% citric acid, which was more effective than ethylenediamine tetra aceticacid. x Hariharan et al conducted an in vitro study to determine the efficacy of 5.25% sodium hypochlorite, 5.25 sodium hypochlorite + 10% ethylenediamine tetra acetic acid, 6% citric acid and 2% chlorhexidine and saline (control) in removing the smear layer in primary teeth root canals after hand instrumentation. They showed the superior efficacy of 6% citric acid than the other tested irrigantson removing the smear layer in primary teeth. Both ethylenediamine tetra acetic acid and citric acid can effectively remove the smear layer created during canal instrumentation. Although citric acid may also have an antibacterial effect, this has not been compared with other root canal disinfecting agents

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in in vitro or in vivostudies. 8. OZONATED WATER4 x Ozone is a chemical compound consisting of three oxygen atoms (O3-triatomic oxygen), which has higher energetic form than normal atmospheric oxygen. Ozone is capable of oxidizing any biological entity due to its powerful bactericidal properties where even at low concentration, (0.1 ppm), it is capable of inactivating bacterial cells including their spores. Although it is a powerful antimicrobial agent, less attention has been paid to its antibacterial activity on bacterial biofilm and hence in root canalinfection. x Studies have found that when the specimen was irrigated with sonication killing ability of ozonated water and 2.5% sodium hypochlorite was almost comparable. However, it was found that effecton E. faecaliswas more with sodium hypochlorite than ozonated water in broth culture and in biofilm. Hence concluded that there is need for further studies and modifications in ozonated water before it could be used as a root canal irrigant. Photon activated disinfection (PAD): Photon activated disinfection is based on the concept where light of the appropriate wavelength is used to preferentially localize the nontoxic photosensitizers to generate singlet oxygen and free radicals that are cytotoxic to cells of the target tissue. Blue dyes, especially toluidine blue and methylene blue used with a 632.8 nm wavelength laser have shown significant result. A study in primary teeth showed a microbial reduction of 98.37% after using photodynamic therapy. Thus in future photon activated disinfection could be a low-cost and nontraumatic alternative to other irrigants to be used in today’s treatment of primaryteeth. 9. CARISOLV4 x Carisolv contains 0.5% sodium hypochlorite along with amino acids. Thus it was hypothesized that this agent can also be effective in removal of smear layer in root canal when used as anirrigant. x The mode of action is to degrade the denaturedcollagen. x A study which compared carisolv with 17% ethylene diaminetetraacetic acid and 5.25% sodium hypochlorite in removing layer on radicular dentin showed that carisolv was ineffective in removal of smear layer. The reason attributed was because it was in gel form which made it difficult to wet and flush thecanals.

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x A study was done to compare the efficacy of carisolv, 1% sodium hypochlorite gel, and 1% sodium hypochlorite solution as root canal irrigants in primary teeth showed that carisolv was better compared to sodium hypochlorite gel in cleaning the debris at the apical third and hence concluded that carisolv can be used as an effective root canalirrigant. 10. TETRACLEAN4 x Tetraclean is a mixture of an antibiotic, an acid (citric acid), and a detergent (polypropylene glycol). The concentration of citric acid and the type of detergent used varies from that of a mixture of tetracycline isomer, acid, and detergent. The properties of tetraclean are due to its low surface tension which enables better adaptation of the mixtures to the dentinal walls. It removes the smear layer, effective against strictly anaerobic and facultative anaerobic bacteria like E.faecalis. 11. ELECTROCHEMICALLY ACTIVATED SOLUTIONS4 x A mixture of tap water and low concentrated salt solutions forms the electrochemically activated (ECA) which results in synthesis of Anolyte, and Catholyte. The oxydizing substances of anolyte exhibits microbicidal activity against bacteria, viruses, fungi and protozoa and therefore termed Superoxidized Water or Oxidative Potential Water. Due to its various advantages like ease of removal of debris and smear layer even at the apical third as well as its nontoxic properties it can be used as a potent root canalirrigant. x A study showed that Oxidative Potential Water was as effective as the sodium hypochlorite when used as an irrigant in necrotic pulpectomized primary teeth and is suggested as an alternative for irrigating primaryteeth. 12. SMEAR CLEAR1 x Smear Clear is a recently introduced chelating agent that contains 17% ethylenediamine tetra acetic acid solution with two additional proprietarysurfactants. x Smear Clear contains cetrimide, which is a quaternary ammonium compound and a cationic detergent that is effective against gram positive and gram negative microorganisms, hence it has been found to be effective againstE.fecalis. x All the studies that have evaluated this product have used it for 1 minute as per according to the manufacturer'sinstructions.

