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Sulfate-reducing bacteria
Sulfate-reducing bacteria
Sulfate-reducing bacteria (SRB) form one group of sulfate reducing prokaryotes. Main genus is Desulfovibrio. Desulfovibrio desulfuricans is often used to immobilize dissolved heavy metals as metallic sulfides.
Beijerinck[1] showed in 1895 that living matter could reduce sulphate to sulphide in sediments under anaerobic conditions. Although many bacteria can produce sulphide, only a few do so at a sufficient rate for application in high-rate processes. These rapid sulphide-generating bacteria are able to conserve energy by the reduction of sulfur oxyanions[2], and they are generally termed sulphate-reducing bacteria (SRB). A typical overall conversion equation is (neglecting the small amount of organic material required to produce biomass):
- SO42- + CH3COOH + 2 H+ → HS- + 2 HCO3- + 3 H+ (1)
Eight electrons are transferred from the energy source acetic acid to the electron acceptor sulphate in order to produce sulphide. The reaction equation shows that in the same process also alkalinity is produced. This leads to an increase in the pH of the water, often to a near neutral value.
Typically, a certain amount of metals is present together with the sulfate. These metals will react with the dissolved sulfide to form highly insoluble metals sulfides.
- HS- + Me2+ → MeS + H+ (2)
Me2+ can for example be copper, zinc etc. Andean region.
The effluents produced in sulphide ore mines, defined as acid mine drainage (AMD), also contain large amounts of heavy metals. Mining and industrial drainage containing sulphate and heavy metal negatively affects terrestrial and aquatic ecosystems in several countries around the world.Sulphate-reducing bacteria (SRB) can be used to biologically treat sulphate-rich wastewater. SRB comprise several groups of bacteria that reduce sulphate to sulphide and produce carbonate which increase the pH. In AMD treatment processes this chemically stabilizes the toxic metal ions as solid metal sulphides[3].
Hydrogen sulphide
The reduction product of reaction I, hydrogen sulphide, is a volatile gas. The form in which sulphide occurs depends on the pH:
- H2S → HS- + H+ → S2- + 2H- (3)
HS- and S2-which occur at neutral and high pH respectively are both water soluble. H2S is the predominant form at low pH (<6)[4][5].
Sulphide is distributed over the gas phase (g) and the liquid phase (l) according to:
- [H2S]l =α*[H2S]g (mol/m3) (4)
α is a dimensionless distribution coefficient. The unionised H2S concentration also depends on the temperature. Sulphide is highly reactive, corrosive and toxic to microorganisms [6]. The toxicity increases at low pH while only the un-ionised hydrogen sulphide form is able to permeate through the cell membrane. H2S affects the intracellular pH of the microorganism and impedes its metabolism [7][8].
Classification of SRB
SRB are obligate anaerobes and members of a heterogeneous group of eubacteria and archaebacteria which are able to carry out dissimilatory sulphate reduction (Colleran et al.,1995; Hansen, 1994). The SRB can be subdivided into two groups depending on their oxidative capability: the genera that completely oxidise the organic substrate to CO2, and the bacteria that oxidise the organic compound incompletely usually with acetate as an end product. The species able to completely oxidise organic carbon sources mainly prefers fatty acids, lactate and succinate as energy sources. Incomplete oxidation is due to the absence of a mechanism for acetyl-Co-A oxidation. Such bacteria generally prefer simple substrates such as hydrogen, lactate and primary alcohols (Alvarez, 2005; Kolmert, 1999; Vallero, 2003).
SRB can survive in a wide range of pH conditions but commonly have a pH optimum for growth between pH 5-9 (Jong et al., 2006). SRB populations have been obtained at temperatures ranging from the [psychrophilic]] to the hyperthermophilic range (Kolmert, 1999).
Extremophilic SRB
Regarding the applications for biological treatment processes, the significance of some extremophilic bacteria should be emphasized. Among the diversity of sulphate-reducing prokaryotes the acidophilic, thermophilic and psychrotolerant bacteria are extremophiles that could improve the performance of existing treatment systems.
Acidophilic SRB
During mining activities oxygen is introduced into deep geological environments and cause chemical and biological oxidation processes. Sulphate and hydrogen ions are produced which lower pH significantly (Kolmert, 1999; Madigan et al., 2000). The pH is generally between 2 and 4 and commonly less than 3. Current biological acid mine drainage treatment systems mainly use neutrophilic SRB, highly sensitive to acidic water; resulting in few successful applications (Jong et al., 2006). To run the system “off line” is a method to circumvent this problem. The SRB grow in an isolated neutral tank where hydrogen sulphide is produced and the effluent is transferred to a second reactor. This tank contains the contaminated water which results in precipitation of metal sulphide. Acidophilic or acido-tolerant bacteria are able to grow in direct contact with the acidic liquid in a single reactor tank. This could simplify the system and be a less expensive solution to the two tanks treatment system existing today (Kimura et al., 2006; Kolmert et al., 2001).
