Saline-saturated DMSO-EDTA as a storage medium for microbial DNA analysis from coral mucus swab samples

The mucus surface layer of corals plays a number of integral roles in their overall health and fitness. This mucopolysaccharide coating serves as vehicle to capture food, a protective barrier against physical invasions and trauma, and serves as a medium to host a community of microorganisms distinct from the surrounding seawater. In healthy corals the associated microbial communities are known to provide antibiotics that contribute to the coral’s innate immunity and function metabolic activities such as biogeochemical cycling. Culture-dependent (Ducklow and Mitchell, 1979; Ritchie, 2006) and culture-independent methods (Rohwer, et al., 2001; Rohwer et al., 2002; Sekar et al., 2006; Hansson et al., 2009; Kellogg et al., 2009) have shown that coral mucus-associated microbial communities can change with changes in the environment and health condition of the coral. These changes may suggest that changes in the microbial associates not only reflect health status but also may assist corals in acclimating to changing environmental conditions. With the increasing availability of molecular biology tools, culture-independent methods are being used more frequently for evaluating the health of the animal host. Although culture-independent methods are able to provide more in-depth insights into the constituents of the coral surface mucus layer’s microbial community, their reliability and reproducibility rely on the initial sample collection maintaining sample integrity. In general, a sample of mucus is collected from a coral colony, either by sterile syringe or swab method (Woodley, et al., 2008), and immediately placed in a cryovial. In the case of a syringe sample, the mucus is decanted into the cryovial and the sealed tube is immediately flash-frozen in a liquid nitrogen vapor shipper (a.k.a., dry shipper). Swabs with mucus are placed in a cryovial, and the end of the swab is broken off before sealing and placing the vial in the dry shipper. The samples are then sent to a laboratory for analysis. After the initial collection and preservation of the sample, the duration of the sample voyage to a recipient laboratory is often another critical part of the sampling process, as unanticipated delays may exceed the length of time a dry shipper can remain cold, or mishandling of the shipper can cause it to exhaust prematurely. In remote areas, service by international shipping companies may be non-existent, which requires the use of an alternative preservation medium. Other methods for preserving environmental samples for microbial DNA analysis include drying on various matrices (DNA cards, swabs), or placing samples in liquid preservatives (e.g., chloroform/phenol/isoamyl alcohol, TRIzol reagent, ethanol). These methodologies eliminate the need for cold storage, however, they add expense and permitting requirements for hazardous liquid components, and the retrieval of intact microbial DNA often can be inconsistent (Dawson, et al., 1998; Rissanen et al., 2010). A method to preserve coral mucus samples without cold storage or use of hazardous solvents, while maintaining microbial DNA integrity, would be an invaluable tool for coral biologists, especially those in remote areas. Saline-saturated dimethylsulfoxide-ethylenediaminetetraacetic acid (20% DMSO-0.25M EDTA, pH 8.0), or SSDE, is a solution that has been reported to be a means of storing tissue of marine invertebrates at ambient temperatures without significant loss of nucleic acid integrity (Dawson et al., 1998, Concepcion et al., 2007). While this methodology would be a facile and inexpensive way to transport coral tissue samples, it is unclear whether the coral microbiota DNA would be adversely affected by this storage medium either by degradation of the DNA, or a bias in the DNA recovered during the extraction process created by variations in extraction efficiencies among the various community members. Tests to determine the efficacy of SSDE as an ambient temperature storage medium for coral mucus samples are presented here.

Ecological Condition of Coastal Ocean Waters within Stellwagen Bank National Marine Sanctuary: 2008

