8
Neotrop. Helminthol., 5(1), 2011
2011 Asociación Peruana de Helmintología e Invertebrados Afines (APHIA)
ISSN: 2218-6425 impreso / ISSN: 1995-1043 on line
Resumen
Abstract
ORIGINAL ARTICLE / ARTÍCULO ORIGINAL
CLINOSTOMUM MARGINATUM (BRAUN, 1899) AND AUSTRODIPLOSTOMUM COMPACTUM
(LUTZ, 1928) METACERCARIAE WITH ZOONOTIC POTENCIAL ON PYGOCENTRUS
NATTERERI (KNER, 1858) (CHARACIFORMES: SERRASALMIDAE) FROM CENTRAL
AMAZON, BRAZIL
CLINOSTOMUM MARGINATUM (BRAUN, 1899) Y AUSTRODIPLOSTOMUM COMPACTUM
(LUTZ, 1928) METACERCARIAS CON POTENCIAL ZOONÓTICO EN PYGOCENTRUS
NATTERERI (KNER, 1858) (CHARACIFORMES: SERRASALMIDAE) DE LA AMAZONÍA
CENTRAL, BRASIL
1,2 1 2 1
Aprigio Mota Morais , Angela Maria Bezerra Varella , Berenice M.M. Fernandes & José Celso de Oliveira Malta
1 2
INPA- Instituto Nacional de Pesquisas da Amazônia, Laboratório de Parasitologia e Patologia de Peixes, Manaus, AM. Bolsista FAPEAM-
3
Fundação de Amparo à Pesquisa do Estado do Amazonas IOC-FIOCRUZ, RJ- Instituto Oswaldo Cruz, Laboratório de Helmintos Parasitos
de Peixes.
Suggested citation: Morais, A.M., Varella, A.M.B., Fernandes, B.M.M. & Malta, J.C.O. 2011. Clinostomum marginatum (Braun, 1899) and
Austrodiplostomum compactum (Lutz, 1928) metacercariae with zoonotic potencial on Pygocentrus nattereri (Kner, 1858) (Characiformes:
Serrasalmidae) from Central Amazon, Brazil. Neotropical Helminthology, vol. 5, n° 1, pp. 08-15.
The present study investigated the presence of helminths in fillets of red piranha, Pygocentrus
nattereri (Kner, 1858), with a note of a possible zoonotic potential of its consumption in nature. We
examined 40 fish caught in lakes in the floodplain of the Solimões River, located in the municipalities
of Coari and Manaus in central Amazonia, Brazil. All fish examined were adults and had yellow cysts
in the musculature. Of the 69 cysts found, 66 were identified as metacercariae of Clinostomum
marginatum (Braun, 1899) and 3 as Austrodiplostomum compactum (Lutz, 1928). The analysis shows
a higher susceptibility of P. nattereri for metacercariae of C. marginatum. The fact that metacercariae
were found in fillets of P. nattereri, suggests a risk situation, addresses a concern about its use in raw
form. It should not be consumed without proper inspection, processing and freezing, since ceviche, is
readily consumed, mainly in the border region of the upper and Western Amazonia.
Key Words: Brazil - Digenea - fish – metacercariae - Pygocentrus nattereri – Zoonoses.
El presente estudio investigó la presencia de helmintos en filetes de la piraña roja, Pygocentrus
nattereri (Kner, 1858), alertando sobre un posible potencial zoonótico de su consumo en la forma in
natural. Fueron examinados 40 peces capturados en lagos de varzea del Río Solimões, localizados
entre los Municipios de Coari y Manaus en la Amazonía Central, Brasil. Todos los peces analizados
fueron adultos y presentaban quiste amarillos en la musculatura del filete. Fueron encontrados 69
quistes e identificadas las metacercárias de las especie Clinostomum marginatum (Braun, 1899) (66
quistes) y Austrodiplostomum compactum (Lutz, 1928) (3 quistes). El análisis demuestra una mayor
susceptibilidad de P. nattereri para metacercárias de C. marginatum. El hecho de haber sido
encontradas metacercárias en filetes de P. nattereri, genera una situación de riesgo, remitiendo la
preocupación en cuanto a su consumo en la forma cruda, no debiendo ser consumido sin una adecuada
inspección, procesamiento y congelamiento, considerando que el cebiche es un alimento bastante
consumido, principalmente en la región fronteriza del Alto Solimões en la Amazonía Occidental.