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13. QMIX1 x QMix 2 in 1 solution is a newly introduced irrigation solution developed and marketed by Dentsply Tulsa Dental Specialties, Tulsa, OK, USA. x It contains a mixture of a bisbiguanide antimicrobial agent chlorhexidine, a polyaminocarboxylic acid calcium-chelating agent ethylene diaminetetraacetic acid, and a surfactant cetrimide mixed in distilled water with acceptable additional salt. It is recommended as final irrigant during root canal procedures. x It eradicates bacteria, removes smear layer, and persists in biofilms. x It has pH slightly aboveneutral. 14. SILVER DIAMINEFLUORIDE1 x A 3.8% w/v silver diamine fluoride solution has been used for intracanal irrigation. Silver diamine fluoride has potential for use as an antimicrobial root canal irrigantor interappointment medicament to reduce bacterial loads. 60 minutes exposure to silver diamine fluoride completely killed Efaecalis, but silver particles occluded dentinal tubular orifices after removal of the smearlayer. 15. HEBP (1-HYDROXYETHYLIDENE- 1, 1-BISPHOSPHONATE)1 x Also known as etidronic acid or etidronate. It is a potential alternative to ethylene diaminetetra acetic acid or citric acid because this shows no short-term reactivity with sodiumhypochlorite. 16. TRICLOSAN AND GANTREZ1 x Triclosan is a broad spectrum antimicrobial agent, which acts against gram-positive and gram- negative bacteria as well as some fungi andviruses. x The addition of Gantrez enhanced the bactericidal activity oftriclosan. x Triclosan alone and in combination with Gantrez demonstrated bactericidal activity against the five specific endodontic pathogens P intermedia, F nucleatum, Anaeslundii, P gingivalis, and Efaecalis. 17. ENDOVAC SYSTEM1 x The EndoVac system is a new irrigation system that consists of a delivery/evacuation tip attached to asyringeofirrigantandthe high-speed suctions ource of the dental unit. As the can nulasa replaced in the canal, negative pressure pulls irrigantfrom a fresh supply in the chamber down into the canal to the tip of the cannula, then into the cannula and finally out through the suctionhose.

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18. HERBAL IRRIGANTS4 x The use of herbal products in the field of medicine has been practiced since ancient times and has significantly increased over the last fewdecades. x In recent endodontics because of the limitations of most of the commercial intracanal medicaments used such as cytotoxicity and their inability to eliminate bacteria from dentinal tubules, trend of recent medicine to use biologic medication extracted from natural plants is drawing a lot ofattention. x The major advantages of using herbal alternatives are easy availability, cost-effectiveness, increased shelf life, low toxicity, and lack of microbial resistancereported. x Literature has shown that herbs can have a promising role as root canalirrigants. 19. MISWAK4 x Miswak is derived from the plant Salvadorapersica(known asÿarakÿ in Arabic) mainly used as a chewing stick, which is used for cleansing theteeth. x Wolinskyand Sote/, by isolation of the active ingredient from S. persica, found that the limonoid had a great antimicrobial activity by inhibiting the growth of various Gram- positive and Gram- negative microorganisms by interfering extrapolysaccharides and glycosidase enzymes produced by thesemicroorganisms. x An in vitro study showed that 10 to 20% miswak extract was effective antifungal and antibacterial agent when used as an irrigant in the endodontic treatment of teeth with necrotic pulps against candida albicansand E.fecalis. x A study done on primary teeth showed that for root canal irrigation miswak could be a good natural substitute to sodiumhypochlorite. 20. MORINDA CITRIFOLIA4 x Morindacitrifoliacommercially known as Noni or India Mulberry, has a broad range of therapeutic effects such as antibacterial anti-inflammatory, antiviral, antitumor, anthelmintic, analgesic, hypotensive, anti-inflammatory and immune enhancing effects might be advantageous. Due to its properties and not likely to cause the severe injuries to patients that might occur through sodium hypochlorite accidents its juice can be used as a potent irrigant in primary teeth. The antimicrobial activity on E. faecalisof 2% chlorhexidine gel, propolis, M. citrifoliajuice and calcium hydroxide has beencompared.