In June 2008, the NOAA National Ocean Service (NOS), in conjunction with the EPA National Health and Environmental Effects Laboratory (NHEERL), conducted an assessment of the status of ecological condition of soft-bottom habitat and overlying waters within the boundaries of Stellwagen Bank National Marine Sanctuary (SBNMS). The sanctuary lies approximately 20 nautical miles east of Boston, MA in the southwest Gulf of Maine between Cape Ann and Cape Cod and encompassing 638 square nautical miles (2,181 km2). A total of 30 stations were targeted for sampling using standard methods and indicators applied in prior NOAA coastal studies and EPA’s Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA). A key feature adopted from these studies was the incorporation of a random probabilistic sampling design. Such a design provides a basis for making unbiased statistical estimates of the spatial extent of ecological condition relative to various measured indicators and corresponding thresholds of concern. Indicators included multiple measures of water quality, sediment quality, and biological condition (benthic fauna, fish tissue contaminant levels). Depths ranged from 31 – 137 m throughout the study area. About 76 % of the area had sediments composed of sands (< 20 % silt-clay), 17 % of the area was composed of intermediate muddy sands (20 – 80 % silt-clay), and 7 % of the sampled area consisted of mud (> 80 % siltclay). About 70 % of the area (represented by 21 sites) had sediment total organic carbon (TOC) concentrations < 5 mg/g and all but one site (located in Stellwagen Basin) had levels of TOC < 20 mg/g, which is well below the range potentially harmful to benthic fauna (> 50 mg/g). Surface salinities ranged from 30.6 – 31.5 psu, with the majority of the study region (approximately 80 % of the area) having surface salinities between 30.8 and 31.4 psu. Bottom salinities varied between 32.1 and 32.5 psu, with bottom salinities at all sites having values above the range of surface salinities. Surface-water temperatures varied between 12.1 and 16.8 ºC, while near-bottom waters ranged in temperature from 4.4 – 6.2 ºC. An index of density stratification (Δσt) indicated that the waters of SBNMS were stratified at the time of sampling. Values of Δσt at 29 of the 30 sites sampled in this study (96.7 % of the study area) varied from 2.1 – 3.2, which is within the range considered to be indicative of strong vertical stratification (Δσt > 2) and typical of the western Gulf of Maine in summer. Levels of dissolved oxygen (DO) were confined to a fairly narrow range in surface (8.8 – 10.4 mg/L) and bottom (8.5 – 9.6 mg/L) waters throughout the survey area. These levels are within the range considered indicative of good water quality (> 5 mg/L) with respect to DO. None of these waters had DO at low levels (< 2 mg/L) potentially harmful to benthic fauna and fish.

A baseline assessment of the ecological resources of Jobos Bay, Puerto Rico

This baseline assessment of Jobos Bay and surrounding marine ecosystems consists of a two part series. The first report (Zitello et al., 2008) described the characteristics of the Bay and its watershed, including modeling work related to nutrients and sediment fluxes, based on existing data. The second portion of this assessment, presented in this document, presents the results of new field studies conducted to fill data gaps identified in previous studies, to provide a more complete characterization of Jobos Bay and the surrounding coral reef ecosystems. Specifically, the objective was to establish baseline values for the distribution of habitats, nutrients, contaminants, fi sh, and benthic communities. This baseline assessment is the first step in evaluating the effectiveness in changes in best management practices in the watershed. This baseline assessment is part of the Conservation Effects Assessment Project (CEAP), which is a multi-agency effort to quantify the environmental benefits of conservation practices used by agricultural producers participating in selected U.S. Department of Agriculture (USDA) conservation programs. Partners in the CEAP Jobos Bay Special Emphasis Watershed (SEW) included USDA’s Agricultural Research Service (ARS) and the Natural Resources Conservation Service (NRCS), National Oceanic and Atmospheric Administration (NOAA) and the Government of Puerto Rico. The project originated from an on-going collaboration between USDA and NOAA on the U.S. Coral Reef Task Force. The Jobos Bay watershed was chosen because the predominant land use is agriculture, including agricultural lands adjacent to the Jobos Bay National Estuarine Research Reserve (JBNERR or Reserve), one of NOAA’s 26 National Estuarine Research Reserves (NERR). This report is organized into six chapters that represent a suite of interrelated studies. Chapter 1 provides a short introduction to Jobos Bay, including the land use and hydrology of the watershed. Chapter 2 is focused on benthic mapping and provides the methods and results of newly created benthic maps for Jobos Bay and the surrounding coral reef ecosystem. Chapter 3 presents the results of new surveys of fish, marine debris, and reef communities of the system. Chapter 4 is focused on the distribution of chemical contaminants in sediments within the Bay and corals outside of the Bay. Chapter 5 focuses on quantifying nutrient and pesticide concentrations in the surface waters at the Reserve’s System-Wide Monitoring Program (SWMP) sites. Chapter 6 is a brief summary discussion that highlights key findings of the entire suite of studies.

