THE CIJLTIVATIOX OF ALGAE &.q .z t+l. s-7Xf orw. z. . Az. # ..v. jxuo4'*'v.f: Xvs' c a. t '.t .y# âsu 4el y . . ..v.o ' g#*zh%,w. ..&x J f ' X 14 W >* * fev o'%'#' X.J : 1y:* fb.a . g # xy .4 .m, l+f w' y %vé +1 / +' ya *p, .. q 1 ... .%a/.G.. ,-* . # y zw,sx <' r''/?s 4. 4.r t .> *4 ' o yzz A -'.*z .*'2K t l3l 01 I (s Illlt g 11 elln THE CULTIYATION OF ALGAE ' By G. S. VEG TAMMAN K$z lckn INDIAN COUNCIL OF AGRICULTURAL RESEARCH wRsz ealwTlo DxczMsEa 1969 I ik Dtr6ctnr (1h41. W 16j .. C. G. RAOITAVA KVRIJP ; G'ltkfEditc : P. L. JM-AL , Auùtant Fzt#&r.' M. AmF ,,- N Chtkf Jhef:, 0.$c6,r.' KZr-AN KCMAR $ #p x z. .s. ,, oq6.o.à t * o po j : Noo ! Z #* l 9.# . oeï'l ql. ., . *z, Wz ts?pe/ jqrja 4... '- * @ 1969, 1.z 1* 1nd+ 6:relôV #' âgriœttvral JNz-Tâ, .#à?w Ibf/# Prmted m Indta by M C. Klmnnalz at tlïe Job Preo Pnmte Ltd , Kanplm aad publlshed by T S 'Prutlu, rnder-secretarp for the Indlatt Connral of Agnculturaj Rem-g.c,k New mlhi. 1. C. A. tt. MONQGRAPHS ON ALGAE i I EDITORTAL BOARD II R j M. S. ïta nt !lt a& va , 1) .E k: .,1? )ç.1., (:hiLl zr#,/,, ' 1 B P. Pal, Ph D., F.L.S., F.N.I., F.R.H.S., Mnnbet lj I T. V. Destkachary, PIZ.D., F,A.Sc,, F.N.l., Membc ' G. S. Venkataraman, Ph D., F.N.A.SC , Metttber FOREWORD This volume on tç The Cultlvatlon of Algae '> zs the nznth zn the series of monographs on algae mttlated by the lndlan Councll of Agrlcultural Research The extenslve rescarches on tlte varlous fundamentaland apphed aspects ofphycology carncd out dunng the past few decades have e-stabltshed the poterttlallty of algae as a sourcc of food, feed, fodder and manure as well as thelr role lll publlc health problems of a fast-developmg ctvlllzatlon The economlc utlhzatlon of algae has necessltated, ln recent years, the development of technlques to cultlvate them on a large sc-ale Except ln a few fortunate countrles, the number of the hungry, partlcularly ln the under-developed countrles of Asla, ls growlng faster than the supply of food Recent studtes have shown that some unicellular green algae lzke Chlorella cxan serve as a xotlrce of pz-oteln for huma.n and anlma,l consumptlon These organlsms, unllke the conventzonal food crops, can be culttvated all the year round, use the msxlmum solar eneqy and contam more protem and fats and less of mdlgestlble matter The fundamental process of gynthesls xn algae zs slmllar to that m any other chlorophyll-bcamng organlgm and a slngle Chkrella cell does exactly the samc ay a wheat or rlce plant, m terms of ultimate synthesls Technlques to cultlvate them on a large mdustrlal scale have been successfully developed ln some counhles and an attempt m thls dlrectlon ls worthwhlle lrr some of the undcr-developed countrles. ln recent years, blue-green algae have recelved conslderable attentlon ln mxny laboratorles, lncludlng the lhdian Agrlcultural Research Instltute, New Dellu, because of thelr tmportaacc In agnculturr Ccrtaln specles of algae carry out ly0th photosyntheszs and nltrogen ûxatton Large-scale culttvatlon of these algae and thelr appllcatlon to dte paddy fkelds have resulted ln lncrcased rlce pro- ductlon. Dr G S Vcnkauraman has beenworklng on tlus aspect for over a decade attd tlte results obtalned bg hIm are encouraglng Partzcularly m Indla, where rlce fornu a malor staple food for a large populatlon, the exploltatlon of these a4:1 ae in rlce cultlvatlon offers promlse The tzse of seaweeds as food, feed and manure ls another aspect tlzat requtres attentlon Wzth a coastllne as vast as ours, and an abundant supply of thcse algae, zt wlll be proûtable to lntroduce systematlc cultlvatlon ln the coastal regtons Thls wlll also result ln the development of a number of cottage lndustrlts ln the coastal areal. Indla has almost one-fourth of the total cattle populatlon of the world and tltere