This pape no o be ci ed wi hou p io e e ence o he au ho s
In e na ional Council o he
C.M.
1992/L:21
Explo a ion o he Sea Session V
Zooplank on oxygen consump ion and ñu ien eléase in ela ion
o species composi ion, animáis size and en i onmen al condi ions
in he Bal ic Sea du ing May and Augus
by
L. Pos eU), S. He nández Leon^), M. Gomez^, S. To es^),
U. Mikka l),and A. Po illo Hahne eldl)
1) Ins i u ii Os see o schung (10W) an de Uni e si a Ros ock,
Sees . 15, D-O-2530 Wamemünde, Ge many
2) Facul ad de Ciencias del Ma , Ap do. 550, Las Palmas de G.C.,
Cana y Islands, Spain
KEY WORDS: zooplank on oxygen consump ion, ñu ien eléase, size classes,
composi ion, en i onmen al condi ions, Bal ic Sea, sp ing 1988, 1990, 1991, somme
1988,1990
ABSTRACT:
Zooplank on me abolism in e ms o oxygen consump ion and ñu ien eléase
(ammonia, phospha e) we e measiu'ed in he Bal ic Sea, a empé a e á ea wi h high
en i o mien al changes bo h in space and in ime.
Plank on o he su ace laye we e analysed wi h balance measu emen s in 4 size
classes be ween 50 and 1000 nm du ing sp ing in 1988, 1990 and 1991, in summe
19^8 and 1990 as well.
The use o elec ón anspo sys em (ETS), and he Glu ama e Dehyd ogenase (GDH)
ac i i y as indica o s o espi a ion and ammonia eléase espec i ely, enla ged he
da a densi y and made a h ee dimensional esolu ion a ailable (May 1990, 1991).
Da a a e in he ange o he la i udinal dependend magni ude. They e lec sligh
in e annual, mo e seasonal and egional aspec s. Animáis size, empe a u e, ood
concen a ion, and species composi ion in luence he speci ic a es.
METHODS:
Oxygen consump ion, in eqi e ed as espi a ion, and phospha e and ammonia eléase
(exc e íon) we e measu ed in he Bal ic Sea, mos ly om Mecklenbu g Bigh o he
no hem Go land Sea, some imes om he ansi i ion á ea (Skage ak / Ka ega ) o
he Gul o Finland (Fig.l). De e mina ion we e ca ied ou by means o balance o
"bo le" me hode (Omo i and
Dceda,
1984).
Samples we e ca e iíl coUec ed m he su ace laye down o he he mocline using a
WP-2-ne , equiped wi h a special cod end o p e en , ha animáis ge d y.
A e da k adap a ion o abou
3
o 5 hou s in máximum, unde oxygen sa u a ed
condi ions, using wa e
om
he same s a ion, he plank on was
ac iona ed
in 4 size
classes (55 - 100, 100 - 200, 200 - 500, 500 - 1000
^im),
washed ca e uUy wi h
íl e ed seawa e o clean i
om
nu ien pa ióles, and ans e ed in
11
bo les.
Bo les we e incúba e now in a con aine , cooled by sea su ace wa e , a a o a ing
wheel (2. 5 pm). A leas one con ol bo le wi h he same íl e ed, and oxygen
sa u a ed seawa e is o add, o be able o calcúla e he di e ence o oxygen and
nu ien s o he bo les wi h and wi hou zooplank on a e he 6 o
15
hou s las ing
incuba ion pe iode. The en ichmen o
animáis
in compa ison o in si u condi ions is
abou 20 o 1000 imes, depending om he empe a u e.
Finally oxygen, and nu ien s will be de e mined, he zooplank on s o ed in bu e ed
o maline, o analyse he species composi ion, he d y mass, using con e sión ac o s
as ecommended by BMB (1985). D y mass is needed o calcúla e he speci íc
me abolic a es.
Fo he calcula ion o
in
si u a es he in si u biomass is o de e mine in he abo e
men ioned size classes. To be su e, ha all he di e en size classes a e quan i a i ely
collec ed, ne s o
55
jim, 100 ^m and 200 ^m mesh size we e used o ha pu pose .
F om he 200 jim ne he wo la ges
ac ions
we e p oduced.
Wi h he same ne s plank on o he de e mina ion o me abolic a es by means o
enzyma ic ac i i y
(ETS,
GDH) we e ca ched in
3
dep h le éis (su ace - he mocline,
he mocline - halocline, halocline -
bo om).