Palabras claves: Brasil - Digenea- metacercarias- pez- Pygocentrus nattereri- Zoonosis.
MATERIAL AND METHODS
INTRODUCTION
Human cases of parasitism transmitted by fish
consumption have been reported in several
countries, especially in those where raw fish
dishes are part of their staple diet (Hirai et al.,
1987;Umegai et al.,1990; Barros et al., 2006;).
Human beings are also liable to get intoxicated
through the ingestion of dead anisakids larvae
(Nematoda) (Audicana et al., 2002), as well as
Clinostomidae digenetic trematode
metacercariae (Kifume et al., 2000), lying
within the fish's musculature.
Fish trematode zoonoses have great medical
importance in far eastern and southeastern
Asia, where up to 30 million people might be
infected. Infection comes through fresh water
fish, which makes up these people's main
source of protein. The ethological agents are
several Digenea species, which cause high
morbidity but are seldom the cause of death.
Transmission comes through the ingestion of
metacercariae lying mainly in fresh water fish
muscular and subcutaneous tissue (Santos,
1995; Ferre, 2001).
Most fish parasites which have been reported
occurring in man belong to Digenea. Among
them, Clonorchis sinensis (Looss,1907),
Diplostomum spathaveum (Rud, 1819),
Stellantchasmus falcatus Onji & Nishio, 1915,
Procerovum varium Onji & Nishio, 1916,
Nanophyetus schickhobalowi Skrjabin and
Podjapolskaja, 1931, Cryptocotyle lingua
(Creplin, 1825), Metorchis conjunctus
(Cobbold, 1860), Echinoschasmus perfoliatus
(Ratz, 1908), Echinostoma hortense,
Clinostomum complanatum Rudolphi, 1814,
Pseudamphistomum truncatum (Rudolphi,
1819) and Isoparorchis hypselobagri Billet,
1898 (Okumura et al., 1999).
Often infections are rare and occasional, as it
happens with Clinostomum marginatum
(Braun, 1899) and Austrodiplostomum
compactum (Lutz, 1928), and usually present
no pathogenic importance, except when
dealing with abundant infections. In the case of
A. compactum, human parasitose has been
associated to the condition known as parasitic
laryngopharangitis which, in its acute forms,
may lead to death through suffocation (Eiras,
1994).
Pygocentrus nattereri (Kner, 1958) commonly
known as red piranha or caju piranha, belongs
to Characiformes order, Characidae family and
is common in lentic environments (Santos et
al., 2006). It is a medium-sized fish, with a
standard length of nearly 25 cm. It occurs in the
Amazonas, Paraná-Paraguai and Essequibo
River basins, as well as in Brazil's northeastern
region Rivers (Jégu, 2003).
Its parasitic fauna is rather diversified, and it is
thought to be the Amazonian fish species
bearing the largest number of known parasite
species, with 35 of them having already been
reported as belonging to Monogenoidea,
Copepoda, Branchiura, Nematoda and Isopoda
Classes (Thatcher, 2006).
Pygocentrus nattereri is much consumed by
the local population which considers it as a
commercial fish (Santos et al., 2006). The lack
of sanitary inspection in order to detect
pathogens in its fillets favors the transmission
of zoonoses, mainly when they are ingested
while still raw, or not having been subjected to
proper boiling procedures (Barros et al., 2007).
The present study aims to investigate the
presence of helminths in P. nattereri fillets,
calling attention to their zoonose acquiring
potential if consumed in natura.
Fish captures, for parasitological analyses,
were undertaken from March to June and from
September to December 2008, which cover the
high and low water seasons in the central
Amazon. We determined the fishing effort
standard by using gill nets placed at random
i n t h e f o l l o w i n g f i v e l a k e s :
9
Morais et al.
