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Cellulosic and Tannins Containing Wastewater Treatment Using MBBR
Technology and Fungal Strain
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International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
IOP Publishing
doi:10.1088/1757-899X/877/1/012056
1
Cellulosic and Tannins Containing Wastewater Treatment
Using MBBR Technology and Fungal Strain
I C Moga1*, O G Iordache2, G Petrescu1, E C Mitran2, A G Tanasa1,
I Sandulache2, G A Pantazi1, L O Secareanu2, G Anghelache1 and C Lite2
1DFR Systems SRL, Bucuresti, Romania
2National R&D Institute for Textile and Leather, Bucharest, Romania
*E-mail: corinamoga@yahoo.com
Abstract. Since the beginning, Mobile Bed Biofilm Reactor (MBBR) technology has been
extensively used, both at the level of small on-site treatment units and at industrial scale.
Moreover, this technology represents a starting point for many researches aimed at improving
performance, such as the use of microorganisms, enrichment with anammox bacteria to
accelerate nitrogen removal and more. Within the present paper, a new generation of carriers
(consisting of a mix of high-density polyethylene + talcum + cellulose) was bio-augmented
with a WRF (White Rot Fungi) strain, namely Cerioporus squamosus, in static conditions (data
not shown in this paper). The wastewater, targeted for treatment, originated from National
R&D Institute for Textile and Leather, INCDTP Bucharest, leather subsidiary, Leather and
Footwear Research Institute, technological flux, characterized by high tannins concentration,
and cellulosic content. Wastewater treatment aimed the reduction of COD value, as a water
quality parameter, with satisfactory results, obtaining a percentage reduction rate of 48.53%.
Also, GC-MS chromatography analysis was carried out on five vegetal tannins, used in leather
treatment, highlighting main compounds for Mimosa, Chestnut, Gambier, Myrobalan and
Quebracho natural tannins.
1. Introduction
Wastewaters containing cellulose fibers and various sources of tannins can lead to significant
changes to the physicochemical properties of these sources of wastewater, rendering them more and
more difficult to treat. Within National R&D Institute for Textile and Leather, along with its
subsidiary, Leather and Footwear Research Institute, the generated wastewaters have high content of
cellulose (due to the treatment and processing of textile materials) and tannins (due to tanning of
leather-based materials).
Cellulose is a very abundant solid material as it is the main component in wood (40-45) % and
other plant-based materials (till 90%) [1]. Cellulose is a raw material that is used in many industries
like, pulp and paper, textile, tannery, wood, biotechnological industry, pharmaceutical. In 1839 this
raw material was first segregated from plants by Anselme Payen [2]. Since then, the cellulose
extraction has been developed, and now it can be extracted from different sources (plants, wood,
tunicates, algae, bacteria etc. [3]). The global cellulose market size was 191.74 billion Euro in 2018
(Figure 1) and is projected to reach 266.47 billion Euro by 2026 [1].
International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
IOP Publishing
doi:10.1088/1757-899X/877/1/012056
2
Figure 1. Global cellulose market for 2018 [1].
Cellulose has a linear structure similar with a chain made from hundreds to thousands of linked
glucose units. The polymerization degree varies for different sources such as 10000 for cellulose
chains from nature or approx. 15000 for cellulose found in cotton [4]. In Figure 2 is presented the
chemical structure of cellulose.
Figure 2. Structure of cellulose.
Vegetable tannins are widely used in the treatment of leather materials, representing an important
stage in leather treatment, and for inducement of leather characteristic properties. Quebracho extracts
are used to produce all types of leather (bags, saddles, belts, upper shoes, garment, car seats), but its
qualities are highlighted when used to produce natural, full vegetable leather, like harnesses and
vacchetta [5]. Chestnut extract results in: gross weight yield, excellent physical and chemical
properties, even re-tanning and attractive and stable colour. In re-tanning of chrome leather, up to 30%
of chestnut extract can be added to obtain smooth, compact and full leather with a good light fastness
[6]. The application of Mimosa is very common worldwide and it can be modified by mixing with
other types of tanning materials to adopt its characteristics to the specific leather type in re-tanning.