A biogeographic assessment of the Samoan Archipelago

This report examines the marine biogeography of the Samoan Archipelago (~14º S latitude along the international date-line) with a focus on regional ocean climate, connectivity among islands due to larval transport, distributions of reef fish and coral communities, and the extent of existing marine protected areas. Management decisions and prior assessments in the archipelago have typically been split along the international political boundary between the islands of Samoa and those of American Samoa despite their close proximity and shared resources. A key goal in this assessment was to compile data from both jurisdictions and to conduct the characterization across the entire archipelago. The report builds upon earlier assessments by re-analyzing and interpreting many pre-existing datasets, adding more recent biogeographic data sources, and by combining earlier findings into a multidisciplinary summary of marine biogeography. The assessment is divided into 5 chapters and supporting appendices. Each chapter was written and reviewed in collaboration with subject matter specialists and local experts. In Chapter 1, a short introduction to the overall scope and approach of the report is provided. In Chapter 2, regional ocean climate is characterized using remote sensing datasets and discussed in the context of local observations. In Chapter 3, regional ocean currents and transport of coral and fish larvae are investigated among the islands of the archipelago and surrounding island nations. In Chapter 4, distinct reef fish and coral communities across the archipelago are quantified on the basis of overall biodiversity, abundance, and community structure. In Chapter 5, the existing network of MPAs in American Samoa is evaluated based on the habitats, reef fish, and coral communities that are encompassed. Appendices provide analytical details omitted from some chapters for brevity as well as supplemental datasets needed as inputs for the main chapters in the assessment. Appendices include an inventory of regional seamounts, a description of shore to shelf edge benthic maps produced for Tutuila, analytical details of reef fish and coral datasets, and supplemental information on the many marine protected areas in American Samoa.

Assessment of existing information on Atlantic coastal fish habitats: development of a web-based spatial bibliography, query tools, and data summaries

The primary objective of this project, “the Assessment of Existing Information on Atlantic Coastal Fish Habitat”, is to inform conservation planning for the Atlantic Coastal Fish Habitat Partnership (ACFHP). ACFHP is recognized as a Partnership by the National Fish Habitat Action Plan (NFHAP), whose overall mission is to protect, restore, and enhance the nation’s fish and aquatic communities through partnerships that foster fish habitat conservation. This project is a cooperative effort of NOAA/NOS Center for Coastal Monitoring and Assessment (CCMA) Biogeography Branch and ACFHP. The Assessment includes three components; 1. a representative bibliographic and assessment database, 2. a Geographical Information System (GIS) spatial framework, and 3. a summary document with description of methods, analyses of habitat assessment information, and recommendations for further work. The spatial bibliography was created by linking the bibliographic table developed in Microsoft Excel and exported to SQL Server, with the spatial framework developed in ArcGIS and exported to GoogleMaps. The bibliography is a comprehensive, searchable database of over 500 selected documents and data sources on Atlantic coastal fish species and habitats. Key information captured for each entry includes basic bibliographic data, spatial footprint (e.g. waterbody or watershed), species and habitats covered, and electronic availability. Information on habitat condition indicators, threats, and conservation recommendations are extracted from each entry and recorded in a separate linked table. The spatial framework is a functional digital map based on polygon layers of watersheds, estuarine and marine waterbodies derived from NOAA’s Coastal Assessment Framework, MMS/NOAA’s Multipurpose Marine Cadastre, and other sources, providing spatial reference for all of the documents cited in the bibliography. Together, the bibliography and assessment tables and their spatial framework provide a powerful tool to query and assess available information through a publicly available web interface. They were designed to support the development of priorities for ACFHP’s conservation efforts within a geographic area extending from Maine to Florida, and from coastal watersheds seaward to the edge of the continental shelf. The Atlantic Coastal Fish Habitat Partnership has made initial use of the Assessment of Existing Information. Though it has not yet applied the AEI in a systematic or structured manner, it expects to find further uses as the draft conservation strategic plan is refined, and as regional action plans are developed. It also provides a means to move beyond an “assessment of existing information” towards an “assessment of fish habitat”, and is being applied towards the National Fish Habitat Action Plan (NFHAP) 2010 Assessment. Beyond the scope of the current project, there may be application to broader initiatives such as Integrated Ecosystem Assessments (IEAs), Ecosystem Based Management (EBM), and Marine Spatial Planning (MSP).