Clinostomum and Austrodiplostomum on Pygocentrus
Lago do Baixio (S 03°17'27.2”/ W
60°04'29.6”) located at Iranduba Township.
Lago Iauara (S 03°36'39.2”/ W 61°16'33.0”)
sited at Manacapuru Municipality. Lago Ananá
(S 03°53'54.8”/ W 61°40'18.4”) located at
Anori County. Lago Araçá (S 03°46'15.8”/ W
62°20'10.3”) sited at Codajás Township. Lago
Maracá (S 03°50'32.8”/ W 62°34'32.4”)
localized at Coari Municipality. All of them on
a nearly 400 km long stretch on the Solimões
River floodplain between Coari County and
the Solimões and Negro Rivers merging area in
the Manaus neighborhood.
After capture the specimens of P. nattereri
were weighed, measured, and sex identified.
Afterwards, we removed the fillets and
immediately examined them for cysts in a light
box. We collected the cysts we found by hand,
broke them with the aid of thin dissecting
needles and observed them under light
microscope and stereoscope.
Digenea were cold fixed in AFA (alcohol,
formalin and acetic acid) with and without
compression. Specimens were stained with
Langeron's alcoholic carmine, dehydrated in
an ethyl alcohol series, cleared in beechwood
creosote and mounted in Canada balsam as
permanent slides. Parasitological indices, such
as, prevalence (P), intensity (I); infestation
mean intensity (IMI) and abundance (A) were
calculated and analyzed according to Bush et
al. (1997). Taxonomical identification is in
accordance with Travassos et al. (1969) and
Kohn et al. (1995).Vouchers specimens (PL
579, 580, 581 and 582) were deposited in the
colection of the Instituto Nacional de
P esquisas da Amazônia (INPA).
Forty P. nattereri were examined for
helminths. Mean standard weight and length
were 232.78± 91.92 g and 15.98± 1.87 cm,
respectively. All analyzed fish were adults and
bore yellow cysts in their fillet musculature
and were collected 69 metacercariae, as a
whole.
The cysts founded were morphologically
identified as metacercariae of A. compactum
and C. marginatum (Figure 1A,B). Sixty-six
metacercariae belong to C. marginatum and
only three were of A. compactum (Figure 2).
All sampled fish displayed higher
parasitological indices of C. marginatum than
of A. compactum (Table 1). From the analyzed
fish 37 (85%) were female and all of them
showed to be infested by C. marginatum,
whereas A. compactum was present only in
three male hosts (15%). We collected both
parasite species during low water season.
Parasitological indices of C. marginatum
infesting P. nattereri were: prevalence (P)
100%; intensity (I) 66 with the least of two or
the most of six metacercariae per fish,
infestation mean intensity (IMI) 3.36±1.56 and
mean abundance (MA) 3.36±1.56. A.
compactum parasitological indices were:
P=15%; (I)= 3(1-2); IMI= 1.57±0.71 and MA=
0.15±0.71.
RESULTS
DISCUSSION
Metacercariae of Clinostomidae use fish as
their intermediate hosts. The adult form infests
the mouth and esophagus of birds. Size and
color are their most significant trait. They are
the largest-sized metacercariae and bear bright
yellow coloring “yellow worms”. They occur
in all continents. The two most common
species are C. marginatum, which occurs
likewise in North, Central and South America,
and C. complanatum in Europe, Asia and
Africa (Thatcher, 1981; Bullard & Overstreet,
2008).
Three Amazonian fish species have already
been reported as being parasitized by
metacercariae of C. marginatum: Cichla
monoculus Spix and Agassiz, 1831 with
metacercariae encysted in the gills; Crenicihla
sp. in skin and fins (Thatcher, 1981) and
Semaprochilodus insignis (Jardine, 1841) in
the musculature (Castelo, 1984).
