Chapter 6

Flatworms: Turbellarians and Nemertea

Jurek Kolasa Seth Tyler
Department of Biology, McMaster School of Biology and Ecology, University of
University, Hamilton, Ontario Canada Maine, Orono, Maine USA

I. Introduction to Turbellaria and Nemertea IX. Collection, Culturing, and Preservation

II. Turbellaria: Introduction and Status in the Animal X. Taxonomic Key to Species of Freshwater
Kingdom Nemerteans
III. Anatomy and Physiology of Turbellarians XI. Selected References
A. General External and Internal Anatomical
Features
B. Environmental Physiology
IV. Ecology of Turbellarians I. Introduction to Turbellaria
A. Life History
and Nemertea
B. Distribution
C. Behavioral Ecology More than 200 species of turbellarians and 2–3 species of
D. Foraging Relationships, Predators, and nemerteans live in freshwaters of North America. Even
Parasites though they are important predators in freshwater ecosystems
E. Population Regulation and Density and are often common in relatively high numbers, their ecol-
F. Functional Role in the Ecosystem ogy and systematics have been less studied than that of many
V. Collecting, Rearing, and Identification Techniques other common aquatic invertebrates. This limited attention
for Turbellarians is partly due to difficulty in identifying and preserving such
VI. Identification of North American Genera of soft-bodied animals. Most turbellarians become unrecogniza-
Turbellarians ble in routinely preserved field samples. Therefore, the study
A. Taxonomic Key to Orders and Suborders of of live specimens, often in conjunction with histological
Turbellarians study of individually processed specimens, is the only reli-
B. Taxonomic Key to Catenulida able means for identifying species. Accurate identification
C. Taxonomic Key to Macrostomida of nemerteans also requires histological study. Considering
D. Taxonomic Key to Lecithoepitheliata their relative obscurity, many interesting discoveries lie
E. Taxonomic Key to Prolecithophora, Proseriata, ahead concerning the reproductive biology, dispersal, endo-
and Tricladida symbiosis, and community structure of these worms.
F. Taxonomic Key to Typhloplanoida
G. Taxonomic Key to Kalyptorhynchia and Similar
Forms II. Turbellaria: introduction
H. Taxonomic Key to Dalyellioida
VII. Nemertea: General Characteristics and External
and status in the animal Kingdom
and Internal Anatomical Features The turbellarian flatworms are basal among the Bilateria,
VIII. Ecology of Nemertea showing such presumably primitive features as an incom-
A. Life History and General Ecology plete gut—with only a mouth and no anus—and an acoe-
B. Physiological Adaptations lomate body structure[79]. Most turbellarians are members
C. Behavioral Ecology of the phylum Platyhelminthes, which also comprises
D. Functional Role in the Ecosystem the major parasitic groups of flatworms—namely the
Monogenea (ectoparasitic flukes), Cestoidea (tapeworms),

Ecology and Classification of North American Freshwater Invertebrates

Copyright © 2010,
2010 Elsevier Inc. All rights reserved. 143
144 Ecology and Classification of North American Freshwater Invertebrates

Table 6.1 List of North American species of microturbellarians. The arrangement of orders and families follows the
taxonomic listing of Tyler et al.[98]; see also Lanfranchi and Papi[57].
Catenulida

Family Catenulidae Stenostomum temporaneum Kolasa 1981

Catenula confusa Nuttycombe, 1956 Stenostomum tuberculosum Nuttycombe & Waters, 1938

Catenula lemnae Duges, 1832 Stenostomum uronephrium Nuttycombe, 1931

Catenula leptocephala Nuttycombe, 1956 Stenostomum ventronephrium Nuttycombe, 1932

Catenula sekerai de Beauchamp, 1919 Stenostomum virginianum Nuttycombe, 1931 (probably

synonymous with S. sphagnetorum)
Catenula virginia Kepner & Carter, 1930
Macrostomida
Catenula turgida (Zacharias, 1902)
Family Macrostomidae
Family Chordariidae
Macrostomum appendiculatum Fabricius, 1826
Chordarium europaeum Schwank, 1980
Macrostomum collistylum Ferguson, 1939
Family Stenostomidae
Macrostomum curvituba Luther, 1947
Myostenostomum tauricum (Nasonov, 1924)
Macrostomum gilberti Ferguson, 1939
Rhynchoscolex platypus Marcus, 1945
Macrostomum glochistylum Ferguson, 1939
Rhynchoscolex simplex Leidy, 1851
Macrostomum lewisi Ferguson, 1939
Rhynchoscolex sp. 1
Macrostomum norfolkensis Jones & Ferguson, 1940
Rhynchoscolex sp. 2
Macrostomum ontarioense Ferguson, 1943
Stenostomum anops Nuttycombe & Waters, 1938
Macrostomum orthostylum Braun, 1885
Stenostomum anatirostrum Marcus, 1945
Macrostomum phillipsi Ferguson & Stirewalt, 1938
Stenostomum arevaloi Gieysztor, 1931
Macrostomum reynoldsi Ferguson, 1939
Stenostomum beauchampi Papi, 1967
Macrostomum riedeli Ferguson, 1939
Stenostomum brevipharyngium Kepner & Carter, 1931
Macrostomum ruebushi Ferguson, 1940
Stenostomum ciliatum Kepner & Carter, 1931
Macrostomum sensitivum Silliman, 1884
Stenostomum cryptops Nuttycombe & Waters, 1935
Macrostomum sp. 1
Stenostomum glandulosum Kepner & Carter, 1931
Macrostomum sp. 2
Stenostomum grande Child, 1902
Macrostomum sp. 3
Stenostomum kepneri Nuttycombe & Waters, 1938
Macrostomum tennesseensis Ferguson, 1939
Stenostomum leucops (Duges, 1828)
Macrostomum tuba Graff, 1882
Stenostomum mandibulatum Kepner & Carter, 1931
Macrostomum virginianum Ferguson, 1937
Stenostomum membranosum Kepner & Carter, 1931
Family Microstomidae
Stenostomum occultum Kolasa, 1971
Microstomum lineare (Müller OF, 1774)
Stenostomum pegephilum Nuttycombe & Waters, 1938
Lecithoepitheliata
Stenostomum predatorium Kepner & Carter, 1931
Family Prorhynchidae
Stenostomum pseudoacetabulum Nuttycombe & Waters,
1935 Geocentrophora cavernicola Carpenter, 1970

Stenostomum simplex Kepner & Carter, 1931 Geocentrophora sphyrocephala de Man, 1876

Stenostomum sphagnetorum Papi in Luther, 1960 Prorhynchus stagnalis Schultze M, 1851

Chapter | 6 Flatworms: Turbellarians and Nemertea
Table 6.1 Continued
Prolecithophora
Family Plagiostomidae
Hydrolimax grisea Haldeman, 1843
Bothrioplanida
Family Bothrioplanidae
Bothrioplana semperi Braun, 1881
Proseriata
Family Otomesostomidae
Otomesostoma auditivum (Du Plessis, 1874)
Dalyellioida
Family Provorticidae
Genus indet. species indet.
Provortex virginiensis Ruebush & Hayes, 1939
Family Dalyelliidae
Castrella graffi Hayes WJ, 1945
Castrella pinguis (Silliman, 1884)
Dalyellia viridis (Shaw, 1791)
Microdalyellia abursalis (Ruebush, 1937)
Microdalyellia circobursalis (Ruebush, 1937)
Microdalyellia fairchildi (Graff, 1911)
Microdalyellia gilesi Jones & Hayes, 1941
Microdalyellia rheesi (Graff, 1911)
Microdalyellia rochesteriana (Graff, 1911)
and Trematoda (endoparasitic flukes)—and which is con-
sidered part of the major Bilaterian group Lophotrochozoa.
Acoel turbellarians, a few of which are inhabitants of
freshwater, were once considered part of this phylum but
are now separated in their own phylum, the Acoelomorpha,
outside the Lophotrochozoa and at the very base of the
Bilateria, by virtue of their placement on phylogenetic
trees constructed from sequence data in certain genes[1].
The class Turbellaria is, therefore, paraphyletic, if not
polyphyletic, and so is no longer considered a valid taxon.
Recent evidence from genomics and studies of stem cells
shows the flatworms to have arisen from a common ances-
tor at the base of the Bilateria, but to be paraphyletic.
The turbellarian platyhelminths are ranked in sev-
eral orders, most of which have members in freshwaters
(Table 6.1, Fig. 6.1). The Catenulida is the most basal
145
Microdalyellia rossi (Graff, 1911)
Microdalyellia ruebushi Luther, 1955
Microdalyellia schockaerti Willems, Artois, Jocque, &
Brendonck, 2007
Microdalyellia sillimani (Graff, 1911)
Microdalyellia tennesseensis (Ruebush &
Hayes, 1939)
Microdalyellia virginiana (Ruebush, 1937)
Typhloplanida
Family Typhloplanidae
Amphibolella segnis Findenegg, 1924
Ascophora elegantissima Findenegg, 1924
Bothromesostoma personatum (Schmidt, 1848)
Castrada hofmanni Braun, 1885
Castrada lutheri Kepner, Stirewalt &
Ferguson, 1939
Castrada sp. indet.
Krumbachia cf. hiemalis Schwank, 1979
Krumbachia minuta Ruebush, 1938
Krumbachia virginiana (Kepner & Carter, 1931)
Limnoruanis romanae Kolasa, 1977
Mesostoma arctica Hyman, 1938
Mesostoma columbianum Hyman, 1939
Mesostoma craci Schmidt, 1858
and, although its relation to other turbellarians is not at
all clear, it is provisionally considered sister group to the
remaining platyhelminths, which together constitute the
Rhabditophora. Orders of rhabditophorans are rather dis-
tinctive, but relationships among them are still under
debate[58]. For practical purposes, turbellarians are often
distinguished also according to size into microturbellar-
ians (less than a few millimeters) and macroturbellarians
(on the order of centimeters in length). This division does
not reflect phylogenetic relationships. Macroturbellarians
in freshwater habitats are planarians; in marine habitats
this category also includes polyclads; all other turbellar-
ians are microturbellarians. Species reaching lengths of
about 1 cm are also known among groups otherwise con-
sidered microturbellarians, including the Hofsteniidae
(Acoela), the Prolecithophora, and the Lecithoepitheliata.
146 Ecology and Classification of North American Freshwater Invertebrates

Figure 6.1 Schematic representation of higher turbellarian taxa found in freshwaters with an emphasis on the most useful diagnostic features:
(A) Acoela (one unidentified species); (B) Catenulida (e.g., Catenula, Stenostomum, I and II represent two successive zooids); (C) Macrostomida
(Macrostomum, Microstomum); (D) Lecithoepitheliata (e.g., Prorhynchus, Geocentrophora); (E) Dalyellioida (Gieysztoria, Microdalyellia);
(F) Typhloplanoida (e.g., Mesostoma, Olisthanella, Castrada); (G) Kalyptorhynchia (e.g., Gyratrix; intestine omitted for clarity); (H) Bothrioplanida
and Tricladida (e.g., Bothrioplana, Dendrocoelopsis). Co, copulatory organ; cp, ciliated pits; i, intestine, m, mouth; n, protonephridial duct; o, ovary;
ph, pharynx; pr, proboscis; s, stylet, st, statocyst; t, testes; v, vacuolized tissue; y, yolk glands.

