ABSTRACT
Ostreopsis cf. ovata is a benthic dinoflagellate that produces palytoxin-like compounds that adversely affect both marine vertebrates and invertebrates and are reported to be responsible for human intoxication in aerosol form. In this work, a histopathological analysis accompanied by quantitative evaluation of tissue injury in mussels (Mytilus galloprovincialis) exposed to O. cf. ovata cells under natural and experimental conditions, provided baseline data on the health status of the mussels in terms of defensive and regressive pathological changes. We recorded a total of 15 health parameters in the digestive system, muscle, kidney and gills in mussels exposed to O. cf. ovata both in the laboratory and at sea. Animals exposed to different concentrations of O. cf. ovata cells (300, 500 and 1000cellsml(-)(1)) for 48h showed activation of the inflammatory response, which increased with the cell concentration, mainly characterized by haemocyte aggregates actively enclosing the algae, while mussel mortality was also recorded in some cases. Moreover the use of image analysis for the evaluation of digestive tubule damage revealed a pronounced increase in the lumen in terms of its area, perimeter and circularity, with a shift in a high percentage of tubules from an adsorbing profile to an atrophic profile. Animals collected from the natural environment during a summer bloom of O. cf. ovata in the Gulf of Naples (Italy) showed comparable lesions in terms of types and severity. This is the first quantitative study assessing damage to the digestive epithelia in terms of lumen modifications in mussels exposed to O. cf. ovata. The presented methodology provides a new technique for automating the evaluation of epithelial tubule modifications. Our results highlight the importance of monitoring the presence of O. cf. ovata in this area, taking into account the effects on the residing marine species.
Subject(s)
Mytilus/parasitology , Shellfish/parasitology , Animals , Dinoflagellida , Histological Techniques , Image Processing, Computer-Assisted , ItalyABSTRACT
In recent years, mass mortality events of benthic invertebrates in the Mediterranean Sea have been documented to coincide with the increased seawater temperatures associated with global climate change. Following a disease outbreak in gorgonians during the summer seasons of 2008 and 2009 in the Gulf of Naples (Tyrrhenian Sea), we conducted gross and microscopic analyses of healthy and diseased specimens of Eunicella cavolinii and E. singularis using both light and electron microscopy (SEM). Macroscopically, diseased colonies exhibited evident tissue thinning, and dead colonies showed a complete loss of polyps and coenenchyme, exposing their skeletons to settlement by fouling organisms. Histopathology revealed chronic inflammatory lesions at the polyp and axial level, characterized by amoebocyte infiltration of tissue accompanied by new apposition of melanin/gorgonin sheets. We interpreted this response as a defense against different kinds of pathogens-identified as mainly a heterogeneous consortium of filamentous cyanobacteria-and which gradually led to enlargement and hardening of the coral axis, which resembled a wood-like structure at the final stage of the disease. These processes elicited the formation of multiple inflammatory nodules and capsules, some of which were macroscopically visible. A parallel 16S rRNA and ITS analysis of the diseased tissue identified Synechococcus, Arthrospira and other uncultured cyanobacteria grouped within the Oscillatoriales. These results suggest that a cyanobacterial consortium is involved in the pathogenesis of the inflammatory disease leading to the mortality of Gorgoniaceae in the area. Finally, there were anomalously high temperatures (up to 25°C) between 10 and 20 m depth during the sampling period, particularly in June 2009. This supports the hypothesis that the coral skeleton may serve as a reservoir for the pathogens in cooler seasons, with warmer conditions leading to pathogen reactivation and recurring mortality events.
