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Bioresource Technology 63 (1998) 221-229

1998 Elsevier Science Ltd. All rights reserved


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0960-8524/98 $19.00
ELSEVIER PII:S0960-8524(97)00140-5

PROTEIN R E C O V E R Y D U R I N G THE OVERALL TREATMENT


OF WASTEWATERS FROM FISH-MEAL FACTORIES

L. Guerrerofl F. Omil, b* R. M 6 n d e z b & J. M. L e m a ~'


"Department of Chemical Processes, FedericoSanta Mafia Technical University, Casilla l 1O-V,Valparaiso, Chile
bDepartment of Chemical Engineering, Universityof Santiago de Compostela, Avda. das Ciencias s/n, 15706 Santiago de
Compostela, Spain

(Received 24 May 1997; revised version received 12 September 1997; accepted 17 September 1997)

Abstract discharge of these effluents can cause important


The suspended solids contained in the wastewaters environmental problems, as found in some areas like
generated by fish-meal industries (5-40g VSS/I) are Galicia (northwest of Spain) and region VIII of
composed mostly of protein, which could be recovered Chile (Omil et al., 1994; Roeckel et al., 1996).
to optimise the manufacturing process. In this work, The first process, using fish wastes, is the
settling, centrifugation and coagulation-flocculation commonest used in the European industries, such as
were studied as alternative treatment processes to those in Galicia, and such processes add value to the
attain this objective. Settling experiments showed that wastes, which are mainly constituted of residues
the flotation of a high fraction of suspended solids from tuna and sardines. The first step is the
occurred (up to 45%), even with previous alkalinisa- compression and crushing of the raw material, which
tion or the addition of a coagulant such as chitosan. is then cooked with steam, and the liquid effluent is
Centrifugation at lO000rpm for 12"5min gave filtered off in a filter press. The solids obtained are
optimum results, TSS and soluble COD removal introduced to a rotating drier and finally cut and
efficiencies of 45-82% and 10%, respectively, being crumbled to obtain the commercial fish-meal
obtained. The amino acid profile of the solids was product. The liquid stream contains oils and
quite similar to fish meal, indicating the feasibility of suspended solids (fats, proteins). This stream is
solids reuse. Results obtained after coagulation-floccu- passed through two centrifugation steps in order to
lation followed by centrifugation showed that among recover a high fraction of the suspended solids,
the different compounds assayed sodium polyacrylate which are fed back into the drier, and an effluent
and chitosan produced the best results, operating at pH rich in oils, mainly constituted by polyunsaturated
4 and 7"2-7"8, respectively, and giving TSS removal fatty acids (PUFA) of high commercial value. In
efficiencies up to 97 :and 75%, respectively. 1998 addition, an aqueous stream is generated from the
Elsevier Science Lid. All rights reserved. last centrifugation step with a high content of
organic suspended solids. This effluent, together
Key words: Centrifugation, coagulation-flocculation, with the periodical washing purges, constitutes the
fish-meal processing wastewaters, protein, reuse, main source of the wastewaters generated by these
suspended solids. factories.
The production of the wastewaters is around
5.4 m 3 per tonne of fish processed, with a variable
INTRODUCTION content in organic matter and suspended solids,
ranging from 30 to 120gCOD/1 and from 5 to
Depending on the raw material used, there are 40 g VSS/1. The final temperature is between 65 and
basically two types of fish-meal manufacturing 70C. The composition of this stream depends on
processes: those that use fish wastes, such as heads, the nature of the raw material processed in the
bones or other residues, and those that use the factory (Table 1), being mainly constituted by
whole fish. Wastewaters generated by these factories proteins, around 72%, with fats and carbohydrates
are characterised by their high organic content content around 21 and 7%, respectively. Another
(30-120g COD/I), an important part of it being specific characteristic of these wastewaters is their
suspended solids (5-40 g VSS/1). The direct high salinity, around 7-8 g C1 /1.
The second group of processes are the ones that
*Author to whom correspondence should be addressed. use the whole fish. These are widely developed in
221
222 L. Guerrero et al.

