A COMPLETE DESCRIPTION OF THE FEMALE REPRODUCTIVE SYSTEM OF
TEMNOCEPHALA CYANOGLANDULA AMATO, AMATO & DAUDT
(PLATYHELMINTHES,
TEMNOCEPHALIDA)
DESCRIÇÃO COMPLEMENTAR DO SISTEMA REPRODUTOR FEMININO DE
TEMNOCEPHALA CYANOGLANDULA AMATO, AMATO & DAUDT
(PLATYHELMINTHES, TEMNOCEPHALIDA)
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(2), jul-dec: 371-376.
ABSTRACT
Keywords: Aeglidae - ectosymbiont - Neotropical Region - South America - taxonomy - Temnocephala cyanoglandula.
Temnocephala cyanoglandula Amato, Amato & Daudt, 2003 was described as epibiont of Aegla
serrana Buckup & Rossi using the usual techniques at the time of publication, but did not include
the description of the female reproductive system. A complete description of the female
reproductive system was accomplished based on specimens stained with acetic carmine/fast-
green for better observation of the female organs and with the aid of Nomarski's differential
interference contrast (DIC). The taxonomic validation of T. cyanoglandula, was questioned
because of the similarity between the cirrus of this species and that of Temnocephala axenos
Monticelli, 1899 and Temnocephala bresslaui Pérez-González, 1949 (synonymized with T.
axenos), has subsequently been confirmed. The cirrus in T. cyanoglandula is much longer than
that of T. axenos, and, although there is similarity of the cirrus with that of T. bresslaui, the later
has much longer female organs and is probably a valid species. The female reproductive organs
of T. cyanoglandula and Temnocephala mertoni Volonterio, 2007 are similar, although both
species have cirri of different size and shape.
371
ISSN Versión impresa 2218-6425 ISSN Versión Electrónica 1995-1043
RESEARCH NOTE/NOTA CIENTÍFICA
INTRODUCTION
372
interference contrast (DIC) in all descriptions,
mainly in the species epibiont on crustacean
hosts. The proposed protocol was followed in
t h e d e s c r i p t i o n o f Te m no c eph a la
longivaginata Seixas, Amato & Amato, 2011,
species which, although having a large body
size, had a very small, difficult to observe
female reproductive system (Seixas et al.,
2011). The description of T. cyanoglandula,
correct in relation to the male organs
(principally the cirrus) and the dorsolateral
'excretory' syncytial plates, did not include the
description of the female reproductive system.
For this reason these structures have now been
stained and observed by the techniques
proposed by Amato et al. (2010) and are
described in the present note.
The specimens described in the original
manuscript (Amato et al. 2003), were collected
at Utopia II Farm, 5 km East of State Road RS-
020, locality of Tainhas, Municipality of
Cambará do Sul, State of Rio Grande do Sul
RESUMO
Palavras-chave: Aeglidae - América do Sul - ectosimbionte - Região Neotropical - taxonomia.
Temnocephala cyanoglandula Amato, Amato & Daudt, 2003 foi descrita como epibionte de
Aegla serrana Buckup & Rossi, 1977 utilizando as técnicas propostas até o momento da
publicação, por essa razão não continha a descrição do sistema reprodutor feminino. A descrição
complementar do sistema reprodutor feminino foi feita baseada em espécimes corados com
carmin acético/ fast-green e da visualização dos órgãos femininos em contraste diferencial de
interferência com prismas de Nomarski (DIC). A validade taxonômica de T. cyanoglandula,
questionada pelas semelhanças entre os cirros desta espécie com Temnocephala axenos
Monticelli, 1899 e Temnocephala bresslaui Pérez-González, 1949 (sinonimizada como T.
axenos), foi subsequentemente confirmada. O cirro de T. cyanoglandula é muito maior em
comprimento do que T. axenos e, apesar da semelhança do cirro com T. bresslaui, a última
apresenta as estruturas femininas muito longas e é, provavelmente, uma espécie válida. Os órgãos
reprodutores femininos de T. cyanoglandula e Temnocephala mertoni Volonterio, 2007 são
semelhantes, no entanto possuem cirros de tamanho e formato diferentes.
