Hydrobiologia, vol. 39, 3, pag . 405-427, 1972

A Test of the Cosmopolitan Distribution ofFresh-Water Protozoans

by

JOHN CAIRNS JR. & JEANNE A. RUTHVEN,

Biology Department and Center for Environmental StudiesVirginia Polytechnic Institute and State University

Blacksburg, Virginia 24061 U .S .A .

INTRODUCTION

The cosmopolitan distribution of fresh-water protozoans is openlyacknowledged by many protozoologists and other microbiologists .Others tacitly acknowledge cosmopolitan distribution when theyuse such keys as those of KAHL (1935) or PASCHER (1913, 1927) inthe United States or use keys developed in the United States, inEurope, Africa, or other parts of the world . If protozoans did nothave a cosmopolitan distribution, then regional keys would beessential. Early recognition of a very broad distribution of someprotozoan species was provided by DARWIN (1860) in The Voyageof the Beagle. Aerial transport of microbial species has been studiedmore recently by such investigators as BROWN et al . (1964), MAGUIRE(1963), and SCHLICHTING (1961, 1964) and through animal move-ments by MAGUIRE & BELK (1967) . Some of the benefits of a broaddistributional pattern have been discussed by CAIRNS (1967) . Morerecently, MAGUIRE (in press) has studied the invasion of sterilewater in containers on Surtsey .

Despite the potential significance of the possible cosmopolitandistribution of protozoans and the ecological consequences of sucha distributional pattern, remarkably little research has been carriedout to test this possibility . It occurred to us that one suitableenvironment to test the hypothesis of cosmopolitan distribution offresh-water protozoans would be an island containing very littlesurface fresh water and surrounded entirely by salt water . In

Received March 13, 1971 .

405

addition, it would be helpful if this island were a considerabledistance from a large continental land mass . Accordingly, when anopportunity arose to visit Abaco Island in The Bahamas Chain, wedecided that this was a very suitable situation to test the hypothesisof cosmopolitan distribution . Although it would have been desirableto have a smaller and more isolated island, containing less freshwater than Abaco, the situation was nearly ideal . We felt that ifsterile containers with sterile water placed in various positionsaround the island were colonized by fresh-water protozoans, and ifthe soil of the island which only intermittently received fresh waterrainfall provided potential colonizing species, this would be evidencethat colonization could occur in areas not normally hospitable tofresh-water protozoan species .

Although Abaco Island is rather large, these investigations werecarried out at one tip of the island, Sandy Point, which usuallyreceives breezes from the ocean . In addition, the nearest pond wasquite a few miles away.

METHODS AND PROCEDURES

This investigation was divided into three operational categories(1) Samples taken from small bodies of surface water, exposed toair currents, etc . ; (2) A series of 24 sterile jars containing sterilewater placed at various locations (Fig. 1) on Sandy Point, AbacoIsland, Bahamas ; and (3) An attempt to obtain cultures of fresh-water protozoans from surface soil samples collected at variousspots on the Sandy Point area of Abaco Island (Fig . 1, 2, and 3) .Since the methods and procedures for each of these are quitedistinct, they will be handled separately .

Examination of Surface WatersExamination of various surface waters in the vincinity of Sandy

Point, Abaco Island, was made during the latter part of December,1969, in order to obtain some estimate of the kinds of species offresh-water protozoans found in these waters . Samples were col-lected by sucking material from the substrate with a clean rubberbulb and placing the sample material into clean, half-pint, screw-top jars. These samples were immediately taken back to the fieldlaboratory for examination. A few conventional stains and taxo-nomic keys were used for identification. Due to the limited numberof keys that it was possible to transport on the visit, some identifica-tions could be made to genus only but wherever possible, identifica-tion to species was carried out . Algal collections were made simul-

406

Fig. 1 . Map of southern tip of Great Abaco Island with enlargements of SandyPoint and the Duck Pond . The points 1-24 at Sandy Point show locations of thejars used in the protozoan colonization studies . The lacation of the 4 Duck Pondsamples and 4 soil samples (23-26) are also included .

taneously with the protozoan collections by DR. CHARLES W.REIMER of the Limnology Department of the Academy of NaturalSciences. Portions of these algal samples were also made availablefor evaluation of the protozoans they contained . At the same timethat the biological sampling was carried out, water samples weretaken from each location for chemical and physical determination .Some of these, such as dissolved oxygen concentration, pH, andtemperature were carried out on the spot . Others, such as totalhardness and chloride concentration, were carried out later in the

407

CEMETERY RD .

BAY ST. W .mm.J NOT A

WELL ; OPEN

PIT IENINDM

CHURCHN .RSOUR L .

aA2

DAY !T E

E WINDMILL WELL

408

SANDY POINT

TIP OF SANDY

POINT

Fig. 2 . Map of Sandy Point showing location of wells. Tip of Sandy Point withlocation of five soil sample sites .

field laboratory. It should be emphasized that the main purpose ofthese collections was to determine the variability of aggregations ofspecies of communities in various surface fresh waters of the SandyPoint region, rather than to make an exhaustive inventory of thespecies found in that part of the island .

