Experimental Simulation of Winter Anaerobiosis in a Senescent Lake Reviewed work(s):

Experimental Simulation of Winter Anaerobiosis in a Senescent Lake
Author(s): Raymond L. Lindeman
Reviewed work(s):
Source: Ecology, Vol. 23, No. 1 (Jan., 1942), pp. 1-13
Published by: Ecological Society of America
Stable URL: http://www.jstor.org/stable/1930867 .
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No. 1
JANUARY, 1942
VOL. 23
EXPERIMENTAL
SIMULATION OF WINTER
IN A SENESCENT LAKE1
RAYMOND
L.
ANAEROBIOSIS
LINDEMAN
Department of Zoology, Universityof Minnesota
pursue furtherthe problem of winter
anaerobiosis. This lake originallyoccupied a basin of about 50 hectaresand was
at least 12 metersin depth. Encroachvegetation,followed
mentof mat-forming
by tamaracksand white cedars, has reducedthe lake to a merepond,Cedar Bog
Lake, 150 meterswide by 200 meterslong
and abouta meterin depth,locatedin the
middle of a dense bog forest. During
two or threemonthsof each winterthe
oxygenis exhaustedfromthe water beneaththe ice, presentingwhat seem to be
extremelyunfavorableconditionsfor the
entirebiota.
INTRODUCTION
The abilityof certainbenthicorganisms
to resistseasonalanaerobiosisin lake bottoms has long excited the curiosityand
admirationof biologists. A processanalogous to "holdingone's breath for several months" enables these forms to
thrivein the periodicallystagnatedooze
of eutrophiclakes,whichmightotherwise
supportonly micro-anaerobes.
Summerstagnationof eutrophiclakes
is believedto resultin completeabsence
of profundaloxygenfor as long as three
monthsor more. A shorterperiod of
benthicanaerobiosisoften occurs during
winterstagnation. In moreshallowlakes
HISTORICAL
and ponds withsoftooze bottomsthe latThe earlystudiesdealingwiththe fauna
ter stagnationperiod is far more important. In this type of lake thickice and of anaerobicprofundalareas were made
a heavy snow-covereffectively
shut out by Judayand his studentson the fauna of
the light necessary for photosynthesis; Lake Mendotaduringsummerstagnation.
bacterialdecomposition
and respiration
of Juday ('08) listed 11 species of protoaquatic organismsexhaust the dissolved zoons, 2 oligochaetes(Tubifex and Limoxygen; and veryoftenthe entireaquatic nodrilus), the nematodeAnguillula,the
theostracodCanChaetonotus,
habitatbecomes anaerobic. Such condi- gastrotrich
Pisidium
idahoensis,the
the
bivalve
dona,
tionsare oftenaccompaniedby spectacular
and
punctipennis
larva
Corethra
phantom
"fish-kills."
During a studyof seasonal population the midgelarva, Chironomustentans. In
dynamicsin a senescenteutrophiclake of 1919 he described an anaerobic ciliate
this type near Minneapolis (Lindeman, fromthissame zone,in whichhe reported
from13 to 17? C. and car'41), it was believed highlydesirableto temperatures
bon dioxidetensionof 4-9 cc./1.
1 This is the fourth in a series of papers,
Cole ('21) studiedthe respiratoryas"Ecological Studies of a Senescent Lake," depect of the problemon the midge larvae
scribing various ecological aspects of Cedar
and oligochaetesof the profundalhabitat
Creek Bog, Minnesota.
1
2
RAYMOND
L. LINDEMAN
of Lake Mendota. He suggested,but did
not prove,thatthesefacultativeanaerobes
get their respiratoryoxygen from the
atomic oxygen released by decomposing
plant tissues. This explanationhas been
consideredhighly improbableby recent
workers. Many otherstudies,bothbefore
and since,have been made on the physiologicalaspect of anaerobiosis.
in
Most of theorganismsoverwintering
Cedar Bog Lake must be consideredas
(von Brand and Harnisch,
euryoxybiotic
'33), possessingthe abilityto substitute
anaerobicprocessesforaerobicrespiration
for long periods of time. Harnisch
(1935-'39) has recentlymade exhaustive
studieson the physiologicalprocessesocand Tubifex durcurringin Chironormus
ing anaerobiosis. His work seems rather
to establishthatunderanaerobic
definitely
conditionsa certainamountof energymay
be released by the "splitting"of carbohydratesinto reduced substances. An
oxygendebt is thus builtup, whichmust
be oxidizedor "paid off"upon the return
of theorganismto aerobicconditions.He
has shown ('36) that Chironornus,released fromanaerobicconditions,supplewith
mentsits primaryaerobicrespiration
a secondaryoxybiosis. This secondary
process,conformingto the law of mass
action, proceeds rapidly to oxidize the
stored"Spaltprodukte."