Table 1. Characteristics of the wastewaters (i) generated by the factory based on the raw material processed and (ii) used
in this work (concentrations in g/I)

Tuna bones Tuna heads Sardine Type I Type II Type III Type IV
heads
TSS 13.0 29.9 6-6 21.7 21-8 25.7 13.0
VSS 12.8 29.5 6-5 20.0 21.7 24.3 12.8
CODt 46"7 81"3 38"2 107"2 88"9 76"2 53"7
CODs 22"8 24"3 23"4 77"2 56"8 39"8 38"4
pH 6"4 6"3 6"5 6"8 6"2 6"0 6"7

Chile, one of the main fish-meal producers in the tion, it was not possible to operate always with the
world. In Chile more than 90% of the volume of fish same effluents and different wastewaters were used
captured is used for fish-meal production. The for the settling, centrifugation and coagulation-
mackerel is the most common fish used, but ancho- flocculation assays as shown in Table 1 and
vies and hake are also utilised. These processes described in the following sections.
involve the hydraulic transport of fish from ships to
factories, and the discharge, cooking and evapora- Experimental operation
tion operations. Three polluting streams arise from Three alternative operations were studied to
these processes, corresponding to the hydraulic maximise the suspended solids recovery: (a) settling,
transport of fishes (86% of total COD), the effluents (b) centrifugation and (c) the addition to coagulants/
generated during the fish reduction process by the flocculants followed by centrifugation (Fig. 1).
evaporation and deodorisation steps (8"5%) and the Results obtained from these three sets of experi-
ones resulting from the washing of the process ments were compared in terms of suspended solids
equipment and the floors of the plant (5.5%). A and soluble COD removed from the raw
complete characterisation of these wastewaters has wastewaters.
been previously reported (Roeckel et al., 1996).
The high protein content of the suspended solids
in all these wastewaters adds interest to the imple- Settling assays
mentation of a process to recover the solids in order These experiments were carried out in a 'jar-test'
to optimise fish-meal production (Marti et al., 1994) conventional set-up, with four 400ml beakers
or to use them directly as an additive for animal provided with stirring (300 rpm maximum) and time
feedstuffs. In addition, the separation of these solids control. The operational procedure was based on
can be of interest if high-rate anaerobic reactors are preliminary results and comprised the following
selected to treat these wastewaters, as anaerobic steps: (1) stirring at 100 rpm for 10 min and (2)
settling for 48 h. The characteristics of the waste-
filter or UASB reactors (Young, 1991; Lettinga and
waters used for these experiments are shown in
Hulshoff Pol, 1991). Separation would avoid opera-
Table 1 (Type I). These experiments were carried
tional problems (clogging, bulking, etc.) and
out with: (a) the raw influent, (b) after alkalinisation
minimise the possible inhibitory effects on the
of pH 8 and (c) after alkalinisation to pH 8 and
methanogenic biomass of ammonia (Omil et al.,
addition of chitosan (Fig. 1).
1995).
The aim of the present work was to study the
feasibility of removing by physico-chemical Centrifugation
processes, such as settling, centrifugation and coagu- A Beckman centrifuge (model J2-21) with tempera-
lation-flocculation, the suspended solids present in ture, speed and time controls was used. The influ-
the fish-meal processing wastewaters. The character- ences of speed (5000-15 000 rpm) and centrifugation
istics of the recovered solids were determined in time (5-15 min) were studied in order to optimise
order to study their potential reuse in fish-meal solids removal, after which the clarified effluents and
production. solids were characterised. Temperature was
maintained at 4C. The experiments were carried
out at 3000, 5000, 10000, 12000 and 15000rpm,
METHODS which corresponded to relative centrifugation veloci-
ties of 988, 2744, 10976, 15805 and 24696 g, respec-
Wastewaters tively. Once the optimum speed had been
The wastewaters were collected from a factory determined, the performance of the centrifugation
located in Galicia (Spain). Their complete character- was determined after 5, 7.5, 10, 12.5 and 15 min.
isation has been previously reported (Veiga et aL, Three types of wastewaters were used for these
1994). Due to the high variability of their composi- experiments and their characteristics are shown in
Protein recoveryfrom fish-meal wastewaters 223

/
Raw w a s~:t e w~ ~:~ '~~s

Settling experiments pH change


pH change
Addition of:
Raw Addition + NaOH - Coagulants
effluent NaOH (pH 8) - Flocculants
(pH 6.8) (pH 8) + Chitosan Centrifugation
Centrifugation

,-. . . . . . . . . . . . . . . _~. . . . . . . . . . . . . . . . . . . . . . . . . . ~.................. Y_. . . . . . . .


i
i

,, Comparison of results
I (TSS, VSS and soluble COD removal efficiencies)
i
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Fig. 1. Experimental design.

