EVALUATION OF A MULTITROPHIC BIOFILTRATION SYSTEM WITH NEW
ALGAE SPECIES AND THE SEA CUCUMBER Holo hu ia sanc o i.
Luis, Felaco Du an
TESIS PRESENTADA Y DEFENDIDA PUBLICAMENTE PARA LA OBTENCIÓN DEL TÍTULO DE MASTER
OFICIAL EN CULTIVOS MARINOS
Las Palmas de G an Cana ia a 23 de Junio de 2014
III MASTER OFICIAL EN CULTIVOS MARINOS
Las Palmas de G an Cana ia, España
2012-2014
EVALUATION OF A MULTITROPHIC BIOFILTRATION SYSTEM WITH NEW ALGAE SPECIES
AND THE SEA CUCUMBER Holo hu ia sanc o i.
Luis, Felaco Du an
T abajo ealizado en las ins alaciones del Pa que Cien i ico Tecnologico de Talia e, el
ICCM y el Cen o de Bio ecnologia ma ina de Las Palmas de G an Cana ia, España, bajo la
di ección del D . Rica do Ha oun Tabaue y el D . Juan Luis Gomez Pinche i.
P esen ado como equisi o pa cial pa a la ob ención del Tí ulo o icial de Más e
Uni e si a io en Cul i os Ma inos o o gado po la Uni e sidad de Las Palmas de G an
Cana ia y del Diploma de Mas e o Science en Acuicul u a o o gado po el Cen o
In e nacional de Al os Es udios Ag onómicos Medi e áneos (CIHEAM).
Di ec o Di ec o Au o
Rica do Ha oun Tab aue Juan Luis Gómez Pinche i Luis Felaco Du an
III MASTER OFICIAL EN CULTIVOS MARINOS
O ganizado conjun amen e po la Uni e sidad de Las Palmas de G an Cana ia (ULPGC), el Ins i u o
Cana io de Ciencias Ma inas (Gobie no de Cana ias) y el Cen o In e nacional de Al os Es udios
Ag onómicos Medi e áneos (CIHEAM), a a és del Ins i u o Ag onómico Medi e áneo de Za agoza
(IAMZ)
Hay sol bueno y ma de espuma…
A mi abuela.
ACKNOWLEDGMENTS
A Dios.
A Venezuela que me ha hecho quien soy.
A mi amilia po se siemp e la más g ande mo i ación.
A Ana, po segui me sopo ando.
A BIOGES y su gen e, especialmen e a Ja ie po las e ulias sob e cambia el mundo.
Al GIA y oda su plan illa de in es igado es y écnicos po su disponibilidad y
p o esionalismo.
Al CBM-BEA y sus in es igado es y écnicos po la disposición de sus ins alaciones y
equipos.
A mis compañe os de mas e , po que más que unos colegas somos una amilia.
A Fe nando Tuya po el apoyo es adís ico y los consejos.
A Emilio Sole y Susy po el apoyo en axonomía.
A Ge cende po los consejos y po es a siemp e disponible.
A Dani y Paula po su amis ad y los buenos momen os.
A Rica do po la u o ía de es e abajo y su ce canía y amis ad.
i
CONTENTS
Page
ABREVIATIONS LIST
V
TABLES INDEX
Vi
FIGURES INDEX
Vii
ANNEXES INDEX
Viii
ABSTRACT
IX
1. INTRODUCTION
1
2. OBJECTIVES
12
3. METHODS
13
3.1. S udy a ea
13
3.2. Ini ial s a e o he bio il a ion acili ies
15
3.3. Assessmen o associa ed o ganisms
15
3.4. Species selec ion o cul u e ials
16
3.5. Cul u e ca e and ank design
16
3.6. Ammonia bio il a ion capaci y
19
3.7.In eg a ed cul u e o Holo hu ia sanc o i and Hyd opun ia co nea
20
3.7.1. Physiological s a us o he algae
22
3.7.2. Sea cucumbe sedimen bio il a ion capaci y
22
3.8. Holo hu ia sanc o i g ow h
23
3.9. Algae cul u e in opes
23
3.10. S a is ical analysis
24
4. RESULTS
25
4.1Ini ial s a e o he bio il a ion acili ies and associa ed o ganisms
25
4.2. Algae g ow h a e and p oduc ion
29
4.3. Algae bio il a ion
30
4.3.1. Bio il a ion ela i e o biomass
33
4.4. In eg a ed cul u e o Holo hu ia sanc o i and Hyd opun ia co nea
34
4.4.1. G ow h and p oduc ion a e
34
4.4.2. Bio il a ion, ammonia and physiological s a e
35
4.4.3. Sedimen il a ion by Holo hu ia sanc o i
38
4.4.4 Sea cucumbe g ow h and su i al
40
4.5. Rope cul u e expe imen
41
5.DISCUSSION
43
5.1. Ini ial s a o he bio il a ion acili ies
43
5.2. Algae g ow h and bio il a ion
44
5.3. In eg a ed cul u e o Holo hu ia sanc o i and Hyd opun ia co nea
46
5.4. Algae ope cul u e a he bio il a ion acili y
50
6. CONCLUSSIONS
51
7. RECOMMENDATIONS
52
8. REFERENCES
53
9. ANNEXES
61

ABREVIATIONS LIST
BCM
Bo anica Ciencias del Ma
FAO
Food and Ag icul u e O ganiza ion
GIA
G upo de In es igacion en Acuicul u a
ICCM
Ins i u o Cana io de Ciencias Ma inas
IMTA
In eg a ed Mul i-T ophic Aquacul u e
NUE
Ni ogen Up ake E iciency
NUR
Ni ogen Up ake Ra e
OM
O ganic Ma e
P
P oduc ion a e in d y weigh
PCT
Pa que Cien i ico Tecnológico de Talia e
SA
Su ace A ea
SD
S anda de ia ion
TAN
To al Ammonia Ni ogen
TOM
To al O ganic Ma e
ULPGC
Uni e sidad de Las Palmas de G an Cana ia
UV
Ul a iole
µ
A e age daily g ow h in pe cen age
i
TABLES INDEX
Page
Table 1. Table showing he ammonia concen a ions in µM ound a he acili ies o he
bio il e o he PCT.
26
Table 2. Mac oalgae species associa ed o he e luen s om bo h o he
aquacul u e ins alla ions loca ed in Talia e.
26
Table 3. Fauna associa ed o he e luen s om bo h o he aquacul u e
ins alla ions loca ed in Talia e.
27
Table 4. Weekly esul s summa izing he a e age amoun o ammonia a he en ance
o
he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e o
Ul a igida du ing he expe imen a ion pe iod wi h a densi y o 2g/L and a eno a ion
a e o 10 Vol/day.
31
Table 5. Weekly esul s summa izing he a e age amoun o ammonia a he en ance
o he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e
o
Colpomenia sinuosa du ing he expe imen a ion pe iod wi h an a e age densi y o
5.76g/L and a eno a ion a e o 10 Vol/day.
31
Table 6. Weekly esul s summa izing he a e age amoun o ammonia a he en ance
o
he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e o
Schizimenia dubyi du ing he expe imen a ion pe iod wi h an a e age densi y o
2,64g/L
and a eno a ion a e o 10 Vol/day.
32
Table 7. Weekly esul s summa izing he a e age amoun o ammonia a he en ance
o
he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e o Valonia
u icula is du ing he expe imen a ion pe iod wi h an a e age densi y o 4.47g/L
and a eno a ion a e o 10 Vol/day.
32
Table 8. S a is ical analysis o he a e age o each pa ame e o each species.
33
Table 9. Weekly esul s summa izing he a e age amoun o ammonia a he en ance
o he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e s
o Hyd opun ia co nea cul u ed wi h he e luen s om Holo hu ia sanc o i anks
du ing he expe imen a ion pe iod wi h a densi y o 4g/L and a eno a ion a e o 10
Vol/day.
36
Table 10. Weekly esul s summa izing he a e age amoun o ammonia a he en ance
o he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a es
o Hyd opun ia co nea cul u ed wi hou he e luen s om Holo hu ia sanc o i anks
du ing he expe imen a ion pe iod wi h a densi y o 4g/L and a eno a ion a e o 10
Vol/day.