is a heavy competltlon between cattle and human belngs for food Eflbrts should, theretbre, be dlrected to find out hltherto unexplorcd resources of fodder and recourse may be taken to algae, at least ln coastal areas Thcse soweeds can also serve as an excelleat sourct of rnanure for a varlety of crops ln the coastal an*A.V. x FOREWORD Art lnteresttng recent developmznt in the cultwation of algae is sewage reclanutlon by the plwtosynthetm nucroalgae 'rhe sewage dzsposai problem m lndla ts tremendous and a large amount of tlus poterttza! source of manttre is wasted By growmg algae m sewage, tlie sewage Nvater can be reclalmed and the reclaimed water can be used for irrzgatmg crops The alral sltlrz'y can set-ve as nxanure and as a cheap source of protmn for poultrg and plgs. Bestdcs the large-scale cultwatlon of algae for econoznic utlllzationz laboratory culturas serv'e as tools for taxonoxnic, physlologlcal, blochemlcal and genetlc studles. I)r. Venuuraman is well known foc his work on the nitrogen 6wtlon by blue- green algae mt.lz speczal reference to thezr utlllzatzon ln agrzculture He ha6 dealt wlt,h vamous mspects of the cultlmtjon of algae, 130th for fundamental and applzed research, m thls volume. He ls to be congatulated on the mdustry and care wltlt whlc,h he has prepared this monograph, the need for wluch has been keenly felt for some tzme More than 2,500 references are llsted The author has also glven zmportant cltatlons under heads suclt as cytology, mutauon and genehcs, so that Interested workers ma,y congult them wlth ease. 1 hope students, teachers and research workers who are interested m tlze cultlvation of algae wlll find thks monopaph useful M S RANDHAWA i Vlce-chancellor Chandlgarh Punja'b Apicultural University 1 1 PREP' ACE This volume has been prepared as one of the serles of < Monographs on Algae ', - belng publlshed by the Indlan Counctl ot- Agrlcultural Researeh, New Delhz The sclzeme envlsages compzlatlon of a1l mformatlon pulllzshed so far on vaz lous aspects of Phycology Thls monograph has been prepared to partly meet ,uc.h a requlremgnt and also to prowde, especlally to the students of algae zn lndla, a workmg-gmde on the vamous aspects of algal ctllturmg The culttlrmg of algae as a research tool ls expandmg rapldly as modern rese.arc,h becomes mterested ln such dlverse ûelds as taxonomy, molphologv, pwhaytselro lsougpyp, lblelos,c hmeemdllsctlrnye,, tselsasuBee ecdu luttultllez, aftbloodn apnroddpuecrthloapn,s pagorwlecurlptruored,u wctalostne dlTsphoesualse, of controlled photosynthesls for nlalntalnlng the envtronment of space-shlp sultable labnrd h aftbmptcaatttolonn alls ctmome polfel tahtey l, ahtaevste d beevccolompem tehnet sv ehAlcllgega efo, rb )trm thpoelrrt astnrtu ecxtupreartl msymetkptsl zcalttkdy dscoverles In general, the methods of culturlng algae are the same as employed by the bacterlologlsts and the varlous modlficatlons ln the technlques are essentlally due to the speczal nature ol- algal cells It ls, therefore, zmportant to have a falr deglee of famlhamty aad proficleacy ln the bactenological technlqucs for a successful appllcatlon of these mcthoda foz' the cultumng of algae In the ûeld of morphology and taxonomyalgal cultures oflkr a definlte advantage ln preservlng the dellcate structures o,f the organlsms and zn reveallng tke extent of mrlablllty Our knowledge of the llfe hlstomes of algae has consl- derably expanded through the use of pule or um-algal cultures subterCraunltelzarne flmoreta hoodf s alhgaave,e sroemveea loef d waln4l chm taerree sotmf ga,g lblucut ltuhrltahle rlmtop ourntasunscpee cteAdn, example of the applzcatlon of pure culture technlques ls tlae dlscovery of thenltrogen- flxlng capaclty of certmn blue-green algaewhlch can be proûàbly explolted ln our general