2. Colonization StudiesTwenty-four sterile jars containing boiled, cooled water from a

well at the tip of Sandy Point were used to make a preliminaryestimate of the colonization potential of fresh-water protozoans inthis area of the world . The containers used were one quart masonjars with screw-top lids . The lids were discarded and the jars aloneplaced in boiling water for 20 minutes, and then allowed to cool .Water from the Sandy Point fishing camp well (for water quality

SANDY POINT - CAMP

LOCATION OF SOIL SAMPLES 1-21

TFig . 3 . Map of section of Sandy Point showing the location of soil samples 1-21 .

WM

w

El-F±] = CABINS

•

= BOAT HOUSE

•

= DOCK

C = CLUB HOUSE WELL

DW = DRINKING WELLGS = GENERATOR SHED

OH = OLD HOUSE

OW = OLD WELL

•

= STORAGE

WM-W= WIND MILL WELL

409

see Table IV - location B) was also boiled for twenty minutes,allowed to cool, and then poured into the jars in such a way thatmaximum aeration would be achieved . These jars without lids wereplaced in various spots throughout the fishing camp and the villageof Sandy Point . An attempt was made to place the jars so that theywould not get direct sunlight for the entire day and become over-heated. Most of them were placed under vegetation or in some otherspot at least partially protected from direct sunlight . The initialplacement was made early in December, 1969, and the contentswere examined three weeks later. Each jar was brought back to thelaboratory and allowed to sit for approximately one hour near thewindow to permit the protozoans to aggregate at the top andbottom. At that time, examination of the jar began with samplesbeing taken from the meniscus toward and away from the light, andon the bottom toward and away from the light . Two examinationswere made from each of these areas for each jar .

3. Soil SamplesSoil samples were taken at various locations on Abaco Island,

Bahamas with most of the samples (1-21) coming from SandyPoint . Samples 1-21 consisted of a gray, sandy soil with someorganic material, 23 & 24 were clay soils, 25 was a red-brown finesand, and 26 was mostly dark humus. Four mud samples were alsotaken from a brackish pond. Soil samples were collected in sterileNasco Whirl-Pak plastic bags and sealed to prevent outside con-tamination. Cultures from samples 1-21 were started about 1week after collection. The other samples were collected later andreceived by the authors about one month after collection . Culturesfrom these collections were begun several days after receiving them .

Sterile 25 ml test tubes were used for culturing . The followingtypes of cultures were established for each soil sample :

Distilled water to1000 ml

1 boiled wheat grain

In order to avoid excessive contamination in setting up the cul-tures, soil was transferred directly from the collection bag to thetest tube and 3-4 ml was estimated, not measured. One bag(Duck Pond 1) was torn when received, but this sample was still

41 0

I . II. III .

3-4 ml soil 3-4 ml soil 3-4 ml soil15 ml distilled water 15 ml Chu-14, modified 15 ml inorganic solution0.05 g powered milk (Chu, 1942) NaCl

0.1 gKCl

0.004 gCaCO, 0.006 g

used. Soil was first added to the test tubes, and then the culturefluid. The two were mixed by shaking the tubes, after which theywere stoppered with cotton plugs . Cultures were maintained atroom temperature (23-24°C) and about 8-9 h of light/day whichcame from overhead lights in the lab. The samples were left un-disturbed for a week and then examined once a week for 3 weeks ;however, during the second week not all of the tubes were examined .Before examination each tube was shaken slightly and a smallamount of water withdrawn near the surface of the soil substrate .One slide (about 2 drops) was made from each culture .

Each organism was identified to species whenever possible ;otherwise it was identified to genus with a drawing or descriptionmade to aid in differentiating species . No stains or fixatives wereused in identifications, but methyl cellulose was sometimes used toslow down rapidly swimming protozoans. No attempt was madeto estimate density although in a few cases the density of a singlespecies was quite high (5 10 individuals per 10 x field of view) asfor Mastigamoeba invertens, DPIW, second week ; and Tetramitis pyri-formis, DP2W, third week .

RESULTS

1 . Examination of surface waters - results are given in Table Iand the water quality and elevation data for some of these locationsin Tables II-IV . (See Fig . 2 for well locations) .

2. Colonization studies - results are given in Table V and thewater quality and elevation data in Tables VI, VII and VIII .

3. Soil samples - results are given in Tables IX and X . DP indi-cates samples from the Duck Pond, Abaco Island. M indicates theorganism was found in the milk culture ; C, in the Chu-14 culture ;and W, in the boiled wheat culture . There was no sample number22. Data from the 3 weeks were combined in Table IX . Altogether39 species of Mastigophora, 15 species of Sardodina, and 67 species ofCiliata were found. In cases where there is a question about speciesidentification, there is either a question mark or an indication thatthe organism appears to be similar to a certain species . Table Xgives the number of species found at each examination .

DISCUSSION

1 . Examination of surface waters . The evidence provided sug-gests that even in areas remote from significant surface fresh water

41 1

412

Table I

Collections from Representative Fresh-Water Habitats

on Southern Abaco Island

Subclass ZoomastigiaOrder RhizomastigidaFamily Mastigamoebidae

Mastigamoeba up .

