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The Chemical Oxygen Demand of

Waters and Biological Materials from
Ponds
a
Claude E. Boyd
a
Department of Fisheries and Allied Aquaculture , Auburn
University Agricultural Experiment Station , Auburn , Alabama ,
36830 , USA
Published online: 09 Jan 2011.

To cite this article: Claude E. Boyd (1973) The Chemical Oxygen Demand of Waters and Biological
Materials from Ponds, Transactions of the American Fisheries Society, 102:3, 606-611, DOI:
10.1577/1548-8659(1973)102<606:TCODOW>2.0.CO;2

To link to this article: http://dx.doi.org/10.1577/1548-8659(1973)102<606:TCODOW>2.0.CO;2

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 The Chemical Oxygen Demand of Waters and
Biological Materials from Ponds
CraUDE E. BOYD
Department o/ Fisheries and Allied Aquaculture, Auburn University
Agricultural Experiment Station, Auburn, Alabama 36830

ABSTRACT

The rate of oxygenconsumption by organismsin pond waters,as measuredby a dark bottle

techniquefor a 24-hr period, increasedwith increasingchemicaloxygendemand (COD). Rate
of oxygenconsumptionincreasedwith temperaturebetween15 and 35 C (Q•ovalueswere 1.5 to
2.0). At the samelevelof COD, oxygenconsumption
wasgreaterin unalteredpondwaterthan
in water in which the plankton was heat-killed and oxygenwas utilized only by bacteria and
other decomposers. COD was positivelycorrelatedwith increasingconcentrations of chlorophyll
in pond waters. The amount'of COD in solubleorganicmatter rangedfrom 20.2 to 95.0% of
the total COD. The percentageof the COD in the solublefraction decreasedwith increasing
quantitiesof phytoplankton.The averageamountsof oxygen(mg oxygen/mgdry wt) required
to completelyoxidizevarioustypesof biologicalmaterialwere; phytoplankton frompurecultures
1.29, macroscopicalgae 1.00, higher aquatic plants0.99, particulatematter from pond waters
0.98, and fish 1.19.
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INTRODUCTION organismsduring a shortperiod (12 to 24 hr)

Ponds that receive applicationsof fertilizer in ponds has not been assessed.The chemical
and fish feed frequentlydevelopdenseblooms oxygen demand (COD) is the amount of
of phytoplankton and contain large concentra- oxygen required to completely oxidize the
tionsof particulateandsolubleorganicmatter. organic matter in a water sample to carbon
dioxide and water. This measurement is ac-
These habitats normally have high densities
of fish and invertebrate animals. Oxygen complishedby the action of strong oxidants
requirements for respiratory processes are which do not distinguishbetweenbiologically
high and considerablephotosynthesisby phy- oxidizableand biologicallyinert organiccom-
toplankton must take place in order to supply pounds. However, if a definite relationship
enough oxygen to sustain aerobic life during existsbetweenoxygen consumptionof plank-
an entire diurnal cycle. High rates of photo- tonic communitiesand COD, the COD values
synthesisare normally observedin fish ponds would be useful in predicting the oxygen
(Hepher, 1962; Boyd, 1972a). There is shal- requirements. The presentstudy was initiated
low thermal stratification in highly productive to determineif a consistentrelationshipexists
pondsand the hypolimnion containshigh con- betweenthe COD of pond waters and the rate
centrations of organic matter. Upwelling of of oxygen consumption by the planktonic
hypolimnetic waters causedby cold air masses, organisms. The contribution of phytoplankton
heavy winds or cold rains may resultin oxygen to the COD of pond waters and the amount of
depletionand fish kills (Swingle, 1968). Fish oxygen required to completely decompose
kills may also occur following suddendie-offs various organismswas also evaluated.
of blooms of phytoplankton. Therefore, it is
MATERIALS AND METHODS
often desirable to obtain measurements of
organic matter concentrations and rates of Water samples were collected from ponds
oxygenconsumptionin fish ponds. on the FisheriesResearchUnit, Auburn Uni-
The biochemicaloxygen demand (BOD) versityduring April and May 1972. Included
gives an estimate of the amount of oxygen were samples from 13 ponds which received
required by bacteria to oxidize the decom- feed applications and samples from 11 fer-
posable organic matter in a water sample. tilized and two unfertilized ponds. Feeds
Standard BOD tests are usually conductedfor contained 7.36% nitrogen and 0.63% phos-
5 days at 20 C. The relationship of standard phorus and were applied six days per week
BOD assays to consumption of oxygen by at the rate of 3% of the body weight of fish
6O6
 BOYD--COD AND ORGANIC MATTER IN PONDS 607

