RESULTS AND DISCUSSION
4.1 PERFORMANCE OF SEQUENTIAL ANAEROBIC–ANOXIC–AEROBIC CONTINUOUS MOVING BED REACTOR (CMBR) SYSTEM
4.1.1.2 Performance of anoxic CMBR (R2) at varied influent thiocyanate
Table 4.1 (b): Average performance of anaerobic reactor (R1) at feed SCN– concentration variation
SCN– pH VFA
Feed (mg/L)
Biogas (mL/day)
SMA (g CH4–COD/
g VSS. day)
Se
TVS (mg/L)
Se
0 344 0.349 6.7 12600 379 (14)
110 156 0.102 6.8 10200 175 (4)
200 56 0.036 6.9 11000 288 (45)
450 0 ND 6.8 10750 194 (40)
600 0 ND 6.7 10500 285 (31)
Se: Effluent (mg/L), ND: not detectable; VFA: Volatile fatty acid as acetic acid (mg/L) Numbers in parenthesis indicate standard deviation values
Total volatile solids (TVS) in R1 was almost 12600 mg/L in absence of thiocyanate and decreased little to 10,000–11,000 mg/L with addition of SCN– then remained stable at 10700– 10500 mg/L at maximum feed SCN–. The ratio of attached biomass to suspended biomass in R1 was 7.7 in absence of SCN–. This ratio increased to 13.5 with addition of 110 mg/L thiocyanate as biomass in suspension decreased from 1400 mg/L to 700 mg/L (decreased by almost 50%) and then this ratio decreased to 10.6–10.2 at feed thiocyanate more than 200 mg/L. This indicates higher amount of biomass was in attached condition to sponge cube than the suspended biomass concentration through out the study.
44–70% of influent SCN– [Table 4.2 (a)]. Figure 4.4 shows that with increase in SCN– loading in R2, SCN– removal rate in R2 increased almost linearly form 0.024 to 0.166 g/L.day with a slope of 0.61. Figure 4.3 (a) shows that almost 21–32% of feed SCN– was removed in R2, which initially increased with increase in feed SCN– up to 450 mg/L and with further increase in feed SCN– it decreased slightly to 27%. In published literatures reports on degradation of SCN– in anoxic environment is quite contradictory.
Kim et al. (2008a), reported phenol and COD removal of 69–32% and 42–31% at a phenol loading rate of 0.06–0.25 g/L.day and COD loading of 1.03–1.86 g/L.day, respectively without any SCN– removal in presence of 82–128 mg/L NH4+–N in the anoxic unit of a pre–denitrification system. Shieh and Richards (1988) too reported no thiocyanate removal in anoxic reactor. However, Andreoni et al. (1988) first reported SCN– degradation by mixed denitrifying cultures with end product as sulfate. Sorokin et al. (2004) also reported SCN– removal in anoxic reactor by sulfur oxidizing autotrophic bacteria, Thialkalivibrio thiocyanodenitrificans, where SCN– was used as an electron donor with nitrate/nitrite as electron acceptors and end products were sulfate and ammonia (Eq.2.2d). Grigor’eva et al.
(2009) also observed ammonium and sulfate as end products of thiocyanate degradation under both aerobic and denitrifying condition. However, these reported studies were carried out without any phenol in medium. Present investigation shows that SCN− can be efficiently biodegraded by mixed denitrifying cultures in presence of phenol. Influent and effluent sulfate values in R2 are presented in Table 4.2 (a).
Figure 4.4 Effect of thiocyanate loading on removal rate in R2
y = 0.6102x R2 = 0.9707
0.00 0.05 0.10 0.15 0.20
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35
Thiocyanate loading (g/L.day) Thiocyanate removal rate (g/L.day)
In R2, some sulfate entered as influent from R3 with the recycle (95–365 mg/L). Table 4.2 (a) shows that 140–600 mg/L sulfate got released in R2 effluent with 45 –235 mg/L of sulfate generation in R2. Generation of sulphate in R2 was possible only from SCN– degradation. Based on the generation of byproducts, the stoichiometric equation of SCN– degradation in anoxic reactor is proposed below (Sorokin et al. 2004):
- - + - 2-
3 2 3 4 3 2 2
SCN + 1.6NO -N + 0.2H O+ 1.6H + HCO SO + NH + 2CO + 0.8N (Eq.4.3.a) It was calculated that Gibb’s free energy change at standard state (∆Go(aq)) for this reaction was (–)824.01 kJ, suggesting feasibility of the reaction at standard state. The stoichiometric equation for SCN– removal in aerobic environment is as equation 4.3 (b) (Staib and Lant, 2007) with ∆Go(aq) of (–)828.34 kJ, suggesting aerobic SCN– degradation is thermodynamically more feasible than anoxic one.
