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S-and N-cy­cling in cold seeps, ani­mal as­so­cia­ted mi­cro­bes, dia­toms and in ma­ri­ne se­di­ments

Lichtschlag, de Beer, Heisterkamp, Stief, Schreiber, Kuipers, Schwermer, Weber, Abed, Allers, Wang, Kamp, Behrend

Anna Licht­schlag in­ves­ti­ga­ted bio­lo­gi­cal and che­mi­cal pa­thways of sul­fi­de oxi­da­ti­on in cold seeps. She found that in the Haa­kon Mos­by Mud Vol­ca­no sul­fi­de oxi­da­ti­on is ent­i­re­ly bio­lo­gi­cal, as the sul­fi­de flux is pro­por­tio­nal to the ni­tra­te uptake by Beggia­toa, and thus leads to pri­ma­ry pro­duc­tion in this in­de­ed rich and di­ver­se ha­bi­tat. Re­mark­bly, in a seep in the an­ae­ro­bic zone of the Black Sea she found sul­fi­de oxi­da­ti­on to sul­fur, dri­ven by ex­tru­ded oxi­di­sed iron. In the mud vol­ca­no high CH4 le­vels have been mi­xed for mil­le­nia with oxi­di­sed Fe(III). This me­ans that mi­cro­bi­al CH4 oxi­da­ti­on by Fe(III) is pro­bab­ly im­pos­si­ble. Only af­ter re­duc­tion of the CH4 by sul­fa­te Fe(III) can be re­du­ced.
Anna Beh­rend aims to elu­ci­da­te what con­trols whe­ther DNRA or de­ni­tri­fi­ca­ti­on to N2 do­mi­na­tes. She star­ted long- and short-term in­cu­ba­ti­on ex­pe­ri­ments with sul­fi­de and man­ga­ne­se ad­di­ti­ons, and will ana­ly­se the mi­cro­bi­al pro­ces­ses and com­mu­ni­ty shifts.

Anja Kamp (with La­vik & Kuy­pers) tested the hy­po­the­sis that dia­toms store ni­tra­te and can use it to re­spi­re in deep se­di­ments. She has se­veral li­nes of evi­dence that this in­de­ed hap­pens. This would ex­plain the ob­ser­va­ti­on that ni­tra­te can be found de­ci­me­ters deep in co­as­tal se­di­ments.

Left: Distribution of nitrate from microsensor data (•) and total extraction (bars). The difference is intracellular in diatoms.