Referencesto the occurrenceof microorganismsin habitatswithverylow oxygen contentare rathernumerous,although
definiteproof of anaerobicconditionsis
lackingin manycases. Lauterborn('08)
listedand discussedmanyspeciesof protozoons frompollutedareas of the upper
Rhine. Imel ('15) listed9 protozoons,a
and a Cyclopsfromthe hypogastrotrich
limnionof an Indiana lake. Rylov ('23)
of Loxstudiedthe ecologicaldistribution
odes rostrum,a ciliatewhichis abundant
in Cedar Bog Lake, and found that it
preferredwaterwithlow oxygentension,
but could not surviveanaerobicconditions
where HS was present. Noland ('25)
foundlow oxygentensionto be a limiting
of freshwater
factorin the distribution
Ecology,Vol. 23,No. 1
ciliates. Lackey ('25) listed 29 protozoons commonin sewageverylow in oxygen but high in carbon dioxide, and in
1932 listed 12 species "always presentin
the absence of oxygen in Imhof tanks."
Moore ('39) listed several groups as
knownto occur in the anaerobicsummer
hypolimnionof Douglas Lake: 7 protozoons, nematodes,2 rotifers,gastrotrichs,
3 copepodsand 4 ostracods. These organismswere reportedto "endureratherthan
select an anaerobicenvironment"
as Eggleton('31) concludedwithrespectto the
macrobenthos.
Experimentalstudieson resistancesof
organismsto anaerobiosishave been of
two types. The firstof theseconsistedof
laboratoryexperimentsin which certain
culturedorganismswere placed in synthetic"oxygen-free"chambers,and survival was timed under such conditions.
Bunge ('88) foundthat leeches,worms,
snails and arthropodsplaced into boiled
springwater in a stopperedvessel would
live from2 to 7 days under such conditions. The survivalof ciliates was first
studied by August Putter ('05), whose
methodof extractingoxygenconsistedof
boilingthe culturemediumand bubbling
pure nitrogenthroughit. All of theoxygen cannotbe removedby such a procedure,sinceoxygenis about twiceas soluble in water as is nitrogen,althoughthe
oxygentensioncan be reducedto a rather
small amount. His organismswere then
introducedintothe culturein a smallvolume (from which the dissolved oxygen
had not been extracted)equal to about 2
per cent of the volume of the culture.
No directchemicaltests were made for
dissolvedoxygen. Undertheseconditions
he found that Spirostomniu amnbiguum
could survivefor 24 hours,Paramecium
caudaturnfor 5-6 days, and Colpidium
colpoda for 16 days. From this he concluded that these organismswere conspicuouslyindependentof dissolvedoxygen for energy.
A veryextensivestudyof thistypewas
and Mudrezowaconductedby Nikitinsky
January, 1942
WINTER
ANAEROBIOSIS
IN
A SENESCENT
LAKE
3
Wyss ('30) in an attemptto correlate in the surroundingwater,and effectsof
the"saprotic"tendenciesof certainorgan- anaerobiosison reproduction. He conisms with their resistanceto CO2, H2S, cluded: "Considering the interplayof
and lack of oxygen. The methodsem- physical,chemical,and biologicalfactors
ployed sufferedfromthe same faults as of the environment,
it seems evidentthat
those of Putter, although to a much the pronounced effectivenessof the
greaterdegree,sincetheyused a streamof physical-chemical
stagnationis due not to
hydrogengas (whichis 30 timesless solu- the variationsof any one factorin the
ble in water than is oxygen,and which complex,but ratherto thecombinedaction
does not combinewith molecularoxygen of variationsof each factorin the presto formwater at ordinarytemperatures) enceof variationsof theothers. Evidence
bubbling through the culture water. now available indicatesthat the members
These workersmentionno chemicaltests of the profundalbenthicfauna are faculfor dissolvedoxygen,althoughtheyspe- tative ratherthan obligatory'anaerobes'
cificallystatethatoxygenwas "completely and thattheyendureratherthanselectan
absent." They concluded that "many anaerobicenvironment."
aquaticorganismsare able to endurecomplete absence of oxygen for 10-20-30
PREPARATION
OF CULTURES
daysand evenlonger. A fewciliateswere
The presentstudyis concernedwith a
able to multiplyunder such conditions.
simulation
of anaerobic lake bottomsin
The resistance against carbon dioxide,
winter.
The
methodsused are somewhat
hydrogensulphide and lack of oxygen
diminisheswith the complexityof the similarto thoseof Eggleton,althoughdeorganism:protozoansare more resistant veloped independently.The samples for
than rotifers,and rotifersare more re- theseculturesweretakenfromCedar Bog
sistantthan crustaceans." It seems quite Lake on Nov. 15, 1939. The lake at the
timeof collectionwas coveredwithabout
doubtful,however,thattheircultureswere
2 cm.of ice. Samplesof typicalsoftooze
trulyanaerobic.
bottomwere taken at a stationabout 25
A second typeof experimentation
was
metersfromshore. Eight Ekman dredge
initiatedby Juday. In 1908 he carefully
(225 cm.2 size) collections,continually
pumpedthe anaerobicooze fromthe bot- iced to
maintaina temperature
verynear
tomof Lake Mendotadirectlyintoculture 00
C., werebroughtbackto thelaboratory.
bottles,and foundthat the attendantorIn the laboratoryfour of the Ekman
ganismswould live in such culturesforat collectionswerecarefullysiftedthrougha
least two weeks at 13-15? C. Eggleton clean sieve (20 meshesto theinch) in the
('31) performedsome very informative same water fromwhichtheywere taken,
experimentson the macrobenthosof witha certainnecessaryadditionalamount
Douglas and Third SisterLakes in Michi- of the same bog water. The "filtrate"
gan. The typicalprofundalfauna was fromthesesiftingswas further
iced (with
composedof representatives
of thegenera ice cubes made fromsame water) and alChaoborus(Corethra), Chironomus,
Pro- lowed to standwhile the coarsermaterial
cladius (Protenthes),Limnodrilus,Pisid- containing
bottomanimalswas transferred
ium, Musculiumand Hydrornermis(?).