Temnocephala cyanoglandula Amato, Amato
& Daudt, 2003 has been described as epibiont
of Aegla serrana Buckup & Rossi, 1977
following the techniques proposed by Cannon
& Sewell (1995), Sewell & Cannon (1998),
and Amato et al. (2003). The female
r e pr o d u c t iv e o rg a n s f o u nd i n t h e
temn oc eph al an s pe ci es e pi bi ont on
crustaceans are very difficult to observe.
Volonterio (2007), comparing Temnocephala
mertoni Volonterio, 2007, Temnocephala
talicei Dioni, 1967, and Temnocephala axenos
Monticelli, 1899 called attention to the fact
that due to the difficulty in differentiating
species which share the same host group or
even the same host species, organs such as the
vaginal sphincter have the same, or even
greater importance than the description of the
cirrus. Amato et al. (2010) when recording the
presence of Temnocephala pignalberiae
Dioni, 1967 in Brazil, proposed to use the
acetic carmine/fast-green stain combination
and the observation of the female reproductive
system with Nomarski's differential
MATERIAL AND METHODS
Neotropical Helminthology. Vol. 9, Nº2, jul-dec 2015 Seixas et al.
(29º15'10”S, 50º15'45”W), southern Brazil,
were used in the present study. These
o
specimens were fixed in cold AFA (70 GL
ethanol; formalin 37%; glacial acetic acid),
and stained in aceto-carmine/fast-green. The
female organs were extracted and mounted in
Faure´s mounting medium (F) (Seixas et al.,
2010). Photomicrographs were taken with
Zeiss Axiolab and Leica DMR Hc
microscopes; the Leica microscope was
equipped with DIC. The photographic images
were prepared using Adobe's Photoshop CC.
Measurements are in micrometers (μm) 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 10). The whole mounts of adult, as well as
slides containing individual female organs
mounted in F were deposited in the following
s ci e nt i fi c c ol l ec t io n s: 1 . ' C ol e çã o
Helmintológica do Instituto Oswaldo Cruz
(CHIOC)', Rio de Janeiro, RJ, Brazil; and 2.
'Coleção Helmintológica do Laboratório de
Helmintologia, Departamento de Zoologia,
Universidade Federal do Rio Grande do Sul',
Porto Alegre, RS.
Description. Based on 31 specimens; 10
measured. Female reproductive system. Ovary
almost round (Figs 1, 2, and 4) 112.5-202.5
(144, 29) long, 100-130 (115, 10) wide;
vesicula intermedia (Figs 1, 3, and 5) 20-62.5
(41, n = 9, 13) long; vagina (Figs 3 and 5) 35-
87.5 (55, n = 9, 16) long, 37.5-65 (50, n = 8, 10)
maximum width; single vaginal sphincter
large and asymmetrical (Figs 3 and 5) 50-100
(78, 17) in total diameter, diameter of anterior
portion 12.5-27.5 (21, 6), diameter of posterior
portion 32.5-75 (47, 13); vesicula resorbens
(Figs 1 and 5) 95-290 (181, n = 6, 64) long,
373
Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015
RESULTS
120-310 (222, n = 6, 64) wide.
Taxonomic summary.
Type host: Aegla serrana Buckup & Rossi,
1977 (Crustacea, Anomura, Aeglidae).
Type locality: Fazenda Utopia II, 5 km RS-
020, Tainhas, Cambará do Sul, Rio Grande do
Sul, Brazil.
Site of infestation: Branchial chambers and
body surface; eggs cemented over external
surfaces of exoskeleton, in different regions of
the ventral side: perioral area, pleural strips,
sternal plates, pereiopods, and chelipods; to a
lesser extent on the dorsal side of
cephalothorax and dorsal side of uropods.
Helminth specimens deposited: 'Coleção
Helmintológica do Instituto Oswaldo Cruz':
CHIOC 38214 a - d.
