Most cancers arise from oncogenic changes in the genomes of somatic cells, and while the cells may migrate by metastasis, they remain within that single individual. Natural transmission of cancer cells from one individual to another has been observed in two distinctive cases in mammals (Tasmanian devils1 and dogs2,3), but these are generally considered to be rare exceptions in nature. The discovery of transmissible cancer in soft-shell clams (Mya arenaria)4 suggested that this phenomenon might be more widespread. Here we analyzed disseminated neoplasia in mussels (Mytilus trossulus), cockles (Cerastoderma edule), and golden carpet shell clams (Polititapes aureus) and found that neoplasias in all three species are attributable to independent transmissible cancer lineages. In mussels and cockles, the cancer lineages are derived from their respective host species, but unexpectedly, cancer cells in P. aureus are all derived from Venerupis corrugata, a different species living in the same geographic area. No cases of disseminated neoplasia have thus far been found in V. corrugata from the same region. These findings show that transmission of cancer cells in the marine environment is common in multiple species, that it has originated many times, and that while most transmissible cancers were found spreading within the species of origin, cross-species transmission of cancer cells can occur.

-The genus Perkinsus includes protistan parasites infecting marine molluscs throughout the world, some of which are associated with mass mortalities. Life cycle involves vegetative proliferation within the host, by which a cell named trophozoite undergoes successive bipartitioning. Other stages have been observed in vitro or in vivo, depending on the species: hypnospore, zoosporangium and zoospore. Molecular taxonomy supports a close affinity between dinoflagellates and Perkinsus spp. Six species of Perkinsus are currently considered valid: P. marinus, P. olseni, P. qugwadi, P. chesapeaki, P. andrewsi and P. mediterraneus. Histology and, above all, incubation of host tissues in Ray's fluid thioglycollate medium (RFTM) are classic diagnostic methods. In addition, more sensitive and quicker molecular diagnostic techniques based on either immunoassays or PCR have been developed for Perkinsus spp. Epizootiological studies have shown a marked influence of water temperature and salinity on P. marinus infection in oysters Crassostrea virginica, thus determining parasite geographical range and temporal disease dynamics (seasonality). In vitro cultures have been established for four Perkinsus spp. Immune response to infection varies depending on host and involves phagocytosis or encapsulation of the parasite cells by host haemocytes. A polypeptide is secreted by clamTapes philippinarum haemocytes that could kill the parasite. In vitro cultured P. marinus cells secrete proteases that are likely involved in degradation of host tissues. P. marinus can suppress the toxic oxygen radicals produced by host haemocytes. In addition to host death, sublethal effects caused by Perkinsus spp. (reduction of fecundity, growth, and condition) may have significant ecological and economic implications. Various strategies have been assayed to mitigate the consequences of P. marinus epizootics on the oyster industry: modifications of management/culture procedures, selective breeding to obtain resistant oyster strains, and the use of triploid oysters and allochthonous oyster species. Some chemotherapeutants have been proved to inhibit or kill parasite cells in vitro.

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Morphological characters of zoosporulation stages and DNA sequence of the internal transcribed spacer (ITS) region and the small subunit ribosomal RNA (SSU rRNA) gene confirmed that the aetiological agent of perkinsosis in the clam Tapes decussatus from Galicia (NW Spain) was Perkinsus atlanticus Azevedo, 1989. In vitro modulation by temperature and salinity of the zoosporulation of the parasite was studied. The optimum temperature range for zoosporulation was 19 to 28°C. The temperature range allowing zoosporulation in vitro was 15 to 32°C, which is broader than previously reported (24 to 28°C) for P. atlanticus, and strongly suggests that zoospores can be produced in Galician Rías, where temperature ranges from 10 to 22°C. Prezoosporangia held at 10°C for 2 mo (similar to winter conditions in Galician waters) gave rise to viable zoospores after they were transferred to higher temperatures. This suggests that prezoosporangia could overwinter and zoosporulate in the next spring. Zoospores could survive for up to 22 and 14 d at 28 and 10°C, respectively. The optimum salinity range for zooporulation was 25 to 35 ‰. Zoospore production was abruptly reduced as salinity decreased. The lowest salinity at which zoosporulation was observed was 10 ‰. The effectiveness of different chlorine concentrations and exposure lengths to kill prezoosporangia and zoospores was tested. No survival of free zoospores, free prezoosporangia and prezoosporangia included in gill tissue was observed after incubation for 1 h with 50, 200 and 3000 ppm of chlorine, respectively.

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Transmissible cancers are parasitic malignant cell lineages that acquired the ability to infect new hosts from the same species, or sometimes related species. First described in dogs and Tasmanian devils, transmissible cancers were later discovered in some marine bivalves affected by a leukemia-like disease. In Mytilus mussels, two lineages of Bivalve Transmissible Neoplasia (BTN) have been described to date (MtrBTN1 and MtrBTN2), both emerged in a M. trossulus founder individual. Here, we performed an extensive screening of genetic chimerism, a hallmark of transmissible cancer, by genotyping 106 SNPs of 5907 European Mytilus mussels. The genetic analysis allowed us to simultaneously obtain the genotype of hosts -M. edulis, M. galloprovincialis or hybrids -and the genotype of tumors of heavily infected individuals. In addition, a subset of 222 individuals were systematically genotyped and analysed by histology in order to screen for possible non-transmissible cancers. We detected MtrBTN2 at low prevalence in M.edulis, and also in M. galloprovincialis and hybrids although at a much lower prevalence. No MtrBTN1 or new BTN were found, but 8 individuals with non-transmissible neoplasia were observed at a single polluted site on the same sampling date. We observed a diversity of MtrBTN2 genotypes that appeared more introgressed or more ancestral than MtrBTN1 and reference healthy M. trossulus individuals. The observed polymorphism is most likely due to somatic null alleles caused by structural variations or point mutations in primer-binding sites leading to enhanced detection of the host alleles. Despite low prevalence, two sublineages divergent by 10% fixed somatic null alleles and one non-synonymous mtCOI substitution, are co-spreading in the same geographic area, suggesting a complex diversification of MtrBTN2 since its emergence and host species shift.

Mediterranean mussels Mytilus galloprovincialis were experi~nentally cultured from 5 rafts located in 4 Galician Rias, following the established industrial procedure. Cultures were sampled monthly until mussels exceeded market slze. Observation of histological sections of sampled mussels by light microscopy demonstrated symbionts which could be classified into 3 groups according to their pathogenicity. The first group consisted of symbionts with unnoticeable pathogenic effects including: prokaryotic inclusion bodies (PlB) in digestive gland and gills, an unidentified protistan in digestive primary ducts, a kidney coccidian, intracytoplasmic ciliates in digestive tubules, gill ciliates and a turbellarian in the ~ntestinal lumen. The second group comprised syn~bionts that could damage the host, although unlikely to be lethal, including: the microsporidian Steinhausia mytilovum, the flatworm Urastoma cyprinae, and the copepod Mytilicola intestinahs. The third group included the protistan Marteilia refringens and the trematode Proctoeces rnaculatus, potentially lethal pathogens. In addition, mussels with haemocytic infiltration of tissues and granulocytomas and a few cases of disseminated neoplasia were detected. The qualitative composition of mussel symbiont community was similar at the 5 study sites, except for 3 symbionts which were not detected at some sites Quantitatively, symbiont loads were higher and histological signs of stress more abundant in Moana and Vilagarcia (the most inner sites in the Rias), intermediate in Illa de Arousa and Muros, and lower in Lorbe. Symbionts increased in prevalence as mussels grew. Some of the symbionts were detected in mussel seed at the beginning of the experimental cultures.