Baj Pai 1997

Reduction of Organochlorine Compounds
in Bleach Plant Effluents
Pratima Bajpai and Pramod K. Bajpai

Chemical Engineering Division, Thapar Corporate Research and
Development Centre, Patiala - 147 001, India
List of Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216
2 Environmental Impact of Organochlorines . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
3 Reducing the Generation of Organochlorine C o m p o u n d s . . . . . . . . . . . . . . . . . . 226
3.1 Enzymatic Prebleaching . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
3.1.1 Hemicellulase Enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
3.1.2 Ligninolytic Enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
3.2 Fungal Prebleaching . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230
4 Treatment of Bleach Plant Effluents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235
4,1 Biological Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236
4.1.1 Using Bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236
4.1.1.1 Aerobic Processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236
4.1.1.2 Anaerobic Processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237
4.1.2 Using Fungi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238
4.2 Enzymatic Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247
5 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251
6 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251
Organochlorines have been a matter of concern in the pulp and paper industry for the last two
decades. These c o m p o u n d s are produced mainly by the reactions between residual lignin present in
wood fibres and the chlorine used for bleaching. Some of the organochlorine c o m p o u n d s are found
to be toxic, mutagenic, persistent, bioaccumulating and cause h a r m in biological systems.
Earlier measures taken by the pulp a n d pape r industry to solve the chlorine problem have
focussed on improving effluent treatment methods. Today, the emphasis of research and develop-
ment work in this area has shifted more towards improving the processes. In the search to produce
pulp with non-polluting chemicals, more efficient pulping m e t h o d s reducing the a m o u n t of residual
lignin passing to the bleaching process and alternative bleaching methods, are being developed.
There are also possibilities for treating effluent with microorganisms and enzymes to remove the
dechlorinated organic material. Each option has inherent advantages and disadvantages with regard
to capital costs, operating costs, ease of retrofit, fabrication a n d installation time. Impact on other
mill unit operations is also considered in choosing the best options. M a n y factors have to be
considered in choosing an effective and economical bleaching/treatment process that meets all the
environmental guidelines.
This review describes the environmental impact of organochlorines, use of enzymatic and
biological bleaching for reducing the generation of organochlorine compounds, and treatment of
bleach plant effluents by biological and enzymatic methods. Advantages and limitations of various
biotechnological methods are discussed.
Advancesin BiochemicalEngineering/
Biotechnology,Vol. 57
ManagingEditor: T. Scheper
9 Springer-VerlagBerlinHeidelberg 1997
214 P. Bajpai and P.K. Bajpai
List of Abbreviations
ADI Acceptable daily intake

AOX Adsorbable organic halogens
BOD Biological oxygen demand
COD Chemical oxygen demand
CTMP Chemi-thermomechanical pulp
DDT Dichlorodiphenyltrichloroethane
ECF Elemental chlorine free
EOC1 Extractable organic chlorine
EOX Extractable organic halogen
EPA Environmental protection agency
HC High concentration (consistency)
HRT Hydraulic residence time
ISO International standards organization
LMS Laccase mediator substrate
MBC Modified batch cooking
MCC Modified continuous cooking
MW Molecular weight
OKP Oxygen bleached kraft pulp
PCB Polychlorinated biphenyl
PCDD Polychlorinated dibenzo dioxins
PCP Polychlorophenol
Pow Octanol/water partition coefficient
ppb Parts per billion (1/109)
ppm Parts per million (1/106)
ppt Parts per trillion (1/1012)
RBC Rotating biological contactor
t metric ton
TCDD Tetrachlorinated dibenzodioxins
TCF Totally chlorine free
TCP Trichlorophenol
TEQ Toxicity equivalent
TMP Thermomechanical pulp
TNT 2,4,6-Trinitrotoluene
TOC1 Total organochlorine
Bleachin9 stages
C Chlorination
(CD) Treatment by mixing chlorine and chlorine dioxide simultaneously
(C1 proportion is higher than D)
D Chlorine dioxide treatment
D1 & D2 First and second treatment, respectively with chlorine dioxide
Reductionof OrganochlorineCompoundsin Bleach Plant Effluents 215
(DC) Treatment by mixing chlorine dioxide and chlorine simultaneously

(D proportion is higher than C1)
E Alkaline extraction
E1 & E2 First and second alkaline extraction respectively
Eo Alkaline extraction in presence of oxygen
Eov Alkaline extraction in presence of oxygen and hydrogen peroxide
Ev Alkaline extraction in presence of hydrogen peroxide
F Fungal treatment
H Hypochlorite treatment
L Laccase enzyme treatment
O Oxygen delignification/bleaching
P Hydrogen peroxide treatment
Q Chelation of metal
X Xylanase enzyme treatment
Z Ozone treatment
1 Introduction
The bleaching of pulp became an issue of great concern during the 1980s
primarily because of growing alarm over chlorinated organic compounds,
referred to as adsorbable organic halogen (AOX), in bleach plant effluents.
In particular, the detection of chlorinated dioxins and furans led to strong
reactions.
The paper industry recognized chlorine bleaching to be a potential problem.
As the analytical instruments and methods became available, especially for
chlorinated organic substances, studies were made on the chemical composition
of bleach plant effluents. The new analytical methods and studies also created
a basis for governmental regulations. The focus has been on setting upper limits
for AOX/TOC1. At present, the strictest general emission limit value for AOX is
1 kg/t of pulp. This limit applies to sulphite pulp e.g. in Austria, Germany and
Norway. This is because AOX emissions are easier to reduce with sulphite pulp
than with softwood kraft pulp. However, in Sweden, kraft pulp mills have
individual limits as low as 0.3 kg/t pulp. Some decided or planned regulations
for AOX are shown in Table 1 [1]. The permitted levels are likely to decrease to
about 1 kg/t in the next few years in many countries. In several countries or
provinces, emission limits are decided mill by mill taking into account the local
Table 1. Regulations for discharge of chlorinated organic compounds

measured as AOX (kg/t pulp) from bleaching of chemical pulps
Country 1994 1995-2000 2000-2005
Australia 1.0a
Austria 0.75 1.5 0.5 1.0
Belgium 1.5
Canada
- Alberta 0.29a/1.5
British Columbia 1.5 0
Ontario 1.5 0.8
Quebecb 1.5 (HW) 1.0-2.0
Finlandb 1.0-2.0
Germanyb 1.0
India~ 2.0
Norwayb 1.0-2.0
Japanc 1.5
USAa 0.156
Sweden 1.2-1.5 0.3-1.0 0.3-0.5
"Limits for new mills
bLower limits for hardwood pulps
cGuidelines of Japan Pulp, Paper and Paperboard Association
a Proposed regulation on "Cluster Rules" by EPA
eProposed by Ministry of Environment and Forest, Government of India
Based on data from Ref. [1]
Reduction of OrganochlorineCompoundsin Bleach Plant Effluents 217
factors. Thus, for individual mills stricter emission limits may apply than those
given in the table. According to a decision in 1992 by PARCOM (Paris
Convention for Prevention of Marine Pollution from Land Based Sources and
Rivers) signed by twelve European countries, a general AOX emission limit
would be 1 kg/t from 1995. The limit applies for all types of bleached chemical
pulp and has been accepted by Belgium, Denmark, France, Germany, Great
Britain, Ireland, Luxembourg, the Netherlands, Norway, Portugal, Spain and
Sweden. In the USA, the Environmental Protection Agency (EPA) has proposed
new emission limits for several categories of pulp, the so called cluster rules. The
proposed emission limit for AOX for bleached kraft pulp is 0.156 kg/t. The new
regulations will affect every existing and new facility in the pulp and paper
industry and are expected to be promulgated soon. An AOX limit of 0.156 kg/t
as proposed by the US EPA is very stringent when compared to current
emissions even from Scandinavian mills (Fig. 1). Germany is also discussing
legislation which will not only ban production of pulp using chlorine-containing
chemicals but also consumption of other pulps than totally chlorine free (TCF).
The Ministry of Environment and Forest, Government of India, has categorized
the pulp and paper industry as one of the twenty most polluting industries and
advised the paper industry to self-impose the AOX discharge limit of 2 kg/t of
paper. Some of the state pollution control boards in India have already intro-
duced the AOX limit of 2 kg/t of paper as a controlling parameter.
1.4
1.2 ] Scandinavian mills
II Canadian rnitis
1.0
0.8
v 0.6
x
o
Cluster rulesl
M
0.4
II
0.2
R
3456 10 11 1 2 1 3 1 4 1 5
I I I
161718192021 22,23:425 26 27 28 29 30 31
Mill
Fig. 1. AOX emissionsfrom Scandinavianand someCanadianbleachingkraft pulp millsin 1994

Despite some shortcomings, the kraft process is the most cost effective,
versatile and efficient wood delignification method available. Kraft pulp is
generally more difficult to bleach than sulfite pulp. With softwood pulp, it is
harder to achieve a high brightness product than with hardwood. Chlorine is
one of the popular bleaching chemicals, reacting with most of the lignin remain-
ing in pulp, and degrading and dissolving the lignin as chlorinated organics. The
acidic chlorination is usually followed by alkaline extraction to increase the
solubility of chlorinated lignins. The substitution of organically bound chlorine
by hydroxyl ions takes place under alkaline conditions, eliminating 60-90% of
organically bound chlorine in the pulp I-2]. The stages following the chlorina-
tion and alkaline extraction are known as bleaching stages, and use more
powerful oxidizing chemicals such as chlorine dioxide, hydrogen peroxide and
hypochlorite. The main reaction is to oxidize the chromophoric structures in the
pulp and to increase the pulp brightness. In the production of softwood kraft
bleached pulp by the conventional CE1DIE2D2 sequence, approximately 75%
of the dissolved material (COD and colour) and 95% of the organically bound
chlorine were contained in the C and E1 prebleaching effluents. The major
source was the E1 stage effluent. Lindberg 1-3] reported that the E1 effluent,
which constituted about 12% of the total bleach effluent, accounted for 96% of
the colour, 70% of the COD, and 50% of the BOD in whole bleach plant
effluent.
The amount of TOC1 produced during pulp bleaching varies with wood
species, kappa no. of pulp, bleaching sequence and conditions employed.
Typically, TOC1 in the effluent from a bleached softwood kraft pulp by a con-
ventional sequence is 5-8 kg/t of pulp bleached, representing about 10%o of the
total chlorine charged in the chlorination stage [4-6]. A physical-chemical
classification of this chlorinated organic material, present in spent liquors from
conventionally pulped and bleached softwood kraft pulp, is shown in Fig. 2
[7 9].
About 20% of the organically-bound chlorine found in the bleaching effluent
corresponds to relatively low-molecular-mass (M r < 1000) material. In recent
years, considerable research effort has been directed towards characterizing this
fraction with respect to its individual chlorinated compounds [10-12], as this
fraction is expected to contain those compounds which are potentially toxic to
aquatic organisms because of their ability to penetrate cell membranes or their
propensity to bioaccumulate in the fatty tissues of higher organisms. Some of the
major components of this low-molecular-mass fraction were found to consist of
relatively water-soluble substances such as chlorinated acetic acids or chlorin-
ated acetones, which are easily broken down [8, 9] before or during biotreat-
ment and are thus of minimal environmental significance. The fraction of AOX
which is extractable by a non-polar organic solvent and is referred to as EOX (or
extractable organically-bound halogen) accounts for about 1-3 %0 of the total
organically-bound chlorine. This fraction contains relatively lipophilic (i.e. fat-
soluble) neutral organic compounds, primarily of low molecular weight, and is
therefore of greater environmental significance than the remaining 99 %0 or so of
Reduction of OrganochlorineCompoundsin BleachPlant Effluents 219
8o% I
High M r material I
Relatively hydrophilic (Water soluble)
mainly non-aromatic
Does not permeate cell walls
I AOX < 10% chlorine by weight
100%
~ i R ~19% I
elatively hydrophilic
ncludes compounds
Low M20m~ which can easily be
hydrolyzed or metabolized
e.g. trichloroacetic acid)
Relatively lipophilic Log Pow > 3

(fat soluble) Highly lipophilic
Potentially toxic bioaccumulable
potentially bioaccumulable (e.g. Dioxin - 44%
chlorine by weight)
Fig. 2. The character of AOX in the effluentfrom conventionallypulped and bleached kraft
pulp
the AOX material. The EOX material can be further fractionated according to
its octanol/water partition coefficient (Pow). The fraction having a partition
coefficient greater than 1000 (i.e. log Pow > 3) makes up only 0.1% (or less) of the
AOX and contains those compounds which are most readily bioaccumulable
and considered to be potentially the most toxic and persistent. A component of
particular concern in this fraction is the polychlorinated aromatic material that
has a relatively high level of chlorine substitution, typically three or more
chlorine atoms per aromatic ring.
The major bleaching parameters such as incoming kappa number, C12
dosage, and chlorination and extraction pH and temperature have a significant
effect on the effluent BOD, COD, color loadings and the formation of chlorin-
ated compounds. It has been reported that the pollution load and amount of
chlorinated material produced in the chlorination and extraction stage are
a function of the amount of chlorine applied to the pulp, which is determined by
residual lignin in the pulp. The amount of TOC1 in the C stage spent liquor was
strongly dependent on the C12 dosage and lignin content in the pulp [13-15].
The lipophilic chlorinated organic substances were found to increase with an
increase in the pulp kappa number and C12 dosage [15]. An increase in the
chlorine dosage results in increasing BOD and COD in the chlorination and
extraction effluents [16, 17]. Voss et al. [4, 17] have reported that significant
reductions in the loadings of toxicity and chlorinated phenolics of combined
C and E effluents can be achieved by a higher final pH of chlorination. The
formation of TOC1 and chlorinated phenolics increased with increasing temper-

ature and decreasing pH of chlorination. Alfthan et al. [16] reported that
increase in temperature of the chlorination resulted in increase in pollution
loadings in chlorination and extraction effluents. Crawford et al. [18] reported
that the increase in the end pH and temperature of chlorination and chlorine
charge increases the formation of chloroform, potential mutagen and carcinogen
in the chlorination liquor. The hypochlorite stage is the major source of
chloroform formation. In the C stage, chloroform production is normally much
less, but it increases with increasing pH, temperature and kappa factor. The
higher kappa number also results in an increase in chloroform concentration in
the E stage. Chan and McDonald [19] reported that the use of high C102
substitution in the C stage reduced the chloroform concentration. While C102
substitution reduces the effluent TOC1, color and chloroform, it had little impact
on the production of chlorinated dioxins [20]. The major source of chlorinated
dioxins was found to be the C stage followed by the E stage. The formation of
dioxins is mainly dependent on the brown-stock kappa number, kappa factor,
mixing condition in the chlorination, carry-over and wash water quality.
This article presents a state-of-the-art review of the literature relating to the
environmental impact of the chlorinated organic compounds and the measures
which are used for their reduction. Emphasis is laid upon biotechnical methods
for reducing the discharge of chlorinated organics.
2 Environmental Impact of Chlorinated Organic Compounds
About 300 different chlorinated organic compounds in bleached pulp mill

effluents have been identified to date. About 200 of these are compounds which
include chlorinated resin acids, chlorinated phenolics and dioxins [20-22]. The
main compounds of the general type are listed in Table 2. These compounds,
classified as acidic, phenolic and neutral compounds, are at least partly respon-
sible for the oxygen demand (BOD and COD), effluent color, toxicity,
mutagenicity and carcinogenicity [23-29]. Various chlorinated phenolic, acidic
Table2. Chlorinated organic compounds in bleached pulp mill effluents

