THE DISPERSAL OF ALGAE AND PROTOZOA

BY SELECTED ODONATA

APPROVED*

Major Professor

/y * ' ••

Minor Professor

FT NP< \

M-ys \1ULI Director of the Department of Biology

Dean of the Graduate School

THE DISPERSAL OF ALGAE AND PROTOZOA

BY SELECTED ODONATA

THESIS

Presented to the Graduate Council of the

North Texas State University in Partial

Fulfillment of the Requirements

For the Degree of

MASTER OF ARTS

By

William M. Parsons, B. A,

Denton, Texas

January, 1966

TABLE OF CONTENTS

Page

LIST OF TABLES iv

LIST OF ILLUSTRATIONS v

Chapter

I. INTRODUCTION 1

Significance of the Problem History of the Problem Statement of Purpose

II. MATERIAL AND METHODS 10

Collection of Insects Preparation of Gross External Washings Washings of Specific Boiy parts

III. RESULTS 17

IV. DISCUSSION 25

Pickup and Deposition of Dissemulee As Related to Odonata Behavior

Internal Transport of Dissemules

V. CONCLUSIONS 51

BIBLIOGRAPHY 33

iii

LIST OP TABLES

Table Page

I. Physical Data, Colorado 15

II. Algae and Protozoa Cultured from Selected 19 Colorado Anisoptera

III. Algae and Protozoa Cultured from Selected 21 Colorado Zygoptera

XV. Percent Frequency of Colorado Algae and 23 Protozoa in Cultures and From Insects. . . .

V. Microorganisms Cultured from Dissected 24 Parts of Libellula auripennis (Hagen). . . .

iv

LIST OF ILLUSTRATIONS

Figure Page

I. Map of Collection Locations. . . . 12a

CHAPTER I

INTRODUCTION

Significance of the Problem

Aquatic ecologists are constantly seeking a better

understanding of the complex interrelationships between

higher aquatic organisms and microscopic forms. One such

relationship that requires further work involves the passive

dispersal of microorganisms across land barriers by insects

and higher animals.

Except for dispersal by stream flow or rain runoff,

the introduction of algae, Protozoa, and other microorganisms

into new or isolated aquatic habitats depends upon some mode

of overcoming land barriers. Known modes of passive, overland

transport include (l) dispersal by wind of suspended dissemules,

(2) dispersal by waterfowl and shore birds, and (3) dispersal

by other animals.

It has been shown that the wind may act as an agent of

disseinule dispersal (l, 5, 6, J, 8, 15/ 16, 18, 19, 22, 23,

28/ 29). As early as 1888 it was shown that waterfowl carry

2

numerous genera of microorganisms on their feet (2, 3# 10,

11, 12, 1J, 14, 16, 21, 25). Viable algae and Protozoa have

also been cultured from the intestinal tracts of selected

waterfowl (l6, 20, 21). Vertebrate animals such as frogs,

toads, salamanders, turtles, raccoons, bears, and others, may

also act as agents of passive dispersal (4, 10, 14, 21, 25,

27).

Knowledge of dispersal mechanisms may have far reaching

implications in the fields of water supply and public health.

The presence of undesirable algae in lakes, reservoirs, and

water supply systems, have contributed to tastes, odors,

coloration, slime formation, and corrosion problems.

Asterionella sp. and Synedra sp. have been reported as in-

hibiting proper floe formation in water clarification (26).

Algae in a raw water supply have induced changes in pH,

alkalinity, hardness, and dissolved oxygen, and these changes

have required additional chemical treatment to purify the

water. Acute and often fatal poisoning of livestock due to

ingestion of water from ponds supporting an algal bloom has

also been reported (9). An awareness of modes of microorgani

dispersal may aid in the construction and maintenance of im-

proved water supply systems.

sm

History of the Problem

The earliest study of aquatic insects as possible trans-

port mechanisms was conducted by W. Migula in 1888 ( 1 7 ) . In

studying a pool thirty centimeters in diameter he found a

single aquatic beetle associated with the algae in the pool.

He concluded that the beetle must have carried the algae to

the pool. He later studied six beetles belonging to three

species from five different habitats, and found twenty-three

species of algae attached or associated with the beetles.