The ­ largest macroturbellarians are planarians from Lake

Baikal, Siberia, reaching lengths of over half a meter[96].
The total number of known freshwater turbellarians is
estimated to be about 1300[85]. Of the approximately 500
well-established species of freshwater microturbellarians,
about 400 have been recorded in Europe (350 listed by
Lanfranchi and Papi[57]), and approximately 150 occur in
North America (Table 6.1), many of which are the same as
those found in Europe. Many species and genera of micro-
turbellarians remain to be discovered in North America.
Records from other regions of the world are sparse. Africa,
South America, and Papua New Guinea trail closely
behind North America in the number of species, but such
numbers are not likely to be truly representative of these
still little-studied regions. Differences in species richness
more likely reflect research effort, and even the number of
species in North America must be highly underestimated.
Triclads (often called planarians) are the more thoroughly
studied; the number of recorded planarian species in North
America is approximately 40, but the actual number of
taxa is likely to be substantially higher.
III. Anatomy and physiology of
turbellarians
A. General External and Internal Anatomical
Features
Most freshwater microturbellarians are less than 1 mm
in length, although some can reach several millimeters.
Triclads, constituting the freshwater macroturbellarian
group, are distinctly larger, with most species in the range
of one to several centimeters long (Fig. 6.2.)
The turbellarian body is elongated, lacking hard skeletal
elements, and usually tapered at the ends. Sometimes lat-
eral flaps near the cerebral region or a short tail-like ­section
are present. The macroturbellarians like triclads are flat
B
C
A
Figure 6.2 Examples of different types of heterocellular female
gonads: (A) Typhloplana, Dalyellia; (B) Bothrioplana; (C) Prorhynchus,
Geocentrophora.
and strap-shaped, as the name Platyhelminthes implies, but
most flatworms are not, being instead cylindrical, circular
in cross section, with relatively little differentiation of the
dorsal and ventral surfaces. Some species that reproduce
asexually may be chain-like in shape, composed of several
cylindrical zooids (Figs. 6.3b, 6.4a,b, 6.12a). Turbellarians
may be colorless, white, red, bluish, green, black, brown,
or yellowish, depending on epidermal and parenchymal
pigments, gut contents, and symbiotic algae. Some species,
such as Dugesia tigrina and Hydrolimax grisea, develop
characteristic patchy patterns of pigments (Figs. 6.5–6.9.)
Anatomically, the most prominent turbellarian features
are a ciliated epidermis with sometimes prominent rhadiform
secretions from subepidermal glands protruding through,
an intestine without anus, a ventral mouth, and a complex
reproductive system (Figs. 6.1, 6.9, 6.12d). The ciliated
epidermis covers the entire body surface. The rhabdiform
Chapter | 6 Flatworms: Turbellarians and Nemertea
Figure 6.3 Some representatives of Catenulida: (A) Catenula turgida;
(B) Catenula lemnae; (C) Catenula leptocephala; (D,E) Rhynchoscolex
simplex. Length of species ranges between 0.35 and 5.00 mm. B, brain;
f, furrow; i, intestine; m, mouth; no, protonephridial duct; oc, ovary; ph,
pharynx; st, statocyst.
Figure 6.4 Representatives of Catenulida: (A) Stenostomum leu-
cops; (B) S. beauchampi; (C) S. glandulosum; (D) S. brevipharyngium;
(E) Myostenostomum tauricum. B, brain; cp, ciliated sensory pits; g, mus-
cular ring of intestine or gizzard; i, intestine; lo, light-refracting organs; m,
mouth; n, protonephridial duct; pg, pharyngeal glands; ph, pharynx; tc, tac-
tile cilia.
secretions are light-refracting structures known as rhabdoids
and arise in glands that are typically positioned beneath the
epidermis (adenal rhabdoids); in some cases, the glands
may lie within the base of the epidermis (dermal rhab-
doids). It is often difficult to distinguish dermal from ade-
nal rhabdoids in live specimens, particularly because even
rhabdoids from subepidermal glands may be prominent
147
Figure 6.5 Representatives of Macrostomida and copulatory organs
of various families: (A) Macrostomum tuba; (B) Microstomum lineare;
(C) copulatory organ of M. lineare; (D) stylet of Macrostomum gil-
berti; (E) stylet of an unidentified Macrostomum from New Haven, CT;
(F) sclerotized copulatory apparatus of Microdalyellia rossi
(Dalyelliidae); (G) sclerotized copulatory apparatus of M. tennesseensis;
(H) copulatory organ of Opistomum pallidum containing a spiny revers-
ible cirrus (Typhloplanidae). Ci, cirrus; co, copulatory organ; cp, ciliated
pits; e, eye; i, intestine; m, mouth; o, ovary; ov, egg; ph, pharynx; ps, pro-
static secretions; s, sperm; t, testes; vs, seminal vesicle.
Figure 6.6 Schematic representation of the major anatomic features
used in taxonomy of Tricladida. Ad, adenodactyl; b, bursa; bd, bursal
duct; ca, common genital atrium; im, internal muscle layer of the pharynx
(two possible types shown); m, mouth; ma, male atrium; odc, common
oviduct; ph, pharynx; pb, penis bulb; pp, penis papilla; sd, sperm duct; v,
vagina; vs, seminal vesicle.
only where they sit in gland necks within the epidermis.
Adenal rhabdoids are most abundant among frontal glands
that open onto the anterior tip of the body; and in their
course toward the tip, they form bundles known as rod
tracts. Wrona[101] reviewed hypotheses on the role of rhab-
doids in triclads and has provided experimental evidence
that rhabdoids contribute to mucus production. Rhabdoids
are also poisonous and used in prey immobilization or in
deterring predators (Figs. 6.10 and 6.11.)
Beneath the epidermis are layers of body-wall mus-
culature, comprising longitudinal, circular, and diagonal
fibers. Dorsoventral muscle fibers reaching through the
parenchyma are also common, especially in those flat-
worms with flattened body shape. The parenchyma is a
loose tissue filling the body cavity and containing various
cell types, including secretory, excretory, reproductive, and
148 Ecology and Classification of North American Freshwater Invertebrates

stem cells[42,81]. In larger turbellarians, the parenchymal
cells are usually relatively compact, while those in smaller
forms can be quite loose and vacuolated. Turbellarians
possess numerous glands, of which the frontal, pharyn-
geal, shell-producing, and rhabdoid-producing glands are
most common (Figs. 6.12 and 6.13.)
A vast array of sensory receptors enables turbellarians
to react to environmental stimuli. Most are scattered over
the body surface as individual receptors—including mech-
ano-, chemo-, and photoreceptors—but some are organized
into distinct sensory complexes comprising eyes, statocysts,
and chemosensory pits of various kinds. Ciliated pits, in lat-
eral depressions situated in front of the brain ganglion, are
likely to be chemoreceptive. Some structures associated
with these pits, such as the refractive bodies of the catenulid
Stenostomum, have been implicated in phototaxis[64,83], but
photoreceptors have not been definitively identified in cat-
enulids[74]. Other turbellarians have pigment-cup occelli as
eyes, typically with rhabdomeric photoreceptors but some-
times ciliary photoreceptors[81]. The number of eyes is vari-
able, but many species have just one pair. Thigmotaxis is
common and plays a role in a range of behaviors such as
choosing substrate, hunting prey, and avoiding predators.
Among North American freshwater turbellarians, a statocyst
is present only in Catenula, Rhynchoscolex, Otomesostoma,
and an unidentified representative of Acoela[56,93].
Swimming and gliding in the turbellarians is pow-
ered by cilia of the epidermis, and the animals steer these
movements by musculature of the body. Smaller forms can
swim as well as glide on submerged objects, while heavier
forms are restricted to gliding only. Creeping, especially
by macroturbellarians, sometimes in leech-like fashion, is
powered by fast muscular contractions. Adhesive organs
in the form of pseudosuckers or papillae are often used by
many turbellarians to resist water currents and other dis-
lodgments and to retreat from noxious stimuli.