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x It was concluded that Propolis and M. citrifoliawere effective against E. faecalisin dentin on extractedteeth. 21. TRIPHALA AND GREEN TEA PHENOLS4 x Triphala is one of the well knownIndian Ayurvedic herbal formulation consisting of dried and powdered fruits of three medicinal plants namely Terminal iabellirica(Gaertn) Roxb, Terminal iach ebula Retz and EmblicaofficinalisGaertn. Green tea polyphenols- Numerous human, animal and in vitro studies have shown anticariogenic, anti- inflammatory, thermogenic, probiotic and antimicrobial properties. x An in vitro study showed maximum antibacterial activity with sodium hypochlorite and statistically significant antibacterial activity with Triphala, GTPs, a mixture of tetracycline isomer, acid, and detergent and 5% hypochlorite against E. faecalisbiofilm. 22. GERMAN CHAMOMILE AND TEA TREEOIL4 x German chamomile is a medicinal plant known for its anti-inflammatory, analgesic, anti- microbial, antispasmic and sedative properties. Chamomile was found to be effective when used as a mouthwash and many properties such as antiseptic, antifungal agent and a mildsolvent. x An scanning electron microscopic study done on German chamomile extract and tea tree oil as irrigants and showed that efficacy of chamomile to remove smear layer was superior to sodium hypochlorite alone but less than sodium hypochlorite combined with ethylene diaminetetraacetic acid. 23. PROPOLIS4 x Propolis, a resinous beehive product is a potent antimicrobial, antioxidant and anti- inflammatory agent. Very few studies have evaluated propolis as a root canal irrigant. Comparative evaluation of antimicrobial activity of miswak, propolis, sodium hypochlorite and saline as root canal irrigants by microbial culturing and quantification in chronically exposed primary teeth was done showed that results of propolis was comparable to those of the negativecontrol. x An in vivo randomized trial was conducted where root canals were irrigated using either 2% chlorhexidine, 4% calcium hydroxide extract of propoliswith normal saline as the control irrigant which showed a significant decrease in mean aerobic colony forming

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units count in all the groups, however chlorhexidine was superior. Propolis has also shown to be an effective intracanalirrigant in eradicating E. faecalisand C.albicans. INFLUENCE OF IRRIGATION METHODS2 x A variety of instrumentation and irrigation methods have been used in endodontic treatment. According to literature researched, the instrumentation and the root canals irrigation can be accomplished via manual conventional means (endodontic files and Luer syringes) or via endodontic ultrasonic-vibration- generator systems. The endodontic preparations might induce changes in the root dentin permeability. When a file is ultrasonically activated and placed passively in a canal, a phenomenon called acoustic streaming is produced, which is one of the surported mechanisms for superior debridement. Biological material that enters the streaming fields would be subjected to large shear stresses and may be disrupted. Ultrasonically prepared teeth showed cleaner canals than the teeth prepared by handinstrumentation. x Regarding primary teeth, Seowconcluded that a combination of mechanical filling followed by ultrasonication produced the best results, with 95% of bacteria removed. The results showed that the ultrasonication might be useful for primary teeth endodontic treatment. However, the present literature search has found studies, which were carried out on teeth whose instrumentation was accomplished with endodontic files and they were irrigated with Luersyringes (sodiumhypochlorite). x Hata et al. affirmed that the manual irrigation technique in primary teeth was more effective to remove the smear layer using 15% ethylene diaminetetraacetic acid irrigation associated with 5% sodium hypochlorite during instrumentation. However, the most effective irrigation technique to remove debris was the ultrasonic system, regardless of the cleanserused. x Cunningham and Martin observed a good rate of smear layer removal from root canals irrigated with 2.5% sodium hypochlorite, using the Endosonicsystem. Nevertheless, there were no significant differences among the manual and ultrasonic instrumentation techniques to reduce the smear layer effectively in permanent teeth, and consequently no permeability alterations were found. The variation of hydraulic conductance measured in situ after three endodontic preparations techniques (manual, ultrasonic, and manual with sodium hypochlorite and ethylene diaminetetraacetic acid) showed an inverse

