Biological Fixed Film - IngentaConnect

Biological Fixed Film Mary Joy Wesley, Robert N. Lerner, Eun Sik Kim, MD Shahinoor Islam, Yang Liu*

ABSTRACT:

Literature published during the year

medical and dental biofilms, corrosion related to biofilms,

2010 on the topic of biological fixed film including biofilm

biofilm formation in pipes and distribution systems,

and bioreactors for the purpose of water and wastewater

biofilms for the treatment of air pollution, and biofilms in

treatment has been reviewed herein. Literature published

the natural environment. Anaerobic biofilms are included in

regarding microbial fuel cells has also been included. The

another review paper and will not be included herein. Organization and Coverage. The sections have

review has been organized into the following sections: biofilm

formation,

adhesion,

quorum

sensing,

and

been divided into research focusing on the biofilm

consortia; techniques and analytic methods; biofilm

formation, adhesion, quorum sensing (QS), and consortia;

reactors and modeling; and microbial fuel cells.

techniques and analytic methods; biofilm reactors including carriers and modeling; and MFCs. Within the reactor

KEYWORDS: Biofilm,

biofilm reactors,

modeling,

microbial fuel cells, and microbial electrolysis cells.

section, modeling specific to a particular reactor type is included in the section pertaining to that particular reactor type. Otherwise, the modeling is included in a general

doi: 10.2175/106143011X13075599869254

modeling section. Membrane reactor research represents a large area of research for the current review year and has

Introduction

been

subdivided

into

fouling,

cleaning,

antifouling

Overview. This review focuses on aerobic

techniques, and modeling. The section pertaining to MFCs

biological fixed film systems used for the purpose of water

has been subdivided into several sections including fuel cell

and wastewater treatment as well as microbial fuel cell

electrodes, characterization of fuel cell biofilm, fuels, mode

(MFC) research. Therefore, it excludes research regarding

of fuel cell operation, fuel cell membranes, microbial electrolysis fuel cells, and other fuel cell research.

————————— *Department

of

Civil

&

Environmental

Engineering,

Markin/CNRL Natural Resources Engineering Facility, University of Alberta, Edmonton, AB T6G 2W2, Canada; *Corresponding author phone:

780-492-5515;

[email protected]

Fax.

780-492-0249;

E-mail:

Biofilm Formation, Structure, and Behavior Biofilm

Formation.

Stenotrophomonas

maltophila that was capable of degrading branched anionic surfactants in activated sludge (AS) was immobilized on

1150 Water Environment Research, Volume 83, Number 10—Copyright © 2011 Water Environment Federation

glass beads and shown to have a 2-fold removal rate for the

The ATP content of granular biomass was found

substrate as compared with a non-immobilized strain

to be related to initiation of microbial aggregation in

(Farzaneh et al., 2010). Theunissen et al. (2010) identified a

granules whereas AI-2 content was shown to be related to

biofilm promoting factor A from Shewanella oneidensis

granule maturation and the coordination of microorganisms

which was secreted by a type I secretion system and found

in the aggregates (Xiong and Liu, 2010b). A cellular

to be modulated by calcium.

metabolic poison, 2,4-dinitrophenol was used to disrupt

Cultivability of some species of bacteria has been

ATP synthesis, leading to a reduced production of ATP-

associated with a requirement for siderophores from other

Mediated Autoinducer-2 (AI-2) and suppression of

species. A new acyl-desferrioxamine siderophore was

attachment (Xu and Liu, 2010). The AI-2 content of the

shown to be the growth factor produced by one helper

microorganisms was found to be positively related to the

strain (D’Onofrio et al., 2010).

extent of attachment.

Dror-Ehre et al. (2010) found that the exposure of

Robledo-Ortiz et al. (2010) showed that the

Pseudomonas aeruginosa to molecularly capped silver

adsorption

(Ag) nanoparticles retarded the formation of biofilm

polyethylene foam composites followed the first order

although the cells were still able to produce extracellular

kinetics and was irreversible. They further studied the

polymeric substances (EPS) and a relatively low adherent

impact on adhesion of factors including pH, contact time,

biofilm.

temperature, initial biomass concentration, and ionic Adhesion. Adhesion mechanisms and modeling

of

P.

putida

onto

agave-fiber/recycled

strength.

were reviewed by Hori and Matsumoto (2010). Cell

Initial cell adhesion of a model biofouling

appendages including polysaccharide chains and proteinous

bacterium P. aeruginosa PAO1 was studied (Myint et al.,

nanofibers were found to be responsible for both specific

2010). Although the initial adhesion was related to the

and nonspecific cell adhesion to surfaces as well as for a

hydrophobicity of the membrane, after 48 hours, the extent

possible source of model deviation. Landini et al. (2010)

of the P. aeruginosa PAO1 biofilm was found to be

reviewed anti-biofouling agents focussing on inhibition of

independent of the surface property. Liang, Das, et al.

QS, cyclic di-GMP metabolism, DNA and nucleotide

(2010) studied the effect of solution ionic strength on the

biosynthesis, and activation of biofilm dispersal. Two

adhesion tendency of P. putida on two different RO

Klebsiella pneumoniae AI-2 transport mutants showed

membranes

increased

formed

fractionation. They showed that high ionic strength resulted

biofilms with impaired architecture but greater biomass (De

in greater adhesion and ionic strength had a greater impact

Araujo et al., 2010).

on adhesion than did membrane hydrophilicity. They also

lipopolysaccharide

production

and

using

asymmetrical

flow

field

flow

found that the impact of ionic strength was more


pronounced for the hydrophilic membranes while the

Sands naphthenic acids (Headley et al., 2010). This

hydrophobic membrane had a greater bacterial adhesion

inability was attributed to three key factors including the

than did the hydrophilic membrane. Zhao et al. (2010)

molecular structure and molecular mass of the naphthenic

grafted copolymer brushes onto the surface of a

acids, and inhibition by sulphur (S) and/or nitrogen (N)

polypropylene

containing acids.

membrane

using

UV-induced

Cold temperature (5C) nitrifiers in a

polymerization of two oppositely charged polymers,

sludge nitrifying lagoon were found to be affiliated with

thereby improving the hydrophilicity of the membrane.

mostly beta-Proteobacteria, some Bacteroidetes, and some

Anti-protein adsorption was assessed with bovine serum

Actinobacteria (Ducey et al., 2010). While the majority of

album and lysozyme by which it was determined that the

the bacteria belonged to the genus Nitrosomonas, some had

more balanced membrane surface charge had the best non-

affiliation with floc forming or psychrotolerant bacteria.

biofouling. Consortia. Wojnowska-Baryla et al. (2010) reviewed bacterial communities in wastewater treatment

Quorum sensing. Choudhary and SchmidtDannert (2010) reviewed QS and their applications to biotechnology.

bioreactors including the quantity of microorganisms and

Six marine bacteria displaying QS inhibitory

the structure of the consortia as well as the increase in the

activity were isolated from 150 bacteria and were further

application of molecular techniques.

analyzed using P. aeruginosa (Busetti and Gilmore, 2010).

Using compound-specific stable isotope analysis,

The use of granules for the production of signal molecules

molecular biology, and protein based stable isotope

resulted in the attached-growth mode being dominant and

probing, Methylibium petroleiphilum PM1 was found to be

more rapid biofilm formation as opposed to when AS was

responsible for the degradation of C13 methyl-tert-butyl

used as the source of signal molecules (Ren et al., 2010).

ether (MTBE) in a mixed consortium (Bastida et al., 2010).

The sludge granulation process was also accelerated by

The bacterium was a member of the US3-M enrichment

intracellular extract from mature granules.

culture originally from a gasoline contaminated site which had been maintained on MTBE as sole carbon media.

The effect of 2(5H)-furanone and different acyl chain lengths of N-Acylhomoserine lactone (AHL) on

Breugelmans et al. (2010) showed that the linuron

biofilm inhibition was studied using Chromobacterium

degrading ability of a consortium could be decreased by the

violacium CV026 as an indicator strain (Ponnusamy et al.,

introduction of alternative carbon sources. The biofilm

2010). It was found that 2(5H)-furanone inhibited QS

composition, spatial and structural organization was

activity of the AHLs and biofilm formation.

affected by the different alternative carbon sources.