XOrder ProtomonadidaFamily Amphimonadidae

Dinomonas vorax Kent

XFamily Bodonidae

Bodo emoeblnns Lemme

x

X,B-._ caudatus Dr0j .

XB,,, obosus Stein

X$y lens (Miller)

XH, obovatus Lemm .

XL- BP .

X X XOrder PolymastigidaFamily Hexamitidae

))examita up .

XOrder TrichomonadidaFamily Trichomonadidae

Tritriohomonas au¢ueta (Alexeieff)

X.L_ batrachoru% (Party)

X

1 5

A* B C D E F 2 3 4 G M I J 6 7

8

Phylum ProtozoaSubphylum PlasmodromaClass MastigophoraSubclass PhytomaetigiaOrder ChrysomonadidaFamily Chromulinidae

Oikomonas term (Eh-) XPseudochromuline sp . xChrysopsia sp . XPyramidochrysis sp . X

Family OchromonadidaeMoons sp . X X X

Order CryptomonadidaFamily Cryptomonadidae

Cryptomonas erosa Ehr . XC . ovate Ehr . X X XC- sp . XChilomonas paramecium Ehr . X

Order PrytomonadidaFamily Chlamydomonadidae

Chlamydomonas globosa Snow XC . sp . X X

Family CarteriidaeCarteria sp . X

Family VolvocidaePandorina morum Muller X

Order EuglenoididaFamily Euglenidae

Euglena acus Ehr . X X XE . fusca (Klebs) XPhacus acuminata Stokes XP . elate Klebs XP . pleuronectes MullerEutreptia viridis Party XE . sp . X x

Family AstasiidaeRhabdomonas sp . X

Family AnisonemidaeAnisonema emarginatum Stokes XPeranema granullfera Penard XP . trichophorum (Ehr .) XHeteronema sp . XNotosolenus sinuatus Stokes X

Order DinoflagellidaFamily Gymnodiniidae

Hemidinium nasutum Stein XAmphidinium sp . X

Family PeridiniidaePeridinium,tabulatum Clap .&

Lach . X

J- sp . XGonyaul x apiculata7 (Penard) X

Pleuronema sp .Cyclidium sp .

Subclass SpirotrichaOrder HeterotrichidaFamily Plagiotomidae

Nyctotherus cordiformis (Ehr .)Order Oligotrichida

Halteria ,grandinella (Muller) XOrder HypotrichidaFamily Oxytrichidas

Oxytricha bifaria Stokes

X0 . sp .

XAmphisiella sp .

Family AspidiscidaeAspidisca costata (Du) .)

XA . lynceus Ehr .

X

X

Subclass PeritrichaOrder PeritrichidaFamily Vorticellidae

Vorticella campanula Ehr .

X X

XV . convallaria Ehr .

X

V . Faure-Fermiet

XOrder OpalinidaFamily Opalinidae

Opalina sp .

X

*1. Abandoned main well near tip of Sandy Point, sampled 12/26/69 .

A. Associated with bottom mud .E . Associated with slime on submerged board .

(to be continued on pag . 427)

4 1 3

A D C D E F 2 3 4 G H I J 6 7 8Class SarcodinaSubclass RhizopodaOrder AmosbidaFamily Naegleriidae

Naepleria type xFamily Amoebidae

Amoeba eroteus (Pallas)Vahlkamofia lima. type x X XHartmanella astromvxis ?

Ray and Hay .. Xseveral small amoeba x

Order TestacidaFamily Difflugiidae

Difflueia oblongs Ehr .Subclass ActinopodaOrder HeliozoidaFamily Actinophryidae

Actinophrys sol Ehr . X

Subphylum CiliophoraClass CiliataSubclass HolostichaOrder GymnostomatidaFamily Holophryidae

Lagynophora sp .Enchelys curvilata (Smith)

Family ColepidaeColepS hirtus (Muller) X

Family AmphileptidaeLionotus cygnus (Muller) XL . trichoc stis Stokes X

Family ChlamydodontidaeChilodonella cucullulusC . fluviatilis Stokes

Family NassulidaeNassula sp . X

Order TrichostomatidaFamily Colpodidae

Colpoda sp . X X XTillina sp . X

Order HymenostomatidaFamily Tetrahymenidae

Colpidium campylum (Stokes) XFamily Frontoniidae

Frontonia leucas Ehr . X

Cinetochilum margaritace_Petty x X X X X

Family Pleuronematidae

*Well Sites (See also locations on map - Figure 2)

414

Table II

Water Chemistry Data for Surface Waters, Sandy Point, Abaco Island

Sample No . 1 2 3

4 6 7 8

J

pHD .O .AlkalinitySpecific Cond .Hardness (tot)PhosphateN0 3 ppmS04 ppmCl ppmCaMgFeCuZnCr

(See Table

Sample Location

ppm

I footnote

B

8 .36 ppm240485214 .00 .750 .653 .3516 .3676215000 .040

C

for explanation

8 .35 ppm295624213 .60 .160 .603 .7545 .91121920000

D

8 .153 ppm

130

2721438

680262 .6

221.40 .32

0.1031 .75

1.3315 .3

2.25332

79.6130

133288

1000

00

00

0.060

0

of sampling sites)

