Bull. Eur. Ass. Fish Pathol., 35(2) 2015, 41
Parasites and pathological condition ȱ ȱȱȱȱȱ Cassino beach, southern Brazil
Y. B. M. Carvalho, L. H. da Silva Poersch and L. A. Romano*
Laboratório de Imunologia e Patologia de Organismos Aquáticos, Estação Marinha de Aquicultura, ȱȱęǰȱȱȱȱȱ ǰȱȱşŜŘŖŗȬşŖŖǰȱǯ
Abstract ȱȱ¢ȱ ȱȱ ȱ ȱȱǰȱAmiantis purpurata (Veneridae) and Mactra isabelleana ǻǼȱȱȱǰȱȱ ȱȱȱǰȱȱ£ǯȱ ȱȱȱřŖȱȱȱȱȱ ȱ¢ȱȱȱȱȱȱȱȱȱ ȱȱȱȱȱ¢ȱȱȱȱǯȱȱȱȱȱ A. purpurataȱ DZȱ¢ȱȱȱNematopsis-like gregarine (Apicomplexa) (prevalence = 20%) in ȱ¢ȱȱ ȱȱDzȱȱȱ ȱ¢ȱȱȱȬȱ ȱ ȱȱȱȱȱȱȱȱǻȱƽȱŞřǯśƖǼǰȱȱȱȱ ȱȱȱDzȱ¢ȱȱȱȬȱǼȱȱȱȱǻȱƽȱŗřǯřƖǼȱ ¢ȱȱȱȱDzȱȱęȱȱǻǼȱ¢ȱȱ the gills (prevalence = 3.3%) without host reaction. The parasites recorded in M. isabelleana were: Pseudoklossia-like coccidians (Apicomplexa) in the kidney (prevalence = 20%) hypertrophying the ȱȱ ȱȱȱǰȱȬȱȱ¢ȱęȱȱȱ ǻȱƽȱřǯřƖǼǯȱȱȱȱęȱȱȱȱȱM. isabelleana and the second with A. purpurata ǻȱęȱȱȱ£ȱǼǯ
Introduction It is well known that diseases can dramatically Recently, histopathological studies were per- ěȱȱȱȱȱȱȱȱ ȱ ȱȱ¢ ȱȱMesodesma mac- and aquaculture production systems (Lauck- troidesȱȱȱȱǰȱȱ ȱȱ ǰȱŗşŞřDzȱȱȱǯǰȱŘŖŗŚǼǯȱȱǰȱȱ Sul State, southern Brazil (Carvalho et al., 2013a; ȱȱȱȱȱȱ£ȱȱ Carvalho et al., 2013b). The Cassino Beach is scarce. Investigations conducted in the north- ȱȱȱȱȱȱȱȱ ȱ ȱȱȱȱ¢ǰȱȱȱȱȱ (approximately 220 km) (Seelinger et al., 1997) Bahia (Zeidan et al., 2012), Paraíba (da Silva et al., where the subtidal clam Mactra isabelleana ǻȂ- 2013), Ceará (Sabry et al., 2007; Sabry et al., 2013), ¢ǰȱŗŞŚŜDzȱǼȱȱȱȱȱ and also in the south in Santa Catarina State (da Amiantis purpurataȱǻǰȱŗŞŗŞDzȱǼȱ Silva et al., 2002; da Silva et al., 2012), where (Rios, 2009) occur. mussels Perna perna, oysters Crassostrea gigas and the scallop Nodipecten nodosus ȱǯ The purple clam, a warm-temperate species,
ȘȱȱȂȱȬDZȱȓ¢ǯ 42, Bull. Eur. Ass. Fish Pathol., 35(2) 2015
ȱ ȱȱȱĴȱȱ ¢ȱȱǰȱȱȱȱȱ ȱȱȱȱȱȱȱ- ȱ ȱȱȱȱȱȱȱ ÇȱȱǻŘŖǚǰȱ£ǼȱȱȱÇȱ ȱ ę¡ȱȱȂȱǯȱ ȱȱ¢ȱ ǻŚŗǚǰȱǼȱǻǰȱŘŖŖşǼǯȱȱȱȱȱ the samples including gills, digestive gland, Ȭ ȱ ȱȱ¡ȱ¢ȱȱŚŘȱ kidney and mantle, were dehydrated in an ® years and this species is harvested, recreationally ethanol series, embedded in Paraplast , sec- and commercially along the Argentinean coast tioned (5 μm thick) and stained with haema- (Morsan and Orensanz, 2004). The mactrid clam toxylin and eosin (H&E). The histological slides M. isabelleana ȱȱ¢ȱĴǰȱȱ were observed with an optic microscopy in ȱŘśȱȱȱȱȱȱ ȱǻ£Ǽȱȱ ȱȱȱȱȱȱȱ ȱȱȱÇȱǻǼȱǻǰȱŘŖŖşǼǯ alterations.
