Manual for starch gel electrophoresis: A method for the detection of genetic variation

The procedure to conduct horizontal starch gel electrophoresis on enzymes is described in detail. Areas covered are (I) collection and storage of specimens, (2) preparation of tissues, (3) preparation of a starch gel, (4) application of enzyme extracts to a gel, (5) setting up a gel for electrophoresis, (6) slicing a gel, and (7) staining a gel. Recipes are also included for 47 enzyme stains and 3 selected gel buffers. (PDF file contains 26 pages.)

Technologies and Methodologies for the Detection of Harmful Algae and Their Toxins, St Petersburg, Florida, Oct 22-24 2008 : workshop proceedings

The Alliance for Coastal Technologies (ACT) Workshop "Technologies and Methodologies for the Detection of Harmful Algae and their Toxins" convened in St. Petersburg, Florida, October 22- 24, 2008 and was co-sponsored by ACT (http://act-us.info); the Cooperative Institute for Coastal and Estuarine Environmental Technology (CICEET, http://ciceet.unh.edu); and the Florida Fish and Wildlife Conservation Commission (FWC, http://www.myfwc.com). Participants from various sectors, including researchers, coastal decision makers, and technology vendors, collaborated to exchange information and build consensus. They focused on the status of currently available detection technologies and methodologies for harmful algae (HA) and their toxins, provided direction for developing operational use of existing technology, and addressed requirements for future technology developments in this area. Harmful algal blooms (HABs) in marine and freshwater systems are increasingly common worldwide and are known to cause extensive ecological, economic, and human health problems. In US waters, HABs are encountered in a growing number of locations and are also increasing in duration and severity. This expansion in HABs has led to elevated incidences of poisonous seafood, toxin-contaminated drinking water, mortality of fish and other animals dependent upon aquatic resources (including protected species), public health and economic impacts in coastal and lakeside communities, losses to aquaculture enterprises, and long-term aquatic ecosystem changes. This meeting represented the fourth ACT sponsored workshop that has addressed technology developments for improved monitoring of water-born pathogens and HA species in some form. A primary motivation was to assess the need and community support for an ACT-led Performance Demonstration of Harmful Algae Detection Technologies and Methodologies in order to facilitate their integration into regional ocean observing systems operations. The workshop focused on the identification of region-specific monitoring needs and available technologies and methodologies for detection/quantification of harmful algal species and their toxins along the US marine and freshwater coasts. To address this critical environmental issue, several technologies and methodologies have been, or are being, developed to detect and quantify various harmful algae and their associated toxins in coastal marine and freshwater environments. There are many challenges to nationwide adoption of HAB detection as part of a core monitoring infrastructure: the geographic uniqueness of primary algal species of concern around the country, the variety of HAB impacts, and the need for a clear vision of the operational requirements for monitoring the various species. Nonetheless, it was a consensus of the workshop participants that ACT should support the development of HA detection technology performance demonstrations but that these would need to be tuned regionally to algal species and toxins of concern in order to promote the adoption of state of the art technologies into HAR monitoring networks. [PDF contains 36 pages]

Towed vehicles: undulating platforms as tools for maping coastal processes and water quality assessment, Seaside, California, February 5-7,2007. Workshop proceedings

The Alliance for Coastal Technologies (ACT) Workshop on Towed Vehicles: Undulating Platforms As Tools for Mapping Coastal Processes and Water Quality Assessment was convened February 5-7,2007 at The Embassy Suites Hotel, Seaside, California and sponsored by the ACT-Pacific Coast partnership at the Moss Landing Marine Laboratories (MLML). The TUV workshop was co-chaired by Richard Burt (Chelsea Technology Group) and Stewart Lamerdin (MLML Marine Operations). Invited participants were selected to provide a uniform representation of the academic researchers, private sector product developers, and existing and potential data product users from the resource management community to enable development of broad consensus opinions on the application of TUV platforms in coastal resource assessment and management. The workshop was organized to address recognized limitations of point-based monitoring programs, which, while providing valuable data, are incapable of describing the spatial heterogeneity and the extent of features distributed in the bulk solution. This is particularly true as surveys approach the coastal zone where tidal and estuarine influences result in spatially and temporally heterogeneous water masses and entrained biological components. Aerial or satellite based remote sensing can provide an assessment of the aerial extent of plumes and blooms, yet provide no information regarding the third dimension of these features. Towed vehicles offer a cost-effective solution to this problem by providing platforms, which can sample in the horizontal, vertical, and time-based domains. Towed undulating vehicles (henceforth TUVs) represent useful platforms for event-response characterization. This workshop reviewed the current status of towed vehicle technology focusing on limitations of depth, data telemetry, instrument power demands, and ship requirements in an attempt to identify means to incorporate such technology more routinely in monitoring and event-response programs. Specifically, the participants were charged to address the following: (1) Summarize the state of the art in TUV technologies; (2) Identify how TUV platforms are used and how they can assist coastal managers in fulfilling their regulatory and management responsibilities; (3) Identify barriers and challenges to the application of TUV technologies in management and research activities, and (4) Recommend a series of community actions to overcome identified barriers and challenges. A series of plenary presentation were provided to enhance subsequent breakout discussions by the participants. Dave Nelson (University of Rhode Island) provided extensive summaries and real-world assessment of the operational features of a variety of TUV platforms available in the UNOLs scientific fleet. Dr. Burke Hales (Oregon State University) described the modification of TUV to provide a novel sampling platform for high resolution mapping of chemical distributions in near real time. Dr. Sonia Batten (Sir Alister Hardy Foundation for Ocean Sciences) provided an overview on the deployment of specialized towed vehicles equipped with rugged continuous plankton recorders on ships of opportunity to obtain long-term, basin wide surveys of zooplankton community structure, enhancing our understanding of trends in secondary production in the upper ocean. [PDF contains 32 pages]