in the ponds. Fertilizer applicationsconsisted catus,Chlamydomonassp., Coelastrummicro-

of monthly additionsof 19.5 kg/ha of triple- porum, and Staurastrum sp. were grown in
superphosphate (46%P205). Samples of water the laboratory as describedby Boyd (1972b).
were collectedin polyethylenebottles from a These algae were filtered onto glass fiber
depth of 10 cm beneath the surface and used filters, dried at 60 C and weighed. Macro-
within 2 hr of collection for preparation of scopic algae samples (Pithophora kewensis,
experiments. Nitella sp.,Rhizocloniumhieroglyphicurn,
and
The rate of oxygen consumptionby com- Spirogyrasp.) and samplesof higher aquatic
munity respiration was determined for 26 plants (Alternanthera philoxeroides,Lemna
samplesby a dark bottle method. Samples minor, Eichhornia crassipes, Myriophyllum
of known dissolvedoxygen content were in- brasiliense,•/allisneria americana, Potamoge-
cubated in duplicate 300-ml BOD bottles at ton sp.,Eloda densa,Eleocharisacicularis,and
30 C for 24 hr in the dark. The decline in Zizaniopsis
miliacea)wereobtainedfromlakes
dissolved oxygen was determined by the and pondsin the vicinity of Auburn,Alabama.
Winkler technique (American Public Health These sampleswere dried at 60 C and pul-
Association, 1960). COD analyses of the verized in a small mill. Samples of fish in-
initial water sampleswere made by the stan- cludedIctalurus catus,I. punctatus,Dorosoma
dard procedure outlined in American Public petense,D. cepedianum,Micropterus salmo-
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HealthAssociation (1960). Glassware usedin ides, Cyprinus carpio, and Lepomis machro-
COD analyseswas washedin H2SO4-Na2Cr207 chirus. Whole fish weredried by lypholization
cleaning solution and glass-distilledwater im- and milled. Samples of particulate matter
mediately prior to use. from pond waters were collectedby filtration
The effect of temperature on oxygen con- of organismsonto tared glass fiber filters as
sumption was determined for samples from describedabove. The total amount of oxygen
four ponds. BOD bottles were filled from required to oxidize samplesof organismswas
samples of known dissolved oxygen content. determined by COD analysis. Samples con-
Duplicate bottles of each water were incubated tained on glassfiber filters or 10-mg samples
in the dark at 15, 20, 25, 30, or 35 C for 24 of other organisms were transferred to the
hr and the decreasein dissolvedoxygen was COD flasks and 20 ml of glass-distilledwater
measured. was added. Digestion reagents were intro-
In anothersetof eightsamples,the plankton duced and the procedure conducted in the
was killed in a water bath heated to 70 C. usual manner. The sampleswere titrated with
Sample bottles were allowed to stand un- 0.25 N ferrous ammonium sulfate. The amount
covered at room temperature for 2 days to of oxygenrequired to oxidize 1 mg of sample
establish bacterial populations. The samples was calculatedfrom the relationshipthat 1 ml
were then usedto prepare a 30 C oxygencon- 0.25 N sodium dichromateconsumedis equiv-
sumption experiment as outlined above. alentto 2 mg of oxygen(Sawyer,1960).
The organicmatter is 26 water sampleswas
partitioned into particulate and soluble frac- RESULTS AND DISCUSSION

