Influence of ammonium availability on expression of nifD and amtB genes during biostimulation of a U(VI) contaminated aquifer: implications for U(VI) removal and monitoring the metabolic state of Geobacteraceae Page: 4 of 8
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D*Ilher A ai*tiij lionFISKE . (a) Milrebllet omumty copinpidon based uM US MMA dune ueMnces exlramd hom the pumodwater at ie
nw1iludedng locallns baloF nrosat injecias b.man. (hil Ammnimn concmealos observed prior to eefat* injecden inia4 blue
el ard average oncaaraiorn 4mrsn n + stili bf Weation OmropeHidlIS to 165 DNA clute libraries. (el Spelstumporal
ar Sium uctuadiuOm during ne4 a e nmetconcentrationsrelative to acetate crncent rainons suggested
that relatively more acerate had been consruntei an Ilis
location (Figure S1 of the Supparting lnfornistion). As
expected from previous studies (3, 29. acelate amend man i
stimulated dissimilatory metal reductlan a5evidcnccd by
an accumulation of dissolved Fe(1I) (Figure c) and a loss
of iMV) at all four locations (Figure 2a], [(V1) reduc lion
wasstimulated to the lowest concentrationsin D-02, where
some of the higher acetare concentrations were delivered
(Figure 2a).
Signifcantdifferenceswere obsewd in arnmoniunkievels
across the experimental plot, with concentrations varying
by2-ordersofmagnitude between the wells (Figure b) Well
D-0 had significantly higher ammonium levels, wlereas
D-O5 had significantly tower levels than those of all oilier
locations (p-value <0.02) (Figure 3b). While locations D-02
and [.04 showed increases in ammonium during the
amendrent,no significant trends wee observed (Flguxe 3c).
Consistent with theexpecredlack ofnitrateatid nuirlte under
dissimifatory metal-reducing conditions, other sources of
inorganic nitrogen were below detection (<10M) In oil wells
throughout the experiment.
MiLubhlfCommuniltyDynamica Differences in grouid-
water ammonium concentrations may have been uric factor
contribuling to variations in the microbial com unity
composition beforeand duringacetate amendrten. Analysis
of 163 RNAgene sequences extracted frem the groundwalcr
indicated that microorganisms within the ft and 6-proteo-
bacteria classes were among the nostabuidanit prior lo ond
during the i run-reducingphase ofacetate amendment (Pigure
3a). The 165 rONA clone sequences related io Decldwtmorws
species (88-99% similarity) were abundant al all five silos
prior to the addition of acetate to the groundwater (Figure
33 of the Supporting Infotnation) and represeritet inure
than 60% of sequences in D-5, which had ihe lowest
a mmanium levels (Flgure3a).Temporalarnples fl-rn three
locations that spanned low, intermediate, and high am-nlniunj [evels showed a decline in relative abundance of
Dechlorrwaus peCeks following acetate iidlliins by day
nine and a recurrence sfior 18 days in D-08, which had the
highest ammoniunt levels 0(igure SZ of lire Supporting
inforniatiori).
Relative proportions of Geobacer and Rhodofprax
species were comparable at thesules with lower ammoniun
levels priUT to acetale addition, including D-05, U-01, and
D-02 (Figure Sal. However, at Ioca lions with higher initial
ars man inm, including D-04 ani D-08, Rhvd ferusx species
(91-9% similarity) wore more abundant prior tO acetaie
injection (Figure 3a and Vigure 53 of the Supporting
Inforration) but declirnd in relative abundance after a
week of acetate amendment {Figure S2 of Ihe Supporting
Irformnation). As expecied from previous studies (3-6),
Gobacer species (80-9M% similarity) became the pre-
dorninanicommunitymnetmbersafter acstateameridment,
regIrdiess o smrnoni uni cuncentralicniii (Figure 32 of the
SupporIitiginFormation] l-Consistent with studies of other
uranium coniaininated sites (5. 18, 35, Ceones with high
sirrmiriy t1 .Aher f-prot obareria, including Nroswona-
daceae,. Comanfonadaeeae, and Oxaobnwfene family ies
weredetecied in 2-10% of clone sequencespriortoaoetate
a-nendrent (Figume 3a).
in Situ Faprensio of Ammuaoshn Trngnoner and
Nlin u Mixation Goone in GophTer Specles, in order to
leam more about the in Siltu melabolism of subsurface
Gegjtacter species tie nurnber of transcriplis for the am-
Awmor um Irsportergeine, am0, and 1he nitrogen fixation
gene, ni#P was quantified during the bloslimulalioi experi-
rneni. [n general the number of r1D I rairscrpts was slightly
less abundant than ihose for the housekeeping gone revA,
resulting in riedisn nifD arid rgtA levels between 0.28 and
0.43 [Figure 4a). Overall zfD abundance levels were nut
statistically different among the fear monitoring locations
(p-value = 0.43) (Figure a. A previous study (is) demun-
slraied that Cepdcrer species living in sediments have4
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Mouser, Paula J.; N'Guessan, A. Lucie; Elifantz, Hila; Holmes, Dawn E.; Williams, Kenneth H; Wilkins, Michael J. et al. Influence of ammonium availability on expression of nifD and amtB genes during biostimulation of a U(VI) contaminated aquifer: implications for U(VI) removal and monitoring the metabolic state of Geobacteraceae, article, March 25, 2009; Berkeley, California. (https://digital.library.unt.edu/ark:/67531/metadc829484/m1/4/: accessed April 19, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.