36
Table 11. S a is ical analysis o each pa ame e e alua ed o H. co nea wi h o
wi hou he e luen s om he sea cucumbe anks.
37
Table 12. Su i al o Holo hu ia sanc o i in he expe imen a ion anks and he
con ol ank.
40
ii
FIGURES INDEX
Page
Figu e 1. Concep ual model o he ideal IMTA sys em ( om Fishe ies and Oceans
Canada 2013).
3
Figu e 2. Facili ies o he s udy A) G eenhouse whe e he cul u e anks we e loca ed
B)
Sedimen a ion ank o he acili ies o he complimen a y building 1 o he PCT C)
Bio il e acili y o he PCT.
14
Figu e 3. Species o algae es ed unde ank condi ions: A) Ul a igida, B) Codium
omen osum, C) Valonia u icula is, D) Colpomenia sinuosa, E) Dic yo a
mens ualis, F) Jania ubens, G and H) Schizymenia dubyi and I) Hyd opun ia co nea.
16
Figu e 4. De ails o he anks and some o he cul u es es ed in hem. A) Ul a igida, B)
Colpomenia sinuosa, C) cocul u e o Dyc io a mens ualis, Codium omen osum and
Valonia u icula is, D) cul u e o Codium omen osum, E) Schyzimenia dubyi, F) gene al
aspec o he anks.
18
Figu e 5. Holo hu ia sanc o i.
20
Figu e 6. De ails o he sea cucumbe se up A) he h ee anks co e ed in da k plas ic
bags o s imula e eeding beha io and a oid g ow h o unwan ed algae B) de ail
o he sedimen adding p ocedu e whe e he wa e le el is lowe ed C) de ail o
he pu e sedimen added as eed D) ela i e measu es o he sea cucumbe s in one
ank no ice he di e en sizes F) obse ed ep oduc i e beha io .
21
Figu e 7. Expe imen al se ing o e alua e he in e ac ion be ween sea cucumbe
anks and algae.
22
Figu e 8. Algae species and se up used o he ope cul u e expe imen A)
G a eloupia u u u u, B) G. imb ica a, C) Hyd opun ia co nea, D) ame wi h opes
and en wined algae, he i s ope ha e G. u u u u, he second G. imb ica a and he
hi d H. co nea.
24
Figu e 9. Wa e empe a u es du ing he s udy
25
Figu e 10. Fauna associa ed wi h he e luen s a he PCT and complimen a y building 1
o he PCT sedimen a ion ank. A) Osilinus a a us, B) Siphona ia pec ina a, C)
Ce i hium ulga um, D) Pa ella aspe a, E) Aplysia dac ilomela, F) Aip asia mu abilis, G)
Tunica es, H) Myxicola in undibulum, I) Eupolymnia nebulosa, J) example o an
agg ega ion o a ious o ganisms K) Example o escaped ish in his case, Se iola
dume ilii.
28
Figu e 11. G ow h a e p esen ed by he 4 species o algae e alua ed du ing h ee
weeks, s anda d de ia ion and a e age a e p esen ed
29
Figu e 12. P oduc ion a e p esen ed by he 4 species o algae e alua ed du ing
h ee weeks, s anda d de ia ion and a e age a e p esen ed.
30
Figu e 13. Bio il a ion in ela ion o biomass compa ing he ou algae species.
33
Figu e 14. Speci ic g ow h a e o Hyd opun ia co nea submi ed o he wo
ea men s showing he s anda d de ia ion.
34
Figu e 15. P oduc ion a e o Hyd opun ia co nea submi ed o he wo ea men s
35
Figu e 16. Physiological s a e in F /Fm o Hyd opun ia co nea submi ed o bo h
ea men s (wi h wa e om he e luen s o sea cucumbe o di ec ly om he
sedimen a ion ank ) du ing he pe iod o s udy.
37
Figu e 17. A e age amoun o ammonia a he en ance and exi o each week
measu ed a he en ance and exi o he sea cucumbe anks and i s di e ence.
38
Figu e 18. Amoun o o ganic ma e (g/100g) p esen a he beginning and he
end o each week du ing he s udy.
39
Figu e 19. To al ni ogen (mgN/100g) p esen in he sedimen s a he beginning and
39
iii
he
end o each week o ea men .
Figu e 20. Shows he di e ence in he weigh o Holo hu ia sanc o i o each ank
a e 100 days o eeding wi h sedimen om he sedimen a ion ank.
40
Figu e 21. A e age weigh o Hyd opun ia co nea, G a eloupia u u u u and G.
imb ica a a he beginning o he ope expe imen and a e 40 days.
41
Figu e 22. A e age g ow h a e o Hyd opun ia co nea, G a eloupia u u u u and
G. imb ica a a he beginning o he ope expe imen and a e 40 days.
41
Figu e 23. ini ial and inal s a e a e 40 days o he opes s ocked wi h G acilla ia
co nea, G a eloupia imb ica a and G a eloupia u u u u a he bio il e o he PCT.
42
ANNEXES INDEX
Page
Annex 1. Example o he iden i ica ion p ocess o Schizymenia dubyi using i s
mac oscopic and mic oscopic mo phology. A) mo phology o he hallus wi h he
e y small s ipe B) Mo phology o hallus in na u al condi ions, C) C oss sec ion
o he blade showing epiphy es ( igh a ow) and medula ilamen s (le a ow), D)
C oss sec ion o he blade showing ea ly de elopmen o he gonimoblas (le a ow),
E) c oss sec ion showing he os iole in a well-de eloped gonimoblas and he expulsion
o spo es (le a ow), F) de ail o he spo es (le a ow).
61
Annex 2. P oposed mechanism o inc ease he su ace a ailable o algae g ow h a
he
bio il e o he PCT A) ini ial s a e, B) condi ion a e a mon h iewed om he su ace
and C) condi ion a e a mon h illed wi h Colpomenia sinuosa a he con inuously
subme ged side.
62
6
ca bohyd a es in b own seaweeds ac as soil condi ione s imp o ing ae a ion and soil
s uc u e, especially in clay soils, and ha e good mois u e e en ion p ope ies. Some
s udies also show ha ex ac s o b own algae can imp o e g ow h a es and educe
pes s, consequen ly inc easing c op yields as well as imp o ing he o e all quali y o he
p oduc (Blunden e al. 1996; Leach e al. 1999).
A mo e ecen app oach o seaweed use is as sou ce o uel and as ca bon sinks
(Packe . 2009), conside ing he possible use o la ge a eas o cul i a ed seaweed, as a ool
o mi iga e global wa ming (Mu aoka. 2004). CO2 acquisi ion by ma ine mac oalgae can
ep esen a conside able sink o an h opogenic CO2 emissions and ha es ing and
app op ia e use o mac oalgal p ima y p oduc ion could play a signi ican ole in ca bon
seques a ion and amelio a ion o g eenhouse gas emissions (Chung e al. 2011).
A p esen he e a e app oxima ely 200 species o seaweeds used wo ldwide
(Zemke - Whi e and Ohno 1999), o which he ollowing species o gene a a e in ensi ely
cul i a ed: Lamina ia japonica, Unda ia pinna i ida, Po phy a spp., Eucheuma spp.,
Kappaphicus al a ezii, Hyd opun ia spp., Monos oma spp. and En e omo pha spp.
(Lüning and Pang 2003).
The inabili y o adi ionally high alue gene a such as Hyd opun ia, Po phy a and
mo e ecen ly a ge ed high yield species such as Aspa agopsis (Dawes e al. 1999; Ma os
e al. 2006 ) o su i e he b oad en i onmen al luc ua ions o opical pond - based
sys ems p o ides an impe us o he selec ion o al e na i e bio emedia ion species
(Muñoz e al. 2011).
Mos o he echnology o in eg a ed mul i - ophic aquacul u e (IMTA)
s a egies al eady exis s, howe e i has no been p ope ly implemen ed in a la ge,
ma ke able, comme cial scale (Buschmann e al. 2009 ). Fu he mo e, seaweed
bio il a ion o ish a m e luen s has no been adop ed by he aquacul u e indus y.
