Melanoides tuberculata (Mollusca: Thiaridae) as an intermediate host of Centrocestus formosanus (Trematoda: Heterophyidae) in Brazil

Melanoides tuberculata (Mollusca: Thiaridae) como hospedeiro intermediário de Centrocestus formosanus (Trematoda: Heterophyidae) no Brasil

PARASITOLOGY

Melanoides tuberculata (Mollusca: Thiaridae) as an intermediate host of Centrocestus formosanus (Trematoda: Heterophyidae) in Brazil

Melanoides tuberculata (Mollusca: Thiaridae) como hospedeiro intermediário de Centrocestus formosanus (Trematoda: Heterophyidae) no Brasil

Hudson Alves Pinto; Alan Lane de Melo

Laboratório de Taxonomia e Biologia de Invertebrados, Departamento de Parasitologia, Instituto de Ciências Biológicas, UFMG, Belo Horizonte, MG, Brasil

^{Correspondence to}

SUMMARY

Pleurolophocercous cercariae emerged from naturally infected Melanoides tuberculata from Minas Gerais State, Brazil, were used to perform experimental infection of laboratory-reared Poecilia reticulata. Mature metacercariae were obtained from the gills of fishes and force-fed to Mus musculus. The adult parasites which recovered from small intestines of mice were identified as Centrocestus formosanus. This is the first report of M. tuberculata as intermediate host of this heterophyid in Brazil.

Keywords:Centrocestus formosanus; Melanoides tuberculata; Brazil; Experimental infection; Trematoda.

RESUMO

Cercárias do tipo pleurolofocerca emergidas de Melanoides tuberculata naturalmente infectados coletados na represa da Pampulha, Belo Horizonte, Minas Gerais, Brasil, foram utilizadas para a infecção experimental de Poecilia reticulata criados em laboratório. Metacercárias maduras foram obtidas nas brânquias dos peixes e administradas por via oral a Mus musculus. Parasitos adultos recuperados no intestino delgado dos camundongos foram identificados como Centrocestus formosanus. Este é o primeiro relato de M. tuberculata como hospedeiro intermediário deste parasito no Brasil.

Melanoides tuberculata (Müller, 1774), an exotic mollusc introduced in Brazil in the late 1960s⁴⁰, has been found in several states in Brazil¹². Despite some studies related to the interaction between M. tuberculata and planorbid molluscs which transmit Schistosoma mansoni Sambon, 1907 in the country^13,14, the possible damage on the native fauna as alien species and their potential to act as parasite vector in the country have not yet been evaluated. In this later aspect, some studies had already reported the finding of M. tuberculata harboring pleurolophocercous cercariae in Brazil^5,6,39, but these larvae, characterized as Pleurolophocercous type, still remain without a specific identification.

In the present study, malacological surveys were performed at Pampulha dam (an artificial lake located between 43⁰ 58' and 44⁰ 01'W and 19⁰ 50' and 19⁰ 52'S, in the northern region of the city of Belo Horizonte and 2 km away from the campus of "Universidade Federal de Minas Gerais") from March 2006 to July 2009. Molluscs were collected with a scoop net and long forceps. After a preliminary trial, molluscs were placed in a thermal box, labeled and shipped to the laboratory. Then they were placed individually in plastic recipients containing 5 mL of tap water and left overnight at room temperature. Thiarids were examined with a stereomicroscope before and after artificial photostimulation for two hours. Emerged larvae were collected with micropipette, transferred to glass slides and analyzed with vital stains (0.05% neutral red, 0.05% toluidine blue, 0.05% alizarin red) or fixed and then stained with acetic carmine and mounted in Canada balsam²³.

Cercariae were concentrated by filtration using a Buchner funnel³¹ and one hundred larvae in 5 mL were used for individual infection of 30 adult specimens (2-3 cm of length) of laboratory reared Poecilia reticulata Peters, 1859 (Pisces: Poeciliidae). Thirty days after infection the surviving fishes were necropsied in order to recover metacercariae. Cysts obtained from gills of experimentally infected fish were administered per os to five mice of the AKR/J strain. Fifteen days after infection, mice were sacrificed by cervical dislocation according to the local animal experimentation ethics committee (CETEA/UFMG), and their intestines were thoroughly and longitudinally opened in Petri dishes containing 0.85% saline solution and adult parasites were recovered from the proximal third of the small intestine. Parasites were studied alive or then placed between the slide and the coverslip with slight pressure, fixed in cold 10% formalin, stained with acetic carmine, dehydrated, cleared in benchwood and mounted in Canada balsam.