Table III

Well Water Quality*

8 .59 ppm100328101 .60 .1150 .651 .5051 .03844000 .040

G

8 .12 ppm1702650344 .60 .120 .9820 .5668 .01384150000

H

7 .33 ppm2132010337 .60 .150 .6515 .0516 .0182340000 .040

I

pH 7 .63 7 .65 7 .72 7 .78

7.45 7 .55 7 .50 7 .70 7 .88Alkalinity 295 255 270 235

265 195 270 175 155Specific Cond . 540 600 555 445

456 410 645 397 318Hardness (tot) 259 .0 239 .0 243 .9 239 .6

225.1 168 .8 186 .8 176 .4 127 .5Phosphate ppm 0 .10 0 .08 0 .08 0 .075

0.075 0 .20 0 .13 0 .08 0 .13N03

ppm 0 .15 0 .65 1 .65 1 .30

1.28 11 .0 1 .35 14 .9 7 .1S0 4

ppm 7 .5 7 .56 4 .5 4 .4

3.9 4 .5 6 .0 6 .75 6 .0Cl

ppm 8 .12 46 .0 33 .7 13 .3

9.73 12 .80 55 .10 10 .26 14 .32Mg 237 171 201 195

156 ill 165 109 109Fe 0 0 0 0

0 0 0 0 0Cu 0 0 0 0

0 0 0 0 0Zn 0 0 0 0

0 0 0 0 0Cr 0 0 0 0

0 0 0 0 0

B Well next to club houseC Drinking Water WellD Well in back yardE Windmill wellF Well behind old house next to windmillG Napoleon's wellH WellI JackJ Open

at old house behind schoolHarding's wellpit behind Anglican Church

Table IV

Water Quality Data - Duck Pond - Abaco Island

Bahamas DP 1

DP 2

DP 3

DP 4

pH 7 .95

7.75

8.10

7 .82Alkalinity 250

230

248

250Specific Cond 1695

1570

1700

1660Hardness (tot) 240 .1

236.4

251.3

259.0Phosphate ppm 0 .12

0.115

0.085

0.075N03 ppm 1 .40

1.40

1.55

1.50S04 ppm 12 .0

10.25

12.7

11 .9Cl pp . 378 .4

350.0

383.5

372.4Ca 58

56

58

58Mg 364

332

288

340Fe 0

0

0

0Cu 0

0

0

0Pb 0

0

0

0Zn 0

0

0

0Cc 0

0

0

0

Table V

Protozoans Found in Exposed Sterile Containers Set Out

in Sandy Point, Abaco Island

ar No .12/29/69

1

02

03

04

05

Cyclidium sp ., Chrysomonad, Holophrya sp .6

07

1 small ciliate, 2 small flagellates8

09

0

12/30/691 0

small ciliate, 2 small flagellates, hypotrich, Chilodonella sp.,Bodo sp .

11

small ciliate12

Spathidium sp ., Bodo sp ., 3 small flagellates, small ciliate13

Cryptomonas sp ., 3 small ciliates, small flagellate14

small flagellates, small ciliates15

small ciliate, Spathidium type16

small flagellate, small ciliate17

small flagellate, Chilodonella sp .18

Oxytricha sp ., small flagellate, small ciliate19

no collection, jar knocked over20

small ciliate, small flagellate, medium-sized ciliate21

022

023

o collection, jar knocked over24

0

Table VI

Surface Water Sample Sites, Approximate Elevation Above Sea Level

3 .5'

5'

± 1/2'3 -4 .75'3 -5

Probably 5 feet or greater, definitely less than 10 feet

21 - Less than 5'22

- Less than 5'23 - Probably 5' or greater - definitely less than 10 feet24 -A$ 5 feet

41 5

416

Table VII

Water Quality in Colonization Study Containers on Day of

Examination for Protozoans

Table VIII

pH Values for Water in Colonization Study Containers on Day

of Examination for Protozoans

19- None taken (container knocked over - water spilled)20- 7 .8,H20 highly colored21- None taken (lizard in container)22- 8 .4923- None taken (container knocked over - water spilled)24- 8.65

Sampler No . 2 10

pH 8.71 8 .46

Alkalinity 295 .0 255

Specific Cond . 555 .0 510 .0

Hardness (tot) 259 .0 217 .6

Phosphate ppm 0 .22 0 .32

N03 ppm 0 .72 0 .68

S04 ppm 6 .0 6 .0

Cl ppm 13 .30 17 .38

Ca 118 104

Mg 208 208

Fe 0 0

Cu 0 0

Zn Trace w 0 .04 ppm 0 .14 ppm

Cr 0 0

Sampler No.