to a clean whitepan. The total macroBy placingknownnumbersof organisms benthicpopulationof the four collections
in culturejars of oxygen-freewater and was sortedout into iced fingerbowls,
and
replacingthemin the hypolimnion,
with foundto containa numberof organisms
manyvariationsof controls,he was able equivalentto the populationper square
to studythe toleranceand activityof dif- meterof lake bottomindicatedin the folferentspecies,chemicalchangesproduced lowinglist:
4
RAYMOND
Chaoborus punctipennis................
Chironomus plnimosus .................
Chironomus (decorus + lobiferus) .
Palpomyia sp. ........................
Limnesia sp .........................
Candona sp. .........................
Tubifex sp. .........................
3900
289
1676
222
33
56
222 2
The culture jars for the experiment
consisted of 12 glass-topped "Atlas" quart
jars which were thoroughly cleaned. The
outer surfaces of the jars and covers were
painted with two coats of black auto
enamel to eliminate any possibility of
photosynthesis.
The culture medium for the experiment
consisted of the selfsame ooze from which
the macrobenthos had been sifted, together
with all the attendant microorganisms
which had passed through the sieve. This
"filtrate" of semi-aerated ooze and bog
water was poured into a three gallon glass
bottle, thoroughly mixed, and, before settling could occur, was apportioned into
the 12 blackened jars, filling each to the
brim. The ratio of water to particulate
material was such that the settled ooze
occupied approximately one-half of the internal volume, giving it a depth of about
8 cm., almost the normal depth range of
The area of the
benthic ooze feeders.
ooze surface in each jar was found to be
64 cm.2
In an effort to establish in the culture
jars a population density approximating
that found in the lake, the following numbers of organisms were placed in each jar:
Chaoborus punctipennis ...................
Chironomusplumosus ...................
C. decorumand C. lobiferus ................
.........
Palpomyia sp. ..........
...........
Tubifex sp. ........
25
2
10
1
1
In addition, one Limnesia and one Candona were added to the samples set at
00 C. for 30 days and 60 days, and one
Limnesia was added to the 90 days culture set at 00 C.
The populated culture jars, sealed with
jar rubbers and the tops tightly clamped,
were then placed in a cool corridor (ca.
100 C.) for about 7 hours, after which 8
2
Others passed through sieve.
Ecology,Vol. 23,No. 1
L. LINDEMAN
of the jars were placed in carefullycontrolled temperaturechambers,4 jars at
50 C., and 4 at 100 C. The remaining
4
jars were placed in a pail and sunk into
thebottomof a shallowpond in whichthe
temperature
beneaththe ice was believed
to remainnear 0? C. The pond was icefree,withwaterat 50 C., on the relatively
warm day whenthe cultureswere placed
there; but it was coveredwithice a few
days later, and remainedso during the
restof the winter.
At the end of 30, 60, 90, and 120 days,
respectively,
one culturejar was removed
fromeach temperature
series for analysis
of its contents. Samplings were made
fromCedar Bog Lake at intervalsduring
the winterto checkagainstthese simulation cultures.
PHYSICAL
FACTORS
DURING
ANAEROBIOSIS
During the course of this experiment,
the temperatureof each series was kept
constantwithverylittlevariation(? 0.5?
C.) at 50 and 10?, whilethecultureseries
at 0? probablyremainedslightlyabove
that value. Color and turbidityof the
the samplewaterincreasedmarkedlyduring the anaerobic period; highestvalues
were foundin the sampleskeptat 100 C.,
indicatingmore dissolvedand suspended
substancesin thisculturethanin thoseat
the lower temperatures.
CHEMICAL
CHANGES
DURING
ANAEROBIOSIS
Only the mostsimplechemicalanalyses
weremade of theculturewater: pH, CO2
content,bicarbonatealkalinity,and dissolved oxygen. The pH content was
measured by comparisonwith LaMotte
bromthymolblue standards. Carbon dioxide contentand bicarbonatealkalinity
were measured according to procedures
suggestedin "Standard Methods for the
Examination of Water and Sewage."
Dissolved oxygen was measured by the
usual Winklertechnique. Hydrogensul-
January, 1942
WINTER
ANAEROBIOSIS
IN
A
SENESCENT
LAKE
5
phidecould not be detectedby odor when
Bicarbonatealkalinity
any of the jars were opened,even after
The benthicooze of Cedar Bog Lake
120 days of anaerobiosis,althoughall of
was
foundto containcarbonatesin excess
the culturesgave offa slightly"marshy"
of
1000
p.p.m.,indicatinga high alkaline
odor.
reserve. Thus as freecarbondioxidewas
Dissolved oxygen
produced,moreand moreof thecarbonate
was releasedto combinewithit as calcium
The water just above the ooze in the
bicarbonate. The increase in total alkashallowlake bottomwas about70 per cent
linityof thewateris therefore
a good supsaturatedwith oxygen at the time the
plementary
indexof theamountof carbon
original samples were collected. This
dioxide formedby decomposition. The
waterwas thoroughly
mixed withthe deendpointof this titrationwhen compared
composingooze and placed in the culture
with permanentstandards (brom cresol
jars. Because of theculturemethodused,
green) is a much more reliableone than
the exact time at whichoxygenvanished
the phenolphthalein
end point for carbon
fromtheblackenedculturebottleswas not
dioxide acidity.
preciselydetermined,
althoughsmall culIn this series of cultures,as shown in
tures withoutanimals were found to be
figure1, therewas a definiteincreasein
anaerobic after 24 hours of culturing.
alkalinity with prolonged anaerobiosis.
