Carolyn S. Friedman

Climate Change Influences on Marine Infectious Diseases: Implications for Management and Society

Infectious diseases are common in marine environments, but the effects of a changing climate on marine pathogens are not well understood. Here we review current knowledge about how the climate drives host-pathogen interactions and infectious disease outbreaks. Climate-related impacts on marine diseases are being documented in corals, shellfish, finfish, and humans; these impacts are less clearly linked for other organisms. Oceans and people are inextricably linked, and marine diseases can both directly and indirectly affect human health, livelihoods, and well-being. We recommend an adaptive management approach to better increase the resilience of ocean systems vulnerable to marine diseases in a changing climate. Land-based management methods of quarantining, culling, and vaccinating are not successful in the ocean; therefore, forecasting conditions that lead to outbreaks and designing tools/approaches to influence these conditions may be the best way to manage marine disease.

Densovirus associated with sea-star wasting disease and mass mortality

Significance Sea stars inhabiting the Northeast Pacific Coast have recently experienced an extensive outbreak of wasting disease, leading to their degradation and disappearance from many coastal areas. In this paper, we present evidence that the cause of the disease is transmissible from disease-affected animals to apparently healthy individuals, that the disease-causing agent is a virus-sized microorganism, and that the best candidate viral taxon, the sea star-associated densovirus (SSaDV), is in greater abundance in diseased than in healthy sea stars. Populations of at least 20 asteroid species on the Northeast Pacific Coast have recently experienced an extensive outbreak of sea-star (asteroid) wasting disease (SSWD). The disease leads to behavioral changes, lesions, loss of turgor, limb autotomy, and death characterized by rapid degradation (“melting”). Here, we present evidence from experimental challenge studies and field observations that link the mass mortalities to a densovirus (Parvoviridae). Virus-sized material (i.e., <0.2 μm) from symptomatic tissues that was inoculated into asymptomatic asteroids consistently resulted in SSWD signs whereas animals receiving heat-killed (i.e., control) virus-sized inoculum remained asymptomatic. Viral metagenomic investigations revealed the sea star-associated densovirus (SSaDV) as the most likely candidate virus associated with tissues from symptomatic asteroids. Quantification of SSaDV during transmission trials indicated that progression of SSWD paralleled increased SSaDV load. In field surveys, SSaDV loads were more abundant in symptomatic than in asymptomatic asteroids. SSaDV could be detected in plankton, sediments and in nonasteroid echinoderms, providing a possible mechanism for viral spread. SSaDV was detected in museum specimens of asteroids from 1942, suggesting that it has been present on the North American Pacific Coast for at least 72 y. SSaDV is therefore the most promising candidate disease agent responsible for asteroid mass mortality.

OCEAN WARMING EFFECTS ON GROWTH, REPRODUCTION, AND SURVIVORSHIP OF SOUTHERN CALIFORNIA ABALONE

Traditional fisheries management in southern California has failed, in part because it is based on an assumption of an unvarying environment and is focused on size limits rather than insuring the persistence of aggregations of large fecund individuals. The combined effect of low frequency climatic variability and anthropogenic perturbations can have dramatic consequences for abalone in southern California. Abalone species are tightly linked to kelp forest ecosystems that, besides furnishing habitat, also provide the main food source for abalone. In southern California, kelp canopies are very sensitive to oceanographic climate because the kelp depend upon high nutrients in the water column. Oceanic warming, in turn, results in decreased nutrients in the surface water, and this is correlated with marked reductions in giant kelp biomass. Here we address the additive effects of ocean warming on two species of California abalone (the red abalone, Haliotis rufescens; and the green abalone, H. fulgens) by subjecting them to varied environmental conditions similar to cool, normal, and warm phases of the California current in the southern California Bight. Our experimental design simultaneously tested the synergistic effects of temperature and food quantity and quality on survivorship, growth, and reproduction. For red abalone, warm temperatures increased the onset of with- ering syndrome, a fatal abalone disease, and halted growth and reproduction. In contrast, green abalone survivorship, growth, and reproduction were relatively robust irrespective of temperature, while their growth and reproduction were most strongly influenced by food quantity. We found clear evidence suggesting that, combined with overfishing, California abalone populations are adversely affected by ecosystem responses to ocean warming: Cool- water red abalone suffer stronger consequences in warm water than do green abalone. Conservation, restoration, and recovery plans of remnant California abalone populations

Characterizing the metabolic actions of natural stresses in the California red abalone, Haliotis rufescens using 1H NMR metabolomics