1- 8.572- 8 .71 (retained for further tests ; see Table 7)3- 8 .624- 8 .605- 8 .706- 8.507- 8 .708- 8 .559- 8 .65

10- 8 .46 (retained for further tests ; see Table 7)11- 8 .6012- 8 .4813- None taken (dead mouse in container)14- 8 .5815- 7 .6016- 8 .5517- 8 .4218- 8 .3 (water highly colored by bite of vegetation that had

fallen in)

Tabl

eIX

Species of Protozoa Found in Cultured Soil Samples from Abaco Island, Bahama

s

Station

:

12

3 4

56

78

910

1112

1314

15 1

617

1819

2021

2324

25 26 DPI DP2 DP3 DP4

Phylum Protozoa

Subphylum Plasmodroma

Class Mastigophora

Subclass Phytomastigia

Order Chrysomonadida

Family Chromulinidae

Oikomonas termo (Ehrenberg)

0. sp

.W

M MW

'WFamily Ochromonadidae

Monas socialis Kent

WC

MCM. sp

.CW

CC

WW

WW

Order Cryptomonadida

Family Cryptomonadidae

Cyathomonas truncate Ehrenberg

MW

CW

CW

WM

WW

CWC

MC

Order Phytomonadida

Family Chlamydomonadidae

Chlamydomonas sp

.C

Order Euglenoidida

Family Euglenidae

Euglena viridis Ehrenberg

MFamily Astasiidae

Petalomonas abscissa Stein

wP. anguata (Klebs)

W

P. sp

.1

CP. sp

. 2

WRhabdomonas incurva Fresenius

WC

WW

WMW

W CW

WW

WW

MFamily Anisonemidae

Anisonema emarginatum Stokes

WAnisonema ovale Klebs

CC

CPeranema trichophorum (Ehrenbe

rg)

CHeteronema sp

.MW

Entosiphon sulcatum (Dujardin)

WNotosolenus apocamptus Stokes

CCW

WC

W

Station

:

Subclass Zoomastigia

Order Rhizomastigida

Family Mastigamoebidae

Mastigamoeba

invertens

Klebs

M. sp

.Order Protomonadida

Family Codosigidae

Monosiga~v~r~

Kent

Family

Amphimonadidae

Streptomonas

cordata

(Perty)

Family Bodonidae

Bodo

amoebinus

Lemm

.B

.caudatus

Dujardin

B.celer

Klebs

B. edox Klebs

B.

Rlobosus

Stein

B.minimus

Klebs

B.

obovatus

Lemm

.B

.repens

Klebs

B-uncinatus

(Kent)

Rhynchomonas

nasuta

(Stokes)

Colponema.

loxodes

Stein

Cercobodobodo Lemm.

C.crassicaudaLemm

.C

-sp

.Cercomonas

crassicauda

Dujardin

C.longicauda

D.

Order Polymastigida

Family Tetramitidae

Tetramitus

pyriformis

Klebs

WClass Sarcodina

Subclass Rhizopoda

Order Proteomyxida

Family Vampyrellidae

Unknown

WNuclearia

delicatula

Cienkowski

C

M

1 W

2 C

34 5

67

89 10

1112

1314 W

1516

1718

19 C

2021

23 M

2425 26 DP1 DP2 DP3 DP4

W

M

M

W W

MC

wC

CW C

WC

CCW

CW

MW

MC

WMW

WM

WM

C W

_C

CC

CM

CW M

MV

CC

CW

CC C C

CW

WC CW

CC

WW

M WM

M

M

CW

M

Station

:

1 2 3 4 5 6 7 8 9

10 11 12 13 14 15 16 17 18 19 20 21 23

24

25 2

6 DP

1 DP

2 DP

3 DP

4

Order Amoebida

Family Naegleriidae

Naegleria

gruberi

(Schardinger

)

W

M M W M

MW W MC

W C

W

M M M

MFamily Amoebidae

Amoeba

dubia

Schaeffer

W

A.gorgonia

Penard

W

WW

A.guttula

Dujardin

W

W

W

W

M

W

A.limicola

Rhumbler

M

W

M

A.radiosa

Ehrenberg

W

W

W

W

MC

C

A.vespertilio

Penard

W

W

W CW

W

CW CW

CA. sp

. 1

W

A. sp

. 2

W

CVahlkampfia

limax

Dujardin

W W W

W M M M

W

WHartmanella

sp.

W M M

M M

CW

MSubclass Actinopoda

Order Heliozoa

Family Lithocollidae

Astrodisculus

sp.

WUnknown

W

C

CSubphylum Ciliophora

Class Ciliata

Subclass Holotricha

Order Gymnostomatida

Family Holophryidae

Holophrya

sp.

WLagynophrya

sp.

WUrotricha

near

a ilis

Stokes

MU

.ovata

Kahl

f

MLacrymaria

olor

(Auller)

CW

L. near

minimaKahl

CEnchelys

mutans

Mermod

CW2,

E.variabilis

Sveg

.

MET

sp.

WChaenea

near

vorax

Quennerstedt

CC

.sp

.

WEnchelyodon

fusidens

Kahl

?Trache lophyllum

chilense

Burger

C

WT

.near

fusca

Kahl

WFamily Spathidiidae

Spathidium

near

modestum

Kahl

W

Station

:

1 2 3 4 5 6 7 8 9

10 11 12 13 14 15 16 17 18 19 20 21 23

24

25 2

6 DP

1 DP

2 DP

3 DP

4

Family Frontoniidae

Frontonia

leucas

Ehrenberg

WF. sp

.