The following genera of algae were noted: Anabaena, Characium,

Synedra, Oscillatoria, Scenedesmus, Navicula, Protococcus,

Cocconeis, Palme11a, Penium, Meridion, Chroococcus, Hapalosiphon,

Fragilaria, and Bncynomena. From the results of his study,

Migula concluded that aquatic insects play a much more signifi-

cant role in dispersal than either water birds or air currents.

In 1910 Scott ( 2 4 ) stated that aquatic beetles and some

of the Hemiptera may be the most efficient agents of transport

because the immature forms develop in the water. During this

time algae and Protozoa may becomi attached to them. Irenee-

Marie in 1938 ( 1 0 ) examined a number of dytiscid beetles and

found members of the genus Closterium in their claws. In

studying a dragonfly of the genus Llbellula, he also observed

4

a number of desmids. Mesikommer, in 19^3, (l6) gave credit

to members of the Anisoptera for dispersing microorganisms

from one body of water to another in a limited locality. He

stated that the dissemules were encysted, but that they may

even be viable in the vegetative state if the dispersal dis-

tance was not too great.

Maguire (13) undertook a more exhaustive study of micro-

organism dispersal by aquatic insects using more refined

techniques. Dragonflies were collected using a .22 caliber

pistol, firing shot type cartridges. The possible contamina-

tion of shot dragonflies by microorganisms when they struck

the ground was apparent. "But", states Maguire, "since the

five or six of them which remained on the leaves where they

were hit carried about the same kinds of organisms, appreciable

contamination seems not to have occurred when they fell."

Maguire collected damselflies by placing them into sterile

vials of filtered pond water with sterile corks. The damsel-

flies had little, if any, chance of being contaminated, and

controls consisted of vials of filtered pond water into which

no insect had been placed. He collected the following aquatic

insectss midges, caddisflies, crane flies, and mosquitoes,

at night by attracting them to a clean lighted sheet, picking

5

them off with sterile forceps, and then placing them in a

vial of filtered pond water. The controls showed that con-

tact between the collecting vial and the sheet did not lead

to significant contamination by microorganisms.

Maguire (14) made a comparative study of passive dis-

persal of microorganisms in Texas and Colorado. Birds and

insects capable of overland transport were collected using

a .22 caliber pistol and dust shot. These were washed with

artificial sterile lake water. Controls consisted of auto-

claved dragonflies which were dropped onto the grass to

simulate their fall to earth after being shot. Twenty-four

dragonflies from the genera Dythemis, Plathemis, Libellula,

Tramea, and Gomphus, were shot and of these twenty showed

positive algal growth. Dragonfly washings were studied

microscopically and the following algae were identified*

Chlarnydomonas, Nannochloris, Chlorococcalian-like alga,

Chlorella, Ankistrodesmus, Phormidium, Lyngbya, and Plectonema,

while the controls were negative. The work of Maguire (12#

15, 1^) definitely established that some aquatic insects

were transport mechanisms for algae and Protozoa.

Purpose

It is apparent that only a minute fraction of the

aquatic species that might serve as transporters of viable

microorganisms have been studied, and that much further

work is ne«ded. Maguire's work (12, 13» 1*0 established

that the Odonata may play an important role in the dispersal

of aquatic microorganisms. This study was designed to (a)

show what dissemules may be carried by selected genera of

Odonata, (b) show where the dissemules may be predominantly

carried on the selected insects, and (c) relate the behavior

of the selected Odonata to frequency of occurrence of micro-

organisms on the insects.

CHAPTER BIBLIOGRAPHY

1. Beger, H., "Beitrage zur Okoligie und Soziologie der Luftlebigen (Atmosphytischen) Kieselalgen," Deutche Botanische Gesellschaft, XL (1927), 585-^07.

2. Darwin, C. R., Origin of Species, New York, Random House, Inc., 1859.

3. de Guerne, J. M., "Sur la dissemination des organismes d'eau douce par lea Palmipedes," Societe de Biologie, VIII (1888), 294-298.

4. Edgren, R. A., M. K. Edgren, and L. H. Tiffany, "Some North American Turtles and Their Episoophytic Algae," Ecology, XXLIV (1953)/ 733-739.

5. Gislen, T., "The Number of Animal Species in Sweden with Remarks of Some Rules of Distribution Especially of the Microfauna," Acta University of Lund, XXXVI (1940), 1-23.