Properties are natural soluble extracts, uniform in quality, pale in colour, low in salts and sludge
formation [7]. Myrobalans are marketed in the form of whole nuts crushed nuts or solid and spray
dried extracts wich are used in tanning of hides. Powdered myrobalans extracted with cold water are
used for dyeing red [8]. Gambier is widely used as an industrial raw material in the textile industry,
cosmetic industry, pharmaceutical industry, and as an additive to food products. In traditional
societies, gambier is used as a dye for textiles and rubber and for leather tanning. Gambier contains
natural polyphenol compounds including tannins. Complex phenolic compounds such as tannins
dissolve in polar solvents such as water (especially in hot water), methanol, ethanol, and acetone [9].
Treatment of tannins containing wastewaters is a difficult process, due to the high solubility of
these chemicals in water, and to the fact that they inhibith growth of microorganisms in activated
sludge [10-13].
International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
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doi:10.1088/1757-899X/877/1/012056
3
2. Materials and Methods
2.1. GC-MS analysis
GC-MS analysis was carried out on pure solutions of each vegetal tannin, in order to assess the main
compounds, on an Agilent Technologies 6890N spectrophotometer, with 7694E Headspace module
(Agilent Technologies) and MS 5973N MS detector. The solutions were sonicated for 10 minutes,
after which they were filtered and analyzed on gas chromatography with Mass Spectroscopy detection.
The injection of the samples was done by the automatic liquid injection mode. The method and system
parameters were as follows: Capillary column: DB-35 MS, length: 30 m, internal diameter 0,25 mm;
layer thickness: 0,25 μm; injection system: splitless; injector temperature: 300°C; constant flow: 1,8
mL/min; carrier gas Helium; temperature program: 70 °C (2 min) at 310°C with 10°C/min, 310°C (5
min); injection volume: 1,0 μl; auxiliary: 300°C; MS detector: scan mode; scanning interval: 30-
500 amu.
2.2. COD analysis
COD analysis was carried out according to SR ISO 6060, which implies the boiling with reflux for a
certain duration, of the water samples mixed with mercury sulphate (III), with a known volume of
potassium dichromate, in the presence of a silver catalyst in a strongly acidic environment (sulfuric
acid), so that part of the potassium dichromate is reduced by the oxidizable materials present. The
excess potassium dichromate is titrated with iron (III) sulphate and ammonium solution. The COD
value is calculated from the reduced amount of potassium dichromate.
For the boiling stage with reflux, a C.O.D. thermoreactor was used, namely ECO6, Velp Scientifica
type, with a temperature set to 200oC.
All reagents used to determine the chemical oxygen content were of a known analytical quality:
a) Sulfuric acid (ρ=1,84 g/mL). c(H2SO4)=4 mol/L;
b) Silver sulphate (Ag2SO4);
c) Potassium dichromate, reference standard solution. c(K2Cr2O7)=0.040 mol/L;
d) Iron (II) sulphate and ammonium, titrated solution. c[(NH4)2Fe(SO4)2 * 6H20]=0.12 mol/L;
e) Feroin, indicator solution.
The chemical oxygen consumption (COD) expressed in milligrams oxygen per liter is calculated
according to the formula (ec. 1):
COD (mg/L) =
(1)
Where: c = concentration of the amount of substance of iron (II) sulphate and ammonium solution;
V0 = the volume of the sample to be analyzed, before dilution (if performed), in milliliters;
V1 = volume of iron (II) sulphate and ammonium solution, used for titration of the control sample,
in milliliters;
V2 = volume of iron (II) sulphate and ammonium solution, used for titration of the sample to be
analyzed, in milliliters;
8000 = molar mass of ½ O2, in milligrams per liter.
2.3. Cerioporus squamosus functionalized HDPE carriers
HDPE carriers (with addition of talcum and cellulose) were functionalized, in static conditions, with a
WRF representative, namely Cerioporus squamosus. First, the strain was grown in fresh culture, on
Sabouraud nutritive broth, for 7 days, at 28oC. Afterwards, the borth, containing the fungal biomass,
was poured over HDPE carriers (which were previously sterilized at 121oC, for 15 min.) enough for
the liquid media to cover the carriers with just a few mm. The inoculated carriers were then incubated
for 10 days, at 28oC. Treatment was carried out by extraction of bio-augmented carriers, and
placement inside the testing aliquots, containing the wastewater.
