Date Published: December 18, 2013
Publisher: Public Library of Science
Author(s): Fernanda F. M. Mazzillo, Karen Shapiro, Mary W. Silver, Ira J. Blader.
Toxoplasma gondii is a land-derived parasite that infects humans and marine mammals. Infections are a significant cause of mortality for endangered southern sea otters (Enhydra lutris nereis), but the transmission mechanism is poorly understood. Otter exposure to T. gondii has been linked to the consumption of marine turban snails in kelp (Macrocystis pyrifera) forests. It is unknown how turban snails acquire oocysts, as snails scrape food particles attached to surfaces, whereas T. gondii oocysts enter kelp beds as suspended particles via runoff. We hypothesized that waterborne T. gondii oocysts attach to kelp surfaces when encountering exopolymer substances (EPS) forming the sticky matrix of biofilms on kelp, and thus become available to snails. Results of a dietary composition analysis of field-collected snails and of kelp biofilm indicate that snails graze the dense kelp-biofilm assemblage composed of pennate diatoms and bacteria inserted within the EPS gel-like matrix. To test whether oocysts attach to kelp blades via EPS, we designed a laboratory experiment simulating the kelp forest canopy in tanks spiked with T. gondii surrogate microspheres and controlled for EPS and transparent exopolymer particles (TEP – the particulate form of EPS). On average, 19% and 31% of surrogates were detected attached to kelp surfaces covered with EPS in unfiltered and filtered seawater treatments, respectively. The presence of TEP in the seawater did not increase surrogate attachment. These findings support a novel transport mechanism of T. gondii oocysts: as oocysts enter the kelp forest canopy, a portion adheres to the sticky kelp biofilms. Snails grazing this biofilm encounter oocysts as ‘bycatch’ and thereby deliver the parasite to sea otters that prey upon snails. This novel mechanism can have health implications beyond T. gondii and otters, as a similar route of pathogen transmission may be implicated with other waterborne pathogens to marine wildlife and humans consuming biofilm-feeding invertebrates.
Toxoplasma gondii is a coccidian parasite that infects humans and warm-blooded animals . Infections with this terrestrial parasite have also been documented in marine mammals , including the southern sea otter (Enhydra lutris neries). The southern sea otter is an endangered subspecies that inhabits giant kelp forests, including those of Macrocystis pyrifera, along the California coast. Toxoplasma gondii is recognized as a significant cause of mortality in southern sea otters, with harmful consequences for the health and recovery of this population . Infected otters have been detected at several locations along the coast of California, and high-risk sites for otter exposure were described in populations from Morro Bay and Cambria, California , .
Two experiments were designed to: (1) provide qualitative data on snail diet and kelp biofilm composition; and (2) test whether T. gondii oocyst surrogates adhere to kelp blade surfaces via EPS on the blades and/or via TEP suspended in the water that subsequently become biofilm on the blades. Together these experiments examine the possible mechanism(s) by which T. gondii oocysts may become associated with kelp surfaces and whether snails can consume organisms entrained within the kelp biofilm: if such associations are found, then a delivery route of the planktonic oocysts to the endangered otter would be identified for the first time. We considered both EPS (the matrix of the biofilm on kelp surfaces) and TEP in the water, as potential agents that could deliver Toxoplasma to snails that graze the kelp surfaces with biofilms. Dragon green microspheres that have been previously validated as surrogate particles for T. gondii oocysts were used because they have surface properties (i.e., size, specific gravity, hydrophobicity, and surface charge) that resemble those of T. gondii oocysts . Due to the biohazardous nature of T. gondii oocysts, employing surrogates in mesocosm experiments provides an alternative approach for evaluating the waterborne transport of this zoonotic pathogen. Previous studies have successfully applied these surrogates to demonstrate waterborne transport of oocysts  and their association with macroaggregates .
The goal of the present study was to test whether extracellular polymer substances (EPS), which form the matrix of biofilms colonizing the surfaces of M. pyrifera blades, play a role in the transmission of the protozoan parasite T. gondii to benthic feeding turban snails. Marine turban snails have been implicated in the exposure of southern sea otters to T. gondii. As this parasite has been identified as a significant cause of mortality in endangered southern sea otter populations, it is critically important to understand the transmission mechanisms to otters, so that prevention or management strategies can be developed to reduce likelihood of exposure. Here we show that T. gondii surrogates may adhere to the biofilm that colonizes the surfaces of kelp blades, thereby becoming available to turban snails that feed upon organisms associated with this biofilm.
Our findings suggest a novel route of exposure of sea otters to the protozoan parasite T. gondii. Although the estimated number of T. gondii oocysts that are transported to kelp forests is unknown, these experimental results provide a mechanism to explain the transmission of T. gondii oocysts to sea otters. As T. gondii oocysts are deposited in the coastal ocean via contaminated runoff, we propose that a proportion of them attach to the sticky EPS biofilms on the kelp blades, with the surface communities composed in part of benthic diatoms and bacteria. Snails, which feed by scraping these benthic diatoms from the surface of kelp blade using their radula, would ingest T. gondii oocysts as ‘bycatch’, explaining why sea otters that specialize on consuming subtidal snails are more likely to be exposed to this parasite. The route of infection of other pathogens to marine wildlife may also occur via an EPS-adhesion mechanism such as that described here. Insight into EPS-mediated pathogen transmission may also have significant implications for human public health, due to consumption of marine animals that feed on EPS-coated substances . This study, therefore, suggests a new transmission route for delivering microscopic-sized pelagic pathogens to higher trophic level predators in marine ecosystems.