T a b l e 1 (Types II, I I I and IV). T y p e s II and I I I were Addition of coagulants/flocculants


collected f r o m the e f f l u e n t of the factory w h e n Different c o m p o u n d s w e r e used as coagulants:
washing was being u n d e r t a k e n , thus explaining their ferrous and a l u m i n i u m salts (FeCI3, FeSO4,
higher content of s u s p e n d e d solids and C O D in Fe2(504)3, A12(SO4)3), as well as an organic cationic
c o m p a r i s o n to T y p e IV. polymer, chitosan. F o r flocculation, the following

Table 2. Experimental design of coagnlation/flocculation assays (concentrations in mg/l)

Experiment Wastewater Coagulant Concentration Flocculant Concentration

AI Type III A12(804)3 100 S-2515 10


Fel Type III FeSO4 200 BL-507 10
Fe2 Type III Fe2(SO4)3 200 BL-514 10
Fe3 Type III FeCI3 200 BL-507 10
C1 Type III Chitosan 100 None --
C2 Type III Chitosan 100 S-611 10
C3 Type III Chitosan 100 S-2515 10
C4 Type III Chitosan 100 BL-514 10
C5 Type III Chitosan 100 BL-507 10
PI Type IV None -- PA-F 10
P2 Type IV None -- PA-CH 10

Table 3. Results obtained with the settling experiments (concentrations in g/I, volumes in percentage of original sample)

Raw wastewater Wastewater + NaOH Wastewater + NaOH + chitosan

Initial After settling Initial After settling Initial After settling

Top Liquid Bottom Top Liquid Bottom Top Liquid Bottom

pH 6"8 6"8 6"8 6"8 8"0 8"0 8"0 8"0 8"0 8"0 8"0 8"0
TSS 21 '7 39"6 13"7 40"9 18"8 30-6 8'7 41"7 18"7 32"9 8"3 42"8
VSS 20'0 36"6 12"2 37"4 17"5 28"5 7"6 38-5 16"8 29"6 7-5 38"5
CODt 107'2 133"3 95"3 136"3 107"2 128"3 91'5 142"1 107"7 131"8 92"8 141"8
CODs 77'2 78"4 77"1 80"2 81.1 85-5 80"1 84.5 82"5 87"3 81.6 84'1
Volume 100 24 70 6 100 15 64 21 100 14 65 21
224 L. Guerrero et al.

100%
[ ] T S S O V S S CODs
.....................
80% ..........

60% - -- -
if!i! i!i!i~ ~
:i:!:~ !ilili
40% iiii! i:::i ::::~

>
O
E 20% -- :::::I -
:::::l
r
o% iiii!il I= iiiiiiI

-20%
6.2 6.5 6.8 7.0 7.2 7.5 7,8 8.0 8.2 8.5 8.7 9.0

pH
Fig. 2. Removal efficiencies obtained after pH modification followed by centrifugation (pH of raw wastewaters was 6.2).

