Volume11,Number1(ene-jun2017)
ÓrganooficialdelaAsociaciónPeruanadeHelmintologíaeInvertebradosAfines(APHIA)
Lima-Perú
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Neotropical Helminthology, 2017, 11(1), jan-jun: 61-67.
ORIGINAL ARTICLE / ARTÍCULO ORIGINAL
LARVAE OF DIOCTOPHYME RENALE (GOEZE, 1782) (NEMATODA: ENOPLIDA) AND
CONTRACAECUM SP. (NEMATODA: ANISAKIDAE) IN FRESHWATER TURTLES
(TESTUDINES: CHELIDAE) FROM SOUTHERN BRAZIL
LARVAS DE DIOCTOPHYME RENALE (GOEZE, 1782) (NEMATODA: ENOPLIDA) Y
CONTRACAECUM SP. (NEMATODA: ANISAKIDAE) EN TORTUGAS DE AGUA DULCE
(TESTUDINES: CHELIDAE) DEL SUR DE BRASIL
Laboratório de Parasitologia de Animais Silvestres (LAPASIL), Instituto de Biologia,
Universidade Federal de Pelotas (UFPel).
*Correspondence should be sent to: phrybio@hotmail.com
*
Carolina Silveira Mascarenhas & Ana. Beatriz Devantier Henzel & Gertrud Müller
ABSTRACT
Dioctophyme renale (Goeze, 1782) has been reported in several species of wild carnivores and in
domestic dogs. Aquatic oligochaetes act as intermediate hosts, frogs and fish act as paratenic hosts.
Contracaecum spp. parasites fish-eating birds and mammals associated with aquatic environments act as
intermediate hosts, and fish act as intermediate and paratenic hosts. This work aims to report the
occurrence larvae of D. renale and Contracaecum sp. in freshwater turtles. Nine specimens of Phrynops
hilarii (of Duméril & Bibron, 1835) and 21 specimens of Acanthochelys spixii (Duméril & Bibron, 1835)
were collected from Rio Grande do Sul, Brazil. Four specimens of P. hilarii were infected by third-stage
larvae of D. renale and one specimen of A. spixii was infected by third-stage Contracaecum sp. larvae.
This is the first record of larvae of D. renale and Contracaecum sp. in P. hilarii and A. spixii, respectively.
Neotropical Helminthology
61
Keywords: Acanthochelys spixii – giant kidney worm – nematodes – Phrynops hilarii – third-stage larvae
Neotropical Helminthology, 2017, 11(1), jan-jun
RESUMEN
Palabras clave: Acanthochelys spixii – gusano gigante del riñón – larvas de tercer estadio – nematodos – Phrynops hilarii
Dioctophyme renale (Goeze, 1782) se ha reportado en varias especies de carnívoros silvestres y en perros
domésticos. Los oligoquetos acuáticos son hospedadores intermedios, ranas y los peces actúan como
hospedadores paraténicos. Contracaecum spp. es parásito de aves y mamíferos que alimentan de peces,
invertebrados actúan como hospedadores intermedios, y los peces actúan como hospedadores
intermediarios y paraténicos. Este trabajo tiene como objetivo registrar larvas de D. renale y
Contracaecum sp. en tortugas de agua dulce. Se examinaron nueve Phrynops hilarii (Duméril & Bibron,
1835) y 21 Acanthochelys spixii (Duméril & Bibron, 1835), colectados in Rio Grande do Sul, Brasil.
Cuatro ejemplares de P. hilarii estuvieron infectados por larvas de tercer estadio de D. renale y una A.
spixii estuvo infectada por larvas de tercer estadio de Contracaecum sp. Este es el primer registro de larvas
de D. renale y Contracaecum sp. en P. hilarii y A. spixii, respectivamente.
INTRODUCTION which are found, generally, free or encapsulated in
the cavity (Anderson, 2000).
Dioctophyme renale and some species of
Contracaecum have recognized zoonotic potential,
since several cases have been reported in humans,
mainly due to fish consumption (Eiras et al., 2015).
Dioctophyme renale was related to skin and
kidneys infection in humans from Asia (Hanjani et
al., 1968; Urano et al., 2001; Sardjono et al., 2008;
Katafigiotis et al., 2013; Tokiwa et al., 2014).
Anisakiasis in humans may be caused by
Contracaecum spp. and other species of
Anisakidae and there are several cases in the world
(Eiras et al., 2015). Even though here are not cases
in Brazil, there are records of Contracaecum larvae
in several species of fish allowed for human
consumption, such as Hoplias malabaricus (Bloch,
1794), Hoplerythrinus unitaeniatus (Spix &
Agassiz, 1829), Salminus brasiliensis (Cuvier,
1816) and Rhamdia quelen (Quoy & Gaimard,
1824) (Martins et al., 2005; Barros et al., 2007;
Madi & Da Silva, 2009; Mesquita et al., 2014).
