ORIGINAL ARTICLE /ARTÍCULO ORIGINAL

A NEW SPECIES OF TEMNOCEPHALA (PLATYHELMINTHES, TEMNOCEPHALIDA)

ECTOSYMBIONT ON NERITINA ZEBRA (MOLLUSCA, NERITIDAE)

FROM THE BRAZILIAN AMAZONIA

UMA NOVA ESPÉCIE DE TEMNOCEPHALA(PLATYHELMINTHES,

TEMNOCEPHALIDA) ECTOSIMBIONTE SOBRE NERITINA ZEBRA (MOLLUSCA,

NERITIDAE) DA AMAZÔNIA BRASILEIRA

1 2 3

Samantha A. Seixas , José F. R. Amato & Suzana B. Amato

1Setor de Evolução e Ecologia, Programa de Pós-Graduação em Ambiente e Desenvolvimento, Centro Universitário

UNIVATES, Rua Avelino Tallini 171, Prédio 8, salas:102/104, Bairro Universitário, CEP 95900-000, Lajeado, RS, Brasil.

PNPD/CAPES Post-Doctoral Fellow. E-mail: seixas.sa@gmail.com

2Rua Cabral, 875/301, Bairro Rio Branco, CEP 90420-121, Porto Alegre, RS, Brasil. E-mail: josefelipeamato@gmail.com

3Laboratório de Helmintologia, Departamento de Zoologia, Instituto de Biociências, Universidade Federal do Rio Grande do

Sul, Av. Bento Gonçalves 9500, Prédio 43435, sala 208, Bairro Agronomia, CEP 91501-970, Porto Alegre, RS, Brasil. E-

mail: sbamato@ufrgs.br

Neotropical Helminthology, 2015, 9(1), jan-jun: 41-53.

ABSTRACT

Keywords: mollusks - Neotropic Region - Taxonomy - temnocephalids.

Temnocephala sp. n., an ectosymbiont of the mollusk Neritina zebra (Bruguière,

1792), is described from the State of Pará, Brazilian Amazonia. Representatives of Neritidae

Lamarck, 1809, usually euryhaline, are adapted to habitats with a certain degree of salinity. Up to

now, all species of Temnocephala epibiont on mollusks were described/recorded on

Ampullariidae Gray, 1824, hosts strictly from freshwater, as all previous hosts of Temnocephala

spp. Twenty-one mollusks were collected from Rio Tocantins, Municipality of Cametá. The new

species differs from all the epibiont temnocephalan species of mollusks described so far, by

having the following characters: 1. small body size; 2. cirrus simple and short, different from the

cirrus of other epibiont species of mollusks that, unless specific variations, has a 'brush' format,

where the boundary between the shaft and the introvert is well marked and the introvert has a

swelling portion; 3. feminine reproductive system simple and inconspicuous; 4. vagina not

muscular, without differentiation between the proximal and distal portions; 5. vesicula

intermedia small, replacing the seminal receptacles; and 6. vaginal sphincter single and

symmetric. The eggs of the temnocephalans were not found, inside or outside the shells and on

the opercula. This is the first record of a species of Temnocephala epibiont on Neritid mollusks

and the first record of an euryhaline host for any Temnocephalida.

euryhalina

ISSN Versión impresa 2218-6425 ISSN Versión Electrónica 1995-1043

INTRODUCTION

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015

Up to the present, all known mollusk hosts of

species of Temnocephala Blanchard, 1849 are

ampullariid gastropods. This family has a large

number of species distributed in five genera,

and it is quite common in freshwaters of the

Neotropic Region (Cowie & Thiengo, 2003).

Five species were described as epibionts of

ampullariid mollusks (Haswell, 1893; Ponce

de León, 1979; 1989; Damborenea & Brusa,

2008). Temnocephala iheringi Haswell, 1893

was the first of them, and has several records in

several genera/species of ampullariid mollusks

in Brazil and in Argentina (Pereira & Cuocolo,

1941; Hyman, 1955; Damborenea, 1992;

Damborenea & Cannon, 2001; Damborenea et

al., 1997; Seixas et al., 2010a). Temnocephala

rochensis Ponce de León, 1979 and

Temnocephala haswelli Ponce de León, 1989

were described from Uruguay on Pomacea

canaliculata (Lamarck, 1822) (Ponce de León,

1979; 1989); both had their descriptions

updated when recorded from Brazil by Seixas

et al. (2010b; 2010c). Temnocephala lamothei

Damborenea & Brusa, 2008

Pomella megastoma

(Sowerby, 1825) of Argentina (Damborenea &

Brusa, 2008), and, recently,

has been

described from

Temnocephala

colombiensis Garcés, Puerta, Tabares, Lenis &

Velásquez, 2013, has been described from an

undetermined species of Pomacea Perry, 1810,

from Antioquia, Colombia (Garcés et al.,

2013).