Ecological condition of coastal ocean waters along the U.S. Mid-Atlantic Bight: 2006

In May 2006, the NOAA National Ocean Service (NOS), in conjunction with the EPA National Health and Environmental Effects Laboratory (NHEERL), conducted an assessment of the status of ecological condition of soft-bottom habitat and overlying waters throughout the mid-Atlantic Bight (MAB) portion of the eastern U.S. continental shelf. The study area encompassed the region from Cape Cod, MA and Nantucket Shoals in the northeast to Cape Hatteras in the south, and was defined using a one nautical mile buffer of the shoreline extended seaward to the shelf break (~100-m depth contour). A total of 50 stations were targeted for sampling using standard methods and indicators applied in prior NOAA coastal studies and EPA’s Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA). A key feature adopted from these studies was the incorporation of a random probabilistic sampling design. Such a design provides a basis for making unbiased statistical estimates of the spatial extent of ecological condition relative to various measured indicators and corresponding thresholds of concern. Indicators included multiple measures of water quality, sediment quality, and biological condition (benthic fauna). Through coordination with the NOAA Fisheries Service/Northeast Fisheries Science Center (NFS/NEFSC), samples of summer flounder (Paralichthys dentatus) also were obtained from 30 winter 2007 bottom-trawl survey stations in overlapping portions of the study area and used for analysis of chemical-contaminant body burdens.

Shallow-water benthic habitats of St. John, U.S. Virgin Islands

Coral reef ecosystems of the Virgin Islands Coral Reef National Monument, Virgin Islands National Park and the surrounding waters of St. John, U.S. Virgin Islands are a precious natural resource worthy of special protection and conservation. The mosaic of habitats including coral reefs, seagrasses and mangroves, are home to a diversity of marine organisms. These benthic habitats and their associated inhabitants provide many important ecosystem services to the community of St. John, such as fishing, tourism and shoreline protection. However, coral reef ecosystems throughout the U.S. Caribbean are under increasing pressure from environmental and anthropogenic stressors that threaten to destroy the natural heritage of these marine habitats. Mapping of benthic habitats is an integral component of any effective ecosystem-based management approach. Through the implementation of a multi-year interagency agreement, NOAA’s Center for Coastal Monitoring and Assessment - Biogeography Branch and the U.S. National Park Service (NPS) have completed benthic habitat mapping, field validation and accuracy assessment of maps for the nearshore marine environment of St. John. This work is an expansion of ongoing mapping and monitoring efforts conducted by NOAA and NPS in the U.S. Caribbean and replaces previous NOAA maps generated by Kendall et al. (2001) for the waters around St. John. The use of standardized protocols enables the condition of the coral reef ecosystems around St. John to be evaluated in context to the rest of the Virgin Island Territories and other U.S. coral ecosystems. The products from this effort provide an accurate assessment of the abundance and distribution of marine habitats surrounding St. John to support more effective management and conservation of ocean resources within the National Park system. This report documents the entire process of benthic habitat mapping in St. John. Chapter 1 provides a description of the benthic habitat classification scheme used to categorize the different habitats existing in the nearshore environment. Chapter 2 describes the steps required to create a benthic habitat map from visual interpretation of remotely sensed imagery. Chapter 3 details the process of accuracy assessment and reports on the thematic accuracy of the final maps. Finally, Chapter 4 is a summary of the basic map content and compares the new maps to a previous NOAA effort. Benthic habitat maps of the nearshore marine environment of St. John, U.S. Virgin Islands were created by visual interpretation of remotely sensed imagery. Overhead imagery, including color orthophotography and IKONOS satellite imagery, proved to be an excellent source from which to visually interpret the location, extent and attributes of marine habitats. NOAA scientists were able to accurately and reliably delineate the boundaries of features on digital imagery using a Geographic Information System (GIS) and fi eld investigations. The St. John habitat classification scheme defined benthic communities on the basis of four primary coral reef ecosystem attributes: 1) broad geographic zone, 2) geomorphological structure type, 3) dominant biological cover, and 4) degree of live coral cover. Every feature in the benthic habitat map was assigned a designation at each level of the scheme. The ability to apply any component of this scheme was dependent on being able to identify and delineate a given feature in remotely sensed imagery.