6. ., "Aerial Plankton and Its Conditions of Life," Biological Review, XXIII (1948), 109-126.

7. Huber-Pestalozzi, G., "Das Phytoplankton des Sussvassers," Die Binnengewasser, XVI (1937)/ 62-72.

8. Hudson, C. T., "Presidential Address," Royal Microscopical Society, (1889), 169-179.

9. Ingram, W. M., and G. W. Prescott, "Toxic Freshwater Algae," American Midland Naturalist, LII (1954), 75-87.

10. Irene'e-Marie, Prere, Flore Dismldiale de la Region de Montreal, Laprairie, Canada, 1938.

11. Klingel, <3. C., Inagua, New York, Dodd, Mead, and Co., 1940.

8

12. Maguire, B., "Studies Concerning the Passive Dispersal of Small Aquatic Organisms," unpublished doctoral thesis, Department of Biology, Michigan State University, Lansing, Michigan, 1957*

13. . ' •/ Passive Overland Transport of Small Aquatic Organisms," Ecology, XL (1959), 312.

14. ., "The Passive Dispersal of Small Aquatic Organisms and Their Colonization of Isolated Bodies of Water," Ecological Monographs, XXXIII (1963)# 161-185.

15. Meier, F., and C. A. Lindbergh, "Collecting Micro-organisms from the Arctic Atmosphere," Scientific Monthly, XL (1935)# 5-20.

16. Messikomer, E. L., "Untersuchunger uber die Passive Verbreitung der Algae, " Schv.eizerische Zeitischrift fur Hydrolog ie, IX (19437".

17» Migula, W. A., "Die Vertreitungsweise der Algae," Biologlsches Zentralblatt, VIII (1888), 514-517.

18. Pady, S. M., "Quantitative Studies of Fungus Spores in the Air," Mycologia, XLIX (1957)> 339-353.

19. Pennak, R. W., Freshwater Invertebrates of the United States, New York, Ronald Press, 1953-

20. Proctor, V. M., "Dispersal of Freshwater Algae by Migratory Water Birds," Science, CXXX (1959)» 623-624.

21. Schlichting, H. E., Jr., "The Role of Waterfowl in the Dispersal of Algae," Transactions of the American Microscopical Society, LXXIX (196oT^ T&0-166.

22 . ., "Viable Species of Algae and Protozoa in the Atmosphere," Lloydia, XXIV (1961), 81-88.

23. • » "Meteorological Conditions Affecting the Dispersal of Airborne Algae and Protozoa," Lloydia, XXVII (1964), 63-78.

24. Scott, W., "The Fauna of a Solution Pond/" Proceedings of the Indiana Academy of Science, 1910.

25. Tailing, J. P., "The Element of Chance in Pond Popula-tions," The Naturalist, IV (1951), 157-170.

26. Turre, G. J.. "Algae and other Natural Sources of Tastes and Odors in Water Supplies," Taste and Odor Control in Water Purification, West Virginia Pulp and Paper Co., New York Bulletin, 1955-

27- Vinyard, W. C., "Epizoophytic Algae from MollusV-s, Turtles, and Fish in Oklahoma," Proceedings of the Oklahoma Academy of Science, XXXIV (1953)/ 6^65.

28. Winchell, A. N., and E. R. Miller, "The Dustfall of March 9, 1918," American Journal of Science, XLVT (1918), 599-609.

29 . "The Great Dustfall of March 19, 1920," American Journal of Science, H I (1922), 349-364."

CHAPTER II

MATERIALS AND METHODS

Collection of Insects

Collecting techniques were kept as aseptic as possible

in order to assure that microorgansims washed from insects

were not the result of contamination due to use of a partic-

ular collecting device or undue exposure to the atmosphere

or other objects. Achieving this necessitated sterility of

collecting devices, vials, and the culture medium.

Damselflies resting on aquatic vegetation were grasped

with sterile forceps, carried into the field in ninety-nine

per cent isopropyl alcohol, then placed directly into the

culture medium. Dragonflies were collected with nylon insect

nets washed thoroughly with Tide commercial detergent prior

to collections on each sampling date. Capture was accomplished

while insects rested on vegetation or while they were in flight.

Captured insects were then grasped with sterile forceps and

transferred to a vial of sterile culture medium. Net controls

were prepared by dipping random portions of the net into vials

of culture medium after use in sampling.