Ldnnerblad (1930), in studyingthe deAlkalinitychanges in the lake itself folcompositionof bottomooze in Swedish
lowed a similar course, although at a
lakes,foundthatooze of the "gyttja"type
much higherlevel, since anaerobiosisbeshowedonlya traceof oxygenat the end
gan muchlaterin the season. There was
of 12 hours when samples were placed
a greaterproductionof CO2 at 100 C.
under conditionssimilarto those of this
thanat either00 or 5?. A veryinterestabsent
study. The oxygenwas completely
factis thatthe data show greaterprofromthe experimental
jars whenthe first ing
of CO, at 00 than at 5?. This
series was removed30 days later,and it duction
the presence of certain
might
indicate
did not reappearin any of the cultures.
"cold-loving"bacteria,most active at 00
C., as suggestedby Henrici ('39). The
Carbondioxide
more unreliable carbon dioxide acidity
in thislight.
The difficulties
involvedin accuratede- data mightalso be interpreted
termination
of carbondioxide contentby
Hydrogen ion concentration
titrationare so familiarthat the matter
need not be elaboratedhere. Needless to
The hydrogenion contentof the culsay, in additionto errorsproducedby the
turesas expressedby pH varied from6.8
occurrenceof various unknown"humic" to
7.1 in an irregularmanner. Such minor
acids,etc.,the determination
of a "permachanges (possibly due to difficulties
in
nentpink" endpointwithphenolphthalein
matchingcolors with the murkywater)
in murkyyellowishwater was extremely
are believedto have no significance,
other
difficult.The authorattemptedto titrate
thanto indicatethatthe carbonatesystem
to a pink color which would remainfor
was adequatelybuffered.
one minute.
In spite of titrationdifficulties,
examSURVIVAL OF MACROBENTHOS
inationof figure1 will showthatthe CO,
acidityincreasedregularlywithlengthened Interestin this aspect of the problem
anaerobiosis,and that the amount pro- was the prime motive in beginningthis
ducedwas somewhatgreaterat thehigher seriesof experiments.Justhow long can
temperature.
benthicorganismstoleratethe absence of
6
RAYMOND
Ecology, Vol. 23, No. 1
L. LINDEMAN
74
7.0~~~~~~~~~~~~~~~~~~0
7?ta
LAKE
pH
15/0?
6.6
Tamne in dojS
30
90
60
120
50
/00
Oa?
C Oa
d o l
60
-40
200
0
0
Time in doys
30
90
60
120
360-LK
7ICO0
520
00
50
240
200160.
Pppm
0TMe; doajs
50
60
90
/20
FIG. 1. Alkalinity changes in anaerobic culture Jars.
oxygenin theirexternalenvironment
? It
seemed most logical to simulateas carefully as possible the species-ratiosand
densityfoundin thewildpopulation. For
this reason identicalpopulations,representingminiature"cross sections"of the
lake fauna,wereplacedin each culturejar.
When jars were removedfromthe tem-
peratureseries at the ends of their respectivecultureperiods,the supernatant
liquid was removedfor chemicalanalysis,
small amountsof ooze were diluted on
slides for microscopicexaminations,and
the remainingooze was siftedthrougha
finesieve (40 meshesper inch) foranalysis of macrobenthicsurvivals. The re-
January, 1942
WINTER
ANAEROBIOSIS
IN
A
SENESCENT
LAKE
7
mids encounteredby him in the Illinois
River region. In noneof the culturejars
examinedwas foundany evidenceof atDays
of
Temp. Chao- Chiron- Palpo- Tubi- Lim- Cantemptsof the larvaeto pupatein response
omus
fex
nesia dona
borus
myia
expo(C.)
to the oxygendeficit,
sure
as was suggestedby
Miall ('91) for Chironomussp.
1
1
1
0* 12
25
1t
The survivalsof Chironomuslarvae in
1
1
0
3
25
12
30
00
the lake were analyzed for comparison
2
50
11
1
25
30
with those of the simulative cultures.
2
12
1
25
30 100
Anaerobicconditionsprevailedbeneaththe
2
0
0
00
8
60
0
26t
ice fromFebruary7, 1940, to March 30,
0
50
1
60
5
19t
1940, a period of 51 days. A series of
1
25
4
0
60 100
samples
taken on January31 was con1
1
0
25
3
00
90
sidered to represent"initial" population
3
4
50
0
90
24t
1
0
2
90 100
2t
conditionsat the onset of anaerobiosisin
the
lake. Furthercollectionswere taken
1
0
00
11
2
120
3
1
4
50
11
120
on March 3 and March 30, twenty-four
0
1
0
1
120 100
and fifty-one
days,respectively,
afteroxygen had disappeared. No good criterion
* The initial population established in each
other than size could be found for the
culture jar.
t Initial population probably miscounted dur- separationof larvae of C. plumosusfrom
ing transfer.