Representatives of Neritidae Lamarck, 1809,

usually are euryhaline, some of them more

associated to the marine environment, while

other species are associated to brackish waters

(Tan & Clements, 2008). Species of Neritina

Rafinesque, 1815, inhabit sandy and muddy

environments, living over pieces of wood and

RESUMO

Palavras-chave: moluscos - Região Neotropical – Taxonomia – temnocefalídeos.

Temnocephala sp. n., ectosimbionte de moluscos da espécie Neritina zebra

(Bruguière, 1792) é descrita do Estado do Pará, Amazônia Brasileira. Representantes da família

Neritidae Lamarck, 1809, geralmente eurialinos, são adaptados aos habitats que apresentam um

certo grau de salinidade. Até o momento, todas as espécies de Temnocephala epibiontes em

moluscos foram descritas/registradas em hospedeiros da família Ampullariidae Gray, 1824,

estritamente dulciaquícola como todos os hospedeiros conhecidos das espécies de

Temnocephala. Vinte e um moluscos foram coletados no Rio Tocantins, Município de Cametá. A

nova espécie é diferente de todas as espécies descritas sobre moluscos até o momento, e possui os

seguintes caracteres: 1. tamanho corporal diminuto; 2. cirro simples e curto, diferente dos cirros

das espécies epibiontes de moluscos que, salvo variações específicas, apresentam o formato de

pincel, onde o limite entre o shaft e o introvert é bem marcado e o último apresenta uma inflação;

3. sistema reprodutor feminino simples e inconspícuo; 4. vagina pouco muscular e sem

diferenciação entre as porções proximal e distal; 5. vesícula intermedia pequena, substituindo os

receptáculos seminais; e 6. esfíncter vaginal único e simétrico. Ovos de temnocefalídeos não

foram encontrados dentro ou fora da concha dos hospedeiros, assim como sobre os opérculos.

Este é o primeiro registro de uma espécie de Temnocephala epibionte em moluscos neritídeos e o

primeiro registro sobre um hospedeiro de hábito eurialino.

euryhalina

Seixas et al.

dorsolateral, post-tentacular 'excretory'

syncytial plates (DLSPs). Photomicrographs

were taken with the microscopes Zeiss Axiolab

and Leica DMR Hc equipped with Nomarski´s

differential interference contrast (DIC) prisms.

The photographic images and line drawings

were scanned and prepared using CorelDraw

X5 and Adobe's Photoshop CC, respectively.

Measurements are in micrometers (µm) unless

otherwise indicated and were taken from

specimens killed under slight cover slip

pressure and mounted in Canada balsam;

ranges are followed (between parentheses) by

the mean, the standard deviation values, and

the number of specimens measured for a given

character (when different than 8). Cirrus

measurements were taken from extracted cirri

mounted in Faure´s mounting medium (F),

while the terminology used to describe the

male reproductive structures followed Seixas

et al. (2010a).

The whole mounts of

adult and juvenile specimens, as well as slides

containing individual cirri mounted in (F)

were

Temnocephala n. sp.

(Figs 3 - 24)

Description. Based on 14 specimens collected:

8 whole mounted adults; 2 specimens mounted

The distribution of

rhabditogenic and disc glands was studied by

clearing juvenile specimens in lactophenol

(Amato & Amato, 2005).

deposited in the following scientific

collections:1. 'Coleção Helmintológica do

Instituto Oswaldo Cruz (CHIOC)', Rio de

Janeiro, RJ, Brazil; 2. 'Colección de

Invertebrados, División Zoología

Invertebrados, Museo de La Plata (MLP)', La

Plata, Argentina; 3. 'Coleção de Invertebrados

do Instituto Nacional de Pesquisas da

Amazônia (INPA)', Manaus, AM, Brazil; and

4. 'Coleção Helmintológica do Laboratório de

Helmintologia, Departamento de Zoologia,

Universidade Federal do Rio Grande do Sul',

Porto Alegre, RS."

euryhalina

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015

other plant remains near the shore. They are

found on submerged areas, never exposed,

always on roots of trees of the river (Barroso &

Matthews-Cascon, 2009). Neritina zebra

(Bruguière, 1792) occurs in the Brazilian

seashore regions (State of Pará to State of Rio

de Janeiro), and in Suriname.