Cal­ci­fi­ca­ti­on in ma­ri­ne se­di­ments and freshwa­ter stroma­to­li­tes

Bissett, Schoon, Alexandra Rao, Polerecky, Martin Glas

It is well known that cal­ci­fi­ca­ti­on is cou­p­led to pho­to­syn­the­sis, and that bent­hic pho­to­syn­the­sis can lead to cal­ci­fi­ca­ti­on. BioAcid is a lar­ge Ger­man col­la­bo­ra­ti­ve re­se­arch ef­fort (For­scher­grup­pe) fun­ded by the BMBF, that aims to in­ves­ti­ga­te the ef­fects of oce­an aci­di­fi­ca­ti­on on ma­ri­ne life. It was re­por­ted that cal­ci­fi­ca­ti­on in co­rals and coc­co­li­tho­pho­res is re­du­ced. We fur­ther in­ves­ti­ga­te the hy­po­the­sis that bent­hic cal­ci­fi­ca­ti­on, es­pe­cial­ly of pho­to­tro­phs, is not very sen­si­ti­ve to aci­di­fi­ca­ti­on. The con­side­ra­ti­ons are that the or­ga­nisms crea­te the site of cal­ci­fi­ca­ti­on by me­ta­bo­lic pro­ces­ses, and that trans­port in se­di­ments or the MBL is dif­fu­sio­nal­ly re­stric­ted. In­de­ed mi­cro­sen­sor mea­su­re­ments have shown that the sur­face pH of ac­tive se­di­ments stron­gly de­via­tes from the wa­ter­co­lumn. Al­ban Ra­met­te from the Ha­bi­tat group par­ti­ci­pa­tes with a stu­dy on the ef­fect of ex­tre­me aci­di­fi­ca­ti­on on mi­cro­bi­al com­mu­nities in se­abeds near na­tu­ral CO2 seeps. Also di­ver­si­ty of meio-, ma­cro- and me­gafau­na will be as­ses­sed to ad­dress thres­holds of low pH on bent­hic eco­sys­tems.
Ex­ten­si­ve mea­su­re­ments in reef se­di­ments (Schoon) du­ring diel cy­cles show­ed that the pH va­lue re­mai­ned high du­ring most of the night, and that cal­ci­fi­ca­ti­on is not con­fi­ned to the light pe­ri­od. A stu­dy on a hard­water creek, whe­re cal­ci­fi­ca­ti­on on tu­fas (sort of stroma­to­li­te) was thought to be dri­ven by CO2 out­gas­sing. Howe­ver, in- and ex-situ mi­cro­sen­sor mea­su­re­ments (An­d­rew Bis­sett) show­ed that cal­ci­fi­ca­ti­on on the tufa is ent­i­re­ly con­trol­led by pho­to­syn­the­sis. The out­gas­sing does lead to cal­ci­um car­bo­na­te fall out, but this is wa­s­hed away and does not reach the tufa. The ef­fect of pH of the wa­ter co­lumn on bent­hic cal­ci­fi­ca­ti­on was stu­di­ed in the tu­fas and in reef se­di­ments. In the pH ran­ge of 7.2-8.4 cal­ci­fi­ca­ti­on was not in­flu­en­ced. It was con­clu­ded that the bent­hic mi­cro­bi­al ac­tivi­ty much more stron­gly de­ter­mi­nes the lo­cal pH than the pH of the sea­wa­ter. This is high­ly va­luable in­for­ma­ti­on in view of the on­go­ing aci­di­fi­ca­ti­on of the oce­ans, and the pu­bli­ca­ti­on in AEM got an edi­to­ri­al com­ment.
Ra­phae­la Schoon in­ves­ti­ga­ted whe­ther cal­ci­fi­ca­ti­on in si­li­ca­te and car­bo­na­te se­di­ments are equal, when sea­wa­ter che­mis­try, amount of bent­hic pho­to­tro­phs, pho­to­syn­the­sis ra­tes and il­lu­mi­na­ti­on are iden­ti­cal. Only in car­bo­na­te se­di­ments cal­ci­fi­ca­ti­on could be in­du­ced by il­lu­mi­na­ti­on. This me­ans that cal­ci­fi­ca­ti­on nee­ds cal­ci­te to pro­ceed, pro­bab­ly as nu­cleus.
Alex­an­dra Rao stu­dies cal­fi­ca­ti­on and de­cal­ci­fi­ca­ti­on in per­me­able se­di­ments. A very chal­len­ging task, but she is har­ves­ting va­luable data now, e.g. she mea­su­red the ki­ne­tics of cal­ci­fi­ca­ti­on and cal­ci­um dis­so­lu­ti­on by ra­pid pH per­tur­ba­ti­ons of the po­re­wa­ter. This is pos­si­ble only in per­me­able se­di­ments. Her se­cond tool is mo­de­ling, in col­la­bo­ra­ti­on with Phi­lip Meis­man (NIOO, Nl).

Ima­ging of in­tra­cel­lu­lar pro­ces­ses

Beutler, Hinck, de Beer

Mar­tin Beut­ler and Jana Mi­lucka (Mi­cro­bio­lo­gy) found that the va­cuo­le in Beggia­toa is not a pas­si­ve tank for ni­tra­te, but ge­ne­ra­tes en­er­gy for cel­lu­lar use. Ge­no­mic in­for­ma­ti­on (Marc Muss­mann, Mole­co­lar Eco­lo­gy Group) and a se­ries of cle­ver ex­pe­ri­ments re­vea­led that the va­cuo­le mem­bra­ne has an elec­tron trans­port chain, can re­du­ce ni­tra­te that ge­ne­ra­tes a pro­ton mo­ti­ve force (aci­dic and po­si­ti­ve in­si­de). A com­pa­ri­son with mi­to­chon­dria is mis­pla­ced, the va­cuo­le may have the same func­tions as the ou­ter mem­bra­ne, but is in­ver­ted as an in­va­gi­na­ti­on. Sur­pri­sin­gly, howe­ver, Cyt-c was only found in the va­cuo­le. The va­cuo­le is of cour­se not a cell or­ga­nel­le, it can not re­pli­ca­te, DNA in­si­de the va­cuo­le was not found. It de­mons­tra­tes how com­plex also pro­ka­ryo­tes can be struc­tu­red.
The vacuole as cellular powerplant. Nitrate reduction builds up a PMF (pH and potential difference), that is used to conserve energy in the form of poly-fosfate and ATP.
Immunostaining by antibodies showed that cytochrome c (thus a respiration chain) is present in Beggiatoa vacuoles. A) antibody Texas Red signal. B) reflection signal caused by sulfur globules. C) transmission image. D) overlay of A)-C).

