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Part II • Culturing of algae • Collection of algae

• Identification of algae

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Chapter 3. Collection, Culturing and Identification of algae.

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3.1. Introduction

Algae are naturally abundant, mostly autotrophic, found in all kinds of aquatic bodies,

with different environmental conditions. India has a coastline of 7500 Kms and also

plenty of fresh water bodies are available. Algae belonging to different divisions

thrive in these water bodies, thus making algal diversity an extremely large pool of

readily available and inexpensive biomaterials (Rao and Mantri, 2006). The marine

macroalgal resources along the Maharashtra coast have been studied since the

beginning of this century (Boergesen, 1931). In the present investigation, collection

of algae (for their possible use as biosorbents) along the sea coasts of Maharashtra,

and, from the estuarine and fresh water bodies was carried out. The Geographical

location of the place of research (University of Pune, Pune and BARC, Trombay) is

in the proximity of Western Ghats and adjacent to the west coast of India

respectively. In addition to the 760 Kms coast line of Maharashtra; plenty of brackish

and fresh water habitats are available. Large diversity of algae thrives in these

habitats. Algae are classified and identified on the basis of cell organization, type of

chlorophylls, cell wall chemistry, reserve food material, and flagellation. For

example, presence of chlorophyll a is a common characteristic in all algae; however,

the presence of other phytopigments is a characteristic of a particular division of alga

(Hoek et al., 1995)).

The importance of a biological system as a biosorbent for a metal depends on factors

like; the number of binding sites present on biosorbent, the accessibility of the sites,

the chemical state of the site (i.e. availability), and affinity between site and metal

Chapter 3. Collection, culturing, and identification of algae.

43

(i.e. binding strength). Algae are said to offer advantages for biosorption because of

their macroscopic particle structure, which provides a convenient basis for the

production of sorbent particles (Vieira and Volesky, 2000). Algae have a complex

cell wall as compared to bacteria and fungi, to the extent that within the algae group

we need to differentiate four evolutionary pathways, blue green algae, green algae,

brown algae, and red algae (Romera et al., 2006). There are studies reporting the use

of algae for the bioremediation of heavy metals, however there are only few reports

on the possible use of algae for the sorption of uranium (Vieira and Volesky, 2000).

Keeping these points in view algal biomass was procured either by culturing or

collected from natural habitats to test their uranium sorption potential.

3.2. Materials and Methods

3.2.1. Culturing of algae

Biomass for Chlorella vulgaris Beyerinck was raised in laboratory by scaling up the

culture in laboratory on Bold’s Basal Medium (Bold and Wyne, 1985) supplemented

with 10% soil extract. A continuous culture system for C. vulgaris was developed to

get a sufficient amount of biomass.

3.2.2. Collection and Identification of algae

The Maharashtra coast lies along the central west coast of India, between the

longitudes 15° 43’ to 20° N and latitude 72° 49’ to 77° 41’ E. The coast line is about

760 km long indented by estuaries, creeks, small bays, headlands, promontories, cliffs

etc. In the present survey, collection of algal biomass was done along the Maharashtra

sea coast from Colaba (Mumbai) in the north to Reddi in the south. The substratum

for the growth of algae on these coasts varied from lateritic to basaltic. In addition to

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the collections from sea coast collections from estuarine and fresh water bodies was

also carried out.

The material was identified by following the standard monographs and keys

(Desikachary, 1958; Desikachary et al., 1998; Krishnamurthy 2002, Mishra 1966).

Herbarium specimen of the material were prepared and deposited in the Herbarium,

Department of Botany, University of Pune, Pune, India. Part of the material was

preserved in 4% formalin for microscopic observations.

3.3. Results and Discussions

3.3.1. Culturing of algae

C. vulgaris (Chlorophyta) was grown on various synthetic media in order to check

which of the media was able to support the best growth of alga. Among the different

media growth of C. vulgaris was tested on BBM, CFTRI, and CHU 10. BBM was

observed to support the best growth of alga. When BBM was supplemented with 10%

soil extract, we could observe better growth. Hence for the subsequent experiments,

BBM supplemented with 10% soil extract was used for the growth of C. vulgaris.

Scaling up of the culture volume up to 10 liters was also done under controlled

photosynthetic conditions. Fig. 3.1 describes the continuous culture system that was

developed for the mass cultivation of C. vulgaris. Biomass was harvested by

centrifuging the culture at 6000 rpm for 10 minutes.

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Fig. 3.1. Continuous culture system

1.

2.

3.

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Fig 3.1. Continuous culture system for C. vulgaris

1. Medium reservoir.

2. Growth chamber.

3. CO2 supply

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3.3.2. Collection and Identification of algae

Altogether, various forms (microscopic, submicroscopic and macroscopic) of algae

growing in different habitats (marine water, estuarine water and fresh water), and on

different forms of substratum (lateritic, basaltic) were collected. 63 macroalgal

species belonging to 52 genera were collected. Among 63 different algae collected,

24 were from division Chlorophyta, 11 from Phaeophyta, 25 from Rhodophyta, and 3

from Cyanophyta. Among the 63 macro algae collected 49 species were collected

from marine water habitats, 12 species from fresh water habitats and 2 species from

brackish water habitats. The list of identified algae along with their habitat is given in

Table 3.1.