The present work records a new species of

Temnocephala, which is found for the first time

on Neritid mollusks, and is the first record of a

host with euryhaline habitat.

Twenty-one specimens of N. zebra (Figs 1 and

2) were collected in February of 2010 in Rio

Tocantins (2º14'35.39”S, 49º29'50.29”W),

Municipality of Cametá, Pará, Brazil, distant

144 km, in a straight line to Belém, PA. The

mollusks were collected with dip nets by

Edilson R. Mattos and were taken still alive to

the “Laboratório de Pesquisa Carlos Azevedo

– LPCA, Universidade Federal Rural da

Amazônia (UFRA)”, Belém, PA, to be

e x a m i n e d f o r t e m n o c e p h a l a n s .

Temnocephalans were fixed with 10%

phosphate-buffered formalin 90ºC or with

AFA, under slight cover slip pressure,

following the protocols established by Amato

et al. (2007) and Seixas et al. (2010a). Some

specimens were stained in Delafield's

hematoxylin or aceto-carmine /fast green,

cleared in cedar oil, and mounted as permanent

slides in Canada balsam. Other specimens

were prepared for Scanning Electron

Microscopy (SEM) at the 'Centro de

Microscopia Eletrônica da UFRGS (CME)'.

The temnocephalans from N. zebra were

studied through a series of techniques focusing

especially on the: 1. morphology of the cirrus

structure; 2. morphology of the vagina and

other female reproductive organs; 3.

distribution of the rhabditogenic glands in

juveniles; and, 4. the shape of the paired,

MATERIAL AND METHODS

RESULTS

A new species of Temnocephala

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 Seixas et al.

on stubs for SEM; 6 dissected cirri mounted in

(F); 8 specimens measured.

External characteristics. Body (without

tentacles) (Figs 3 and 5) 869–1.36 mm (1.10

mm ± 164) long, 494–691 mm (612 mm ± 64)

wide; adhesive disk ventral, subterminal,

covered by body (Figs 5, 10, and 13 -

ad) 197–336 (247 ± 57) long, 237–454 (355 ±

82) wide; disc peduncle 190–310 (226 ± 39)

wide. ed pigmentation present

(observations made on live specimens). Two

DLSPs, slightly wider than (Figs

partially

Eyespots with r

rounded long

9 and 11-12), left

plate 99–147 (123 ± 34, 2) long, 109.2–109.85

(109 ± 0.45, 2) wide; right plate 107-154 (131

± 33, 2) long, 100–112 (106 ± 8, 2) wide; length

ratio of DLSPs /total body length, without

tentacles, 8: 1. Excretory pore in the anterior

portion of the plate, near the internal limit (Figs

11-12 - ep).

Glands. Rhabditogenic glands forming

bunches (average 13 cells) extending from

external margin reaching

ventrolateral margin of body (Fig. 12),

Figures 1-2. Neritina zebra. (1) Shell, dorsal view. Bar = 2.5 mm. (2) Shell, ventral view, showing the operculum (asterisk) and

the mollusk foot (arrow), still fixed inside the shell. Bar = 2.5 mm.

Figure 3. Adult of Temnocephala , stained with acetic carmine/fast-green and mounted in toto. Bar = 100 µm.euryhalina n. sp.

mid-level of intestine to anterior level of

posterior testes, in lateral fields of body, 25-

42.5 (35 ± 7) in diameter (Fig. 4 – black head

arrow), ducts inconspicuous. Two groups of

two Haswell glands (Fig. 5 - hg), showing little

affinity with hematoxylin, in front of the

cerebral transverse band (Fig. 5 - b), diameter

of largest cell 37.5–52.5 (45 ± 5; 6).

Esophageal glands between pharynx and the

intestine (Fig. 4 - eg). Disc glands between

adhesive disc and genital complex, 27.5–55

(39 ± 8) in diameter, forming two, lateral

bunches extending from mid-level of posterior

testes to anterior margin of adhesive disc,

including two pairs of large, round, more

central paranephrocytes, 50–62.5 (56 ± 4; 5)

long (Figs 4 and 24 - asterisk).

Reproductive system.