Mass trans­fer and con­ver­si­on in po­rous se­di­ments

Wu, de Beer, Beck, Polerecky, Janssen, Sivelgen, Nemecki

In the ECO­DIS pro­ject we (Beck, Po­le­recky, Jans­sen) in­ves­ti­ga­ted the hy­po­the­sis that mi­cro­bi­al pro­ces­ses can con­trol tra­ce me­tal ex­ch­an­ge bet­ween se­di­ment and wa­ter co­lumn. By a com­bi­na­ti­on of la­bo­ra­to­ry ex­pe­ri­ments and field stu­dies we could cor­re­la­te diel fluc­tua­ti­ons in a stream with bent­hic pho­to­syn­the­sis. The dif­fe­rent be­ha­vi­or of Pb, Cu, Ni and Cd was ex­plai­ned by dif­fe­rent re­s­pon­ses to pH. The work re­sul­ted in a pu­bli­ca­ti­on that was in unusual­ly strong terms prai­sed by the chief edi­tor Jac­ques Buf­fle.
The stu­dy on mass trans­fer pheno­me­na in the in­terti­dal flats re­aches its fi­nal sta­ge. Chris­ty Wu in­ves­ti­ga­tes the de­eper are­as un­der the flats to lo­ca­te the sour­ce of the me­tha­ne that seeps from the flats at low tide. She stu­di­ed the bio­geo­che­mis­try of two deep co­res of 20 m length, in­clu­ding sul­fa­te re­duc­tion and AOM ra­tes. 14C da­ting of the C-org sug­gests ex­ch­an­ge of or­ga­nics and sul­fa­te down to 20 m depth. She also mea­su­red po­re­wa­ter che­mis­try in tran­sects from low wa­ter line to 200 m in­wards. Ins­tead of a lo­cal me­tha­ne plu­me, she found a me­tha­ne – sul­fa­te tran­si­ti­on zone ca 2 m be­low the sur­face ever­yw­he­re. She will fur­ther se­arch for the sour­ce of re­du­cing power.
Lub­os found new pheno­me­na as­so­cia­ted with bio­ir­ri­ga­ti­on ac­tivi­ties of the lug­worm Are­ni­co­la.
It was ge­ne­ral­ly as­su­med that car­bo­na­te se­di­ments are more mi­cro­bi­al­ly ac­tive than com­pa­ra­ble si­li­ca­te sands. This was in­ves­ti­ga­ted in de­tail by 2 MSc stu­dents on Elba, whe­re the two ha­bi­tats are clo­se­ly to­ge­ther. Ra­tes of re­spi­ra­ti­on, pho­to­syn­the­sis and sul­fa­te re­duc­tion were si­mi­lar. No mi­ra­cle with car­bo­na­te sands.
Lo­phe­lia (Abed, Al­lers): CO­RAMM was a Sta­toil-fun­ded pro­ject ai­med to as­ses re­por­ted da­ma­ge by oil ex­plo­ra­ti­on ac­tivi­ties. We in­ves­ti­ga­ted the hy­po­the­sis that da­ma­ge was cau­sed by lo­cal an­oxia and an­ae­ro­bic mi­cro­bi­al pro­ces­ses. Howe­ver, we found that Lo­phe­lia quick­ly re­mo­ves high loads of of se­di­ment or drill cut­tings. Com­ple­te bu­ri­al is sur­vi­ved for 24 hours. Va­rious other li­nes of evi­dence lead to the con­clu­si­on that Lo­phe­lia is far too ro­bust to suf­fer from off shore ac­tivi­ties, un­less they are smas­hed by an­chors or trawls. Com­mu­ni­ty com­po­si­ti­ons from drill cut­tings (es­sen­ti­al­ly deep bio­s­phe­re) and from seaf­loor se­di­ments did not con­ver­ge when ex­po­sed to the same con­di­ti­on. Ines Heis­ter­kamp stu­dies the N2O pro­duc­tion by mi­cro­bi­al com­mu­nities as­so­cia­ted with ma­ri­ne ani­mals. She found that es­pe­cial­ly bio­film co­ve­r­ed ani­mals pro­du­ce high amounts, alt­hough also gut com­mu­nities can be very ac­tive. It is plan­ned to ex­pand the stu­dy to krill, DFG-fun­ding for ship time is re­quested. Sal­in­i­ty de­pen­dence of de­ni­tri­fi­ca­ti­on in bio­films from desa­li­na­ti­on plants and hy­per­sa­li­ne mi­cro­bi­al is stu­di­ed with Is­rae­li part­ners. Com­mu­nities ap­pe­ar to be ad­ap­ted to pre­cise­ly the am­bi­ent sal­in­i­ty. The bio­films are well struc­tu­red la­mi­na­ted gra­nu­les, with ab­un­dant me­tha­no­gens. De­ni­tra­ti­fi­ca­ti­on sti­mu­la­ted me­thyl­a­mi­ne for­ma­ti­on, a sub­stra­te for me­tha­no­gens. Cal­ci­fi­ca­ti­on ap­peared to be dri­ven by ni­tri­te only, ni­tra­te re­duc­tion to ni­tri­te may oc­cur only at the ou­ter shell, or in the wa­ter­pha­se.
In a side pro­ject it was found that also hu­mans pro­du­ce high amounts of N2O, most­ly from den­tal plaque. Ni­tra­te is for­med in the body by NO (neu­ro­trans­mit­ter) oxi­da­ti­on and is con­cen­tra­ted in the sa­li­va (ca 1 mM). Ni­tra­te is re­du­ced in the mouth to ni­tri­te, and due to the aci­di­ty in the plaque (pH<6) and sto­mach, che­mi­cal­ly to NO, and N2O. NO is thought to be es­sen­ti­al in the ste­ri­liza­t­i­on of food in the sto­mach, and be im­portant in fight­ing gum in­fec­tions. Thus in hu­mans, and li­kely in other ani­mals, an unusu­al ni­tro­gen cy­cle re­lea­ses the green­hou­se gas N2O.