Shear. P. aeruginosa PAO1 attachment was

A lake biofilm was able to dissipate a distinct

found to have an increasing then decreasing trend with

group of Fluka naphthenic acids, but not Athabasca Oil

regards to shear in the range 1.0 to 26 mN/m2 (Raya et al.,


2010). Furthermore, rhamnolipids were found to deter

glucose starvation reduced molecular mobility, whereas the

attachment but prewashing the cells with rhamnolipids did

opposite was true for filamentous biofilms. Fragile biofilms

not have a statistically significant effect. The effects of

that were cultivated in parallel plated flow cells were

shear were also studied by Bester et al. (2010) using

stabilized for CLSM using agarose (Pittman et al., 2010).

Pseudomonas. It was observed that after perturbation which

Yawata et al. (2010) combined confocal reflection

removed

microscopy and fluorescent CLSM to visualize the effects

the shear-susceptible layer,

the non-shear

susceptible region responded with changes in nutrient and

of biofilm structure on mass transport.

oxygen flux such that CO2 production returned to previous

Surface-enhanced Raman scattering was used to

levels within 24 hours. The shear susceptible region of the

improve the sensitivity of Raman microscopy and perform

biofilm also increased CO2 production when resuspended

rapid biofilm analysis of different components in situ

without additional nutrients. A pure culture biofilm was

(Ivleva et al., 2010).

cultivated in a modified commercial biofilm reactor with

The mesoscale structures of three different

defined conditions of DO and fluid shear to show the

heterotrophic biofilms were visualized using optical

stratification of respiratory activity in a biofilm (Cotter et

coherence tomography (OCT) (Wagner et al., 2010).

al., 2010).

Volumetric porosity was found to be 0 36 for turbulent flow as compared with 0 65 for laminar and transient flow.

Techniques and Analytic Methods Imaging. Wright et al. (2010) reviewed the use of

Characteristic parameters determined using CLSM were found to differ from those obtained using OCT.

atomic force microscopy (AFM) to study the properties of

Ramanan et al (2010) showed that mass transport

surface macromolecules and the energies associated with

processes

in

biofilms

their mechanical conformation and functionality which

paramagnetic tag and

provided insight into the behavior of microbial systems and

specifically

their relationships with the environment.

diethylenetriamine pentaacetic acid (Gd-DPTA) in a

with

a

could

be

imaged

using

a

magnetic resonance imaging complex

of

gadolimium

and

Biofilm structure, composition, and molecular

phototrophic biofilm. This technique was used to show that

mobility was analyzed using confocal laser scanning

Gd-DPTA was transported purely by diffusion in agar, but

microscopy (CLSM) and C13 nuclear magnetic resonance

by diffusion and advection in the biofilm. EPS extraction techniques. Four extraction

(NMR) in order to study interactions between biofilm architecture, composition or growth states and biofilm

methods

including

ethylenediamine

tetraacetic

detachment (Garny et al., 2010). In the biomass of the non-

(EDTA), sodium hydroxide (NaOH), cationic exchange

filamentous biofilms, the molecules were found to be more

resin and ultrasound were used to extract proteins,

strongly bound than those within the detached biomass and

carbohydrates, humic substances and DNA in wastewater


acid

sludge and in biofilm which were then studied using

affecting the compositions while the chemical extraction

Fourier-transform

methods changed the protein and polysaccharide content.

infrared

(FTIR)

spectroscopy,

fluorescent in situ hybridization (FISH), and fluorescence

Using

staining (Liang, Li, et al., 2010). EDTA and ultrasound

fluorescence spectra, humic acid was found for all

were more effective extraction methods although NaOH

extraction methods while fulvic acid was only found in

and ultrasound led to the higher glucose-6-phosphate

formaldehyde with NaOH extractions.

three-dimensional

excitation-emission

matrix

(2010a)

Other techniques. A modified microcantilever

compared four methods of EPS extraction for three

method was used for measuring the strength of an intact

activated sludge (AS) samples from different membrane

biofilm (Aggarwal et al., 2010). Highly variable, but

bioreactors (MBR). They found that cationic exchange

isotropic cohesive strengths were found.

dehydrogenase

activity.

Dominguez

et

al.

resin extracted more EPS than formaldehyde, formaldehyde

GPS-X simulation software was used and showed

with NaOH, formaldehyde with NaOH and sonication, or

that the effective specific surface area for a spherical,

thermal treatment. Using three-dimensional excitation-

porous polyvinyl alcohol gel was 2500 m2/m3 and the

emission matrix fluorescence spectroscopy, protein-derived

specific surface area was 1000 m2/m3 for a comparative

compounds were found in all EPS extracted. Humic

analysis using two moving-bed biofilm reactor systems

substances were also found when formaldehyde was used.

(Levstek et al., 2010). Pulsed-field gradient NMR methods

Dominguez further studied the extracts in terms of

were used to calculate surface-averaged relative effective

molecular weight (MW) distribution and average MW with

diffusion coefficient profiles for water in live biofilms

High-Pressure Size Exclusion Chromatography and found

(Renslow et al., 2010). The coefficients decreased from the

that chemical extraction techniques did not affect MW

top to the bottom of the biofilm, differing for different aged

distribution fingerprints while physical extractions did

biofilms, generally decreasing over time, having a typical

(Dominguez et al., 2010b).

surface near the top of the biofilm, but were different for

Three-dimensional excitation emission matrix

each biofilm near the bottom.

fluorescence spectroscopy was used to show that the

A triplicated 2(3) factorial design was used to

interaction between EPS and Hg(II) was pH dependent

show that the most effective method for detaching

Zhang, Pan, et al. (2010).

culturable bacterial communities from wetland gravel

Pan et al. (2010) used centrifugation and

involved mechanical shaking for 3 hours at 30°C in a

ultrasonication as well as ethylenediaminetetraacetic acid

phosphate buffer with an enzyme cocktail (Weber et al.,

(EDTA), formaldehyde, and formaldehyde with NaOH for

2010).

EPS extraction from alga-bacterial biofilm. They found that

Spectrophotometry liquid ion chromatography

ultrasonication as a pretreatment increased yield without

and image cytometry were used to study the ammonia


oxidizing bacteria (AOB) and nitrite oxidizing bacteria

was higher for conditions of higher ratios of NO3- to

(NOB) volume and metabolic activity when immobilized in

sulphide. Acetate oxidation was found only for high ratios

an open cellular polyurethane material to create an

of NO3- to sulphide. Using FISH for the detection of

automatic biodetector for continuous monitoring of

nitrifying microorganisms in a biofilm consortia in a fixed-

nitrification (Woznica, Nowak, Beimfohr, et al., 2010). The

bed biofilm reactor, an increase in Nitrobacter species and a

method was then used to show the continuous on-line

disappearance of Nitrospira species for high nitrite (NO2-)

monitoring of the toxic effects of phenol, cyanide, a range

concentrations was found (Haseborg et al., 2010).

of heavy metals, herbicides and oxidative chain inhibitors

Alternatively, for high ammonia (NH3) concentrations, the

on

abundance of AOB increased slightly while there was no

the

nitrification

processes

(Woznica,

Nowak,

Karczewski, et al., 2010).

change in Archaeabacteria. Carriers. Wang, Liu, and Liu (2010) found that

Biofilm Reactors

the addition of biofilm carriers to a conventional AS MBR

General. A bioreactor with a Cr(III) specific

resulted in lower EPS concentrations and enhanced sludge

zeolite and the bacterium Arthrobacter viscosus was used

properties including larger particle sizes, more compact

to show that improved removal efficiency was attained

particle structures and better settleability leading to lower

when initial biomass concentration was 5 g/L in an acidic

cake layer resistance and slower transmembrane pressure

pH (Pazos et al., 2010).

(TMP) increase.

The impact of the addition of formalin on

Wolff et al. (2010) found that carbon and N

formaldehyde removal and nitrification in biofilters was

removal were affected by the carrier type and not by the

studied (Pedersen et al., 2010). Formaldehyde removal was

sludge retention time when using recycled plastic in

found to increase while nitrification was not affected in

comparison with polyethylene.

systems receiving daily treatment whereas intermittently

Woznica, Karcz, et al. (2010) used X-ray

treated systems had increased variation and reduction in

tomography, light confocal microscopy, and scanning

nitrification.

and

electron microscopy (SEM) and found that there were three

methylaminoantipyrine (MAA) with biofilms created from

distinct immobilization patterns for nitrifying bacteria on

river water was studied (Pieper et al, 2010). MAA was

polyurethane foam including large irregular aggregates,

removed rapidly while phenazone was 85% degraded, but

spherical aggregates, and threads.