C. decorum.It was found fromrearing
t Some small oligochaetes passed through the
sieve while being sifted.
experiments,however, that almost all
larvaegreaterthan 18 mm.in lengthwere
sultantsurvivalsof the macrofaunaare
C. plumosus,while adults reared from
shownin table I.
smaller
larvae were C. decorum.Larvae
The Chironomusfauna, consistingof
18 mm.,therefore,
longer
than
were condegreesof
mixedspeciesshowingdifferent
sidered to be C. plumosus,while those
toleranceto anaerobiosis,cannotproperly
be discussedas a genericunit,but rather
TABLE
II. Survivalof Chironomuslarvae in
species.
as a groupof ecologicallydifferent
anaerobiccultures
The relativetolerancesof the threemost
commonspecies are indicatedin table II.
Days of
Tempc C. plumosus C. decorousC. lobiferus
The initialpopulationsconsistedof 2 C.
0*
_
2
ca. 6
ca. 4
plumosus and 10 specimensof smaller
consistingmostlyof C. dechironomids,
00
30
2
4
3t
corus and C. lobiferusin an approximate
50
30
2
6
3
30
100
2
9
1
ratio of about 6 to 4. While the data
may be criticizedbecause of thisapproxi00
60
2
3
3
in50
60
mationand because of the statistically
2
2
1
100
60
2
2
0
sufficient
survivals,it seems clearlyindicatedthatC. plumosusis themosttolerant
90
2
1
0?
0
90
50
2
1
1
of extendedanaerobic conditions. Fur1
1
90
100
0
althoughthedata are notimpecthermore,
00
seemsto be moretolerant
cable,C. decorum
120
0
1
1
120
50
2
1
1
rethanC. lobiferus. These experimental
120
100
1
0
0
sults agree perfectlywiththe conclusions
of Richardson('28), who arrangedthese
* The initial population established in each
speciesin exactlythesame orderof "toler- culturejar.
t In addition, one specimen each of Limance to pollution"; these three species
nochironomus,Procladius and Tanytarsus was
chirono- recovered fromthis culture.
were the most pollution-tolerant
TABLE
I.
fauna in
Survival of macrobenthic
anaerobic cultures
8
RAYMOND
L. LINDEMAN
smallerthan this were consideredto be
C. decorum,although it was recognized
that a few immatureC. plurnosusmight
thus have been included. The paucityof
size (17-18 mm.),
larvae of intermediate
as indicatedin table III, is good evidence
were distinct.
thatthe species-populations
The populationdata shown in table III
Ecology,Vol. 23,No. 1
mortality
was partlydue to starvation,as
the foodorganismsof thisspecieshad disappeared from the 10? culture60 days
before; Frankenberg('15, p. 542), however, foundthat a Chaoboruslarva with
an irreparably
evertedpharynx(starved)
lived for about 50 days. The uniformity
of toleranceof the Chaoborusindividuals
appears in markedcontrastto thatof the
TABLE
III. Survival of Chironomuslarvae in
otherbenthicforms. The effectof 120
Cedar Bog Lake duringwinteranaerobiosis
days of culture was very evident; the
Numbers per 5 Birge-Ekman samples (1125 cm.2) populationseven at the lower temperatures seemed to be rapidlyreachingthe
"24
"IniLengt
5das
Length
limitsof their"oxygen debts." The intial"
days"
"513days
mm.gth
1-13-40 3-3-40
ferencemightlogicallyfollow that Cha5
12
3-4
0
oboruscannotwithstanda periodof win45
18
8
5-6
5
15
16
7-8
ter stagnation exceeding 4 months in
12
8
9-10
8
22
32
23
11-12
duration.
45
33
13
13-14
The Palpomyiasurvivals,based on but
12
21
15-16
6
3
2
4
17-18
a singlespecimenperculture,mustbe very
131
173
66 (38% survival)
Chironomus decorum
cautiouslyinterpreted.The appearanceof
7
19-20
12
4
2 specimensin the 90 days, 100 culture
4
21-22
7
8
6
11
4
23-24
was
probablydue to accidentalintroduc2
25-26
14
8
3
6
1
27-28
tion of an extra specimenwhen the cul20 (43% survival)
47
30
Chironomus plurnosus
tureswere established. The mere factof
survivalof all individualsplaced in the
most unfavorablecultures (120 days at
indicatethatC. decoruswas slightlymore 50 and
100), however, suggests anasusceptibleto 7 weeks of anaerobicfield erobic capacitieseven exceedingthose of
conditionsthanwas C. plumosus,although Chaoborus.
both sufferedhigh mortalities. Another
The initial Tubifex populationscould
of these data, of notbe preciselydetermined
possible interpretation
becauseof the
course,is thatsmallerindividualsare less passage of these slenderforms(or fragtolerantthanlargerones. Indicationsthat ments of them) throughthe meshes of
smallerindividualsof fishesare less tol- thecoarse sieve. A sufficient
numberwas
erantto low oxygentensionsthan larger retained,however,to assure the inclusion
ones have been foundin therecentstudies of one healthyindividualin each culture
of W. G. Moore ('40).