10
Neotrop. Helminthol., 5(1), 2011
Clinostomum marginatum is cited as parasite
mouth of several Amazonian piscivorous bird
species: Ardea cocoi Linnaeus, 1766; Jabiru
mycteria (Lichtenstein, 1819); Egretta
caerulea (Linnaeus, 1758); Phalacrocorax
brasilianus (Gmelin, 1789); Anhinga
(Linnaeus, 1766); Ardea cinerea Linnaeus,
1758; Ardea alba Linnaeus, 1758; Nycticorax
nycticorax (Linnaeus, 1758) and Butorides
striata (Linnaeus, 1758) (Travassos et al.,
1969).
Species of families Bolbophoridae,
Diplostomidae, Neodiplostomidae and
Strigeidae, members of the super family
Diplostomatoidea (Strigeoidea), were called
fish metacercariae in the old literature (Bullard
& Overstreet, 2008).
M a n y m e t a c e r c a r i a e o f t h e t y p e
“diplostomulum” accommodate themselves in
the eyes of their hosts, causing the formation of
cataracts and sometimes even their death. The
most important species presenting this
behavior is Diplostomum sphataceum
Rudolphi, 1819. The disease caused by
Diplostomum spp. was observed in more than
125 fish species with very wide geographical
distribution. Cercariae penetrate into the fish
and migrate to the eyes, probably through the
blood stream and parasitize mainly crystalline,
but may also go to the retina and vitreous
humor (Eiras, 1994).
Metacercariae of Strigeoidea were found
parasitizing various organs of Amazonian
fishes: Pterygoplichtys sp.; Astronotus
ocellatus (Agassiz, 1831); Oxydoras niger
(Valenciennes, 1821) and Geophagus
brasiliensis (Quoy & Gaimard, 1824)
(Thatcher, 1981). In Pterygoplichthys pardalis
(Castelnau, 1855) metacercariae of A.
compactum parasitized the eyes, stomach and
gonads (Porto, 2009).
In the present paper C. marginatum and A.
compactum are reported for the fist time
Clinostomum marginatum presented high
parasitological indices whereas just three
individual of A. compactum were collected.
Parasite species may occur in a distinct form in
male and female hosts, a characteristic which
has been frequently investigated in fish (Luque
et al., 1996; Poulin, 1996; Machado et al.,
2005). In the present paper C. marginatum only
parasitized P. nattereri females, while A.
compactum occurred only in males. The host's
sex had an influence on the parasite species,
pointing out there being ecological and
physiological factors determining this
specificity.
Parasites acquire more importance if or when
they are able to infecting human beings
(Castelo, 1984). Presently, there is more
emphasis on diseases originating from
contaminations by microbial organisms or
their toxins than on zoonoses or diseases
acquired by ingesting animals hosting these
parasites (Khamboonruang, 1991).
There are 17 confirmed cases of infections by
C. complanatum in Japan and one in Korea.
The main symptom is a queer feeling in the
throat a few days after ingesting any raw fish
dish. The treatment is carried out by an
otorrinolaringologist through the mechanical
removal of the metacercaria that lies fixed
either on the pharynges mucus wall or on the
arytenoids, being later identified (Furukawa &
Miyasato, 1987; Chung et al., 1995; Kifume et
al., 2000).
Finding metacercariae in P. nattereri fillets,
warns to there being a risky situation, and
stresses the care as to them being consumed
while still raw, making it clear they can become
a public health problem, once these fillets may
be consumed in the region as sushi, sashimi or
ceviche.
This paper brings an alert that P. nattereri
should only be consumed raw, after having
o
gone through prior freezing treatment at -20 C
11
Morais et al.
Clinostomum and Austrodiplostomum on Pygocentrus
for seven days or -30ºC for 15 hours,
corroborating with Barros et al. (2006), and
according to ANVISA technical ruling (2009)
for the consumption of raw or half boiled fish,
thus offering greater assurance to the
population.
ACKNOWLEDGE
Backing: FINEP, PETROBRAS (PIATAM
project), FAPEAM, INPA/MCT.
Table 1. Pygocentrus nattereri. parasitological indices, Mean fish weight (MW), fish mean standard
Length (MSL), prevalence (P), intensity (I), infection mean intensity (IMI) and mean abundance (MA). SE =
standard error.
12
Figure 1. Clinostomum marginatum (A) Bar = 200 μm. Austrodiplostomum compactum (B) Bar = 200 μm.