The

removal

of

phenazone

only under limiting nutrient conditions in a batch reactor.

Significant colour removal was found when non-

Biofilms seeded from oil reservoir cultures were

woven material from used diapers was used as the carrier

more able to remove sulphide and NO3- than suspended

for a three stage system having approximately 60 days of

cells (Tang et al., 2010). Conversion of sulphide to sulphate

retention time for each stage (Chang et al., 2010).


Suspended solids (SS) removal was over 90% for each

exposed AS. They compared phenol mineralization

stage and the last stage had a soluble COD removal

resulting from UV photolysis alone, biodegradation alone,

efficiency of 90%. Carbon fiber felt carriers were used in a

and photolysis combined with biodegradation and found

packed reactor to remove phenol while removing N (Chen

that the latter was the most efficient for complete

et al., 2010). Nitrification was not inhibited until the

mineralization while the phenol degradation rate was equal

concentration of phenol was greater than 1200 mg/L.

for biodegradation with and without photolysis. Zhang,

Denitrification was not affected by phenol degradation.

Wang,

and

Rittmann

(2010)

performed

a

similar

Guo et al. (2010) used polyurethane foam sponge

experiment using titanium oxide with UV light for

as a carrier in three different reactors: fixed bed, batch, and

photocatalysis and again found that the most continuous

submerged MBR. The fixed bed efficiency was found to be

rapid degradation and complete mineralization was

most affected by the thickness of the sponge whereas the

achieved with the combined photobiodegradation.

batch reactor was affected by the sponge volume. The batch

A biofilm reactor with polyacrylonitrile balls as

reactor had more effect on the phosphorous (P) removal

carriers combined with an anthracite coal gravity filter was

than organic or N removal. The submerged MBR had high

used to treat a 2.4 kg COD/m3/d and was able to remove

ammonium (NH4+) and P removal.

90.6% COD, 81.4% NH3, 64.6% total N, 96.7% turbidity

Ivankovic et al. (2010) used expanded, semi-

with a 0.15kWh/m3 or 1.5 kWh/kg BOD5 power

expanded, and raw perlite as an immobilizing carrier for

consumption (Li, Yang, et al., 2010). The deposition rate

Acinetobacter junii for the removal of phosphate. While the

coefficient of C-60 nanoparticles was found to be higher

number of immobilized bacteria was highest for expanded

for porous media having a biofilm EPS coating than

perlite and lowest for non-expanded, the semi-expanded

without (Tong et al., 2010). The biochemical and physical

perlite was the best carrier in AS. Expanded perlite floated

characteristics of the biofilm EPS were found to be

at the surface, while raw perlite settled on the bottom, and

responsible for the increased C-60 deposition although

semi-expanded perlite was incorporated into the sludge.

Derjaguin-Landau-Verwey-Overbeek

When grape seeds were used as a carrier, the introduction

could not explain the enhanced deposition.

of methanol in the feed decreased the presence of fungi

Pre-treatment

with

(DLVO)

advanced

theory

oxidation.

while increasing the presence of yeast (Moreno-Terrazas et

Yapsakli et al. (2010) compared two different grades of

al., 2010). Species found included Pseudomonas, Bacillus,

granular activated carbon (GAC) and the effect of pre-

and Rhodotorula.

ozonation on the nitrification community developed in a

Zhang, Liu, et al. (2010) used a light porous

biological activated carbon packed column. They found

ceramic carrier in a photolytic circulating-bed bioreactor

that GAC grade and ozonation did not affect the

(PCBBR) to support bacteria that originated in a phenol


nitrification rate although ozonation affected the types of

for the treatment of landfill leachate achieved removal of

AOB species that developed.

approximately 85% COD, 80% N, and 70% P reaching

Fluidized Bed Biofilm Reactor. An anoxic-

effluent characteristics of less than 35 mg SBOD/L, less

aerobic twin fluidized-bed bioreactor (FBBR) was shown

than 35 mg NH4+-N/L, less than 1.0 mg phosphate-

to remove 96% organic matter, 84% N, and 12%

phosphorous (PO4-P)/L, and 37 mg VSS/L (Eldyasti et al.,

phosphorus, generating an effluent containing less than 1.0

2010).

mg ammonium-nitrogen (NH4+-N)/L, less than 4.3 mg

The addition of inorganic carbon as sodium

nitrate-nitrogen (NO3-N)/L, less than 6 mg total nitrogen

bicarbonate (NaHCO3) in an airlift-FBBR containing

(TN)/L, less than 6 mg soluble biochemical oxygen

sponge cube media resulted in the elimination of NO2-

demand (SBOD)/L, and 6-10 mg volatile suspended solids

oxidizing bacteria in the reactor and the stabilization of

(VSS)/L for organic loading rates of 1.3-2.3 kg chemical

nitritation (Tokutomi et al., 2010).

oxygen demand (COD)/m3·d, N loading rates of 0.14-0.25

A new equation for the drag force on bio-coated

kg N/m3·d, and a solids retention time of 72-108 days

particles as a function of the terminal settling velocity was

(Andalib et al., 2010b). Further study using a twin

developed based on Archimedes numbers from two

circulating FBBR had similar results compared with the

laboratory-scale liquid-solid circulating fluidized bed

previously mentioned study, but the effluent nitrification

bioreactors (Andalib, Zhu, et al., 2010). They were able to

was found to be very sensitive to soluble chemical oxygen

use the equation to accurately predict the terminal settling

demand (SCOD) concentration as the effluent NH3 level

velocity for other literature data for Reynolds’ numbers less

was increased from 1.8 to 14 mg/L after being subjected to

than 130. Computational fluid dynamics (CFD) modeling

a carbon shock (Andalib et al., 2010a). The results from a pilot scale liquid-solid

that accounted for aeration, sludge rheology, and geometry

circulating fluidized bed (LSCFB) using lava rock media

in a MBR was applied and validated against experimental

showed that approximately 90% organic, 80% N, and 70%

results. It was found that sludge settling and rheology had a

phosphorus removals were achieved at loading rates of 4.12

minimal impact on bulk mixing and thus the residence time

kg COD/m3·d, 0.26 kg N/ m3·d, and 0.052 kg P/ m3·d,

distribution (Brannock et al., 2010).

respectively without the use of chemicals (Chowdhury,

An AQUIFAS biofilm diffusion model was

Nakhla, Zhu, and Islam, 2010). In a another experiment, the

developed, calibrated, and validated for a liquid/solid

LSCFB was able to respond favourably to dynamic

FBBR

loadings by simulating a wet weather condition using a

anaerobic/aerobic

peaking factor of 3 for 4 hours (Chowdhury, Zhu, and

Nakhla, Sen, and Zhu, 2010).

using

pilot-scale reactor

results

for

arrangement

an

anoxic-

(Chowdhury,

Nakhla, 2010). Further experiments with the same reactor


Packed Bed Bioreactors. Dizge and Tansel

kg/m3·d. FISH was used to show that the constituent

(2010) were able to predict the removal of carbohydrate

bacterial members were the same as those of the indigenous

and protein in a packed bed bioreactor (PBBR) containing

inoculate

AS bacteria that were immobilized on microporous

Nitrospira. A PBBR filled with polyethylene tape as

polyurethane cubes. They determined rate constants using

packing media was studied for the treatment of domestic

external film diffusion models for various flow rates.

wastewater with and without effluent recycling (Nacheva

including

Nitrosococcus,

Nitrobacter,

and

A re-circulating packed bed batch reactor

and Chavez, 2010). The reactor obtained 90-95%

(RPBBR) using calcium alginate immobilized Bacillus sp

(biochemical oxygen demand) BOD5 removal and 69-72%

was used to study the effect of flow rates on external mass

TN removal with low sludge yield. Continuous nitrification

transfer coefficients for the bioreduction of Cr(VI) based on

of synthetic seawater in two fixed-bed reactors was

external

obtained with either polyethylene/clay sinter lamellas

film diffusion

models

as

compared

with

experimental values (Kathiravan et al., 2010).