Since mos seaweeds a e comme cially less aluable han cul u ed ma ine
animals, and conside ing ha one o he mos cos ly componen o die s used in
aquacul u e is p o ein (Neo i e al. 1998), One p oposed way o inc ease he economic
iabili y o seaweed bio il e s has been o eed he biomass p oduced o comme cially

7
aluable mac oalgi o es which can con e la ge quan i ies o he low- alued seaweed
in o highly alued commodi ies (Shpigel and Neo i, 1996) such as sea u chins and
abalone.
The nu i ional alue o seaweeds di e s acco ding o he species and he amily
conside ed. Fo example, he ed seaweeds con ain a high le el o p o eins (Fleu ence,
1999; Fujiwa a-A asaki e al. 1984) while he g een seaweeds, such as Ul a lac uca, show
a lowe p o ein le el and b own algae, such as L. japonica, ha e he lowes p o ein
con en . Howe e , seaweed ea ed in ishpond was ewa e e luen s egula ly inc ease
hei p o ein con en and p esen a signi ican ly highe die a y alue compa ed o algae
ea ed in egula sal wa e . This ansla es o be e g ow h pe o mance as expe imen s
in Halio is ube cula a coccinea (Vie a e al., 2011) and o he comme cially impo an
species such as he sea u chins Psammechinus milia is and Pa acen o us li idus (Cook
and Kelly, 2006) ha e showed.
These imp o emen s a e signi ican conside ing ha a limi ing ac o o u he
expansion o abalone and o he mac oalgi o es aquacul u e is he es ic ed a ailabili y
o an economically and en i onmen ally sus ainable eed, as hese cul u es equen ly
equi e la ge quan i ies o wild ha es ed mac oalgae (Vie a e al., 2005) and ha animals
ed mixed die s pe o m signi ican ly be e han hose ed a single algal die (Vie a e al.,
2011).
The use o mac oalgae as bio il e s and as aluable ma icul u e p oduc s is no
limi ed o land based ins alla ions, an al e na i e app oach in seaweed pond and di ch
cul u e, using s akes and opes, has eme ged om he echnologies used o la ge-scale
coas al seaweed a ming o Sou heas Asia. This app oach, howe e , has no p o ided he
nu ien emo al a es and he a eal yields necessa y o in ensi e ma icul u e, o
easons p obably ela ed o insu icien wa e u bulence, high u bidi y and low nu ien
concen a ions, howe e , wi h he inc ease o in e es in o sho e aquacul u e, hey ha e
been shown o wo k e en in ha sh o sho e condi ions (Buck and Buchholz. 2004, Neo i e
al. 2004, Ga cia 2012; Leonczek 2013) mos ly using long line seaweed aquacul u e
echniques inc easing i s po en ial as in eg a ed mul i ophic aquacul u e species.
8
De i i o es in IMTA
Issues such as solid was e managemen , nu ien ecycling and eed con e sion
enhancemen a e also mo e easily and p o i ably add essed on an indus ial scale on land
han in open-wa e ish a ms (Neo i e al., 2004), howe e , wi h he expansion o cage
aquacul u e, he e is a need o ea ing hese solid was e di ec ly in open wa e
condi ions, ha is he eason why o he complemen a y in eg a ed app oach o he
educ ion o he en i onmen al impac on he sea bo om by he ne pen sludge has been
he cul u e unde nea h hem o sca enge s like sea cucumbe s as seconda y c ops
(Ahlg en, 1998) using hem as so o animal bio il e o sedimen s.
In his way, he in eg a ed mul i ophic aquacul u e app oach is no limi ed only
o he p oduc ion o mac oalgae om he e luen s o adi ional eed aquacul u e, i has
also been es ed as ood sou ce o suspension eede s and de i i o es, his gains
impo ance conside ing ha he mos signi ican pollu ing componen s o aquacul u e
e luen s is no only ammonia, bu also, suspended solids (To a e al. 2000).
O ganic ouling deb is made up o ish eces, excess ish ood, algae, in e eb a es,
and o he pa icula e o ganic ma e can accumula e along he sides and loo o he
loa ing pens and on he sea loo below, clogging he ne s and es ic ing wa e
ci cula ion and possibly deple ing oxygen in he wa e and sedimen s which in u n can
s ess ish. The eeding habi s o some species o sea cucumbe s can alle ia e ouling
deb is p oblems a salmon ma icul u e acili ies his was demons a ed by Ahlg en in
1998 who conduc ed expe imen s in which ed sea cucumbe s Pa as ichopus cali o nicus
we e allowed o eed inside loa ing ne pens a a salmon ea ing acili y in Sou heas
Alaska. These expe imen s showed ha sea cucumbe s consumed ouling deb is and
clea ed a signi ican amoun o su ace a ea on he ne s.
By doing his, sea cucumbe s can ake ad an age o much o he ouling deb is
which is de i ed om p o ein- ich o ganic ma e such as ish ood, bo h be o e and a e
i has passed h ough a ish gu and e en dead ish y, u ning ha m ul deb is in o a
ma ke able p oduc (Ahlg en. 1998).
9
These impac s do no happen only in ish aquacul u e, i has been demons a ed
ha dense assemblages o il e - eeding bi al es enhance he e ical low o o ganic
ma e owa ds he ben hic en i onmen . Palza e al. (2008) ga he ed e idence ha sea
cucumbe s will u ilize, and he e o e educe, he ben hic o ganic deposi ion om oys e s
gi en ha when g own in co - cul u e showing bo h ac i e selec ion o o ganic ma e ial
om he sedimen s and diges ion/assimila ion o hese o ganics in he gu . This sugges s
he easibili y o de eloping a comme cial-scale co-cul u e sys em ha would bo h educe
he amoun o o ganic deposi ion unde nea h shell ish a ms and p oduce a seconda y
cash c op (Pal za e al. 2008).
The e ha e also been ials o he polycul u e o sh imp and sea cucumbe s
(Holo hu ia scab a) bu hei success has been limi ed. In se e al ials, i seemed ha
sh imp caused he dea hs o sea cucumbe s, and in some cases i was clea ha hey did
so (Pi e al. 2004). In o he ials, su i al and g ow h o sand ish ea ed wi h sh imp o
3 weeks we e signi ican ly lowe han o sand ish ea ed alone. Con i med ha co-
cul u e is no iable. All sand ish ea ed in co-cul u e we e dead o mo ibund a e a
mon h (Bell e al. 2007).
Some species o sea cucumbe s ha e been epo ed o be able o ea up o 76 % o
To al o ganic ma e (TOM). Zamo a and Je s in 2011 demons a ed he abili y o he sea
cucumbe Aus alos ichopus mollis o use di e en le els o TOM o gene a e simila
g ow h a es mainly by changes in hei eeding beha io and diges i e physiology. The
TOM, in he biodeposi s sampled benea h mussel a ms anged om 2.8% o 18.2% which
we e wi hin he ange co e ed in he labo a o y eeding expe imen s (Zamo a and Je s,
2012). O he die s ha e been es ed by Yuan and collabo a o s (2006), hey epo ed ha
a mixed die con aining mos ly mussel eces and 25 % powde ed algae showed he bes
s anda d g ow h a e.
In he Cana y Islands he main ep esen a i es o Holo hu ians a e Holo hu ia
mamma a, Holo hu ia a guinensis and Holo hu ia (Pla ype ona) sanc o i Delle Chiaje
1823 he la e species is he mos abundan in he a chipelago and e y conspicuous in
he Medi e anean and adjacen eas e n A lan ic whe e i has comme cial alue (Aydin,
2008). I shows a ma kedly noc u nal beha io , wi h la ge dis ances and as es
10
displacemen s du ing he end o he nigh ime, emaining hidden in c acks and c e ices
du ing dayligh and mo ing a nigh o eed (Pé ez-Ruza a e al. 1984; Pé ez-Ruza a and
Ma cos 1987) H. sanc o i is one o he mos selec i e species when i comes o he
selec ion o o ganic ma e om he sedimen s (Mezali and Soualili. 2013).I s
consump ion o OM has been shown o inc ease wi h OM a ailabili y, pa icula ly du ing
o ma ion o he gonads (Na a o e al. 2013a).