jar. It must be rememberedthat imwell adaptedto properorientationin the ooze or predaChaoborusis apparently
seasonal anaerobiosis. Apparentlyall in- tion by Palpomyiamay have been just as
dividuals (table I) were able to stand 90 effective
as anaerobiosisin causing those
days of cultureat the lowertemperatures. mortalitieswhichdid occur. The known
The variationsrecordedforthe60 and 90 survivals,of course,suggestthatTubifex
day survivalsmay well have been due to can withstand120 days of winteranaeroerrorsin countingthe initialpopulations biosis,a conclusionnot supportedby field
of thesetransparent
wrigglinglarvae dur- evidence,as we shallsee. Possiblywinter
ing transferto the culture jars. The predationwas more severe in the lake
at 10? for 90 days of than in these simulationcultures.
completemortality
Of the minorconstituents,
culture might have been the effectof
represented
highermetabolicactivitythan that pro- by such few individualsthat specimens
ducedat thelowertemperatures.Possibly could be includedonlyin the 0? cultures,
January, 1942
WINTER
ANAEROBIOSIS
one Limnesia was able to endureanaerobiosis for90 days. Its toleranceof anaerobic field conditions (see below) supports this evidence of high resistance.
Candona, on the otherhand, was unable
to surviveeven 30 days in the cultures,
althoughdeath mighthave been due to
rough treatmentduring transferto the
culture jar. Field data, however, support the conclusionthat Candona is not
verytolerantof anaerobicconditions.
IN
A
SENESCENT
LAKE
9
robicconditionsthattubificid
oligochaetes
and that tubificidscould withstandexperimentalanaerobiosismuchlongerthan
Chironomnus,
althoughH2S was present
underhis experimental
conditions. These
statements
do notseemto agreewithwhat
has been found in the Cedar Bog Lake
and culturessimulatingit. The speciesof
Chironomusdifferedin their resistances
nearly as much as did the three major
groups studied by Alsterberg. In the
Cedar Bog Lake cultures Chaoborus
fauna in
IV. Survival of macrobenthic
TABLE
seemedto be more resistantthan ChiroCedar Bog Lake duringwinteranaerobiosis
nonus
decorusand Chironomuslobiferus,
Numbers per Birge-Ekman sample (225 cm.2)
but it appeared definitely
less tolerantof
120 days of anaerobiosisthanChironomus
"24 days"
"51 days"
"Initial"
Genus
plumosus,Tubifex and Palpomyia.
2.8
17.24 3.4
38.54
44.04 9.0
Chironomus.
0.0
2.5
2.54
2.2
Candona ...
0.8
The remarkabletoleranceof these or0.5
1.04 0.7
6.0 A: 1.8
5.2:i
Tubifex........
ganisms is eloquent evidence of their
0.2
1.64 0.7
0.24
Procladius ....
1.44 0.5
adaptationto a seeminglyintolerableen6.2:4
1.3
3.14 1.0
4.8:4 0.7
Palpomyia .....
1103.041117.0 835.6 473.5
Chaoborus..... . 715.8 477.1
vironment.The survivalof speciesunder
experimental
conditionsfar moreinimical
Partial correlations of the general than would normallybe found in nature
benthicsurvivalin the cultureswith the makes a complete"winter-kill"of these
survival of macrobenthicforms in the species quite unlikely.
lake were made possible by a series of
winterfield populationsamplings. The
SURVIVALOF MICROBENTHOS
figuresin table IV representthe numbers
When the simulationcultureswere esof individualsper 225 cm.2,expressedas
tablished,
much superfluoussifted ooze
means ? standard errors of the means,
remained.
This ooze was placed in a cool
based on five separate Ekman dredge
at
120 C. fortwo days,folcorridor
about
samples
These
date.
samples for each
were taken as follows: on Jan. 31, 1940, lowing which a study was made of the
seven days beforeanaerobiosisbegan and contained organisms. The microfauna
abundant
consideredas the "initial"anaerobicpopu- was foundto be so surprisingly
lation; on March 3, 1940,after24 days of that the ooze surface consisted,almost
anaerobiosis; and on March 30, 1940, literally,of "ciliate soup." The author
after51 days of anaerobiosis. These data is indebtedto thelate Dr. JohnP. Turner
of species. The
correlate quite well with those of the for aid in determination
with marks of
following
list,
together
in
the
culanaerobiosis
of
first60 days
tures. Chironomus,Candona, and pos- relativeabundance,representsthe species
to observed:
siblyTubifexseemedto be succumbing
the unfavorableconditions. The other
Acanthocystissp.
forms(includingProcladius culiciformnis,
Arcella vulgaris
not presentin the cultures) seemedto be
Centropyxissp.
"holdingtheirown." As anaerobicconDinamoeba sp.
ditionsin the lake were relievedat this
Euglena deces +
point, no furthercorrelationscould be
Glenodiniumcinctunm
made.
Heteronemaacus ++
Alsterberg('22) reportedthat ChaoHeteronema sp.
borus was much betteradapted to anae-
10
RAYMOND
L. LINDEMAN
Ecology,Vol. 23,No. 1
this group of culturesbut could not be
foundin later cultures.