substances were used: S-611, S-2515, BL-514 and the optimum conditions previously determined and
BL-507 (Buckman Laboratories, Gent, Belgium). the solids and supernatants obtained were then
The combination of coagulants and flocculants used characterised. The wastewaters employed were the
in the different experiments, as well as their concen- same as those described in the previous section
trations, are shown in Table 2. In addition, sodium (Types II, III and IV).
polyacrylate (Nippon Kayaku Co. Ltd.) was used
without previous addition of coagulants. Sodium Analytical techniques
polyacrylate was used in two different forms: fine Total suspended solids (TSS), volatile suspended
particles, PA-F (Panakayaku-F sodium polyacrylate), solids (VSS), chloride and phosphate were analysed
and large particles, PA-CH (Panakayaku-CH sodium according to Standard Methods (APHA-
polyacrylate). AWA-WPLF, 1985). A semi-micro method (Soto et
The experimental procedure comprised pH al., 1989) was used to determine accurately the total
correction, addition of coagulant and stirring at and soluble COD (CODt and CODs, respectively) in
10 rpm for 10 min. When flocculants were used, the samples with high chloride concentrations. Soluble
samples were stirred after their addition at 50 rpm protein was analysed by Lowry's method (Lowry et
for 12min. After coagulant and/or flocculant al., 1951) and soluble carbohydrates were analysed
addition and stirring, samples were centrifuged at by means of the hydrazine sulphate/sulphuric

Lysine r--'m i
Fish meal
Phenylalanine

Leucine Solids recovered

Isoleucine r~ :o

Cysteine D :o

Methionine o:

Valine

Tyrosine

Alanine

Threonine

Arginine

Histidine i
Glycine

Serine
Glutamic acid

Aspartic acid

0 4 6 10 12 14 16 18 20

Composition (%)

Fig. 3. Amino acid composition (%) present in normal fish meal and in the solids obtained after centrifugation.
Protein recoveryfrom fish-meal wastewaters 225

100%

F 1~ T S S ( 3 V S S C O D s

80/ L "- I'- r-- i

il-I
'~

o%i11:
i i_l_If60%

20%
- -

--

i iii~
I :i
0% ~ - , ~ ~ ,- . . . .
Fel Fe2 Fe3 AI C1 C2 C3 C4 C5 P1 P2

Coagulant / Flocculant

Fig. 4. Removal efficiencies obtained after the addition of coagulants/ttocculants followed by centrifugation at the pH of
the raw wastewaters (detailed description of each experiment in Table 2).

method (Miller, 1959). Ammonium and redox and 12-5 min, respectively. The operation at higher
potential were determined by selective electrodes. velocities and/or longer periods of operation did not
Amino acids were determined with a hydrolysis improve the removal efficiencies attained. The
apparatus (PICO-TAG) and HPLC (Waters, Model reduction of TSS and VSS ranged between 45 and
510). 49% for the Type II wastewaters, and around
74-75% and 82-87% for Types III and IV, respec-
tively. However, the reduction of soluble COD was
RESULTS AND DISCUSSION much lower, from 7 to 11% for all wastewaters.

Settling assays
The results obtained during the settling experiments Influence of pH
showed that the formation of three different zones In order to determine the influence of pH, NaOH
occurred: a floating solids layer, the clarified liquid was added to the Type II wastewaters before centri-
and a bottom layer of solids. The characterisation of fugation. As it can be observed in Fig. 2, the
these zones is shown in Table 3. It can be seen that maximum TSS and VSS removal efficiencies were
the clarified liquid ranged from 64 to 70% of the obtained at pH 8. It can be also observed that the
total volume of the original samples. A TSS reduc- pH change promoted a slight soluble COD removal
tion of 39% of the raw wastewater was obtained and (5-18%) except at pH 8, when an increase of
it was further increased to 62% when a previous around 11% in soluble COD was observed, possibly
alkalinisation was carried out. The addition of as a result of a greater partial hydrolysis of solids
chitosan produced little improvement in these under these conditions.
results. In all experiments the formation of a super-
ficial layer with a high content of suspended solids Amino acid profile
(mainly composed of fats) occurred, although the As an important characteristic in determining the
initial alkalinisation caused a decrease of its volume potential reuse of the recovered solids in the fish-
from 24 to 15%. meal manufacturing process, the amino acid profile
These results show that, even after 48h of of the solids was compared to the values of fish
settling, it is not possible to accomplish a successful meal. As can be seen in Fig. 3, the characteristics of
separation of the suspended solids due to the flota- the recovered solids are compatible with the require-
tion of a large fraction of the solids (45% of TSS at ments for fish meal. Although there appear to be
raw pH and 24% when alkalinisation was carried some significant differences in the concentrations of
out). particular amino acids, such as glycine or phenyl-
alanine, it would not be expected that the amount of
Centrifugation solids added would significantly alter the composi-
Initial experiments showed that optimum centrifuga- tion of the fish-meal product. Based on the average
tion velocity and operational time were 10000 rpm values of the wastewaters flow and VSS concentra-
226 L. Guerreroet al.