Mi­cro­bi­al mats

Kohls, Abed, Polerecky, Al-Najjar, Hausler, Klatt, Chennu

Hy­per Spec­tral ima­ging is now being de­ve­l­o­ped for un­der­wa­ter use, in the EU net­work Sen­se­N­et. Chen­nu/​Po­le­reecky will mea­su­re com­mu­ni­ty dy­na­mics in pho­to­tro­phic mats and on pho­tic se­di­ments, and im­pro­ve vi­deo mo­sai­cing of the deep-seaf­loor. A por­ta­ble lab-sys­tem is cur­rent­ly used in va­rious pro­jects. In a sul­fi­dic stream Po­le­recky, Klatt, Haus­ler stu­dy sul­fi­de oxi­da­ti­on by an­oxy­ge­nic- and oxy­ge­nic pho­to­syn­the­sis, ae­ro­bic and an­ae­ro­bic oxi­da­ti­on by fil­amen­tous bac­te­ria. The ef­fi­ci­en­cy of light con­ver­si­on to bio­lo­gi­cal en­er­gy in 3 dif­fe­rent eco­sys­tems was stu­di­ed by ma­king ba­lan­ced bud­gets: heat, re­flec­tion and pho­to­syn­the­sis. The quan­tum ef­fi­ci­en­cy of mats from 5 dif­fe­rent ha­bi­tats was de­ter­mi­ned by PAM ima­ging, and com­pa­red with bac­te­ri­al di­ver­si­ty mea­su­red by ARI­SA fin­ger­prin­ting. Ka­tha­ri­na Kohls found a no­vel type of mi­gra­ti­on in re­s­pon­se to a sal­in­i­ty gra­di­ent. Ra­eid Abed has made a lar­ge li­bra­ry of fin­ger­prints from sour­ce are­as for dust (salt lakes in the Aus­tra­li­an out­back), he com­pa­red com­mu­nities in dust and could lo­ca­te the sour­ce area. He found that dust can con­tain strains found in Ant­arc­tica, Af­ri­ca and Asia.
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