- 2- +
2 2 4 4 2
SCN + 2O + 2H OSO + NH + CO (Eq.4.3.b) Table 4.2 (a): Average performance of anoxic CMBR (R2) at feed SCN–
concentration variation
SCN– Phenol COD Sulfate
S0 Se Rem S0 Se Rem S0 Se Rem S0 Se Gen Th
SO42–
Err
– – – 644 0 100 2005 620
(21)
69.0 – – – – –
54 (0.1)
30 (3.1)
44.3 850 (4.7)
344 (18)
59.5 2682 950 (10)
65.0 95 (0.8)
140 (3.5)
45 39.6 7.2
96 (2.3)
45 (5.7)
52.6 929 (16.8)
410 (27)
55.9 3127 947 (43)
70.0 136 (8)
210 (12)
74 83.2 –9.2
208 (3.7)
61 (8.5)
70.6 1069 (25.9)
450 (23)
57.9 3326 915 (30)
72.5 353 (19)
590 (27)
237 242.5 –5.3
288 (0.8)
122 (12)
57.6 1093 (19.3)
468 (12)
57.2 3980 1317 (30)
67.0 365 (13)
600 (23)
235 274.0 – 38.3 S0: Influent (mg/L), Se: Effluent (mg/L), Rem: Removal (%) Gen: Generation (mg/L);
Th. SO42–: Theoretical sulfate generated (1.65x SCN– removed) mg/L;
Err: Error, experimental value–theoretical value, (mg/L) Numbers in parenthesis indicate standard deviation values.
Maximum error in sulfate generation was observed to be (–) 38 mg/L compared to the theoretical sulfate generation at maximum thiocyanate loading of 0.28 g/L.day. Balance in thiocyanate degradation and sulfate generation is rarely reported. Mahmood et al. (2008) observed less generation of sulfate during oxidation of sulfide in anoxic reactor at high pH (> 8.0) and higher sulfide loading though sulfide removal was steady. Buisman et al.
(1990) also reported accumulation of other sulfur compound such as thiosulfate, polysulfides in anoxic environment in same condition of high pH and high sulfide loading.
In the present study lower generation of sulfate than that of theoretical value at higher loading of thiocyanate might be due to conversion of sulfate to other undetected sulfur compound like polysulfides etc.
Influent phenol concentrations to R2 were 640, 851, 929, 1069 and 1093 mg/L [Table 4.2 (a)] with loading rates of 0.64, 0.851, 0.929, 1.069 and 1.093 g/L.day, respectively.
Reactor R2 removed 56–60% of influent phenol in presence of 54–288 mg/L SCN–. Figure 4.5 shows that phenol removal rate in R2 decreased little due to presence of SCN– in influent of R2.
Figure 4.5 Effect of thiocyanate loading on phenol and COD removal rates in R2
0.0 0.5 1.0 1.5 2.0 2.5 3.0
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35
Thiocyanate loading (g/L.day) COD/Phenol rmeoval rate (g/L.day)
COD Phenol
The average influent COD to R2 were 2005, 2682, 3127, 3326 and 3980 mg/L. COD loading rates were 2.05, 2.68, 3.12, 3.32 and 3.98 g/L.day. R2 removed 64–72% of influent COD showing 23–35% of total feed COD removal. It was observed that with increase in SCN– loading, COD removal rate in R2 increased, suggesting that the inhibitory affect of SCN– on phenol and COD removal was absent in R2 unlike upstream reactor R1. Figure
4.3 shows that fractional phenol and COD removal by R2 was in the range of 20–25% and 25–35% respectively, irrespective of feed SCN–.
Influent ammonia to R2 ranged from 251–338 mg/L [Table 4.2(b)] and the loadings were 0.251, 0.281, 0.292, 0.314 and 0.338 g/L.day. In R2, some amount of NH4+–N was expected to be generated due to degradation of SCN– (0.24 g of NH4+–N/g SCN– removed) and 7.5–2.6% of NH4+–N removal occurred which might be due to incorporation in the synthesis of biomass.