Type Number of varieties Amounts(g/t pulp)
Chlorinated phenolics 40 Up to 100
Chlorinated aldehydes,ketones and lactones 45 500
Chlorinated acids 40 Up to 500
Chlorinated hydrocarbons 45
Chlorinated ethers 20
High molecular mass Up to 4 kg C1
Based on data from Refs. [2~22]
Reduction of Organochlorine Compounds in Bleach Plant Effluents 221
and neutral compounds and chlorinated dioxins have been found to be bioac-
cumulative [20, 30]. Up to 2000 ppm organic chlorine has been detected in the
fat of fish from waters receiving bleaching effluent [31]. Untreated pulp and
paper mill effluents can be acutely toxic to fish at concentrations as low as 2%
by volume [26]. Chlorinated phenolics, resin and fatty acids are the principal
contributors to effluent acute toxicity [23, 32-34]. Table 3 lists 12 poly-
chlorinated phenolics selected by the US EPA for possible regulation [35]. The
toxicity of a chlorinated compound increases with increasing number of chlorine
atoms on the organic compounds. Polychlorinated dibenzodioxins (PCDDs)
and 2,3,7,8-tetrachlorinated dibenzodioxin (TCDD) are toxic. It is generally
believed that they are of precisely the right size to fit as the key in the lock in
some vital molecules in living cells and to be firmly attached to this site by the
chlorine atoms at both ends, thus preventing normal functioning of the cells.
T C D D seems to fit perfectly. If, however, more hydrogen atoms in the benzene
rings are substituted with chlorine, the toxicity is reduced, e.g. if all eight
available hydrogen atoms are replaced with chlorine atoms, the toxicity drops to
one per thousandth of that of TCDD. The key does not fit in the lock any more.
Dioxins have received extensive media attention. 2,3,7,8-TCDD is extremely
toxic and bioaccumulative. The toxicity of individual members of the above-
mentioned family of 210 isomers (varieties) of polychlorinated dioxins and
furans differ substantially. The most toxic of them is at least 100000 times as
toxic as the least toxic, so that it is meaningless to judge the quality of an effluent
or pulp by quoting the total dioxin concentration. Dioxins are frequently
reported as toxicity equivalents (TEQ), which is the sum of total chlorinated
dioxins and furans corrected for the toxicity of each relative to 2,3,7,8-TCDD.
For practical purposes, in the pulp industry, the T E Q generally corresponds to
somewhat more than the concentration of 2,3,7,8-TCDD [36, 37].
Table 3. Polychlorinatedphenoliccompoundsproposed for regulation

by the US EPA
Polychlorinatedphenols Minimum level,ppb (gg/1)"
Pentachlorophenol 5.0
2,3,4,6-Tetrachlorophenol 2.5
2,4,5-Trichlorophenol 2.5
2,4,6-Trichlorophenol 2.5
3,4,5-Trichloroguaiacol 2.5
3,4,5-Trichlorosyringol 2.5
3,4,5,6-Tetrachlorocatechol 5.0
3,4,6-Trichlorocatechol 5.0
3,4,5-Trichlorocatechol 5.0
3,4,5,6-Tetrachloroguaiacol 5.0
"The minimum level is defined as the concentration at which the
analytical system gives recognizablemass spectra (correctedfor back-
ground) and acceptable calibration points, using EPA method 1653
Greenpeace claims that there is no safe level of dioxin [36]. On the other
hand, over a billion dollars worth of research has failed to prove any serious
health damage to humans due to dioxin exposure [36].
It is fairly well accepted by scientists in the field that 10 picograms of dioxin
per kg body weight per day is an acceptable daily intake (ADI) for a one in
a million risk of mortality, over and above the other risks of living. However,
a concern about dioxins in effluents is that they bioaccumulate in fish. This
means that they concentrate as they move up the food chain through predator
fish to humans. In Canada, the Federal Government recommends that fish with
over 20 ppt dioxins in the flesh should not be eaten. Occasional fish caught
downstream of mills have concentrations of up to about 100 ppt, but these
incidents can be expected to become things of the past if mills adopt the dioxin
control measures already announced. To help put this all into perspective, it is
worth noting that collection of the ADI includes several safety factors. Extensive
studies of 1200 US soldiers who worked with dioxin-contaminated herbicides in
Vietnam failed to demonstrate any health effect, but there was an out-of-court
settlement of almost $200 million. One might say that legal dangers have been
demonstrated to be associated with dioxin but no serious biochemical danger to
humans.
Recently, the EPA released its 2000-page draft reassessment of the environ-
mental and health effects of dioxins [38]. The report reaffirms the EPA's 1985
conclusion that dioxins and related chemicals are a probable cause of cancer in
humans. It also presents new evidence that dioxins, even in trace amounts, may
cause a wide range of other adverse human health effects including disruption of
regulatory hormones, reproductive and immune system disorders and abnormal
fetal development. The EPA defines dioxins and related compounds as tetra- to
octa-chlorodibenzo-dioxins and furans with chlorine atoms in at least the 2,3,7
and 8 positions, as well as coplanar polychlorinated biphenyls (PCBs). The EPA
expresses the mass of these compounds in terms of their toxicity compared with
the most toxic dioxin congener 2,3,7,8-tetrachlorodibenzo p-dioxin, denoted as
2,3,7,8-TCDD toxicity equivalents (TEQs). In other words, if the mass of
a particular dioxin congener is 100g and it has one tenth the toxicity of
2,3,7,8-TCDD, its mass in TEQs is 10 g.
The reassessment emphasizes dioxin's effects on fetal development, because
these effects have been seen in the offspring of laboratory animals exposed to
very low concentration, and are now being observed in children accidentally
exposed to dioxin-like compounds in the womb [38]. For example, the offspring
of women who in 1979 ate rice oil contaminated with furans and PCBs in the
Yu-Cheng Province of Taiwan exhibit changes in skin pigmentation and hair
growth and are showing signs of abnormal sexual development, says Aronold J.
Schecter, Professor of Preventive Medicine at the State University of New York,
Clinical Campus, Binghamton. Dioxins and dioxin-like compounds apparently
target many sets of genes which encode a variety of proteins, including hor-
mones, enzymes and growth factors. As a consequence, the cell produces an
inappropriate amount of protein. Depending upon the dose, timing and age of
the individual, this cell disruption can lead to diverse biological outcomes,
effects on the reproductive system of the developing fetus, effects on the brain,
disruption of the immune system and cancer. Dioxin's effect has been seen in all
species studied, although they occur at different doses. Humans lie somewhere in
the middle of the sensitivity range, neither extremely responsive nor extremely
resistant, according to the EPA report. Unlike natural hormones, like the
estrogen estradiol, dioxins can be deactivated by binding to sex-hormone-
binding globulin and are not easily metabolized. Dioxins disrupt multiple
endocrine systems, at times behaving as antiandrogens, at others like antiestro-
gens or even estrogens. The half-life of the most studied dioxin 2,3,7,8-TCDD is
10 years in humans [39].
Low molecular mass chlorinated neutral compounds are major contributors
to mutagenicity [31]. The dominant chlorophenolics in C stage effluent are
chlorocatechols, whereas chloroguaiacols are the major species in E stage
effluent. In hardwood bleaching effluent, additional chlorinated compounds
such as chlorinated vanillins, syringealdehydes and syringals were found. But
the concentrations of chlorinated phenolics in the hardwood bleaching effluents
are generally lower than those in softwood bleaching effluent [40]. At least 14
different chlorinated phenolics in conventional C and E1 bleaching effluents
were detected [41].
Sublethal effects of pulp and paper mill effluents are varied. The threshold
concentration for sublethal effects appears to be near 1/10 of the 96 h LCso
concentration [33]. Davis [42] reported the lethal effect at 5 % of the 96 h LCso
concentration of the spent bleaching liquor.
The Swedish pulp and paper industry sponsored studies to determine the
environmental effect of spent bleach liquors and to develop environmentally
compatible bleaching processes [15, 43, 44]. It was concluded that the effects of
bleach plant effluents actually observed in receiving waters were few in number
and limited to receiving waters with poor water exchange or to areas close to the
outlet. Laboratory tests suggested that all sublethal effects on fish of effluents
from the conventional bleaching of softwood kraft pulp disappear after a total
dilution of 70-1000 times [15]. Based on the results of chemical and biological
comparisons of the effluents, various alternative bleaching sequences and treat-
ments were ranked as shown in Table 4. The Swedish project SSVL-85 was
designed to assess long-term large-scale effects of bleach plant effluents and used
model ecosystems to characterize bleach plant effluents [15, 44]. Two concen-
tration levels were used corresponding to dilutions of 400 and 2000 times for
a total effluent volume of 50 m3/t pulp. Five different effluents were tested for
fish and invertebrate toxicity, reproductive disturbances, physiological changes
and disease, parasitic infestation, bioaccumulation of chemicals and genotoxic
effects (mutagenicity and/or carcinogenicity). The ranked sequences did not
change significantly from that in Table 4. However, biological effects were found
at dilutions which were previously thought to render the effluents harmless. The
effluent from a conventional (C95D0s)E1DE2D bleach plant showed strong
biological effects after 166-fold dilutions, with low survival of fish, decreased
Table 4. Bleaching processes in order of decreasing environmental effect
S.no. a Bleaching process Comments
1 (C90DIo) E1HD1E2D2 Traditional technique

2 (CgoDIo) E1HD1E2D2 Improved washing compared to
traditional technique
3 (C9oDlo)E1HD1E2D2 + UfE1 Ultrafiltration of Ex effluent
4 0(C85D15) E1D1E2Dz Oxygen bleaching to kappa no. 20
and washing to 1~12 kg COD/t
5 (C9oDao)EaHD1E2D2 + UfE1 + Z Ozone treatment of permeate and
C-stage effluent
6 0(C85Dxs)E1D1E2D2 Oxygen bleaching to kappa no. 20
and washing to 4-6 kg COD/t
7 (C90Dlo)E1HDaE2D2 + aerated lagoon Total effluent through aerated lagoon
8 (CgoDlo) + Ion exchange Ion exchange of total effluent
9 (D85C15)EID1E2D 2 High substitution of chlorine dioxide
10 0(C85D15)E1D1E2D2 + aerated lagoon Oxygen bleaching to kappa no. 20
11 0(CssD15)E1D1EzD2 Oxygen bleaching to kappa no. 17
aSoftwood Kraft Pulp

UfE~ = Ultrafiltration of E1 effluent
Worst 1
Best 11
Based on data from Ref. 1-15]
invertebrate density and parasitic infestation of stationary fish species. Even at

dilutions over 5000 times, all these effects could be found, although to a substan-
tially decreased degree.
In 1983, the Swedish Government sponsored Project Environment/Cellu-
lose, a three year study of the Gulf of Bothnia, that portion of the Baltic sea
which lies between Sweden and Finland [21]. The study focussed on the area
around the Norrsundet pulp mill, which discharges untreated effluent into the
Gulf and which was regarded as representative of Swedish pulp mills. Near the
discharge point, fish biomass was low, species composition was changed, repro-
duction was reduced and physiological disturbances were seen. The effluent also
affected the diversity, biomass and distribution of invertebrates and plants in the
region [21]. The toxic effects were more pronounced within 4-5 km of the outlet
of the mill, but biological effects were observed in the fish caught 8 to 10 km
from the outlet, where the dilution was estimated to be 5000 times [45].
Bioaccumulating compounds in the fish declined in concentration from the
discharge point to about 5 km out from the shore [21]. The concentration of
AOX in the water, however, showed a larger area of influence. High concentra-
tions of solvent-extractable organically bound chlorine (EOC1) were found in
the sediment 15 km out from the Norrsundet mill and 29 km out from another
mill 75 km north of Norrsundet. Large areas of the Gulf of Bothnia were found
to have higher levels of EOC1 in the sediment. Compared with areas further from
industry, EOC1 levels in sediment 20 to 50 km off the coast of bleached kraft
pulp mills were 10 times the background levels [21]. Between 1979 and 1984, the
Norwegian Centre for Industrial Research studied the content of EOC1 in fish in
relation to the effluent of a CEHD bleached sulphite pulp mill [46]. While the mill
was in operation, the concentration of EOC1 in fish near the outlet was up to 30
times that in fish from a nearby reference area. EOC1 levels in fish did not drop to
the background concentration until 3.5 years after the mill had shut down.
In 1987, Swedish scientists stated that in spite of extensive investigations,
a clear-cut relationship between release of chlorinated organic material and bio-
ecological effects in the receiving waters has not yet been established. Thus, the
TOCI regulations give a general feeling that the release of chlorinated organic
material from bleach plant is critical to the environment [47]. However, concern
has been voiced that the effects found in Sweden are specific to that area,
because the Baltic sea may be a unique ecosystem. It contains a relatively small
number of species and the media is brackish and has a long residence time, of the
order of several decades [483.
Extensive long-term environmental impact studies have also been carried
out in the United States. Since virtually all the mills in the USA have secondary
biological treatment, treated bleach plant effluents were tested. Test work, using
experimental stream channels, has been conducted by National Council of the
Paper Industry for Air and Stream Improvement (NCASI) as part of an aquatic
biology investigation going on since the 1970s. In the latest report, the results of
year-long exposure to treated bleached plant effluent of 20:1 dilution were
reported [49]. Productivity of the rainbow trout population and aquatic flora
was increased compared to that in the control stream, while productivity of the
benthic macroinvertebrate population was unchanged. There was no effect on
the diversity index or histopathology of 20 different fish-tissue types. These
results are clearly contradictory to the general conclusion of the Swedish
investigations. The reason for the differences is not apparent; it may be the effect
of effluent treatment or the sensitivity of the Baltic eco-system.
Another category of compounds which has aroused environmental concern
comprises the high-molecular-mass chlorinated compounds (Mr > 1000). In
bleaching softwood kraft pulp, this fraction of chlorinated compounds accounts
for 50% of dissolved organic material [50] and 70% of TOC1 [20]. Although
these compounds contribute little to BOD5 and acute toxicity due to their
inability to pass the bacterial cell membrane, they are the major contributors to
effluent colour, COD and chronic toxicity. Eriksson and Kolar [51] and
Eriksson et al. [52] found that the high-molecular-mass material was not as
stable as previously thought. High-molecular-mass chlorolignins from pre-
bleaching stages were chemically unstable under conditions that may prevail in
receiving waters. The material was slowly decomposed to various chlorinated
catechols and guaiacols which were methylated in cases where a complete
mixture of bacteria or the white rot fungus alone was used, a condition appear-
ing to be common in nature [15, 53]. The low-molecular-mass phenolics and
their methylated counterparts (more lipophilic) may cause toxicity and bioac-
cumulation in fish.
Chlorate is yet another pollutant recently arousing environmental concern.
The formation of chlorate is largely dependent on the amount of C102 used in
the bleaching. It is a well-known herbicide and biocide, especially for brown

algae [54]. A conventional bleaching sequence produces about 3-6 kg/t of pulp
bleached depending on the level of C102 substitution.
3 Reducing the Generation of Organochlorine Compounds
The emission of chlorinated organic compounds from bleach plants can

be reduced by modifying the pulping and bleaching processes using one or
more of the following strategies: (1) removing more lignin before starting
the chlorination, i.e., reducing the kappa number of unbleached pulp, (2)
modifying the conventional bleaching process, (3) eliminating all chlorine
compounds in bleaching, or (4) recovering bleach plant effluents. These
methods may be physicochemical or biochemical in nature or a combination
thereof.
Lignin from the pulp before chlorination can be removed by extended
delignification of the brown stock pulp using modified continuous cooking
(MCC) [55-58] or modified batch cooking (MBC) [59-61]. The lignin content
can also be removed by oxygen delignification [20, 58, 6~69]. Ozone is also
capable of delignifying an oxygen-treated pulp to a very low kappa number of
5-6 [70, 71]. A lower lignin content in the pulp requires lower chlorine usage
resulting in reduced generation of chlorinated organics. The conventional
bleaching process can be modified by (a) reducing the kappa factor or active
chlorine multiple, i.e., the total active chlorine charge derived by the kappa
number of the pulp entering the chlorination stage, (b) slow and multiple
addition of chlorine [72], (c) chlorine dioxide substitution in the chlorination
stage [4, 13, 17, 72-77], (d) oxidative extraction using oxygen [78-80] and/or
hydrogen peroxide [81-91], or (e) converting to ECF bleaching sequence
[89, 92]. Further decrease in AOX can be achieved by a combination of
extended delignification in the digestor, oxygen delignification, and high chlor-
ine dioxide substitution in the chlorination stage [72]. The trend now is to adopt
a totally chlorine-free (TCF) bleaching sequence in some European countries or
ECF bleaching in North America, which can involve the use of oxygen, enzyme,
hydrosulfite, peroxide, peracetic acid, thiourea dioxide, chelating agents
[72, 93], ozone [70-72, 94-97] etc.
Once chlorine-free bleaching is introduced, the bleaching effluent can be
taken to the black liquor recovery section [98]. A non-polluting bleach plant
has been reported where counter-current washing decolorization and reuse of
the extraction stage effluent as wash water in the chlorination stage is practiced
[99, 100]. Ultrafiltration and reverse osmosis membrane techniques can also be
applied for complete recycling of the effluent [101, 102]. The best-known closed
cycle effluent-free bleaching process was reported in the mid-1960s by Rapson
and Reeve [103-105].
Reduction of Organochlorine Compoundsin Bleach Plant Effluents 227
Apart from the above methods, several biochemical and biological methods
can be used to reduce the generation of organochlorine compounds, as discussed
in the following sections.
3.1 Enzymatic Prebleaching
Pretreatment of pulp with enzymes prior to bleaching helps in substantially