WPlatynematum

sociale

(Penard)

WCinetochilum

margaritaceum

Pert

y

CW

C CW C C

C CW

W C

CW C C

Family Pleuronematidae

CyclidiumbrandoniKahl

w

w

C.glaucoma

Milller

W

C

W

W

CMW

MW

MW

MC. sp

. 1

W

WC

CWW

M

W

CW

CW

CM

MCW

MCW

MCW

C.sp

. 2

W

C CW

W

W

CSubclass Spirotricha

Order Heterotrichida

Family Metopidae

Metopus

near

asMuller

M.near

rostratus

Kahl

WFamily Spirostomatidae

Spirostomumambiguum

Ehrenberg

CBlepharismanear

steini

Kahl

WOrder Oligotrichida

Family Halteriidae

Halteria

grandinella

(Muller)

WOrder Hypotrichida

Family Oxytrichidae

Oxytricha

fallox

Stein

w

W0. sp

. 1

CW

WC

W

WW

W

W

C CW

0. sp

. 2

W

CUrosoma

cienkowskii

Kowalewski

WU. sp

.

WKahlia

sp.

W

Uroleptus

sp.

W

Strongylidium

sp. 1

MS. sp

. 2

WStichotricha

sp.

CW

Keronopsis

sp.

W

WW

Holosticha

sp.

W

WW

W

CW

WGgptrostyla

muscorum

Kahl

wOnychodromopsis

flexilis

Stokes

w

Station

:1

2 3

4 5

6 7

89

1011

1213

14

1516

17 1

819 20 21

2324 25 26 DPI DP2 DP3 DP4

Family Amphileptidae

Lionotus near anguilla K

ahl

WL. sp

.Hemiofhrys sp. 1

CH. sp 2

WC W

Family Tracheliidae

`?Dileptus binucleatus Kahl

wW

WC

t D. tenuis(Penard)

WFamily Chlamydodontida

e'Z Chilodonella calkinsi Kahl

wC

CC. uncinata (Ehrenberg)

MMCW

Family Nassulidae

Chilodontopsis pseudonassula

(Penard)

CC

Order Trichostomatida

Family Colpodidae

Colpoda as era Kahl

MMW

MM MW

WM

MM

M MW

MM

M MW MW

M W

C. cucullus Mu7

.ler

MW

MWM

MMW

MCW M MW

M W

MC

MW MW

MMW MWMW

M M MW

M CW C

C-sp

.MW

M W

MC M

MW

M C

MM

MCW

Tillina canalifera Turner

wW

WW

Family Microthoracidae

Leptopharynx sphagnetorum

(Levander)

WCW

W CW

C MW

WCWMC

W CW

WW CW

C MC

WDrepanomonas sp

.W

WOrder Hymenostomatida

Family Tetrahymenidae

Tetrahymena pyriformis (Ehrenb

erg)

MW

Colpidium campylum (Stokes)

MW

Monochilum frontatum Schewiako

ffw

CSaprophilus muscorum Kahl

M;t

S. sp

.M

WPseudoglaucoma sp

.MW

M MC

WM

CMW

M

Station

:

1 2 3 4 5 6 7 8

9 10

11

12 1

3 14

15

16 1

7 18

19

20 2

1 23 24 25 26 DP1 DP2 DP3 DP4

Family Euplotidae

Euplotes

near

carinatus

Stokes

W

C

W

W

W

CW

E.near

charon

Stein

MFamily Aspidiscidae

Aspidisca

sp.

CW

W

CW

MC M MC

CSubclass Peritricha

Order Peritrichida

Family Vorticellidae

Vorticella microstoma

Ehrenberg

W

C

W

M = powdered milk culture

C = modified Chu 14

W = boiled wheat culture

DP = duck pond

Table X

Number of Species of Protozoa Found in Cultured Soil

- indicates no sample taken

M = powdered milk cultureC - modified Chu 14W = boiled wheat culture

habitats, significant reservoirs of fresh water species exist, either inthe soil or in the air . This confirms the findings of MAGUIRE (inpress) . The results of the collections from various surface freshwater habitats indicate that the distributional pattern is not con-sistent. Some of the bodies of water contain fairly substantialnumbers of protozoans (20-30) while others contain only a fewspecies (1-7) .

The limited fresh and partially fresh surface waters of AbacoIsland appear to have a more restricted protozoan biota thanwould be present in comparable bodies of water on the mainland .Unfortunately the observations were not sufficiently extensive toconfirm this. However, an impoverished diatom flora was noted onthe island of Dominica by PATRICK (1967) . PATRICK felt this wasdue to a restricted species pool presumably resulting from isolationfrom potential colonizing species . It would be surprising if thesame situation were not true for the protozoans since comparablespecies transport mechanisms are probably operative for bothgroups. In any case the fresh and slightly brackish waters examined

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Samples at Each Sampling Period