International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
IOP Publishing
doi:10.1088/1757-899X/877/1/012056
4
3. Results and Discussions
Within FunCell project, textile and leather industry originated wastewater sample was subjected to
COD reduction, with the help of Cerioporus squamosus functinoalized HDPE carriers. Also, GC-MS
analysis was conducted on the sample, in order to asses tha main constituents of five tannins currently
used in leather tanning: Mimosa, Quebracho, Chestnut, Gambier and Myrobalan.
Mimosa is a vegetable tannin, which is generally extracted from Acacia mollissima bark grown in
Australia, Southeast Asia and South Africa. The bark of the tree is used for the tanning process,
because it has a very high tannin content, up to 30 percent. The skin tanned with mimosa tannin has a
reddish color. Mimosa extract is also used in natural medicine to treat headaches and diarrhea. The
general structure of the main compound is presented in Figure 3.
Quebracho is a common name in Spanish for describing the species of very hard wood trees (wood
density 0.9–1.3). The etymology of the name derives from quiebrahacha or quebrar hacha. Quebracho
produces tannins that can be extracted from the heart of both red Quebracho (Schinopsis lorentzii) [17]
and white (Aspidosperma quebracho-blanco). The general structure of the main compound is
presented in Figure 4.
Chestnut extract is obtained from the heart and the sapwood (all the young layers located between
the bark and the heart of a tree trunk, through which water and mineral salts pass) of the species of
Castanea sativa and dentata. The general structure of the main compound is presented in Figure 5.
Gambier or gambir is an extract derived from the leaves of Uncaria gambir, a shrub native to
Southeast Asia. Gambier is produced in Indonesia and Malaysia where it was an important
commercial product at the end of the 19th century. It can be used as a tanning agent, food additive
brown dye [16-17] and as a herbal medicine. Also known as pale catechu [17] or white catechu, it is
often confused with other forms of catechu (which is an extract of Acacia species, especially Acacia
catechu). The general structure of the main compound is presented in Figure 6.
Figure 3. Molecular structure
of main compound found in
Mimosa tannin.
Figure 4. Molecular structure
of main compound found in
Quebracho tannin.
Figure 5. Molecular
structure of main compound
found in Chestnut tannin.
Figure 6. Molecular structure
of main compound found in
Gambier tannin.
International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
IOP Publishing
doi:10.1088/1757-899X/877/1/012056
5
Myrobalan is the dry extract of ripe fruits of Terminalia chebula (belonging to the Combretaceae
family) and related species. These fruits are harvested from January to April and are dried by settling
in thin layers, most of them in hue.
Characteristics:
- Color: yellow-brown to brown;
- Smell: very weak;
- Taste: astringent and slightly pungent;
- Form: mucilaginous.The general structure of the main compound is presented in Figure 7.
Figure 7. Molecular structure of main compound found in
Myrobalan tannin.
Myrobalan contains about 30% of the hydrolyzable tannins, which consist of chebulinic acid,
chebulagic acid and D-galloyl glucose. Contains free tannic acid, gallic acid, ellagic acid and
mirobalanine from the resin. Due to its antiseptic and healing properties, it is used in India as a
medicine for different types of conditions such as: chronic ulcers or different types of wounds.
Commercially, it is used in the dyeing and tanning industry.
GC-MS analysis results are shown in Tables 1-5, which highlight the presence of the five tannins
used, together with their main compounds. This analysis will be the basis of future experiments on
tannin degradation in aqueous solutions.
Table 1. Chestnut tannin main compounds.
No.
Rt
CAS
Compound name
Area
1.
3.283
98-01-1
Furfural
51476
2.
5.258
97-69-8
N-Acetyl-L-alanine
5344
3.
6.046
6843-
45-4
3-
Methylimidazolidine-
2,4-dione
22109
4.