Clinostomun
marginatum
Austrodiplostomum
compactum
Host Sex
Neotrop. Helminthol., 5(1), 2011
BIBLIOGRAPHIC REFERENCES
Anvisa.2009.
Audicana, MT, Ansoetegui IJ, Torres, LF &
Kennedy, MW. 2002.
Barros, LA, Moraes-Filho, J & Oliveira, RL.
2006
Barros, LA, Moraes-Filho, J & Oliveira, RL.
2007.
Anvisa Alerta sobre o consumo
d e P e s c a d o C r u : a l e r t a e
recomendações referentes aos casos de
Difilobotríase no município de São
Paulo. Brasília, 6 de abril de 2005-
16:30h.<www.anvisa.gov.br/divulga/in
formes/2005/060405.htm>. Acesso em:
11 de dezembro de 2009.
Anisakis simplex:
dangerous - dead and alive? Trends in
Parasitology, vol.18, n.1, pp. 20-25.
. Nematóides com potencial
zoonótico em peixes com importância
econômica provenientes do rio Cuiabá.
Revista Brasileira de Ciência
Veterinária, vol. 13, pp. 55-57.
Larvas de nematóides de
importância zoonótica encontradas em
traíras (Hoplias malabaricus Bloch,
1794) no município de Santo Antonio do
Leverger, MT. Arquivos Brasileiros de
Medicina Veterinária e. Zootecnia,
vol.59, pp. 533-535.
Bullard, SA & Overstreet RM. 2008.
Bush, AO, Lafferty, KD, Lotz, MJ & Shotask,
AW. 1997
Castelo, FP. 1984.
Chung, DII, Moon, CW, Kong, HH, Choi, DH
& Lim, DK.1995.
E i r a s , J C . 1 9 9 4 .
Digeneans as enemies of fishes. In Fish
diseases. Eiras, J., H. Segner, T. Wahil,
and B. G. Kapoor. (eds.). Science
Publishers. Enfield, New Hampshire, pp.
817–976.
. Parasitology meets ecology
on its own terms. Journal of Parasitology,
vol. 83, pp. 575-583.
Acta Amazonica,
vol. 13, pp. 325-326.
The first human case
o f Cl inos tomu m c omp lana tum
(Trematoda: Clinostomidae) infection in
Korea. The Korean Journal of
Parasitology, vol. 33, pp. 219-223.
E l e m e n t o s d e
ictioparasitologia. Porto: Fundação
Eng. António de Almeida. 159 p.
Oc o r r ê n c i a de
Clinostomum marginatum Rudolphi,
1819 "Yellow-Spot Disease" em filé de
jaraqui (Semaprochilodus insignis
Shomburgk, 1814).
13
Figure 2. Metacercarie cyst standing out in Pygocentrus nattereri musculature fillet (C). Bar = 3 cm.
Morais et al.
Clinostomum and Austrodiplostomum on Pygocentrus
Ferre, I. 2001.
Furukawa, T & Miyazato, T. 1987.
Hirai, H, Olso, H, Kifune, T, Kiota, T &
Sakaguchi, Y. 1987.
Jégu, M. 2003.
Khambooruang, E. 1991.
Kifume, T, Ogata, M & Miyahara, M. 2000.
Kohn, A, Fernandes, BMM & Baptista-Farias,
MFD. 1995.
Anisakiosis y otras zoonosis
parasitarias transmitidas por consumo
de pescado. Revista Aquatic, vol.14, pp.
1-20.
A case of
Clinostomum sp. infection in human
pharynx. Medicine Journal Kinki
University, vol. 12, pp. 665-669.
Clinostomum
cornplanaturn infection in posterior wall
of the pharynx of a human. Japanese
Journal of Parasitology, vol. 36, pp. 142-
144.
Subfamily Serrasalminae. In:
Reis, RE, Kullander, SO, Ferraris, C.J.
(Eds). Check list of the freshwater fishes
of south and Central America.
(CLOFFSCA). EDIPUCRS, Porto
Alegre. pp. 182-196.