A point for comparison: the Flower Garden Banks National Marine Sanctuary

A reoccurring goal listed during the creation of Marine Protected Areas (MPAs) is to return the region to a former state. However, limited data is available that describes or characterizes this former condition. Data collected from ecosystems with comparatively limited anthropogenic impacts, can provide invaluable information in suggesting what former states may have looked like. One example is the Flower Garden Banks National Marine Sanctuary which is located 180 kilometers off the coast of Texas. These relatively isolated and pristine banks are capped by substantial scleractinian coral communities, forming excellent habitat for over 200 species of fish. While fishing is permitted, it is limited by difficulty of access. In 2006, NOAA’s Biogeography Branch, in collaboration with the Sanctuary, initiated the first quantitative assessment of fish resources throughout the diveable portions of the Sanctuary. The sampling design and methodologies employed were identical to those that the Branch has utilized in other more impacted regions of the US Caribbean. Initial analyses reveal that fish density and species richness at the Sanctuary were almost two times greater than that found within the US Caribbean and biomass was approximately six times higher. This was due in large part to the presence of sizeable piscivores of the genera Mycteroperca and Dermatolepis. The Sanctuary is one of few minimally impacted locations remaining within the Tropical Western Atlantic. As such, these findings should be considered when attempting to establish a former state or evaluate effectiveness of an MPA in meeting its management goals.

Status of coral reef ecosystems in a marine managed area in St. Croix, USVI [poster]

This poster presents information on the status and trends of coral reef ecosystems in St. Croix, US Virgin Islands (USVI). Data were collected by NOAA’s Center for Coastal Monitoring and Assessment Biogeography Branch (CCMA-BB) from 2001-2006 at 1,275 random locations in and around Buck Island Reef National Monument (BIRNM). The main objective was to quantify changes in fish species and assemblage diversity, abundance, biomass and size structure; to provide spatially explicit information on the distribution of key species or groups of species; and to compare community structure inside versus outside of BIRNM.

Socioeconomic assessment of shrimp farming in relation to local livelihoods in the south-west coastal Bangladesh

Nine different categories of stakeholders in shrimp farming industry ·were assessed to show the socioeconomic impact of shrimp farming in south-west Bangladesh. Among all the stakeholders the shrimp farmer's average own land was 4 ha whereas the seed collectors and faria's had lowest amount of average land, 0.1 and 0.5 ha respectively. The shrimp farming positively impacted to the livelihood of stakeholders. Income of the coastal people, sanitation, working facilities of women, employment, health condition and the literacy rate increased due to shrimp farming. On the other hand shrimp farming had negative impact on the rice production, livestock, drinking water supply, and social conflict and violence had increased due to shrimp farming. There were internal conflicts between different stakeholders; the farias conflict with the depot owners and shrimp farmers, marginal farmers' conflict with the rich shrimp farmers about leasing lands and saline water control, the rice farmers conflicts with the shrimp farmers about agricultural crop production.

Assessment of nutritional quality of 'Shidal' a fermented fish product of northeast India

Shidal is a salt-free fermented fish product prepared from Puntius ss. caught in late monsoon period. Shidal is very popular amongst the inhabitants of Northeast India. The fermentation process of this product takes four to six months in anaerobic condition till the product gains a characteristic flavour and colour. Detailed studies on the biochemical and nutritive qualities of this product are very few. Therefore, in this paper we report the results of the proximate analysis, amino acid and fatty acid compositions. The results suggest that Shidal is a rich source of amino acids as well as of essential fatty acids like linoleic and linolenic acids. The n- 3/n-6 ratio was found 0.51.

