RESUMEN

The mangrove oyster, Crassostrea rhizophorae (Bivalvia, Ostreidae) is commonly collected by fisherwomen in the estuaries of the Ceará State (CE), Northeastern Brazil. Despite the socioeconomic importance of this natural resource, there are few studies on the health of the oysters in this region. This study aimed to survey pathological changes in the mangrove oyster C. rhizophorae in the estuary of the Pacoti River, CE. Adult oysters were collected in August 2008 (N=450) and December 2009 (N=450) at three sites of the Pacoti estuary and in 2010 (N=600) samplings were done quarterly at one site which has showed the higher prevalence de Perkinsus. Macroscopical and histological analyses were used to evaluate pathological changes, Ray's Fluid Thioglycollate Medium (RFTM) to detect Perkinsus spp. and polymerase chain reactions (PCR) and DNA sequencing to identify Perkinsus species. In 2009, RFTM assay detected Perkinsus sp. infecting the tissues of C. rhizophorae with low prevalences of 1.3%, 6.7% e 7.3% in sites 1, 2 and 3, respectively, and in 2010, in site 3, prevalence was 2% (12 of 600 oysters). PCR did not confirm any positive case in 2009 and only 5 in 2010. The phylogenetic analyses strongly indicate that the Perkinsus species infecting oysters C. rhizophorae of this study belongs to Perkinsus beihaiensis. The histology confirmed 11 cases of Perkinsus sp. infecting the C. rhizophorae in 2009, and only two cases in 2010. Nematopsis sp. was the protozoan observed with greater prevalence (up 96.7%). Other found protozoa were: Trichodina, Sphenophrya, Ancistrocoma - like and an unknown ovarian parasite. The metazoa found were the polychaete Polydora with high prevalences, a turbellarian, possibly of the genus Urastoma, an unidentified digenean metacercariae and larvae of cestode Tylocephalum. A continuous monitoring of diseases in bivalves from this natural population is recommended, since the phylogenetic analyses indicate the occurrence of P. beihaiensis infecting oysters C. rhizophorae whose pathogenic potential is unknown.

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The orange disease is considered the main parasitosis in Perna perna mussel. It is caused by a complex life cycle bucephalid, involving three hosts, among them mollusks and fishes. With the aim of contributing to the knowledge of orange disease in mussel culture, the parasite life cycle was investigated. Experimental studies and monthly samples in the study area allowed the identification and characterization of the Bucephalus margaritae life cycle. Larvae and adults of B. margaritae were fixed in 5% formaldehyde, stained with Gomori’s trichrome, clarified in creosote and mounted in Canada balsam. The cercariae are found in the first intermediate host P. perna inside the sporocysts, which have the form of orange and ramified filaments. The metacercariae encysts in the gills and gill cavity of the second intermediate host, the blenniid Hypleurochilus fissicornis. The definitive host Menticirrhus americanus is infected ingesting blenniids parasitized with metacercariae. The high parasitological indexes of B. margaritae suggests that M. americanus and H. fissicornis act as the main definitive and intermediate hosts, respectively, in the trematode life cycle. The blenniid H. fissicornis is a new intermediate host to the species. Key words: Bucephalus margaritae, Hypleurochilus fissicornis, life cycle, orange disease, Perna perna.

A bucefalose é considerada a principal parasitose do mexilhão Perna perna. É causada por um Bucephalidae, de ciclo de vida complexo, envolvendo três hospedeiros, entre eles, moluscos e peixes. Com o objetivo de se contribuir para o conhecimento da bucefalose em mexilhões de cultivo, foi investigado o ciclo de vida deste parasito. Estudos experimentais e coletas mensais na área de estudo permitiram caracterizar o ciclo de vida de Bucephalus margaritae. Larvas e adultos do parasito foram fixados em formol 5%, corados com tricrômico de Gômori, clarificados em creosoto e montados em lâminas permanentes com bálsamo do Canadá. As cercárias ocorrem no primeiro hospedeiro intermediário P. perna, no interior dos esporocistos que têm a forma de filamentos ramificados e alaranjados. As metacercárias encistam nas brânquias e cavidade branquial do segundo hospedeiro intermediário, o blenídeo Hypleurochilus fissicornis. O hospedeiro definitivo, Menticirrhus americanus, é infectado quando ingere blenídeos com metacercárias. Os elevados índices parasitológicos de B. margaritae sugerem que M. americanus e H. fissicornis sejam os principais hospedeiros definitivo e intermediário, respectivamente, deste trematódeo. O blenídeo H. fissicornis é um novo hospedeiro intermediário para a espécie.

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Protozoan parasites of the genus Perkinsus are considered important pathogens responsible for mass mortalities in several mollusk species worldwide. In the present study we describe for the first time a parasite of the genus Perkinsus infecting the mangrove oyster Crassostrea rhizophorae from the Brazilian coast. Prevalence of this parasite was low in the Pacoti River estuary (Ceará, northeast Brazil) and absent in oysters from southern Brazil. Oyster gill and rectum tissues incubated in Ray's fluid thioglycollate medium (RFTM) revealed the presence of spherical hypnospores (5 to 55 microm diam.). Histological analysis showed the occurrence of typical signet-ring trophozoites and schizonts (3 to 6 microm diam.) infecting connective tissues of several organs and digestive epithelia. PCR assays specific to the genus Perkinsus, followed by cloning and sequencing of the internal transcribed spacer (ITS) region of the ribosomal ribonucleic acid (rRNA) gene complex, confirmed a close phylogenetic relationship between Brazilian Perkinsus sp. and P. beihaiensis infecting Chinese oysters.

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