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Time COD Cells 02 uptake COD cf cells A COD A Cells A COD cells Calculated substrate recovery (per cent)t
(mg/liter) (mg/liter) (mg/liter) (mg/liter) (mg/liter) (mg/liter) (mg/liter)
hr
0.00 544 450 585
0.5 450 485 13 626 94 35 41 53.5 65 66.5 57.5
1.5 292 573 45 730 252 123 145 70 84 87 75.5
2.0 231 629 64 810 313 179 225 81 98 102 93
2.5 143 715 84 922 401 265 337 91 110.5 114.5 105
3.0 72 755 101 985 472 305 400 90 109 113 106
3.5 26 764 116 960 518 314 375 87 105 108 95
4.0 20 774 129 970 524 324 385 90.5 109 112 98
4.5 20 777 138 1,020 524 327 435 93 111 115 109
5.0 15 774 144 977 529 324 392 92.5 110.5 114 101
*
Intermediate-age cells, acclimated to glucose.
t (I) Materials balance, weight calculation. (II) Materials balance, carbon calculation. (III) Energy balance, based on empirical form-
using old cells, recoveries were higher for all four methods use in waste treatment research. With two sets of data,
than in the previous experiment. The numerical order of balances were calculated by four different methods; the
percentages recovered was the same in both cases; i.e., recommended method yielded better average recoveries
the methods may be arranged, for both experiments, in than any of the others. The proposed balance technique
the same order according to decreasing percentage of would further appear to have a much sounder technical
substrate recovered: III > II > IV > I. Overall average basis than the other methods, since it does not require the
per cent recoveries for the two experiments were: method use of an empirical formula or average carbon content
I, 92.6; method II, 112.2; method III, 116.0; Method IV, for activated sludge nor does it require that the chemical
102.3. components of the waste be known. Thus, the recom-
Sample calculations for substrate recovery. Method I: mended procedure is more widely applicable, since it may
Conversion factor for COD or 02 to weight of substrate = be employed with equal facility in studies on whole wastes
0.94, since 6 moles (192 g) of 02 are required for complete or on synthetic wastes with known composition. If the
oxidation of 1 mole (180 g) of glucose composition of the waste is known, method I may be
= 144(0.94) + 324 preferable in some cases since it yields adequate recovery
Recovery 52(.9) 0.925 =
data, and the analyses required are much less time-
consuming than for method IV.
M\ethod II: Assuming moles of CO2 = moles of 02 for In using cell COD to calculate an energy balance, it is
glucose, then weight of CO2 = weight of 02 X 432, not recommended that an average value for cell COD
CO2 carbon = 1244 X weight of C02, cell carbon = be employed to convert sludge weight to an equivalent
weight of cells X 0.51, weight of substrate used = COD X COD. Whereas in some experiments the COD of the cells
0.94, weight of substrate carbon used = weight of substrate remained quite constant throughout the experimental
used X 72/80O period, in other experiments the COD of the cells varied
over a broad range. In the latter cases, a considerable
Recovery = 144(1.375) (0.273) + 324(0.51) 1 105 error would have been introduced by use of an average
529(0.94)(0.4) COD value even though the average might have been
M\Iethod III: Calculated COD of cells = 1.414 X weight experimentally determined for that particular sludge.
of cells Use of a general overall average COD value for cells
144 + 1.414(324) might introduce even greater error. Our data indicate
Recovery = 59
529 =1.14 the possibility that both cell age and the particular sub-
strate used may affect the COD of the cells, although not
Method IV: enough data are available to allow definite conclusions to
be made in this respect.
Recovery 4439= 1. 01 The most valid objection to the use of the proposed
5-29
method of determining substrate balances lies in the
DISCUSSION difficulty of demonstrating complete oxidation of cell
From the results herein presented, it is concluded that material by the COD method. It is not possible to state
an energy balance based on determinations of the COD of that 100 % oxidation of cellular material was achieved;
the culture filtrate, the COD of the cells produced, and however, the standard COD test does apparently oxidize
the oxygen utilized by the cells can be recommended for all the cell material which acid dichromate is capable of
260 GAUDY, BHATLA, AND GAUDY APPL. MICROBIOL.
oxidizing, since increasing the reflux time to as long GAUDY, A. F., JR. 1962. Colorimetric determination of protein and
as 24 hr resulted in no further oxidaflon of the cells. COD carbohydrate. Ind. Water Wastes 7:17-22.
GAUDY, A. F., JR., AND R. S. ENGELBRECHT. 1960. Basic biochemi-
values obtained with replicate samples of different amounts cal considerations during metabolism in growing vs respiring
of cells were also highly reproducible. systems. Proc. Conf. Biol. Waste Treat., 3rd, Manhattan Coll.,
Further support for the use of the COD test for activated New York, N.Y.
sludges is provided by the recent report of Goldstein and GAUDY, A. F., JR., E. T. GAUDY, AND K. KOMOLRIT. 1963a. Multi-
Lokatz (1963) that the COD value of activated sludge component substrate utilization by natural populations and a
provides a fairly accurate measure of its heat of combus- pure culture of Escherichia coli. Appl. Microbiol. 11:157-162.
GAUDY, A. F., JR., K. KOMOLRIT, AND M. N. BHATLA. 1963b. Se-
tion. According to their measurements, a value of 10 g per quential substrate removal in heterogeneous populations. J.
liter of COD corresponds to a heat of combustion of 500 Water Pollution Control Federation 35:903-922.
BTU per gallon. Although this correlation does not GOLDSTEIN, A., AND S. LOKATZ. 1963. Sewage sludge oxidation at
provide absolute assurance that all organic matter in the Chicago. 36th Annual Water Pollution Control Federation
cells is totally oxidized, it does provide evidence that a Conference, Seattle, Wash.
GRADY, L., JR., AND A. W. BUSCH. 1963. BOD progression insoluble
constant proportion of the organic matter in activated substrates. VI. Cell recovery techniques in the TbOD test.
sludge is oxidized in the standard COD test. The fact Proc. 18th Annual Industrial Waste Conference, Purdue Uni-
that in our studies recoveries close to 100 % wer& obtained versity, Lafayette.