10

11

Preparation of Gross External Washings

Dragonflies and damselflies from 'which washings of the

entire insect were made, were collected from ten ponds, all

located in the Rocky Mountains of Colorado (Fig. l). These

ponds will hereafter be referred to as sampling stations I-IX.

Table I shows physical data, dates and tiroes of samples for

each sampling station.

The culture medium used throughout this research was

soil water extract. Schlichting (9) has shown that this

medium is desirable when attempting to grow a wide spectra

of microorganisms. This medium was prepared by mixing one

hundred and fifty grams of loam soil with one liter of dis-

tilled water and allowing to stand overnight. The supernatant

was then refiltered through an autoclaved Millipore Filter

Apparatus (Millipore Filter Corporation, Bedford, Massachusetts),

using an HA type membrane filter with a . 4 5 p o r e size. The

receiving filter flask was fitted with a graduated syringe

dispenser. A final pH of between 6.5 and 7*0 was desired,

and a suitable soil was chosen to yield this value. The pre-

pared extract was then dispensed in sixteen milliliter volumes

into autoclaved, non-absorbent cotton stoppered, thirty-two

dram shell vials. The vials were transported in the field in

12

an iced polyfoara cooler to prevent excessive heating of the

cultures of insect washings. Fox each collecting date, two

vials of culture medium were retained in a plant growth chamber

as controls on sterility. As a result there were thirty-tvo

medium controls for the fifty total insect washings.

Vials containing collected dragonflies and damselflies

were returned to the laboratory within six hours and agitated

on a Vortex Jr. Mixer (Scientific Industries, Inc., Queen's

Village, New York) to dislodge dissemules from the insect.

Sterile forceps were used to withdraw insects from the

vials. Insects were preserved or pinned for identification.

External washings thus obtained were then placed in a plant

growth chamber with a sixteen hour photophase and operating

at a temperature of 22°-26°C.

Two weeks after being placed in the culture chamber,

the cultures (external washings) were examined microscopically

by the drop method (8) to determine if growth had occurred.

Subsequent examinations were made at two to three week intervals

for six weeks. The culture vial was stirred using a Vortex Jr.

Mixer and a sample which consisted of one drop/washing/date,

was taken using a sterile pipette. Three horizontal and three

vertical transects were then examined for each drop sample.

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Algal Identifications at the generic level were made

using Palmer (5)# Prescott (6, J), Smith (10), and Starr (11)/

while Jahn (2), and Kudo (3)# were used to identify Protozoa.

Dragonflies and d&mselflies were identified to species where

possible using Borror and DeLong (l), Needham and Westfall

(4), and Usinger (12).

Washings of Specific Body Parts

Washings of dissected external body parts were made

from ten specimens of Libellula aurlpennis (Burmeister)

collected from a pond near the Sabine River in Orange

County, Texas, on July 25 and August 5/ 1965- Each body

part was detached using microdissection scissors soaked in

ninety-nine per cent isopropyl alcohol. The head, legs,

and wings were placed in separate labeled vials of sterile

soil water extract. The abdomen and thorax of each insect

were placed in empty labeled vials, packed in ice, and re-

turned to the laboratory.

The fore- and midgut, and hindgut, were excised from

the thorax and abdomen using aseptic instruments. The

contents were placed in separate labeled vials of culture

medium. The abdomen and thorax were then placed in separate

vials of soil water extract.

CHAPTER BIBLIOGRAPHY

1. Borror, D. J. and D. M. DeLong, An Introduction to the Study of Insects, New York, Holt, Rinehart, and Winston Co., 1964.

2. Jahn, T. L., How to Know the Protozoa, Dubuque, Iowa, Wm. C. Brown Co., -19857

3. Kudo, R. R., Protozoology, Springfield, Illinois, Charles C. Thomas Inc., 1954.

4. Needham, J. G., and M. Westfall, Jr., Dragonflies of North America, Berkeley, California, University of California Press, 1955*

5. Palmer, C. M., Algae in Water Supplies, U. S. Public Health Publication No. 657' 1959.

6. Prescott, G. W,, Algae of the Western Great Lakes Area, Dubuque, Iova, Wm. C. Brown Co., 1964.

7. ., How to Know the Freshwater Dubuque# Iowa, Wm. C. Brown CoTi1964.