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relationship between variations in dentin permeability and the presence of smear layer. Dentin thickness was a significant factor influencing radicular permeability as well as the smear layer. The use of ethylene diaminetetraacetic acid induced a considerable increase in radicular permeability and the use of ultrasonics produced a similar but weakereffect. x Cameron affirmed that the most effective method to increase the root dentin permeability, considering removal of smear layer, was a manual instrumentation (endodontic files), irrigation with EDTAC (Ethylene diaminetetraacetic acid associated to Cetavlon), followed by the use of ultrasound with EDTAC for 1 minute and ultrasound with 4% sodium hypochlorite for 2minutes. x Further researches should be carried out to evaluate the best irrigation method for primary teeth, which should provide increase of root dentinpermeability. x Based on literature reviewed, it can be concluded that among the solutions used for permanent teeth, all solutions and associations studied increased the dentin permeability. There was no difference in the dentin permeability when comparing manual and ultrasonic irrigation. There are not enough papers focusing primary teeth on this subject to establish the patterns of increased dentin permeability in tooth root canal treatment regarding the irrigating solutions and irrigationsystems. SEQUENCE OF IRRIGATION23 During the last several years endodontics has progressed to the point for treatment to be less traumatic for the patient and less stressful for the dentist. However, if the use of nickel titanium rotary instruments has allowed us to gain time during endodontic treatment, it can also tempt us to neglect one of the main objectives of endodontics, that being the "cleaning" on which Dr. Herbert Schilder has emphasized since he insisted on "cleaning and shaping". It may be more appropriate to say, "shaping for cleaning". The main goal of the root canal treatment is to completely eliminate the different components of the pulpal tissue, calcification and bacteria, the placement of a hermetic seal to prevent infection or re-infection and to promote healing of the surrounding tissues if needed. Irrigation has an important role during endodontic treatment. We must ask ourselves the question, "Why do we irrigate and what irrigation protocol will provide the cleanest canal?" In this context, let us remember that the shaping is the result of endodontic instruments while the cleaning results from irrigation. Therefore, we have two types of preparation. The first one is chemical and the