Withering syndrome in California red abalone (Haliotis rufescens) is caused by the Rickettsiales-like prokaryote (WS-RLP) Candidatus Xenohaliotis californiensis. WS-RLP infection is not sufficient to cause withering syndrome, and for reasons not yet well understood additional stressors such as elevated water temperature appear to influence disease development. Using nuclear magnetic resonance (NMR) based metabolomics, we have investigated the influence of food availability, temperature, and bacterial infection, both individually and in combination, on the metabolic status of the red abalone. Food limitation caused dramatic reductions in all observed classes of foot muscle metabolites, while at the same time metabolite levels within the digestive gland were preserved or increased. We also found that food limitation in combination with elevated temperature led to greater metabolic perturbations in both tissue types than those observed under food limitation alone. WS-RLP infection and food-limitation resulted in many of the same metabolic changes within the tissues studied, although the effects of infection were less severe. We observed increased levels of homarine in the digestive gland of both food-limited and WS-RLP-infected animals, yet only observed increased homarine levels in the foot muscle of infected abalone. These results further support the recently established glucose-to-homarine ratio in foot muscle as a potential marker for differentiating WS-RLP-infected animals from those of both healthy and food limited abalone. Furthermore, we found that the NMR metabolic data correlates well with histological measurements supporting the use of the metabolomics approach for characterizing both normal and pathological events in marine species, particularly during periods of environmentally relevant stress.

SUMMER SEED MORTALITY OF THE PACIFIC OYSTER, CRASSOSTREA GIGAS THUNBERG GROWN IN TOMALES BAY, CALIFORNIA, USA: THE INFLUENCE OF OYSTER STOCK, PLANTING TIME, PATHOGENS, AND ENVIRONMENTAL STRESSORS

Abstract Summer seed mortality (SSM) has occurred yearly in Tomales Bay, California since 1993. SSM has resulted in up to 90% cumulative losses, and has been associated with extreme temperature, phytoplankton blooms, and an oyster herpesvirus. In this study, three stocks of Pacific oysters were planted at three sites in California (Inner Tomales Bay, Outer Tomales Bay, and Bodega Harbor) in October of 2000 (Fall) and April of 2001 (Spring) and monitored for mortality, growth, and health status. In April of 2001, a similar study was conducted in Totten Inlet, WA state using cohorts of oysters planted in California; animals were monitored for mortality and growth. Temperature data were collected at all sites; phytoplankton abundance data were collected at the California sites. Mortality occurred only at the Inner Tomales Bay site where losses were correlated with maximum temperatures (r = 0.949) and preferentially affected faster growing oysters (r = 0.916). Significant differences in cumulative mortality were identified among oysters stocks and two of the three oysters stocks planted in the fall outperformed their cohorts planted in the spring (P < 0.0001). Microscopic changes in connective tissue and digestive tubules are consistent with previous observations of herpesvirus infections in oysters including: diffuse to multifocal pertibular hemocyte infiltration, diapedesis, dilation of the digestive tubules, nuclear hypertrophy, and chromatin margination. Nuclear hypertrophy and chromatin margination, in particular, are suggestive of herpesvirus infections; these histological changes were rare indicating the need to use multiple diagnostic methods when oyster herpesviruses are suspected to cause SSM. Temperature maxima (∼25°C) experienced at the Inner Tomales Bay site are not considered extreme for Pacific oyster survival; the association between oyster herpesviruses and temperature in Tomales Bay, California is discussed.

Nocardia crassostreae sp. nov., the causal agent of nocardiosis in Pacific oysters.

Seven strains of bacteria were isolated from Pacific oysters, Crassostrea gigas, with a focal or systemic disease. The strains were aerobic, Gram-positive, acid-fast, produced a mycelium which fragmented into irregular rod-like elements, had a peptidoglycan containing meso-diaminopimelic acid, arabinose and galactose as major sugars, mycolic acids with 46-58 carbon atoms and G + C-rich DNA. All of these properties are consistent with the classification of the organisms in the genus Nocardia. A partial sequence of the 16S rRNA gene of isolate NB4H was determined following isolation and cloning of the PCR-amplified gene. The sequence was aligned with those of representative mycolic-acid-containing taxa and a phylogenetic tree was generated using the neighbour-joining method. It was evident from the phylogenetic tree that the three strains tested, RB1, OB3P and NB4H, were identical and belonged to the Nocardia otitidiscaviarum rRNA sub-group. The biochemical, chemical, morphological and physiological properties of the isolates were also essentially identical and served to distinguish them from representative nocardiae. It is, therefore, proposed that the strains isolated from the diseased Pacific oysters be assigned to a new species, Nocardia crassostreae. The type strain is NB4H (= ATCC 700418).