8. Schlichting, H. E., Jr., "A Modified Method for the Quantitative Analysis of Phytoplankton Samples by the Drop Method," unpublished paper, Department of Biology, University of Washington, Seattle, Washington, 1954.

9 . • i "The Role of Waterfowl in the Dispersal of Algae, Lloydla, LXXIV (i960), 160-166.

10. Smith, G. M., Freshwater Algae of the United States, New York, McGraw Hill Books, 1950.

11. Starr, R. C., A Comparative Study of Chlorococcum Meneghini and other Spherical, Zoospore-Producing Qenera of the Chlorococcales, Bloomington, Indiana, University of Indiana Press, 1955•

15

16

12. Usinger/ R. L., Aquatic Insects of California, Berkeley, California, University of California Press, 1965-

CHAPTER III

RESULTS

External washings of thirty-three dragonflies belonging

to six species and fifty-three damselflies belonging to five

species produced a total of fifty positive cultures. Data

are shown in Tables II and III.

The dragonflies Aeshna palmata (Hagen) and Erythemis

collocata (Hagen) were found to be carriers of sixteen and

thirteen genera of algae and Protozoa, respectively. Sympetrun.

fasciatum (Walker), atripes (Hagen), Libellula _sat_urat_a

(Uhler), and Tarnet rum corruptum (Hagen), carried seven,

five, four, and two genera, respectively.

Thirty-three specimens of the damselfly Enallagma

cyathigerum (Hagen) carried ten genera of microorganisms.

Six genera of algae and Protozoa were cultured from nine

specimens of Lestes unquiculatus (Hagen). Enallagma civile

(Hagen), Coenagrion sp. (Kirby), and Enallagma sp. ( Charpentier),

carried lesser numbers of genera.

In most cases sample size for a given insect species

appeared to greatly influence the number of different genera

17

18

of algae and Protozoa that were shown being carried by that

species. Where the sample number for a species was small,

fewer genera of microorganisms were cultured in the washings.

Pleuronema sp. was identified in one net control culture

planted August 15/ 1964. This protozoan could have come from

the net or from a previously collected insect. No other micro-

organisms were observed in either net or medium controls at

any examination time.

Frequency of occurrence for each alga and protozoan is

shown in Table IV. These data are separated according to

per cent from a total of fifty cultures and from eleven insect-

species collected.

Chlorococcum sp. appeared in twenty-six per cent of the

cultures and from seventy-two per cent of the insects collected,

while Chlorella sp. appeared in twenty-two per cent of the

cultures and from forty-five per cent of the insects.

The blue-green alga Phormidium sp. was found in sixteen

per cent of the cultures and from forty-five per cent of the

collected insects. Euglenoids were found at relatively low

frequencies on the insect species checked during this study.

The diatom Navicula sp. appeared in six per cent of the cultures

and from nineteen per cent of the insects. Actinosphaerium sp.

19

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was found in twenty-two per cent of the cultures and from

fifty-five per oamt of the insect species, while Amoeba sp. i

appeared in sixteen per cent of the cultures and from thirty-

six per cent of the insects. Other protozoans were found at

lesser frequencies.

These results indicate that filamentous Cyanophyta

were not carried with as great a frequency as the unicellular

Chlorophyta. Fhormidlum sp. was an exception, its having the

same frequency from insect washings as Ghlorella sp.

The results from washings of dissected external body

parts and alimentary tracts are given in Table V. The leg

and wing washings contained ten and eight genera respectively,

more than any other parts washed. No algae or Protozoa were

found in the alimentary tract of Libellula auripermis, although

fungal spores were present. Only one net control of a series

of ten showed evidences of contamination, in which Navicula sp.

was identified after four weeks of culturing. This would in-

dicate that (l) contamination occurred during dissection, (2)

that Navicula sp. came from a previously collected insect, (3)

that the diatom came from the net, or (4) that contamination

occurred during examination procedures.

TABLE IV 23

PER CENT FREQUENCY OF COLORADO ALGAE AND PROTOZOA OCCURRING

IN FIFTY CULTURES AND FROM ELEVEN INSECT GENERA

Organism % Frequency in Cultures

% Frequency from insects

Division Cyanophyta Anabaena jya. Arthrospira sp. Aulosira sp. Calothrix Chroococcus sp. Nostoc sp. Phormidium sj

Division Chlorophyta Chlaroydontonaa sp. Chlorella sp. Chlorococcum sp. Cosmarium sp. Nannochloris ®£* Protococcus sp. Scenedesmus Spirulina sp. Stichococcus S£. Ulothrix sp.