ȱȱȱ¢ȱȱȱȱȱ Results ȱȱȱȱȱȱȱȱȱ Parasites and pathological signs in the purple ȱȱȱ¢ȱȱ ȱȱȱ clam Amiantis purpurata ǯȱȱȱȱȱȱȱ ¢ȱȱȱNematopsis-like gregarine (Api- ȱȱȱǰȱȱęȱȱȱȱ complexa: Eugregarinidae) were recorded in ȱǯȱǰȱ¢ȱȱȱęȱ ŘŖƖȱȱȱȱȱ¢£ǯȱȱ¡ȱ resource in the region. ȱŞȱ¢Ȧȱȱ ȱǯȱ These parasites were recorded in the Leydig Several studies demonstrated that stranded sub- tissue eliciting no evidence haemocytic response tidal clams washed up to the beach are sick and by the hosts (Figure 1). ǰȱȱȱ ȱĜ¢ȱȱȱ sediment (Bowers, 1969; Cremonte et al., 2001; Sporocysts containing cercariae and germ balls ǰȱŗşŞřǼǯȱǰȱȱȱȱȱȱ ǻǰȱǯǯȱǼǰȱ¢ȱ- is to describe the parasites and pathological ing the digestive gland (Figure 2), gonad, and ȱȱȱȱȱ ȱȱ kidney were recorded with high prevalence (A. purpurata and M. isabelleana) collected at ǻŞřǯřƖǼǯȱ ȱȱ£ȱǰȱȱ ȱȱ ȱȱȱȱȱȱȱȱǯ possible to see gametogenesis, while in the ȱȬȱȱȱ ȱȱȱ Material and methods ȱȱȱȱȱȱȱȱ ¢ȱȱȱȱȱ ȱȱ gametogenesis. ¢ȱȱȱȱȱ£ȱȱȱȱ ȱǻřŘɃŗşȂȱȱśŘɃŗŝȂȱǼǯȱM. isabelleana (shell ¢ȱȱȱȱȱ height = 41 ± 3.32 mm) and A. purpurata (shell ȱȱȱǻǯǯȱ¢Ǽȱȱ- height = 35.22 ± 4.74 mm) were collected in ȱȱȱȱ ȱȱ ŘŗȦŖşȦŘŖŗŗȱǻȱǰȱ ȱȱ only in the gills (prevalence = 13.3%). The target ƽȱŗŚǚǰȱ¢ȱƽȱřŖƗǼȱȱřŖȦŖŞȦŘŖŗřȱǻȱ ȱȱȱȱ ȱȱȱ ǰȱ ȱȱƽȱŗŚǚǰȱ¢ȱƽȱ ȱȱȱȱǯȱȱ¢ȱȱ ŘŜƗǼǰȱ¢ǯ ȱ ȱ¢ȱȱȱȱȱȱȱ Bull. Eur. Ass. Fish Pathol., 35(2) 2015, 43
Figure 1.ȱ¢ȱȱNematopsisȬȱȱǻ¡Ǽȱȱȱ¢ȱȱȱȱȱȱ Amiantis purpurata. Note the worm-like sporozoite densely eosinophilic with a basophilic nucleus enclosed by a thick hyaline capsule (arrows). H&E- staining, Bar = 25μm.