Female. Ovary 57.5–117.5 (85 ± 18) long,

57.5–100 (76 ± 15) wide, located between the

vagina and the vesicula resorbens (Figs 6 and

20-21 - o). Vitellarium arborescent and dense

(Figs 3 and 5 - vg); vagina with thick muscular

walls 47.5–97.5 (75 ± 18) long, 22.5-40 (28 ±

6) wide (Figs 6, 20-22 - v); single vaginal

sphincter symmetrical, 25-37.5 (28 ± 5) in total

diameter (Figs 6 and 21-22 – avs/pvs); vesicula

intermedia small, 7.5-15 (11 ± 2) long (Figs 6

and 21 - vi); vesicula resorbens usually full of

sperm, 37.5–100 (60 ± 22) long, 67.5–150 (100

± 26) wide (Figs 6 and 20-22 – vr).

Figure 4. Juvenile of Temnocephala , cleared in lactophenol and mounted in toto, showing esophageal glands

(eg), paranephrocytes (asterisks) and the limit of the rhabditogenic glands (black head arrows). Bar = 50 µm.

euryhalina n. sp.

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 A new species of Temnocephala

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 Seixas et al.

Figures 5-8. Partial diagrams of Temnocephala (5) Adult, ventral view. Mouth (m), adhesive disc (ad), pharynx

(ph), cerebral band (b), Haswell glands (hg), vitelline glands (vg), intestine (i), paranephrocytes (head arrows), tentacles (t),

anterior testes (at), posterior testes (pt), excretory vesicles (ev). Bar = 100 µm. (6) Feminine organs of the reproductive system.

Genital atrium (ga), vitelline gland duct (vgd), anterior vaginal sphincter (avs), posterior vaginal sphincter (pvs), ovary (o),

vesicula intermedia (vi), vagina (v), and vesicula resorbens (vr). Bar = 50 µm. (7) Cirrus, showing the limit between introvert and

shaft (arrow). Bar = 25 µm. (8) Masculine organs of the reproductive system. Prostatic bulb (pb), showing an opening of the wall

(head arrow), cirrus (c), prostatic cells (pc), vasa deferentia (vd), prostatic secretions (ps), ejaculatory vesicle (ejv), and seminal

vesicle (sv). Bar = 50 µm.

euryhalina n. sp.

Figures 9-13. Temnocephala observed with SEM. (9) Adult specimen, showing the limits of the dorsolateral

'excretory' syncytial plates (DLSPs) (head arrows) and the excretory pores (ep). Barra = 100 µm. (10) Posterior region showing

the adhesive disc syncytial plate limits (head arrow), the genital pore (gp), and the secretions produced by the disc glands

(asterisk). Bar = 50 µm. (11) Limits (internal, superior, and inferior) of the right DLSP (head arrows) and the excretory pore (ep).

Bar = 20 µm. (12) Limits (external, superior, and inferior) of the right DLSP (head arrows) and the excretory pore (ep). Bar = 20

µm. (13) Posterior region, detail of the genital pore (gp) and the adhesive disk (ad). Bar = 20 µm.

euryhalina n. sp.,

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 A new species of Temnocephala

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 Seixas et al.

Male. Four testes usually rounded, slightly

oblique; anterior testes always superposed to

posterior testes (Figs 5 and 20 - at/pt), deferent

vessels unite in small, pyriform seminal

vesicle, 70–117.5 (87 ± 17; 6) long, 35–60 (40

± 10; 6) wide (Figs 8, 20, and 23 - sv); prostatic

bulb round and short, 42.5–92.5 (62 ± 19; 7)

long, 42.5–65 (52 ± 79; 7) wide (Figs 8, 20, and

23 - pb). Cirrus short, wide 112.5–140 (124 ±

10; 5) long (Figs 7-8, 14, 20 and 23), introvert

distal portion with spines from tip to mid-level,

followed by smooth region, extending to the

introvert proximal limit (Figs 7 and 15-19).

Shaft 77.5–115 (90 ± 14; 5) long, shaft

Figures 14-19. Cirri of Temnocephala , photomicrographed with differential interference contrast (DIC) with

Nomarski's prisms. (14) Entire cirrus, showing the limit between the shaft and the introvert (arrow). Bar = 25 µm. (15-19) Cirrus

introvert observed in a sequence of several focusing planes, showing the limit between the shaft and the introvert (arrow). Bar =

20 µm.

euryhalina n. sp.