9.180
52485-
92-4
Methyl α-D-
ribofuranoside
23898
5.
9.774
4007-
18-5
3,4-dimethyl-
sydnone
12370
6.
9.871
3201-
20-5
2,4-dihydro-4,4,5-
trimethyl-3H-
pyrazol-3-one
4207
7.
10.027
125425-
35-6
Triethylene glycol
monomethyl ether
8867
8.
10.427
87-66-1
Pyrogallol
23157
9.
11.420
2595-
97-3
D-Allose
47297
10.
13.837
149-91-
7
Gallic Acid
16914
International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
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doi:10.1088/1757-899X/877/1/012056
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Table 2. Gambier tannin main compounds.
No.
Rt
CAS
Compound name
Area
1
8.571
120-80-9
Catechol
52202
2
10.238
108-46-3
Resorcinol
137335
3
11.419
87-66-1
Pyrogallol
29530
4
14.932
18979-60-7
4-Propylresorcinol
64141
Table 3. Quebracho tannin main compounds.
No.
Rt
CAS
Compound name
Area
1
8.565
120-80-9
Catechol
189877
2
10.243
108-46-3
Resorcinol
704221
3
11.414
87-66-1
Pyrogallol
41921
4
14.932
18979-60-7
4-Propylresorcinol
302142
Table 4. Mimosa tannin main compounds.
No.
Rt
CAS
Compound name
Area
1.
10.243
108-46-3
Resorcinol
639603
2.
11.419
87-66-1
Pyrogallol
109404
3.
13.070
57-50-1
Sucrose
44375
4.
13.842
498-07-7
Levoglucosan
20389
5.
14.932
451-13-8
Homogentisic Acid
63595
6.
15.655
146-72-5
3-O-methyl-D-glucose
939664
Table 5. Myrobalan tannin main compounds.
No.
Rt
CAS
Compound name
Area
1.
3.277
98-01-1
Furfural
25302
2.
10.017
67-47-0
5-hydroxymethyl furfural
90084
3.
11.414
87-66-1
Pyrogallol
251925
4.
13.076
57-50-1
Sucrose
26676
5.
13.837
498-07-7
Levoglucosan
31507
6.
15.040
33818-21-2
1,6-Anhydro-α-D-galactofuranose
10391
7.
15.579
99-16-1
Allantoic acid
11708
8.
18.666
149-91-7
Gallic Acid
564637
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IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
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doi:10.1088/1757-899X/877/1/012056
7
COD analysis is a general indicator of the water quality, measuring the capacity of dissolved
oxygen depletion, in the samples contaminated with organic matter. Specifically, the analysis
determines the equivalent amount of oxygen required for chemical oxidation of organic compounds in
water. Often, COD analysis is used to estimate BOD (Biological Oxygen Demand) values, between
these 2 indicators existing strong correlations.
The most common method of determining chemical oxygen demand is the method that involves
digestion of the sample for two hours at high temperature, under acidic conditions, in which potassium
dichromate serves as an oxidant for the organic matter present in the water samples. Silver sulphate is
present in the environment, as a catalyst, and mercuric sulfate is used to complex chlorinated
compounds, possibly present in samples. Following digestion, the degree of oxidation is determined by
indirectly measuring the oxygen requirement through the electrons consumed in reducing Cr6+ to Cr3+, by
titration techniques or spectrophotometry. The level at which the determined experimental results are
close as values to the theoretical values, first of all depends on the water load subjected to the analyzes
and on the degree of oxidation, the COD analysis strongly depending on the composition of the analyzed
water. According to the standard, a large number of organic compounds are oxidized in the proportion of
90% -100%, and if the samples contain a large amount of such compounds (the case of municipal
effluents), the COD value is a good approximation of the theoretical consumption of oxygen.
Figure 8. Acidic water
sample digestion in ECO6
thermoreactor.
The result of the analysis revealed a COD value, for the tested wastewater, of 55,849 mg/L COD
(Figure 8). Following biological treatment, with Cerioporus squamosus bio-augmented HDPE carriers
(data not shown here), it was observed a percentage reduction of 48.53% of COD content.