Figure 2.ȱ¢ȱȱȱǻǼȱȱěȱȱȱȱǻ Ǽȱȱȱ ȱȱȱȱȱȱAmiantis purpurata. H&E - staining, Bar = 50μm. 44, Bull. Eur. Ass. Fish Pathol., 35(2) 2015
£ȱȱ ȱ¢ȱęȱȱ ȱ ȱȱ ȱȱȱȱ¢£ȱȱ sporocysts (Figure 3). ȱȱ¢ȱǻ¢ȱȱȱȱǼȱ it was possible to observe several parasites and In one A. purpurata (prevalence = 3.33%), two pathological signs in the histological slides. The ęȱ£ȱ ȱȱ ȱȱ ¢ȱȱȱA. purpurata and M. isabellea- ȱȱȱȱȱ ȱȱȱ naȱȱ¢ȱȱȱȱȱǰȱ Ě¢ȱȱȱȱȱǻȱŚǼǯ ȱȱȱȱ¢ȱȱȱ in the intertidal zone. Parasites and pathological signs in Mactra isabelleana According to Cremonte (2011), Digenea in the ęȱPseudoklossia-like coccidians (Api- larval stage are the most important metazoan complexa) were recorded in the epithelial cells ȱȱȱǯȱȱ- ȱȱ¢ȱȱŘŖƖȱȱȱȱ¢£ǯȱȱ netic trematode uses gonad tissue in the bivalve ¡ȱ¢ȱȱ ȱŜȱȦȱ ęȱȱȱǻ¢ȱǼȱ¢ȱ histological slide. The parasitized cells were may cause castration. On the Brazilian coast, hypertrophied or ruptured and no host reaction ȱǰȱ¢ȱȱȱȱȱ was recorded (Figure 5). Bucephalus sp. in the mussel Perna pernaȱȱ ȱȱȱȱȱ- ¢ȱǻȱǯǯȱ¢Ǽȱ- ȱȱȱȱȱȱȱȱǻȱ taining germ balls and cercariae were observed Silva et al., 2002). ¢ȱȱȱȱȱ ȱM. isabelleana (prevalence = 6.6%). The interlamellar connec- ȱȱȱȱȱę¢ȱȱ ȱȱȱȱȱ ȱ¢ȱęȱ ȱ digeneans have to their primary host, the spo- sporocysts (Figure 6). ¢ȱȱȱȱȱȱȱȱ the purple clam observed in the present study Discussion are probably the same parasite described by According to Bower et al. (1994), each molluscan Cremonte et al. (2001) as Monorchiid cercaria sp. ȱȱȱ¢ȱȱȱ¢ȱȱ Cremonte et al. (2001) reported high prevalence organisms, comprising viruses, prokaryotes, ǻşŝƖǼȱȱMonorchiid cercaria sp. in purple clams protozoans and metazoans parasites, although ȱȱȱȱȱȱȱȱȱ ¢ȱ ȱȱȱȱȱȱȱ in the Argentinean coast. They suggest that disease. these parasites debilitates the hosts impairing their ability to burrow in the sand, and also ȱȱ¢ȱ ȱȱěȱȱ ȱȱȱȱȱǰȱȱȱ similarities in the parasitic community between monorchiid occupies almost all the haemocoele Amiantis purpurata and Mactra isabelleana, which ȱȱȱȱȱǰȱȱȱȱ occupied the same habitat in the subtidal region observed situation in the present study. ȱȱǰȱȱA. purpurata is probably longer-lived (up to 40 years) than M. isabelleana ȱȱȱȱȱȱȱ¢£ȱȱ ǻȱȱȱȱ¢ȱȱM. isabelleana). the present study was represented by a debil- Bull. Eur. Ass. Fish Pathol., 35(2) 2015, 45
Figure 3.ȱ ȱȱȱȱȱAmiantis purpurata ¢ȱȱ¢ȱ¢ȱȱȱ ǻǰȱ¢ȱ¢Ǽȱǻ Ǽǯȱ ǭȱȬȱǰȱȱƽȱřŖŖȱΐǯȱ DZȱ ȱęȱȱ ȱȱ¢ȱ¢ȱ ȱȱǻ ǼǯȱȱƽȱśŖȱΐǯ