162 Vidya (2019) Vol : 14(2) Kotecha et al second one is mechanical. Moreover, it has been proven that there is a close correlation between these two types of preparation. In fact, with greater tapered preparations, the quantity and the concentration of the irrigating solution will be greater and will therefore better eliminate the smear layer. The files can clean only parts of the root canal system. They create a reservoir that can hold the different irrigating solutions, which will access and clean portions of the root canal system that the instruments cannotreach. Here’s a sequence for the different irrigating agents in order to achieve the best chemical preparation possible: VITAL TEETH:In this case there’s a challenge of treating the complexity of the different components of the pulp, and eventually the presence of bacteria. Begin the treatment by: 1) An application of sodium hypochlorite and/or an application of urea peroxide. The purpose of this mixture: a. The collagenic anti-aggregation effect due to the proteolytic and lipidic affinity of urea peroxide. b. To destroy the biggest amount of pulp tissue inside the access cavity and provide a better view of the canal orifices by controlling bleeding and preventing any collagenic plugs fromforming. c. At this stage the effect of ethylene diaminetetraacetic acid is only important for its antibacterial effect in combination with other antibacterialagents. 2) The second step consists of irrigating with 2ml of sodium hypochlorite 5.25 percent (60°C). The warm sodium hypochlorite is more efficient in destroying the collagen and this will reduce the time needed for the elimination of the organic part. This irrigation will create an effervescent effect between the sodium hypochlorite and urea peroxide. This "elevator effect" will evacuate the organic debris outside the access cavity, disorganize the coronal pulp tissue and help to better detect the canalorifices. 3) A second application and its activation is obtained by using a K file (08-10). This will disorganize the pulpal tissue in both the cervical and middle thirds of the endodontic system.This step has to be preceded by an abundant irrigation with distilled water in order to eliminate the first mixture present in the accesscavity. 4) Once the preparation of the canal has begun, Smear Clear (Sybron Endo, Orange, CA) (17 % ethylene diaminetetraacetic acid, cetrimide and surfactants) must be used.

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The ethylene diaminetetraacetic acid is an organic acid which eliminates the mineral part of pulp tissue, the surface tension inhibitor will allow a better contact with the dentin for a higher efficiency of the product. It is advised to alternate the use of ethylene diaminetetraacetic acid from the beginning of the preparation in order to eliminate the mineral layer before its thickening and condensing it inside the canal systems which will close the entrances of lateral and accessory canals and dentinaltubules. Each time a rotary file is working inside the canal, irrigating solution must be present. Ultrasonic activation of the irrigating solution, using a small diameter file, is advised for a more efficient chemical preparation. The early use of ethylene diaminetetraacetic acid facilitates the flow of the different irrigants in the lateral canals permitting a chemical preparation of all the endodontic system. Ethylene diaminetetraaceticacid also plays an important role in the reduction of inflammatory reaction by inhibiting the affinity of macrophages to the vaso-active peptides of the pulpal tissue. A time frame between four and five minutes must not be exceeded for the presence of the ethylene diaminetetraacetic acid inside thecanal. Chlorehexidine can be used for a total elimination of the bacteria inside the canal. Distilled water is used between each irrigating solution in order to prevent an acid/ base reaction, between sodium hypochlorite and ethylene diaminetetraacetic acid, for a more efficient action of the chemicals on the tissues. A copious neutralization of all the chemical agents must be done by the end of the preparation and before the fitting of the guttapercha cones so that the master cone does not push any of the chemicals outside the canal that might cause an inflammation of the surroundingtissues. NECROTIC TEETH: The main difference between vital teeth and necrotic ones is the absence, not in total, of the pulpal parenchyme and the abundance of bacteria present in the latter. For this reason, the irrigation sequence will be different. Irrigation will be initiated with either sodium hypochlorite (5.25%, 60%C) for its bacterial effect or with chlorohexidine (0.2%) for the elimination of various bacterial types present in the root canals and dentinal tubuli. Then using distilled water to neutralize the effect of these irrigants followed by which we can repeat the same irrigation sequence described previously for vital teeth. The ethylene diaminetetraacetic acid, by eliminating the smear layer and opening the dentinal tubuli will permit an easy flow of sodium hypochlorite or chlorhexidine for a better disinfection of the endodontic system. In both clinical situations (vital and necrotic