(PBBR A) or porous ceramic rings (PBBR B) as the media

The nitrification process was investigated in a

inoculated with Nitrosomonas and Nitrospira enriched

PBBR containing either inert or carbonaceous media

from seawater and sediment samples. The NH3 oxidation

(Kowalski et al., 2010). For inert media, nitrification

rate of the PBBRs was at least 10 times higher than that for

stopped when the alkalinity of the bulk liquid was

suspended cultures. PBBR A achieved a higher NH3

exhausted, but for carbonaceous media, the dissolution of

oxidation rate and NO2- oxidation rate while PBBR B

the media allowed pH control within the biofilm.

achieved

Dispersion modeling and reactor in series modeling was

accumulation. There was a shift in the Nitrosomonas in

used to estimate nitrification rate constants from the

PBBR B for an increasing NH3 loading rate and a

reactors.

decreasing pH. The hydraulic residence time (HRT) and the

higher

The

overall

nitrification

biodegradation

of

without

benzene,

NO2-

toluene,

concentration of feed phenol were shown to determine the

ethylbenzene, and xylene compounds was modeled for a

phenol removal efficiency in a modified three-stage airlift

fixed bed biofilm reactor and compared with literature data

PBBR containing a flexible packing material (Huang et al.,

(de Mello et al., 2010). The model comprises a single

2010). The reactor was able to achieve 100% phenol and

differential equation describing the concentration profile. A

90% COD removal for a surface loading rate of up to 2.84

kinetic model was created for a pilot-scale fixed-biofilm

g phenol/m2·d.

reactor containing polyurethane foam sponge cubes (Lin,

A PBBR with indigenous nitrifying bacteria was

2010). The model described the relationship between the

used for the treatment of NH3 in recirculating aquaculture

influent substrate level, the effluent COD, the concentration

water (Kumar et al., 2010). NH3 removal averaged 0.153

of suspended biomass and the amount of CO2 of a textile


wastewater. A model was developed for a biological

Ferrai et al (2010) modeled a large pilot-scale

aerated filter that represented the physical and biological

MBBR that was used for treating pre-settled municipal

phenomena of filtration for a semi-industrial pilot-scale

wastewater. Respirometric tests were used in an extended

submerged biofilter used for tertiary nitrification for the

version of ASM3 to show that storage was the prevalent

effluent of a municipal AS plant (Vigne et al., 2010).

mechanism for heterotrophic biomass.

Moving Bed Biofilm Reactor. Kaindl (2010)

The thickness of the mass transfer boundary layer

discussed the upgrading of an AS plant with the addition of

was found to be dependent on the bulk flow rate in a

a moving bed biofilm (MBBR) reactor upstream of the AS

MBBR using a one dimensional mathematical model and a

process and ozonation and subsequent biofiltration

microelectrode measuring the oxygen profile (Masic et al.,

downstream of the AS process to address an increase in

2010).

COD load volume, peaks in COD, and colour removal in a paper mill wastewater system. The N removal rate for a MBBR used for aquaculture recycle was far lower as found in activity

Rotating Biological Contactor. A zero-order reaction rate constant of 5.7 was found for a rotating biological contactor (RBC) treating grey water for reuse as toilet flush water (Baban et al., 2010).

assays than predicted (Kessel et al., 2010). A large number

Coello et al. (2010) demonstrated the use of

of Planctomycetes bacteria were found in the biofilm that

different parameters to measure the activity of a biofilm on

did not bind with anammox bacteria probes.

a RBC including the specific oxygen uptake rate (OUR),

N removal for a landfill leachate in a MBBR was

INT-dehydrogenase activity, and the ratio of active/total

limited by the lack of biodegradable substances in the

cells. Phenol as a co-contaminant was shown to decrease

influent which could be ameliorated with the application of

the ability of a lab scale RBC biofilm to remove total

an ozone dose as high as 0.5 g/L (Plosz, Ried, et al., 2010).

petroleum hydrocarbons and total chemical oxygen demand

In a further study, ozonation installed between the pre-

(TCOD), although the phenol was completely removed

anoxic zone and the aerobic zone in a staged MBBR used

(Chavan and Mukherji, 2010). A single stage and a two

to treat a landfill leachate resulted in better COD,

stage RBC were able to complete nitrification for a loading

polyaromatic hydrocarbons (PAH), and N removal than

rate of 1.92 g N- NH4+/m2·d and 3.6 g N-NH4+/m2·d,

ozonation in the recirculation line (Plosz, Vogelsang, et al.,

respectively (Kulikowska et al., 2010). The dominant

2010).

species in the biofilm were found to be Nitrosomonas Shen et al. (2010) used a GAC airlift reactor for

europaea and N. eutropha. All four stages of a RBC used

the biological removal of ampicillin-loaded wastewater.

to treat textile industry wastewater were necessary to

They found that 60% of the removal was by adsorption

achieve textile 90.5% color and 95.3% COD removal while

whereas 40% was by biodegradation.


removal of only 75.2% of color and 85.2% of COD were

including OUR and COD removal efficiency as well as

obtained in the first two stages (Goyal et al., 2010).

molecular in situ detection methods and microscopy

A dynamic model of a rotating disk biofilm

including electron scanning microscopy and phase contrast

reactor was developed for partial nitrification which

analysis (De Sanctis et al., 2010). Mainly Proteobacteria

predicted the following: at least 40 days was required for

were found in the mature granules. The SBBGR was used

the active biomass to reach steady-state, pH affected NO2-

at a municipal wastewater plant to remove 80% of COD,

oxidizing bacteria more so than NH3 oxidizing bacteria,

TSS, and N with low sludge production (Di Ianconi et al.,

and NO2- accumulation was strongly affected by the N

2010).

loading rate and the pH (Huilinir et al., 2010).

Sequential anaerobic-aerobic granular treatment

Sequencing Batch Biofilm Reactor. Bacardit et

was used to remove 90% colour and 80% COD from

al. (2010) studied a sequencing batch biofilm reactor

coloured textile wastewater. The granules had a 3-10 mm

(SBBR) downstream of an advanced oxidation process for

diameter, a density of 66 g/L, a sludge volume index of 90

its ability to treat 4-chlorphenol. The OUR was found to be

to 130 mg/L, an integrity coefficient of 3.7, and a settling

useful for indicating the performance of the system. The

velocity of 70 m/h (Ibrahim, 2010). Granules developed in

microbial consortium was found to be varied with the

an anaerobic-aerobic SBBGR were larger, stronger, and

Wautersia genus representing about 30% of the bacteria.

had a more rapid settling velocity (Muda et al., 2010). The

Granular Reactors. Full nitritation was obtained in a continuous granular airlift reactor by maintaining a

reactor removed 94% of the COD, 95% of the NH3, but only 62% of the colour.

constant ratio between DO and total NH3-N in the bulk

The community structure of granules cultivated in

reactor fluid (Bartroli et al., 2010). Full nitritation was also

an NH3 fed aerobic upflow fluidized bed reactor started as a

obtained for high DO as long as the DO/total-ammonia-

diverse group of NH3 oxidizing bacteria that shifted to

nitrogen (TAN) concentration ratio was maintained.

predominantly N. mobilis and then to a mixed group of

Granules in a sequencing batch biofilm granular reactor

Nitrosomonas species with maturation of the granules

(SBBGR) were able to generate a stable molar ratio of NO2-

(Matsumoto et al., 2010). NO2- oxidizing bacteria were

to NH4+ of 1.0 and a maximum NO2- production of 1 g

initially represented by Nitrospira, but predominantly by

NO2-/L·D without the production of NO3- (Vazquez-Padin,

Nitrobacter after the formation of the granules. A SBBGR

Figueroa, et al., 2010). The granules had an average

was successfully used to treat saline wastewater containing

diameter of 1.9-2.9 mm.

up to 2000 mg/L phenol as the sole carbon substrate

During the transition from AS to granular sludge

(Moussavi et al., 2010). The consortium used had

in a SBBGR, changes in biomass composition were

previously been cultivated on phenol and acclimated to salt.

observed

using

traditional

metabolic

measurements


Sludge granules that were bioaugmented with P.

intermediate compound (Vasquez-Padin, Mosquera-Corral,

putida SM1443 carrying plasmid pJP4 were better able to

et al., 2010). Total solids concentrations inside the reactors

remove 2,4-D than were the controls (Quan et al., 2010).