tions by filtration through glass fiber filters The COD of 26 pond waters ranged from
(Gelman Type A, 47 mm). All filters were 7.4 to 138.9 mg/liter with an averageof 43.4
previously ashed at 500 C for 2 hr to remove mg/liter. Oxygenutilization by living plank-
organic matter. COD determinations were tonic communities increased proportionally
made on the original samplesand liltrates. with increasesin COD. Rates of oxygen con-
Essentially unialgal samplesof Aphanizo- sumption rangedfrom 0.60to 8.30mg/liter per
menon]los-aquae,Euglenasp., and .4nabaena 24 hr at 30 C (Fig. 1) with an averageof 3.24
circinalis were collected by centrifugation of mg/liter per 24 hr. Water samplescontained
pond waters containing blooms of these several types of phytoplankton communities,
organisms. Sampleswere dried by hypoliza- including fairly unialgal blooms of green or
tion. Pure cultures of other phytoplankters, blue-green algae, blooms containing several
Scenedesmus dimorphus, .4nabaena ]los-aquae, speciesof green algae,and mixed-speciesphy-
Chlorella pyrenoidosa,.dnkistrodesmus ]al- toplanktoncommunitiesof low density. Zoo-
 608 TRANS. AMER. FISH. SOC., 1973, NO. 3
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FmuaE 1.--Relationship between chemical oxygen o TEMPERATURE (* C)

demand and the amount of dissolvedoxygen which
was utilized by organismsin pond waters that were F•cuaz 2.•The effect of temperature on the con-
incubated in the dark for 24 hr at 30 C. sumptionof dissolvedoxygenby organismsin samples
of pond water which was incubated in the dark for
24 hr. The pond designationsrefer to ponds on the
plankton was abundant in most samples,but Fisheries ResearchUnit, Auburn University.
a few samples contained small numbers of
zooplankton. Thirteen of the pondsreceived from 1.4 to 1,066 mg oxygen/m2 per hr. High
daily applicationsof fish feeds,11 received values were associated with benthic commu-
inorganic fertilization, and two receivedno nities comprised of macrovegetation. Values
nutrient additions. Feeds contained 7.36%
for most shallow-water,organically rich sedi-
nitrogen and 0.63% phosphorusand were ments, such as found in many ponds, were
applied6 days per week at the rate of 3% of below 142 mg oxygen/m2 per hr. The con-
thebodyweightof fishin theponds.Fertilizer sumption of oxygen for nine common species
applicationsconsistedoœmonthly additions of freshwater fish at rest ranged from 65 to
of 19.5 kg/ha of triplesuperphosphate
(46% 210 mg/kg per hr at 17 to 20 C (Clausen,
P205). Therefore, the samplesrepresenteda 1936). Oxygenconsumptionby fish increases
wide range of water quality and speciescom- with activity and the consumptionof oxygen
position of the plankton. There probably was at 20 C by five speciesof fish that were forced
a wide variation in the chemical composition
to exercise ranged from 266 to 888 mg/kg
of the particulate and soluble organic matter per hr (Basu, 1959). Oxygen consumption
in the different waters because of the varied
by fish also increasesmarkedly with tempera-
species composition. Since the relationship ture. Values for moderately active Ictalurus
between COD and oxygen consumptionheld catusincreasedfrom 60 mg/kg per hr at 11 C
for such a wide variety of samples, COD in Decemberto 276 mg/kg per hr at 30 C in
appearsto be a useful estimator of community June.1
oxygendemand. However,if COD of waters Oxygen consumptionby plankton commu-
is used in predicting oxygen consumption,
nities increased markedly with temperature
allowancemust be made for oxygenutilization
by fish and benthic communities.
•Shell, E. W. 1965. Fisheries research annual
Pamatmat (1968) summarized data for report, Agricultural Experiment Station, Auburn
various benthic communities which ranged University, Auburn, Alabama.
 BOYD--COD AND ORGANIC MATTER IN PONDS 609