Size
ac iona ion
and s o age in liquid
ni ogen we e done immedia ely.
ETS means Elec on T anspo Sys em ac i i y and is in ela ion o espi a ion,
GDH means Glu ama e Dehyd ogenase, o be in ela ion o ammonium eléase.
The single s eps a e desc ibed in a low diag amm (Fig. 2).
The ETS assay was unned in he labo a o y acco ding o Packa d (1969, 1971),
Owens and King (1975) and modi íed by Kenne and Ahmed (1975). The samples
we e homogenized in e lon-glass a
O
- 4
C
using a phospha e bu e
0.1
M, pH = 8
con aining T i ón X-100 (Sigma Chem. Co.) o he solubiliza ion o he enzymes
(Owens and King, 1975). An aliquo o he c ude homogena e was incuba ed o 20
minu es a 12 •€ in da kness condi ions in he p esence o NADH, NADPH, succina e
and an a i icial elec ón accep o , he e azolium sal INT (Biomedical Lab.). A e
his ime, he eac ion is s opped wi h a solu ion con aining phospho ic acid and
o maline and cen i ugued be ween
O
- 4 -C o 10 minu es a 4000 pm. The eac ion
colou was measu ed a 490 nm wi h a u bidi y base line a 750 nm, and espec o a
blank wi hou subs a es and ea ed as he sample. Fo he INT coe icien ac o , he
alué o 1.42 was aken i om he mola abso p ion coe icien ob ained om Kenne
and Ahmed (1975). Final ac i i y alúes we e ecalcula ed o he "in si u"
empe a u e using he A henius equa ion and an ac i a ion ene gy o 15 Kcal/mol.-C
(Packa d e al., 1975). Biomass as p o eins was de e mined acco ding o he me hod o
Low y e al. (1951), using Bo ine Se o Albumin (BSA, Sigma Chem. Co.) as
s anda d. The modi ica ion o he Low y me hod by Pe e son (1977, 1983) was used
o he samples wi h low p o ein con en .
The GDH assay was unned in acco dance wi h Bidiga e and King (1981) and
Bidiga e e al. (1982), including ce ain modi ica ions: One millili e o c ude
homogena e (same as ETS assay) is dilu ed in T is bu e , pH = 8.6 and cen i íigued
(O - 4 -C) o 10 minu es a 4000 pm. In a Pe kin-Elme 551-S UVA^S
spec opho ome e equipped wi h an 1 cm wa e -jacke ed cu e e he mos a ized a 12
•C,
an aliquo o 0.5 mi o he supema an is placed in he p esence o NAD and ADP.
When abso bance lec u e a 340 nm is s able, he assay is unned by adding glu ama e,
and changes in abso bance we e eco ded con inuously o 2 minu es. The slope is
p opo ional o GDH ac i i y. To al olume in he cu e e was 3.0 mi and NAD, ADP
and glu ama e concen a ions we e 1.2, 2 and 40 mM, espec i ely (Bidiga e e al.,
1982).
RESULTS:
1.
Me hodological aspee s:
In gene al he e is a body mass - empe a u e dependency o me abolic a es. A
coincidence wi h calcula ed a es in magni ude should be a quali y ma k o he own
measu emen s, because 700 o 1200 obse a ions a e he basis o he Ikeda's (1985)
equa ions.
Using May and Augus 1988 as an example a sa is ying co espondence is o
obse e:
May Augus
in i o
empe a u e
spec.
oxygen
consump íon
pe indi iduum
Hm^ O2/ Ind.* h
spec.
PO4
eléase pe
indi iduum
pM / Ind. * h
spec.
NH4
eléase pe
indi iduum
pM / Ind. * h
obse a ion
calcula ion
obse a ion
calcula ion
obse a ion
calcula ion
7.50 c
24.64 +/- 10.8;
N=25
10.82 +/- 2.07;
15.17 +/- 7.76;
N=27
4.85 +/- 1.22;
105.32 +/- 92.32;
N=24
70.71 +/- 16.54;
18.3°C
15.06 +/- 7.76;
N=16
17.99 +/. 1.67;
6.90 +/- 6.15;
6.25+/-
0.41;
57.94 +/- 50.38;
N=14
112.57 4-/- 11.16;
In May, a e o du ing he sp ing bloom pe iode, he obse ed alúes a e highe , han
he calcula ed. In Augus he e is he opposi case, indica ing, ha empe a u e is one
in luence, bu he nu i i e basis p obably o highe p io i y.