Economic alue and uses o sea cucumbe s
Sea cucumbe s, can be e y ma ke able speci ically on hei d ied o m also called
beche-de-me hey ha e been a die a y delicacy and medicine o Asians o e many
cen u ies. The collec ion o sea cucumbe s o supply he ma ke has seen a deple ion o
his esou ce in he adi ional ishing g ounds close o Asia and mo e ecen ly he
expansion o his ac i i y o new and mo e dis an ishing g ounds. Cu en ly, he e a e
ishe ies ha es ing sea cucumbe s ac oss mos o he esou ce ange, including emo e
pa s o he Paci ic, he Galapagos Islands, Chile and he Russian Fede a ion. Sea
cucumbe s ocks a e unde in ense ishing p essu e in many pa s o he wo ld and
equi e e ec i e conse a ion measu es. Sea cucumbe s p o ide an impo an
con ibu ion o economies and li elihoods o coas al communi ies, being he mos
economically impo an ishe y and non- in ish expo in many coun ies. A mul i ude o
sea cucumbe species a e being exploi ed wo ldwide, wi h new species being b ough o
ma ke as es ablished species become sca ce and mo e di icul o ind. Li le is known
abou he ecology, biology and popula ion s a us o mos comme cial species. (To al-
G anda e al., 2008)
The majo i y o sea cucumbe s a e expo ed o he beche-de-me ma ke and ew
species o he li e ade (aqua ium) ma ke . In many coun ies, pa icula ly in he
Wes e n Paci ic egion, some sea cucumbe s o hei o gans a e consumed as delicacies
o as a p o ein componen o adi ional die s (To al-G anda e al. 2008). They ha e high
comme cial alue coupled wi h inc easing global p oduc ion and ade. Sea cucumbe s
ha e long been used o ood and medicine in he communi ies o Asia and Middle Eas .
Nu i ionally, sea cucumbe s ha e an imp essi e p o ile o aluable nu ien s such as
Vi amin A, Vi amin B1 ( hiamine), Vi amin B2 ( ibo la in), Vi amin B3 (niacin), and
11
mine als, especially calcium, magnesium, i on and zinc. A numbe o unique biological and
pha macological ac i i ies including an i-angiogenic, an icance , an icoagulan , an i-
hype ension, an i-in lamma o y, an imic obial, an ioxidan , an i h ombo ic, an i umo
and wound healing ha e been desc ibed o a ious species (Bo dba e al. 2011)
Aquacul u e, sea anching and es ocking ha e been e alua ed as possible
solu ions o wild sea cucumbe o e exploi a ion, and some coun ies ha e s a ed such
en u es (e.g. Aus alia, China, Ki iba i, Philippines, Vie Nam and Madagasca ) China is
success ully p oducing an es ima e o 10,000 onnes d y weigh o Apos ichopus japonicus
om aquacul u e, mainly o supply local demand (To al-G anda e al., 2008).
In he las yea s, he esea ch g oup in Aquacul u e o he ULPGC has been
wo king on bio il e ing p ocesses o he e luen s om he aquacul u e p oduc ion uni in
he Complemen a y Module 1 o he Scien i ic and Technological Pa k. The main ocus
has been ew seaweed species as nu ien s ipe s and as sou ce o biomass o abalone
p oduc ion. To ou knowledge, he e has no been any s udy whe e he e luen s
collec ed a he sedimen a ion anks o in land aquacul u e acili ies a e used o eed
de i i o es, u he mo e, he in e ac ion be ween sedimen eede s, wa e column and
cul u ed algae has no been ho oughly s udied.
This s udy was s a ed o enhance he po en ial use o new mac oalgae o be
inco po a ed in he bio il e ing sys em as well as o de e mine he ole o sea cucumbe
as a sedimen o ganic ma e consume . The combina ion o hese wo ypes o nu ien
emo e o ganisms (bio il e o ganisms) has no been e alua ed ye and he need o
unde s and he possible syne gy be ween sea cucumbe and seaweed aquacul u e a e
he main ele an new ac o s in his wo k. In his sense, 3 main esea ch lines ha e been
opened a ound he concep o in eg a ed aquacul u e sys ems o allow en i onmen al
iendly and mo e p o i able land-based aquacul u e p oduc ion. The e is a g ea need o
di e si y cul i a ed seaweeds ha could be used as bio il e s wi h ma ke iabili y and i is
impo an o de e mine he e iciency o il a ion o hese seaweeds, hese a e key
ac o s ha a e add essed in his wo k.

12
2. OBJECTIVES
Gene al objec i e:
The gene al objec i e o his s udy was o e alua e and implemen po en ial new ma ine
species wi h sedimen and wa e bio il a ion capaci ies o he nu ien s om a land-
based aquacul u e p oduc ion uni .
Speci ic objec i es:
In o de o accomplish he main objec i e o his s udy se e al speci ic objec i es whe e
designed
 Iden i y he main o ganisms p esen in e luen s om he in land aquacul u e
acili ies in he Scien i ic and echnologic pa k o Talia e (Pa que Cien i ico
Tecnológico de Talia e, PCT) ocusing on he Bio il a ion po en ial o he
o ganisms.
 E alua e he ini ial s a e o he bio il a ion acili y in he Pa que Cien í ico
Tecnológico de Talia e PCT and make he possible and necessa y modi ica ions o
imp o e i .
 De e mine he bio il a ion capaci y o he mos abundan species o algae p esen
in he e luen s as well as hei p oduc ion and esis ance o ee loa ing cul u e
sys ems.
 Assess he iabili y o adding Holo hu ia sanc o i as a way o bio il a e he
sedimen s om he sedimen a ion ank a he PCT and inc ease he nu ien s
a ailable in he wa e o he g ow h o algae in ee loa ing sys ems.
 De e mine he g ow h and su i al o Holo hu ia sanc o i ed wi h sedimen s om
he sedimen a ion ank o he e luen s o he PCT.
 Tes he po en ial use o algal ope cul u e o cul i a e algae wi h comme cial
in e es in he bio il e ins alla ion o he PCT.
13
3. MATERIALS AND METHODS
3.1. Bio il a ion acili ies:
This s udy was made a he in eg a ed cul u e acili ies o he esea ch g oup in
aquacul u e (GIA; ULPGC) and he Pa que Cien i ico Tecnológico de Talia e (PCT) loca ed
in he eas e n side o he Island o G an Cana ia, in Talia e, Telde. The expe imen al
anks (desc ip ion o anks on 3.5) whe e loca ed inside a g eenhouse in he acili ies o
he complimen a y building 1 o he PCT (Figu e 2 A); he g eenhouse ecei ed wa e
h ough a pump loca ed a an 11 m3 sedimen a ion ank ha collec s he suspended
pa icles o all he e luen s om he cul u e o a ious species o he acili y (Figu e 2 B).
On he o he hand, a he PCT´s acili ies, he bio il e acili y loca ed ou doo s
consis s on an elonga ed ank sys em wi h a s ai case design ha c ea es cascades o
wa e on each le el, he app oxima ed o al olume o his bio il e is o 160 m3 wi h
app oxima ely 30 eno a ions o i s olume pe day. The e luen s om h ee ish
p oduc ion acili ies en e s ough 3 pipes loca ed in he uppe side o he bio il e a he
cen e , i has a small sedimen a ion ank o app oxima ely 1.60 m deep on one o he
sides o he i s le el, om his i s le el he wa e alls by g a i y o 8 o he le els ha
a e made o conc e e anks o app oxima ely 30 cm wide, 40 cm deep by 14 m long. The
bio il e ends in a cascade o app oxima ely 2.3 m a e which he wa e lows di ec ly o
he ocean (Figu e 2 C).
14
Figu e 2: Facili ies o he s udy A) G eenhouse whe e he cul u e anks we e loca ed B)
Sedimen a ion ank o he acili ies o he complimen a y building 1 o he PCT C) Bio il e
acili y o he PCT.
A
B
C
15
3.2. Ini ial s a e o he bio il a ion acili ies
The ini ial s a e o he bio il a ion acili y was de e mined ia di ec obse a ion
o bo h o he designa ed bio il e ins alla ions ( he complimen a y building 1 o he PCT
and he main building) specially aking in o accoun he sedimen a ion pa e ns and he
di ec ion o he wa e low.