Biological Platforms for Environmental Sensor technologies and their uses as indicators of the marine environment, Seward, Alaska, September 19-21, 2007: workshop proceedings

The Alliance for Coastal Technologies (ACT) Workshop entitled, "Biological Platforms as Sensor Technologies and their Use as Indicators for the Marine Environment" was held in Seward, Alaska, September 19 - 21,2007. The workshop was co-hosted by the University of Alaska Fairbanks (UAF) and the Alaska SeaLife Center (ASLC). The workshop was attended by 25 participants representing a wide range of research scientists, managers, and manufacturers who develop and deploy sensory equipment using aquatic vertebrates as the mode of transport. Eight recommendations were made by participants at the conclusion of the workshop and are presented here without prioritization: 1. Encourage research toward development of energy scavenging devices of suitable sizes for use in remote sensing packages attached to marine animals. 2. Encourage funding sources for development of new sensor technologies and animal-borne tags. 3. Develop animal-borne environmental sensor platforms that offer more combined systems and improved data recovery methodologies, and expand the geographic scope of complementary fixed sensor arrays. 4. Engage the oceanographic community by: a. Offering a mini workshop at an AGU ocean sciences conference for people interested in developing an ocean carbon program that utilizes animal-borne sensor technology. b. Outreach to chemical oceanographers. 5. Min v2d6.sheepserver.net e and merge technologies from other disciplines that may be applied to marine sensors (e.g. biomedical field). 6. Encourage the NOAA Permitting Office to: a. Make a more predictable, reliable, and consistent permitting system for using animal platforms. b. Establish an evaluation process. c. Adhere to established standards. 7. Promote the expanded use of calibrated hydrophones as part of existing animal platforms. 8. Encourage the Integrated Ocean Observing System (IOOS) to promote animal tracking as effective samplers of the marine environment, and use of animals as ocean sensor technology platforms. [PDF contains 20 pages]

Cultural methods of detection for microorganisms: recent advances and successes

Most microbiological methods require culture to allow organisms to recover or to selectively increase, and target organisms are identified by growth on specific agar media. Many cultural methods take several days to complete and even then the results require confirmation. Alternative techniques include the use of chromogenic and fluorogenic substances to identify bacteria as they are growing, selective capture using antibodies after short periods of growth, molecular techniques, and direct staining with or without flow cytometry for enumeration and identification. Future microbiologists may not use culture but depend on the use of specific probes and sophisticated detection systems.

Detection of specific bacteria in water: implications of survival strategy

It is widely recognised that conventional culture techniques may underestimate true viable bacterial numbers by several orders of magnitude. The basis of this discrepancy is that a culture in or on media of high nutrient concentration is highly selective (either through ”nutrient shock” or failure to provide vital co-factors) and decreases apparent diversity; thus it is unrepresentative of the natural community. In addition, the non-culturable but viable state (NCBV) is a strategy adopted by some bacteria as a response to environmental stress. The basis for the non-culturable state is that cells placed in conditions present in the environment cannot be recultured but can be shown to maintain their viability. Consequently, these cells would not be detected by standard water quality techniques that are based on culture. In the case of pathogens, it may explain outbreaks of disease in populations that have not come into contact with the pathogen. However, the NCBV state is difficult to attribute, due to the failure to distinguish between NCBV and non-viable cells. This article will describe experiences with the fish pathogen Aeromonas salmonicida subsp. salmonicida and the application of molecular techniques for its detection and physiological analysis.

Detection of cryptosporidium oocysts in water and environmental concentrates

Whilst current methods for the isolation and enumeration of Cryptosporidium spp. oocysts in water have provided some insight into their occurrence and significance, they are regarded as being inefficient, variable and time-consuming, with much of the interpretation being left to the expertise of the analyst. Two expectations of novel developments are to reduce the variability and subjectivity associated with the isolation and identification of oocysts. Flocculation, immunomagnetisable and flow cytometric techniques, for concentrating oocysts from water samples, should prove more reliable than current methods, whilst the development of more avid and specific monoclonal antibodies in conjunction with the use of nuclear fluorochromes will aid identification. Further insight into the viability, taxonomy, species identification, infectivity and virulence of the parasite should be forthcoming through the use of techniques such as the polymerase chain reaction, in situ hybridisation and non-uniform alternating current electrical fields. Such information is necessary in order to enable microbiologists, epidemiologists, engineers, utility operators and regulators to assess the safety of a water supply, with respect to Cryptosporidium contamination, more effectively.