WEEK 2 3Sample Total P

/1 M C W M C W M C W Of Species

1 3 0 12 1 0

9 0 3 5 202 3 1 3 2 4 12 1 3 14 243 4 1 1 2 2

2 3 2 3 134 3 1 0 - - - 1 1 2 75 2 3 4 3 6

9 0 3 10 266 3 5 1 0 5

2 1 1 2 147 2 1 3 - - - 1 2 7 148 3 1 2 - - 4 1 0 5 119 2 0 1 - - - 3 1 1 9

10 2 4 4 2 4

6 2 2 7 2011 7 1 7 3 - 8 1 5 10 2512 2 0 0 - - 0 2 2 1 513 2 0 6 - - 6 1 0 2 1414 2 5 7 - - 10 1 2 9 2415 2 2 1 - - 1 0 3 1 816 3 2 3 - - 1 3 2 2 101.7 2 2 0 - 1 - 2 1 4 10E8 1 3 4 - - 8 0 5 9 197.9 2 0 0 - - 0 2 3 1 1520 5 2 2 - - 2 3 6 2 1421 4 2 7 - - 6 2 3 14 2723 4 3 5 4 3

5 2 1 4 1724 2 1 0 1 0 1 1 2 1 525 0 1 0 0 2 0 0 1 326 3 5 3 0 2

1 2 2 3 16DPI 4 2 3 4 2

2 4 3 4 18DP2 7 2 5 3 6

2 4 1 3 21DP3 1 3 6 2 7

5 4 1 4 20DP4 4 1 0 2 1 3 4 2 6 12

on Abaco produced far fewer species than a comparable effort onsimilar waters in continental North America (about 50 species persampling area would be typical) .

Most of the island surface waters are probably of somewhat simi-lar origin, that is, initially descending as rain and then percolatingthrough the limestone substrate to collect in small surface watersupplies . Most of the so-called surface waters are actually in depres-sions in the limestone . Although there was not sufficient time tocarry out detailed water quality determinations, the water qualitydata collected indicate some variation (mainly in specific conduc-tance, hardness and Cl - concentration) in the depressions con-taining water in the Sandy Point area of Abaco Island . If this istrue, variance in numbers of kinds of species found in surface watersis possibly due to both variations in water quality and invasion rate .

2. Colonization studies - Few protozoans were found in thesterile containers placed around Sandy Point . Ten out of the 24jars had no protozoans at all ; the others only had 6 or fewer species .These species could have been air-borne, present in the vegetation thatfell into the jars, or carried in by organisms such as insects, smalllizards, and small mammals . The low number of species indicateseither a very low invasion rate or an unsatisfactory habitat in thejars although the water used maintained almost the same charac-teristics over the 3 week exposure period as the well water source .

As mentioned previously, surface waters were quite limited onthe island . Sampling was usually restricted to wells and small ponds .Species available for invading new areas would be those air-bornefrom the mainland, those air-borne or animal-transported fromthe island surface waters, and those invading from the soil . Thelimited number found in the colonization studies suggests thateither : (1) species were not available for colonizing new areas, or(2) the water in the jars would not support most protozoans dueto inhibiting heat, light, and water quality characteristics. Thedifferences in number of species found is probably indicative ofvariation in air-borne and soil-borne invasion rate since waterquality remained fairly uniform in all the jars .

3. Soil samples 1-26 did not differ from Duck Pond samples innumbers of species present, i .e ., Duck Pond samples did not havea significantly higher or lower number of species than soil samples .Likewise, type of soil did not always determine the number ofspecies found . Two samples, 23 and 24, were clay soils, but one hada relatively high species number (17) and the other had a lownumber (5) . Generally the highest number of species found in anysoil sample was from the wheat culture, but there was much varia-tion in this . No general statement can be made concerning the

424

change in species number over time in each medium . Kinds ofspecies found will vary depending on the substrate . Several specieswere found only from one location, and whereas the 3 species ofColpoda found were common from soil samples, none were foundfrom Duck Pond samples .

Compared with direct sampling and examination of materialfrom aquatic areas, the cultured soil samples usually yielded agreater total number of species . Also the kinds of species found weredifferent for the two sampling methods . Out of 74 species fromfield investigations and 119 species from the lab cultures, only 20species were common to both studies .

Obviously there are problems in comparing data from the labcultures with data produced by sampling from the natural environ-ment. The three types of cultures used were probably selective forcertain species, while other species were not able to survive or reachhigh densities ; therefore, there were probably many species in suchlow densities that they were not detected . Lab cultures certainly donot indicate what conditions are in the natural state . Under naturalconditions in the soil, species are probably either in an encystedresting stage or are found living in capillary or interstitial water .Since may species from soil samples were not found in the fresh-water environment through direct sampling, that environment mustnot be sufficiently favorable for these species to reach a density highenough for the species to be found . The numerous species found insoil samples indicate that many species are present in the area (morethan are found through direct sampling of aquatic areas) and maybe a `reservoir' group which could invade aquatic areas and becomeestablished if physical, chemical and biological conditions arefavorable .