In o de o imp o e he di ec ion o he wa e low and he sedimen a ion o
pa icles (which was oo as and in he cen e o he bio il e ) a se ies o pipes whe e
connec ed o he wo main exi s, edi ec ing hei low owa ds he sedimen a ion ank
loca ed on he igh side o he en ance o he wa e o he bio il e . The i s se o pipes
was connec ed o he i s en ance o he wa e o he sys em which was he one wi h
he s onges wa e low, his modi ica ion enabled he wa e o en e he bio il e nex
o he sedimen a ion ank, he second se o pipes was connec ed o he second en ance
o wa e loca ed app oxima ely 30 cm o he le o he i s en ance and 15 cm deepe ,
he e he wa e low was edi ec ed also owa ds he sedimen a ion ank, bu unde he
i s se o pipes.
Wa e samples we e collec ed in he le , cen e and igh a ea a he en ance
and he exi o he wa e h ough he sys em o de e mine o al ammonia o he
bio il a ion acili y a he PCT.
3.3. Assessmen o associa ed o ganisms
A apid census o he o ganisms associa ed wi h he acili y was made by eco ding
e e y mac oscopic o ganism seen along each le el o he bio il e a he PCT and he
sedimen a ion ank o he complimen a y building 1 o he PCT acili y, pic u es and
samples we e aken o each species wi h emphasis in algae species, which we e collec ed
and placed on he e e ence collec ion o he he ba ium “He ba io BCM (Bo anica
Ciencias del Ma )”.
Each species was iden i ied using specialized axonomic e e ences based on he
o iginal desc ip ions and on a c i ical analysis o he li e a u e and in he case o algae,
s e eo and compound mic oscopes we e used o look and desc ibe he mo phological and
ana omical cha ac e s o he p ope iden i ica ion o he species.
22
Figu e 7: Expe imen al se ing o e alua e he in e ac ion be ween sea cucumbe anks
and algae.
3.7.1. Physiological s a us o he algae:
The physiological s a us o Hyd opun ia co nea submi ed o bo h ea men s was e i ied
by measu emen o pho osyn he ic e iciency o chlo ophyll (F /Fm) by he luo escence
emission using he Hansa ech Pocke PEA Chlo ophyll luo ime e .
3.7.2. Sea cucumbe sedimen bio il a ion capaci y.
Two su ace sedimen samples o app oxima ely 100 ml we e aken om each
ank he day a e sedimen was pou ed in and a e a week by in oducing a u ine sample
bo le o 100ml a abou 1 cm in he sedimen a a andom place o he ank, and hen
mo ing i o abou 5 cm in a s aigh line, each sample was ozen un il all he samples o
he 3 days ial we e collec ed, hen hey we e d ied in a s o e a 100 ºC o
app oxima ely 24 hou s un il hey eached cons an weigh , hen each sample was
homogenized manually o abou 20 minu es, each analysis was made in iplica es, he
me hods used o he analysis a e as ollows.
Pe cen age o o ganic ma e was calcula ed by bu ning app oxima ely 4 g o
sedimen sample in a Mu le u nace a 600 ªC du ing a leas 4 hou s (AOAC, 2005).

23
To al ni ogen in he sedimen s was analyzed using he me hod desc ibed by
Kjeldahl (AOAC, 2005), which consis s on he diges ion o he samples a 420 ªC wi h
concen a ed sul u ic acid in p esence o a me cu ic ca alyze du ing an hou , a e which
he sample is dis illed wi h 40% NaOH wi h bo ic acid as a ecei e subs ance (using he P
Selec a Bloc Diges o he diges ion and he P Selec a Kjeldahl dis ille p o ni o M o he
dis illa ion), in he end, he sample is i a ed wi h HCl 0.1 N and he amoun o ni ogen
is calcula ed acco ding o he o mula
N (mg/100g)=ml o i a ion – a e age pa e n i a ion ml) x 0.1 (mola i y o HCl) x 14.007
mg o sample x 100
3.8. Holo hu ia sanc o i g ow h
T ee anks o sea cucumbe s we e ed e e y 15 days (excep o he pe iod o 3
weeks when he expe imen wi h H. co nea ook place, when hey we e ed weekly, also
keeping a con ol ank wi h no eed) hey we e weighed a he beginning and hen a he
end o he 100 days expe imen and moni o ed each week.
3.9. Algae cul u e in opes.
Th ee PVC ames o 35 cm by 1 m we e c ea ed, h ee 1 m opes we e placed in
each ame and each ope was s ocked wi h ei he G a eloupia imb ica a, G a eloupia
u u u u o Hyd opun ia co nea (Figu e 5, A, B and C), so ha each ame had one species
pe ope, he algae we e pu in he opes by en wining he hallus in o he ope (Figu e 8
D). Each ame was placed in he same le el o he bio il e ins alla ion a he PCT a he
le , cen e , and middle sec ion o ensu e all possible combina ions and o educe he
e ec o he loca ion inside he bio il e , hey we e ha es ed a e 40 days unde hose
condi ions.
24
Figu e 8: Algae species and se up used o he ope cul u e expe imen A) G a eloupia
u u u u, B) G. imb ica a, C) Hyd opun ia co nea, D) ame wi h opes and en wined
algae, he i s ope ha e G. u u u u, he second G. imb ica a and he hi d H. co nea.
3.10. S a is ical analysis.
Non pa ame ic s a is ics was used because he da a does no comply wi h he equisi es
o pa ame ic s a is ics, he U o Mann Whi ney Tes and analysis o a iance ia
pe mu a ions (PERMANOVA) whe e used o e alua e i he e we e signi ican di e ences
be ween ea men s and species.
A
C
B
D
25
4. RESULTS
4.1. Ini ial s a e o he e luen s and associa ed o ganisms.
The empe a u es du ing he s udy a e shown in igu e 9, hey a ied be ween 24
o 20 ºC. The To al ammonia associa ed wi h he bio il e a he main building o he PCTT
did no di e a he en ance and exi o he acili y ( able 1).
The o ganisms associa ed wi h he e luen s o bo h acili ies di e ged, in gene al,
he sedimen a ion ank o he complimen a y building 1 o he PCT had mo e species
ichness, 11 mac oalgae (Table 2) and 16 mac o auna species (Table 3) while he bio il e
a he PCT only p esen ed 7 species o mac oalgae (Table 2) and 14 species o mac o auna
(Table 3).
Rega ding mac oalgae g oups, he e was a dominance o species o ed algae in
he e luen s om he complimen a y building 1 o he PCT wi h 5 species, ollowed by
ep esen a i es o g een algae wi h 4 species, meanwhile, in he bio il e ins alla ion a
he PCT, B own algae domina ed oge he wi h g een algae, bo h wi h 3 species (Table 2).
The associa ed auna p esen ed he same ichness in bo h acili ies, wi h 16
species. The mos p e alen and dominan g oup we e he mollusks wi h 5 species a he
ins alla ion in he PCT and 7 a he sedimen a ion ank o he complimen a y building 1 o
he PCT acili y (Table 3), ( igu e 10).
Figu e 9: Wa e empe a u es du ing he s udy
15
17
19
21
23
25
27
10/7/2013 11/7/2013 12/7/2013 1/7/2014 2/7/2014 3/7/2014 4/7/2014
Tempe a u e (ºC)
Da e
26
Table 1: Table showing he ammonia concen a ions in µM ound a he acili ies o he
bio il e o he PCT.
En ance
Exi
Le
8.485
8.345
Middle
9.467
8.626
Righ
11.289
10.168
A e age
9.747
9.046
S de
1.161
0.801
Table 2: Mac oalgae species associa ed o he e luen s om bo h o he aquacul u e
ins alla ions loca ed in Talia e.
Species
Complimen a y building 1
Bio il e PCT
Phaeophy a
Dyc io a mens ualis
Dic yo a mens ualis
Colpomenia sinuosa
Colpomenia sinuosa
Pla oma Sp.