and biofilm density were correctly described for three

Degradation followed Haldane kinetics with V-max of 31.1

different ratios of COD/N.

mg of 2,4-D/gVSS·h, K-i of 597.7 mg/L, and K-s of 257.3

Integrated

Fixed

Film

Activated

Sludge

Reactor. Biomass settleability was evaluated with two

mg/L, respectively. The exopolysaccharide material involved in the

parallel treatment trains at pilot and full scale, with and

formation of aerobic granules was shown to be a

without integrated fixed film activated sludge (IFAS)

heteropolysaccharide consisting of eight sugar residues

media (Kim et al., 2010). The attached phase density and

including alpha-galactose, alpha-rhamnose, 2-acetoamido-

polyphosphate content was lower than that of the

2-deoxy-alpha-galactopyranuronic

beta-mannose,

suspended phase. Decreases in solids retention times

beta-galactose, beta-glucuronate, beta-glucosamine and N-

resulted in the increased density and the polyphosphate

acetyl beta-galactosamine using 1D and 2D NMR and high-

content in suspended biomass but not in attached biomass.

performance anion-exchange chromatography with pulsed

Glycerol was used as the electron donor and carbon source

amperometric detection, heteronuclear multiple bond

for enhanced denitrification using a sequencing batch IFAS

correlation,

enhancement

reactor (Lu and Chandran, 2010). The glycerol kinetics was

spectroscopy (Seviour et al., 2010). The selective discharge

found to be approximately three times higher than those

of small and loose sludge flocs was found to be the key

found for methanol. The suspended phase contributed

controlling factor for aerobic sludge granulation using four

approximately 70% of the denitrification on a mass basis.

batch column reactors (Li, Zhang, and Li, 2010). Complete

C13-DNA stable isotope probing showed that the bacteria

granulation was reached with selective discharge of small

involved in assimilating the glycerol in the suspended

and slow-settling sludge flocs as opposed to unselective

phase were related to Comamonas and Diaphorobacter

discharge of mixed sludge or selective discharge of dense

whereas those in the fixed phased were related to

settled sludge. Small and slow-settling sludge flocs were

Comamonas, Bradyrhizobium, and Tessaracoccus, all of

found to compete with granulation by having better

which

substrate uptake thereby interfering with granulation if not

denitrifying bacteria.

and

nuclear

acid,

Overhauser

discharged.

were

different

from

methanol

assimilating

Tahalla et al (2010) mathematically modeled NH3

A one-dimensional biofilm model was used to

removal in a hybrid AS biofilm system using Monod

simulate a granular biomass sequencing batch reactor based

kinetics and Fickian diffusion principles. Validation of the

on the AS model platform as well as simultaneous growth

model using a laboratory scale system showed good fit with

and storage of substrates and the inclusion of nitrite as an


under prediction at high removal and over prediction for

nitritation when compared with membrane-aerated biofilm

low removal efficiency values.

reactors (Lackner et al., 2010).

Reactor Comparisons. Zhu, Getting, and Bruce

Six AS effluents were additionally treated with

(2010) reviewed biologically active filters (BAFs) used in

active carbon filtration, ozonation at two different doses,

drinking water applications including the advantages and

ozonation and MBBR and UV irradiation with hydrogen

applications of BAFs for the removal of contaminants, the

peroxide

microbial communities, and biofilm adhesion and control.

Measurements using copepods showed the low dose

(Lundstrom,

Bjorlenius,

et

al.,

2010).

An activated carbon filter was compared with a

ozonation treated effluent had the least negative effects and

ceramic granule for partial nitrification of non-rich

the addition of the MBBR did not improve the effect. In

wastewater (Wang, He, and Yang, 2010). Effluent from the

another study, Lundstrom, Adolfsson-Erici, et al. (2010)

ceramic granule reactor had a higher NO2-N and was more

found that the MBBR resulted in the lowest negative

resistant to fluctuations in influent COD/N ratios. An AS

effects. Ozonation was the most efficient treatment system

system was compared with a hybrid moving bed biofilm

for pharmaceuticals although it caused negative effects in

reactor

the ecotoxicological tests.

(HMBBR)

using

comparable

mixed

liquor

suspended solids (MLSS) (Di Trapani, et al. 2010a).

Effluent from two FBBRs and two MBBRs from

Although the HMBBR had twice the hydraulic load as the

a rainbow trout facility was compared. When expressed

AS system, the HMBBR was found to perform similarly to

based on reactor volume, MBBR filters had a greater total

the AS system in terms of organic and N removal. A pilot

TAN removal than the FBBRs, but not when expressed

plant having two parallel lines was operated in Acqua dei

based on surface area (Suhr and Pedersen, 2010). FBBR

Corsari (Polermo) comparing the AS process and HMBBR

reactors were also able to increase removal with increased

using a 30% fill ratio for the purpose of determining kinetic

TAN pre-conditioning. The variation in the particle size

heterotrophic constants using respirometry (Di Trapani et

distribution associated with the secondary biofilm treatment

al., 2010b).

systems at three different urban wastewater treatment

A submerged biofilter system, trickling filter

plants was found to be related to the COD, SS, and the

system, and a rotating biological contactor system were

mean particle size (Garcia-Mesa, Poyatos, Delgado, and

compared on the basis of the particle size distribution

Hontoria, 2010). The greatest reduction in particle number

(Garcia-Mesa, Poyatos, Delgado-Ramos, Munio, et al.,

was realized in the following order: submerged biofilter,

2010). Correlations between the particle size distribution

trickling filter, and a rotating biological contactor.

and SS, turbidity, and COD were determined. Conventional

Modeling. Wang and Zhang (2010) reviewed 30

biofilm reactors were found to have superior geometry for

years of literature on mathematical modeling of biofilms. They used four classes of models including low-


limited

and continuous bioreactor respectively using P. putida and

aggregation models, continuum-discrete models, and fully

calcium alginate beads (Mathur et al., 2010). A low

coupled biofilm-fluid models as well as hybrids thereof.

complexity mathematical model was generated based on an

Boltz, Morgenroth, and Sen (2010) provided a consensual

N-tanks-in-series hydraulic model and biodegradation

report of the current state of modeling including the areas

kinetics from respirometry experiments for the purpose of

of uncertainty and requirements for advancements in the

modeling compact individual wastewater treatment systems

use of modeling for design of biofilm reactors. They

(Moelants et al., 2010). Orgogozo et al. (2010) found that

considered simulators which have been augmented with a

the

submerged completely mixed biofilm reactor module based

consumption model was consistent with direct numerical

on a one-dimensional biofilm.

simulation for high Peclet and Damkohler numbers

dimensional

continuum

models,

diffusion

validity

domain

of

the

reaction-rate

limited

Boltz and Daigger (2010) discussed the two

whereas the mass transfer limited consumption model was

fundamental concepts of reactor design: bioreactor

limited to high Damkohler and low Peclet numbers. NO2-

hydrodynamics and biofilm dynamics. They stated that

accumulation was modeled for a multi-species nitrifying

hydrodynamics affects thickness control, surface area, and

biofilm in a completely mixed biofilm reactor (Park et al.,

development whereas biofilm dynamics affects thickness,

2010). The suppression of NO2- oxidizing bacteria was

structure and function. Mechanistic models were possible

found to be controlled by the total NH3-N concentration,

for IFAS and MBBR technology based on these concepts,

the bulk DO concentration, and the buffer concentration,

but not for trickling filters and biological reactors. With

while the bulk liquid pH had a weaker influence.

regards to dynamics, fixed films were found to be diffusion

The adsorption of phenol on an Arthrobacter

limited while suspended growth systems were kinetically

viscosus biofilm supported on GAC was found to follow

limited. A new annotation system was proposed to simplify

the Sips model while the one for chlorophenol fit the

documentation,

Freundlich model (Quintelas et al., 2010).

implementation,

and

connection

of

different models (Corominas et al., 2010). A one-equation

Fouling of Membrane Biological Reactors. Xia

non-equilibrium model was developed for describing the

et al. (2010) found that membrane fouling occurred faster

transport of a solute in a porous media (Davit et al., 2010).

under higher organic loading conditions although the

The model does not require the local mass equilibrium

similarity between the bacterial communities was higher

approximation but instead used an assumption that

between the membranes than between the mixed liquors.