Thermophilic SRB
Wastewater containing sulphur compounds is generated by several industrial processes. Examples of industries contributing to imbalances in the natural sulphur cycle are those using sulphuric acid or sulphate-rich feed stocks, such as the food and fermentation industry. Some industrial wastewaters are discharged at high temperatures of 50 to 70ºC and even above 90ºC. The use of thermophilic SRB to treat such wastewater holds some advantages and may be an attractive alternative to treat the discharge mesophilically. It eliminates the cooling of the process water and allows direct use of the treated water without additional re-heating. Besides, the termophilic systems produce less sludge and are capable of treating higher organic loading rates with feasible removal efficiency (Vallero, 2003; Pender et al., 2004).
Cold-adapted SRB
Treatment of AMD and industrial wastewater functional in low temperatures are of potential interest in countries with cold environments. It could be realized by the use of psychrotolerant sulphate-reducing prokaryotes. Psychrophilic SRB generally have a growth optimum temperature of 18ºC while the optimum for sulphate reduction is 28ºC. However, bacteria reducing sulphate below 4°C have been identified. SRB are sometimes less active in low temperatures and the lower reaction rates of the process could be compensated by an increased number of bacteria (Knoblauch et al., 1999; Sahm et al., 1999).
Expression Of Analytical Results
Water analysis involves the detection of minute amounts of a variety of substances. The expression of results in percentage would require the use of cumbersome figures. For this reason, the results of a water analysis are usually expressed in parts per million (ppm) instead of percentage. One part per million equals one ten-thousandth of one percent (0.0001%), or one part (by weight) in a million parts-for example, 1 oz in 1,000,000 oz of water, or 1 lb in 1,000,000 lb of water. It makes no difference what units are used as long as both weights are expressed in the same units.
When elements are present in minute or trace quantities, the use of parts per million results in small decimal values. Therefore, it is more convenient to use parts per billion (ppb) in these cases. One part per billion is equal to one-thousandth of one part per million (0.001 ppm). For example, in studies of steam purity using a specific ion electrode to measure sodium content, values as low as 0.001 ppm are not uncommon. This is more conveniently reported as 1.0 ppb.
In recent times, the convention for reporting analytical results has been shifting toward the use of milligrams per liter (mg/L) as a replacement for parts per million and micrograms per liter (µg/L) as a replacement for parts per billion.
Test procedures and calculations of results are based on the milliliter (mL) rather than the more common cubic centimeter (cc or cm3). The distinction between the two terms is very slight. By definition, a milliliter is the volume occupied by 1 g of water at 4°C, whereas a cubic centimeter is the volume enclosed within a cube 1 cm on each edge (1 mL = 1.000028 cm3).
The milligrams per liter (mg/L) convention is closely related to parts per million (ppm). This relationship is given by:
ppm x solution density = mg/L
Thus, if the solution density is close or equal to 1, then ppm = mg/L. This is normally the case in dilute, aqueous solutions of the type typically found in industrial water systems. Control testing is usually conducted without measurement of a solution's density. For common water samples, this poses no great inaccuracy, because the density of the sample is approximately 1. Milligrams per liter (mg/L) and parts per million (ppm) begin to diverge as the solution density varies from 1. Examples of this are a dense sludge from a clarifier underflow (density greater than 1) or closed cooling system water with high concentrations of organic compounds (density less than 1). All of the analytical methods discussed in this text contain calculations required to obtain the results in milligrams or micrograms per liter.
In reporting water analyses on an ion basis, results are also expressed in equivalents per million (epm). Closely allied to the use of parts per million, this approach reduces all constituents to a common denominator-the chemical equivalent weight.
The use of equivalents per million is not recommended for normal plant control. Parts per million is a simpler form of expressing results and is accepted as the common standard basis of reporting a water analysis. However, whenever extensive calculations must be performed, the use of equivalents per million greatly simplifies the mathematics, because all constituents are on a chemical equivalent weight basis. The remainder of this section provides a discussion of parts per million and equivalents per million for those who desire a working knowledge of these methods of expression for purposes of calculations.