Figure 6.7 Representatives of Tricladida: (A) Hymanella reten- B. Environmental Physiology

uova; (B) Phagocata velata; (C) Dugesia tigrina; (D) D. dorotocephala;
(E) D. polychroa; (F) Procotyla fluviatilis; (G) Polycelis coronata (A and Gaseous exchange in turbellarians occurs through their
B modified from Ball et al.[110]). body walls. Such exchange depends on the surface/volume

Figure 6.8 Representatives of: (A) microturbellarian Seriata (Bothrioplana semperi); (B) Typhloplanoida (Mesostoma craci); and (C–F)
Kalyptorhynchia; (C) Gyratrix hermaphroditus and (D) its stylet; (E) Opisthocystis goettei and (F) its copulatory organs. Ar, adenal rhabdoids; b, brain;
co, copulatory organ; cp, ciliated pits; dr, dermal rhabdoids; e, eye; ev, excretory vesicle; i, intestine; n, protonephridial duct; o, ovary; pg, pharyngeal
glands; ph, pharynx; pr, proboscis; s, stylet; t, testes; vg, granular vesicle; vs, seminal vesicle; y. yolk glands.
Chapter | 6 Flatworms: Turbellarians and Nemertea
ratio and is not very efficient. Consequently, turbellarians
may have a limited ability to adapt to low-oxygen condi-
tions. This conclusion appears to be true for most turbellar-
ians, and this limitation is often exploited in extraction of
specimens from samples[87,102]. However, the ability to cope
with reduced oxygen levels does not seem to differ sub-
stantially from that of other aquatic invertebrates. Oxygen
adaptability differs more among species adapted to diverse
Figure 6.9 A schematic representation of the main reproductive
organs and structures of freshwater Typhloplanoida. In some species,
additional structures may be present or the relative arrangement among the
organs may be slightly different. A, ascus; ad, atrium pouch; agi, atrium
genitale inferior; ags, atrium genitale superior; bc, bursa copulatrix; dc,
ductus copulatorius; de, ductus ejaculatorius; gp, gonopore; dc, ductus
communis; p, prostatic gland; rs, receptaculum seminis; o, ovary; t, testes;
ut, uterus; v, yolk gland; vg, vesicula granulorum; vs, vesicula seminalis.
Figure 6.10 Some representatives of Typhloplanoida: (A) Strongylostoma simplex; (B) copulatory organ with testes of S. simplex; (C) Prorhynchella
minuta; (D) a juvenile of Rhynchomesostoma rostratum; (E) Limnoruanis romanae; (F) sclerotized part of the copulatory organ, partly reverted (upper
picture), and the copulatory organ with the gonopore and glands (lower picture) in L. romanae (C modified from Ruebush[111]). Ar, adenal rhabdoids; e,
eye; g, glands; i, intestine; n, protonephridial duct; o, ovary; pg, pharyngeal glands; ph, pharynx; s, stylet; sph, sphincter; t, testes; vg, granular vesicle;
vs, seminal vesicle; y, yolk glands.
149
habitats than among major taxa (cf.[38]). For example, spe-
cies of Macrostomum normally found in stream headwa-
ters are more sensitive to low-oxygen levels, and perhaps
to high temperature, than are representatives of the same
genus from physically more fluctuating habitats (J. Kolasa,
personal observation). Some species, particularly in the
genus Phaenocora, have developed adaptations to cope
with anaerobic conditions. The presence of symbiotic algae
in Phaenocora typhlops, for instance, enhances its ability to
survive low-oxygen conditions[106]. This same species, as
well as a related P. unipunctata, produce hemoglobin, allow-
ing them to store oxygen during burrowing in the anoxic
mud. Many marine species, however, live in low-oxygen
sediments but have neither hemoglobin nor symbiotic algae.
Heitkamp[37,38] studied respiration rates of Opistomum
pallidum, two species of Mesostoma, and Dalyellia
viridis. He showed respiration to be related to body size
and shape (surface in a very flat Mesostoma ehrenbergi, or
weight in cylindrical Mesostoma lingua and other species).
Interestingly, Dalyellia viridis, which hosts symbiotic
zoochlorellae, had the highest respiration rate of all studied
species, possibly because their symbiotic algae contribute
to the observed rate. Respiration rate of microturbellarians
varied with temperature and diurnal rhythm[38].
Nutrition and digestion processes vary among turbel-
larians[44]. Lipid-storage capacity of Dugesia polychroa is
important to its population dynamics and competition, and
populations are regulated by intraspecific competition for
food, which affects individual size, cocoon production, and
cocoon viability[8]. Many microturbellarians appear to sup-
plement their energetic needs by living in endosymbiosis
with algae. Zoochlorellae occur in Phaenocora, Dalyellia,
150 Ecology and Classification of North American Freshwater Invertebrates

data). With only one exception, these species are not known
to produce resistant disseminules such as eggs or cocoons.

IV. Ecology of turbellarians

A. Life History
Most turbellarians develop directly; miniature worms
resembling the adult hatch from eggs attached to firm
substrates. These juveniles differ from adults chiefly in
Figure 6.11 Representatives of the Dalyellioida (Castrella) and the absence of reproductive organs. Some freshwater
Typhloplanida (others): (A) Castrella pinguis; (B) Phaenocora sp. (from
Churchill, Manitoba); (C) Mesostoma vernale (the habitus of this species is
turbellarians, however, are ovoviviparous (Mesostoma)
also characteristic of various Bothromestoma spp.); (D) Castrada hofmanni; or have a larval stage distinctly different from the adult
(E) Krumbachia hiemalis; (F) copulatory complex of Krumbachia ­hiemalis. (Rhynchoscolex). Numerous modifications of the life cycle
Ar, adenal rhabdoids; bc, copulatory bursa; bs, blind sac of the atrium; co, have evolved among turbellarians. Many seasonally occur-
copulatory organ; de, ductus ejaculatorius; gp, gonopore; i, intestine; m, ring species are univoltine, particularly those associated
matrix; n, protonephridial duct; o, ovary; ov, egg capsule; ph, pharynx; r,
seminal receptacle; s, stylet; t, testes; vs, seminal vesicle; y, yolk glands.
with temporary habitats or at the extremes of their geo-
graphical ranges. Other species are multivoltine, with the
number of generations depending on habitat availability
(see Heitkamp[39] for an excellent account).
Typhloplana, Castrada, and some other species. The nature
The diversity of life cycles observed in triclads remains
of the symbiotic association between algae and turbellar-
to be explained. Calow and Read[10] studied European
ians has been studied in a few species[77,92]. These indicate
representatives of the three families that are also repre-
that algae release substantial quantities of photosynthate
sented in North America: Dugesiidae, Planariidae, and
for the host turbellarian and that the host controls access to
Dendrocoelidae. They found that species breeding over
algal nutrients. This interdependence suggests an important
several seasons (Dugesiidae, Planariidae) invest less in
ecological constraint on the interaction of the symbionts.
reproduction than species that reproduce once and die
While triclads clearly avoid light, microturbellarians
(Dendrocoelidae).
appear to be either indifferent to it or positively phototac-
With rare exceptions, turbellarians are hermaphroditic.
tic (e.g., Typhloplana viridata which contains symbiotic
Various modifications of both egg formation and devel-
algae). However, if a sample is suddenly illuminated, many
opment are found and may include self-fertilization, as
microturbellarians will attempt to hide in sediments.
in Mesostoma ehrenbergi[25,32], protandry or progyny in
Desiccation of eggs can be tolerated by some spe-
many other species, or loss of either the male or female
cies, and it may be an obligatory factor in releasing them
gonad as in Catenulida[80]. All of these modifications have
from diapause and stimulating embryonic development.
implications for habitat adaptability, clonal diversity, rate
Similarly, some triclads such as Phagocata undergo
of population growth, and ability to colonize. Asexual
encystment as entire or fragmented animals. Temperature
reproduction by means of paratomy—that is, differentia-
may also have an important effect on reproduction. Clutch
tion of zooids that then separate by transverse division of
size, total offspring, and daily offspring all depend on tem-
the body—is common in several genera of microturbellar-
perature rather than food in Mesostoma lingua[21].
ians, particularly in Catenulida and Macrostomida. In mac-
Although the majority of turbellarians appear to have
roturbellarians, architomy (transversal fission of the caudal
some preferences for habitat, many species have adaptations
region which then differentiates a new individual after
conferring considerable flexibility within a limited range of
separation) occurs frequently in Dugesiidae and in some
physical conditions. Some members of the Catenulida are
species of Planariidae[10,80]. Asexual reproduction may be
found in habitats as different as fast-flowing streams and
the only mode of reproduction in some species. For exam-
temporary pools (e.g., Stenostomum leucops, Catenula
ple, sexual organs have never been observed in species of
­lemnae). Mesostoma species have been reported from warm
Stenostomum and in some populations of the common tri-
rice fields in the Sacramento Valley in California as well as
clad Dugesia tigrina.
from snow-melt pools at elevations above 2400 m[18].
Even though microturbellarians are delicate and fragile
organisms, they appear to have a significant potential for
dispersal. They can colonize artificial ponds situated on the
B. Distribution
roof of a building, for instance (Jan Ciborowski, personal While turbellarians practically by definition stand dis-
communication), within several weeks, half the ponds were tinct from the major parasitic platyhelminth taxa in being
colonized by six species of microturbellarians (unpublished free-living, a few European freshwater species such as
Chapter | 6 Flatworms: Turbellarians and Nemertea 151

ST

PH
M
I
B T

II M C

A N

CD
S
Y

ED B
O

D BC E
Figure 6.12 Microphotographs of live turbellarians and nemerteans: (A) Several partly framed individuals of Catenula sp. showing chains of
zooids; (B) frontal portion of body of Rhynchoscolex sp.; (C) Strongylostoma sp., a representative of Typhloplanoida; (D) squash preparation of the
portions of the reproductive system of Mesostoma craci, with the copulatory organ in the middle; (E) a frontal portion of body of a nemertean Prostoma
sp. B, brain; bc, bursa copulatrix; cd, common duct; ed, ejaculatory duct; prostatic granulations are visible in its proximity; i, intestine; m, mouth; n,
protonephridium; o, ovary; ph, pharynx; r, rhynchocoel; s, sensory pit; st, statocyst; t, testicle; y, yolk glands; I, first zooid; II, second zooid.