The culturesopened at the end of 90
Bursaria truncatella
and of 120 days of anaerobiosiscontained
Coleps hirtus+
an even furtherdiminutionof microFrontonialeucas +++
organisms. Several 1 cc. sampleshad to
Loxocephalus granulosus ++
be studied in order to find even a few
Loxodes rostrum+++
individuals. Nematodes, copepods and
Parameciumcaudatum+
smallostracodsstillsurvivedin the cooler
Spirostomuntambiguunm
+
90 day cultures,but could not be found
Spirostornumteres +
after120 days of anaerobiosis.
Stentorcoeruleus+
Field correlations,made by comparing
culturedpopulationswithcorrespondthe
Microscopicmetazoonsobservedwere
ing naturalones in thelake,indicatedthat
Chaetonotusmaximus
at the beginningof anaerobiosisin the
Chaetonotusspp.
lake the populationwas essentiallyidenLepidodermiarhomboides
ticalwiththatoriginallyestablishedin the
Bdelloidea (Rotifera)
culturejars. At the end of 51 days of
Notommatoidea(Rotifera)
anaerobiosisthefieldpopulationshoweda
Nematoda
markedquantitativediminution;roughly
Cyclops sp.
estimated,the lake fauna showed only
Cypria sp.
about 1/20 of its pre-anaerobicdensity.
The species ratiosdid not appear to have
Additionalspecies later found in the anchanged as a result of these anaerobic
aerobicculturejars were as follows:
conditions,except that the cryptomonad
flagellatesseemed relativelymore abunpalustris
Pelomyxa
dant and variedthanbefore. No marked
Metopus spiralis
qualitative changes were apparent. As
Holotricha(tiny ciliates)
spring thaws relieved the oxygen defiHypotricha(tiny ciliates)
very shortlyfollowingthis obserciency
Cryptomonada(flagellates)
vation, it was impossibleto study the
At the end of 30 days of anaerobiosis effectsof more extendedanaerobiosison
fauna keptat 00 and at the naturalmicrobenthos.
the microbenthic
50 remainedvirtuallyunchangedover its
initialconcentration.The culturekeptat RE-AERATEDCULTURESOF MICROBENTHOS
onlya few
different;
100 was remarkably
For the purposeof studyingthe extent
individualsof the hardiergenera, Fronformscould encyst
whichmicrobenthic
to
were
tonia, Coleps and Heteronema,
afterthe onset
stages
resistant
or
produce
found.
two series were esAt the end of 60 days, the abundant of oxygendeficiency,
ciliate fauna of all three series was re- tablishedfromooze whichhad been under
ducedto a mereremnantof hardierforms. anaerobic conditionsfor 90 days. One
amoeba,Pelomyxa palus- series had been kept at 0? and the other
The interesting
in two of these cultures. at 100. Four small sample bottlesconfound
tris,was
The sluggishnessof this species and its taining ooze and oxygen-freeculture
general resemblanceto fecal balls of water in the ratio 1: 4 were established
chironomidsmade it rather difficultto for each series. These bottleshad been
distinguish. It may have been present sterilizedpreviouslyand the ooze was
in some of the othercultures,but could added in such a mannerthat there was
not be found in spite of specificsearch no undue exposure to air-borne cysts.
were easily foundin Covers were set looselyon the bottlesso
for it. Gastrotrichs
Phacus pyrum+
Trachelomonaseuchora
January, 1942
WINTER
ANAEROBIOSIS
IN
A
SENESCENT
LAKE
11
that air could enter; the samples were tendedanaerobiosisare not able to withthen placed in a constant temperature stand such conditionsby means of cysts,
room at about 150 C. That the samples but that when a few are able to survive
had actuallybecomere-aeratedwas shown such extremeconditionsand are subseby an oxygentest made after33 days of quentlyplacedin a favorableenvironment,
these cultureconditions:the water con- theywill multiplyquite rapidly. Also, as
tained 4-5 p.p.m. of dissolvedoxygen.
Lackey ('32) suggests,thereseem to be
The culturesfrom the 00 series con- certaincolorlessflagellateswhichare able
tainedno trace of microorganisms
except to grow in nearlyanaerobiccultures,and
for a smallcolorlessflagellateat the time which are unable to withstandre-aeratheywere transferredfromanaerobicto tion. Since such flagellatescould not
aerobicconditions. At theend of 13 days have gained entranceto the culturejars
this series contained almost as great a after anaerobiosis had set in, possibly
paucityof organisms. Study of a single theycame fromcystsformedunderaerosample fromeach of four duplicatebot- bic conditionsand re-encystedwhen the
tles revealeda totalof only2 smallnema- cultureswere again suppliedwithoxygen.
todes, 2 individuals of Heteronema, 1 The numbersencountered,
however,were
Coleps and 2 rotifers,Hydatina and too small to verify such a conclusion.
Synchaeta. The protozoonsmightpos- The scanty evidence available suggests
sibly have been overlookedin the initial that possiblyrotiferanand gastrotrichan
survey. A similar examinationat the eggs are able to surviveanaerobiosisand
end of 33 days revealed a considerable to emergeas adultsundermore favorable
increase in the numbersof Coleps and conditions.
Heteronema. A few hypotrichciliates
were observed;thesemighthave come in
CONCLUSIONS
as contaminationfrom the air. Also 2
1. Anaerobicbenthoswas studied exspecimensof thegastrotrich,
Chaetonotus,
were observed. The minute flagellates perimentallyunder winter conditionsat
presentin the anaerobicculturecould no 0?, 5? and 10? C. for 120 days, with
respectto survivalof macro-and microlongerbe found.