120

•1'
oJ •100
oJ

C9 c=
60
z •40
/e' r =0,909 (P <O,Ot)
• • •o

• • 0.0• + 0.033X

••00I/II
• I
ßr:0,960
(P<O,
01)
I I I I I • ' •
50 100
FIGURI;4.--Relationship between the chlorophyll
content of samplesof pond water and the chemical
oxygen demand of these samples.
o CHEMICAL OXYGEN DEMAND (Me/L)
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FIGURE3.---The relationshipbetweenchemicaloxy- sumption declined in sampleswhich contained

gen demandand the consumptionof dissolvedoxygen
by bacterial communitiesutilizing heat-killed plank- no living algae or zooplankton,oxygencon-
sumptionwas still at a significant level. Under
ton as a substrate.Sampleswere incubatedin the
dark for 24 hr at 30 C.
natural conditions, photosynthesisis nil im-
mediately following the sudden death of a
between15 and 35 C (Fig. 2). The increase bloom. Bacterial decompositionof the dead
was fairly linear for waters from ponds S-3, phytoplanktonand the combined respiration
S-6, and S-14 which contained moderate den- of other organisms soon reduce dissolved
sities of plankton. Pond S-11 contained a oxygen concentrationsbelow the critical level
densebloom of the blue-greenalga, Aphanizo- for fish survival. The rate of oxygenconsump-
menonfios-aquae.Respirationby this organ- tion reported in Figure 3 may be lower than
ism increasedrapidly between25 and 35 C. thatfoundat a similarCOD in a pondfollow-
The temperatureresponsecurvesyield Q10 ing a phytoplankton die-off. Bacterial pop-
values of 1.5 to 2.0. Oxygen consumption ulations in the heat killed samplesmay not
increasedby an averageof 1.74 times between have been present at as great a density as
20 and 30 C. Although depletion of dissolved found in natural waters. However, the con-
oxygen in pond waters occasionallyoccurs in sumptionof oxygenby bacteria growing on a
substrate of dead plankton increases as a
early spring, most instancesof oxygen deple-
tion are observed in the summer when water function of increasing COD.
temperatures are 28C or above (Swingle, A positive correlation existed between the
1968). amount of chlorophyll and concentrationsof
Much of the dissolved oxygen of water COD (Fig. 4). This indicatesthat even in
sampleswas consumedthrough plankton res- ponds which received applications of feed,
piration. Bacterial decomposition of heat- plant production within the pond was the
killed plankton within a 24-hr period con- major sourceof COD. Boyd (1972a) reported
sumedless oxygenper unit of COD than did mean rates of carbon fixation by photosyn-
the combined processesof plankton respira- thesisof 2.45, 2.49, 2.55 and 2.70 g carbon/
tion and bacterial decompositionin unaltered m2 per day in four ponds containing catfish
samples(Figs. 1 and 3). Observationson the which received heavy feed applications. As-
relationshipof phytoplanktondie-offs to suf- suminga value of 48% carbon in dried phy-
focation of fish suggestthat dead phytoplank- toplankton (Boyd and Lawrence, 1966) and
tondecomposes
rapidly(Swingle,1968). Data averaging values for the ponds, 5.32 g dry
in Figure 3 are not necessarilyin disagreement weight/m2 of organic matter was produced
with this hypothesis. Although oxygen con- daily. This is 53 kg/ha per day or 9,540 kg/
 610 TRANS. AMER. FISH. SOC., 1973, NO. 3

cl IOO MG OXYGEN REQUIRED TO COMPLETELY DXIDIZE

IMG DRY WEIGHT

PHYTOPLANKTON
CULTURES • (N= I0)

MACROSCOPIC-•I•-
ALGAE (N=4)

HIGHER
AQUATIC
PLANTS _• (N=9)
CHLOROPHYLL (JJG/L)
PARTICULATE J
MATTER (N=15)
FtcvaE5.--The relationshipbetweenthe chlorophyll
content of unfiltered pond water and the percentage
of the chemical oxygen demand which was found in FISH I (N=7)
flitrates of the samples.Sampleswere filtered through
glass-fiber filters. F[cua•; 6.--The amount of oxygenrequired to com-
pletely oxidize samplesof various aquatic organisms.
Horizontallines representmeans,vertical bars depict
two standard errors, and vertical lines indicate the
ha in a 180-day growing season. During the
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range. Samplesize (N) is givenaboveeach diagram.