Rhodophy a
Jania ubens
Jania ubens
Schizymenia dubyi
G a eloupia imb ica a
G a eloupia u u u u
Hyd opun ia co nea
Chlo ophy a
Ul a igida
Ul a igida
Valonia u icula is
Valonia u icula is
Codium omen osum
Codium
omen osum
Caule pa acemosa
To al ichness
11
7
27
Table 3: Fauna associa ed o he e luen s om bo h o he aquacul u e ins alla ions
loca ed in Talia e.
Species
Complimen a y building 1
Bio il e PCT
Po i e a
Spongionella pulchella
Cnida ia
Aip asia mu abilis
Aip asia mu abilis
Cho da a
Ascidiacea Sp
Ascidiacea Sp
Echinode ma a
Holo hu ia sanc o i
Coscinas e ias enuispina
Coscinas e ias enuispina
Ophiode ma sp
Ophiode ma sp
Annelida
Polychae a sp.
Eupolymnia nebulosa
Sipunculus nudus
Sipunculus nudus
Myxicola in undibulum
Molusca
Aplysia dac ylomela
Aplysia dac ylomela
Pa ella aspe a
Pa ella aspe a
Ce i hium ulga um
Ce i hium ulga um
Osilinus a a us
Osilinus a a us
Siphona ia pec ina a
Siphona ia pec ina a
Halio is ube cula a coccinea
Aplysia depilans
A h opoda
Pachyg apsus
ma mo a us
Ve eb a a
Spa us au a a
Spa us au a a
Se iola i oliana
Dicen a chus lab ax
To al ichness
16
16

28
Figu e 10: Fauna associa ed wi h he e luen s a he PCT and complimen a y building 1 o
he PCT sedimen a ion ank. A) Osilinus a a us, B) Siphona ia pec ina a, C) Ce i hium
ulga um, D) Pa ella aspe a, E) Aplysia dac ilomela, F) Aip asia mu abilis, G) Tunica es, H)
Myxicola in undibulum, I) Eupolymnia nebulosa, J) example o an agg ega ion o a ious
o ganisms K) Example o escaped ish in his case, Se iola dume ilii.
C
A
E
D
F
G
B
I
H
J
K
29
4.2. Algae g ow h a e and p oduc ion.
The expe imen s we e ca ied ou be ween he 15/10/2013 and he 07/11/2013
o Colpomenia sinuosa and Ul a igida and o Schizymenia dubyi and Valonia u icula is
be ween he 27/01/2014 and he 18/02/2014.
E e y species o algae e alua ed showed a endency o dec ease du ing each
expe imen a ion pe iod, he species ha showed he highe speci ic g ow h a e was Ul a
igida (11.03 %d-1) ollowed by Schizymenia dubyi wi h 1.58 %d-1 and Valonia u icula is
wi h 1.31 %d-1 and, Colpomenia sinuosa dec eased du ing he whole expe imen a ion
pe iod wi h an a e age o –6.09 %d-1 (Figu e 11).
Figu e 11: G ow h a e p esen ed by he 4 species o algae e alua ed du ing h ee weeks,
s anda d de ia ion and a e age a e p esen ed.
15.93
11.80
5.37
11.03
0
2
4
6
8
10
12
14
16
18
20
Week 1 Week 2 Week 3 a e age
G ow h a e (% d-1)
Ul a igida
2.32
1.60
0.83
1.58
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
Week 1 Week 2 Week 3 a e age
G ow h a e (% d-1)
Schizymenia dubyi
3.46
0.59
-0.12
1.31
-3
-2
-1
0
1
2
3
4
5
Week 1 Week 2 Week 3 a e age
G ow h a e (% d-1)
Valonia u icula is
-5.01
-7.44
-5.83 -6.09
-14
-12
-10
-8
-6
-4
-2
0
Week 1 Week 2 Week 3 a e age
G ow h a e (% d-1)
Colpomenia sinuosa
30
Algae p oduc ion p esen ed a simila endency as he one showed by he g ow h a e,
wi h he highes p oduc ion shown by U. igida ollowed by S. dubyi and V. u icula is
( igu e 12)
Figu e 12: P oduc ion a e p esen ed by he 4 species o algae e alua ed du ing h ee
weeks, s anda d de ia ion and a e age a e p esen ed.
4.3. Algae bio il a ion:
Tables a e p esen ed o summa ize he algae bio il a ion alues o Ni ogen up ake a e
and Ni ogen up ake e iciency, he Ni ogen up ake e iciency was he highes o Ul a
igida wi h 90% howe e , he highes ni ogen up ake a io was shown by Schizymenia
dubyi wi h 0.94 mmol NH4+ m-2d-1, on he o he hand, he lowes o hese pa ame e s was
shown by Valonia u icula is. Each alue p esen ed is an a e age o he iplica es o
each week ( ables 4 o 8).
57.23
35.75
14.78
35.92
0
10
20
30
40
50
60
70
80
Week 1 Week 2 Week 3 a e age
P oduc ion a e (g PS m-2d-1)
Ul a igida
-42.93
-63.79
-50.00 -52.24
-120
-100
-80
-60
-40
-20
0
Week 1 Week 2 Week 3 a e age
P oduc ion a e (g PS m-2d-1)
Colpomenia sinuosa
4.41
3.46
1.94
3.27
0
1
2
3
4
5
6
7
8
9
Week 1 Week 2 Week 3 a e age
P oduc ion a e (g PS m-2d-1)
Schizymenia dubyi
4.60
0.90
-0.19
1.77
-6
-4
-2
0
2
4
6
Week 1 Week 2 Week 3 a e age
P oduc ion a e (g PS m-2d-1)
Valonia u icula is
31
Table 4: Weekly esul s summa izing he a e age amoun o ammonia a he en ance o
he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e o Ul a
igida du ing he expe imen a ion pe iod wi h a densi y o 2g/L and a eno a ion a e o
10 Vol/day.
Ul a ígida
ENTRANCE NH4+
NUE
NUR
GROWTH
RATE
PRODUCTION RATE
WEEK
μM
%
mmol NH4+
m-2d-1
% d-1
g PS m-2d-1
1
3.501
99.999
0.656
15.930
57.225
2
10.380
70.570
1.373
11.800
35.75
3
0.941
99.999
0.176
5.368
14.775
AVERAGE
4.941
90.190
0.735
11.032
35.916
SD
3.985
13.873
0.492
4.345
17.330
Table 5: Weekly esul s summa izing he a e age amoun o ammonia a he en ance o
he anks, he NUE, he NUR, oge he wi h he g ow h and p oduc ion a e o
Colpomenia sinuosa du ing he expe imen a ion pe iod wi h an a e age densi y o
5.76g/L and a eno a ion a e o 10 Vol/day.
Colpomenia sinuosa
ENTRANCE NH4+
NUE
NUR
GROWTH
RATE
PRODUCTION RATE
WEEK
μM
%
mmol NH4+
m-2d-1
% d-1
g PS m-2d-1
1
4.434
87.931
0.731
-3.359
-42.928
2
7.571
74.842
1.062
-4.714
-63.785
3
0.747
99.999
0.140
-4.787
-50.000
AVERAGE
4.251
87.591
0.644
-4.287
-52.238
SD
2.789
10.273
0.381
0.656
8.660
38
Figu e 17: A e age amoun o ammonia a he en ance and exi o each week measu ed
a he en ance and exi o he sea cucumbe anks and i s di e ence.
4.4.3. Sedimen il a ion by Holo hu ia sanc o i:
The amoun s o o ganic ma e and o al ni ogen in he sedimen s a e p esen ed in he
nex igu es, he esul s show an inc ease o bo h o he pa ame e s a he beginning o
each week, when he sedimen s om he sedimen a ion ank we e added, and hen, a e
a week o ac i i y by he sea cucumbe s, he quan i ies o bo h pa ame e s showed a
dec ease, howe e , his dec ease is no signi ican o O ganic ma e (U=25, P=0.18) bu
in he case o o al ni ogen in he sedimen s he di e ence is signi ican (U= 3, P=0.001)
and i s amoun is educed o he con ol le els a he beginning o he expe imen and
does no di e signi ican ly om hem (U=38, P=0.85) ( igu es 18 and 19).