reducing or eliminating the use of chlorine and chlorine compounds. Hemicel-
lulase enzymes and ligninolytic enzymes have been investigated. Hemicellulase
enzymes have no immediate effect on the kappa number and lignin removal of
the treated pulp. However, the consumption of the bleaching chemicals is
reduced and/or brightness ceiling is increased. Ligninolytic enzymes selectively
remove the lignin from the pulp fibre, hence are more useful. The role of different
enzymes on pulp components has been reviewed by Eriksson [106].
3. I. 1 Hemicellulase Enzymes
These enzymes are used commercially in pulp bleaching. The main enzyme
needed to enhance delignification of kraft pulp is reported to be endo-~-
xylanase, but enrichment of xylanase with other hemicellulolytic enzymes has
been shown to improve the effect of enzymatic treatment [107-110]. Xylanases
act mainly on the relocated, reprecipitated xylan on the surface of the pulp
fibres. Enzymatic hydrolysis of this specific type of xylan renders the structure of
the fibre more permeable, allowing lignin and lignin carbohydrates to diffuse
more easily into the bleaching liquor. An alternative explanation is that
xylanases attack the bonds that exist between xylan and lignins, releasing lignins
which can then diffuse more easily into the bleaching liquor [109, 111, 112].
Xylanase treatment has been shown to reduce the requirement of chlorine
for bleaching, while still achieving high brightness and good pulp properties
[113-115]. Results from laboratory study and mill trials show about 3 5 4 1 %
reduction in active chlorine at the chlorination stage for hardwoods and
10-20% for softwoods, whereas the saving in total active chlorine is found to be
20-25% for hardwoods and 10-15% for softwoods [116-123].
Xylanase treatment represents a successful new technology for reducing
chlorine use. More details on xylanase bleaching are available in a recent review
by Bajpai and Bajpai [124] and in another chapter in this volume written by
Viikari et al. [125].
3.1.2 Ligninolytic enzymes
These enzymes, unlike xylanases, attack lignin directly, and hence are more
effective. White-rot fungi are the main producers of ligninolytic enzymes. These
fungi secrete a number of oxidative enzymes and some hitherto unknown

substances (mediators) into their environment, together effecting a slow but
continuous degradation. The most important lignin-degrading enzymes are
lignin peroxidases, manganese peroxidases and laccases. The action of both
lignin peroxidases and manganese peroxidases needs Mn z +, which is oxidized to
Mn 3 +. Mn a + is the real oxidizing agent, attacking the lignin molecule. Laccase
uses molecular oxygen as a cosubstrate. In the absence of the living organism,
the various peroxidases and laccases perform only negligible kappa number
reduction [126, 127]. Egan [128] has reported kappa number reductions of 24
and 26% after treatment with ligninase 118 from P. chrysosporium followed by
alkaline reduction. Viikari et aI. [129] were unable to demonstrate any bleach-
ing effect on pine kraft pulp with ligninases and oxidase from the white-rot
fungus Phlebia radiata or purified ligninase from P. chrysosporium. When the
ligninases were applied after treatment with hemicellulases, there were slight
reductions in kappa number. Arbeloa et al. used lignin peroxidase enzyme from
P. chrysosporium to improve the bleachability of hardwood and softwood kraft
pulps [130]. Enzyme treatment prior to chemical bleaching increased brightness
and decreased lignin content in the pulp. The final brightness of pulp was found
to be higher by about 0.84).9 points than that of the control (Table 5).
Although much of the research has focussed on lignin peroxidases, these
enzymes are not necessarily involved in lignin degradation and may not be
secreted by all lignin-degrading fungi. C. versicolor produces laccases as well as
lignin and manganese-dependent peroxidases [131-134]. However, Archibald
[135] recently found that lignin peroxidases secreted by C. versicolor did not
appear to play an important role in lignin degradation. Dichomitus squalens
[136] and Rigidoporus lignosus [137] produce both laccase and a manganese-
dependent peroxidase but do not produce lignin peroxidase.
It is now known that although lignin peroxidases and laccases play an
important role in degrading the lignin in vivo, in vitro the oxidation reactions
catalyzed by the enzyme result in further polymerization of the lignin [138].
Table 5. Effects of lignin peroxidase on kappa number and bright-

ness in softwood and hardwood kraft pulp
After treatment After bleaching
Kappa Brightness Brightness

number (%) (%)
Softwood
Control 29.7 29.1 88.8
Treated 26.9 24.1 90.7
Hardwood
Control 14.6 31.0 88.6
Treated 11.8 39.0 89.4
Conditions of enzyme treatment: pH 3, temperature 30 ~ veratryl

alcohal 2 mM/l, H202 addition at 100 ppm/h for 3 h
Recently, Hammel and Moen [139] reported depolymerization of a synthetic

lignin by a lignin peroxidase in the presence of H202 and veratryl alcohol, but
this effect has not been demonstrated with lignin in wood or pulp. It is likely that
fungi possess enzyme systems that prevent polymerization [138, 140].
These results imply that single enzymes are not able to mimic the complete
biological system. Small improvements can be achieved by the addition of
low-molecular-mass aromatic compounds like veratryl alcohol or other sub-
stances such as ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonate)] and
Remazol blue [126, 127]. In consequence, most scientific and industrial groups
have meanwhile withdrawn from research programs focussing on direct lignin
degradation.
Lignozyme GmbH (Germany) continued work with enzymes plus chemical
mediators which create a redox system throughout the pulp treatment period
[-141-143]. Their idea was to find a system which is a good mimic of the natural
situation. Starting in 1987 with the enzyme mediator concept, Lignozyme has
very recently improved the performance of the mediator system for the laccase of
Coriolus versicolor by changing and further fine-tuning the chemical nature of
the component [144-146]. The treatment of pulp with laccase alone does not
result in any degradation of lignin but merely produces in a structural change or
repolymerization, whereas the laccase mediator system causes a significant
kappa number reduction at reasonable treatment times even if the enzyme
mediator system is applied in several consecutive treatment steps to the same
pulp. In contrast to commonly used pulp-bleaching chemicals (oxidizing and
reducing), no passivation can be observed in enzymatic delignification, i.e.,
efficiency does not drop significantly during consecutive treatment steps. Lac-
case enzyme contains four atoms of copper per molecule and requires oxygen as
a cosubstrate for the oxidation reaction which has to be provided. According to
the present understanding, the laccase, while oxidizing the chemical mediator, is
generating a strongly oxidizing co-mediator which is the real bleaching agent.
Table 6 illustrates the conceptual difference between the indirect and the direct
enzymatic lignin attack. The general technical conditions for enzymatic bleach-
ing with the laccase mediator system are: temperature 40-65~ pH 4-7,
consistency 1-20%, pressure 1-14 bar, duration 1~4 h. The system provides
a broad flexibility with respect to the pulp substrate, the technical requirements
for application, and the final quality of the pulp. The principal applicability has
been demonstrated for softwood and hardwood pulps as well as for annual plant
Table 6. Differencesbetweenxylanaseand laccase/mediatortreatment

Xylanase Laccase/mediator
No or verypoor kappa reduction Very good kappa reduction
Moderate bleachingeffect Good bleachingeffect
Saving of bleachingchemicals Saving of bleachingchemicals
TCF pulp productionpossible
fibres. A repeated enzymatic treatment is possible and results in a 50-70%

kappa number reduction per treatment step. Laccase mediator system (LMS) is
compatible with all other bleaching sequences. The performance of the LMS
(1-hydroxybenzotriazole) has been proven in several pilot trials and is now
ready for large-scale application.
Recently, Kondo et al. [147] examined the bleaching of kraft pulp with Mn
peroxidase (MnP) enzyme, which was isolated from cultures of P. sordid YK-
624. Pulp brightness was increased by about 15 points and kappa number
decreased by about 8 points when unbleached kraft pulp was treated with MnP
in the presence of MnSO4, Tween 80 and sodium malonate with addition of
H 2 O 2 at a rate of 3 ml/h at 45 ~ for 12 h. To establish an absolutely chlorine-
free bleaching process, oxygen-bleached kraft pulp (OKP) was treated with
a four-stage biobleaching process consisting of sequential MnP treatment,
alkaline extraction, MnP treatment and hydrogen peroxide treatment. Fully
bleached kraft pulp (brightness 91%, yield 97%) could be obtained from OKP
by combination of enzyme treatment and hydrogen peroxide bleaching.
MnP from Trametes versicolor was found to delignify softwood kraft pulps
over a wide range of initial lignin contents [148]. Demethylation of residual
lignin in the pulp also occurred. Subsequent treatment of pulp with TCF
chemicals resulted in brighter pulp than predicted from the initial kappa
number. The MnP stage required small doses of hydrogen supplied by the
action of glucose oxidase on glucose. Mn ions already present in the pulp were
chelated by malonate or gluconate during the catalytic cycle. Much of the
bleaching effect was evident after 4 h of treatment with 1 U/ml of MnP, although
bleaching was improved further after 24 h under the conditions used. Laccase,
especially in the presence of the mediator, ABTS, produced a similar effect to
MnP when combined with subsequent TCF bleaching. MnP generates o-
quinones by demethylation of residual lignin, and these structures are suscep-
tible to oxidative ring opening during TCF bleaching, resulting in hydrophilic
soluble lignin.
Ducka and Pekarovicova [149] used crude ligninases from P. chrysosporium
for bleaching of softwood kraft pulp. The pulp, after oxygen bleaching with
kappa number 19.7 and 36.5% MgO brightness, was pretreated with ligninases
(L) or xylanases (X) and bleached in QEopDP sequence. The brightness of pulp
bleached in this sequence was 87.8%, which is about 3.2 points higher than the
control and is approximately the same as that obtained with the use of commer-
cial xylanase.
3.2 Fungal Prebleaching
Pretreatment with fungi has been shown to replace up to 70% of the chemicals
needed to bleach kraft pulp [150]. The usual specificity of biological reactions
and their mild reaction conditions make biological delignification an interesting
alternative to bleaching with chemicals such as pressurized oxygen or ozone.
Only a few white-rot fungi have been tested for their ability to delignify kraft
pulps. Phanerochaete chrysosporium reduced kappa number by about 33% in
hardwood kraft pulp during a 10 day incubation period, whereas Coriolus
versicolor reduced kappa number by about 20 and 33% in a 5 day period
[151-153]. Table 7 shows the performance of various white rot fungi used in
treating hardwood kraft pulps. The largest bleaching effect was noted
with white-rot species IZU-154 [154]. Treatment of hardwood pulps
with alkaline extraction following fungal bleaching did not significantly improve
brightness. Softwood kraft pulps were found to be more resistant to attack by
P. chrysosporium and C. versicolor, possibly because both P. chrysosporium
and C. versicolor tend to attack hardwoods more often than softwoods in
nature [155-157]. Softwood lignin has a different character from hardwood
lignin and is susceptible to blocking reactions that restrict delignification
[158, 159].
On delignification of softwood pulps by P. chrysosporium, kappa number
reductions of 50-70% were achieved [160, 161]. Kirk and Yang [160] reported
a considerable loss in cellulose content during fungal bleaching, which
was attributed to cellulase production in the strain being used. A strain of
P. chrysosporium without cellulase activity has since been developed [162].
Table 7, Change in brightness of hardwood kraft pulp after exposure to various

white-rot fungi
Brightness (%ISO) Incubation
Period
Fungus Increase Final (d)
Phanerochaete 11 39 13
Chrysosporium 0 37.7 14
2.2 35.7 5
P. chrysosporium
(cellulase-free mutant # 431) 2.0 35.5 5
P. chrysosporium
(cellulase-free mutant # 432) 1.9 35.4 5
Coriolus versicolor 8 25 50-67 10
7.1-17.8 38.5 50.3 5
14.5 48.0 5
13.6 50.4 5
13.4a 50.2 5
6 34.0 15
IZU-154 35 63 11
Coriolus hirsutus 23.7 52 5
12 40 11
Phellinus pini 2.3 35.8 5
Pleurotus eryngii 1.9 34.4 5
Pleurotus sajor-caju 2.3 35.8 5
Lentinus edodes 2.4 35.9 5
Aureobasidium puUulans 2.5 36.0 5
"Fungus was immobilized
Based on data from Refs. [152 154,163,195,199]
C. versicolor was found to be ineffective in brightening softwood kraft

pulps despite reductions in kappa number of 8% [163] and 47% [154, 157] over
14 days. However Reid et al. [157] reported that when the pulp was sub-
sequently subjected to alkaline extraction, brightness was increased by 15 points
and kappa number was further reduced. The softwood kraft pulp exhibited
a more pronounced darkening than the hardwood kraft pulp during the first
6 days of treatment, and a longer time lag occurred before kappa number began
to decrease. Direct brightening of softwood kraft pulp was noted when the
C. versicolor was immobilized on polyurethane foam, possibly because of a high-
er fungus/pulp ratio [157].
In Japan, a 5-day fungal (F) treatment of hardwood kraft pulp with IZU-154
replaced a CE1DEzD sequence with an FCED sequence, yielding respective
brightness of 88.8 and 88.1% ISO (84.2 and 85.3% ISO after aging) [154]. The
resulting chlorine saving was 72%, despite a kappa number reduction of less
than 60% in the fungal stage. In another study, a 5-day fungal stage with
C. versicolor essentially replaced the chlorination stage in a conventional bleach-
ing sequence, achieving a brightness of 82% ISO with a FDED sequence
compared to a brightness of 88% ISO with a CE1DEzD sequence [154].
Recently, Kondo et al. have reported biobleaching of hardwood kraft pulp
with lignin-degrading fungi P. sordia YK-624 [147]. A three-stage bleaching
process (5 days fungal treatment, alkaline extraction and again 5 days fungal
treatment) bleached the kraft pulp to about 80% brightness in 10 days, although
brightness increase of the pulp with a one-stage continuous fungal treatment for
10 days leveled off after 5 days incubation.
Wroblewska and Zielinsk [164] examined biodelignification of beech and
birch pulp wood by selected white-rot fungi. One of the strains, designated as
DL-Sth-4, was found to be the best for selective delignifiction of beech wood.
About 25% lignin was lost with very little loss in cellulose content. Pazukhina
et al. used the culture filtrate of several white-rot fungi P. sanguinea, C. ver-
sicolor, Ganoderma applanatum and Trichoptum biforma for bleaching hardwood
kraft pulp [165]. P. sanguinea showed the highest selectivity in lignin degradation.
Nishida et al. [166] investigated the biobleaching of hardwood unbleached
kraft pulp by P. chrysosporium and T. versicolor in the solid state and liquid
state fermentation systems with four different culture media (low nitrogen-high
carbon, low nitrogenqow carbon, high nitrogen-high carbon and high nitro-
gen-low carbon). In the solid state fermentation system with L N - H C culture
medium, pulp brightness increased by 15 and 30 points after 5 days of treatment
with T. versicolor and P. chrysosporium respectively. The pulp kappa number
decreased with the increasing brightness, and a positive correlation between the
kappa number decrease and brightness increase of the fungus-treated pulp was
observed.
Delignification by white-rot fungi is strongly influenced by culture condi-
tions. The major culture parameters affecting lignin degradation are: growth
substrate, nitrogen availability, other culture conditions, oxygen concentration
and mode of cultivation. P. chrysosporium and C. versicolor require a carbon
source such as glucose or cellulose to metabolize lignin to form carbon dioxide