Phrynops hilarii (Duméril & Bibron, 1835)
(Chelidae) occurs in Uruguay, Paraguay, Argentina
and Brazil (Santa Catarina and Rio Grande do Sul)
(Djik et al., 2014). This specie is considered one of
the most abundant in the Rio Grande do Sul, being
commonly observed in urban environments, as
well as in the countryside (Bujes & Verrastro,
2008). Acanthochelys spixii (Duméril & Bibron,
Dioctophyme renale (Goeze, 1782) (Nematoda:
Enoplida) has been registered in wild carnivores
and domestic dogs (definitive hosts), parasitizing
the right kidney and, occasionally, both kidneys
and the peritoneal cavity (Anderson, 2000). The
life history of D. renale was studied in North
America, where fishes and amphibians act as
paratenic hosts when they become infected by
ingesting aquatic oligochaetes (intermediate hosts)
with third-stage larvae (Mace & Anderson, 1975;
Measures & Anderson, 1985). Third-stage D.
renale larvae were reported in frogs, fish, and
freshwater turtles in Brazil (Pedrassani et al., 2009;
Abdallah et al., 2012; Mascarenhas & Müller,
2015a; Mascarenhas et al., 2016). However, there
is a large gap in the knowledge about the life cycle
of D. renale, despite several reports of the parasitic
helminth in domestic dogs and wild animals
(Mascarenhas & Müller, 2015a).
Contracaecum spp. (Nematoda: Anisakidae)
occurs in the stomach and small intestine of birds
and mammals piscivorous (definitive hosts). The
eggs of Contracaecum species are eliminated in the
definitive hosts feces, and the development of the
second-stage larvae free occurs in the water. Later
on, it is ingested by invertebrates or fishes
(intermediated hosts), in which occurs the
development of the third-stage larvae infective,
Mascarenhas et al.
62
63
1835) (Chelidae) can be found in Uruguay,
Argentina and Brazil (Djik et al., 2014). In Brazil,
it occurs in the states of Bahia, Goiás, Minas
Gerais, São Paulo, Paraná, Santa Catarina, Rio
Grande do Sul, and in the Federal District (Brasil et
al., 2011; Djik et al., 2014). In the state of Rio
Grande do Sul, this species is associated with sandy
lagoon environments, wetlands and seasonal or
semi-permanent ponds (Bujes & Verrastro, 2008).
In this paper, we report for the first time the
occurrence of third-stage D. renale larvae in
freshwater turtle P. hilarii and third-stage
Contracaecum sp. larvae in A. spixii.
Thirty freshwater turtles, nine specimens of P.
hilarii and 21 of A. spixii, were examined, which
were dead run over victims on highways in the
southern of the State of Rio Grande do Sul, Brazil.
The cities were Pelotas (31°46'19”S -
52°20'33”W), Capão do Leão (31°46'30”S -
52°26'55”W), Rio Grande (32°10'60”S -
52°50'55”W) and Santa Vitória do Palmar
(33°32'20”S - 53°20'59”W). The collections were
made under license from the Instituto Chico
Mendes de Conservação da Biodiversidade
(ICMBio/n° 47397). From the total of examined
turtles, seven were donated by Núcleo de
Reabilitação da Fauna Silvestre (NURFS-UFPel)
after their death during the rehabilitation process.
Turtles were necropsied, examined and nematodes
were fixed in AFA (70°GL ethanol, formalin 37%
and glacial acetic acid), preserved in 70°GL
glycerinated alcohol, cleared and mounted in
Amann's lactophenol (phenol, lactic acid, glycerin
and water). Specimens were identified according to
Mace & Anderson (1975), Measures & Anderson
(1985), Martins et al. (2005) and Pedrassani et al.
(2009). Vouchers were deposited in the “Coleção
de Helmintos do Laboratório de Parasitologia de
Animais Silvestres (CHLAPASIL/UFPel)”
(Numbers 680 and 681), Rio Grande do Sul State,
Brazil. Parasitological indexes followed Bush et al.
(1997). Photomicrographs were prepared in the
microscope Olympus BX 41 with a camera system
and the plates were made in the Adobe
PhotoshopCS5.
Neotropical Helminthology, 2017, 11(1), jan-jun
Dioctophyme renale and Contracaecum sp. in freshwater turtles
MATERIALS AND METHODS
Four specimens of P. hilarii were infected by third-
stage larvae of D. renale (Fig. 1) with one larva per
cyst occurring in the muscles, body cavity (surface
of esophagus, stomach, lung, liver) and serous of
stomach. The mean intensity (MI) was 3.75
helminths/host (1-5 larvae). Third-stage larvae of
Contracaecum sp. (Fig. 2) occurred in one
specimen of A. spixii with six helminths.