Table 4.2 (b): Average performance of anoxic CMBR (R2) at SCN– concentration variation
NOx––N SCN–
NO3––N NO2––N
NH4+–N Feed
SCN–
S0 S0# Se S0 Se
Rem
S0A Se Rem
COD:
N remψ
CODB TVSC
0 – 727 316
(6)
0 0 56.6 251 220 12.35 3.36 15.0 10580
110 54
(0.1)
617 (9)
47 (2)
2 0 92.4 281
(8.8) 260 (7.6)
7.42 3.03 5.6 9125
200 96
(2.3)
627 (3)
45 (1)
3.5 0 92.8 292 (1.8)
267 (7.5)
8.45 3.76 23.9 9432
450 208 (3.7)
624 (6)
120 (9)
29 0 81.6 314
(11) 270 (4.2)
14.10 4.91 41.7 10420
600 288 (0.8)
612 (8)
129 (9)
50 0 80.5 338
(5.5) 330 (12)
2.60 5.60 49.8 11355
S0: Influent (mg/L), Se: Effluent (mg/L), Rem: Removal (%),
# In R2 influent 500 mg/L of NO3––N was added externally;
A Influent NH4+–N of R2 = {Effluent NH4+–N of (R1+R3)/2 + 0.24x (SCN– removed in R2)}.
ψ: as per equation (4.5); CODB: COD fraction (%) for biomass;
CTVS: Biomass as Total volatile solids in sponge + suspension (mg/L);
Numbers in parenthesis indicate standard deviation values.
With introduction of SCN– in R2, total volatile solids (TVS) initially found to decrease from 10500 mg/L to 9125 mg/L. However it increased to 11300 mg/L while high concentration of SCN– was present in the influent to R2. In absence of SCN– in R2, high
ratio of attached biomass to suspended biomass was observed (14:1). With increase in influent SCN– the ratio continuously decreased. Up to influent SCN– concentration 96 mg/L the ratio was 9.2 and it decreased to 4.1 and 3.6 at further increase in influent SCN– concentration to 208 and 288 mg/L, respectively. Sludge rising in R2 was observed regularly due to production of nitrogen gas inside the reactor. The entrapped gas was released by inserting a thick bamboo stick to the interior of the reactor and again closing R2 at the upper end.
In the present study, NO3––N concentration in recycle (effluent of R3) was inadequate, so additional nitrate (1000 mg/L NO3––N) was supplied externally in recycled effluent of R3 in order to maintain strict anoxic condition in R2. Previous literatures reported commencement of methanogenic condition in anoxic reactor, after depletion of nitrate/nitrite (Karim and Gupta, 2003). Influent NO3–
–N to R2 were 727, 617, 627, 624 and 612 mg/L with volumetric loadings of 0.612– 0.727 g/L.day. From Table 4.2 (b), it can be seen that with increase in feed SCN–, influent NO3–
–N to R2 decreased, whereas influent NO2–
–N increased. In R2, though complete removal of NO2–
–N was achieved, removal of NO3–
–N was incomplete, ensuring anoxic condition in R2. In absence of SCN– NOx–N (NO3–
–N + NO2–
–N) removal was only 56% which further increase to 80–92%
with addition of SCN– in R2. Equation 4.3 shows that 0.38 g NO3–
–N was required for each g of SCN– oxidation in anoxic environment. Theoretically, for each gram of phenol, required NO3–
–N is 0.83 g [equation 2.1 (b)]. Stochiometrically 2.86 g and 1.71 g COD are consumed for removal of one g of NO3–
–N and NO2–
–N, respectively [equation 4.4 (a–c)]
(Sarfaraz et al. 2004). However, the actual ratio may be higher than this stoichiometric value, since some amount of COD is also consumed for synthesis of biomass.
+ -
2 2
O + 4H + 4e 2H O (Eq.4.4.a)
- + -
3 2 2
NO + 6H + 5e 0.5N +2H O (Eq.4.4.b)
- + -
2 2 2
NO + 4H +3e 0.5N +2H O (Eq.4.4.c)
The ratio of COD consumed for unit amount of nitrate was calculated using equation 4.5, with the correction of NO3–
–N consumed for SCN–oxidation (0.38 mg/mg SCN–), COD consumed for nitrite reduction (1.71 mg/mg nitrite) and COD generated from SCN–(1.1 mg/mg SCN–.