Nex able includes esul s o co ela ions o 6 da a g oups, he compa isons
(1) o oxygen consump ion and nu ien s eléase on he aw da ale el,
(2) o ETS - and GDH - ac i i y on he aw da a le el,
(3) o d y mass speci ic oxygen consump ion and nu ien s eléase a es ( aw
da a de ided by biomass),
(4) o p o ein speci ícenzyma ic ac i i y,
(5) o (1) and (2),
(6) o (3) and (4),
using he May '91 da a as he basis:
aw da a:
02 s. P04
02 s. NH4
P04 VSNH4
aw da a:
ETS s. GDH
d y mass
speci ic a es:
O2 s. PO4
O2 s. NH4
PO4 s. NH4
p o ein speci ic
ETS s. GDH
aw da a:
O2 s. ETS
NH4VS.
GDH
biomass
speci ic
O2 s. ETS
NH4VS.
GDH
= 0.8309
= 0.9475
= 0.9035
= 0.6730
= 0.6582
= 0.4904
= 0. 5425
= -0.0577
= -0.0476
= 0.3353
= -0.0225
= -0.0937
p<0.00l
p< 0.001
p< 0.001
p< 0.001
p< 0.001
p < 0.05
p < 0.05
N=21
N=21
N=21
N = 23
N = 23
N = 21
N = 21
N = 21
N = 21
N = 21
N = 21
N = 21
Co ela ions we e o expec in all he cases. The signi ican coincidence be ween he
aw da a is a sa is ying indica ion o hei quali y. This is ae o bo h, he da a o
bo le me hode and hoseo he enzyma ic de e mina ions .
The eason o he lesse co ela ion o he d y mass speci ic a es a e p obably he
calcula ed biomass da a. This is wo se in he case o p o ein speci ic ETS- and GDH-
ac i i ies. No o mide s and is he missing co ela ion be ween he aw oxygen
consump ion and he ETS ac i i y on he same le el. Tha biomass speci ic a es o
bo h g oups in he las wo ows don' ma ch does inally no wounde . So he link
be ween he wo me hods, shown in Figu e 2, is no ac i e now. Bo h da a se s ha e
o be sepe a ely used.

2.
Size classes:
The smalle o ganisms show an abou 7 imes highe espi a ion a e compa ed wi h
he la ges g oup (F¡g.3). The mean alúes a e om Augus 1990. The columns
indíca e he s anda d de ia ion.
3.
Ve ical s uc u e:
The de e mina ion o he oxygen consump ion and he ammonia eléase in e ms o
en2yma ic ac i i y in all he dep h le éis allows o es íma e he a e age e ical
s uc u e o me abolic a es.. This will be done as ape cen age in
compa ison
o he
su ace laye in May 1990 and 1991. The e we e no ema kable di e encies
be ween he bo h
yea s,
be ween ETS and GDH, especially in he g oup o p o ein
speci íc a es. La ge e ical g adien s a e o obse e in he in si u -
g oup.
The
me abolic a e dec eases om he su ace laye successi e downwa d by
ap oxima ely 30% pe dep h le el. The ETS esul s om 1991 a e p esen ed in he
nex able as an example:
su ace
laye (up o
he mocline)
in e medía e
laye
( he mocline
o halocline)
bo om laye
(halocline o
bo om)
su ace laye
(up o
he mocline)
in e medía e
laye
( he mocline
o halocline)
bo om laye
(halocline o
bo om)
ETS spec.
1000 - 500 ^im
100
143
101
in si u ETS
1000 - 500 ^m
100
61
25
500 - 200 ^m
100
85
69
500 - 200 xm
100
33
17
200 - 100 }im
100
79
80
200 - 100 nm
100
65
31
o al mean
100
102
83
o al mean
100
53
24
4.
Regional Pa e ns:
Figu e 4.1 includes en i onmen al pa ame e s, like empe a u e, salini y, chlo ophyll
a e aged o he su ace laye , abo e he he mocline, and he diy mass o all size
ac ions, he d y mass o he mos impo an class be ween 100 and 200 ^ini size, and
he equency o he main axonomic g oups wi hin his class. The da a a e egional
a e aged, one o en alúes a e he basis.