agrlcultural practlces as shown by, the author and other workers The past few ytars have wztnessed the use of algae m cultures for the aqsay of blologlcal and physlologtcal studzes relatlng to cancel lesearcll Llchen Vxonomy ls lncreaslngly supported by the cultura: studtes of the algal component of the symblosls Growth-promottag and growth-lnhlbltlng substances ploduced by some algae, wlazch havc a beanng cm tlze soll and aquatlc bzology as we:-l as on the % blologlcal equlllbrlum ' are all problems lnvolvlng cultural technlques Pule cultules of algae have proved convenlent mcdla for testlng the cilkcts oi varlous chemtcal agents on Ilvlng organlsms Thls has lts stgnlficancc I1l watel supplzcs and sanitatlon The tnductlon of synchrony ln a populatlon Ss a rnalor development ln the culture tcchnlques This has enabled us to undelstand vamous blologlcal processes xii plts>'wcs at a cellular level. Equally sigtufïcant ts the devclepmcnt of contlnuolzs fbw devlces for phpzologlcal research. Chaptm 1 and 9 glve detalls of these develoTphmee nfetsa.slbthty ofe an mvesttgation prlmxrily depends on the chotce, autlten- ttctty ard avallabtlity of the orghnlsm clzosen. Tiûs reqmres mamtenance of authentzc cultures m a genetlcally stable and vzable condltion. Although zn every laboratory, a stoc.k of algae is rnatntaaantd as a part of the research programme, only a few centres of collectlons have been establlshed, whlch mclude collectlons at Plymouth, Gottmgea, Prague, Cambmdge (England), Parzs, Bloomzngton (U S A ) and Tokyo At tlte Iftdlan Agrtcultural Research Instttute, New Dellzl, the author has recently se't up a culture collectton of mjcroalgaej the detaàls of whlch are gwcn ln AppEcvnedrl xs m1.ce Drs Wltsch and Spoeltr mdlcated the posslblllty of using fast- growmg unlcelluhr algae for food or feed, the algal cultures have assumed mdus- trlal dlmenslons Thc cultzvatzon of algac contlnuously under partlally or fully mmtrolled cortdtttolu ts knowa as T controlled phqtosynthesls ' and thls has become an lmpol tant development wtth great economic posnblhties A number of processes .. for cultlvatmg alTac ort a largc scale, both m morgamc soluuons and in waste waters, have been developed, the detxaib of which are glvea in Chapters 14 atzd 15 The lmmenslty of the scwage-dlsposal problem m Indla needs llttle cmphasls and blo,- loglcal reclalzlatltm of sewage deserve.s speclal consideration Equally important ls the large-scale productlon of cfliclent stralns of mtrogen-ftxlng blue-green algae for mcreaung the soll fertllzty, and also other useful algae whbch can be used econo- ' mlcally as a cattle or poultry feed. ln a hlghly organized and czvillzed socletyy where factors of productlon are dellcately balanced wjth our needs, a serious eflbrt to follow up any promlsing lead 1, certainly desîrable. Seaweed cultwauon for human food ln some countnes llke Japan has been deve- lopcd mto a full-fhdgqd agricultural enterpnse Wltll a coastline as vast as ourss these natural resources can be pro:tably explolted for fertlllzcr requirementg and ' for a vMarolreet yt hoaf uc o3t0ta gyee alrrls dhuastvrez ees.lapsed smce the publtcation of Kufreratlt's pape,,r h entTtled e La Culture des Alkucs ', which was subrequently followed m 1942 by l Boldt s exhaustlvc rewew on th- e < Cultlvatlon of Algae ,- azd ln l 946 by Prmgsheimv s valuable volume on f Pure Culture.s of Algae, tltelr Mamtenance and Preservatmn'. BesldeMhese, tizere has been a tremendous progress m this fteld of Pityqology, deallng wlt.h both fundamental and applled aspects Tlte present volume alms at presentlng the avallable lnformatlon on the metkods and applkcatzon of algal culture,s m a stmple, direct and conclse ferm, 5 havmg m mlnd tlle burdcns of the sttzdents and the worklng 1z11ca+1010g1st