tions, and supposing a complete suspended solids the removal efficiencies obtained with each
recovery, solids would account for only up to 8% of combination of coagulants/flocculants used. The
the total amount of fish meal produced. highest TSS and VSS removal efficiencies were
achieved when sodium polyacrylate and chitosan
Addition of coagulants/flocculants (87-92%) were used, whereas when metallic salts
Different compounds were used as coagulating and were dosed maximum removal efficiencies ranged
flocculating reagents to maximise suspended solids only from 75 to 82%. However, metallic salts
removal. The optimum pH was determined for each allowed a higher soluble COD removal efficiency
substance. (32% versus 26% with chitosan), whereas sodium
polyacrylate again produced the best results (up to
52%).
Use of coagulants/flocculants without pH modification
These experiments were carried out at the usual pH
of the raw wastewaters (6.2 for the experiments with Evaluation of optimum pH
metallic salts and chitosan, and 6.7 for the ones Based on preliminary experiments, the selected pH
carried out with sodium polyacrylate). Fig. 4 shows ranges were 4-7 for metallic salts, 4-8 for chitosan

1 0 0 % ~. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

t A

It)
80% -~ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ~ TSS .......
.2 QVSS
f-
.2 60% CODs .......
F ...............
m

tg 40%
0
E
I
Ig 20% ~ ....
i
m

0% .~ I i r --I I
3.0 4.0 5.0 6.2 6.5 6.8 7.0 7.2 7.5 7.8 . 8.0

pH

100% ~ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

B
TSS
80% __ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
I OVSS
t,- F
r

60% t . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . CODs . . . . . .
E: l
1> 40,, . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . .

0
I=:

0%
0o,o
,
oI
I ~ ::il
I, ii ...... ,..... i :!|, i:

, , ,

3.0 4.0 5.0 6.2 6.5 6.8 7.0 7.2 7.5 7.8 8.0

pH
Fig. 5. Removal efficiencies obtained after the addition of metallic salts followed by centrifugation at different pH: (a)
A12(SO4)3+S-2515; (b) FeCl3.
Protein recovery from fish-meal wastewaters 227

100%

l~ TSS [] VSS CODs


80%

, M

o 60% . . . . . . . . . . . . . . . . . !
._~
--'
400/0 - . . i ~i!ii

'
E 20% . . . . .
i!~ii
0% ', ', ', r

I
1

-20%
3.0 4.0 5.0 6.2 6.5 6.8 7.0 7.2 7.5 7.8 8.0
pH
Fig. 6. Removal efficiencies obtained after the addition of chitosan followed by centrifugation at different pH.