Varsoviella kozminski and Phaenocora beauchampi are
ectoparasitic on crustaceans. Several other freshwa-
ter forms, including triclads, occurring in Europe and
Australia are commensal on crustaceans and turtles[45].
The great majority of freshwater turbellarians, though, are
free-living and occupy the full variety of aquatic systems
including ponds, lakes, streams, hyporheic water, ditches,
and temporary puddles; in addition, some may be found in
semiterrestrial habitats such as water films among fallen
leaves in a mesic forest or in capillary water of soils in a
grassy meadow[84].
In North America, freshwater microturbellarians are
known as far north as Igloolik, a small island at 68°N
l­atitude off Melville Peninsula. Two species, one Mesostoma
and one unidentified mesostomid, were collected by P.D.N.
Hebert and associates there (unpublished). As many as six
other species are known from Alaska and in the area of
Hudson Bay in northern Canada[40,89]. Farther south, in tem-
perate climatic zones, the species diversity increases and
can easily reach 20 to perhaps 60 species in a single lake
or pond. For example, there are 23 species of flatworms
in the oligotrophic Mirror Lakes, New Hampshire[93],
and as many as 57 species occur in the polytrophic Lake
Zbechy in Poland[53]. Streams also have rich flatworm
fauna. In a short section of Wappinger Creek, an eastern
tributary of the Hudson River in New York, 15 species
152 Ecology and Classification of North American Freshwater Invertebrates
fo
e
b water microturbellarians, for example Bothrioplana sem-
i
g Most triclads from North America are described from
ov
s
Figure 6.13 A semischematic view of Prostoma sp. B, brain; e, eye;
fo, frontal organ; g, gonad; i, intestine; ov, egg; s, stylet.
were found[56]. A survey of several streams, including
springs and underground water in the same area revealed
32 species of microturbellarians, some of which are still
undescribed. Because of a dearth of systematic studies, few
microturbellarians are known from the West Coast, but at
least Mesostoma “lingua” and some other common species
are reported from rice fields in California[12,18].
Many species, particularly among the Catenulida and
the Macrostomida, are widely distributed worldwide,
while species of the “higher” turbellarians such as the
Typhloplanida and Tricladida are geographically more
restricted. About 3% of all species known from Europe are
designated as cosmopolitan in distribution[57], but studies
in the tropics and Southern Hemisphere[68] suggest this
figure is too low. As a whole, the triclad family Dugesiidae
occurs throughout the world, while the other major fresh-
water triclad families, the Planariidae and Dendrocoelidae,
have a Holarctic distribution[113].
Caves and underground waters hold rare and endemic
species, particularly of triclads[49]. Some troglobiotic tri-
clads and microturbellarians have recent or remote marine
origins[46,52]. An extensive study of European underground
triclads[33] revealed a suite of distinct distributional, repro-
ductive, and physiological characteristics. These often
include specialized underground microhabitat selection,
greater frequency of sexual reproduction, and slower metab-
olism as compared to epigean (surface-dwelling) triclads.
In contrast, a number of interstitial and underground
peri, Limnoruanis romanae, and Stenostomum pegephilum,
occur on more than one continent. Distributions of most
genera appear to be worldwide among microturbellarians,
but not triclads, whose many genera are known exclusively
from either North America, Eurasia, South America, or
Australia[2].
the eastern and southern United States and Canada[48].
Four triclad species inhabit Alaskan waters and 10 live
in Tennessee[14]. Several species of triclads with recent
marine origins occur in freshwater caves of Mexico[5,47].
Boreo-alpine distribution of many cold water spe-
cies has been tied to glaciations in Europe, and in North
America similar patterns also seem to occur[34]. At lower
latitudes, similarities in composition of microturbellar-
ian fauna between South American and North American
(cf.[63]) suggest considerable interchanges of species.
The ecological distribution of both microturbellarians
and triclads has been studied more intensively in Europe,
but most of the results are directly relevant to North
American fauna. The distribution of microturbellarians in
lakes has been investigated repeatedly[15,51,53,82,86,93,102,103].
These studies show that microturbellarians differenti-
ate among substrate types and depth zones. The flatworm
assemblages of sand and gravel bottoms are distinctly dif-
ferent from assemblages in habitats characterized by bot-
toms of silt, coarse organic matter, or vegetation. Plant
successional zones also differ in their species assem-
blages. Although the littoral zone has the greatest density
and diversity of flatworms, there is apparently no limit to
how deep some species may be found as long as oxygen
is available. For example, Otomesostoma auditivum lives
at a depth of 15–45 m in Echo Lake in the Sierra Nevada
range, but it occurs at a depth of 145 m in some European
alpine lakes[43].
Ecological differentiation of microturbellarians in run-
ning waters is more pronounced than in lakes. European
studies showed distinct assemblages of species associated
with springs, headwaters, and lower stretches of streams
and rivers (epirhithron, metarhithron, hyporhithron, and
potamon)[54,87]. These regularities in zonation of flatworm
assemblages are consistent with patterns observed among
other lotic taxa. These patterns, however, still need to be
related to ecological approaches to running waters, such
as the river continuum concept[99]. Among those classi-
cal zones, epirhithron shows the greatest species richness.
No comparable studies have been published on North
American microturbellarians. Zonation of triclads in
streams has been studied in greater detail, including exper-
imentation[78]. Competition, as well as thermal require-
ments were implicated as important factors separating
Chapter | 6 Flatworms: Turbellarians and Nemertea
several species of flatworms along a water course and in
standing waters[17,75,78].
Both microturbellarians and triclads live in habitats
where the physical conditions require additional adaptations
in comparison to an average pond or stream environment.
Wet mosses and leaves on the water edge, pooled water
at the base of bromeliad leaves, the high Arctic, ice-melt
ponds at elevations over 4000 m in central Asia, and hot
springs are each habitats for one or more species of flat-
worms. Representatives of freshwater families are also
commonly found in humid terrestrial habitats.
At a microhabitat scale, it appears that the number of
planarians is locally determined by the availability of shel-
ter. Brindle et al.[9] found that the number of Phagocata
gracilis was positively correlated with the size of stones
under which they rest during the day. This has implica-
tions at broader scales, whenever there is a systematic shift
in frequency or quality of shelter habitat due to geology
or history of perturbation. Understanding the geographi-
cal and ecological distribution of turbellarians is important
because their distribution has evolutionary and ecological
implications. Yet this understanding will suffer seriously
unless the dispersal capabilities of flatworms are assessed
in terms of distance and rates. Hebert and Payne[36] found
low levels of gene flow in Arctic Mesostoma. Their calcu-
lated dispersal rates are inconsistent with both the unpub-
lished data from Ciborowski’s experiment mentioned
earlier (Section III.B) and with common observations that
even newly created water bodies have a rich complement
of species. A series of questions may be posed. Is dispersal
different at different latitudes? Do dispersal strategies and
efficiency differ along major taxonomic lines? Are habitat
generalists better dispersers than specialists? Are species
associated with a particular habitat type (e.g., temporary
ponds) better dispersers compared to cave dwellers? If yes,
what in their life cycle makes them so? These questions
point out the limits in factual knowledge and ecological
interpretation of the many aspects of turbellarian dispersal.
C. Behavioral Ecology
Laboratory studies have shown that turbellarians have a
limited ability to learn simple tasks such as choosing white
over dark branches of a maze[66]. Observations of prey han-
dling (personal observation), however, indicate that their
learning ability may be much greater for tasks relevant to
natural conditions, such as prey or predator recognition.
For example, Dugesia dorotocephala (a planarian) recog-
nizes chemical cues generated by conspecifics injured by
fish and can associate those cues with pure fish cues[100]. It
appears they can learn to make a simple risk association.
Possible behavior modification or behavior determi-
nation associated with symbiosis is a potentially reward-
ing research area. Typhloplana viridata, a common spring
153
pond species, is unique in being positively phototactic.
This behavior may be associated with the presence of sym-
biotic algae, but just how it is controlled remains unknown.
Planarians (Tricladida) commonly rest underneath
stones as long as there is some space between the stone
and the underlying substrate. They may be active during
the day, but most remain hidden until it is dark.
D. Foraging Relationships, Predators,
and Parasites
Microturbellarians eat bacteria, algae, protozoa, and small
invertebrates, while triclads feed predominantly on larger
invertebrates. Scavenging is common in both morphologi-
cal groups. Various Mesostoma species have been studied in
detail and illustrate the diversity of feeding habits and tech-
niques within one genus. Leaf-shaped M. ehrenbergi usually
suspend themselves in the water column on a mucous fila-
ment attached to the surface film[32] and await a cladoceran
or an insect larva to come in contact. Using its front end,
M. ehrenbergi traps the prey with sticky mucus, wraps
around it, and proceeds to feed. M. lingua uses neurotoxins
to immobilize its prey in addition to mucus[21]. Mesostoma
can also glide on the surface of underwater objects or swim
directly in the water column. Both activities enhance its
chances of encountering prey. Unlike M. ehrenbergi, many
other species are spindle-shaped (e.g., M. arctica). These
species hunt more often on the bottom or among submerged
objects. Although several Mesostoma species studied spend
more than 50% of their time resting on various objects, they
can catch prey as soon as it comes close enough to create
vibrations in the water (J. Kolasa, personal observation).
At that point, the animal can erect the front portion of its
body and actively sweep it around until it contacts the prey.
Several species of Mesostoma, such as M. vernale, as well
as species of Bothromesostoma have a flat ventral surface
with which they cling to the surface film. These species are
more agile and much faster than other species of Mesostoma
and they prefer hunting at the water surface.
Immobilizing the prey and sucking its body flu-
ids is a common feeding behavior in microturbellarians
equipped with a strong pharynx rosulatus, or in triclads
with a pharynx plicatus. Many other turbellarians, how-
ever, swallow the victim whole or bite chunks from it.
For example, species of Stenostomum and Macrostomum
feeding on algae, protozoa, rotifers, and other meiofauna
usually swallow the prey whole. Some Macrostomum
species may be efficient predators on juvenile fresh-
water mussels. For example, Macrostomum tuba con-
sumed about one small mussel every 4 hours under
laboratory conditions; an alternative prey, the cladoceran
Ceriodaphnia reticulata, was eaten at a higher rate than
mussels when only one prey type was present, but at a
similar rate when both were present[20]. Others, such as
154 Ecology and Classification of North American Freshwater Invertebrates
Stenostomum predatorium and various Phaenocora spe-
cies may attack and partly consume larger turbellar-
ians[50], oligochaetes, or other soft-bodied invertebrates.
A few microturbellarians (e.g., Gyratrix hermaphroditus
and Prorhynchus stagnalis) use their copulatory stylets to
stab prey. The role of rhabdoids in capturing and subdu-
ing prey is, however, unclear. Wrona’s study[101] demon-
strated a lack of support for hypotheses that rhabdoids play
a major role in contacts with prey or predators in triclads.
In many microturbellarians, however, rhabdoids, and ade-
nal products in particular are channeled toward the anterior
of the body. The front end usually plays a crucial role in
exploration of the environment and in initiation of attack
against prey. This special position is highly suggestive of
more active applications of rhabdoids than the mere pro-
duction of mucus. In some turbellarians, the connection
between such glands and prey-capture proboscises shows
how these may play a role in prey capture[81]. Therefore,
the role of rhabdoids warrants further research. Rhabdoids
in certain marine acoels are known to be highly toxic and
to play a significant role in defense against predators[90].
Chemical detection of prey undoubtedly plays an
important role in feeding. Injured invertebrates attract spe-
cies of both triclads and microturbellarians under labora-
tory conditions. The presence of injured or recently killed
organisms allows scavenging by many individuals that
played no role in immobilizing the prey.
Turbellarians are subject to predation and parasitism.
Ciliates and flagellates are frequent parasites of Catenulida
and Typhloplanida, and nematodes have been found in
Lecithoepitheliata (Prorhynchus; J. Kolasa, unpublished).
Fish occasionally eat triclads[19,100]. Microturbellarians
(Phaenocora typhlops) may occasionally be eaten by other
invertebrates such as the chironomid Anatopynia[104] or
other turbellarians (e.g., Stenostomum predatorium).
The threat of predation by planarians appears to be seri-
ous enough to affect the developmental rate of salamander
eggs. In their study of predation by Phagocotus gracilis on
two Ambystoma species, Sih and Moore[91] found that the
flatworms, as well as chemicals they release, induce sala-
mander eggs to delay their hatching to a later time and thus
a larger size—a strategy interpreted as a predator defense.
E. Population Regulation and Density
It is not clear whether any single mode of population regu-
lation that applies to flatworms. Asexual reproduction of
Dugesia tahitiensis was density-dependent under laboratory
conditions[76]. The few studies conducted in Britain indicate
that intraspecific competition for food may be very impor-
tant for Phaenocora typhlops[105], while interspecific com-
petition may play a role in regulating several planarians[78].
In European Polycelis tenuis and perhaps in American Cura
foremanii (Girard), this regulation is expressed by a nega-
tive relationship between population growth and density. In
fact, long-term changes in the population size of Polycelis
suggest that limit cycles or cyclical fluctuations of the den-
sity due to intraspecific competition are involved[78].
Densities of microturbellarians vary according to the
habitat, season, and species. No major generalizations have
been made, but some patterns are beginning to emerge,
particularly along seasonal and successional axes. In
small, second- or third-order streams, the greatest densities
of microturbellarians are found in the lower reaches, that
is, in the metarhithron and hyporhithron, while the highest
richness is found in the epirhithron zone[54]. For example,
a mean density of 1280 individuals/m2 (of all species) was
recorded in Wappinger Creek, New York, in artificial sub-
strate after a week of colonization.
Density estimates of turbellarian populations are quite
variable. Schwank[87] found 80 microturbellarians per
liter in submontane streams in Germany, which translates
to over 4000 individuals/m, if one assumes an average
sample depth into the substrate of 5 cm. His data suggest
a corresponding density of triclads of 32 individuals per
liter and 1600/m2. In oligotrophic Mirror Lake, turbel-
larian densities varied from 40,000/m2 in shallow littoral
regions to almost zero in profundal zones; the lakewide
mean was 27,000/m2[93]. Other studies reported densities
of 800/m2 in Lake Michigan[70], 3500/m2 (annual mean) in
shallow littoral areas of Zbechy Lake in Poland[53], 3100/
m2 in Lake Paajarvi in Finland[41], to 9500/m2 in ponds
in Germany[39]. These values have to be interpreted cau-
tiously, however, in the light of seasonal succession and
abundance changes, which ordinarily produce the greatest
richness and abundance in early summer[3,16,93].
Planarians, for example Dugesia tigrina, as well as some
microturbellarians such as Mesostoma, engage in group for-
aging. In Dugesia, this strategy increases per capita ingestion
rates and leads to greater fecundity, although the benefits
peak at about four individuals and decline at higher flatworm
density[13]. Indeed, population dynamics, size structure,
asexual reproduction rates, and seasonal density fluctu-
ate in response to a variety of factors of which food supply,
intraspecific competition, and temperature appear to be most
important in this species[27] and illustrate the range of eco-
logical factors that may affect the great majority of species.
F. Functional Role in the Ecosystem
The functional role of species is typically discussed in
terms of their trophic interactions. All triclads are preda-
tory, as are most microturbellarians. Several inverte-
brates and vertebrates may consume triclads and are
a significant factor in their mortality. Mesostoma and
Rhynchomesostoma were subject to intense predation by
Dalyellia in temporary pools[39], but it is not clear how fre-
quent this predation is nor whether this predation results in
population regulation. However, triclads constitute a rela-
tively minor component of the diet of their predators[19].
Chapter | 6 Flatworms: Turbellarians and Nemertea
Triclads themselves consume isopods, midges, oligochae-
tes, caddisflies, mayflies, ostracodes, and cladocerans. The
ability to exploit various resources differs among species
and may result in strong competition with a simultaneous
expansion of invading species[26].
High densities of both triclads and microturbellarians
suggest that their role in biotic interactions of benthic com-
munities may be greater than their contribution to the diet of
other organisms. Densities of Mesostoma species (lingua)
in excess of 1000 individuals/m2 were observed in more
than 30% of the rice fields in a California study[18]. In some
cases, microturbellarians may regulate population dynam-
ics of zooplankton in ponds; for example, each Mesostoma
ehrenbergi consumed about two zooplankton individu-
als per day in one study[62] (estimated from Fig. 6.1). In
another study, this species consumed about 10 cladocerans
per day[88]. Mesostoma lingua might have major numerical
impact on mosquito larvae, cladocerans, and ostracodes in
rice fields but little effect on copepods[6,7]. More impor-
tant, perhaps, is the functional role of microturbellarians
as consumers of protozoa, rotifers, and algae, especially by
usually abundant Stenostomum leucops and similar forms.
Unfortunately, no quantitative data on this subject are avail-
able. Macrostomum predation on freshwater larval mussels
may reach pest levels in aquaculture (cf.[109]).
Although there are no explicit studies of the energy
flow through lake meiobenthos, the detailed study of the
energy budget of Mirror Lake[94] indicated this flow to be
equivalent to that of zooplankton. Furthermore, turbel-
larians, and Rhynchoscolex in particular, were a substan-
tial component of meiobenthos in Mirror Lake[93]. This
statement might be somewhat misleading because meiob-
enthos itself is a minor component of the whole budget.
A turbellarian role in the dynamics of standing and run-
ning waters is additionally hinted at when their diversity
and community structure are discussed. These change in
response to physical conditions, other species, and season.
Comparative research on the density and richness of tur-
bellarian assemblages in various water types might suggest
which factors have relatively greater influence in struc-
turing flatworm communities. Quantification and stand-
ardization of sampling procedures may be a major hurdle
for comparative ecologists. Different density estimates
obtained in quantification of lake turbellarians may be due
to seemingly minor differences in techniques[93].
V. Collecting, rearing, and
identification techniques for
turbellarians
A variety of techniques can be used to obtain qualitative
and quantitative samples of turbellarians: scooping with a
plankton net over aquatic vegetation or sediments; collecting
bottom sediments by means of bottom samplers such as
155
Ekman, Ponar, or multiple corers[87,93]; filtering water taken
from wells, springs, hyporheic interstitial, or ponds or pud-
dles using a plankton net[56]; or attracting turbellarians
(e.g., Mesostoma, triclads) into baited traps using pieces of
liver or injured aquatic insects[12,48]. In streams, a Surber
sampler or similar devices can be applied.
Samples should be cooled, topped with water, and
transported to the lab as quickly as possible. In the lab,
samples can be transferred to jars or beakers and left for a
couple of hours for the water to stagnate. When the water
clears, animals can be seen swimming toward the surface
or gliding on the glass where they can be picked up with
a pipette for further examination. Alternatively, turbellar-
ians can be directly picked from the sediments under a ster-
eomicroscope, but this procedure is more time-consuming.
Turbellarians can also be removed from mineral substrates
having a low organic matter content by gently heating the
sample on a hot plate so that the surface of the sediment
reaches 30–32°C[54]. Other methods of extracting flatworms
from sediments were developed for marine sands[65,71].
These methods are based on various combinations of anes-
thetization (with MgCl or ethanol), stirring water to sepa-
rate mineral particles, and sieving to catch worms. Some of
these methods work also on freshwater turbellarians.
Many triclads and microturbellarians can be main-
tained in the lab in small glass containers as long as the
water is regularly changed and appropriate food is pro-
vided. Synthetic pond water is best for common species.
Temperatures between 17 and 25°C may be adequate for
most species except for cold-water forms. Food require-
ments vary with the species, and a variety of easily available
aquatic invertebrates must be tested to ensure success. These
may include cladocerans, mosquito larvae (for Mesostoma,
Dugesia), oligochaetes (for Phaenocora), protozoa, or other
small turbellarians (Stenostomum)[55,108]. Often, pieces of
larger invertebrates such as Asellus, Gammarus, and Tubifex
can be successfully used. McConnell[66] provided many
useful suggestions for rearing triclads.
Most turbellarians can be identified by squash mounts
of live animals. Identification is difficult for a novice.
Squash mounts are prepared by placing a live individual in
a drop of water under a microscope coverslip. Next, excess
water is removed with a fine pipette or a strip of filter paper
until the specimen is immobilized by pressure of the cov-
erslip. That squeezing pressure can be varied by removing
or adding water. Such mounts, with the appropriate degree
of squeezing, reveal the arrangement and appearance of
internal organs—information usually necessary for species
determination. Whenever further study is desirable, ani-
mals can be fixed and preserved as in the following steps.
1. Anesthetizing with 7% ethanol, 0.1% chloretone, 1%
hydroxylamine hydrochloride, or slowing their loco-
motion by placing them in a small volume of water on
ice (this step is optional);
156 Ecology and Classification of North American Freshwater Invertebrates