SUMMARY

The cosmopolitan distribution and invasion of fresh-water pro-tozoans were examined on Abaco Island, Bahamas by comparing :(1) samples from surface waters ; (2) containers of sterile waterexposed to colonization ; and (3) cultured soil samples . The dis-tributional pattern of protozoans in the various surface waters wasnot uniform ; the number of species found varied widely (from 1-32sp.) . Out of 74 species from surface waters and 119 species from thesoil cultures, 20 species were common to both studies . Other speciescommon only to soil samples may act as a reservoir group whichfurnishes colonizing species should a suitable aquatic habitat bepresent. Very few species (range of 0-6 per container) were col-

425

lected from jars of exposed sterile water . The variation in numbersand kinds of species found in surface waters may be due to varia-tions in water quality or variation of invaders, while differences innumber of species found in the containers may indicate some varia-tion in air-borne invasion rate since water quality was essentiallythe same .

ACKNOWLEDGEMENTS

We gratefully acknowledge the assistance of PROFESSOR JOHN M.BATES, Director, Center for Aquatic Biology, Department ofBiology, Eastern Michigan University, Ypsilanti, Michigan 48197,and his staff, SALLY DENNIS, JAMES B. LINCOLN, and JAMES PIETRZAK,Research Biologists . We are also indebted to DR. CHARLES W.REIMER of the Academy of Natural Sciences in Philadelphia,Pennsylvania, for generously furnishing a portion of his algal sam-ples so that we could determine what protozoans if any were asso-ciated with them . The enitre project was made possible by thegenerosity of Mr. E. P . PRESCOTT, Cleveland, Ohio, who madethe fishing camp at Sandy Point available for these studies . Mr.GENE ULRICH and his son RICK were extremely helpful in locatingsampling sites. Last but not least the trip was brightened and madememorable by the cheerfulness and many courtesies as well as thedeeply appreciated help provided by Mr . NAPOLEAN ROBERTS .Although the citizens of the town of Sandy Point did not contributeto the scientific investigation, their hospitality and kindness duringthe Christmas holiday season will not be forgotten .

LITERATURE

BROWN, JR., R . M., LARSON, D. A . & BOLD, H. C . - 1964 - Airborne algae : theirabundance and heterogeneity . Science. 1943 (3606) : 583-585 .

CHU, S . P . - 1942 - The influence of the mineral composition of the medium onthe growth of planktonic algae . I . Methods and culture media . ,3 . Ecol.,30 : 284-325 .

COOKE, W . B . - 1956 - Colonization of artificial bare areas by microorganisms .Bot. Rev . 22 : 613-638 .

DARWIN, CHARLES - 1860 - The Voyage of the Beagle . Footnote on pp. 222 ofDoubleday & Company, Inc . Pocketbook N 16 .

GISLEN, T . - 1948 - Aerial plankton and its conditions of life . Biol. Rev . 23 :109-126 .

HUBER-PESTALOZZI, G. - 1937 - Das Phytoplankton des SUsswassers . THIENEMANNBinnengewisser 16 : 62-72 .

KAHL, A. -1935 - Wimpertiere oder Ciliata (Infusoria) .-In DAHL, F., Die TierweltDeutschlands ., Urtiere oder Protozoa . I . G. Fischer, Jena.

426

MAGUIRE, JR ., B . - 1963 - The passive dispersal of small aquatic organisms andtheir colonization of isolated bodies of water. Ecol . Monogr . 33 : 161-185 .

MAGUIRE, JR ., B. (in press) . Community structure of protozoans and algae withparticular emphasis on recently colonized bodies of water . In CAIRNS, JR .,J . et al . (in prep) . The Structure and Function of Fresh-Water MicrobialCommunities, V .P.I. Press, about 300-400 pages.

MAGUIRE, JR ., B. & BELK, D . - 1967 - Paramecium transport by land snails . J .Protozool . 14 (3) : 445-447 .

PASCHER, A . - 1913-1927 - Flagellates - In Die susswasser-flora Deutschlands,Osterreichs and der Schweiz . G. Fischer, Jena.

PATRICK, R . - 1967 - The effect of invasion rate, species pool, and size of area onthe structure of the diatom community . Proc . Nat . Acad. Sci. 58 (4 :)1335-1342 .

SCHLICHTING, JR ., H. E . - 1961 - Viable species of algae and Protozoa in theatmosphere . Lloydia 24 (2) : 81-88 .

SCHLICHTING, JR., H . E . - 1964 - Meteorological conditions affecting the dispersalof air-borne algae and Protozoa . Lloydia 27 (1) : 64-78 .

(Footnote continued of pag, 413)

C. Associated with bluegreen algae on bottom of well .D. Associated with Oscillatoria sp . on rootlets protruding from well wall .E . Associated with Colathrix sp . on wall above waterline. No protozoans noted

until protozoan-free water was added .F. From intestine of tadpole living in abandoned main well .

2 . Well behind schoolhouse, 12/27/69 .3. In pool of water in coconut tree . 12/27/69 and 12/30/69 .4. Crossing Rock Village Swamp 12/27/69 .5. Duck Pond 12/28/69 .G. Plankton .H. Associated with algae on Chara.I. Yellow scum, bottom of pond .J. Bottom debris of pond .

6. Plankton tow in excavated pond . 12/30/69 .7 . Well one mile outside of town, off the road 12/30/69 .8. Well above airport, one mile off road, sample from red algal filaments,

12/30/69 .

42 7