SK&KG means Skage ak and Ka ega á ea,
MB Mecklenbu g Bigh ,
AS A kona Sea,
BS Bomholm Sea,
S&CG sou hem and cen al Go land Sea, and
NG No he n Go land Sea (c. . Fig.l).
Figu e 4.2 includes biomass speci ic me abolic a es, oxygen consum ion, ammonia
and phospha e eléase, ETS and GDH ac i i y, and he assimila ion numbe (p ima y
p oduc i i y di ided by chlo ophyll concen a ion).
Figu e 4.3 includes he in si u a es espec i ely.The a e ages a e he same like in
Figu e 4.1.
In a he mos cases a end is isible, cha ac e izing he la e sp ing si ua ion in he
Bal ic Sea, wi h lowe empe a u es in he no hem pa , whe e he me abolic ac i e
cen e is loca ed du ing ha ime, May 1991. The seasonal succession seems o be
o ien a ed om he ansi i ion á ea o he Bal ic Sea o he no hem Go land Sea. The
in si u oxygen consump ion and he phospha e eléase show a clea dec easing
endency, in acco dance wi h he biomass pa e ns in Fig.4.1.
5. Seasonal and in e anual a ia ion
To disc ibe hese a iabili y da a om wo seasons, May and Augus , a e a ailable,
om a leas wo yea s.
In Table 1 (Annex) he mean si ua ion is included o
in si u d y mass, incuba ion empe a u e, in si u equency o Copepods and o
Cladoce ans, he d y mass speci ic and he zooplank on in si u a es o espi a ion
and emine alisa ion, pa ly o di e en size classes.
Compa ing he May si ua ion 1990 was signi ican wa me , han 1988 and 1991.
Also he equency o Copepods / Cladoce ans is di e en . The biomass o he la ge
o ganisms is mo e de eloped in 1990, han in he o he wo yea s. The seasonal cycle
s a ed ea lie in ha yea . This has no signi ícan in luence on he biomass speci ic
me abolic a es, i hey a e a e aged o he whole Bal ic. Bu i has an inpu on he in
si u a es, in dependency o he biomass.
Rema kable di e ences a e o obse e in he biomass speci ic a es, i May and
Augus will be compa ed. Du ing Augus he plank on composi ion is qui di e en
in compa ison o May, he empe a u e. also.
8
I is o conclude, ha seasonal a iabili y is mo e signi ícan han in e annual.
Tempe a u e may ha e an e ec , bu nu i ion and species composi ion o zooplank on
is also impo an . Changes be ween 8 o 12_C (May 88, 91 / May 90) ha e a sligh
in luence o he speci íc a es. A compa ison wi h da a collec ed in 1989 in he
sub opical A lan ic show mo e signi ícan di e ences, bu on he basis o a 10 K
empe a u e change! In ha case a speci íc oxygen consump ion a e o abou 13 mm^
02 /mg d y mass * h in he A lan ic is o compa e wi h 5.55 om Augus
88.
This is
a eIa iono 2.3.
(F om Ikeda (1985) a ac o o 2.8 is o expec , i an á ea o 21° N (A lan ic) and
ano he o 50° N (Bali e Sea) will be compa ed. )
LITERATURE CITED:
BIDIGARE, R.R. and F.D. KING, 1981. The measu emen o glu ama e
dehyd ogenase ac i i y in P aunus lexuosus and i s ole in he egula ion o
ammonium ex e ion. Comp. Biochem. Physiol., 70 B: 409 - 413.
BIDIGARE, R.R., F.D. KING and D.C. BIGGS, 1982. Glu ama e dehyd ogenase
(GDH) and espi a o y elec on- anspo -sys em (ETS) ac i i ies in Gul o México
zooplank on. J. Plank on Res., 4: 895 - 911.
HERNROTH, L. & H. VIL JAMA A (Eds.), 1979. Mesozooplank on biomass
assessmen . BMB Publ. No. 6 . I5pp.
IKEDA, T., 1985. Me abolic a es o epipelagic ma ine zooplank on as a imc ion o
body mass and empe a u e. Ma . Biol.. 85: 1 - 11.
KENNER, R.A. & S.I. AHMED, 1975a. Measu emen s o elec ón anspo
ac i i ies in ma ine phy oplank on. Ma . Biol.. 33: 119 - 127.