Figures 20-24. Temnocephala (20) Organs of the reproductive system. Genital atrium (ga), prostatic bulb (pb),

cirrus (c), vaginal sphincter (vs), ovary (o), genital pore (gp), anterior testes (at) slightly superposed to the posterior testes (pt),

vesicula intermedia (asterisk), vagina (v), seminal vesicle (sv) and vesicula resorbens (vr). Bar = 100 µm. (21) Organs of the

feminine reproductive system. Anterior vaginal sphincter (avs) and posterior vaginal sphincter (pvs), ovary (o), genital pore (gp),

vagina (v), vesicula intermedia (vi), and vesicula resorbens (vr). Bar = 50 µm. (22) Organs of the feminine and part of the

masculine reproductive systems. Genital atrium (ga), cirrus (c), anterior vaginal sphincter (avs) and posterior vaginal sphincter

(pvs), genital pore (gp), vagina (v), vesicula intermedia (asterisk), and vesicula resorbens (vr). Bar = 50 µm. (23) Organs of the

masculine reproductive system. Prostatic bulb (pb), cirrus (c), prostatic cells (pc), prostatic secretions (ps), anterior testes (at)

slightly superposed to the posterior testes (pt), ejaculatory vesicle (asterisk), and seminal vesicle (sv). Bar = 50 µm. (24) Posterior

region showing the posterior testis (pt) and the paranephrocytes (asterisks). Bar = 100 µm.

euryhalina n. sp.

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 A new species of Temnocephala

DISCUSSION

Temnocephala n. sp. is the smallest

among the five species from mollusks known

to date, having an average body length of 1.10

mm. Temnocephala iheringi, T. rochensis, T.

haswelli, and T. lamothei are, in average, 2.03-

2.80 mm long ( , while T.

colombiensis, the smallest species described to

date, has, in average a body length of 1.62 mm

(Garcés et al., 2013).

Temnocephala lamothei is the most similar

species to the epibionts from N. zebra,

presenting a reproductive system less complex

than those of the three other species described

from mollusks, notwithstanding, the body size,

the female reproductive system, and the cirrus

are larger than in T. Another

important aspect of T. lamothei is the introvert

which does not show an inflation area, but

shows a widening similar to a ring

(Damborenea & Brusa, 2008).

The specimens from N. zebra have unique

characteristic among the epibiont species from

mollusks; the cirrus of T. is

simple and short, while in epibionts of other

species from mollusks, except for intraspecific

variations, it has the shape of a brush (

, where the limit between the

shaft and the introvert is well marked and the

later has an inflation area. The female

reproductive system is simple and

inconspicuous, different from the feminine

organs in the other species from

ampulariideans (

. The vagina of T. e is not

very muscular (Figs 6 and 20-22 - v) and has no

differentiation between the proximal and distal

portions, besides having a single and

symmetrical vaginal sphincter (Fig. 6 -

avs/pvs).

Temnocephala euryhalina is the only

euryhalina

euryhaline n. sp.

euryhalina n. sp.

uryhalinan. sp.

n. sp.

Damborenea & Brusa, 2008;

Seixas et al. 2010a; 2010b; 2010c)

Seixas et

al. 2010a; 2010b)

Seixas et al. 2010a; 2010b;

2010c)

maximum width at base 57.5–75 (64 ± 7; 5);

introvert 25-37.5 (33 ± 5; 5) long; introvert

width at base 22.5-27.5 (25 ± 2, 5); maximum

introvert width at level of swelling 30-40 (32 ±

4, 5). Introvert´s swelling with approximately

28 rows of spines, and 5 spines long and thin in

each row (Figs 15-19). Ratio between total

body length (without tentacles)/ total length of

cirrus 8.8: 1; ratio between total length of

cirrus/ maximum width of shaft´s base 2: 1;

ratio between total length of cirrus/ total length

of introvert 4: 1.

Taxonomic summary.

Type host: Neritina zebra (Bruguière, 1792)

(Gastropoda, Neritaemorphi, Neritidae).

Type locality: Rio Tocantins (2º14'35.39”S,

49º29'50.29”W), Cametá, Pará, Brazil.

Site of infestation:

Prevalence: 71.42%.

Other helminth specimens examined:

Temnocephala iheringi, Temnocephala

haswelli, and Temnocephala rochensis from P.

canaliculata – voucher specimens deposited in

the 'Coleção Helmintológica do Laboratório de

Helmintologia, Departamento de Zoologia,

Universidade Federal do Rio Grande do Sul';

Temnocephala iheringi - MLP 3118, 3119,

3121, and 3120 deposited in the 'Colección de

Invertebrados, División Zoología

Invertebrados, Museo de La Plata (MLP)'.