and 3-6.7 for sodium polyacrylate. Figures 5-7 process did not give good results, even when pH was
summarise the results obtained with metallic salts, previously modified or chitosan was added, due to
chitosan and sodium polyacrylate, respectively. the flotation of a fraction of the suspended solids
The effect of pH appears to be significant in all (up to 45%). TSS removal efficiencies obtained by
cases, the optimal range being different for each direct centrifugation for 12-5 min at 10000 rpm were
substance: 5-6 for metallic salts, 7-8 for chitosan 45-85% and these solids could be incorporated in
and 3-4 for sodium polyacrylate. This fact has to be the manufacturing process to optimise fish-meal
considered for an overall economic balance since the production because of their similar composition to
pH of the raw wastewaters usually ranges from 6 to the normal product. Treatment processes based on
7. coagulation-flocculation followed by centrifugation
The highest removal efficiencies were obtained were adequate for TSS, VSS and soluble COD
with sodium polyacrylate, either in its PA-F or removal. Among the different substances employed
PA-CH form. With this compound the reductions of (metallic salts, chitosan, sodium polyacrylate and
suspended solids and soluble COD were around 97 other flocculants), the best results were achieved
and 25-35%, respectively (Fig. 7a and b). Chitosan with sodium polyacrylate (up to 97% TSS at pH 4)
also provided high efficiencies (74-80% of TSS and and chitosan (up to 74% TSS at pH 7.2).
VSS) but in this case the highest reduction of Although sodium polyacrylate gave the best
suspended solids was obtained at pH 8 when an results both in suspended solids recovery and in the
increase of soluble COD was observed. It was not removal of soluble COD from the wastewaters, it
possible to determine if there was a solids reduction was necessary to operate at low pH, which requires
because of the coagulation process or if it was the implementation of two steps of pre-acidification
merely due to a hydrolysis (Fig. 6). However, at pH and subsequent neutralisation. This synthetic
7.2-7.8 the TSS and VSS removal efficiencies were polymer has been reported to be widely used in
remarkable (around 75%) but with only a slight some countries, such as Japan, although there is so
soluble COD removal, around 2-9%. In contrast, far no report focused on its digestibility, which is
the results obtained with metallic salts were lower essential when considering the direct use of the
than those obtained without pH modification and sludges obtained for feeding purposes. In Europe its
very different from those obtained with chitosan or use is also very common, especially for the purposes
sodium polyacrylate, ranging from 45 to 50% for that have been described in this paper, and it is
TSS and VSS. However, these substances are expected that the use of this compound will be
indicated for soluble COD removal, which ranged officially regulated in the near future.
from 13 to 20% (Fig. 5a and b). Chitosan also gave good results, with the
This work showed the feasibility of using both additional advantage that the optimum operation
centrifugation and coagulation-flocculation as was achieved at neutral pH, i.e. 7-8. Chitosan is a
operations to reduce significantly the content of derivative of chitin, the biopolymer that forms the
organic solids present in the wastewaters from fish- exoskeleton of insects and crustaceans, and is the
meal factories. The use of settling as an alternative second most abundant natural polymer after cellu-
228 L. Guerrero et al.

100%
A
I/) 80%
: r-,
................. t~TSS .....

t-. VSS
I1)
60% I~, C O D s --

40%
O
E i
i ,iii~i
20% ~ lii~ii
iili!i
:iii~
I Ii!iii:
0%
3.0 4.0 5.0 6.2 6.5 6.8 7.0 7.2 7.5 7,8 8.0
pH

100%
B
............. ~TSS .......
I/) 80%
o DVSS
P
I1}
"5 60% ............... CODs . . . . . .

N> 40%
0
E
20%

0% ..... i .....t t I I i

3,0 4.0 5.0 6.2 6.5 6.8 7.0 7.2 7,5 7,8 8.0
pH
Fig. 7. Removal efficiencies obtained after the addition of sodium polyacrylate followed by centrifugation at different pH:
(a) PA-F; (b) PA-CH.

lose. Its use as cattle feedstuff has been suggested, in which the soluble organic matter would be effect-
among other potential applications (Rawis, 1984). ively removed.
Buelna et al. (1990) reported the use of chitosan for
the recovery of algal biomass grown on pig excre-
ment for its use as animal feedstuff, both alone or in ACKNOWLEDGEMENT
combination with synthetic polymers. They reported
removal efficiencies of 85% with 10 mg/l of chitosan This work was supported by the Spanish Commis-
at pH 7, which could be increased with the use of sion of Science and Technology (CICYT), Project
synthetic polymers. The potential use of these AMB95-0365.
sludges for feeding purposes clearly indicates the
'clean' nature of these process.
Apart from the feasibility of both centrifugation REFERENCES
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reduction of suspended solids, this treatment makes APHA-AWA-WPCF (1985). Standard Methods for the
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reactors, such as anaerobic filters or UASB reactors, E. P. (1990). Evaluation of various flocculants for the
Protein recoveryfrom fish-meal wastewaters 229

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Wastes, 31, 211-222. chitin wastes to valuable products. Chem. Eng. News, 62,
Lettinga, G. & Hulshoff Pol, L. W. (1991). UASB-process 42-45.
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Technol., 24, 87-107. clean technology in the fish-meal industry by addition of
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J. (1951). Protein measurements with the folin phenol 96-104.
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Marti, C., Roeckel, M., Aspe, E. & Kanda, H. (1994). (1989). Semimicro COD determination method for high
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