Division Euglenophyta Euglena sp. Euglena-like

Division Chrysophyta Gomphonema sp. Navicula S£.

Phylum Protozoa Agtlnoaphaerlum sp. Actlnophrys ap. Amoeba sp.

8 2 2 2

14 12 16

6 22 26

2 2

10 6 2 4 6

4 12

2 6

22 4

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9 . 1 1 8 . 9

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36.2

TABLE V

MICROORGANISMS CULTURED FROM DISSECTED

PARTS OF LIBELLULA AURIPEMNIS (HAOEM)

24

Structures Algae and Protoaoa

EXTERNAL

Abdomen Anabaena sp., Phormidium sp.

Thorax Chroococcus sp., Navicula sp.

Legs Chlorella sp., Anabaena sp., Chlorococcum sp., Dictyo-sphaerium sp., unclassified encysted Protozoa, Hormidiop-sis sp., Lyngbya sp., Oocystis sp., Protococcus sp., Spongio-chloris sp.

Head Unclassified fungal spores, Chlamydomonas sp., Protococcus sp., Chlorella sp.

Wings Unidentified fungal spores, Chlamydomonas sp., Chlorella sp., Chroococcus sp., Meristnopedia sp., Scenedesmus sp., Synedra sp.

INTERNAL

Fore- and midgut

No organisms discerned

Hindgut Unclassified fungal spores

CHAPTER IV

DISCUSSION

Although the collecting net was detergent washed prior

to use in the field, random washings of the net into culture

media were made as controls in order to elucidate the possible

contamination by microorganisms from the net. When dragon-

flies resting on twigs or branches close to the pond or lake

surface were collected, care was taken so the net made no

contact with the water. Possible contamination of the net

could be minimized by the use of a light monofilament, easily

cleaned, fast drying net. Secondly, the use of a number of

nets, each washed and wrapped in cellophane or plastic, could

be used to collect dragonflies, using one net per insect.

When the problem of microorganism dispersal by selected

Odonata was formulated, it was thought that the use of a

medium with a capability of growing a broad spectrum of

microorganisms should be used, and for this reason soil water

extract was chosen. Schlichting (5) has shown that this medium

will grow more species of algae and Protozoa than any other

single medium.

25

26

Due to the short interval of time in which populations

of some microorganisms rise and decline, especially the

Protozoa, various microorganisms could have been overlooked

as a result of the time intervals between examination of

cultures. However, Schlichting (3) has shown that the decline

for Protozoan populations does not usually occur until after

a two week duration.

Pickup and Deposition of Dissemules As

Related to Odonata Behavior

Resting Habits

Many Odonata prefer to rest on damp soil or twigs

during periods of inactivity. Needham and Westfall (1)

refer to Libellula luctuosa (Hagen) as "resting occasionally

on reed tips and hanging by their feet inches above the water."

Both Sympetrum atripes (Hagen) and Sympetrum fasciatum

(Walker) were collected while they were resting on damp soil.

Libellula saturata (Uhler) and Tarnetrum corruptum (Hagen)

were collected on damp twigs, as were Enallagma cyathigerum

(Charpentier) and Lestes ungulculatus (Hagen). All carried

algae which may exist in a damp, terrestrial environment.

27

Srythemls collocata (Hagen) was also observed resting on damp

soil and twigs, vhich possibly accounted for the appearance

of terrestrial algae such as Protococcas sp., Chlorococcum sp.,

and Phormidlum sp. The greater number of genera in cultures

from washings of _L. auripennis (Hagen) has indicated that

contact between legs and damp soil or twigs may effect dis-

semulization* In addition, resting on emergent twigs short

distances above the water may hare increased the chances of

being "splashed" by waves and therefore resulting in retention

of algae and Protosoa on the insect.

Feedings Habits

Dragonflies and dnmselflies are carniverous insects,

and may prey upon gnats or mosquitoes in flight, or obtain

their food by catching neustonic insects on the surface of

the water* A dragonfly or damselfly which feeds upon these

neustonic insects makes frequent contact with the water,

thereby effecting pickup and deposition of microorganisms.