Volonterio (2007) created doubts about the
taxonomic validity of T. cyanoglandula, when
she indicated similarity among the cirri of T.
cyanoglandula, T. axenos, and Temnocephala
b re s s l a u i P é r e z - G o n z á l e z , 1 9 4 9
(synonymized as T. axenos by Baer (1931))
(256 µm total length in average - T.
cyanoglandula, 141 µm total length in average
- T. axenos, and 280 µm total length in average
- T. bresslaui). The measurements of the cirrus,
prostatic bulb, and body size are similar
between T. cyanoglandula and T. bresslaui,
although, Pérez-González (1949) drew and
described the female reproductive system with
peculiar measurements, which according to
Volonterio (2007), did not agree with the
measurements of any species already
described. Temnocephala bresslaui has
vesicula intermedia, called 'ootype' by the
author, with an average of 243 µm in length,
and the vagina has a thick muscular wall which
measures an average of 250 µm in length and
150 µm in width, measurements that exclude
the possibility of synonymy with T. axenos,
DISCUSSION
Complementary description of Temnocephala cyanoglandula
374
whose cirrus is much smaller in length than
that of T. cyanoglandula. Pérez-González
(1949) also has observed seminal receptacles
in live specimens and considered these
structures as transitory. In T. cyanoglandula
the vesicula intermedia can be observed in all
specimens. Temnocephala bresslaui is
probably, a valid species and the re-description
of the type specimens is necessary. When the
morphometric data of the female organs of T.
cyanoglandula are compared with those of T.
mertoni, it is possible to see similarities
Figures 1-3. Temnocephala cyanoglandula. (1) Organs of the reproductive system. Anterior portion of the vaginal sphincter
(avs), genital atrium (ga), prostatic bulb (pb), cirrus (c), ovary (o), genital pore (gp), posterior portion of the vaginal sphincter
(pvs), vesicula intermedia (vi), vagina (v), seminal vesicle (sv), vasa deferentia (vd), and vesicula resorbens (vr). Scale bar = 100
µm. (2-3) Organs of the female reproductive system. Scale bars = 50 µm. (2) Anterior portion of the vaginal sphincter (avs),
genital atrium (ga), ovary (o), posterior portion of the vaginal sphincter (pvs), vesicula intermedia (*), vagina (v), and vesicula
resorbens (vr). (3) Anterior portion of the vaginal sphincter (avs), genital atrium (ga), ovary (o), posterior portion of the vaginal
sphincter (pvs), vesicula intermedia (vi), vagina (v), and vesicula resorbens (vr).
Neotropical Helminthology. Vol. 9, Nº2, jul-dec 2015 Seixas et al.
375
between them. Both species have only one
asymmetric vaginal sphincter which is smaller
in total diameter in T. mertoni, measuring an
average of 49 µm (78 µm in T. cyanoglandula).
According to Volonterio (2007), the majority
of the specimens of T. mertoni have vesicula
intermedia, as the author also described for T.
axenos, the structure is transitory and only
10% have seminal receptacles. The vesicula
intermedia has been observed in all specimens
of T. cyanoglandula, as a relatively large
structure (Figs 3 and 5). The cirrus of T.
mertoni is smaller in length (138 µm in
average) and has a different shape, straight
with a slight curve (described as 'sinuous') in
the distal portion of the shaft, while in T.
cyanoglandula the entire cirrus is slightly
curved (Fig. 1).
Special thanks are due to Jorge Ernesto de
Araújo Mariath and Rinaldo Pires dos Santos,
Laboratório de Anatomia Vegetal, Instituto de
Biociências, UFRGS, for the permission to use
the Leica DMR HC microscope to make the
DI C p h ot om ic ro g ra ph s; t o C A PE S
(Coordenação de Aperfeiçoamento do Pessoal
de Nível Superior) for the Doctoral
Scholarship awarded to SAS (2008-2013); to
Luiz Carlos Campos Daudt for the collection
and exam of the hosts; 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.
Figures 4 and 5. Organs of the female reproductive system of Temnocephala cyanoglandula photomicrographed with
differential interference contrast (DIC) with Nomarski's prisms. Scale bars = 50 µm. (4) Anterior portion of the vaginal sphincter
(avs), ovary (o), posterior portion of the vaginal sphincter (pvs), vesicula intermedia (*), vagina (v), and vesicula resorbens (vr).
(5) Anterior portion of the vaginal sphincter (avs), ovary (o), posterior portion of the vaginal sphincter (pvs), vesicula intermedia
(vi), vagina (v), and vesicula resorbens (vr).
ACKNOWLEDGMENTS
Neotropical Helminthology. Vol. 9, Nº1, jan-jun 2015
Complementary description of Temnocephala cyanoglandula
376
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