2. Killing with hot fixatives such as Stieve’s, Gilson’s,
Bouin’s, room temperature 70% ethanol, or (optional)
killing cooled worms with glutaraldehyde (see electron
microscopic techniques for details);
3. Rinsing mercury residues of the above fixatives
(except Bouin’s) with 50% ethanolic solution of
iodine; and
4. Storing specimens in 70% ethanol. Animals so prepared
can be sectioned and analyzed with standard histologi-
cal methods, including staining with Mallory’s stain,
Delafield’s hematoxylin, and other common stains.
VI. Identification of North
American Genera of turbellarians
The following key uses characters that can be seen in whole
mounts of living animals. Some that distinguish major
systematic groups may be less easily discerned by inex-
perienced observers, but they can be applied with greater
confidence because of their systematic ­ significance. An
identification reached with this key should be ­ confirmed
by comparing the specimen at hand with the original
description of the species or with monographic reviews
from the taxonomic literature. Monographs by Luther[59–
61]
(for Dalyelliidae, Macrostomidae, Typhloplanidae),
Nuttycombe[72], Nuttycombe and Waters[73] (for Catenula,
Stenostomum), Ferguson[24] (for Macrostomum), and
Gilbert[31] (for Phaenocora) are ­particularly useful.
A. Taxonomic Key to Orders and Suborders
of Turbellarians
As some characters may be difficult to determine, it may
be advisable to match the general body plan of the individ-
ual being identified with one of the pictures in Fig. 6.1a–h.
For additional schematics, consult Cannon’s[11] world
guide to turbellarians.