Figure 4.ȱęȱ£ȱǻǼȱ¢ȱȱȱȱȱȱȱ ȱ Amiantis purpurata (arrows). H&E - staining, Bar = 50 μm. 46, Bull. Eur. Ass. Fish Pathol., 35(2) 2015
Figure 5.ȱ ¢ȱȱMactra isabelleanaȱȱ¢ȱȱPseudoklossia-like in the microgamont stage. Note ȱȱ¢ȱǻ Ǽǯȱ ǭȱȬȱǰȱȱƽȱŘśȱΐǯ
Figure 6.ȱ ȱȱMactra isabelleana ¢ȱȱ ȱ¢ȱǻȱǯǯȱ¢Ǽȱǻ Ǽǯȱ H&E - staining, Bar = 100μm. Bull. Eur. Ass. Fish Pathol., 35(2) 2015, 47 itated group that was washed up to the beach ȱȱȱ ȱȱȱǰȱ ȱȱǰȱȱȱȱȱȱȱȱ ȱȱ¢ȱ¢ȱȱȱ ȱ¢ȱ¢ȱǯǯȱMonorchiid cercaria ȱȱȱȱȱȱěȱ in a “healthy” sample collected in the subtidal on the host physiology (Sindermann, 1990). zone should not be as high as reported in the ȱȱ£ȱǰȱ¢ȱȱȱȱ present study. were recorded in several regions and bivalves. For example, Nematopsis were detected in the ȱȱȱȱȱȱȱȱȱ mytilid Mytella guyanensisȱȱ£ȱ ȱȱȱȦȱȱ ȱ- (northern Brazil) (Azevedo and Matos, 1999); served only in A. purpurata in high prevalence in M. guyanensis and venerid clam Anomalocar- ǻŞśƖǼȱȱ¢ǯȱȱȱȱǰȱ- dia brasiliana ȱȱǻȱ£Ǽȱ ¢ȱ¢ȱęȱȱȱȱ ȱ- (Boehs et al., 2010) and in the mangrove oyster corded in low prevalence in both clams species Crassostrea rhizophoraeȱȱȱȱȱ ¢£ȱȱȱȱ¢ǯȱȱ¡¢ȱȱ (southern Brazil) (Sabry et al., 2011). these parasites is still unknown; however they ¢ȱȱȱȱ¢ȱ¢ǰȱ Coccidians in the genus Pseudoklossia are ȱȱȱȱȱǰȱȱǰȱ ȱȱȱȱȱȱ ǯȱ as reported by Gilardoni et al. (2011). ȱȱ¢ȱȱȱȱ¢ȱȱ ȱȱ ȱ¢ȱȱ¢¢ȱȱ Gilardoni et al. (2011) recently described an apo- the renal cells (Carballal et al., 2001; Lauck- ¢ȱȱȱȱȱȱA. pur- ǰȱŗşŞřǼǯȱȱȱ ǰȱȱȱȱȱ purataȱȱȱȱǯȱȱȱ ȱȱȱęȱȱ- ȱǰȱȱȱȱȱȱȱ asitizing the mangrove oyster C. rhizophorae ȱ¢ȱȱȱ ȱȱȱȱ ȱȱȱǻȱ£Ǽȱ occurrence are recorded in the world. (Azevedo et al., 2005). So, the present study is ȱȱȱȱȱȱȱ The other parasites recorded in the present ȱ£ǯȱ study (Pseudoklossia-like coccidians, Nematop- sisȬȱǰȱȱęȱ£Ǽȱ Acknowledgements were reported in numbers too low, or without YBM Carvalho was supported by scholarship ȱ¢ǰȱȱȱȱȱ ȱȃȱȱȱȱ impacts on the clam populations. According to ÇęȱȱàȄȱǻǼȱȱ£- Carballal et al. (2001), Pseudoklossia and Nema- ian Government. Luis A. Romano and Luis H. topsis induce light to moderate damage to the ȱȱ¢ȱȱ - cockles Cerastoderma eduleȱȱ ǰȱǰȱ ȱȱǯ ȱȱȱ¢ȱȱǯȱ References Gregarines in the genus Nematopsis (Apicom- Azevedo C and Matos E (1999). Description ȱNematopsis mytella sp. (Apicomplexa), plexa) are sporozoans that use marine bivalves ȱȱȱȱMytella guyanensis ȱȱȱǻǰȱŗşŞřǼǯȱ- ŚŞǰȱǯȱǯȱǯȱȱǯǰȱřśǻŘǼȱŘŖŗś
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