[167], although the role played by the carbon source is not clear. Glucose
supplementation at low concentration (0.47%) was found to stimulate delignifi-
cation in kraft pulps [15!] and lignin in aspen wood [168]. However, there was
no difference in the degree of delignification for thermomechanical pulp (TMP)
at glucose concentrations ranging from 0 to 35% [169]. Glucose was added in
most studies to maintain pulp yield because it is a repressor of endo-l,4,fl-
gluconase, mannanase, xylanase, aryl-/%glucosidase, pectinase and cellulase,
which attack the carbohydrate fraction of the pulp. The white-rot fungus
IZU-154 was able to degrade lignin in hardwood kraft pulp without an
exogenous energy source and without significantly affecting pulp yield. Fusa-
rium solani and Aspergillus japonicus were also able to degrade lignin without
additional carbon source [154, 170-174].
Nutrient depletion, particularly nitrogen depletion, appears to trigger the
development of the ligninolytic system in P. chrysosproium, C. versicolor and
Phlebia brevispora, Phanerochaete sordida and Phlebia radiata with a few excep-
tions [152, 168, 175-179]. The effect of nitrogen depletion does not necessarily
hold true for all white-rot fungi, as the lignin-degrading abilities of P. sajor-caju
and L. edodes do not show strong stimulation by nitrogen depletion [180]. The
ligninolytic system of P. chrysosporium is also triggered by limitations in carbon,
sulphur, Zn z+, Fe 2+ and Mo 4+, but not phosphorus [30, 181, 182]. Other
culture conditions such as temperature and pH have not been studied extens-
ively [181]. Tran and Chambers [151] reported that the optimal temperature
for delignification by P. chrysosporium was 38 ~ but Drew and Kadan [183]
found that the extent of degradation of 14C-kraft lignin in 15 days at 28 ~ was
about 4 times that at 38 ~ Optimum pH values reported for lignin degradation
are different for different fungal strains. Tran and Chambers reported that the
optimal pH of P. chrysosporium was 3.5 for delignification and 4.5 for growth
[151]. Kirk et al. [184] reported an optimum pH of 4 4 . 5 for their strain.
Suppression of delignification was noted at pH less than 3.5 and greater than 5.5
[ 184]. Recently, Royle et al. reported increased lignin degradation by P. chrysos-
porium and L. edodes at pH 3 [180]. o-Phthalate was found to inhibit delignifica-
tion [185]; in one study, lignin degradation rates were doubled by changing the
buffer from o-phthalate to 2,2-dimethylsuccinate (DMS) [176].
Lignin degradation in white-rot fungi is primarily oxidative. It occurred
faster in cultures of P. chrysosporium which had been flushed with pure oxygen
than in those flushed with air [-135, 151,169, 186-188]. Enhanced lignin degra-
dation due to elevated oxygen partial pressures has also been reported for other
white-rot fungi including C. versicolor, Pycnoporus cinnabarinus, Lentinus
edodes, Giifola frondosa, Polyporus burmalus and Merulius tremellosus [189].
However, other species such as Gleoporus dichrous, Pleurotus ostreatus, Bondar-
zewia berkeleyi and IZU-154 were less responsive to increased oxygen concen-
trations [154, 189]. The addition of polydimethylsiloxane (PDMS) oxygen
carriers enhanced lignin degradation by C. versicolor [190]. However, they
alone were not found to appreciably increase the brightness of hardwood kraft
pulp [191]. Together with C. versicolor they acted synergistically, resulting in an

overall increase of approximately 10 brightness points [190].
Lignin degradation is also strongly influenced by the mode of cultivation.
Most researchers have studied stationary cultures, although dependence on
stationary cultures causes difficulties in scale-up. Also, agitation is an important
consideration because of the role of oxygen mass transfer. In a number of
studies, agitation in reciprocating and gyratory shakers caused the formation of
pellets consisting of mycelia entangled with pulp. The formation of mycelial
pellets resulted in suppression of degradation of kraft lignin and lignin model
compounds by P. chrysosporium [-151,156, 169, 192, 193]. Similar delignifica-
tion rates were noted in agitated and stationary cultures and also with the
addition of detergent to the culture [187, 194]. Effective delignification has also
been reported in agitated cultures of C. versicolor in which the formation of
mycelial pellets was prevented and in cultures of P. chrysosporium in the
presence of veratryl alcohol [195, 196, 199].
P. chrysosporium and C. versicolor have been successfully immobilized on
polyurethane foam [153, 157, 190, 197]. Immobilized and free cultures of C.
versicolor have been found to bleach hardwood and softwood kraft pulp at
a comparable rate and to a similar extent [153, 157]. The results showed that
intimate contact between the fungal hyphae and pulp fibres was not required
as long as the media was renewed through contact with the fungus
[132, 153]. Immobilization enabled the pulp to be separated from the mycelia.
Another advantage of immobilization was that the same fungal biomass could
be reused to treat other batches of pulp either immediately or after storage at
4~ [153].
A serious shortcoming of the fungal bleaching process is the long incubation
time required, ranging from 5 to 14 days for both hardwood and softwood pulps.
Softwood incubation periods are likely to be longer than those for hardwood
pulps because softwood pulps require a longer time lag (6 days) before kappa
number decrease or brightness increase can occur [157]. Hardwood pulps
generally have a lag time of 1-2 days [152-154, 160], followed by rapid and then
slower delignification.
Unfortunately, the size of the fungal bioreactor would have to be very large,
considering that daily production could range from 200 to over 1000 air-dried
tonnes of pulps. Most researchers have performed fungal bleaching experiments
at low pulp consistencies of 0.5-2% (w/v) [151 154, 190]. Only Fujita et al.
[154] have investigated fungal bleaching at high consistency (16-24%), which
would allow for a smaller reactor. Both reactor size and incubation period will
have to be reduced in fungal bleaching if it is to become an economical
alternative.
Jurasek and Paice [198] suggested that the lignin may become more flexible
and hydrophilic as a result of fungal enzyme action, resulting in a sorer pulp
with improved bonding and stronger paper characteristics. Reduced colour
reversion was another benefit noted with the fungus IZU-154 [154]. Some
viscosity loss, indicating limited cellulose depolymerization, has been reported
Reductionof OrganochlorineCompoundsin BleachPlant Effluents 235
as a result of fungal bleaching [153, 154, 157, 199]. However, based upon experi-
ments done with free and immobilized cultures, Kirkpatrick et al. [ 153] reported
that up to 25% of the reduction in the pulp viscosity may be due to the presence
of fungal mycelia rather than cellulose cleavage. Although fungal bleaching is
primarily an oxidative process, it appears to be more selective than oxygen
bleaching at high pH and at kappa number less than 17 because there is a better
retention of pulp viscosity.
Only a few researchers have measured the impact of fungal bleaching on
effluent quality. In a Japanese study with FCED bleaching sequence, the COD
and color in the bleach plant waste water were reduced by 50% and 80%
respectively, assuming that the filtrate from the fungal bleaching stage was sent
to a kraft chemical recovery system [1541. Whether this could occur in practice
would depend on the capacity available in the recovery furnace. The authors
suggested that higher reductions could be obtained with an FED or
FE1D1EzDz sequence, although there may be slight loss in pulp yield [1541.
Despite the emphasis on fungal bleaching as a means to reduce the use of
chlorine and the associated formation of chlorinated organics, the effect upon
chlorinated organic discharges has not been reported. As this is an important
factor in the choice of any alternative bleaching sequence, quantitative informa-
tion in this area is needed.
4 Treatment of Bleach Plant Effluents
Numerous physicochemical methods have been used for the treatment of bleach
plant effluents. These treatments include precipitation with lime [200-203],
alum and metal ions [204, 205] and synthetic polymeric coagulants [2061,
adsorption on activated carbon [207], natural clays [208-210] and polymeric
adsorbents [211, 212], membrane techniques [2131, rapid filtration on soil
[214-2161, UV-irradiation [217-2191, and oxidation using oxygen [2201, sul-
furdioxide [2211, hydrogen peroxide and sodium hypochlorite [222-224].
The problems underlying the physicochemical treatments are those asso-
ciated with cost and reliability. Coagulation and precipitation produce a vol-
uminous sludge which is very difficult to dewater. Usually, an extreme pH range
is used for optimum treatment, and the pH needs to be readjusted to neutral
before discharge. Oxidation using ozone and hydrogen peroxide are costly, and
oxidation using chlorine species generates secondary pollutants such as chlorin-
ated organics. The membrane techniques require pretreatment and a large
capital investment. Membrane fouling is also a problem with the membrane
technique.
Biotechnological methods have the potential to eliminate/reduce the prob-
lems associated with physicochemical methods, and are described in the follow-
ing section.
4.1 Biological Treatment
Biological treatment is known to be effective in reducing the BOD and simulta-

neously the toxicity of kraft mill effluents [225, 226]. Some aspects of biological
treatment have been reviewed by Boman et al. [227].
4.1.1 Using Bacteria
4.1.1.1 Aerobic Processes. Biological oxidation is the most widely used tech-
nique to remove BOD and chlorinated organics because of its effectiveness and
low cost. Rogers et al. [228] treated the bleached kraft mill effluent in a bench-
scale aerated lagoon for 29, 58 and 99 h and showed that toxicity, BOD and
resin acids were most consistently reduced during the 99 h treatment. Leach
et al. [229] reported the biodegradation of seven compounds representing the
major categories of toxicants in a laboratory batch aerated lagoon. Resin acids
(major source of acute toxicity) were readily biodegradable, but only part (less
than 30%) of the load of chlorophenolic compounds was removed
[34, 77, 229, 230]. However, Gergov et al. [230] reported that biological treat-
ment, especially the activated sludge process, removed 75-95% of chloro-
phenolics. Chlorinated neutral organic compounds (mutagenicity of the spent
liquor) are removed effectively [231]. Chloroform is stripped off by the air
during biological treatment. COD, TOC1 and high molecular weight material
are reduced to a lesser extent [44]. Eriksson and Kolar [51] have shown that the
high-molecular-mass fraction in bleach effluent cannot be degraded in an
aerated lagoon.
While the Swedish researchers found that the biological treatment is ineffec-
tive in removing TOC1, American mills reported, an average of 5 0 4 0 % TOC1
removal by an aerated lagoon or activated sludge process. Gergov et al. [230]
investigated pollutant removal efficiencies in mill scale biological treatment
systems. They found that 48-65% of AOX was removed in the activated sludge
process. The aerated lagoon was found to be less efficient than the activated
sludge. Recently, Deardorff et al. [232] reported that the efficiency of AOX
removal through biotreatment of combined bleach plant effluent increases with
increasing chlorine dioxide substitution. Biological treatment in an aerated
lagoon reduced the concentration of polychlorinated phenolic compounds by
97%. Jokela et al. [233] reported that aerobic lagoon system removed 58-60%
of the organochlorine compounds from the water phase, and the full scale
activated sludge plants removed 19-55%. Both biotreatments removed all sizes
and classes of organochlorine molecules and slightly shifted the relative size
distribution of the compounds remaining in the water phase towards the higher
molecular masses.
Graves et al. [234] examined the combined effects of oxygen delignification,
C102 substitution and biological treatment on pollutants levels in bleach plant
effluents. Biological treatment reduced COD, BOD, AOX and toxicity, but did
not reduce colour. Lafond and Ferguson [-235] reported that aerobic treatment
in activated sludge reactors removed 14-25% of AOX.
To enhance the efficiency of the biological treatment, Ek and Eriksson [236]
combined ultrafiltration and biological treatment. The result of the process was
better than the sum of the two processes. The detoxification of the effluent
during ultrafiltration was believed to be responsible for the improvement.
The mechanisms leading to the removal of chlorinated organics include
stripping of voltatiles, hydrolysis, chemical oxidation, biological oxidation, foam
separation, adsorption, and precipitation.
4.1.1.2 Anaerobic Processes. Anaerobic biological treatment can also efficiently

destroy chlorophenolic compounds, mutagenicity and acute toxicity [77, 237].
The Enso-Fenox Process was capable of removing 64-94% of the chlorophenol
load and toxicity, mutagenicity and chloroform in the bleaching effluent.
McFarlane and Greenwood [-238] removed 92% of color in kraft bleaching
effluent in a granular activated carbon anaerobic system in a 2-day hydraulic
residence time. The decolorization was decreased to 75% when the retention
time was shortened to 0.85 day. Gunnarsson et al. [239] treated 40% E stage
effluent combined with sulphite evaporator condensate in an anaerobic reactor.
Approximately 60% COD and 90% BOD reductions were achieved. The
methane yield was high, but the reduction of TOC1 was limited, which was still
better than the parallel aerobic treatment of the same effluent.
Many compounds in the bleaching effluent were reported to be toxic to
anaerobic microorganisms. These compounds include residual H202 in CTMP
bleaching effluent, chelating agents, extractives and chlorinated compounds
[240-243]. Adoption of microorganisms to the effluent containing these com-
pounds is necessary to achieve a successful treatment [-240]. Raizer-Neto et al.
[243] studied the efficiency of anaerobic treatment in reducing chlorinated
organic compounds in a fixed bed reactor. AOX removal efficiency was
primarily affected by increasing AOX concentration and was only about
50% for 50 mg/1 COD. BOD 5 removal efficiencies were affected at higher
AOX concentrations of 100 mg/1. AOX loading rate or hydraulic residence
time were found to be more important limiting factors than bleaching
effluent AOX concentration. For AOX loading rates under 40 mg/1 and an
AOX concentration as high as 130 mg/1 in the effluent feed, COD and BOD5
removal were about 80%. Fitzsimons et al. [244] reported 35-40% COD
and 42-45% AOX reductions in a 1.5-day anaerobic treatment. They found
that at least part of the higher molecular mass was dechlorinated. Since
anaerobic microorganisms are believed to be unable to degrade the high-
molecular-mass chlorolignin, the dechlorination of this could be mainly due to
physical and/or chemical action in the anaerobic process. The anaerobic de-
chlorination of chlorolignins is due to a combination of energy metabolism
growth, chemical hydrolysis and probably adsorption and/or insolubilization
[244, 245].
E K and Eriksson proposed a process based on U F and anaerobic and

aerobic biological treatments [236]. The U F was used to separate the high-
molecular-mass material, which is relatively resistant to biological degradation.
Anaerobic microorganisms were believed to be able to remove highly chlorin-
ated substances more efficiently than aerobic microorganisms. The last remain-
ing chlorine atom was removed by aerobic microorganisms. The combined
treatments typically removed 80% of the AOX, C O D and chlorinated phenolics
and completely eliminated chlorate. Lafound and Ferguson [235] reported that
anaerobic treatment in an upflow hybrid reactor removed 17-40% of AOX.
Armentate et al. [246] investigated an integrated anaerobic/aerobic process for
the biodegradation of chlorinated aromatic compounds. The sludge obtained
from the anaerobic digester of a commercial treatment plant was used to obtain
an anaerobic consortium capable of partially dechlorinating 2,4,6-trich-
iorophenol. The clarified and sterilized effluent from the same anaerobic digester
was used as the medium for the anaerobic consortium. During the anaerobic
process, 2,4,6-TCP was first dechlorinated to 2,4-dichlorophenol and then to
4-chlorophenol. The stoichiometric amount of 4-CP was recovered. Similar
results were obtained when the anaerobic microorganisms were immobilized.
After immobilization, the consortium was able to dechlorinate 150 laM of
2,4,6-TCP in four days. Pseudomonas 91athei and an indigenous culture obtained
from same sludge used to produce the anaerobic enrichment culture were shown
to be able to degrade the 4-CP produced from the anaerobic dechlorination of
2,4,6-TCP.
Strehler and Welander [247] investigated the removal of AOX and C O D
from bleached kraft mill effluent in laboratory and pilot scale aerobic suspended
carrier reactors and abiotic thermoalkaline reactors. At pH 7.0, 37 ~ and HRTs
longer than 3.5 hours, a maximum C O D removal of 55% was achieved in the
suspended carrier process. The C O D conversion rate at the minimum H R T was
2.6 kg C O D m 3 d-1. The suspended carrier treatment was operated success-
fully at pH 9.0 and 45 ~ and at pH 7.0 and 50 ~ giving > 50% C O D removal
with an H R T of four hours. The AOX removal achieved at p H 9.0 and 45 ~
(50%) was significantly higher than that at pH 7.0 and 37 ~ (39%) because of an
increased abiotic dechlorination at the higher p H and temperature levels. These
researchers further studied sequential thermoalkaline and biological treatment
on a pilot scale. Thermoalkaline treatment at pH 10.0, 54 ~ and an H R T of two
hours followed by biological treatment at pH 8.0, 35 ~ and an H R T of four
hours removed almost 80% of the AOX and 50% of the C O D from the kraft
mill effluents.
4.1.2 Using Fungi
The reason why bacteria show low efficiency in removing COD, TOC1 and
high-molecular-mass chlorolignins from bleaching effluent is that bacteria de-
grade the substrate by an intracellular enzyme system. The substrate must be
Reduction of Organochlorine Compoundsin Bleach Plant Effluents 239
able to pass the bacterial cell membrane in order to be degraded. In the light of
this, the inefficiency of bacteria in degrading high-molecular-mass chlorolignins
is understandable. Since most TOC1 and COD is due to the high-molecular-
mass chlorolignins, low COD and TOC1 removal efflciencies are also to be
expected.
An approach to degrading high molecular weight chlorolignins is to use
white-rot fungi, the only known microorganisms to efficiently degrade lignin.
They generate a lignolytic activity to degrade lignin in a so-called secondary
metabolism stage when one of several nutrients, nitrogen, phosphorous or
carbon is depleted (unfavorable natural conditions). White-rot fungi excrete an
operating system including extracellular enzymes which can degrade high-
molecular-mass chlorolignins effectively. So far, the widest applications of the
white-rot fungi in waste-water purification have been concentrated on the
decolorization of bleaching or pulp mill effluents [248-250].
The reaction mechanism by which white-rot organisms degrade lignin is in
four steps. Ligninase enzymes catalyze the first two steps. This is followed by an
aromatic hydroxylation step that produces catechol structures in the resulting
fragments [251, 252]. The fourth step is an oxidative ring cleavage catalyzed by
a dioxygenase enzyme. The products of this reaction are converted by the
organism into CO2 and water. Chlorinated aromatics are dealt with by the
organism in a like manner by converting them into catechol structures whose
rings can be cleaved via the same dioxygenase enzyme, the products being CO2,
water and inorganic chlorides. Many other organisms can grow directly on
chlorinated organics, degrading them via the same or similar dioxygenase
pathways or in some cases monooxygenase pathways.
The best known white-rot fungus is Phanerochaete chrysosporium. This
fungus is known to secrete a family of enzymes which degrade both lignin and
modified lignin/-249]. Both high- and low-molecular-mass chlorolignins gener-
ated during the pulp bleaching are significantly degraded [253, 254]. The
fungus reduces the COD by degrading the chlorolignins to CO2 and chloride,
decolorizes the bleaching effluent by destroying both the color bodies and
chromophoric structure, and removes TOC1 by converting it to inorganic
chloride [253-255]. Most of the low-molecular-weight chlorophenolics disap-
pear after 1 day's fungal treatment [256, 257]. This particular fungus is also
known to be able to degrade refractory compounds such as TNT, PCB and
Lindane, DDT, chlorinated dioxin and other difficult biodegradable com-
pounds [258-260].
Decompositions of lignin to CO2 by the lignin-decomposing fungus
P. chrysosporium requires a growth substrate such as cellulose or glucose.
Growth on lignin as a sole carbon source is negligible. In addition to its
requirement for a co-substrate, the ligninolytic system (a) is produced even in the
absence of lignin, (b) is expressed only during secondary metabolism, (c) is
triggered by carbon, sulfur, or nitrogen limitation, (d) is strongly repressed by
glutamate and certain other amino acids, (e) is sensitive to the balance of trace
metals supplied, (f) has a relatively narrow pH optimum (4 5), and (g) is
markedly affected by oxygen concentration [-261-265]. The fungal decoloriz-