The units of ppm and epm are commonly combined in normal reporting of water analyses, and many different constituents are frequently reported on a common unit weight basis. For example, calcium (equivalent weight 20.0) is reported in terms of "calcium as CaCO3" (equivalent weight 50.0). The test for calcium is calibrated in terms of CaCO3, so the conversion factor 2.5 (50/20) is not needed. Hardness, magnesium, alkalinity, and free mineral acid are often reported in terms of CaCO3; the value reported is the weight of CaCO3 that is chemically equivalent to the amount of material present. Among these substances, ionic balances may be calculated. When constituents are of the same unit weight basis, they can be added or subtracted directly. For example, ppm total hardness as CaCO3 minus ppm calcium as CaCO3 equals ppm magnesium as CaCO3. However, ppm magnesium as Mg2+ equals 12.2 (magnesium equivalent weight) divided by 50.0 (CaCO3 equivalent weight) times the ppm magnesium as CaCO3.
In every case, it is necessary to define the unit weight basis of the results-"ppm alkalinity as CaCO3" or "ppm sulfate as SO42- " or "ppm silica as SiO2". Where the unit weight basis is different, calculations must be based on the use of chemical equations.
The following rules outline where epm can be used and where ppm must be used. In general, either may be used where an exact chemical formula is known. When such knowledge is lacking, ppm must be used.
- The concentration of all dissolved salts of the individually determined ions must be in ppm.
- Two or more ions of similar properties whose joint effect is measured by a single determination (e.g., total hardness, acidity, or alkalinity) may be reported in either ppm or epm.
- The concentration of undissolved or suspended solids should be reported in ppm only.
- The concentration of organic matter should be reported in ppm only.
- The concentration of dissolved solids (by evaporation) should be expressed as ppm only.
- Total dissolved solids by calculation may be expressed in either ppm or epm.
- Concentration of individual gases dissolved in water should be reported in ppm. The total concentration of each gas when combined in water may be calculated to its respective ionic concentration in either ppm or epm.
CALCULATION OF TOTAL DISSOLVED SOLIDS BY EPM
Starting with a reasonably complete water analysis, total dissolved solids may be calculated as epm. In a complete water analysis, the negative ion epm should equal the positive ion epm. Where there is an excess of negative ion epm, the remaining positive ion epm is likely to be sodium or potassium (or both). For the sake of convenience, it is generally assumed to be sodium. Where there is an excess of positive epm, the remaining negative epm usually is assumed to be nitrate.
To calculate dissolved solids, convert the various constituents from ppm to epm and total the various cations (positively charged ions) and anions (negative ions). The cations should equal the anions. If not, add either sodium (plus) or nitrate (minus) ions to balance the columns. Convert each component ionic epm to ppm and total to obtain ppm dissolved solids. For example, to convert 150 ppm calcium as CaCO3 to epm (Table 40-1) divide by 50 (the equivalent weight of calcium carbonate) and obtain 3.0 epm. To convert 96 ppm sulfate as SO42- to epm, divide by 48 (the equivalent weight of sulfate) and obtain 2.0 epm. After balancing the cations and anions by adding sodium, convert to ionic ppm by multiplying the epm by the particular ionic equivalent of weight. For example, to convert 3.0 epm calcium to ppm calcium as Ca2+, multiply by 20 (the equivalent weight of calcium) and obtain 60 ppm calcium as Ca2+. To obtain the ppm dissolved solids, total the ppm of the individual ions.