The behavior of Aeshna palmata (Hagen) closely resembles

that described in the previous paragraph* This insect makes

frequent contact with the surface of a pond while feeding.

Microorganisms may be retained in the water droplets remain-

ing on the insect after splashing the water.

28

Oviposition Habits

Modes of oviposition of female dragonflies and damsel-

flies may aid in explaining microorganism pickup and depo-

sition. The female A. palmata deposits eggs on the submerged

portion of emergent reeds and grasses. Cyclotella sp.,

Gomphonema sp., Cymbella sp., and Navicula sp., were washed

from this insect. These are usually epiphytic, and could

adhere to the legs, thorax, or abdomen of the insect as it

leaves the water. Both Sympetrum atripes and fasciatun

oviposit by submerging their abdomens and depositing egqs

directly into the water or on the submerged portions of

emergent vegetation, thus assuring contact with aquatic-

microorganisms. Observations of ovipositing Erythemis

collocata (Hagen) revealed that the female contacts the

surface of the water at intervals of six to eight feet.

This behavior may account for the appearance of aquatic

algae and Protozoa.

The adults of the genus Lestes frequent the margins of

ponds, bogs, marshes, and other areas where there is emergent

vegetation. Although these were not observed ovipositing,

Usinger ( 5 ) states that members of this genus "oviposit above

the water in standing plants such as Typha, Sciripus, Sparganium,

29

willows, and grasses.." Splashing due to waves, or contact

with damp soil, rocks or twigs, could account for the presence

of algae and Protozoa in the cultures of washings of L.

unguiculatus.

Internal Transport of Dissemules

Culturing of the gut contents of auripennis indicated

that viable algae and Protozoa may not be carried internally

in a viable state by this species. However, studies by

Stewart and Schlichting (4) have shown the presence of highly

resistant fungal spores and of Nostoc sp. cultured from the

gut contents of Gomphus externis (Hagen). Additional cultures

of gut contents of other dragonfly species are needed to

further elucidate this aspect of the problem.

Significance of Odonata Migration

That aquatic insects do migrate has been established by

Pennak (2) who has stated that, "adult aquatic insects capable

of flight easily migrate overland, often in what appears to be

a completely random fashion, and come to occupy suitable new

areas." He goes on to say, "anyone who follows the biological

development of a new, man-made reservoir is bound to be im-

pressed with the promptness with which the reservoir becomes

colonized with aquatic animals."

50

Dragonfly migrations have been reported from Germany,

East Prussia, Egypt, Sudan, Tanganyika, Uganda, Southwest

Africa, British Honduras, Danzig, and China (6). Libellula

quadrimaculata (Linnaeus) was observed in North Wales "literally

in thousands, and appeared to be flying slowly eastward from

11 AM to 4 PM." (6). A report from Kenya stated "for two

days since October 2, or possibly earlier, we have had a

constant stream of dragonflies coming over the house from the

direction of Lake Victoria," (6). The species were later

identified as Pantala flavescens (Fabricius).

The nature and extent of dragonfly and damselfly migration

has been limited to scattered reported observations. More re-

fined techniques have been, and are being, sought in order to

further elucidate this problem. This knowledge is of impor-

tance in explaining geographical distribution- of algae and

Protozoa in isolated bodies of water.

CHAPTER V

CONCLUSIONS

1. Data presented established that aquatic insects

sampled carry viable dissemules of algae and Protozoa.

2. The validity of results obtained depend on the use

of aseptic methods of collection, culturing techniques, and

the examination of the cultured washings.

J. Although soil water extract is the best single

medium for growing algae and Protozoa, it may have restricted

the growth of some dissemules.

4. Some microorganisms may have been overlooked due

to the examination time interval.

5. Results indicate that the legs and wings of the

Odonata sampled play a major role in pickup and deposition

of dissemules.

6. Pickup of dissemules probably occurs while on damp

•oil or twigs, or on emergent twigs close to the surface of

the water.

31

32

7. Frequent contacts with water during feeding or

oviposition may result in pickup or deposition of micro-

organi sms.

8. Data indicated that algae and Protozoa were not

transported in a viable state in the gut of the ten speci-

mens of L. auripennis sampled.

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