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7 2 ° 92°

8°

36°

Map of India and Study Area

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Table 3.1. List for algae collected from various habitats No. Name of the algae Habitat

Division Chlorophyta

1. Bryopsis pennata Lamouroux M.W.

2. Caulerpa peltata Lamouroux M.W.

3. Caulerpa sertularioides Gemlin M.W.

4. Caulerpa taxifolia (Vahl) C. Ag. M.W.

5. Chara wallichi Braun. F.W.

6. Chlorella vulgaris. Beyerinck. F.W.

7. Cladophora colabensis Boergesen B.W.

8. Cladophora attenuate (Bonnem.) J. Agardh M.W.

9. Cladophoropsis zollingeri (Kuetz) Boerg. M.W.

10. Enteromorpha prolifera (O.F. Mueller) J. Agardh M.W.

11. Enteromoprpha kylinii. Bliding. M.W.

12. Hydrodictyon reticulatum (L.) Lagerh F.W.

13. Monostroma oxyspermum (Kuetzing) M. Doty M.W.

14. Nitella hyaline (DC) Ag F.W.

15. Pithophora polymorpha Wittrock F.W.

16. Pseudobryopsis mucronata Borgesen M.W.

17. Rhizoclonium kochianum Kuetzing M.W.

18. Spirogyra crassa Kuetzing F.W.

19. Spongomorpha indica Thivy & Visalakshmi

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20. Tetrasporidium javanicum Mobius F.W.

21. Ulva fasciata Delile M.W.

22. Ulva lactuca Linnaeus M.W.

23. Valoniopsis pachynema (Martens) Boergesen M.W.

24. Zygnema cylindrosporum Czurda F.W.

Division Phaeophyta

25. Dictyota bartayresiana Lamouroux M.W.

26. Dictyopteris woodwardii (Brown) J. Agardh M.W.

27. Padina tetrastromatica. Hauck. M.W.

28. Sargassum crassifolium J. Agardh M.W.

29. Sargassum swartzii C. Agardh M.W.

30. Sargassum tenerrimum. J. Agardh. M.W.

31. Sargassum wightii (Greville) J. Agardh M.W.

32. Sphacelaria furcigera Kϋtzing M.W.

33. Spatoglossum asperum. J. Agardh. M.W.

34. Stoechospermum marginatum Kϋtzing M.W.

35. Giffordia mitchellae (Harvey) Hamel M.W.

Division Rhodophyta

36. Acanthophora delilei (C. Agardh) Greville M.W.

37. Amphiroa anceps (Lamarck) Decaisne M.W.

38. Bostrychia radicans Montagne M.W.

39. Caloglossa leprieurii (Montagne) J. Agardh M.W.

40. Compsopogon coeruleus (Balbis C. Agardh) F.W.

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41. Catenella repens Boergesen. E.W.

42. Centroceras clavulatum (C. Agardh) Montagne M.W.

43. Ceramium rubrum (Hudson) C. Agardh M.W.

44. Dasya iyengarii Boergesen M.W.

45. Gelidium micropterum Kuetz M.W.

46. Gelidium pusillum (Stackhouse) Le Jolis M.W.

47. Gracilaria corticata J. Agardh M.W.

48. Gracilaria foliifera (Forsskal) Boergesen M.W.

49. Grateloupia lithophila Boergesen M.W.

50. Hypnea musciformis (Wulf.) Lamouroux M.W.

51. Hypnea valentiae (Turner) Montagne M.W.

52. Jania rubens (Linnaeus) Lamouroux M.W.

53. Laurencia pedicularioides Boergesen M.W.

54. Porphyra vietnamensis Tanaka et Ho M.W.

55. Rhodymenia palmetto (Stackhouse) Greville M.W.

56. Sarconema filiforme (Sonder) Kylin M.W.

57. Scinaia furcellata (Turner) J. Agardh M.W.

58. Sirodotia cirrhosa Skuja ex. Balakrishnan et Chaugule F.W.

59. Solieria robusta (Grev.) Kylin M.W.

60. Spyridia filamentosa (Wulfen) Harvey M.W.

Division Cyanophyta

61. Scytonema bohneri Schmidle F.W.

62. Lyngbya majascula Harvey Gomout M.W.

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63. Nostoc linckia (Roth) Born Flah. F.W.

3.4. Conclusion

Biomass of 63 different algae could be procured either from collection or by

culturing. Along the northern part of Maharashtra, Mumbai in particular, industrial

development has threatened biological diversity. Nevertheless, there were certain

pockets that showed high macroalgal diversity and luxuriant growth. Perhaps,

environmental conditions prevailing at these pockets are conducive to their growth or

else, macroalgae have developed ability to withstand change in the environmental

conditions. Species belonging to genus Enteromorpha, (E. kylini) was reported, which

has not been reported from India so far.

In this study biomass was collected only one time from one site. However, a long

term monitoring and collection programme at selected sites may provide information

on how biodiversity changes are brought by natural means or by anthropogenic

impacts, which in turn may prove helpful for procuring large number of macroalgal

species.