The first reported of third-stage D. renale larvae in
freshwater turtle was in Trachemys dorbigni
(Duméril & Bibron, 1835) (Testudines: Emidydae)
(n=32) with prevalence of 87.5% and mean
intensity of infection of 13.9 (Mascarenhas &
Müller, 2015a,b). This report was in urban area
from Pelotas, where also was reported third-stage
D. renale larvae in fish, Hoplosternum littorale
(Hancock, 1828) (Siluriformes: Callichthyidae),
with 4.5 helminths/host (1-5 larvae), four hosts
parasited (Mascarenhas et al., 2016). Mascarenhas
& Müller (2015a) suggests that the significant
prevalence of larvae in turtles in the city of Pelotas
might be related to characteristics of the urban
zone, where to the presence of parasitized domestic
dogs, which spread eggs through their urine; hence,
contaminating urban water bodies that turtles and
oligochaetes cohabit. Additionally, the process of
eutrophication caused by the demand for organic
matter and other household, wasted in these aquatic
urban environments, may have favored an
increased oligochaeta population, contributing to
maintenance of the D. renale cycle (Mascarenhas
& Müller, 2015a). High densities of oligochaetes
are usually found in altered aquatic environments
due to an increase in food resources and a decrease
in oxygen supply, and the ensuing decrease of
predators and/or competitors (Martins et al., 2008).
Dogs population in Pelotas is approximately
66,723 (46,706 semi-domestic, 6,672 street dogs
and 13,345 domestic dogs) (Municipal
Administration, 2012). Therefore, the report of
third-stage D. renale larvae in freshwater turtle and
fish in urban areas of Pelotas is an alert for canine
dioctophymatosis.
About larvae of Contracaecum, Foster et al. (1998)
examined 41 specimens of Apalone ferox
(Schneider, 1783) (Testudines: Trionychidae) in
Southeastern Florida (USA) and registered larvae
RESULTS AND DISCUSSION
64
Neotropical Helminthology, 2017, 11(1), jan-jun Mascarenhas et al.
Figure 1. Third-stage larva of Dioctophyme renale parasite of Phrynops hilarii (Duméril & Bibron, 1835) (Testudines:
Chelidae) from southern Brazil. A – Anterior extremity of female larva (oe – oesophagus) (Bar = 62 µm). B – Posterior
extremity of male larva (Bar = 37 µm). C – Genital primordium (arrow) of female larva near to junction oesophagus-intestine
(oe – oesophagus, in – intestine) (Bar = 290 µm). D – Detail of genital primordium (arrows) and intestine (in) of female larva
(Bar = 290 µm).
of Contracaecum in the stomach of 29% hosts with
mean intensity of 13 (1-66 larvae), also related to
one adult male of Contracaecum multipapillatum
(Drasch, 1882) in the small intestine of one host.
Foster et al. (1998) suggest that infection of turtles
was by ingestion of freshwater fish, which are part
of the species diet. Similarly, the infection of A.
spixii can occurs by ingestion of fish, since this
species also uses fish as a food source
(Huckembeck et al., 2007). This is the first record
of Contracaecum larvae in freshwater turtle, A.
spixii.
The role of P. hilarii and A. spixii in cycle of D.
renale and Contracaecum sp. is questionable,
because the freshwater turtles are not easily
predator by definitive hosts of these helminths. In
relation to D. renale, it is emphasized that P. hilarii
examined, in this study, were found dead on
highways in the urban area, which can facilitate a
possible predation by stray dogs, which have poor
selective eating habits. Therefore, further studies
are needed to understand the life cycle of D. renale
in the region, for the importance of this parasite in
public health.
65
Neotropical Helminthology, 2017, 11(1), jan-jun
Dioctophyme renale and Contracaecum sp. in freshwater turtles
Figure 2. Third-stage larva of Contracaecum sp. parasite of Acanthochelys spixii (Duméril & Bibron, 1835) from southern
Brazil. A – Anterior extremity of larva (oe – oesophagus, ic – intestinal caecum) (Bar = 87 µm). B – Detail of ventricular appendix
(va) and intestine (in) (Bar = 85 µm).
66
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Special thanks to “Núcleo de Reabilitação da
fauna silvestre e Centro de Triagem de Animais
Silvestres da Universidade Federal de Pelotas
(NURFS/CETAS-UFPel), Marco Antônio A.
Coimbra, Fabiana F. Bernardon, Bruna M. Chaviel
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was supported by CAPES (Coordenação de
Aperfeiçoamento do Pessoal de Nível Superior,
process number 32/2010).
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Dioctophyme renale and Contracaecum sp. in freshwater turtles
Received December 6, 2016.
Accepted January 15, 2017.