On emerging
problems in food-bome parasitic
zoonosis: impact on agricultores and
public health. Southeast Asian Journal
Tropical Medicine and Public Health,
vol. 22, pp.1-7.
The first case of human infection
w i t h Cl inos t omum ( Tre m a t o d a :
C l i n o s t o m i d a e ) i n Ya m a g u c h i
prefecture. Japan Medicine Bulletin
Fukuoka University, vol. 27, pp. 101-
105.
Metacerc a r i a e of
Diplostomum (Austrodiplostomum)
compactum (Trematoda,Diplostomidae)
i n t h e e y e s o f P l a g i o s c i o n
squamosissimus (Teleostei, Sciaenidae)
from the reservoir of the Hydroelectric
Power Sation of Itaipu, Brazil.
Memórias do Instuto Oswaldo Cruz, vol.
90, pp. 341-344.
Luque, JL, Amato, JFR & Takemoto, RM.
1996.
Machado, PM, Takemoto, RM & Pavanelli,
G C . 2 0 0 5 .
Okumura, MPM, Pérez, ACA & Espíndola, A.
1999.
Porto, DB. 2009.
Poulin, R. 1996.
Santos, GM, Efrem, JGF & Zuanom, JAS.
2006.
Santos, EAL. 1995.
Comparative analysis of the
communities of metazoan parasites of
Orthopristis ruber and Haemulon
s t e i n d a c h n e r i ( O s t e i c h t h y e s :
Haemulidae) from southeastern
Brazilian littoral: I. Structure and
influence of size and sex of hosts. Revista
Brasileira de Biologia, vol. 56, pp. 279-
192
D i p l o s t o m u m
(Austrodiplostomum) compactum (Lutz,
1928) (Platyhelminthes, Digenea)
metacercariae in fish from the floodplain
of the upper Paraná river, Brazil.
Parasitology Research, vol. 97, pp. 436 –
444.
Principais zoonoses parasitárias
transmitidas por pescado – revisão.
Re v ista Ed u caçã o C onti n uada
CRMVSP, vol. 2, pp. 66-80.
Metazoários parasitas de
Pterygoplichthys pardalis (Castelnau,
1855) (Ostheichthyes, Loricariidae) de
lagos de várzea do rio Solimões entre
Manaus e Coari, Estado do Amazonas,
Brasil. Dissertação de mestrado, INPA,
pp. 37.
Sexual inequalities in
helminth infections: a cost of being a
male? The American Naturalist, vol.
147, pp. 287-295.
ed
Peixes comerciais de Manaus. 1 .
Manaus:IBAMA/AM, ProVárzea, pp.
49.
Prevention and contrai of
food bome trematódas in cultured fish.
Infofish International Journal, vol. 2, pp.
57-62.
.
14
Neotrop. Helminthol., 5(1), 2011
Thatcher, VE. 1981.
Thatcher, VE. 2006.
Travassos, L, Freitas, JFT & Khon, A. 1969.
Umegai, T, Shint, T, Oda, M, Kifune, T &
Moci, M. 1990.
Patologia de peixes da
Amazônia brasileira, 1. Aspectos gerais.
Acta Amazonica, vol. 11 , pp. 125-140.
th
Amazon fish parasites. 2
Ed., Ed. Aquatic Biodiversity in Latin
America, Sofia, Moscow, 487 p.
Trematódeos do Brasil. Memorias do
Instituto Oswaldo Cruz, vol. 67, pp. 395-
399.
A case of acute laryngitis
caused by Clinostomum complanatum
with a complaint of throat irritation (in
Japanese). Jibi To Hinsho, vol.36, pp.
665-668.
Author for correspondence /Autor para
Aprigio Mota Morais
INPA- Instituto Nacional de Pesquisas da Amazônia,
Laboratório de Parasitologia e Patologia de Peixes,
Manaus, AM. Brazil.
correspondencia:
15
Received January 6, 2011.
Accepted March 15, 2011.
Morais et al.
Clinostomum and Austrodiplostomum on Pygocentrus
E-mail/correo electrónico:
aprigiomota@yahoo.com.br.