anomalous behaviors of third and higher spatial moments

Beta-Proteobacteria

can be neglected at sufficiently long times. The parameters

predominant microorganisms in the high loading biofilm.

and

Bacteroidetes

were

the

for a Haldane model and a modified Haldane model were

Ng, Lin, et al. (2010) studied the role of SS, EPS,

generated to describe p-cresol inhibition in a batch reactor

and soluble microbial products (SMP) in membrane fouling


in a fixed carrier biological system (FCBS) and an AS

polysaccharide, proteins and DNA, energy uncoupling, use

process. They found that SMP controlled fouling while SS

of cell wall hydrolases, and bacteriophage disruption of

and EPS had a negligible effect. FCBS had a higher

biofilms.

permeate flux than AS related to the lower SMP level in

Vrouwenwelder, Kruithof, and Van Loosdrecht

FCBS. Molecular weight fractions less than 10 kDa were

(2010) proposed an integrated approach for controlling

suggested as the possible source of the fouling. Ng

and

biofouling based on equipment design and operation,

Ng (2010) showed that under a low flux condition,

biomass growth conditions, and cleaning agents. They

membrane fouling occurred through a biofilm related

suggested reducing cleaning and inhibitor dosing through

process, whereas under higher flux conditions, organic

the use of biofouling-tolerant, spiral-wound membrane

fouling became more important. The protein fraction on the

systems, capillary membranes, and the limitation of

membrane surface had the greatest impact on the increasing

phosphate.

transmembrane pressure in the early stages of membrane operations.

Zhu, Bai, et al. (2010) found that immobilized and reduced Ag on the surface of a chitosan membrane was

Juang et al. (2010) found Arthrobacter sp.

more stable than non-reduced Ag. The Ag treated chitosan

generating a biofilm inside the hollow-fiber membrane of a

showed greater anti-bacterial performance than chitosan

MBR. They suggested that the Arthrobacter biofilm in an

alone, with the reduced form showing a better long term

aerobic granular reactor was responsible for 58.9 and

performance. Wang, Yang, and Zhang (2010) created a

81.3% of the total membrane resistance when operated in a

non-woven composite membrane with chitosan coating,

batch mode or a continuous mode, respectively.

glutaraldehyde crosslinking, and pre-treatment with ozone.

Cleaning of Membranes. Poyatos et al. (2010)

Using FTIR/ATR, XPS, water contact angle, and

studied particle numbers in the permeate stream of a bench-

environmental SEM, they determined that hydrophilicity of

scale MBR under various levels of MLSS with and without

the surface was improved.

chemical cleaning. They found that the number of particles

Yang et al. (2010) found that initial bacterial

in the permeate increased as backwashing was performed

adhesion can be completely suppressed on microporous

without chemicals, but decreased after cleaning with

polypropylene membranes that have been hydrophilized by

chemical products.

UV-induced grafting of polysulfobetaine methacrylate to a

Antifouling technologies and modifications.

density of 560 µ/cm2 using a combination of conventional

Xiong and Liu (2010a) reviewed the reduction of

benzophenone entrapment and covalent immobilization.

biofouling

of

They used FTIR/attenuated total reflectance (ATR), X-ray

microbial attachment including QS, nitric oxide-induced

photoelectron spectroscopy (XPS), field emission scanning

through

biological

control

strategies

biofilm dispersal, enzymatic disruption of extracellular


electron microscopy, water contact angle, and pure water

through

flux measurements to characterize the surface.

biocathodes that were efficient for use in full-sediment

inoculation

with

active

microorganisms,

Membrane modeling. Mathematical modeling

MFCs could be developed within days. Carbon felt

with Monte Carlo filtering was utilized for the evaluation

biocathodes were found to be the best when used for fresh

of MBR operating conditions that would result in NO2-

water, while the use of stainless steel became more

oxidizers

advantageous at higher salinity applications.

being

out-competed

and

the

long-term

maintenance of partial nitrification. The main mechanism

The dependence of the electrode material and

for displacing NO2- oxidizers was competition for oxygen

temperature on the maximum achievable current density

not the difference in maximum growth rate. NH3 inhibition

from mature wastewater-based microbial biofilms was

caused faster washout but was not necessary to reach

studied by Liu, Harnisch, et al. (2010) with results

washout (Brockmann and Morgenroth, 2010). Cellular-

indicating that electrodes with large microbially-accessible

automata and individual-based models were compared for

surfaces, like carbon fiber veil or carbon-paper based

membrane processes for their ability to simulate the growth

materials, achieved 40% higher current density than on

and development of biofilm structure for different

graphite rods, but bacteria could not form well on graphite

environmental conditions (Laspidou et al., 2010).

foil. Also, raising the temperature from 30 to 40 °C resulted

A 3-D CFD model was used to predict that the

in an 80% increase in current density on graphite rod

dominant biomass growth was on the feed spacer as

anodes. Mao et al. (2010) found that a MFC could

previously observed with magnetic resonance imaging

successfully run with a biocathode, biocatalyzed by

(Vrouwenvelder, Picioreanu, et al., 2010). Feed spacer

ferro/manganese-oxidizing

fouling was shown to be more important than membrane

conditions with a maximum open circuit voltage of 700 to

fouling.

800 mV, maximum power density of 32 W/m3 and 28

bacteria,

under

batch

fed

W/m3 for batch-fed and continuous systems, respectively. Microbial Fuel Cells Fuel

Cell

Improved Electrodes.

procedures

of

inoculation

and

acidophile

conditioning of a MFC with S. oneidensis MR-1 and a

microorganism Acidithiobacillus ferrooxidans was used as

sodium lactate electron donor were presented by Prakash et

a biocathode by Carbajosa et al. (2010) and was grown

al. (2010). Single cell cultured MFC showed the good

without the addition of redox mediators, generating up to 5

performance with a typical power density of 300 mW/ m2

A/m2 current densities for O2 reduction at the cathode at pH

and a maximum value of 600 mW/ m2. Sanchez et al.

2.0. As compared with uncoated graphite electrolytes, the

(2010) a platinum nanoparticle infused single wall carbon

electrocatalytic wave was shifted to a higher potential by

nanotube sheet electrode for use as a MFC cathode. Their

300 mV. De Schamphelaire et al. (2010) found that

results showed an order of magnitude improvement in the

The


current density with the new electrode compared to an

producing mediator in a cubic air-chamber MFC was

electron beam evaporated platinum cathode, likely due to

studied using K. pneumonia (L17) as the biocatalyst (Deng

the increased available catalytic surface area in the former.

et al., 2010). Electricity generation was suggested to be due

The performance of an MFC was found to improve by a

to an electron-shuttle mechanism rather than a biofilm

lower interfacial charge transfer resistance and production

mechanism in the membrane coated MFC, with cyclic

of a higher power density when using a layer-by-layer

voltammetry measurements showing the presence of an

assembled multi-wall carbon nanotube with polyelectrolyte

electrochemically active compound produced by the

polyethyleneimine

bacteria, and identified by gas chromatography mass

modified

carbon

paper

electrode,

compared to a bare carbon paper electrode (Sun, Zhao, et

spectrometry

al., 2010).

benzoquinon. Polarized electrodes were placed in open

(GC-MS)

to

be

2,6-di-tert-butyl-p-

Sun, Zhang, et al. (2010) improved the poor

seawater so that biofilms of natural bacteria that catalyzed

biocompatibility of gold, a highly conductive and stable

the electrochemical reduction of oxygen could be formed

material, when used as an anode by homogeneously sputter

and subsequently used as the inoculum in closed

coating gold onto a carbon paper matrix. The results

electrochemical reactors, thereby allowing, for the first

indicated that both improved electric charge production,

time, marine biofilms that are able to catalyze oxygen

and accelerated S. oneidensis MR-1 biofilm growth. Zou et

reduction to be formed in small, closed vessels, although

al. (2010) found that using nanostructured, electrically

low nutrient availability hindered the reactor (Erable,

conducting fibrillar polypyrrole as an anode material

Vandecandelaere, et al., 2010). Using closed laboratory

exhibited

systems, Faimali et al. (2010) attempted to verify the

better

performance

than

using

granular

polypyrrole for electron collection from photosynthetic

relationship

biofilm in photosynthetic MFCs.