Adults and juveniles in

mantle cavity, eggs not found in the body, on

the shell, and neither on the operculum.

Helminth specimens deposited: '

CHIOC 38046 (SBA 3199-1-1 - Holotype);

CHIOC 38047 (SBA 3216-1-1 - cirrus).

'INPA

661 (SBA 3199-1-4 - Paratype); INPA 662

(SBA 3214-1-3 - cirrus) Colección de

Invertebrados, División Zoología

Invertebrados, Museo de La Plata': MLP-He

6842 (SBA 3199-1-3 - Paratype; SBA 3214-1-

4 - cirrus).

Coleção

Helmintológica do Instituto Oswaldo Cruz':

Coleção de Invertebrados do Instituto

Nacional de Pesquisas da Amazônia':

. '

Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 Seixas et al.

Laboratório de Anatomia Vegetal, Instituto de

Biociências, UFRGS, for the permission to use

the Leica DMR HC microscope to make the

DIC photomicrographs. We are indebted to

Edilson Rodrigues Matos and his students

Diego and Neto, at the “Laboratório de

Pesquisa Carlos Azevedo – LPCA,

Universidade Federal Rural da Amazônia

(UFRA)” for the collection of the mollusk

hosts. Special thanks are also due to the staff of

the “Centro de Microscopia Eletrônica”,

UFRGS for the SEM operation; to Maria

Cristina Damborenea, Curator, Invertebrate

Collection at Museo de La Plata (MLP), for the

loan of some Argentinean specimens deposited

at that Institution; to CAPES (Coordenação de

Aperfeiçoamento do Pessoal de Nível

Superior) for the Doctoral Scholarship

awarded to SAS (2008-2013); to Abner

Chiquieri for his contribution to the correct

formation of the specific epithet of the new

species; to Débora N. Souza and Lucas

Casagrande for their help in the laboratory;

and, especially, to Philip J. Scholl, for kindly

reviewing the English in the several versions of

the manuscript.

species epibiont on mollusks with vesicula

intermedia (Figs 6 and 21 - vi). Temnocephala

lamothei has only one, instead of four seminal

receptacles (Damborenea & Brusa, 2008). The

vesicula intermedia is only a widening of the

wall while seminal receptacle is a single

vesicle located just after the ootype. All the

other species epibiont on mollusks have four,

conspicuous seminal receptacles. Seixas et al.

(2010c) when describing specimens of T.

rochensis, from Rio Grande do Sul, mistakenly

called vesicula intermedia the proximal

portion of the vagina, although the species has

four seminal receptacles and the distal portion

has a very strong musculature, which

differentiates it from the proximal portion.

The testes, usually large and oblique in the

epibionts from mollusks, are small, rounded,

of similar size, and always superposed in the

specimens of T. e (Figs 5, 20,

and 23 - at/pt).

The shells of ampulariideans infested with

temnocephalans usually show a large number

of eggs in the spire and in the body whorl of the

shell ( , as

well as the operculum (Damborenea & Brusa,

2008). Shells and opercula of N. zebra were

painstakingly examined for eggs, however, not

a single temnocephalan egg was found in any

of the collected mollusks. The euryhaline habit

of the host, promotes the contact of the

temnocephalans with a certain degree of

salinity, which opens the possibility that this

species lays the eggs in the interior portion of

the shell. However, more than 90% of the

collected shells (some shells were kept as

testimonials) were broken in the search for

internal postures, and not a single egg has been

found.

Special thanks are due to Jorge Ernesto de

Araújo Mariath and Rinaldo Pires dos Santos,

uryhalina n. sp.

Seixas et al., 2010a; 2010b; 2010c)

ACKNOWLEDGMENTS

Amato, JFR & Amato, SB. 2005.

Amato, JFR, Seixas, SA & Amato, SB. 2007.

New species

o f Te mn oc e ph a la B l an c ha rd

(Platyhelminthes, Temnocephalida)

ectosymbiont on giant water bugs,

Bel o st om a spp . (H e m ip te r a,

Belostomatidae) from southern Brazil.

Revista Brasileira de Zoologia, vol. 22,

pp. 107–118. A

new species of Temnocephala Blanchard

(Platyhelminthes, Temnocephalida)

ectosymbiont on creeping water bugs,

Cryphocricos granulosus De Carlo

(Hemiptera, Naucoridae) from southern

Brazil. Revista Brasileira de Zoologia,

vol. 24, pp. 1043-1051.

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Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015 A new species of Temnocephala