ation, like lignin metabolism, is a secondary metabolic event and is probably
attributable to lignin metabolism since lignin and its degradation products are
the main color source in E1 effluent. The optimum conditions favoring fungal
growth are quite different from those favoring decolorization. The pH range for
optimum growth is 4.3~4.8 and decolorization is greatly retarded below pH 4.0
or above 5.0 because of poor growth [261]. The optimum temperature for the
growth of fungus is 40 ~ whereas decolorization is not limited to the same
narrow range of temperature, but takes place with little decrease in rate at
temperatures as low as 25 ~ [261]. The fungal decolorization requires oxygen
and a co-substrate, but, unlike fungal growth, the addition of a nitrogen source
is not necessary. Eaton et al. [266] have outlined a process based on laboratory
and bench scale experiments for decolorization by P. chrysosporium. The fungi
require a growth substrate for decolorization just as they do for lignin degrada-
tion. Investigations have demonstrated that the cellulose-rich primary sludge
serves this purpose well. The fungus requires a fixed surface for pregrowth
without agitation in shallow medium for efficient color reduction. Fixed film
reactors of the rotating biological contactor (RBC) design, which are commer-
cially available and are presently used in waste treatment have been proved to
be effective. Color removal is greatly stimulated by an oxygen-enriched atmo-
sphere. The RBC provides good aeration and can be enclosed for the addition of
supplementary oxygen. A growth stage is necessary before decolorization be-
gins. During this stage, nutrient nitrogen is depleted, and the fungus becomes
ligninolytic and able to decolorize. Based on the above considerations, the
process shown in Fig. 3, which has been termed the FPL/NCSU/MyCoR
process [267], can be outlined. This mycelial color removal process resulted
from the cooperative research between the US Forest Products Laboratory and
North Carolina State University. A fixed film MyCoR reactor is charged with
growth nutrients, which can include primary sludge as the carbon source, and is
inoculated with a suitable fungus. Depending on the mill, the sludge will provide
Influent
uent
Fig. 3. Rotatingbiologicalcontactorfor fungaldecolorizationof pulp mill effluent

some of the required mineral nutrients and trace elements as well as carbon.
Nitrogen-rich secondary sludge can also be used to supply the nitrogen required
for growth. After the mycelium has grown over the reactor surface, it depletes
the available nitrogen and becomes ligninolytic (pregrowth stage 2 to 4 d). The
reactor is then ready for use. Operation for over 60 days has been achieved in
bench reactors in a batch mode. The process converts 70% of the organic
chlorides to inorganic chloride in 48 h while decolorizing the effluent and
reducing both COD and BOD by about half. Huynh et al. [268] used the
MyCoR process for the treatment of the chlorinated low-molecular-mass phen-
ols of the E1 effluent. Their results showed that most of the chlorinated phenols
and other low-molecular-mass components of the effluent were removed during
fungal treatment. Pellinen et al. [269] have reported that the MyCoR process
can be considerably improved in terms of COD removal by simply using less
glucose as the carbon source for the fungi, P. chrysosporium. However, the
dechlorination was reported to be faster at high glucose concentration. The
kinetics of decolorization of E1 effluent with P. chrysosporium in an RBC under
improved conditions have been studied by Yin et al. [270]. The kinetic model
developed for 1 and 2-d retention times showed a characteristic pattern. The
overall decolorization process can be divided into three stages, viz. a rapid color
reduction in the Ist hour of contact between the effluent and the fungus followed
by a zero order reaction and then a 1st order reaction. The color removal rate on
the second day of the 2-d batch treatment was less than that on the first day. The
decolorization in a continuous flow reactor achieved approximately the same
daily color removal rate, but the fungus had a longer working life than when in
the batch reactor, thereby removing more color over the fungal lifetime. Pellinen
et al. [271] studied dechlorination of high-molecular-mass chlorolignin in first
extraction stage effluent with white-rot fungus P. chrysosporium immobilized on
an RBC. The total organic chlorine content of chlorolignin decreased almost by
50% during one day of treatment. Correlation studies suggested that dechlorina-
tion, decolorization and degradation of chlorolignin (as COD decrease) are
metabolically connected, although these processes have different rates. Size exclu-
sion chromatography showed that polymerization took place in the early stage of
the treatment. Low-molecular-mass degradation products were not observed.
A sequential biological treatment using this fungus and these bacteria was
studied to decrease TOC1, color and COD in conventional softwood kraft pulp
bleaching effluent by Yin et al. [272]. In six variations of the white-rot fun-
gus/bacteria system studied (Table 8), only the degree of fungal treatment was
varied. In three of the six variations, ultrafiltration was also used to concentrate
high-molecular-mass chlorolignins and to reduce effluent volume (and thus cost)
prior to fungal treatment. The best sequence, using ultrafiltration/white-rot
fungus/bacteria, removed 71% TOC1, 50% COD and 65% color in the effluent
(Table 8). Fungal treatment enhances the ability of bacteria to degrade and
dechlorinate chlorinated organics in the effluent. The fungus is able to remove
organic chlorine from chlorolignins and to attack the high- and low-molecular-
mass chlorolignins.
~ 2 2 2 2 2
e~
ii! ---- ='" ~
,--M
O
O
i
!
!
.r,
=~
i
! II II ~ - 2 , =
0
Guo et al. [257] investigated degradation of model compounds - chloro-

phenols (pentachlorophenols, 2,4,6-trichlorophenol, and 2,4-dichlorophenol)
and chloroguaiacols (4,5-dichloroguaiacol and 4,5,6-trichloroguaiacol) in pure
water solution by fungal treatment using an RBC. They found that at concentra-
tion of 30 mg/1, 80-85% of chlorophenols and chloroguaiacols could be de-
graded after 3-4 h of treatment. Meanwhile, some of these compounds were
methylated to chloroanisoles and chloroveratrols.
Messner et al. [273] developed a similar process, i.e. the MyCoPOR. In this
process, porous carrier material is inoculated with spores of Phanerochaete
chrysosporium. Shaking-flask tests with and without foam as porus carrier
material showed that the decolorization of an E1 effluent from a sulfite plant was
improved from 50% without foam as supporting material to 70-80% with foam.
Decolorization was accompanied by about 70% AOX reduction when using
foam. If nutrients were replaced and effluent removed daily, the active lifetime of
the mycelium was 6-8 weeks, depending on the fungal strain. A trickling filter
system was found to be the most effective for the degradation of effluents. With
daily replacement of the medium/effluent solution, the decolorization rate of the
E1 effluent was 70% when aerated by air, and AOX reduction was 60%. The
fastest degradation of the effluent took place during the first 3-6 hours, when
about 50% of the color and AOX were reduced.
Another white-rot fungus, which has shown good performance, is Coriolus
versicolor. It requires a growth substrate such as cellulose or glucose for the
decomposition of lignin to carbon dioxide [274]. The culture conditions favor-
ing lignin degradation are similar to those favoring fungal decolorization.
Livernoche et al. [274] showed that C. versicolor in liquid culture removed over
60% of the color of the combined bleach kraft effluent within 6 d in the presence
of sucrose. Decolorization of effluent was more efficient when the concentration
of sucrose (growth substrate) and inoculum was high. Treatment of the same
effluent with this fungus, immobilized in beads of calcium alginate gel, resulted
in 80% decolorization after 3 d in the presence of sucrose. The pH of the effluent
during decolorization decreased from 5.7 to 3.6, probably due to formation of
organic acids resulting from the metabolism of the immobilized fungus. How-
ever, decolorization was not due to lowering of pH. The decolorization process
affected not only the dissolved chromophores but also the suspended solids. The
solids after centrifugation of the zero time samples were dark brown, while the
solids after 4-d incubation were light brown. The beads with the immobilized
mycelium remained light-coloured throughout the experiments with no indica-
tion of accumulation of the effluent chromophores. Decolorization was achieved
more rapidly at pH 5.0 than at pH 7.0. Recycled beads could remove color
efficiently and repeatedly in the presence of air but not under anaerobic condi-
tions.
Biological reactors of the airlift type using calcium alginate beads to immo-
bilize the fungus C. versicolor have been used to study the continuous decoloriz-
ation of kraft mill effluents [275]. The effluent used contained only sucrose and
no other nutrient source. An empirical kinetic model was proposed to describe
the decolorization process caused by this fungus, but it did not shed any light on
the chemical mechanism involved in the decolorization.
Direct use of suspended mycelium of the fungus, C. versicolor, may not be
feasible because of the problem of viscosity, oxygen transfer and recycling of the
fungus. The fungus was, therefore, grown in the form of pellets, thus eliminating
the problems with biomass recycling and making it possible to use a larger
amount [276]. Rate of decolorization with fungal pellets was almost ten times as
high in batch culture as in continuous culture under similar conditions. The
capacity for decolorization decreased markedly with increase of lignin loading
[-276].
Bergbauer and Kraepelin [277] showed that C. versicolor efficiently de-
graded chlorolignins from bleaching effluent. More than 50% of the chlorolig-
nins were degraded in a 9-d incubation period, resulting in a 39% reduction in
AOX and an 84% decrease in effluent color. In a 3-1 laboratory fermenter with
0.8% glucose and 12 m M ammonium sulphate, about 88% color reduction was
achieved within 3 d. Simultaneously, the concentration of AOX dropped from
initially 40 mg/1 to 21.9 mg/1, a 45% reduction within 2 d. With malt extract
instead of glucose, reduction of color unit as well as AOX values were nearly the
same [278].
Bajpai et al. [279] used pellets of T. versicolor strain B-7 for decolorization
of E1 effluent. The mycelial pellets oxidized the chromophores of the effluent in
the presence of any of the cosubstrates sucrose, glucose, starch, ethanol, car-
boxymethylcellulose, microcrystalline cellulose, pulp and malt extract. The
highest decolorization was obtained in the case of glucose. Optimum pH and
temperature were 4.5-5.5 and 30 ~ respectively. In the batch reactor with an
effluent of 7000 color units per litre, the maximum color reduction of 93% was
obtained in 48 h with a C O D reduction of 35%, whereas, in a continuous
reactor, the same level of color and C O D reduction was obtained in 38 h
residence time. No loss in decolorization ability of mycelial pellets was obtained
when the reactor was operated continuously for more than 30 d. They also used
T. versicolor for decolorization of effluent from a pulp mill utilizing agriresidues
[280]. With an effluent of 18 500 color units, a maximum color reduction of 92 %
with a C O D reduction of 69% was obtained.
Royer et al. [281] described the use of the pellets of C. versicolor to
decolorize ultrafiltered kraft liquor in nonsterile conditions with a negligible loss
of activity. The rate of decolorization was observed to be linearly related to the
liquor concentration and was lower than that obtained in the M y C o R process.
This could be due to the lower temperature (22 ~ used in this work and to the
use of pellets with relatively large diameters which could limit the microbial
activity as compared to the free mycelium used in the M y C o R process. An
effective decolorization of effluent having 400-500 color units/1 can be obtained
in presence of a simple carbon source such as glucose. In the repeated batch
culture, the pellets exhibited a loss of activity dependent on the initial color
concentration. The production of the extra cellular enzyme, laccase, was fol-
lowed but could not be used as an indicator of the ligninolytic activity.
Archibald et al. [282] studied the fungal requirements for optimal growth
and decolorization and the mechanism of chromophore degradation by
C. versicolor. Simple carbohydrates were essential for effective decolorization,
and a medium composed of inexpensive industrial by-products provided excel-
lent growth and decolorization. The regulation or control of E1 effluent decolor-
ization appears to differ substantially from that of lignin peroxidase production
and the induction of ligninolytic activity. In particular, modulation of decoloriz-
ation by trace metals, nitrogen and carbon limitation, culture age and veratryl
alcohol was not observed.
Marton et al. [283] reported that C. versicolor reduced the characteristic
dark brown color of diluted alkaline lignin solutions. They found, however,
about half of the color and one fourth of the aromatically absorbing substances
were recovered from the fungal cells by alkaline extraction. Therefore, it was
concluded that adsorption played a major role in lignin degradation. Decoloriz-
ation also proceeded anaerobically but to a lesser extent.
Treatment of E1 stage eff with ozone and the fungus C. versicolor has
also been tried [284]. Both ozone treatment and biological treatment effectively
destroyed effluent chromophores, but the fungal process resulted in greater
degradation as expressed by COD removal. Monoaromatic chlorophenolics
and toxicity were removed partially by ozone and completely by C. versicolor.
Molecular weight distributions showed roughly equal degradation of all sizes of
molecules in both treatments. The combination of a brief ozone treatment with
a subsequent fungal treatment revealed a synergism between the two decoloriz-
ation mechanisms on E1 stage effluent. Effluent was pretreated with
ozone (ll0-160mg/1) or C. versicolor (24h with 2-5g/1 wet weight
fungal biomass). The pretreatment was followed by a 5-d incubation with C.
versicolor. It was noted that partial color removal by ozone pretreat-
ment allowed more effective removal by the fungus than that by fungal pre-
treatment. After 20h, 46-53% decolorization was observed for ozone-
pretreated effluents, compared to 29% for fungal treatment alone. The contribu-
tion of ozone seemed to be most important in the first 24 h following the
pretreatment. Ozone pretreatment also produced a small improvement in the
bioavailability of effluent organics to the fungus. A partial replacement of
chlorine by ozone in the bleach plant (the addition of an ozone bleaching step)
or a brief ozone pretreatment of E1 stage effluent should considerably reduce the
low-molecular-mass toxic chlorophenolics. In addition, the use of ozone should
also improve decolorization by subsequent fungal and possibly bacterial treat-
ments.
Another white-rot fungus, Schizophyllum commune, has also been found to
decolorize and degrade lignin in pulp and paper mill effluent [-285]. The fungus
was able to degrade lignin in the presence of an easily metabolizable carbon
source. The addition of carbon and nitrogen not only improved the decolorizing
efficiency of the fungus but also resulted in reduction of the BOD and COD of
the effluent. Sucrose was found to be the best carbon source for the breakdown
of lignin. A 2 d incubation period was sufficient for lignin degradation by
S. commune. The efficiency of treatment of effluent with this fungus was highest
at pH 4 5 and was further improved by intermittent aeration. Under
optimum conditions, S. commune reduced the colour of the effluent by 90%
and also reduced BOD and COD by 70% and 72% during a 2 d incu-
bation.
A white-rot fungus, Tinctoporia borbonica, has been reported to decolorize
the kraft waste liquor to a light yellow color [286]. About 99% colour reduction
was achieved after 4 d of cultivation. Measurement of the culture filtrate by
ultraviolet spectroscopy showed that the chlorine-oxylignin content also de-
creased with time, and measurement of the culture filtrate plus mycelial extract
after 14 d of cultivation showed the total removal of the chlorine-oxylignin
content.
Addition of a carbon and nitrogen source was found to improve decoloriz-
ation of pulp and paper mill waste water by the fungus Aspergillus niger, leaving
19% of the original color, and reduced about 43% BOD and 41% COD after
2 d of incubation [287]. Tono et al. [288] reported that Aspergillus sp and
Penicillium sp achieved 90% decolorization in one week's treatment at 30 ~ and
at pH 6.8. Later, Milstein et al. [289] reported that these microorganisms
removed appreciable levels of chlorophenols as well as chloroorganics from the
bleach effluent.
Prasad and Joyce [290] used Trichoderma sp., one of the fungi imperfectii, to
decolorize the hardwood E1 stage effluent. Glucose was found to be the most
effective carbohydrate utilized by the fungus, as it stimulated substantial color
reduction without any increase in COD. Addition of nitrogen did not stimulate
the decolorization process, indicating that it is not a rate-limiting factor. The
optimum pH for decolorization and growth was 4.0. Under optimal conditions,
total colour and COD decreased by almost 85% and 25% respectively after
cultivation for 3 d.
Since the majority of TOC1 and color is due to higher molecular
weight chlorolignins, future research should concentrate on the fate of high-
molecular-mass chlorolignins in biological treatment or in the natural environ-
ment. Since bacteria significantly degrade only those chloroorganics of molecu-
lar mass less than 800-1000, research is needed to decrease the chlorolignin
molecular mass or to remove high molecular mass chlorolignins before biolo-
gical treatment is applied, in order to enhance the biotreatability of bleaching
effluent.
Three approaches to decreasing the molecular weight of chlorolignin should
be studied: (1) a modified bleaching process such as oxygen prebleaching and
a high level of chlorine dioxide substitution, (2) a two-stage biological treatment
such as white-rot fungus followed by biological treatment, and (3) physicochemi-
cal treatment followed by biological treatment.
Ultrafiltration can separate high molecular weight material, and biological
treatability of permeate can be enhanced. Another method is precipitation,
which preferentially removes high-molecular-mass chlorolignins. The treated
effluent should be more amenable to biological treatment.
4.2 Enzymatic Treatment
Some enzymes also seem to have the potential to remove color and AOX from
pulp and paper mill effluents. Peroxidase, laccase etc. are the most important of
these. The use of microbial or enzyme-based treatment offers some distinct
advantages over physical and chemical AOX precipitation methods, in that only
catalytic and not stoichiometric amounts of the reagents are needed, and the low
organic concentrations and large volumes typical of bleaching effluents are,
therefore, less of a problem. Also, both complete microbial systems and isolated
enzymes such as peroxidases and laccases have been shown to reduce the acute
toxicity by polymer and thereby rendering less soluble many of the low-
molecular-mass nonchlorinated and polychlorinated phenolics [291 295]. It
has been reported that lignin-type peroxidases secreted by white-rot fungi are
involved in the degradation of a whole range of organic pollutants including
PCB, D D T and Indane, chlorinated anilines, polychlorinated phenolics includ-
ing P C P and chlorolignins, and many more [296-300]. Lyr [301] reported that
laccase of T. versicolor partially dechlorinates PCP, and Hammel and Tardone
[302] reported that peroxidase from P. chrysosporium can partially dechlorinate
P C P and 2,4,6-trichlorophenol. Arcand and Archibald [303] carried out a sys-
tematic study on direct dechlorination of chlorophenolic compounds in pulp
and paper mill effluents by laccase from T. versicolor. It was found that most of
the laccase secreted by T. versicolor could partially dechlorinate a variety of
chlorophenolics. The rate and extent of C1- release were found to be substan-
tially affected by substrate and enzyme concentration and the presence of
multiple laccase substrates. The dechlorination was found to be accompanied by
extensive polymerization of the substrate. Table 9 shows a comparison of the
removal of chlorophenolics from a mixture in E1 effluent by crude laccase with
Table 9. Removalof chlorophenoliccompoundsfrom a mixture in high-molecular-mass