electrochemical activity by examining the diverse microbial

Characterization of Fuel Cell Biofilm. By

between

biofilm

composition

and

populations with denaturing gradient gel electrophoresis

utilizing a transitory electrochemical technique (cyclic

(DGGE).

voltammetry), Cournet et al. (2010) studied the microbial

electrochemically active biofilms with very high genetic

catalysis of the electrochemical reduction of oxygen and

variability and in some cases showed performances similar

demonstrated that a wide range of both Gram-positive and

to open systems. Franks and Nevin (2010) discussed the

Gram-negative aerobic and facultative anaerobic bacteria

mechanism of a matrix of bacterially produced nanowires

were able to catalyze oxygen reduction with direct transfer

and/or

of electrons.

sulfurreducens was thought to allow for long distance

By coating the anode electrode with a MF

The

c-type

closed

systems

cytochromes,

by

all

yielded

which

marine

Geobacter

electron transfer, and additionally discussed genetic

membrane, the mechanism of electron shuttle via a self-


engineering of this bacterium and its successes in

surface polysaccharide biosynthesis gene cluster that

increasing power output.

encodes for a putative formyltransferase. TEM images

The metabolic activity of biofilm cells close to

showed an impairment of the layered structure at the cell

and far from the surface of an anode were evaluated by a

surface that allowed the mutants to attach to graphite felt

microtoming/microarray

anodes better than wild cells, and show increased

strategy

and

found,

by

transcriptional analysis, that most genes were more active

hydrophobicity.

closer to the anode, but cells throughout the biofilm were

High throughput FLX Titanium pyrosequencing

still metabolically active, and could contribute to current

was used by Lee et al. (2010) to determine the commonly

production (Franks et al., 2010).

existing microbial populations in MFC anode biofilms

Through using inoculant designed to enrich either

subjected to different substrate exposure and respiration

aerobes/facultative species or anaerobes, Ieropoulos et al.

conditions. Through analysis of 31,901 sequence reads with

(2010) examined the effects of flow rates on the

an average length of 430 base pairs obtained from the 16S

development of anodic biofilm. The performance of all

rRNA gene, Firmicutes and the alpha, beta, gamma, and

MFCs was hindered at high flows which SEM imaging

delta subclasses of Proteobacteria were found to be the

revealed was due to clumpy growth, however, the aerobic

most common bacteria.

biofilms improved after initial development, and an

Using sodium acetate as substrate, a mediatorless

overshoot in the power curves was observed and attributed

MFC, constructed with Geobacter metallireducens, was

to an increase in internal resistance due to ionic depletion

studied for factors affecting biofilm structure and formation

and/or microbial exhaustion.

by Li, Feng, et al (2010). Results showed that current was

Using the 16SrRNA gene and FISH, the

generated by the chemically (not electrostatically bound)

microbial population of a MFC that was fed with formic

anodic bacteria, with shear stress affecting the biofilm

acid for one year was analyzed (Kiely et al., 2010). The

structure. The substrate concentration was found to be

Paracoccus genus was found to comprise approximately

proportional to electricity generation and the MFC could

30% of the anode bacterial community, although the

handle temporary intermittence.

presence of small amounts of other bacteria were believed

Li, Zhang, et al. (2010) found that both a two-

to be partially responsible for the increased power

chamber MFC and a single-chamber air-cathode MFC

production of the biofilm, as compared to cultured cells of

could remove various mixes of veratryl alcohol combined

a Paracoccus isolate. Kouzuma et al. (2010) showed that

with glucose, along with between 75% and 88% of the

strain 4A, a S. oneidensis MR-1 mutant grown in a MFC

COD. DGGE profiles of the 16SrRNA gene showed non-

fed by lactate, exhibited rough morphology, and also

dramatic changes in the dominant species of bacteria

increased current due to the SO3177 gene located in a cell

between the different substrate mixes, but the abundance of


some species did greatly increase, while SEM images

Enterococcus faecium with either of two different Gram-

showed that bacilli were the most abundant bacteria with

negative organisms, increased by 30-70% relative to strictly

the Bacteroidetes and Proteobacterium dominating.

single cultures. Results also indicated that co-culture

The physiology of the extracellular electron

biofilms segregated in time, evidenced by P. aeruginosa

transfer of G. sulfurreducens was examined during many

creating towers through a thin, uniform layer of

stages of growth for its response to changes in electron

Enterococcus faecium.

acceptor potential (Marsili et al., 2010). Results showed

A biofilm bacterial population, taken from

that the electron accepting biofilm had consistent

stainless steel electrodes that were polarized at -200 mV

electrochemistry, growth rate, and growth yield at all

versus a Ag/AgCl reference electrode and placed in natural

growth stages, indicating that there was an initial phase of

seawater, with a resulting steady current of 0.5 A/m(2),

surface attachment optimization, but afterwards, the rate of

were examined by whole cell fatty acid methyl ester

electron production by cells was governed by the rate that

analysis followed by 16S rRNA gene sequencing and

electrons could be transferred between cells, and to the

DGGE (Vandecandelaere et al., 2010). The analyses

electrodes.

revealed the presence of microbes from alpha and gamma

An optically accessible, dual anode, continuous

Proteobacteria,

the phylum Firmicutes,

the family

flow MFC that enabled microscopic imaging of anodic

Flavobacteriacae and the phylum Actinobacteriam, and

biofilm populations throughout their development from

that the dominant microbes found in natural seawater were

single attached cells to mature biofilms, was presented by

the same ones dominant in the cathodic biofilm.

McLean et al. (2010), and was subsequently used to

Phylogenic analysis and a dilution to extinction

characterize how external resistance differences affected

technique were used to analyze microbial communities in

electron transfer rates on a per cell basis as well as biofilm

single-chamber air-cathode MFCs with various anode

development in S. oneidensis MR-1. Their results showed

surfaces using DGGE based on the 16S rRNA gene (Xing

that low external resistance yielded a higher per cell

et al., 2010). Their results indicated the dominant bacteria

electron transfer rate and thinner biofilm development, as

were

compared to a MFC run at higher resistance.

aminobutyricumthe. In a study by Xu, Duan, and Hou

Read et al. (2010) performed pure and co-culture experiments

with

Gram-positive

and

Comamonas

denitrificans

and

Clostridium

(2010), where marine biofilm formation on carbon

Gram-negative

electrodes was examined, cytochrome species were found

bacteria in order to understand how biofilms are formed in

to potentially play a key role in electron transfer. Marine

MFCs and how electron transfer is affected by their

biofilms were suggested to be able to transfer electrons to

development, structure and viability. Their results showed

electrodes under both aerobic and anaerobic conditions.

that the power output of co-cultured Gram-positive


Fuels. The ability of food industry wastes

possible to reduce the common MFC problems of poor

consisting of fermented apple juice, wine lees and yogurt

cathodic oxygen reduction and build-up of a pH gradient by

waste, combined with inoculums from anaerobic sludge

repeatedly inverting the polarity of the cell, thereby

and garden compost were evaluated for their suitability as

allowing both the anodic and cathodic reactions to occur in

fuels for MFCs (Cercado-Quezada et al., 2010). Yogurt

the same half-cell by supplying acetate and DO alternately

wastes were found to be the most suitable candidate.

to the biofilm. The anodophilic biofilm was able to cope

Mohanakrishna et al. (2010) investigated the use of acid-

with the oxygen and demonstrating the concept of an

rich effluents, taken from an acidogenic SBBR that

electrochemically active biofilm catalyzing both substrate

produced H2 by fermenting vegetable waste, as a substrate

oxidation and cathodic oxygen reduction in a single system.

for a single chamber MFC with an air cathode and non-

Through varying the influent carbon source,

catalyzed electrodes. The MFC produced 111.76 mW/m2 of

alkalinity and total dissolved solids (TDS) concentration of

power and removed 80% COD, 79% volatile fatty acids,

synthetic wastewater, and inoculating with pre-heated

78% carbohydrates and 65% turbidity.

septic tank sludge, the performance of a mediator-less and

Mode of Fuel Cell Operation. Borole et al.

membrane-less microbial fuel cell was evaluated by

(2010) demonstrated that MFCs with enriched anodic

Ghangrekar et al. (2010). Results showed that a COD

microbes have the potential for stable long term energy

removal of 85% was achievable under certain conditions.