Detection of the oyster herpesvirus in commercial bivalves in northern California, USA: conventional and quantitative PCR

The ostreid herpesvirus (OsHV-1) and related oyster herpesviruses (OsHV) are associated with world-wide mortalities of larval and juvenile bivalves. To quantify OsHV viral loads in mollusc tissues, we developed a SYBR Green quantitative PCR (qPCR) based on the A-region of the OsHV-1 genome. Reaction efficiency and precision were demonstrated using a plasmid standard curve. The analytical sensitivity is 1 copy per reaction. We collected Crassostrea gigas, C. sikamea, C. virginica, Ostrea edulis, O. lurida, Mytilus galloprovincialis, and Venerupis phillipinarum from Tomales Bay (TB), and C. gigas from Drakes Estero (DE), California, U.S.A., and initially used conventional PCR (cPCR) to test for presence of OsHV DNA. Subsequently, viral loads were quantified in selected samples of all tested bivalves except O. lurida. Copy numbers were low in each species tested but were significantly greater in C. gigas (p < 0.0001) compared to all other species, suggesting a higher level of infection. OsHV DNA was detected with cPCR and/or qPCR and confirmed by sequencing in C. gigas, C. sikamea, C. virginica, O. edulis, M. galloprovincialis, and V phillipinarum from TB and C. gigas from DE. These data indicate that multiple bivalve species may act as reservoirs for OsHV in TB. A lack of histological abnormalities in potential reservoirs requires alternative methods for their identification. Further investigation is needed to determine the host-parasite relationship for each potential reservoir, including characterization of viral loads and their relationship with infection (via in situ hybridization), assessments of mortality, and host responses.

Gene expression patterns of abalone, Haliotis tuberculata, during successive infections by the pathogen Vibrio harveyi.

Since 1998, episodic mass mortality of the abalone Haliotistuberculata has been observed along the northern Brittany coast of France caused by a complex interaction among the host, pathogen and environmental factors. In the present study, abalone were submitted to two successive infections with the pathogen Vibrioharveyi under controlled conditions. During the first challenge, infection by V.harveyi resulted in 64% mortality of mature abalone. After a second infection of those surviving the first challenge, only 44% mortality was observed. Physiological variability in the host response appears to be a major determinant in susceptibility to V.harveyi. In order to isolate differentially expressed genes in H.tuberculata challenged with this bacterium, suppression subtractive hybridization (SSH) cDNA libraries were constructed from muscle of moribund abalone (susceptibles), surviving individuals (apparently resistant to the bacterium) and control (unexposed) animals. Of the 1152 clones sequenced, 218 different partial cDNA sequences were obtained and represented 69 known genes. Of these, 65 were identified for the first time in H.tuberculata. Using real-time PCR, a time-course study was conducted on 19 of the genes identified by SSH. A majority of differentially expressed transcripts were down-regulated in susceptible individuals as compared to their resistant counterparts. Bacterial challenge of abalone resulted in the up-regulation of three transcripts (encoding ferritin, heat shock protein HSP84 and fatty acid binding protein FABP) in those that survived exposure to V.harveyi. This study has identified potential candidates for further investigation into the functional basis of resistance and susceptibility to summer vibriosis outbreaks in abalone.

Managing marine disease emergencies in an era of rapid change

Infectious marine diseases can decimate populations and are increasing among some taxa due to global change and our increasing reliance on marine environments. Marine diseases become emergencies when significant ecological, economic or social impacts occur. We can prepare for and manage these emergencies through improved surveillance, and the development and iterative refinement of approaches to mitigate disease and its impacts. Improving surveillance requires fast, accurate diagnoses, forecasting disease risk and real-time monitoring of disease-promoting environmental conditions. Diversifying impact mitigation involves increasing host resilience to disease, reducing pathogen abundance and managing environmental factors that facilitate disease. Disease surveillance and mitigation can be adaptive if informed by research advances and catalysed by communication among observers, researchers and decision-makers using information-sharing platforms. Recent increases in the awareness of the threats posed by marine diseases may lead to policy frameworks that facilitate the responses and management that marine disease emergencies require.

Bacterial diseases in marine bivalves.

Bivalve aquaculture is seriously affected by many bacterial pathogens that cause high losses in hatcheries as well as in natural beds. A number of Vibrio species, but also members of the genera Nocardia and Roseovarius, are considered important pathogens in aquaculture. The present work provides an updated overview of main diseases and implicated bacterial species affecting bivalves. This review focuses on aetiological agents, their diversity and virulence factors, the diagnostic methods available as well as information on the dynamics of the host-parasite relationship.