1a. Simple female gonad; egg entolecithal (Acoela, Macrostomida, Catenulida; freshwater members of the latter two often reproduce asexu-
ally and ovary is absent) (Figs. 6.1a–c, 6.12a,b) . ...........................................................................................................................................2
1b. Heterocellular female gonad, with yolk-producing part separate from the oocyte-producing part (all other Turbellaria) (Figs. 6.1d–h, 6.2) 4
2a (1a). Mouth opens directly or through a pharynx simplex into the body; no protonephridia; no distinct intestine; gonads without clear walls
(Fig. 6.1a); rare in freshwater ............................................................................................................................................................... Acoela
[Only one species (unidentified) collected in North America (from Mirror Lake in New Hampshire[93].]
2b. Mouth opens to a pharynx simplex; protonephridia present; epithelial, ciliated intestine present . ...............................................................3
3a (2b). Protonephridia with a single, central excretory duct, often two or more zooids; male gonopore, if present, situated on the dorsal side and
in the front of the body; ovary without oviducts and supplementary organs; statocyst present in some species (Figs. 6.1b, 6.12a,b) . ..........
....................................................................................................................................................................................... Catenulida (key VI.B)
3b. Protonephridia with a pair of main excretory ducts; male gonopore on the ventral side of the posterior part of body; female gonopore
usually separate and in front of the male one, more than one zooid in the family Microstomidae only; no statocyst (Figs. 6.1c, 6.5a,b) . ....
................................................................................................................................................................................. Macrostomida (key VI.C)
4a (1b). Single germovitellarium where ova are surrounded by yolk cell epithelium; pharynx variabilis; copulatory organ armed with a sclero-
tized stylet near the pharynx (Figs. 6.1d, 6.2c) . ............................................................................................... Lecithoepitheliata (key VI.D)
4b. Ova not surrounded by yolk epithelium; ovary separate or combined with vitellarium (Figs. 6.1e–h, 6.2a,b) .............................................5
5a (4b). Pharynx plicatus or variabilis; testicles and ovaries dispersed; germ cells dispersed or aggregated (Fig. 6.1h)...............................................
....................................................................................................................................Proseriata, Prolecithophora, and Tricladida (key VI.E)
5b. Pharynx bulbosus (doliiformis or rosulatus, Fig. 6.1e–g) . .............................................................................................................................6
6a (5b). Pharynx doliiformis, directed forward (Fig. 6.1e) ..................................................................................................... Dalyellioida (key VI.H)
6b. Pharynx rosulatus directed ventrally or, exceptionally as in Phaenocora, ventrally and forward .................................................................7
7a (6b). Without a proboscis (Fig. 6.1f) ..............................................................................................................................Typhloplanoida (key VI.F)
7b. Proboscis present (Fig. 6.1g) .............................................................................................................................. Kalyptorhynchia (key VI.G)

B. Taxonomic Key to Catenulida

1a. Brain simple, compact, oval . ..........................................................................................................................................................................2
1b. Brain composed of paired frontal and posterior lobes; a group of sensory cells in front of the brain, often arranged pseudometamerically
in parallel rows Stenostomidae .......................................................................................................................................................................5
2a (1a). Brain situated usually at the base of but not further than in the middle of prostomium (frontal body section without the intestine); with
ciliated ventral or lateroventral furrow separating prostomium from the rest of body Catenulidae . .............................................................3
2b. Brain situated in the first one-third of prostomium Chordariidae ................................................................................................Chordarium
3a (2a). A ring of strongly ciliated longitudinal grooves developed on the prostomium (Fig. 6.3a) . ............................................... Catenula turgida
Chapter | 6 Flatworms: Turbellarians and Nemertea 157

3b. No such grooves present .................................................................................................................................................................................4

4a (3b). Clearly open intestine extends approximately, to the middle of the postpharyngeal section of body (Figs. 6.3b,c, 6.12a) ......Catenula (in part)
4b. Open intestine extends much further, leaving no more than one-fifth to one-fourth of the postpharyngeal section closed; usually the
intestinal lumen of individual zooids is connected ...................................................................................................................... Dasyhormus
5a (1b). Adult individuals composed of two or more zooids; gonads often absent .....................................................................................................6
5b. Adult individuals (with gonads) without signs of asexual divisions; with long prostomium, ciliated pits absent (Figs. 6.3d,f,
6.12b).........................................................................................................................................................................................Rhynchoscolex
6a (5). A section of the intestine near the pharynx with a strong ring of muscles (Fig. 6.4e) .........................................................Myostenostomum
6b. No such structure, intestine uniformly ciliated (Fig. 6.4a–d) ..................................................................................................... Stenostomum

C. Taxonomic Key to Macrostomida

1a. Ciliated pits present; intestine extends in front of mouth; numerous zooids separated by division planes (Fig. 6.5b,c) Microstomidae . ......
.....................................................................................................................................................................................................Microstomum
1b. No ciliated pits; intestine entirely behind the pharynx; no signs of asexual division (Fig. 6.5a,d,e) Macrostomidae ..............Macrostomum

D. Taxonomic Key to Lecithoepitheliata

1a. Stylet of the copulatory organ straight; no eyes . ......................................................................................................................... Prorhynchus
1b. Stylet of the copulatory organ curved; eyes present except in some cave forms . ................................................................. Geocentrophora

E. Taxonomic Key to Prolecithophora, Proseriata, and Tricladida

1a. Mouth situated in front and directed forward ................................................................................................................................Hydrolimax
1b. Mouth situated elsewhere and directed backward or ventrally . .....................................................................................................................2
2a (1b). Mouth situated near the middle and directed ventrally; statocysts present ..............................................................................Otomesostoma
2b. Mouth directed backward; no statocyst ..........................................................................................................................................................3
3a (2b). Intestine split into two branches behind the pharynx (Figs. 6.6, 6.7) Tricladida . ..........................................................................................4
4a (3a). Internal pharyngeal muscles in two distinct layers (Planariidae) (Fig. 6.6) ...................................................................................................5
4b. Internal pharyngeal muscles in one layer of mixed longitudinal and circular fibers (Fig. 6.6) Dendrocoelidae .........................................12
5a (4a). Oviducts, separate or united, open into end part of the bursa stalk ................................................................................................................6
5b. Oviducts united, the common oviduct opens in the genital atrium (Fig. 6.6) ................................................................................................7
6a (5a). Testes extend to the level of pharynx ....................................................................................................................................................... Cura
6b. Testes extend to the posterior end of body . ........................................................................................................................................ Dugesia
7a (5b). Numerous eyes arranged in a band around the anterior end of body (Fig. 6.7g) .............................................................................. Polycelis
7b. Eyes absent or one pair only; if numerous, then not along the head margin ..................................................................................................8
8a (7b). Adenodactyl present (Fig. 6.6) .......................................................................................................................................................... Planaria
8b. Adenodactyl absent . .......................................................................................................................................................................................9
9a (8b). Anterior end with an adhesive organ ............................................................................................................................................................10
9b. Anterior end without an adhesive organ .......................................................................................................................................................11
10a (9a). Body elongated, flat, with a well-developed postpharyngeal section . ....................................................................................... Sphalloplana
10b. Body turtle-shaped, with reduced postpharyngeal section ................................................................................................................... Kenkia
11a (9b). Prepharyngeal, fused testes in male phase; large atrium lined by papillose epithelium in female phase; head round and broad; the anterior
ramus of the intestine does not reach beyond eyes ......................................................................................................................... Hymanella
11b. Other set of characters .................................................................................................................................................................... Phagocata
12a (4b). Anterior end of the body with a deeply invaginated adhesive organ ........................................................................................... Macrocotyla
12b. Adhesive organ absent, or an adhesive disk present .....................................................................................................................................13
13a (12b). Penis papilla present .....................................................................................................................................................................................14
13b. Penis papilla absent, bulb large . .................................................................................................................................................Rectocephala
14a (13a). Penis bulb rounded, containing a seminal vesicle .................................................................................................................Dendrocoelopsis
14b. Penis bulb elongated, contains a prostatic vesicle ............................................................................................................................ Procotyla
158 Ecology and Classification of North American Freshwater Invertebrates