production by measuring the changes in the anode, cathode,

An up-flow single-chambered MFC reactor with three

and solution/membrane impedances of a MFC. Results

zones consisting of a lower anaerobic digestion zone, a

showed that the anode impedance was 28 times less than

central MFC zone, and an upper effluent clarifier zone was

previously

environmental

used to produce electricity from brewery beer wastewater

stressors such as oxygen, low pH, low temperature,

with sewage micro biota and anaerobic sludge (Katuri and

inhibitor (2-bromoethanesulfonate), and changes in external

Scott, 2010). The results showed that three redox peaks at

resistance, Chae et al. (2010) examined the activity of

the cathode, under continuous polarization with a constant

MFCs, and found that temperature and pH changes did not

external resistance (0.01 k Ω), a stable current density of

effect methanogenesis. Results also indicated that although

2,270 mA/m2 was generated, cell polarization gave a peak

adding 0.1-0.27 mM of inhibitor increased the efficiency

power density of 330 mW/m2 at a current density of 1,680

from 35% to 70% and lowering the resistance reduced the

mA/m2, and the overall internal resistance of the reactor

methanogenic electron loss, oxygen stress was deemed to

was 8.0 Ω, as shown by electrochemical impedance

be the most cost effective solution for both successfully

analysis. The effect of temperature on soluble organic

inhibiting the methanogens and only slightly suppressing

matter removal and energy generation was tested on both

the exoelectrogens. Cheng et al. (2010) found that it was

single and double chambered MFCs using brewery

reported.

By

introducing


wastewater diluted in domestic wastewater (Larrosa-

The dependence and limits of temperature on

Guerrero et al., 2010). The results showed that increasing

wastewater

the temperature from 4 to 25°C increased both the COD

examined at lower and upper temperature limits of 0 and

removal (58% to 94%) and electricity generation (15.1

50°C (Patil et al., 2010). Biofilms formed at higher

mW/m3 to 174.0 mW/m3) for single chamber MFCs with

temperatures formed more quickly and exhibited higher

carbon cloth-based cathodes, while a membrane-based

electrochemical activity at higher temperatures than those

cathode configuration gave promising results for operation

incubated at lower temperatures and operated at higher

at 4°C, with a maximum power output of 294.6 mW/m3.

temperatures. In order to reduce the negative effects of low

derived

anodic

microbial

biofilms

was

Carbon anodes made of graphite, sponge, paper,

pH feedstock, Sleutels et al. (2010) devised an extra

cloth, felt, fiber, foam, and reticulated vitreous carbon

compartment for bioelectrochemical systems that rests

(RVC) were tested for their effects on the performance of

between the anode and cathode, separated by cation and

MFCs fed with brewery wastewater diluted with domestic

anion exchange membranes, respectively, allowing for the

wastewater (Larrosa-Guerrero et al., 2010). In an open

clean hydrogen production. Srikanth et al. (2010) found

circuit the anode materials appeared to have an influence

that the performance of a MFC was improved by increasing

on the biofilm diversity. However, in closed circuits the

the applied potential during startup by up to 600 mV and

current passage was the determining factor of diversity.

then decreasing thereafter, due to the enrichment of the

Li, Sharma, et al. (2010) studied the effects of

electrochemically active consortium on the anode.

configuration, electrolyte solutions, and electrode materials

Using different cathode potentials at startup in

on the performance of electrogenic bacteria in terms of

order to measure MFC performance and catalytic behavior

voltage generation, power density, and acclimation period.

via polarization curves and cyclic voltammetry, Ter Heijne

Their results indicated that as compared to conventional

and Strik (2010) found that biocathodes held at +0.05 V

two chamber configurations, membrane-less two-chamber

and +0.15 V versus Ag/AgCl almost immediately produced

MFCs showed lower internal resistance, shorter bacterial

current after inoculation, but a biocathode held at +0.25 V

acclimation periods, and a higher voltage generation.

versus Ag/AgCl produced no current until day 15, with

Changing the external resistance of a MFC had the effect of

cyclic voltammetry showing clear catalysis for all three

changing the anodic biofilm community, but to a different

biocathodes. The authors noted that the highest current

composition than what was seen if the MFC had operated

density was achieved for the +0.15 V cathode potential, and

consistently at that external resistance, while biofilms

that oxygen mass and charge transfer were the limiting

established under different external resistances, resulted in

factors for the biocathodes. Wang, Sun, et al. (2010)

similar power production (Lyon et al., 2010).

showed that a MFC inoculated with an enriched anodophilic consortium performed better (higher power


densities and Coulombic efficiencies and shorter startup

recycling of H-2 between the cathode and the anode from

time) than a MFC inoculated with cells from its original

reoxidation of H2 by anode-respiring bacteria.

biofilm, or one formed from AS.

A biofilm community analysis of an ethanol fed

Microbial Electrolysis Cells. Ajayi et al. (2010)

MEC, operated with and without methanogenesis (absence

found that the effects of increasing the Reynolds number

or presence of methanogen inhibitor 2-bromoethane

from 900 to 4900 in a microbial electrolysis cell (MEC)

sulfonate) was performed (Parameswaran et al., 2010).

increased the production of hydrogen (H2) by about 30%.

Results showed that with methanogenesis, H2-oxidizing

They also found that prolonged exposure of the anodic

methanogens were the H2 scavengers, but without

bacterial communities to oxygen did not rid the culture of

methanogenesis, homoacetogens (present only without

methanogenic

(2010)

methanogenesis) were a channel for electron flow from H2

demonstrated a proof of concept whereby the anodic and

to current through acetate. Wang, Liu, Ren, et al. (2010)

cathodic reactions of a MEC were catalyzed with

measured the impact of various parameters including pH,

microorganisms, without any chemical catalysts. Their

substrate and applied voltage on the microbial anode

results indicated that H2 production was catalyzed at the

potential in order to optimize H2 production in an “H”

biocathode by electrochemically active microorganisms.

shaped MEC. Results showed that the lowest anode

bacteria.

Electrode

and

Jeremiasse

ohmic

et

energy

al.

losses

were

potential was achieved at a pH of 6.8-7.0. A pH less than

characterized by Lee and Rittmann (2010a) in an upflow,

6.0, or a substrate concentration less than 50 mg/L led to

single-chamber MEC that produced 0.57 m3-H2/ m3·d at an

decreased H2 production.

applied voltage of 1V, a cathodic conversion efficiency of

Fuel Cell Reviews. Bretschger et al. (2010)

98% and a H2 yield of 2.4 mol H2/mol acetate, without a

discussed how knowledge gained in MFC research can be

cathodic metal catalyst. Removing the membrane led to an

applied to in the context of methanogenesis and microbial

ohmic energy loss of 0.005 V, a pH energy loss as small as

fermentation in ruminant animals that are highly dependent

0.072 V, a significant cathode energy loss of 0.56 V (due to

on the consumption and production of H2 in the rumen by

the lack of catalyst), resulting in an anode energy loss of

microbes. Erable, Duteanu, et al. (2010) discussed how

0.395 V, and a 75% energy conversion efficiency. Lee and

electro-active biofilm that can be found naturally in soils

Rittmann (2010b) tracked the expected and harvested H2

and sediments, as well as other microbial rich environments

production in a single-chamber MEC that used a high

like sewage sludge, AS, or industrial and domestic effluents

density of carbon fiber catalysts as the anode. Volumetric

have recently been used for bioremediation, biosynthesis

current densities of 1470 +/- 60 to 1630 +/- 50 A/ m3 with a

processes, biosensor design, and biohydrogen production.

hydraulic retention time of 1.6-6.5 h were obtained.

Research on MFCs was reviewed by Franks and Nevin

Captured H2 was substantially lower than predicted due to

(2010), with discussions focusing on the limited practical


use of marine sediment MFCs, the push for increased

account the populations of methanogenic and anodophilic

understanding of their limitations and microbiology, and

bacteria was developed with ordinary differential equations

the degradation of toxic wastes and chemicals. A review

to describe their growth and retention, with parameters

paper on the advantages of MFCs run by microbial

chosen from and validated by four continuous-flow air-

electricigenic respiration was presented by Liu, Shao, et al.

cathode MFCs operated at several organic loads and

(2010). The principles of MFCs, species of electricigens,

external resistances (Pinto et al., 2010).

factors affecting electricigen respiration, a comparison between Fe(II) and electricigen respiration and intra and

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