The culture from the anaerobic 100 organismsand variationsin theirchemical
seriesshowedan almostcompleteabsence environment.This was correlatedwith
fieldconditionsin a senesof organisms. A search lasting more corresponding
cent
lake.
than an hour revealed only 3 minute
2. Carbon dioxide content increased
flagellatesand a nematode. At the end
of 13 days under aerobic conditionsthis with extended anaerobiosis,and became
series seemedto contain,as far as could higherat 100 thanat lowertemperatures.
be observed,onlyrotifers:a bdelloidspe- More carbondioxide was producedat 0?
cies and an extremelyshort-spined"hae- than at 50; this suggeststhe presenceof
mal" variety of the form describedby "cold-loving"bacteria.
3. Organisms were more resistantto
the author('39) as Brachionushavanaenanaerobiosisat 00 and 50 than
extended
sis var. minnesotensis.At the end of 33
at
because of slower
100,
presumably
days this series showed nothingbut a
very few small hypotrichs. The minute metabolism.
4. Specimens of Chaoborus, Chiroflagellates had apparently disappeared.
and Tubifex were able
Thus, in spite of abundantoxygen and nomus,Palpomnyia
presumablyfavorable temperaturethere to survive 120 days of anaerobiosisin
was no reappearanceof the large initial mixedpopulations. The threemostcomholotrichpopulationfound in the lake.
mon species of Chironomusshowed the
These re-aeratedculturesthus seemed followingorder of tolerance: plumosus
to show thatprotozoanssubjectedto ex- > decorus> lobiferus.
12
RAYMOND
L. LINDEMAN
Ecology,Vol. 23,No. 1
1936. Primdre und sekunddre Oxy5. Microbenthicorganisms gradually
biose der Larve von Chironomus thummi.
disappearedwith extended anaerobiosis;
Ibid. 23: 391-419.
certainsmallcolorlessflagellatesappeared
1937. Primare und sekundire Oxyto be most resistant.
wirbelloser Tiere.
der Larve
biose
and Mudre6. The theoryof Nikitinsky
Verh. deutsch. Zool. Ges. 39: 129-136.
1938. Studien zum Anaeroben und
zowa-Wyssthatresistanceto anaerobiosis
der Larve von
Erholungsstoffwechsel
did
complexity
diminisheswith structural
Chironomus thummi. I. Wechsel im Glynot hold for these cultures. Certain
Zeitschr.
cogen-, Fett- und N-Gehalt.
nematodes,copepods and ostracods far
vergl. Physiol. 26: 200-229.
outlastedmost of the protozoons.
1939. Studien zum Anaeroben und
von
Erholungstoffwechsel der Larve
7. Preliminaryexperimentswith reChironomusthummi. II. Saurebildung und
aeration cultures seemed to show that
Ibid. 27: 275-303.
"Notoxybiose."
protozoansare not able to withstandan1939. The distribution of
Henrici, A. T.
aerobic conditions by means of cysts,
bacteria in lakes. "Problems of Lake Biolwhile eggs of rotifersand gastrotrichs ogy." A. A. A. S. Symposium No. 10:
are able to surviveand later hatchunder
39-64.
Imel, H. G. 1915. Some preliminary obsera more favorableoxygensupply.
vations on the oxygenless region of
8. The resultsseemed to indicatethat
Center Lake, Kosciusko County, Indiana.
none of the organismsstudiedwould be
Proc. Indiana Acad. Sci. 1915: 345-356.
able to endure anaerobiosis indefinitely,Juday, C. 1908. Some aquatic invertebrates
with the possible exception of certain
that live under anaerobic conditions.
Trans. Wisconsin Acad. Sci., Arts, Lett.
colorlessflagellates.
16: 10-16.
. 1919. A fresh water anaerobic ciliate.
Biol. Bull. 36: 92-95.
Alsterberg, G. 1922. Die respiratorischen Lackey, J. B. 1925. The fauna of Imhof
Mechanismen der Tubificen. Eine experitanks. New Jersey State Agric. Exper.
mentelle-physiologische Untersuchung auf
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1932. Oxygen deficiency and sewage
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Standard methods for the examination of
Rhizopoden und Infusorien aus dem
water and sewage. 8th ed. New York.
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Brand, Th. v., and 0. Harnisch. 1933. Die
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Einteilung der Tiere nach der Eigenart
Lindeman, R. L. 1939. Some affinitiesand
ihres Betriebsstoffwechsels. Zool. Anz.
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1941. Food-cycle dynamics in a
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Cole, A. E. 1921. Oxygen supply of certain
636-673.
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G. 1930. tVber die SauerstoffLonnerblad,
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Moore, G. M. 1939. A limnological investiFrankenberg, G. 1915. Die Schwimmblasen
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Harnisch, 0. 1935. Versuch einer Analyse
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CITED
-
January, 1942
WINTER
ANAEROBIOSIS
tions. Unpublished Ph.D. Thesis, University of Minnesota.
Nikitinsky, J., and Fr. K. Mudrezowa-Wyss.
1930. tVber die Wirkung des Kohlensaiure, des
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A
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LAKE
13
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im Wasser geldsten Sauerstoffs und
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