E1 effluent by crude laccase in 30 mina and T. versicolor myceliumin 3 h
Reduction (%)
Chlorophenolic Concentration
Compounds (gM) Laccase Mycelium
2-Chlorophenol 137 35.5 _+1.4 61.5 2.7
3-Chlorophenol 18 51.9 + 4.3 2.0 0.1
4-Chlorophenol 31 48.1 2.9 41.6 1.3
2,4-Dichlorophenol 21 45.0 2.3 64.9 1.8
2,4,6-Trichlorophenol 26 72.4 _+5.2 83.8 6.2
2,3,4,6-Tetrachlorophenol 18 40.7 8.2 71.2 2.7
Pentachlorophenol 24 34.0 8.9 82.1 3.2
4,5-Dichlorognaiacol 52 100 96.8 + 6.1
4,5,6-Trichloroguaiacol 32 100 100
Tetrachloroguaiacol 17 76.5 21.5 95.0 7.6
aReactions were conducted in 20 ml of E1 effluent containing 20 mM D-glucose,pH 4.6,
temp. 25 ~ shaken at 200 rpm in triplicate using either 0.1 U/ml crude T. versicolor
laccase or 0.5 g wet weight (25 mgd.w.) washed T. versicolor mycelium
Based on data from Ref. [3033
that by T. versicolor mycelium, and Table 10 shows the effects of crude laccase
on various chlorophenolics. Arcand and Archibald [303] also studied the
effects of horseradish and P. chrysosporium peroxidases on the mixture of five
chlorophenolics. Both peroxidase enzymes were found to degrade the majority
of all substrates except PCP, whereas the P. chrysosporium peroxidases
was superior to both horseradish peroxidase and laccase in degrading PCP, it
was inferior to horseradish peroxidase in degrading the other four phenolics
(Table 11).
Paice and Jurasek studied the ability of horseradish peroxidase to catalyze
color removal from bleach plant effluents [304]. The color removal from
effluents at neutral pH by low levels of hydrogen peroxide was enhanced by the
addition of peroxidase. No precipitation occurred during the decolorization
process. The catalysis with peroxidase (20 mg/1) was observed over a wide range
of peroxide concentrations (0.1-800 mM) but the largest effect was between
1 mM and 100 mM. The pH optimum for catalysis was around 5.0. Compared
with mycelial color removal by C. versicolor, the rate of color removal by
peroxide plus peroxidase was initially faster (for the first 4 h), but the extent of
color removal after 45 h was higher with the fungal treatment. Further addition
of peroxidase to the enzyme-treated effluents did not produce additional cataly-
sis. Thus, the peroxide/peroxidase system did not fully represent the metabolic
route used by the fungus. One working hypothesis has been proposed to explain
the behaviour of enzymes in the decolorization process [304]. Glucose is used by
the cell to produce peroxidase. One of the extracellular enzymes often found in
white-rot fungi is peroxidase. It seems that this enzyme oxidizes the chromo-
phores and so removes the color from bleaching waste water.
Forss et al. [305] have shown that aeration in the presence oflaccase leads to
a reduction by over 90% in the amount of phenolic compounds in waste water.
They have shown that the amount of chlorophenols is also reduced by 75-99%,
depending on the group of chlorophenols (Table 12). The total reduction was
80%. The sample was aerated in the presence of laccase for 1 h and was
flocculated with aluminium sulfate. Since polymerization by laccase can be
avoided and the degradation of the substrate enhanced by the presence of
appropriate reductant, it may be possible to substantially increase the rate and
extent of laccase-driven dechlorination.
Call (of Lignozym, Germany) has patented a process for decolorization and
decontamination of waste water from pulp and paper mill effluents using
enzyme [306]. In this process, laccase enzyme from C. versicolor is used along
with phenol or nonphenolic aromatic compounds with continuous passing of
oxidation materials (H202, 02 or oxygenated air). Almost complete polymeriz-
ation of the lignin contained in the waste water is achieved, which is 20-50%
above the values attainable with the addition of laccase alone. About 70-90%
lignin is developed into insoluble form, which is removed by flocculation and
filtration.
Ferrer et al. reported that immobilized lignin peroxidase decolorized
kraft effluent [307]. Novel lignin peroxidases called Pulpases produced by
I I I I I
I I I I I '~
2 ,_
+i+i+l+l+l
+i +1 +i +1
~ cM
M ~5~
+1 +1 +1 ~1 ~1
B
"S
b
0 ~0 0 0 ' ~ . ~ ;> ~ ~'~
o 0 0
9~ ~ ~ ~ . ~ = o ~
6
250 p. Bajpai and P.K. Bajpai
.2
"0~ "~
% J
=
8 +1+1 +1+1
vv c:5
o
e.! ,,4 ~
o
o
o o,--- ~ o
O0 r--
o
e~
b
de.!
8
+1+1+1+1+1+1
de.,
o --a
o0_~ U
V t': ~ N , ~
~ . j
m
o " ~ ~ ~ ~..
,.-8 ~ o o ~,
o =.~ = .~ ~ "-6 O.=oO
o No ~
0
l
,It
Table 12. Concentration of chlorinated phenols in bleach plant effluent from a softwood
kraft mill before and after aeration in the presence of laccase and flocculation with alum
Phenolic compound Concentration (rag/l) Reduction (%)
Initial After 1 h aeration
Chlorinated phenols 141 9 93.6

Chlorinated vanillins 813 176 78.3
Chlorinated guaiacols 516 30 94.2
Chlorinated catechols 251 19 92.4
Total 1721 234 86
Based on data from ref. [305]
P. chrysosporium m u t a n t strain SC 26 a n d d e s c r i b e d in 2 p a t e n t s assigned to the

Repligen C o r p o r a t i o n [308, 309] have been c l a i m e d to d e c o l o r i z e b l e a c h i n g
effluents.
5 Conclusions
M a n y process c h a n g e s have been i m p l e m e n t e d o r are being c o n s i d e r e d to

reduce the f o r m a t i o n of A O X a n d c h l o r i n a t e d d i o x i n s from c h e m i c a l p u l p
b l e a c h i n g o p e r a t i o n s . T h e r e are also possibilities for t r e a t m e n t of effluents with
m i c r o o r g a n i s m s a n d enzymes to r e m o v e o r d e c h l o r i n a t e o r g a n i c material. E a c h
o p t i o n discussed has i n h e r e n t a d v a n t a g e s a n d d i s a d v a n t a g e s with r e g a r d to
c a p i t a l costs, o p e r a t i n g costs, ease of retrofit, f a b r i c a t i o n a n d i n s t a l l a t i o n time.
I m p a c t on o t h e r mill unit o p e r a t i o n s is also c o n s i d e r e d in c h o o s i n g the best
options. M a n y factors h a v e to be c o n s i d e r e d in c h o o s i n g an effective a n d
e c o n o m i c a l b l e a c h i n g / t r e a t m e n t process t h a t meets all the e n v i r o n m e n t a l guide-
lines. It a p p e a r s t h a t the o n l y l o n g - t e r m s o l u t i o n will p r o b a b l y be to d e v e l o p the
t e c h n o l o g y which will allow mills to o p e r a t e with zero effluent.
Acknowledgements We dedicate this article to Shri L.M. Thapar (Chairman, Thapar Group of
Industries) on his 65th Birthday, thanking him for his never-failing encouragement. We also thank
Dr. M.P. Kapoor, Director, Thapar Corporate R&D Centre, for his encouragement, Dr. M.B.
Jauhari, General Manager (R&D), Ballarpur Industries Limited, for helpful discussions and Shri S.S.
Gill for excellent typing of the manuscript.
6 References
1. Rennel J (1995) Nordic Pulp and Paper Res J 1:34

2. Casey JP (1980) Pulp and Paper: Chemical and Chemical Technol, 3rd edn, vol 1 Wiley, New
York, p 681
3. Lindberg S (1974) Chem Abs 80 1,407, 404m

4. Voss R, Wearing J T , Wong A (1981) Pulp and Paper Can 82(2): T65
5. Bjorseth A, Carlberg GE, Gjos N, Moller M, Tveten G (1981) In: Keith LH (ed) Advances in
the Identification and Analysis of Organic Pollutants in Water, vol 2. Ann Arbor Science, Ann
Arbor, p 1115
6. Salkinoja-Salonen M, Saxelin ML, Pere J, Jaakkola T, Saarikoski J, Hakulinen R, Koistinen
O (1981) In: Keith LH (ed) Advances in the Identification and Analysis of Organic Pollutants
in Water, vol 2, Ann Arbor Science, Ann Arbor, p 1131
7. Berry RM, Luthe CE, Voss RH, Wrist PE, Axegard P, Gellerstedt G, Lindblad PO, P6pke
I (1993) In: Jameel H (ed) Bleaching, vol 2, Tappi Press, Atlanta, p 759
8. Gergov M, Priha M, Talka E, Valltila O, Kangas A, Kukkonen K (1988) Tappi J 71(12): 175
9. Lindstr6m K, Mohamed M (1988) Nordic Pulp Paper Res J 3:26
10. Kringstad KP, Lindstrbm K (1984) Environ Sci Technol 18: 236A
11. Mckague AB, Jarl M, Kringstad KP (1989) Proc of the Fifth Internat Symp of Wood and
Pulping Chemistry. Myrtle Beach, SC
12. Voss RH (1983) Environ Sci Technol 17:530
13. Voss RH, Wearing JT, Wong A (1980) In: Chemical Congress in the North America Continent,
Las Vegas
14. Axegard P (1984) Pulping Conf Proc, Tappi, Atlanta, p 353
15. Annergren G, Kringstad KP, Lehtinen KJ (1986) In: Proc of EuCePa Symp Environmental
Protection in the 90's, Helsinki, Finland, May 19-22, p 40
16. Alfthan CJ, Norrodtrom H, Akerlund G (1976) Svensk Papperstidn 79:180
17. Voss RH, Wearing JT, Wong A (1980) Pulp and Paper Can 81(12): T367
18. Crawford RJ, Stryker MN, Jett SW, Carpenter WL, Fisher RP, Jain AK (1987) Tappi J 70(11): 123
19. Chan RW, McDonald JR (1983) In: 1983 Technical Section, Annual Meeting Preprints, CPPA,
Montreal, p 239
20. Kringstad KP, Mckague AB (1988) In: Proc of 1988 lnternat Pulp Bleaching Conf, Orlando,
FL, p 63
21. Sodergren A, et al. (1987) Summary of Results from the Swedish Project Environment/cellulase.
Symp Reprints, The second IAWPRC Symp on Forest Wastewaters Biological Treatment
and Environmental Effects of Pulp and Paper Industry Wastewaters, Tampere, Finland
22. Suntio L, Shiu WY, Mackay DA (1988) Chemosphere 17(7): 1249
23. Dence CV, Wang C J, Durkin PR (1980) Toxicity Reduction through Chemical and Biological
Modification of Spent Pulp Bleaching Liquors, EPA Report 600-2-80-039, US EPA, Athens
24. Walden CC, Howard TE, Tappi J (1977) 60(1): 122
25. Ander P, Eriksson KE, Kolar MC, Kringstad K (1977) Svensk Papperstidn. 80(14): 454
26. Hutchins FE (1979) Toxicity of Pulp and Paper Mill Effluent. A Literature Review, EPA-
600/3-79-013, US Environmental Protection Agency
27. Eriksson KE, Kolar MC, Kringstad KP (1979) Svensk Paperstidn. 82(4): 95
28. Priha M, Talka E (1986) In: Proc of EuCePa Symp Environmental Protection in the 90's,
Helsinki, Finland, May 19-22, p 50
29. Kringstad KP, Lindstrom K (1988) In: Proc of 1988 Internat Pulp Bleaching Conf, Orlando,
FL, p 5, Addendum
30. Carlberg GE, Kringstad A, Martinsen K, Nashang O (1986) In: Proc of EuCePa Symp
Environmental Protection in 90's, Helsinki, Finland, p 55
31. Bjorseth A, Carlberg GE, Moller M (1979) Sci Total Environ 11(2): 197
32. Leach JM, Muller JC, Waden CC (1977) Trans Tech Sect, CPPA 3(4): TR126
33. Tolan JS, Canovas RV (1992) Pulp and Paper Can 93(5): 39
34. Holmbom B, Lehtinen KJ (1980) PaperiPuu, 62(11): 673
35. Tsai TY, Renard JJ, Phillips RB (1994) Tappi J 77(8): 149
36. McCubbin N (1989) Pulp and Paper Can 90(11): 17
37. McCubbin N, Sprague JB, Bonsor N (1990) Pulp and Paper Can 91(3): 112
38. Chem and Eng News, 6-7, (1994)
39. Hileman B, Long JR, Kirschner EM (1994) Chem and Eng News, 12-26
40. Kachi Set al. (1979) Paper presented at the 1979 CPPA/TS Environ Conf, Victoria, B.C.
41. Lindstrom K, Nordin J (1976) J of Chromatogr 128:13
42. Davis JC (1973) J Fish Res Rd Can 30:369
43. SSVL Project (1982) Environmentally Harmonized Production of Bleached Pulp Final Report,
Stockholm
44. SSVL-85 (1985) Project 4 Production of Bleached Pulp Final Report, Stockholm
45. Larsson A, Andersson T, Forlin L, Hardig J (1987) Symp Preprints, The Second IAWPRC
Symp on Forest Wastewaters Biological Treatment and Environmental Effects of Pulp and
Paper Industry Wastewaters, Tampere, Finland
46. Carlberg GE, Kringstad A, Martinsen K, Nashaug O (1987) Paperi ja Puu 69:337
47. Annergren G, Carlsson G, Norrby M (1987) In: Proc of the Second IAWPRC Symp on Forest
Industry Wastewaters, Tampere, Finland
48. "The Action Plan for Marine Pollution" (1987) Swedish National Environmental Protection
Board, Solna, Sweden
49. Hall TJ, et al. (1985) Effects of biologically treated bleached kraft mill effluent on cold water
stream productivity in experimental channels Fourth Progress Report. Technical Bulletin
No. 474, National Council of the Paper Industry for Air and Stream Improvement, New York
50. Sagfors PE, Starck B (1987) In: Proc of the 2nd IAWPRC Symp on forest Industry Waste-
waters, June 9 12, Tampere, Finland
51. Eriksson KE, Kolar MC (1985) Environ Sci Technol 19:1086
52. Eriksson KE, Kolar MC, Ljungquist PO, Kringstad KP (1985) Environ Sci Technol 19:1219
53. Hardell HL, de Sousa F (1977) Svensk Papperstidn. 80(4): 110
54. Germgard U (1988) In: 1988 Pulping Conf Proc, Tappi Press, Atlanta, p 315
55. Andrews EK, Chang HM, Jameel H (1991) In: Proc of the Internat Pulp Bleaching Conf,
Stockholm, Sweden, Vol 3, p 67
56. Boman R, Dahl M, LindstriSm LA, Norden S (1991) In: Proc of the Internat Pulp Bleaching
Conf, Vol 3, p 35
57. Mocas TS, Jiang EJ, Becker EX, Greenwood BF (1991) In: Proc of the Internat Pulp Bleaching
Conf, Stockholm, Sweden, Vol 3, p 70
58. Johnson AP (May 1994) Appita 47(3): 243
59. Perala J, Germgard U (1993) Presented at the 1993 CPPA Annual Meeting, Montreal, Book A,
p 281
60. Sjobolm K, Hartler N, Mjoberg J, Sjodin L (1983) Tappi J 66(9): 97
61. Tikka PO, Virkola NE, Pursiainen SA, Hamala IT (1988) Tappi J 71(2): 51
62. Simons HA, AF-IPK-Multi-client study (1992) Towards Kraft Mill 2000 Vancouver, Book 2,
p 54
63. Nutt WE (1992) Presented in the Ninth Sunds Defibrator Technical Seminar, Williamsburg,
VA, USA
64. Griggs BF, Eachus SW, Joseph JC (1992) Annual Meeting Technical Section, CPPA,
Montreal, Quebec, Canada, Book B, p 209.
65. Meadows DG (1993) Tappi J 75(1): 71
66. Tatsuishi H, Hatano T, Iwai T, Kovasin K (1987) Internat Oxygen Delignification Conf Proc,
Tappi Press, Atlanta, p 209
67. Brannland R, Fossum G (1987) Internat Oxygen Delignification Conf Proc, Tappi Press,
Atlanta, p 59
68. Enz S, Emmerling F (1987) Internat Oxygen Delignification Conf Proc, Tappi Press, Atlanta,
p 13
69. Jones AR (1983) Tappi J 66(12): 42
70. Liebergott N, Van Lierop B, Garner BC, Kubes GJ (1984) Tappi J 67(8): 76
71. Liebergott N, Van Lierop B (Nov. 1978) Tappi Oxygen, Ozone and Peroxide Pulping and
Bleaching Seminar, New Orleans, Louisiana
72. Liebergot N, Vanlierop B, Fleming BI (1992) Bleaching vol 2, Tappi Press, Atlanta, Georgia,
p 737
73. Sixta H, Hoglinger O, Gotzinger G (1989) In: Proc of 1989 ISWPC Symp, North Carolina
State University, Raleigh, North Carolina, May 22-25, p 387
74. Wartiovaara I, Blomberg L (1986) In: Proc of EuCePa Symp Environmental Protection in the
90's, Helsinki, Finland, May 19~2, p 110
75. Axegard P (1986) Tappi J 69(10): 54
76. Axegard P (1986) Pulp and Paper Sci 12(3): J76
77. Hakulinen R, Salkinoja-Salonen M (1982) Internat Pulp Bleaching Conf Proc, Tappi press,
Atlanta, p 97
78. Sjoblom K, Hardmeier P (1988) Internat Pulp Bleaching Conf Proc, Tappi Press, Atlanta,
p 263
79. Kortelainen VA, et al. (1982) Internat Pulp Bleaching Conf Proc, Tappi Press, Atlanta
80. Nasman L (1981) Pulp and Paper 55(10): 137