0
1
2
3
4
5
6
7
8
9
10
1 2 3 4 5 6 a e age
Ammonia (µM)
Week
en ance
exi
di e ence

39
Figu e 18: Amoun o o ganic ma e (g/100g) p esen a he beginning and he end o
each week du ing he s udy.
Figu e 19: To al ni ogen (mgN/100g) p esen in he sedimen s a he beginning and he
end o each week o ea men .
2.21 2.22
2.69 2.32
6.08
4.68
3.29
2.83
4.02
3.27
0
1
2
3
4
5
6
7
con ol anks Ini ial Final Ini ial Final Ini ial Final Ini ial Final
s a Week 1 Week 2 Week 3 A e age
O ganig ma e (g/100g)
15.39 13.85
28.54
9.14
24.58
17.06
25.39
14.64
26.17
13.61
0
5
10
15
20
25
30
35
40
con ol anks ini ial inal ini ial inal ini ial inal ini ial inal
s a Week 1 week 2 week 3 a e age
To al ni ogen (mgN/100g)
40
4.4.4. Sea cucumbe g ow h and su i al:
Rega ding sea cucumbe g ow h, a e a pe iod o 100 days o eeding wi h sedimen s
om he sedimen a ion ank e e y 15 days in a e age, he a e age weigh showed a
endency o dec ease in all o he anks including he con ol howe e , he di e ences a e
no signi ican (U=2, P=0.11) (Figu e 20). Su i al in each ea men ank was 100 % while
in he con ol ank only 84.615 % (Table 12)
Figu e 20: Shows he di e ence in he weigh o Holo hu ia sanc o i o each ank a e
100 days o eeding wi h sedimen om he sedimen a ion ank.
Table 12: Su i al o Holo hu ia sanc o i in he expe imen a ion anks and he con ol
ank.
Tank
Su i al (%)
1
100
2
100
3
100
con ol
84.6
A e age
96.15
76.69 76.31 78.38
86.96
79.58
68.19 69.38 67.25
76.95
70.44
-10
10
30
50
70
90
110
130
1 2 3 con ol a e age
weigh (g)
Tank
ini ial
a e 100 days
41
4.5. Rope cul u e expe imen
Rega ding he expe imen o ope cul u e in he bio il e ins alla ion a he PCT, a e 40
days o placing he opes in he bio il e he endency showed a ma ked dec ease o
Hyd opun ia co nea, a sligh inc ease o G a eloupia u u u u and some dec ease o G.
imb ica a ( igu es 21, 22 and 23).
Figu e 21: A e age weigh o Hyd opun ia co nea, G a eloupia u u u u and G. imb ica a
a he beginning o he ope expe imen and a e 40 days.
Figu e 22: A e age g ow h a e o Hyd opun ia co nea, G a eloupia u u u u and G.
imb ica a a he beginning o he ope expe imen and a e 40 days.
100
7
20
12.27 8.45 12.40
0
20
40
60
80
100
120
Hyd opun ia co nea G a eloupia u u u u G a eloupia imb ica a
A e age weigh o algae (g)
Begining
End
-10
-8
-6
-4
-2
0
2
4
Hyd opun ia co nea G a eloupia u u u u G a eloupia imb ica a
G ow h a e (%d-1)
42
Figu e 23: ini ial and inal s a e a e 40 days o he opes s ocked wi h G acilla ia co nea,
G a eloupia imb ica a and G a eloupia u u u u a he bio il e o he PCT.
INITIAL
FINAL
43
5. DISCUSSION
5.1. Ini ial s a e o he e luen s and associa ed o ganisms.
The e was a di e ence in he main algae g oups p esen a he acili ies o he
complimen a y building 1 o he PCT whe e g een and ed algae domina ed and he
acili ies o he PCT whe e b own algae domina ed wi h occasional ou bu s o g een
algae, his is possibly due o he di e ences in he design o bo h ins alla ions, he
complimen a y building 1 o he PCT one is design as a la ge sedimen a ion ank, while
he bio il e a he PCT was o iginally concei ed wi h he aim o using i as a bio il e and
also as an a chi ec onic asse , his means ha he wa e low is conside ably la ge a he
PCT and also ha he su ace a ailable o algae ha ypically g ow a ached is la ge ,
apa om his, he acili ies a he complimen a y building 1 o he PCT a e olde han
he PCT ones so he e has been mo e ime he e o he de elopmen o a mo e di e se
communi y.
The auna assemblage a bo h o he acili ies was domina ed in appa en
abundance by suspensi o es such as Aip asia mu abilis, and less abundan , bu equally
common, sponges and unica es which indica e ha he amoun o nu i ional pa icles in
suspension is high a bo h a eas.
A e y impo an aunis ic componen due o i s di e si y and abundance a bo h
acili ies we e he gas opods and he ac ha mos o hem we e mac oalgui o es (wi h
he excep ion o Ce i hium ulga um) c ea es impo an changes in possible managemen
o hese ins alla ions as algae bio il e s because hey a e expec ed o ea he algae ha
was mean o be used as bio il e s and could suppose a big p oblem o p oduc ion,
howe e , some o hese na u ally g owing o ganisms a e consumed and could ha e
po en ial o aquacul u e, Pa ella aspe a and Osilinus a a us a e examples o his.
The p esence o some species can be indica o s ha some p ac ices in he
aquacul u e acili ies could be imp o ed, o example, in na u al condi ions i would be
ex emely a e o ind Halio is ube cula a coccinea a he e luen s, mo e impo an ly,
he p esence o Dicen a chus lab ax, Spa us au a a and Se iola i oliana a he e luen s
mean ha u he measu es should be aken o p e en escapes.
The modi ica ions made o he di ec ion o he low o he e luen s a he PCT is
expec ed o change in some ways he auna and lo a composi ion in he bio il e , i
would c ea e a be e disposal o he sedimen s hus making hem easie o manage

44
ei he by adding o ganisms able o use hem as eed o by aking hem ou o he bio il e
o be disposed elsewhe e.
Some selec ions had o be made o all he algae and animals ound a bo h
acili ies o u he esea ch, hese o ganisms we e selec ed based on a ious c i e ia, o
example, in he case o Schizymenia dubyi he main in e es was due o i s mo phology,
secondly because i is a ep esen a i e o ed algae which has been ci ed as one o he
p e e ed g oups o abalone o ea in he Cana y Island in wild condi ions (Espino and
He e a, 2002 ci ed in Vie a e al., 2005), and hi dly because o i s abundance a he
sedimen a ion ank o he complimen a y building 1 o he PCT and o i being a species
ha has no been ci ed o he Cana y Islands (Ha oun e al. 2002), on he o he hand, o
Colpomenia sinuosa he main deciding ac o was i s abundance a he PCT, Ul a igida
was abundan and a p o en bio il e o ganism, and Valonia u icula is p esen ed
ela i ely good g ow h.
Fo he il a ion o sedimen s, he sea cucumbe Holo hu ia sanc o i was he only
species p esen ha i he c i e ia o ha ing po en ial as a sedimen eede and as a
ma ke able o ganism, and e en hough some polychae e wo ms we e abundan , he
b oad dis ibu ion and abundance o H. sanc o i (Na a o e al. 2013), i s po en ial
comme cial alue (Aydin. 2008) and i s easy handling we e he de e minan ac o s o
he selec ion o his species
5.2. Algae g ow h and bio il a ion
The g ow h a es di e ed be ween he species as expec ed, being Ul a igida he
species wi h he bigges g ow h a e and p oduc ion, ollowed by Schizymenia dubyi his
concu s wi h a ious e e ences ha men ion ha he genus Ul a is gene ally a e y good
bio il e (Neo i. 2004) and ha one o he mos impo an cha ac e is ics o conside a
species o bio il a ion is he su ace a ea, which inc eases he a ea a ailable o he
up ake o nu ien s
G ow h a e o an algae is la gely de ined by mo phology (Li le and Li le , 1980);
gene ally speaking, he highe he a io o su ace a ea o olume (SA/Vol), he as e he
speci ic g ow h a e. Phy oplank on ha e highe SA/Vol han seaweeds, simila ly, he hin
45
shee mo phology has a highe g ow h a e han does he leshy one ha ha e a g ow h
a e ypically less han 10% day-1 (Ma inho-So iano e al., 2002; Nagle e al., 2003; Neo i.