of today. Tlze cnormous expalulon oî research uslng algal cultures and the rapldlty wlth which new technlques and devlces are developed make lt fuhle to clalm any comprehen- slvcncss ln thls type of work Eflbrt has, however, been made to lnclude more than 2,500 cstxtmas and also to provtde lllustratlve matemals as far as possible. ln Chapter 8, specml methods for vamous classt.s of algae are gwen, based ort tlke avail- able lrtformatlon and on the autlwr's experience. PREFACK - xlll The :1st few chaptms deal mth the cultural techniques and requlrements Detxalled mformatton ls also lncluded on the nutmtlonal requirements of algae, nutrltlonal types and envzronmeatal factors, so as to glve a better comptehcnslon of the problems lnvolved In Appertdtx 11, cttattons of research papers based largely on algal tmltures are glven uader dlflkrent heads so that lnterested workers may refer to them To Tnclude every publlshcd work whlch ls m one way or the other based on algal culturcs zs obviougly lmposslble, for to covel from tlte penod of Famlntzln to the present day would demand not a book, but a lzbrary - Some common culture medla for algae and bacterla are gwen zn Appendzces III and IV respectlvely. 1 am zndebted to Dr M S Randhawa, Vce-chancellor, Punjab Agrlcultural University, Chandlgarh, for his znformed dlrectlon and helpful critlclsms durzng the preparatlon and publlcatLon of thl! volume I am also thankful to Dr B P Pal, Dlrector-General, Iadmn Councll of Agmcultural Research, Dr A B Joshz, Deputy Dlrector-General (Crops), Indlan Councll of Agrlcultural Research, Dr. M S Swanurmthan, Dlrector, Indlan Agrlcultural Research Institutey New Delhl, and Dr W V B Sundara lkao, Head of the Dlvlston of Mlcroblology, Indlan Agrlcultural Research Instltute, New Delhi, for thelr sustalned Interest and encouragement My smcere thanks are due to Prof R N Slngh of the Banaras Hlndu Unlverstty There ls much ln thls volume derlved from h1s teachlng, for wluch acknowledgement cannot be made by mcre clbatlon olx published work I am hlghly lndebied to Prof E G Prmgshelm of the Gottlngen Unwerslty, Germrmy, who generously placed hls entlre collectton of notes at my dlsposal They provlded me wlth a fund of lnformatlon, for whlch no amount of fotmal acknowltdgement will suëce. 1 am equally grateful to Prof. Atsushl Watanabe of the Smjo Unlverslty, Tokyo, Japan, whose valuable dzscusslons and znforma- tion on varlous aspects of algal culturlng were a consunt source of inspzlatton to me. Muc.h llclp has also been obtalned from the dlscussztms wlth Prof Tomomlchl Yanaglta, Dr Eljl Hase, Dr Y Fullta and Mr K Maruyama of the lnstltute of Applled Mlcrobloley, Unlverslty of Tokyo, Japan, dlzrmg my stay at thelr lnstltu- tmn I am also mdebted to Prof Y Yamada, Unlverslty of Hokkatdo, Sopporo, Prof Y. Naumura, lnstltute for Algologlcal Research, Muroran, Prof J Toklda, Faculty of Flsherlo, Unwerslty of Hokkaldo, Hakodate, and Prof A Arasakl, Unlverllty of Tokyo, for helpful dlscusslons and lnformatlon on the cultwatlon of marme algae m Japan To Drs Y Takechl and N Terashzma of the Japan Chlorella Company at Kunltachl, Tokyo, I am thankful for the informatlon on the nanqs cltlturlng of Chkrdla Mr S Uchlyama of the Research and Leglslatwe Department of the Natlonal Dlet Llbrary, TokyN, kmdly provlded the Informatlon on tlte totx'tl catch of seaweeds m Japan, for whlch I am gratrful My speclal thanks are due to Mr Otsuchl, Head of the Kakuni Flshermen Ccrporatloa and to Mr Kelshtro Kuda of the To1 Flshermen Corporatzon ln Hoklqaldo, Japan, for exphmmg to me the functlonzng and nunagement of seaweed farmtng Many sclentlst.s at home and abroad have contrlbuted exceilent photoglaphs and I am grateful to all of tllcm Al1 are thankfully acknowledged, as well as the