LOWRY, P.H., N.J. ROSENBROUGH, A.L. FARR & R.J. RANDALL, 1951.
P o ein measu emen wi h a Folin phenol eagen . J. Biol. Chem.. 193: 265 - 275.
OMORI, M. & T. IKEDA, 1984. Me hods in ma ine zooplank on ecology. John
Wiley & Sons. New Yo k, Chiches e , B isbane, To on o, Singapo e. 332 pp.
OWENS, T.G. & F.D. KING, 1975. The measu emen o espi a o y elec ón
anspo sys em ac i i y in ma ine zooplank on. Ma . Biol., 30: 27 - 36.
PACKARD, T.T., 1969. The es ima ion o he oxygen u iliza ion a e in seawa e
om he ac i i y o he espi a o y elec ón anspo sys em in plank on. Ph. D.
Thesis, Uni . Washing on, Sea ie. 115 pp.
PACKARD, T.T,,
1971.
The measu emen o espi a o y elec ón anspo ac i i y in
ma ine phy oplank on. J. Ma . Res., 29: 235 - 244.
Enz mology. Vol.
91.
Academic P ess, pp
95
- 119.
ACKNOWLEDGMENT:
We would like o hank he c ew o
RV
"A. .Humbold ", o
RV
"P o .A.Penck" o
hei
help,
he Chemical Depa men o he Ins i u e o
Bal ic
Sea Resea ch,
Wamemünde, o many nu ien analyses, Anneli Pos el and Heide Sandbe g o he
nume ous zooplank on de e mina ions, he lOW Da a Cen e, p o iding empe a u e,
salini y, chlo ophyll da a, and assimila ion numbe s, and he BMFT, because he
measu emen o
1991
we e suppo ed by his Minis y (Con ac No.:03F0030A).
N =
mg d y nass-m''
í incuba íoo empe a u e/oC
equency o
Copcpods/
%
equency o Gadoce ans
Oxycen coosomp ion
d y mass !^)eci ic/
mm'
Oi-mK"'-h''
in si u/
mm'
Oj-mg'-h''
',
Pbospha e eléase
d y mass sped íc/
nM-mj-'-í -'
in si u/
:
>íM-m-'-d'
A lmonia eléase
; d y mass speciñd
nM-mg-'-h-'
1
in si u/
;iM-m'-d'
.
Size ac ion//im
May '88
26
27
7,5
76
16
9,12
7,24
5,40
3,50
34,68
42,66
>200
4,19
Í2,T~
43
46
13,93
1.4
4,30
0,43
53,48
5,38
May'90
16
19,50 __ 4.32 _ -
1
4Ú 10 13
41 12 23
4,77 8,95 32,33
2.23 0,93
E - 4.56
3,12 7,65 21,70
1,46 0.87
'-"2,76
16,51 29,76 38,65
7,73 3,08
E » 16,19
1000-
500- 200- 100-
500 200 100 55
>20Ó •" •
May'91
12
4.77 8.59
'"8;6
So 34
11___22__
8.16 5,18
0,93 1,07
19,26
3
0
8.20
3,70
E-.6.5
3,78 1.92
0.43 0.40
2,19
1,01
E - 2.03
30,48 17.35
3,49 3,58
22,67
10,48
E = 18.44
1000-
600-
500 200
>20b
200-
__100_
1,61
0
0
18,28
0,71
5.01
0,19
23,06
0,89
100-
_55 _
Aug. '88
16
100
18,3
Mi
70
5.55
__13,37
2,55
5741
21,00
53,61
>200
_J,86
31"
67 ,
5,26
1,29
0,65
0,14
24.66
5.25
1000-
500
65.37
18.2
29
70
6.29
8,48 '
1,49
2.34
20,08
31,50
500-
200
>200
Aug. '90
5
12.51
29
69
11,61
3,98
T m 15 30
2,60
0,79
E - 3,38
15,25
4,64
E - 43.38
200-
lOQ
I 09
41
57
39,68
1,64
4,06
0,11
71,70
1,99
100-
55.
_
Table: Co npa ísons o mean alúes o en i onmen al condi íons. zooplank on
biomass. equency o Copepods and PhyHopods. speci ic and in si u mc abolic a es
in May I988.I990. I99I and in Augus I988.I991.