4. Conclusion and perspectives
MBBR technology is not currently emerging as a novel treatment technology, for conventional
wastewater, as the validates efficiency of this technology is already settled throughout the years.
However, new demands, regarding treatment of industrial wastewater, also dictates necessity of
novelty within the MBBR systems. FunCell project took a stab at obtaining novel generations of
carriers, combining HDPE with talcum and cellulose.
Furthermore, bio-agumentation of these structures was possible with fungi strains, which indeed
represents a novelty in this thematic area. Within this research paper, the authors carried out
identification of main compounds from five main vegetal tannins widely used in the leather industry.
Also, COD reduction was possible, with almost 50% reduction efficiency, from a source of natural
wastewater sample, with bio-augmented HDPE carriers, thus making a step forward validation of this
novel technology in the field of industrial wastewater treatment.
International Conference on Innovative Research - ICIR EUROINVENT 2020
IOP Conf. Series: Materials Science and Engineering 877 (2020) 012056
IOP Publishing
doi:10.1088/1757-899X/877/1/012056
8
Acknowledgements
This work was supported by a grant of the Romanian National Authority for Scientific Research and
Innovation, CCCDI – UEFISCDI, project number COFUND-MANUNET III-FUNCELL, within
PNCDI III.
5. References
[1] *** 2020, Cellulose market size, share & industry analysis, by derivate type (commodity
cellulose pulp, cellulose fibers, cellulose ethers, cellulose esters, microcrystalline cellulose,
nanocellulose, and others), by end-use industry (textile, food, chemical synthesis,
pharmaceuticals, construction, pulp & paper, paint & coating and others) and regional
forecast 2019-2026, Fortune Business Insights Publishing
[2] Castro R I and Morales-Quintana L 2019 Cellulose 26(5) 3009-3020
[3] Dong Y, Bi J, Zhu D, Meng D, Ming S, Guo W, Chen Z, Liu Q, Guo L and Li T 2019
Cellulose 26(12) 7355-7370
[4] Jiang Y, Zhang Y, Ding L, Joshua A., Wang B, Feng X, Chen Z, Mao Z and Sui X
2019 Carbohydrate polymers 223 115079
[5] Gurreiro O, Soldado D, Fialho L, Cachuco L, Garrido A, Francisco A and Jeronimo E 2019
Inclusion of rockrose and quebracho condensed tannins extracts in dairy goat diets–
influence on the production, chemical composition and fatty acid profile of milk
[6] Quave C L, Lyles J and Horswill A R 2019 U.S. Patent Application no. 16/225,281
[7] El-Sayed A, El-Sakhaw M, Kamel S, El-Gendy A and Abouzeid R 2019 Egyptian Journal of
Chemistry 62(5) 777-787
[8] Khan M A, Khan M, Srivastava P K and Mohammad F 2005 Colourage 52(12) 53-60
[9] Sy S and Kasman M 2019 IOP Conference Series: Materials Science and Engineering 546(2)
022032
[10] He Q, Yao K, Sun D and Shi B 2007 Biodegradation 18(4) 465-72
[11] Botezatu C, Duceac L D, Mastalier B, Stafie L, Jitareanu C M, Luca A C, Tarca E, Mitrea G,
Iordache A C and Patrascu T 2018 International Journal Of Medical Dentistry 22(4) 346-
350
[12] Tarnita D, Tarnita D.N, Popa D, Grecu D and Niculescu D 2010 Romanian Journal of
Morphology and embryology 51(1) 145-150
[13] Berceanu C, Tarnita D and Filip D 2010 Journal of the Solid State Phenomena, Robotics and
Automation Systems 166-167 45-50
[14] Hobbs C E 2019 Polymers 11(2) 224
[15] Gironi F and Piemonte V 2011 Chem. Eng. Res. Des. 89(7) 857–862
[16] Sakti A S, Saputri F C and Mun’im A 2019 Pharmacognosy Journal 11(1) 119-123
[17] Manalu D T and Armyanti T 2019 Agribusiness and Agricultural Economics Journal 2(1) 46-67