F. Taxonomic Key to Typhloplanoida

The basic plan of the freshwater typhloplanoid reproductive system is given in Fig. 6.9.
1a. Yolk glands paired, situated on both sides of the intestine .............................................................................................................................2
1b. Yolk glands developed as single strand of cells above the intestine (Fig. 6.10e,f) ...................................................................... Limnoruanis
2a (1a). Pharynx developed as a typical, round or slightly elongated pharynx rosulatus and oriented ventrally or somewhat forward .....................3
2b. Pharynx elongated or cylindrical, oriented backward; copulatory organ with a spiny cirrus (Fig. 6.5h) . .................................... Opistomum
3a (2a). Testes under yolk glands . ...............................................................................................................................................................................4
3b. Testes above yolk glands ..............................................................................................................................................................................11
4a (3a). Protonephridial ducts open separately on the body surface (Protoplanellinae) (as in Fig. 6.10c) . ................................................................5
4b. Protonephridial ducts combined with mouth or gonopore (as in Fig. 6.10a) .................................................................................................7
5a (4a). Front extremity with a central depression and set slightly apart by two lateral, ciliated pits; pharynx in the first one-third of body (Fig.
6.10c) .........................................................................................................................................................................................Prorhynchella
5b. Front extremity not set apart, rounded; pharynx in the second to third portion of body ................................................................................6
6a (5b). Reproductive complex with a bursa copulatrix; ductus ejaculatorius usually surrounded by gelatinous matrix (Fig. 6.11e,f; cf., Fig.
6.9) ................................................................................................................................................................................................Krumbachia
6b. Bursa copulatrix absent . ............................................................................................................................................................. Amphibolella
7a (4b). Protonephridial ducts joined and combined with mouth; no retractable proboscis Typhloplanidae ..............................................................8
7b. Protonephridial ducts open into the gonopore; retractable front extremity (Fig. 6.10d) ..................................................Rhynchomesostoma
8a (7a). With a bursa copulatrix and an atrium copulatorium (Fig. 6.11d) . ...................................................................................................Castrada
8b. One of the above organs missing ....................................................................................................................................................................9
9a (8b). With eyes (Figs. 6.10a, 6.12c) .................................................................................................................................................Strongylostoma
9b. No eyes .........................................................................................................................................................................................................10
10a (9b). Atrium copulatorium absent; bursa copulatrix present .............................................................................................................Typhloplanella
10b. Both bursa copulatrix and atrium copulatorium absent; always with green algae; pharynx in the first half of body; copulatory organ pear-
shaped ...........................................................................................................................................................................................Typhloplana
11a (3b). Pharynx developed as a typical, round pharynx rosulatus ............................................................................................................................13
11b. Pharynx barrel-shaped, directed clearly forward, and situated near the front extremity (Fig. 6.11b) ..........................................................12
12a (11b). Ductus copulatorius absent; inferior genital atrium inconspicuous (Fig. 6.11b) . ........................................................................ Phaenocora
12b. With ductus copulatorius; exceptionally large inferior atrium; occurs in sulfuric springs ................................................Pseudophaenocora
13a (11a). An elongated supplementary organ, the ascus, present in the proximity of the gonopore; or with a well-defined gland complex in its
place (Ascophorinae ) (cf., Fig. 6.9) .............................................................................................................................................................14
13b. Ascus absent .................................................................................................................................................................................................16
14a (13a). Pharynx in the middle of the first half of body ...............................................................................................................................Protoascus
14b. Pharynx approximately in the middle of body or slightly behind ................................................................................................................15
15a (14b). Gonopore near the middle of body ................................................................................................................................................. Ascophora
15b. Gonopore in the third posterior part of body ............................................................................................................................Dochmiotrema
16a(13b). Protonephridial ducts open separately on the body surface; eyes often present . .........................................................................Olisthanella
16b. Protonephridial ducts open into mouth .........................................................................................................................................................17
17a (16b). With a ventral pit and a canal connecting the bursa copulatrix to the ovovitellin duct; often flat on the ventral side and strongly convex
on the dorsal . .......................................................................................................................................................................Bothromesostoma
17b. Without the above characters (Figs. 6.11c, 6.8b, 6.12d) . .............................................................................................................. Mesostoma

G. Taxonomic Key to Kalyptorhynchia and Similar Forms

1a. Excretory vesicle absent; protonephridial ducts open directly at the surface of body; single ovary (Fig. 6.8c) ............................................2
1b. Protonephridial ducts open into an excretory vesicle in the rear end of body; paired ovary (Fig. 6.8e,f) . ................................ Opisthocystis
2a (1a). Copulatory organ with an almost straight stylet (Fig. 6.8c,d) ............................................................................................................Gyratrix
2b. Copulatory organ with a curved stylet ..........................................................................................................................Microcalyptorhynchus

H. Taxonomic Key to Dalyellioida

1a. With a paired ovary; copulatory organ with a straight, funnel-like tube ..........................................................................................Provortex
1b. With a single ovary; copulatory organ with a more of less complex sclerotized structure (e.g., Figs. 6.5f,g, 6.11a) ....................................2
Chapter | 6 Flatworms: Turbellarians and Nemertea 159

2a (1b). Sclerotized structure directly attached to the rest of the copulatory organ . ...................................................................................................3
2b. Sclerotized structure in a separate pocket adjacent to the bulb of the copulatory organ (Fig. 6.11a) ...............................................Castrella
3a (2a). Sclerotized structure typically with two handles and two lateral branches carrying spines, auxiliary parts may be present (Fig. 6.5f,g) 4
3b. Sclerotized structure different; usually in the form of a spiny ring ............................................................................................... Gieysztoria
4a (3a). One to several eggs in the parenchyme; normally, older individuals green due to symbiotic zoochlorellae ....................................Dalyellia
4b. Never more than one egg; no zoochlorellae . ............................................................................................................................ Microdalyellia

VII. Nemertea: general characteristics and external and internal

anatomical features
Nemerteans are unsegmented worms possessing an eversi-
ble muscular proboscis resting in the rhynchocoel, which is
a walled, longitudinal, dorsal coelomic cavity (Figs. 6.13,
6.14, see also Fig. 6.12e, and section VI.F). Unlike the tur-
bellarians, nemerteans have an anus and a coelomic circula-
tory system (identified as a blood-vascular system in older
literature before its coelomic nature was discerned through
electron microscopy)[97]. A monolayered, ciliated epider-
mis and several layers of muscles constitute the body wall.
The excretory system is protonephridial. Sensory structures
include eyes, statocysts (absent in freshwater nemerteans),
and cerebral, frontal, and lateral neural organs.
The reproductive system is much simpler than in turbel-
larians. Freshwater nemerteans are hermaphroditic and often
protandric. Spermatozoa and ova are produced in numerous
gonads situated between diverticulae of the intestine. As a
rule, fertilization is external, and self-fertilization can occur.
Development is direct. Most information provided in this sec-
tion on nemerteans is based on the review of freshwater and
terrestrial nemerteans by Moore and Gibson[69]. That review is
an invaluable source of data and references for anyone work-
ing on the ecology and biology of freshwater nemerteans.
Twenty-two known species of freshwater nemerteans[95] are
classified into two, possibly polyphyletic families and six gen-
era[69]. In the family Heteronemertea, three monotypic genera
are recognized: Planolineus Beauchamp 1928, Siolineus du
Bois-Reymond Marcus 1948, and Apatronemertes Wilfert
and Gibson 1974. In the family Hoplonemertea, the genus
Prostoma Duges 1828 contains most of the species. Two
other genera, Campbellonemertes Moore and Gibson 1972
and Potamonemertes Moore and Gibson 1973 are known
from islands in the southern Pacific, Campbell Island and
South Island, New Zealand, respectively.
VIII. Ecology of Nemertea
A. Life History and General Ecology
Most nemerteans are marine, with only a small number of
terrestrial or freshwater species. The freshwater forms are
benthic, predatory worms 10–40 mm in length and of pink
coloration. They use the proboscis, which produces sticky
mucus laced with neurotoxins, to capture prey. Prostoma
species feed readily on oligochaete worms and occasion-
ally on crustaceans, nematodes, turbellarians, midge lar-
vae, and other small invertebrates[67]. Feeding activity is
most intense at night. Little is known about habitat selec-
tion by freshwater nemerteans. North American Prostoma
species have been found in both streams and lakes, includ-
ing Lake Huron. In lakes, Prostoma species appear to be
associated with filamentous algae.
Even though some nemerteans have strong regenerative
capabilities, reproduction is exclusively sexual, with both
male and female organs present concurrently. Since only
limited quantitative, field studies have been conducted
on the biology of freshwater nemerteans, mechanisms of
population regulation remain unknown. Clearly, Prostoma
species can reproduce rapidly, with up to 210 eggs per
reproductive episode throughout the year, as long as the
temperature is above 10°C[107].
A study of one population of Prostoma in England sug-
gests that population density increases during the cold months
of the year and declines during the summer, with smaller
individuals recorded during the winter[107]. It is not clear,
however, if this pattern is general or due to trampling by cat-
tle of the shore vegetation where the study was conducted.
Distribution of nemerteans in North America is not well
known. Strayer[93] reported Prostoma rubrum from Mirror
Lakes, New Hampshire. However, P. rubrum is no longer
a valid species[28], and this observation has to be treated
as Prostoma sp. Prostoma canadiensis is known from one
site in Lake Huron[29], where it occurs to a depth of 20 m.
Prostoma species are probably much more common in
freshwaters than the infrequent records from eastern North
American might indicate. A Prostoma species (P. rubra) was
also reported in vegetation of slow coastal streams, marshes,
and estuarine pools in St. Vincent, Caribbean[35]. So far, no
Prostoma have been found in temporary water bodies.
Three established species of nemerteans are known from
North America (Fig. 6.14); a fourth, P. asensoriatum, is a
questionable species. Distribution of these species in North
America is summarized in Fig. 6.15, but they are ­ probably
present in most standing and running waters. With the excep-
tion of the dubious P. asensoriatum, all three remaining spe-
cies have been recorded in other parts of the world, including
160 Ecology and Classification of North American Freshwater Invertebrates