81. Lachenal D, De Chonclens C, Boursen L (1986) Tappi J 69(7): 90
82. Torregrossa LO, Kortelainen VA, Gullichsen J (1985) Pulping Conf Proc, Tappi Press,
Atlanta, p 601
83. Godsay MP, Pearce EM (1984) Oxygen Delignification Conf Proc, San Francisco, CA, p 55
84. Strunk WG (1983) Pulp Paper Can 84(4): 53
85. Carre G, Nasman L, Annergren G, Lindstrom L (1982) Internat Pulp Bleaching Conf Proc,
Tappi Press, Atlanta, p 17
86. Lachenal D, Bourson L, DeChoudens C (1985) Pulping conf Proc, Tappi Press, Atlanta, p 439
87. Hastings C, Fredstrom C, Idner K, Proc of the 1992 Non-chlorine Bleaching Conf, Hilton
Head, SC, USA
88. Vidal T, Colom JF (1992) Tappi J 75(7): 213
89. Hayos M, Robberechts M, Seccombe R, Nugent AJ, Walsh PB (1992) Proc of the 1992 Appita
Conf, Launceston, Australia, p 69
90. Bursey R, Sinclair WH, Heimburger SA (1992) Presented at the 1992 CPPA Pacific Coast
Branch Conf, Canada
91. Bugajer S, Danilas RM (1987) Pulp Paper Can 88(12): 169
92. Carre G (1992) Presented at the 9th Sunds Defibrator Technical Seminar, Williamsburg, VA,
USA
93. Basta J, Holtinger L, Lundgren P, Fasten H (1991) In: Proc of the Internat Pulp Bleaching
Conf, Stockholm, Sweden, vol 3, p 23
94. Liebergott N, et al. (1983) Ozone Sci Eng 4(3): 109
95. Liebergott N (1988) Technical Presentation at Spring Meeting of Tappi Bleaching Conf
Committee, Pensacola, FL
96. Schwarzl K, Presented at the 1991 NCSU Workshop on Emerging Pulp and Chlorine-Free
Bleaching Techniques, Raleigh, NC, USA
97. Igerud L (1992) Presented at the Ninth Sunds Defibrator Technical Seminar, Williamsburg,
VA, USA
98. De Ruvo A (1985) In: Proc of 1985 ISWPC Symp, Aug. 26 30, Vancouver, British Columbia,
Canada
99. Andersson KA (1977) Tappi J 60(3): 95
100. Lindberg S, Lund LB (1980) Tappi J 63(3): 65
101. Dorica J, Wong A, Garner BC (1986) Tappi J 69(5): 122
102. Dorica J, Wong A, Garner BC (1985) Pulping Conf Proc, Tappi Press, Atlanta, p 587
103. Reeve DW, Rapson WH (1970) Pulp and Paper Mag Can 71(13): T274
104. Rapson WH (1965) Pulp Paper Mag Can 66(4): T-295
105. Rapson WH, Reeve DW (1973) Tappi J 56(9): 13
106. Eriksson K-EL (1990) Wood Sci Technol 24:79
107. Paice MG, Bernier R, Jr, Jurasek L (1988) Biotech. Bioeng 32:235
108. Viikari L, Sundquist J, Kettunen J (1991) Paper and Timber, 384
109. Kantelinen A, Hortling B, Sundquist J, Linko M, Viikari L (1993) Holzforschung, 47:318
110. Paice MG, Gurnagul N, Page DH, Jurasek L (1992) Enzyme Microb Technol 14:272
111. Clayton DW, Stone JE (1967) Pulp and Paper Mag Can 64:T459
112. Munk N, Bleach boosting with xylanases: recent research results (1993) In: Proc of 47th Appita
Annual General Conf, Rotorua, New Zealand, vol 1, p 257
113. Bajpai P, Bhardwaj NK, Maheshwari S, Bajpai PK (1993) Appita 46(4): 274
114. Bajpai P, Bhardwaj NK, Bajpai PK, Jauhari MB (1994) J Biotech 36:1
115. Bajpai P, Bajpai PK (1996) Tappi J 79(4): 225
116. Viikari L, Ranua M, Kantelinen A, Linko M, Sundquist J (1987) In: Proc of4th Internat Symp
on Wood & Pulping Chemistry, Paris, vol 1, p 151
117. Viikari L, Ranua M, Kantelinen A, Sundquist J, Linko M (1986) Bleaching with enzymes In:
Proc of Third Internat Conf on Biotechnol in Pulp and Paper Industry, Stockholm, Sweden, p 67
118. Viikari L, Kantelinen A, Sundquist J, Linko M (1994) FEMS Mircobiology review, 13:335
119. Clark TA, Steward D, Bruce ME, McDonald AG, Singh AP, Senior DJ (1991) In: Proc of 45th
Appita Annual General Conf, Melbourne, vol 1, p 193
120. Buchert J, Siika-aho M, Kantelinen A, Ranua M, Viikari L (1992) 5th Int Conf Biotech in the
Pulp and Paper Industry, Kyoto
121. Skerker PS, Farell RL (1991) In: Proc of Internat Pulp Bleaching Conf, Stockholm, Sweden,
vol 2, p 93
122. Sinner M, Ditzelmuller G, Wizani W, Steiner W, Esterbauer E (1991) VAI-Biobleiche Papier

45:403
123. Perrolaz JJ, Davis S, Gysin B, Zimmerman W, Casimir J, Fiechter A (1991) In: Proc of 6th
Internat Symp on Wood and Pulping Chemistry, Melbourne, vol 1, p 485
124. Bajpai P, Bajpai PK (1996) Advances in Biochemical Engineering/Biotechnology, vol 56:
Chap 1, p 1
125. Viikari L, et al. (1996) Adv. Biochemical Engineering/Biotechnology, 57: ?
126. Bourbonnais R, et al. (1990) FEBS Lett 267:99
127. Olsen WL, et al. (1989) EP 345 715
128. Egan M (1985) In: Second Annual Pulp and Paper Chemical Outlook Conf November 12 13,
Montreal, Corpus Information Services Ltd., Montreal
129. Viikari L, Tenkanen M, Buchert J, R/itto M, Bailey M, Siika-aho M, Linko M (1993) In:
Bioconversion of Forest and Agricultural Plant Residues (Saddler JN ed) CAB International,
Wallingford, p 131
130. Arbeloa M, Leseleac JD, Goma G, Pommier JC (1992) Tappi J 75(3): 215
131. Arbeloa, M, Leseleac JD, Goma G., Pommier JC (1992) Eur Patent Appl 49667
132. Archibald F (1992) Holzforschung 4:305
133. Dodson PJ, Evans CS, Harvey PJ, Peliner JM (1987) FEMS Microb Lett 42:17
134. Johansson T, Nyman PO (1987) Acta Chem Scand B41:762
135. Archibald FS (1992) Appl Environ Microbiol 58:3101
136. Perie FH, Gold MH (1991) Appl Environ Microbiol 57:2240
137. Galliano H, Gas G, Boudet A (1988) Plant Physiol Biochem 26:619
138. Haemmerli SD, Leisola MSA, Feichter A (1986) FEMS Microb Lett 35:33
139. Hammel KE, Moen MA (1991) Enzyme Microb Technol 13:15
140. Ander P (1990) In: Advances in Biological Treatment of Lignocellulosic Materials (Coughlan
MP, Amaral Collaco MT, eds), p 287, Elsevier, New York
141. Call HP (1987) EP 327 576
142. Call HP (1990) DE 40 08 893
143. Call HP (1992) WO 92/20857
144. Call HP, Mucke I (1994) Proc of Nonchlorine Bleaching Conf
145. Call HP, Mucke I (1995) Internat Nonchlorine Bleaching Conf, Amelia Island, Florida, U.S.A.
146. Call HP, Mucke I (1995) 6th Internat Conf on Biotech in Pulp and Paper Industry, Vienna, Austria
147. Kondo R, Hirai H, Harazono K, Sakai K (1995) 6th Internat Conf on Biotech in Pulp and
Paper Industry, Vienna, Austria
148. Paice MG, Bourbonnais R, Reid ID (1995) 6th Internat Conf on Biotech in Pulp and Paper
Industry, Vienna, Austria
149. Ducka I, Pekakrovicova A (1995) 6th Internat Conf on Biotech in Pulp and Paper Industry,
Vienna, Austria
150. Reid ID, Paice M G (1994) FEMS Microbiology Reviews 13:369
151. Tran AV, Chambers RP (1987) Appl Microbiol Biotechnol 25:484
152. Kirkpatrick N, Reid ID, Ziomek E, Ho C, Paice M (1989) Appl Environ Mircobiol 55:1147
153. Kirkpatrick N, Reid ID, Ziomek E, Paice M G (1990) Appl Environ Microbiol 33:105
154. Fujita K, Kondo R, Kokki S (1991) Tappi J 74(11): 123
155. Faix O, Muzuch MD, Kirk TK (1985) Holzforschung 39:203
156. Leisola M, Brouwn C, Laurila M, Ulmer D, Fiechter A (1982) Eur J Appl Microbiol Biotechnol
15:180
157. Reid ID, Paice MG, Ho C, Jurasek L (1990) Tappi J 73(8): 149
158. Adler E, (1977) Wood Sci Technol 11:169
159. Ni Y, Kubes GJ, Van Heiningen ARP (1990) J Pulp and Paper Sci 16(1): J13
160. Kirk TK, Yang HH (1979) Biotech Lett 1:347
161. Pellinen J, Abuhasan J, Joyce TW, Chang HH (1989) J Biotech 10:161
162. Ander P, Eriksson KE (1976) Material U Organismen 3:129
163. Dawson-Andoh BE, Morrel JJ, Biermann CJ, Hull JL (1991) Tappi J 74(10): 187
164. Wroblewska H, Zielinsk MH (1995) 6th Internat Conf on Biotech in Pulp and Paper Industry,
Vienna, Austria
165. Pazukhina GA, Soloview VA, Malysheva ON (1995) 6th Internat Conf on Biotech in Pulp and
Paper Industry, Vienna, Austria
166. Nishida T, Katagiri N, Tsustsumi Y (1995) 6th Internat Conf on Biotech in Pulp and Paper
Industry, Vienna, Austria
167. Kirk TK (1976) Connors WJ, Zeikus JG (1976) Appl Enviorn Microbiol 32:192
168. Reid ID (1979) Can J of Botany 57:2050
169. Yang HH, Effland M J, Kirk TK (1980) Biotechnol Bioeng 22:65
170. Eriksson KE, Hamp SG (1978) Eur J Appl Microbiol Biotechnol 10:183
171. Eriksson KE, Goodell EW (1974) Can J Microbiol 20:371
172. Kirkpatrick N, Reid ID, Ziomek E, Paice M G (1990) In: Biotech in Pulp and Paper Manufac-
ture Applications and Fundamental Investiagations (Kirk TK, Chang HM eds.), Butterworth
- Hienemann, Toronto, p 131
173. Norris DM (1980) Appl Environ Microbiol 40:376
174. Milstein O (1981) Eur J Appl Microbiol Biotechnol 13:117
175. Jeffries TW, Choi S, Kirk TK (1981) Appl Environ Microbiol 42:290
176. Keyser P, Kirk TK and Zeikus JG (1978) J Bacteriol 135:790
177. Tien M, Kirk TK (1983) Science, 221:661
178. Leatham GF, Kirk TK (1983) FEMS Microb Lett 16:65
179. Hatakka AI, Buswell JA, Pirhonen TI, Uusi-Rauva AK (1990) In: Proc Biotech in the Pulp and
Paper Industry, London, p 152
180. Boyle CD, Bradely RK, Reid ID (1992) Appl Environ Microbiol 58:3217
181. Janshekar H, Fiechter A (1983) In: Adv in Biochem Eng vol 27, Pentoses and Lignin (Fiechter
A, ed), Springer, Berlin Heidelberg New York
182. Kirk TK, Yang HH, Keyser P (1978) Dev Ind Microbiol 19:51
183. Drew S, Kadam KL (1979) Develop Indust Microbiol 20:153
184. Kirk TK, Schultz E, Connors WJ, Lorenz LF, Zeikus JG (1978) Arch Microbiol 117:277
185. Fenn P, Kirk TK (1979) Arch Microbiol 123:307
186. Bar-Lev SS, Kirk TK (1981) Biochem Biophys Res Comm 99:373
187. Reid ID, Chao EE, Dawson PSS (1984) Can J Microbiol 31:88
188. Reid ID, Paice MG (1992) In: Leatham G F (ed) Frontiers in Industrial Mycology, Chapman
and Hall, New York, p 112
189. Reid ID, Seifert KA (1982) Can J of Botany 60:252
190. Ziomek E, Kirkpatrick N, Reid ID (1991) Appl Microbiol Biotechnol 35:669
191. Kirkpatrick N, Ziomek E, Reid ID (1990) In: Kirk TK, Chang HM (eds) Biotech in Pulp and
Paper Manufacture applications and Fundamental Investigations Butterworth-Heinemann,
Toronto, p 137
192. Goldsby GP, Enoki A, Gold MH (1980) Arch Microbiol 128:190
193. Weinsten DA, Krisnangkura K, Mayfield MB, Gold MH (1980) Appl Environ Microbiol 39:535
194. Jager A, Croan S, Kirk TK (1985) Appl Environ Microbiol 50:1274
195. Paice MG, Jurasek L, Ho C, Bourbonnais R, Archibald F (1988) Tappi J 72(5): 217
196. Leisola MSA, Fiechter A (1985) FEMS Microb Lett 29:33
197. Kirkpatriek N, Palmer JH (1987) Appl Microbiol Biotechnol 27:129
198. Jurasek L, Paice MG (1988) Biomass 15:103
199. Ho C, Jurasek L, Paice M G (1990) J Pulp and Paper Sci 16(5): J78
200. NASCI (1969) Technical Bull No 228
201. Gould M (1970) US Patent, No 3,531,370
202. EPA (1982) Final Development Document for Effluent Limitations Guidelines and Standards
for the Pulp, Paper and Paperboard Point Source Category, EPA 440/1 82/025
203. Schmidt RL, Joyce TW (1980) Tappi J 63(12): 63
204. Fuller RR, Williams HH, Hodge G (1976) Tappi J 59(9): 66
205. Almemark M, Ekengren O (1989) In: Proc of the 5th ISWPC Symp, NCSU, Raleigh, NC, p 739
206. Milstein O, Haars A, Majcherczyk A, Trojanowski J, Tautz D, Zanker H, Huttermann A (1987)
In: Proc of the Second IAWPRC Symp on Forest Industry Wastewaters, Tampere, Finland
207. NACSI (1980) Technical Bull No 340
208. Watson JB, Lanza GR (1984) 1984 R&D Conf Proc, Tappi Press, Atlanta, p 193
209. Lassa JM (1981) Tappi J 64(3): 181
210. Carter CN, Sigler RG (1981) Tappi J 64(9): 135
211. Rock SL, Alexander B, Kennedy DC (1974) Tappi J 57(9): 87
212. Borjeson HB, Lindberg S (1981) Tappi J 64(10): 89
213. Lundahl H, Mansson I (1980) Tappi J 63(4): 97
214. Swaney JM (1985) Pulp and Paper Can 86(2): 38
215. Wallace AT, Grimestad G, Luoma D, Olson M (1975) In: Proc of 30th Purdue Industrial
Waste Conf, Ann Arbor Sci, Ann Arbor, p 506
216. Keenan W (1981) Paper Trade J 65(1): 63