- - - -
2 2
- - - -
3 3
rem
COD COD SCN SCN NO -N NO -N
NO -N NO -N SCN SCN
(Influent - Effluent )-1.1x (Influent - Effluent -1.71x (Influent (Influent - Effluent ) -0.38x (Influent - Effluent ) COD:N
) - Effluent )
=
(Eq. 4.5)
Table 4.2 (b) shows that the COD/NO3—Nremratio was 3.03–5.6 in R2 during variation of feed SCN– study. When the theoretical value of 2.86 was divided with this value of COD/Nrem, (3.0–5.6), it provided 0.50–0.94. This suggest that 50 to 94% COD was utilized for reduction of NO3––N and residual 6–50% COD was consumed for synthesis of biomass in R2. With higher amount of COD removed, COD fraction available for biomass synthesis (CODB) in R2 increased. Considering 1.42 as COD of the biomass (C5H7NO2), the observed yield of biomass in R2 was 0.04–0.35 (Metcalf and Eddy, 2003). Tuesseau–
Vuillemin et al. (2003) reported bacterial growth yield greatly varies from 0.2 to 0.6 and mostly considered to be 0.4. Chakraborty and Veeramani (2006) reported bacterial yield of 0.22–0.24.
Another study was conducted to find out the necessity of nitrate in thiocyanate degradation and therefore separate six 5 L aspirator bottle, acclimatized bacteria similar to R2 and B2 operated at a draw and fill mode with 16% sponge cube were applied in presence of phenol. Nitrate concentration was varied as 0, 50, 75, 100, 200 and 350 mg/L along with thiocyanate and phenol concentration 40 and 200 mg/L, respectively and reactor HRT maintained at 5 days. The variation of influent NO3––N showed the most profound effect on SCN− removal [Table 4.2 (c)]. With increase in feed NO3––N concentration from 50 to 350 mg/L, removal of SCN– in the reactor increased from 67% to 95% from influent of 40 mg/L which was only 5% in absence of NO3––N. Sulphate was observed in the effluent and increased from 44 to 60 mg/L, with increase in influent NO3––N and was near to the theoretical sulphate generation from thiocyanate degradation. Effluent NH4+–N also increased to 11-14 mg/L from influent of 10 mg/L. With increase in influent NO3––N, denitrification efficiency increased from 88% to 92%, up to feed NO3––N of 100 mg/L and beyond this denitrification efficiency decreased probably due to less availability of electron donor, since phenol and SCN–were kept constants at 200 and 40 mg/L, respectively. The study shows that thiocyanate degradation is feasible in anoxic environment and nitrate is essential.
Table 4 2 (c): Performance of 5L anoxic batch reactor at varying NO3––N concentration
NO3––N COD Phenol SCN– NH4+–
N
Sulfate
S0 Se Rem S0 Se Rem Se Rem Se Rem Se
TVS (mg/L)
Se Th SO42–
Err
50 6 (2.1) 88.0 700 242.0 (15)
65.43 79.0 (1.6)
60.5 13.00 (1.2)
67.5 12.0 (0)
5211 (440)
44.0 (2)
44.5 –0.5
75 6 (3) 92.0 710 152.0 (30)
78.59 8.8 (1.7)
95.6 9.35 (3)
76.6 13.0 (10)
7202 (555)
45.0 (2)
50.5 –4.5
100 8 (0.9) 92.0 720 138.0 (22)
80.83 0.6 (0) 99.7 6.35 (1.3)
84.1 14.0 (0)
8220 (548)
50.0 (3)
55.5 –4.5
200 38 (2) 81.0 720 135.0 (17)
81.25 1.3 (0) 99.3 1.90 (1)
95.2 12.0 (0)
9432 (600)
58.0 (2.3)
62.8 –3.2
350 127.0 (3)
63.7 720 120.0 (22)
83.33 0.8 (0) 99.6 1.65 (0)
95.4 12.0 (0)
9440 (600)
60.0 (2.5)
63.2 –3.2
0 0 NA 700 550.0
(20)
21.43 120.0 (1.1)
40.0 38.00 (1)
5.0 11.0 (0)
3930 (580)
– – –
S0: Influent (mg/L); Se: Effluent (mg/L); Rem: Removal (%) Err: Error (mg/L);
Influent phenol, SCN–and NH4+–N were constant at 200, 40 and 10 mg/L, respectively.
Th SO42–: Theoretical sulfate generation (mg/L); [1.65* (SCN– removed (mg/L)]