Assessment of impact of ten days fishing ban in the major spawning grounds of hilsa (Tenualosa ilisha, Fisher and Bianchi, 1984)

A study was undertaken to find out the impact of ten days fishing ban in the major spawning grounds of hilsa during October to December. The study revealed a positive impact of fishing ban during spawning season on reproduction success of hilsa. Proportionate distribution of male and female hilsa also showed a significant level of distribution in and around the spawning grounds where the ratio was 35:65. Percent of oozing hilsa during fishing ban period in the spawning grounds was observed 1.61%. About 5% spent hilsa was observed in the fish landing centers and was compared with the data of Global Environment Facility (GEF) and Bangladesh Fisheries Research Institute (BFRI) studies and was found about 2.80-3.57 times higher than that of the findings of 2002 and 2003. Estimated egg production value showed about 46,800 Kg of eggs could have been produced that indicating a positive impact of 10 days fishing ban in spawning season. Abundance of higher percentages of gravid hilsa was found which were not available in the same quantity and condition in the non-fishing ban period. During the present investigation, fairly higher amount of spent hilsa and juveniles were also observed in the spawning grounds. Along with the jatka fry, spawn and fries of other fishes were also found in higher quantity than the previous years and thus it is assumed that fishing ban also might have positive impact on the successful breeding of other fishes. Overall, the fishing ban was found effective for successful breeding of hilsa.

Production and quality assessment of fish pickles from mola (Amblypharyngodon mola) fish

Fish pickles (with olive and tamarind) were prepared from mola fish (Amblypharyngodon mola) and their nutritional and food quality were assessed. The quality of the pickle prepared with olive was excellent and the pickle prepared with tamarind was found good. Moisture content of the two pickle products were 43.85% (with tamarind) and 50.89% (with olive). The protein and lipid contents of tamarind added pickle were 19.13 and 35.64% respectively; pickle with olive contained less protein (13.16%) compared to tamarind added mola pickle. Lipid contents were almost same in both cases. Ash content of two pickles was also found similar (1.00%). The quality of mola pickles stored either in cool condition (4°C) with vinegar or at room temperature with Na-benzoate were found good for consumption up to 90 days of storage. All of the fish pickles preserved under different condition were found in acceptable condition up to 240 days storage and pickle with vinegar stored at 4°C was found good for consumption at the end of 240 days.

Transboundary diagnostic analysis. Vol. 2. Background and environmental assessment

The Transboundary Diagnosis Analysis(TDA) quantifies and ranks water-related environmental transboundary issues and their causes according to the severity of environmental and/or socio-economic impacts. The three main issues in BOBLME are; overexploitation of marine living resources; degradation of mangroves, coral reefs and seagrasses; pollution and water quality. Volume 2 contains background material that sets out the bio-physical and socio-economic characteristics of the BOBLME; an analysis of the legal,policy and administrative context; and an assessment of the status of marine living resources and marine environment in coastal areas

Population dynamics and stock assessment of the common carp (Cyprinus carpio) in Iranian waters of the Caspian Sea

In order to come up with the responsible fishing pattern of common carp, there was a need to identify some of the biological characteristics and stock assessment of carp in Iranian waters of the Caspian Sea .The fork length ,weight ,age ,growth parameters of von bertalanffy and mortality rates of common carp were estimated from oct 2006 to sept 2007.Based on the exponential relationship between length and weight in the size range 6.3-65.6 cm ,b was calculated 2.895, 2.843 and 2.925 respectively for combined sexes ,males and females. The mean condition factor was 1.9 which is close to the ideal condition.The results from measuring 3170 specimens ,were showed the first fork length of maturity was 30 cm for males and 32 cm for females. The results indicated that females were predominate and sex ratio was 0.66:1 (M:F) and chi-squares analysis showeda significant difference between males and females.(p<0.05).Length infinity and growth coefficient were calculated by three different methods as below: Length frequency analysis : k=0.17 L∞ =68.04 Age-Length Key k=0.15 L∞ =74.25 Back calculation : k=0.14 L∞ =68.4 The mortality parameters and exploitation rate were estimated as below : Z=0.73 per year M=0.31 per year F=0.42 per year E=0.56 Refer to amount of common carp catch in 2007 -08 ,biomass was calculated 9640.2 tones by jone's cohort analysis and MSY 2374.5 tones.According to analysis ,the number of common carp in the distribution area (Iranian part of the Caspian Sea ) was estimated 24 millions in the 2006-07.