217. Bottger J, Patzold R, Krause TH, Schempp W (1985) In: Proc of the Internat Symp on Wood
and Pulping Chemistry, Hyatt Regency Vancouver, Vancouver, BC, Canada, p 171
218. Shimada K (1982) J Japan Wood Res Soc (Mokuzai Gakkaishi) 28(6): 376
219. Panchapakessan B, Chen CL, Gratzl JS (1989) In: Proc of 1989 ISWPC Symp Proc Post
Session, North Carolina State University, Raleigh, North Carolina, p 355
220. Sun YB, Guo HY, Joyce TW, Chang HM (1992) J Pulp and Paper Sci 18(2): J49
221. Donnini GP, Jankey SG: 1982 R&D Division Conf Proc, Tappi Press, Atlanta, p 201
222. McDonald J (1981) In: Proc of 1981 Spring Conf CPPA/TS. Pacific Cost and Western Branch,
Harrison Hot Springs, British Columbia, p 82
223. Swaney JM (1982) In: Proc of 1982 Spring Conf CPPA/TS, Pacific Coast and Western
Branches, Jasper, Alberta, p 96
224. Dogherty E (1984) Pulp Paper Can 85(1): 13
225. Servizi JA, Gordon RW (1973) Pulp Paper Mag Can 74(9): 295
226. Mueller JC, Walden CC (1974) Pulp Paper Mag Can 75(8): 274
227. Boman B, Ek M, Eriksson K-EL, Frostall B (1988) Nordic Pulp and Paper Res J 3:13
228. Rogers IH, Davis JC, Kruzynski GM, Mahood HW, Servizi JA, Gordon RW (1995) Tappi
J 58(7): 136
229. Leach JM, Mueller JC, Walden CC (1977) CPAR Report, No 408-2, Canada Forestry Service,
Ottawa, Ontario
230. Gergov M, Priha M, Talka E, Valltila O, Kangas A, Kukkonen K (1988) 1988 Environ Conf
Proc, Tappi Press, Atlanta, p 443
231. Voss R, Wearing JT, Wong A (1981) In: Keith LH (ed) Advances in the Identification and
Analysis of Organic Pollutants in Water, Ann Arbor Science, Ann Arobor, vol 2, p 1059
232. Deardorff TL, Willhelm RR, Nonni AJ, Renard JJ, Phillips RB (1994) Tappi J 77(8): 163
233. Jokela JK, Laine M, Ek M, Salkinoja-Salonen M (1993) Environ Sci Technol 27:547
234. Graves JW, Joyce TW, Jameel H (1993) Tappi J 76(7): 153
235. Lafond RA, Ferguson JF (1991) Tappi Proc 1991 Environ Conf, p 797
236. Ek M, Eriksson KE (1987) In: Proc of 1987 ISWPC Symp, Paris, France, p 128
237. Vargas C, Ahlert RC (1987) J Wat Pol Can Fed 59:964
238. McFarlane PN, Greenwood JM (1987) In: Proc of 1987 ISWPC Syrup, Paris, France, p 124
239. Gunnarsson L, Rosen B, Ronner U, (1987) In: Proc of 1987 ISWPC Symp, Paris, France
240. Andersson PE, Gunnarsson L, Welander T, Wikstrom A, (1987) In: Proc of the 4th ISWPC
Symp, Paris, France
241. Wilkinson SG (1967) J Gen Microbiol 47:67
242. Walden CC, Howard TE (1981) Pulp Paper Can 82(4): 115
243. Raizer-Neto E, Pichon M, Rouger J (1989) In: Proc of the 4th Internat Biotech Conf in
Pulp and Paper Industry, Abstract of Papers, Mission Valley Inn, Raleigh, North Carolina,
p71
244. Fitzsimons R, Ek M, Eriksson KE (1989) In: Proc of the 4th Internat Biotech Confin Pulp and
Paper Industry, Abstract of Papers, Mission Valley Inn, Raleigh, North Carolina, p 75
245. Macalady DL, Trantnyek PG, Grundl TJ (1986) J Contaminant Hydrology 1:1
246. Armenante PM, KafKewitz D, Lewandowski G, Kung CM (1992) Environ Progress 11:113
247. Strehler A, Welander T (1994) Wat Sci Tech, vol 29, No 5 6, 295
248. Campbell AG, Jr (1983) "A Bench Scale Evaluation of the MyCoR Process for Decolorization
of Bleach Plant Effluent Using the White-rot Fungus Phanerochaete chrysosporium", PhD
Dissertation, North Carolina State University, Raleigh, NC
249. Chang HM, Joyce TW, Matsumato Y, Yin CF, Boechat CA, Kirk TK (1986) In: Proc of the 3rd
Internat Conf on Biotech in the Pulp and Paper Industry, Stockholm, Sweden, p 120
250. Royer G, Desrochers M, Jurasek L, Bouleau D, Mayer RC (1985) J Chemical Technol
Biotechnol 35:14
251. Kirk TK, Chang HM (1975) Holzforschung 29:56
252. Crawford DL, Crawford RL (1980) Enzyme Microbiol Technol 2:11
253. Sundman G, Kirk TK, Chang HM (1981) Tappi J 64(9): 145
254. Yin CF, Joyce TW, Chang HM (1989) In: Proc of the Industrial Waste Conf, Lafayette W,
Indiana, Purdue University
255. Yin CF, Joyce TW, Chang HM (1989) J Biotechnol 10:67
256. Huynh VB, Chang HM, Joyce TW, Kirk TK (1984) In: Proc of Tappi R&D Conf Proc, Sept,
Appleton, WI, p 185
257. Guo HY, Chang HM, Joyce TW, Glasser JH (1989) In: Proc of Internat Biotech Confin Pulp
and Paper Industry, Abstract of Papers, Mission Valley Inn, Raleigh, p 69
258. Chang HM, Vasudevan B, Joyce TW, Taneda H (1987) In: Proc of Annual Meeting of the
Amer Chem Soc, New Orleans, LA
259. Eaton DC (1985) Enzyme Microb Technol 7:194
260. Bumpus J, Tien M, Wright D, Aust S (1985) Science 228:1434
261. Eaton D, Chang HM, Kirk TK (1980) Tappi J 63(10): 103
262. Kirk TK, Farrel RL (1976) Annu Rev Microbiol 33:192
263. Reddy CA, Forney LJ, Kelly RL (1983) In: Recent Advances in Lignin Biodegradation
Research, Uni Publ, Tokyo, p 153
264. Kirk TK (1978) Arch Microbiol 117:277
265. Kirk TK, Shimada M (1985) In: Higuchi T (ed) Biosynthesis and Biodegradation of Wood
Components, Academic Press, San Diego, CA, p 579
266. Eaton DC, Chang HM, Joyce TW, Jeffries TW, Kirk TK Tappi J 65(6): 89
267. Campbell AG, Gerrard ED, Joyce TW, Chang HM, Kirk TK (1982) Proc of the Tappi
Research and Development Division Conf, Asheville, NC, p 209
268. Huynh VB, Chang HM, Joyce TW, Kirk TK (1985) Tappi J 68(7): 98
269. Pellinen J, Yin CF, Joyce TW, Chang HM (1988) J Biotechnol 8:67
270. Yin CF, Joyce TW, Chang HM (1989) J Biotechnol 10:67
271. Pellinen J, Joyce TW, Chang HM (1988) Tappi J 71(9): 191
272. Yin CF, Joyce TW, Chang HM (1989) In: Proc of the 4th Internat Biotech Conf in Pulp and
273. Messner K, Ertler G, Farcher S (1989) In: Proc of the 4th Internat Biotech Conf in Pulp and
274. Livernoche D, Jurasek L, Desrochers M, Dorica J (1983) Biotechnol Bioeng 25:2055
275. Royer G, Livernoche D, Desrochers M, Jurasek L, Rouleau D, Mayer RC (1983) Biotechnol
Lett 5(5): 321
276. Royer G, Desrochers M, Jurasek L, Rouleau D and Mayer RC (1985) J Chem Technol
Biotechnol 35B, 19
277. Bergbauer M, Kraepelin G (1989) In: Proc of the 4th Internat Biotech Confin Pulp and Paper
Industry, Abstract of Papers, Mission Valley Inn, Raleigh, North Carolina, p 79
278. Bergbauer M, Eggert C, Kraepelin G (1991) Appl Microbiol Biotechnol 35:105
279. Bajpai P, Mehna A, Bajpai PK (1993) Process Biochem 46:274
280. Mehna A, Bajpai P, Bajpai PK (1995) Enzyme Microb Technol 17:18
281. Royer G, Yerushalmi L, Rouleau D, Desrochers M (1991) J Indl Microbiol 7:269
282. Archibald F, Paice MG, Jurasek L (1990) Enzyme Microbiol Technol 12:846
283. Marton J, Stern AM, Marton T (1969) Tappi J 52(10): 1975
284. Roy-Arcand L, Archibald FS (1991) Tappi J 74(9): 211
285. Belsare DK, Prasad DY (1988) Appl Microbiol Biotechnol 28:301
286. Fukuzumi T (1980) In: Kirk TK, Chang HM, Higuchi T (eds) Lignin Biodegradation vol 2,
CRC Press, Boca Raton, FL, p 161
287. Kannan K (1990) World J Microbiol Biotechnol 6, 114
288. Tono T, Tani Y, Ono KJ (1968) Ferment Technol 46:569
289. Milstein O, Haars A, Majcherczyk A, Trojanowski J, Tautz D, Zanker H, Huttermann A
(1987) In: Proc of the Second IAWPRC Symp on Forest Industry Wastewaters, Tampere,
Finland
290. Prasad DY, Joyce TW (1991) Tappi J 74(1): 165
291. Schwarts RD, Hutchinson DB (1981) Enzyme Microb Technol 3:361
292. Bollag JM, Liu SY, Minard RD (1979) Appl Environ Microbiol 38:90
293. Klibanov AM, Morris ED (1981) Enzyme Microb Technol 3:119
294. Ruggiero P, Sarkar JM, Bollag JM (1989) Soil Sci 147:361
295. Bollag JM, Shuttleworth KL, Andersson DH (1988) Appl Environ Microbiol 54:3086
296. Topp E, Crawford RL, Hanson RS (1989) Appl Environ Microbiol 54:2452
297. O' Reilly KT, Crawford RL (1989) Appl Environ Microbiol 55:2113
298. Mikesell MD, Boyd SA (1986) Appl Environ Microbiol 52:861
299. Apajalahti JHA, Salkinoja-Salonen MS (1987) Bacteriol 169:5125
300. Struijs J, Rogers JE (1989) Appl Environ Microbiol 55:2527
301. Lyr VH (1963) Phytopathol Z 47:73
302. Hammel KE, Tardone PJ (1988) Biochem 27:6563
303. Arcand LR, Archibald FS (1991) Enzyme Microb Technol 13:194

304. Paice MG, Jurasek L (1984) Biotech Bioeng 26:477
305. Forss K, Jokinen K, Savolainen M, Williamson H (1987) Fourth Internat Symp on Wood and
Pulping Chemistry, Paris, p 179
306. Call HP (1992) EP 92/01086
307. Ferrer I, Dezotti M, Duran N (1991) Biotechnol Lett 13:577
308. Farrel RL (1987) WO 87/00564
309. Farrel RL (1987) Philos Trans R Soc London A 321:549

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Reduction of Organochlorine Compounds

in Bleach Plant Effluents

Pratima Bajpai and Pramod K. Bajpai

List of Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

ADI Acceptable daily intake

(DC) Treatment by mixing chlorine dioxide and chlorine simultaneously

Table 1. Regulations for discharge of chlorinated organic compounds

1.2 ] Scandinavian mills

Fig. 1. AOX emissionsfrom Scandinavianand someCanadianbleachingkraft pulp millsin 1994

Relatively lipophilic Log Pow > 3

formation of TOC1 and chlorinated phenolics increased with increasing temper-

2 Environmental Impact of Chlorinated Organic Compounds

About 300 different chlorinated organic compounds in bleached pulp mill

Table2. Chlorinated organic compounds in bleached pulp mill effluents

Table 3. Polychlorinatedphenoliccompoundsproposed for regulation

Table 4. Bleaching processes in order of decreasing environmental effect

S.no. a Bleaching process Comments

1 (C90DIo) E1HD1E2D2 Traditional technique

aSoftwood Kraft Pulp

invertebrate density and parasitic infestation of stationary fish species. Even at

the bleaching. It is a well-known herbicide and biocide, especially for brown

3 Reducing the Generation of Organochlorine Compounds

The emission of chlorinated organic compounds from bleach plants can

3.1 Enzymatic Prebleaching

Pretreatment of pulp with enzymes prior to bleaching helps in substantially

3.1.2 Ligninolytic enzymes

fungi secrete a number of oxidative enzymes and some hitherto unknown

Table 5. Effects of lignin peroxidase on kappa number and bright-

After treatment After bleaching

Kappa Brightness Brightness

Conditions of enzyme treatment: pH 3, temperature 30 ~ veratryl

Recently, Hammel and Moen [139] reported depolymerization of a synthetic

Table 6. Differencesbetweenxylanaseand laccase/mediatortreatment

fibres. A repeated enzymatic treatment is possible and results in a 50-70%

3.2 Fungal Prebleaching

Table 7, Change in brightness of hardwood kraft pulp after exposure to various

C. versicolor was found to be ineffective in brightening softwood kraft

source such as glucose or cellulose to metabolize lignin to form carbon dioxide

pulp [191]. Together with C. versicolor they acted synergistically, resulting in an

4 Treatment of Bleach Plant Effluents

4.1 Biological Treatment

Biological treatment is known to be effective in reducing the BOD and simulta-

4.1.1 Using Bacteria

4.1.1.2 Anaerobic Processes. Anaerobic biological treatment can also efficiently

E K and Eriksson proposed a process based on U F and anaerobic and

4.1.2 Using Fungi

markedly affected by oxygen concentration [-261-265]. The fungal decoloriz-

Fig. 3. Rotatingbiologicalcontactorfor fungaldecolorizationof pulp mill effluent

ii! ---- ='" ~

Guo et al. [257] investigated degradation of model compounds - chloro-

4.2 Enzymatic Treatment

Table 9. Removalof chlorophenoliccompoundsfrom a mixture in high-molecular-mass

0 ~0 0 0 ' ~ . ~ ;> ~ ~'~

Chlorinated phenols 141 9 93.6

P. chrysosporium m u t a n t strain SC 26 a n d d e s c r i b e d in 2 p a t e n t s assigned to the

M a n y process c h a n g e s have been i m p l e m e n t e d o r are being c o n s i d e r e d to

1. Rennel J (1995) Nordic Pulp and Paper Res J 1:34

3. Lindberg S (1974) Chem Abs 80 1,407, 404m

80. Nasman L (1981) Pulp and Paper 55(10): 137

122. Sinner M, Ditzelmuller G, Wizani W, Steiner W, Esterbauer E (1991) VAI-Biobleiche Papier

216. Keenan W (1981) Paper Trade J 65(1): 63

303. Arcand LR, Archibald FS (1991) Enzyme Microb Technol 13:194

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