Drawings were made using a camera lucida and specimens were measured with a millimitered eyepiece. Photographic documentation was performed in light microscope. The results were compared with taxonomical keys⁷ and morphological descriptions from several authors^{3,9,10,17,18,22,29}. The specimens studied were deposited at the collection of Department of Parasitology (DPIC), UFMG.

From 3,834 specimens of M. tuberculata collected, 268 (7.0%) were naturally infected by larval trematodes preliminarily characterized as belonging to the group of Pleurolophocercous cercariae. These larvae encysted in P. reticulata and out of thirty fish experimentally infected, twenty survivors were analyzed, all of them presenting viable metacercariae which were used for inoculation of mice. Fifty ovigerous worms with 32 circumoral spines were obtained from the experimentally infected mice and identified as members of family Heterophyidae Leiper, 1909, subfamily Centrocestinae Looss, 1899 and the species Centrocestus formosanus (Nishigori, 1924) (Fig. 1). The morphology and morphometry of the developmental stages obtained are in accordance with the original description and with experimental studies conducted by different authors (Table 1). Measurements are presented by the mean followed by the range in parenthesis.

C. formosanus was described in 1924 from adult parasites recovered from natural and experimental definitive hosts in Taiwan²⁹. The life cycle of this trematode was also studied in China, where M. tuberculata were recognized as the first intermediate hosts and fishes and frogs as second intermediate hosts^9,10. The species was introduced in the New World in the late 1950s²², possibly by thiarids, migratory birds and/or exotic fishes naturally infected.

Among fish, several species of different families have already been reported as naturally and experimentally infected in many countries. The development of metacercariae on gills causes pathological changes which, in some cases, as high parasite burden, may cause the death of these hosts and damage in aquaculture^{2,15,24,25,26,28,30,35,36,42}.

Adult parasites, in turn, have already been reported naturally in fish-eating birds (Butorides striatus (Linnaeus, 1758); Bubulcus ibis coromandus (Boddaert, 1783) and Nycticorax nycticorax (Linnaeus, 1758)), and mammals, including human beings^{9,11,19,20,21,28,29,32,44}.

Several experimental models, among birds (chickens, ducks, pigeons) and mammals (dogs, cats, Rhesus monkeys, mice, hamsters, rats and rabbits), were found to be susceptible to infection with the trematode in several parts of the world^{3,9,18,22,27,28,34,38}, being considered a parasite with low intraspecific variability among different hosts and places³⁶.

In the American continent there are some published reports of the parasite in the USA^22,24,25,26, Mexico^1,3,36, Venezuela¹⁸, Colombia^41,43. Recently the occurrence of C. formosanus in Poecilia vivipara (Bloch & Schneider, 1801), from Rio de Janeiro, Brazil was reported in a congress communication³⁷. However, until the present study the intermediate host of this heterophid was unknown in this country.

In Minas Gerais State, M. tuberculata was introduced in the 1980's⁸ reaching a high population density one decade after the first report⁴ and today is considered to be a well established population. Due to the existence of several species of piscivorous birds, resident and visiting ones³³, as well as a rich ichthyofauna¹⁶, the Pampulha dam has become an appropriate environment to maintain the life cycle of C. formosanus.

The control strategy against the spread of this trematode and the intermediate host, M. tuberculata, need to be implemented to prevent the spread of human and veterinary cases of this parasite in Brazil. This is the first report of C. formosanus in M. tuberculata from Brazil and of this Heterophyidae in the state of Minas Gerais.

ACKNOWLEGMENTS

To Mr. Airton Lobo for technical assistance and to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for financial support and scholarship to H.A.P.

Received: 22 February 2010

Accepted: 1 June 2010

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Publication Dates

• Publication in this collection
  25 Aug 2010
• Date of issue
  Aug 2010

History

• Received
  22 Feb 2010
• Accepted
  01 June 2010