Table 40-2: Conversion Table
|
Formula |
Number of equivalents |
Equivalent Weight |
|
POSITIVE IONS |
||||
Aluminum |
Al+3 |
3 |
9.0 |
|
Ammonium |
NH4+ |
1 |
18.0 |
|
Calcium |
Ca2+ |
2 |
20.0 |
|
Copper |
Cu2+ |
2 |
31.8 |
|
Hydrogen |
H+ |
1 |
1.0 |
|
Ferrous Ion |
Fe2+ |
2 |
27.9 |
|
Ferric Ion |
Fe3+ |
3 |
18.6 |
|
Magnesium |
Mg2+ |
2 |
12.2 |
|
Manganese |
Mn2+ |
2 |
27.5 |
|
Potassium |
K+ |
1 |
39.1 |
|
Sodium |
Na+ |
1 |
23.0 |
|
NEGATIVE IONS |
||||
Bicarbonate |
HCO3- |
1 |
61.0 |
|
Carbonate |
CO32- |
2 |
30.0 |
|
Chloride |
Cl- |
1 |
35.5 |
|
Fluoride |
F- |
1 |
19.0 |
|
Iodide |
I- |
1 |
126.9 |
|
Hydroxide |
OH- |
1 |
17.0 |
|
Nitrate |
NO3- |
1 |
62.0 |
|
Phosphate (tribasic) |
PO43- |
3 |
31.7 |
|
Phosphate (dibasic) |
HPO42- |
2 |
48.0 |
|
Phosphate (monobasic) |
H2PO4- |
1 |
97.0 |
|
Sulfate |
SO42- |
2 |
48.0 |
|
Bisulfate |
HSO4- |
1 |
97.1 |
|
Sulfite |
SO32- |
2 |
40.0 |
|
Bisulfite |
HSO3- |
1 |
81.1 |
|
Sulfide |
S2- |
2 |
16.0 |
|
COMPOUNDS |
||||
Alum |
Al2(SO4)3 18H2O |
6 |
111.0 |
|
Aluminum Sulfate (anhydrous) |
Al2(SO4)3 |
6 |
57.0 |
|
Aluminum Hydroxide |
AI(OH)3 |
3 |
26.0 |
|
Aluminum Oxide |
Al2O3 |
6 |
17.0 |
|
Ammonia |
NH3 |
1 |
17.0 |
|
Sodium Aluminate |
Na2AI2O4 |
6 |
27.3 |
|
Calcium Bicarbonate |
Ca(HCO3)2 |
2 |
81.1 |
|
Calcium Carbonate |
CaCO3 |
2 |
50.0 |
|
Calcium Chloride |
CaCl2 |
2 |
55.5 |
|
Calcium Hydroxide |
Ca(OH)2 |
2 |
37.0 |
|
Calcium Oxide |
CaO |
2 |
28.0 |
|
Calcium Sulfate (anhydrous) |
CaSO4 |
2 |
68.1 |
|
Calcium Sulfate (gypsum) |
CaSO4 2H2O |
2 |
86.1 |
|
Calcium Phosphate |
Ca3(PO4)2 |
6 |
51.7 |
|
Carbon Dioxide |
CO2 |
2 |
22.0 |
|
Chlorine |
Cl2 |
2 |
35.5 |
|
Ferrous Sulfate (anhydrous) |
FeSO4 |
2 |
76.0 |
|
Ferric Sulfate |
Fe2(SO4)3 |
6 |
66.6 |
|
Magnesium Oxide |
MgO |
2 |
20.2 |
|
Magnesium Bicarbonate |
Mg(HCO3)2 |
2 |
73.2 |
|
Magnesium Carbonate |
MgCO3 |
2 |
42.2 |
|
Magnesium Chloride |
MgCl2 |
2 |
47.6 |
|
Magnesium Hydroxide |
Mg(OH)2 |
2 |
29.2 |
|
Magnesium Phosphate |
Mg3(PO4)2 |
6 |
43.8 |
|
Magnesium Sulfate (anhydrous) |
MgSO4 |
2 |
60.2 |
|
Magnesium Sulfate (Epsom Salts) |
MgSO4. 7H2O |
2 |
123.2 |
|
Manganese Hydroxide |
Mn(OH)2 |
2 |
44.5 |
|
Silica |
SiO2 |
2 |
30.0 |
|
Sodium Bicarbonate |
NaHCO3 |
1 |
84.0 |
|
Sodium Carbonate |
Na2CO3 |
2 |
53.0 |
|
Sodium Chloride |
NaCl |
1 |
58.4 |
|
Sodium Hydroxide |
NAOH |
1 |
40.0 |
|
Sodium Nitrate |
NaNO3 |
1 |
85.0 |
|
Trisodium Phosphate |
Na3PO4. 12H20 |
3 |
126.7 |
|
Trisodium Phosphate (anhydrous) |
Na3PO4 |
3 |
54.7 |
|
Disodium Phosphate |
Na2HPO4. 12H2O |
2 |
179.1 |
|
Disodium Phosphate (anhydrous) |
Na2HPO4 |
2 |
71.0 |
|
Monosodium Phosphate |
NaH2PO4. H2O |
1 |
138.0 |
|
Monosodium Phosphate (anhydrous) |
NaH2PO4 |
1 |
120.0 |
|
Sodium Silicate |
Na2SiO3 |
2 |
61.0 |
|
Sulfuric Acid |
H2SO4 |
2 |
49.0 |
|
Sodium Metaphosphate |
NaPO3 |
1 |
102.0 |
|
Sodium Sulfate |
Na2SO4 |
2 |
71.0 |
|
Sodium Sulfite |
Na2SO3 |
2 |
63.0 |