B. Physiological Adaptations
cg
Osmoregulation is an ecologically important function in
nemerteans, as in all freshwater invertebrates with perme-
able body walls. It is controlled by the cerebral organs and
involves several organs and enzymatic systems associated
with blood vessels[69]. Well-developed nephridia may play
a role in osmoregulation as well as in the removal of nitrog-
enous wastes (but the latter has not yet been demonstrated).
fo rd st rl oe Gases are exchanged across the surface of the body. The
pr bw role of the coelomic channels (“blood vessels”) in oxygen
rc
dc transport is unclear. Nemerteans, like turbellarians, release
cg
copious mucus whose various functions probably include
ie defense, locomotion, physiological barrier, and encystment.

fo
rd oe ep
vc st
Figure 6.14 Sections through the anterior extremity of three North
American nemerteans: (A) Prostoma eilhardi; (B) P. graecense; (C) P.
canadiensis (after Gibson and Moore[28,29]). Bw, body wall musculature;
cg, cephalic gland lobular region; dc, dorsal cerebral commissure; ep,
­epidermis; fo, frontal organ; id, improvised ducts from cephalic gland
lobules; oe, oesophagus; pr, proboscis; rc, rhynchocoel; rd, rhynchod-
aeum; rl, rhynchodaeal longitudinal muscle fibers; st, stomach; vc, ven-
tral cerebral commisure.
C. Behavioral Ecology
There are no studies devoted exclusively to the behavior of
freshwater nemerteans. Nevertheless, some observations
on feeding behavior, escape reactions, and locomotion are
available. Adult nemerteans can only crawl, while small
juveniles also swim. Forward locomotion of Prostoma
may involve ciliary movement alone, ciliary movement in
combination with muscular waves, sinusoidal curves when
the animal pushes through the vegetation, or peristalsis
when the animal moves backward. The proboscis provides
the fastest movement forward. This long organ is rapidly
everted and attached to substrate, and the body is then
pulled toward the point of attachment.
Typically, prey are captured by the sticky proboscis
and then swallowed whole through the mouth, which can
be widely distended[67]. The mechanism of prey detection
is poorly understood. Terrestrial nemerteans ambush their
prey, which they detect through mechanical stimuli.

D. Functional Role in the Ecosystem

Figure 6.15 Known distribution of Prostoma species in North
America (after Gibson[112]; modified and updated). C, P. canadiensis; E, P.
eilhardi; A, P. asensoriatum; G, P. graecense; U, unidentified Prostoma.
South America, Europe, Africa, and Australia. Zoologists
do not know whether the worldwide pattern of distribution
is natural or, instead, a result of recent human-mediated
dispersal. Analysis of genetic diversity of various popula-
tions should shed light on this question.
By necessity, the importance of nemerteans in ecosystems
can be inferred only indirectly from their trophic position
and relative densities. Densities of Prostoma species in
Wappinger Creek, a tributary of the Hudson River, ranged
between 50 and 590 individuals/m2, which constitutes a
small fraction of all predatory invertebrates identified at the
study site (J. Kolasa, personal observation). Similarly, low
densities of P. canadiensis (up to 140/m2) occurred on grav-
elly and muddy substrates in Lake Huron. In view of the
limited knowledge of nemertean ecology, almost any study
will provide useful information. In particular, observations
of phenology, population dynamics, and habitat selectivity
are needed to better evaluate the role of nemerteans in fresh-
water communities. Also needed are studies of reproduction
(e.g., ecological significance of protandry and resting eggs)
and cues triggering life-stage changes, the latter needed to
understand the evolutionary ecology of nemerteans.
Chapter | 6 Flatworms: Turbellarians and Nemertea 161

IX. Collection, culturing, and
preservation
Collection of nemerteans is similar to collection of other
soft-bodied benthic invertebrates such as turbellarians or
oligochaetes living in the substrate or on aquatic plants.
Placing samples in beakers or jars and topping with water
induces nemerteans to crawl toward the air–water interface
where they can be collected with a pipette. Sieving and
vigorous washing may damage specimens.
Freshwater nemerteans are relatively easy to culture.
They may be maintained in aquaria or even in petri dishes
in clean, oxygenated water and on a diet of live aquatic
invertebrates. Chopped tubificid oligochaetes provide a
good diet. The worms will also reproduce in such cultures.
Nemerteans can be preserved in 80% ethanol. Before
preservation, however, worms should be narcotized using
7% ethanol or chloretone until their movement ceases.
According to Moore and Gibson[69], gin can be used in
the field if pure ethanol is unavailable, but only as a last
resort!
X. Taxonomic Key to Species of
Freshwater Nemerteans
This key is an adaptation of keys by Gibson and
Moore[29,30] to freshwater nemerteans worldwide, modi-
fied specifically for identifying North American Prostoma.
In the event this key does not lead to a reasonable identifi-
cation, perhaps because a specimen is of a species not yet
recorded in North America, consult their original keys.

1a. Cephalic glands open via frontal organs and by improvised ducts (Fig. 6.14c); esophagus distinct but unciliated; rhynchodaeum with
isolated longitudinal muscle strands; proboscis with 12 nerves .................................................... Prostoma canadiensis Gibson and Moore
1b. Cephalic glands open only via frontal organ, no improvised ducts ................................................................................................................2
2a (1b). With a distinctive ciliated esophagus; cephalic glands reach back to the brain (Fig. 6.14b); rhynchodaeum with a well-developed layer of
longitudinal muscle . ....................................................................................................................................................P. graecense (Bohmig)
2b. Different combination of characters; indistinct and unciliated esophagus (Fig. 6.14a); rhynchodaeum without specifically associated
layer of longitudinal muscle fibers; proboscis with 9–10 nerves ...........................................................................P. eilhardi (Montgomery).

XI. Selected References
11. Cannon LRG. Turbellaria of the World. A Guide to Families and
Genera. Brisbane: Queensland Museum; 1986.
24. Ferguson FF. A monograph of the genus Macrostomum O. Schmidt
1848. Part II. Zool Anzeiger. 1939;126:131–144.
28. Gibson R, Moore J. Freshwater nemerteans. Zool J Linnean Soc.
1976;58:177–218.
42. Hyman LH. The Invertebrates: Platyhelminthes and Rhynchocoela,
the Acoelomate Bilateralia. New York: McGraw-Hill; 1951.
48. Kenk R. Freshwater planarians (Turbellaria) of North America. Pages
1-81 in: EPA Biota of Freshwater Ecosystems, Identification Manual
Vol. 1; 1972.
50. Kepner WA, Carter JS. Ten well-defined new species of
Stenostomum. Zool Anzeiger. 1931; 93:108–123.
56. Kolasa J, Strayer D, Bannon-O’Donnell E. Microturbellarians from
interstitial waters, streams, and springs in southeastern New York.
J North Am Benthol Soc. 1987;6:125–132.
57. Lanfranchi A, Papi F. Turbellaria (excl. Tricladida). In: Illies J, ed.
Limnofauna Europaea. Stuttgart: G. Fisher Verlag; 1978:5–15.
59. Luther A. Die Dalyelliiden. Acta Zool Fennica. 1955;87:1–337.
50. Luther A. Die Turbellarien Ostfennoskandiens. I. Acoela, Catenulida,
Macrostomida, Lecithoepitheliata, Prolecithophora, und Proseriata.
Fauna Fennica. 1960;7:1–155.
51. Luther A. Die Turbellarien Ostfennoskandiens IV. Neorhabdocoela 2.
Typhloplanoida: Typhloplanidae, Solenopharyngidae und Carcharo­
dopharyngidae. Fauna Fennica. 1963;16:1–163.
72. Nuttycombe JW. The Catenula of the Eastern United States. Am
Midi Nat. 1956;55:419–433.
73. Nuttycombe JW, Waters AJ. The American species of the genus
Stenostomum. Proc Am Phil Soc.. 1938;79:213–301.
81. Rieger RM, Tyler S, Smith JPS III, Rieger GE. Platyhelminthes:
Turbellaria. In: Harrison FW, Bogitsh BJ, eds Microscopic Anatomy
of Invertebrates. Vol. 3: Platyhelminthes and Nemertinea. New
York: Wiley-Liss; 1991:7–140.
87. Schwank P. Turbellarien, Oligochaeten and Archianneliden des
Breitenbachs und anderer oberhessischer Mittelgebirgsbäche. II.
Die Systematik und Autökologie der einzelnen Arten. Archiv für
Hydrobiol. 1981;62(Supplement):1–85.
98. Tyler S, Schilling S, Hooge M, Bush LF (comp.). Turbellarian taxo-
nomic database. Version 1.5; http://turbellaria.umaine.edu/; 2008.
113. Sluys R, Kawakatsu M. Towards a phylogenetic classification of
dendrocoelid freshwater planarians (Platyhelminthes): a morpholog-
ical and eclectic approach. J Zool Syst Evol Res. 2006;44:274–284.
Acknowledgments
We are very grateful to L. R. G. Cannon, J. Moore, R.
Gibson, A. P. Mead, R. Kent, S. Schwartz, and D. Strayer
for helpful comments on the manuscript, additional ref-
erences, and permission to reproduce figures, glossary
entries, and fragments of keys.