same period, from 2,240 to 3,360 kg/ha of
fish feed is normally added to ponds contain-
ing catfish. The COD of high protein content apparently low in fat, had a COD of 0.97 mg
fish feeds (1.14 mg oxygen/rag dry wt) is oxygen/rag dry wt. Samplesof particulate
similarto that of phytoplankton(Fig. 6). The matter containeda mixture of phytoplankton,
efficiency of conversion of dry feed to dry zooplankton,and both organic and inorganic
fish is often as high as 25%, so only 75% of detritus. Wide variation in the COD of the
the COD of the fish feed would reach the water different samples of particulate matter was
as wastefeed or excretory products. However, causedby suspendedsoil colloids which were
nutrients from excretory products and waste retained on the filters. The low values 0.44
feed are responsiblefor the dense growths of and 0.53 mg oxygen/rag dry wt, were for
algae. waters which were fairly muddy. The lower
The COD of solubleorganicmatter (COD averagefor particulatematter as comparedto
in filtrates) in pond waters ranged from 6.2 that of algae from laboratory cultures was
to 43.9, averaging19.9 rag/liter. The percent- also related to the contamination of pond
age of the total COD in the soluble fraction waters with inorganic particles. For practical
ranged from 20.2 to 98.0%. The proportion purposes,a COD of 1.00 mg oxygen/rag dry
of the total COD which was not retained by wt may be assumed for aquatic plants and
filtration decreasedwith increasingconcentra- particulate matter from pond waters. Fish
tions of chlorophyll (Fig. 5). This further have a slightly higher averageCOD, but the
emphasizesthe importance of the contribution difference is not statisticallysignificant (two
of phytoplanktoncommunitiesto the COD. standard errors overlap).
Variation in COD of different speciesof COD values do not indicate the rate at
algae from unialgal cultures, macroscopic which organismswill decompose.Fish and
algae, and higher aquatic plants was fairly phytoplankton containmuch higher percent-
low (Fig. 6). Considerablymore variation ages of nitrogen than macroscopicalgae and
was encounteredbetweendifferent speciesof higher aquatic plants (Lawrence,1968) and
fish and samplesof particulatematter. Differ- will therefore decomposemore rapidly. These
ences in the COD of fish were apparently data indicate the total amount of oxygen that
related to the amount of fat in the samples. is required to decomposethe organisms. The
For example,the Ictaluruscatussample,which eventual oxygen requirements to decompose
containeda large amount of fat, had a COD
the standingcrop of planktonis large. In a
of 1.40 mg oxygen/rag dry wt. The Microp-
1-ha pond with an averagedepth of 1 m and
terus salmoidessample,which was dry and a planktonstandingcrop of 25 rag/liter, there
 BOYD--COD AND ORGANIC MATTER IN PONDS
is 250 kg of plankton. This will require an
equivalent amount of oxygen for complete
decomposition.
ACKNOWLEDGMENTS
This research was supported by Project
AID/csd 2780 to the International Center for
Aquaculture,Auburn University and Hatch
Project No. Alabama 287. The technical assis- HErliES,B. 1962. Primary productionin fish ponds
tance of Mrs. Lynda Tillery is appreciated. and its applicationto fertilization experiments.
Dr. H. S. Swingleoffered severalhelpful sug-
gestionsß
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wastewater.11th ed., New York. 626 p.
BASU,S.P. 1959. Active respirationof fish in rela-
tlon to ambient concentrationsof oxygen and
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BOYD,C.E. 1972a. Summer algal communitiesand
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ß 1972b. Sources of CO• for nuisance blooms
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, AND J. M. LAWaENCE.1966. The mineral
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R. G. 1936. Oxygenconsumption
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Blackwell Sci. Publ., Oxford. 172 p.
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LAWRENCE, J. M. 1968. Dynamicsof chemicaland
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river. Zool.-Ent. Dep. Series, Fisheries No. 6,
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PAMATMAT,M. M. 1968. Ecology and metabolism
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Int. Rev. Ges. Hydrobiol. 53: 211-298.
SAWYEa,C. N. 1960. Chemistry for sanitary en-
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SWINCLE,H. S. 1968. Fish kills caused by phyto-
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