2004) his explains he ac ha S. dubyi and U. igida had a be e g ow h a e han C.
sinuosa o V. u icula is. Fo hese easons, u he esea ch should be done on he
Schizymenia genus and speci ically in S. dubyi since i ep esen s a possibly iable new
bio il a ion species.
Such la ge s anda d de ia ions indica e ha he expe imen al se ing could be
imp o ed, no necessa ily an ins abili y in he algae cul u es, he g eenhouse se o hese
expe imen s had di e en condi ions o ligh o each ank and he building nex o i
cas ed a shadow upon one o he se s o anks a some hou s, o a oid hese p oblems,
each week anks we e o a ed, howe e , u he measu es should be aken o ensu e
mo e eplicable esul s.
These p oblems could be sol ed inc easing he amoun o eplica es while
changing he disposi ion o he anks ensu ing a mo e e en dis ibu ion o sunligh and
using ligh senso s du ing he s udy a each ank i possible o include ligh and
empe a u e as a co a ia e in he s a is ical analysis.
The a iabili y in he cul u ed species could also be explained by he a iabili y o
he nu ien con en in he e luen s which change acco ding o he ea ed o ganisms a
he acili ies, he hou o hei eeding, he day o he week (no eeding on Sundays) and
o he ac o s such as he cleaning o he acili ies wi h o malin and chlo ine.
The g ow h a e and p oduc ion o Colpomenia sinuosa was nega i e, p obably
because i could no adap o he ee loa ing condi ions; he empe a u e and
pho ope iod a e no expec ed o ha e an impo an ole in hese esul s because du ing
he whole expe imen i was g owing na u ally a he bio il e o he PCT, unde ank
condi ions i usually changed colo om g een o b ownish and hen agmen ed and
s a ed o ge epiphy es
Speci ically in one o he weeks du ing he cul u e o S. dubyi and Valonia
u icula is a b oad cleaning wi h chlo ine and o malin was done in one o he a eas
es ablished o la al ea ing, hese chemicals go o he sedimen a ion ank and could
ha e go en o he algae cul u e anks in high amoun s.
Bio il a ion exp essed by NUR and NUE also p esen ed he expec ed esul s wi h
U. igida and S. dubyi pe o ming good a bo h pa ame e s, Colpomenia sinuosa also
46
p esen ed a e y high ni ogen up ake e iciency e en hough i did no pe o m well
unde he cul u e condi ions, his could mean ha he epiphy es ha g ew on he
Colpomenia a e he eal esponsible o he bio il a ion o ha some bac e ial
bio il a ion could be occu ing.
To u he imp o e hese esul s and he o e all heal h o he algae cul u es some
op ions o p e en ing and comba epiphy es can be applied such as il e ed and e en UV-
ea ed in low wa e which has been ci ed o g ea ly educe he po en ial o epiphy e
con amina ion o a sys em, also, he con ol o e en i onmen al condi ions in he anks
may be used s a egically o in luence compe i ion be ween ma icul u ed seaweeds and
epiphy es, using nu ien s pulses and a high s ocking densi y ha e also been success ully
p o en an i epiphy es echniques (F iedlande e al., 1987, 1991; Neo i. 2004).
The p oposed calcula ion o NUR/ Kg indica es ha , apa om Ul a ( ha may be
g ea ly unde es ima ed due o he li le amoun o ammonia p esen du ing he weeks o
s udy o his species), he species wi h mo e po en ial o bio il a ion and p oduc ion is
S.dubyi. howe e , his calcula ion has some limi a ions because i depends on he amoun
o ammonia a he en ance and he exi o he bio il e which a ied each week and
a ies e en acco ding o he hou o he day; o eally e alua e he po en ial o di e en
species a he same ime he amoun o ammonia should be con olled and he s ocking
densi y o each species should be he same, as well as he amoun o ligh and i s
physiological s a e.
Ano he impo an pa ame e o be con olled is ligh , du ing he weeks o s udy,
i a ied due o cloudiness and logically, I is impo an o men ion ha he maximal
bio il a ion o any nu ien occu s, in day ime, ye some TAN is also aken up a nigh ,
pa icula ly when he TAN load o he algae is low (Cohen and Neo i, 1991; Schuenho e
al., 2003) such as he condi ions o he complimen a y building 1 o he PCT.
5.3. In eg a ed cul u e o Holo hu ia sanc o i and Hyd opun ia co nea
The e luen s om he sea cucumbe anks p esen ed a signi ican ly la ge
concen a ion o o al ammonia his means ha sea cucumbe s ha e he abili y o
eleasing nu ien s om he sedimen in o he wa e column, he amoun o ammonia
47
om hese e luen s was also less a iable han he e luen s om he sedimen a ion ank
which would make i easie o es ima e he p oduc ion o he whole sys em.
The g ow h and p oduc ion a e o H. co nea had a endency o inc ease when
sedimen s and sea cucumbe s we e added o he se up and, e en hough his di e ence
was no signi ica i e, i is expec ed, and he magni ude o he e ec may inc ease wi h
inc eased amoun s o nu ien s being eleased by he sea cucumbe s om he sedimen s,
he la ge a iabili y be ween anks can be explained by di e ences in ligh because o he
o ien a ion o he ank se s a he g eenhouse.
As o he NUE and he NUR he esul s had no signi ican di e ences, his means
ha bo h pa ame e s depend mo e on he species o algae han on he amoun o
ammonia being added and hus, being he same species hese pa ame e s do no change.
On he o he hand, he F /Fm indica es ha he ea men is no in e e ing wi h
he physiological s a e o he algae and hey seem o be well adap ed o bo h cases.
The esul s o o ganic ma e a he beginning o each week o expe imen a ion
whe e e y simila o hose encoun e ed by Lucia Molina in 2000 di ec ly unde he
aquacul u e cages loca ed a Melena a bay, in G an Cana ia.
O ganic ma e p esen ed high a ia ions be ween weeks and no big di e ences
a he beginning and he end o each week; his could be associa ed wi h he e o o he
me hod o es ima ing his pa ame e , which may a y i he sample was no p ope ly
d ied o a cons an weigh due o he humidi y in i o i di e en scales whe e used o
weigh i .
Fo o al ni ogen in he sedimen s he e was a ma ked di e ence be ween he
beginning and he end o each week, especially a he i s week o he expe imen he
di e ence be ween bo h da es was la ge , p obably because o an inc eased eeding
ac i i y by H. sanc o i due o p e ious s a a ion du ing i s ime in qua an ine.
The o al educ ion o ni ogen in he sedimen s does no di e signi ican ly om
he con ol o he beginning o each ank be o e he expe imen ook place, which was
clean sand his means ha he sea cucumbe s whe e e icien in il e ing he ni ogen
om he sedimen s.
The simila i y o he alues o o ganic ma e and ni ogen wi h he ones
encoun e ed below cages in Melena a whe e Molina in 2000 epo ed alues o a ound 4
o 10 % o o ganic ma e di ec ly unde he aquacul u e cages, and be ween 10 – 18
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9. ANNEXES
Annex 1: Example o he iden i ica ion p ocess o Schizymenia dubyi using i s mac oscopic
and mic oscopic mo phology. A) mo phology o he hallus wi h he e y small s ipe B)
Mo phology o hallus in na u al condi ions, C) C oss sec ion o he blade showing
epiphy es ( igh a ow) and medula ilamen s (le a ow), D) C oss sec ion o he blade
showing ea ly de elopmen o he gonimoblas (le a ow), E) c oss sec ion showing he
os iole in a well-de eloped gonimoblas and he expulsion o spo es (le a ow), F) de ail
o he spo es (le a ow).
A
D
B
E
C
F
62
Annex 2: P oposed mechanism o inc ease he su ace a ailable o algae g ow h a he
bio il e o he PCT A) ini ial s a e, B) condi ion a e a mon h iewed om he su ace and
C) condi ion a e a mon h illed wi h Colpomenia sinuosa a he con inuously subme ged
side.
A
C
B