164 Vidya (2019) Vol : 14(2) Kotecha et al teeth) it is necessary to end our sequence by using distilled water in order to eliminate the chemical agents or to neutralize their effects. This will inhibit: - Their flow towards the periodontaltissues, - The alteration of the fillingmaterial, - The formation of a precipitating layer due to the crystallization of sodium hypochlorite after drying the canalwalls. PRESENCE OF RESORPTIONS:When there’s an internal resorption, the irrigation sequence is the same that was described for vital teeth. But this sequence will be followed by the use of citric acid 50 percent (10 minutes) in order to eliminate the granulation tissue and to obtain smooth dentinal walls. This will ameliorate the adaptation of the filling material. The citric acid is eliminated by sodium hypochlorite and distilled water. The same sequence is adopted for external apical resorptions but with an activation of the patency. Discussion The preservation of primary teeth is one the most important aims of paediatric dentistry. The aim is to maintain the harmonious growth and development of arch length and occlusal balance, with optimal function for swallowing, chewing, speech, and aesthetics10. Therefore, pulp therapies should be performed with high levels of quality to ensure a maximally successful clinical result. The combination of chemical and mechanical preparation is the key requisite for the achievement of the best eradication of debris. Irrigation has to be constant and copious to eliminate living/non-living materials. Currently, there is no agreement among paediatric dentists concerning the best intracanalirrigant solution for use against pulp pathogens involved in irreversibly inflamed/infected or non-vital primary teeth; this absence of consensus is probably due to the lack of researchbased evidence to support the use of any ‘best’agent8. The time we gain by using rotary Nickel-titanium instruments is compensated by an abundant irrigation for a better cleaning of the endodontic system and this will contribute to the increase success rate of endodontic treatment. Chemical preparation is a double-edged sword; it will help us succeed in the adequate cleaning of the main canal its systems. But it must be followed by a three dimensional obturation to fill all of what has been cleansed and prepared. Perfect absorption of the fluid is essential from the main canal and all of its systems. If this is not accomplished then the adherence between the sealer and the dentin will be compromised. In addition, the presence of the

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fluid inside the systems can have a negative hydraulic pressure preventing the obturation material from entering the complexity of the root canal systems for accomplishing a three dimensional obturation.23 Conclusion Successful root canal therapy depends on thorough chemomechanical debridement of root canal. Irrigants can augment mechanical debridement by rinsing out debris, dissolving tissue and disinfecting the root canal system. Significance of chemical debridement increases for teeth with complex internal anatomy such as tortuous canals or other irregularities that might be missed by mechanical instrumentation. Root canal instrumentation produces a smear layer consisting of organic and inorganic components that cover the root canalsurfaces. Irrigation plays an important role in successful debridement and disinfection. Most commonly used root canal irrigants are normal saline and sodium hypochlorite. Physiological saline has no effect on removing dentinal debris and smear layer and sodium hypochlorite is not effective in removing the smear layer and can be harmful to the peripheral tissues. Certain authors have reported that during instrumentation, canals should be irrigated utilizing copious amounts of sodium hypochlorite, and once the shaping procedure is completed, ethylene diaminetetraacetic acid or citric acid should be employed; these authors also mentioned that chlorhexidine or mixture of tetracycline isomer, acid, and detergent are strongly recommended as best final irrigant agent prior to canal drying and filling11. Currently, there is no agreement among paediatric dentists concerning the best intracanal irrigant solution for use against pulp pathogens involved in irreversibly inflamed/infected or non-vital primary teeth; this absence of consensus is probably due to the lack of researchbased evidence to support the use of any ‘best’ agent11. Researches are still under way on to scientifically validate an irrigant solution with ideal characteristics. REFERENCES 1. Ismail S, Adyanthaya A and Sreelakshmi N. Intracanal irrigants in pediatric endodontics: A review. International Journal of Applied Dental Sciences 2017;3(4):246-51. 2. Pascon FM, Kantovitz KR, Rontani RMP. Influence of cleansers and irrigation methods on primary and permanent root dentin permeability: a literature review. Braz J Oral Sci. July- September 2006;5(18).

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3. Nizami SK, Chaudhary P, Lodhi R, Syed M, Sharma MN and Thukral H. Irrigating solutions in pediatric dentistry – a review. World Journal of Pharmacy and Pharmaceutical Sciences;7(3):357-68.

4. Ramachandra JA, Nihal NK, Nagarathna C, Vora MS. Root Canal Irrigants in Primary Teeth. World J Dent 2015;6(3):229-34.

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