Porphyrin detection in denatured cnidarian tissue extracts

Porphyrin metabolic disruption from exposure to xenobiotic contaminants such as heavy metals, dioxins, and aromatic hydrocarbons can elicit overproduction of porphyrins. Measurement of porphyrin levels, when used in conjunction with other diagnostic assays, can help elucidate an organism’s physiological condition and provide evidence for exposure to certain toxicants. A sensitive microplate fluorometric assay has been optimized for detecting total porphyrin levels in detergent solubilized protein extracts from symbiotic, dinoflagellate containing cnidarian tissues. The denaturing buffer used in this modified assay contains a number of potentially interfering components (e.g., sodium dodecyl sulfate (SDS), dithiothreitol (DTT), protease inhibitors, and chlorophyll from the symbiotic zooxanthellae), which required examination and validation. Examination of buffer components were validated for use in this porphyrin assay; while the use of a specific spectrofluorometric filter (excitation 400 ± 15 nm; emission 600 ± 20 nm) minimized chlorophyll interference. The detection limit for this assay is 10 fmol of total porphyrin per μg of total soluble protein and linearity is maintained up to 5000 fmol. The ability to measure total porphyrins in a SDS protein extract now allows a single extract to be used in multiple assays. This is an advantage over classical methods, particularly when tissue samples are limiting, as is often the case with coral due to availability and collection permit restrictions.

Deep coral and associated species taxonomy and ecology: (DeepCAST) II Expedition Report, Roatan, Honduras, May 21-28, 2011

NOAA has a mandate to explore and understand deep-sea coral ecology under Magnuson-Stevens Sustainable Fisheries Conservation Act Reauthorization of 2009. Deep-sea corals are increasingly considered a proxy for marine biodiversity in the deep-sea because corals create complex structure, and this structure forms important habitat for associated species of shrimp, crabs, sea stars, brittle stars, and fishes. Yet, our understanding of the nature of the relationships between deep-corals and their associated species is incomplete. One of the primary challenges of conducting any type of deep-sea coral (DSC) research is access to the deep-sea. The deep-sea is a remote environment that often requires long surface transits and sophisticated research vehicles like submersibles and remotely operated vehicles (ROVs). The research vehicles often require substantial crew, and the vehicles are typically launched from large research vessels costing many thousands of dollars a day. To overcome the problem of access to the deep-sea, the Deep Coral and Associated Species Taxonomy and Ecology (DeepCAST) Expeditions are pioneering the use of shore-based submersibles equipped to do scientific research. Shore-based subs alleviate the need for expensive ships because they launch and return under their own power. One disadvantage to the approach is that shore-based subs are restricted to nearby sites. The disadvantage is outweighed, however, by the benefit of repeated observations, and the opportunity to reduce the costs of exploration while expanding knowledge of deep-sea coral ecology.

A survey of deep-water coral and sponge habitats along the west coast of the US using a remotely operated vehicle: NOAA Fisheries Survey Vessel (FSV)'Bell M. Shimada', November 1-5, 2010

Remotely operated vehicle (ROV) surveys were conducted from NOAA’s state-of-the-art Fisheries Survey Vessel (FSV) Bell M. Shimada during a six-day transit November 1-5, 2010 between San Diego, CA and Seattle, WA. The objective of this survey was to locate and characterize deep-sea coral and sponge ecosystems at several recommended sites in support of NOAA’s Coral Reef Conservation Program. Deep-sea corals and sponges were photographed and collected whenever possible using the Southwest Fisheries Science Center’s (SWFSC) Phantom ROV ‘Sebastes’ (Fig. 1). The surveyed sites were recommended by National Marine Sanctuary (NMS) scientists at Monterey Bay NMS, Gulf of the Farallones NMS, and Olympic Coast NMS (Fig. 2). The specific sites were: Sur Canyon, The Football, Coquille Bank, and Olympic Coast NMS. During each dive, the ROV collected digital still images, video, navigation, and along-track conductivity-temperature-depth (CTD), and optode data. Video and high-resolution photographs were used to quantify abundance of corals, sponges, and associated fishes and invertebrates to the lowest practicable taxonomic level, and also to classify the seabed by substrate type. A reference laser system was used to quantify area searched and estimate the density of benthic fauna.