The soft-bottom macrobenthos of Gray's Reef National Marine Sanctuary and nearby shelf waters off the coast of Georgia, USA

As part of an ongoing ecological assessment of the Gray's Reef National Marine Sanctuary (GRNMS), a 58-km² marine protected area 32 km off the coast of Georgia, USA, surveys of benthic macroinfaunal communities, contaminant levels in sediments and biota, and general habitat conditions were conducted during 2000-2002 at 20 stations within the sanctuary and along three cross-shelf transects in nearby shelf waters. Macroinfaunal community structure and composition exhibited distinct cross-shelf patterns associated with sediment granulometry, depth and possibly other factors related to shoreline proximity (e.g., erosional effects, recruitment of estuarine species). Finer-scale spatial patterns of benthic fauna among stations within the sanctuary appear to be related to proximity to live-bottom habitat and other features of seafloor structure (e.g., rippled vs. flat sand). Population densities of dominant fauna within the sanctuary also varied considerably among years, resulting in shifts in the ranking of dominants at most stations. Chemical contaminants generally were at low background concentrations below probable bioeffect levels and thus are not a likely cause of the observed spatial patterns of benthic fauna. However, trace concentrations of pesticides, PCBs, and PAHs were detectable in sediments and biota throughout the study area, demonstrating that chemicals originating from human activities are capable of reaching the offshore sanctuary environment, possibly from atmospheric deposition or cross-shelf transport of materials outwelled through coastal sounds. Highly diverse infaunal assemblages also were observed within the sanctuary and nearby sites of similar depth, suggesting that the sanctuary is an important reservoir of marine biodiversity. Results of this study should be useful in addressing long-term science and management needs of the GRNMS and in furthering our understanding of broader ecological patterns and dynamics of the surrounding South Atlantic Bight (SAB) ecosystem.     

Characterization of macrofaunal assemblages associated with sponges and tunicates collected off the southeastern United States

Abstract. Sponges can serve as hosts to invertebrate assemblages that live and reproduce within them. Sponges also constitute a major part of the benthic epifaunal community on the continental shelf of the southeastern United States; however, little is known about these sponges and the assemblages they harbor. In this study, the associated fauna from a variety of sponges and one species of tunicate collected by submersible from the continental shelf and slope of the southeastern United States at depths in the range 18–875 m were examined. Seventeen sponges, comprising eight species (Ircinia campana, Topsentia sp., Geodia sp., Characella sp., Erylus sp., Apylsina archeri, Cliona sp., and Pheronema carpenteri), and three tunicate colonies (Didemnidae) were fully dissected and all associated organisms were identified and counted. Additionally, the sponges Pheronema annae (951 m) and P. carpenteri (770 m) represent new records for the region. The diversity (H′) and density of associates varied considerably among hosts; the densities of associates ranged 0.4–11,684 per 1 L of host volume. Polychaete worms were the most common organisms found, with one species, Haplosyllis spongicola, being especially abundant in I. campana, Topsentia sp., and Cliona sp. The amphipods Ericthonius punctatus and Leucothoe cf. spinicarpa, as well as decapods such as snapping shrimp (Synalpheus sp.) and crabs (e.g., Pilumnus floridana, Micropanope urinator), were also common. The number of symbiont taxa did not significantly increase as the sponge size increased. However, weak positive trends were found between the diversity of associates and increasing canal diameter. Sponges and tunicates were judged to represent legitimate ecological communities harboring a complete food web as well as gravid and juvenile individuals.     

Harnessing Senescence Delaying Gases Nitric Oxide and Nitrous Oxide: A Novel Approach to Postharvest Control of Fresh Horticultural Produce

The effects of ethylene on postharvest horticultural produce are a major limiting factor in their orderly and efficient marketing. Nitric oxide and nitrous oxide have recently been shown to inhibit ethylene action and synthesis in higher plants and their potential for commercial use to extend the storage and marketing life of fruits, vegetables and flowers is reviewed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Origin of the Sulawesi macaques (Cercopithecidae: Macaco) as suggested by mitochondrial DNA phylogeny

One of the sharpest biogeographical transitions in the world occurs between the Indonesian islands of Borneo and Sulawesi; this transition is demarcated by Wallace's line. Macaque monkeys represent an interesting anomaly to faunal distributions in this region as they occur on both sides of Wallace's line, with Macacafascicularis, M. nemestrina and other species to the west and seven Sulawesi species to the east. We have investigated macaque evolution and dispersal in the Sunda region and Sulawesi using phylogenetic analysis of mitochondrial DNA sequences. Female philopatry of macaques, which causes sharp geographic clustering of maternally inherited mitochondrial DNA haplotypes, makes mitochondrial phytogenies particularly useful for investigating ancient patterns of dispersal. Results of this study suggest the following: (1) M. fascicularis is not a sister taxon to any species of Sulawesi macaque; (2) haplotypes of some M. nemestrina have a sister relationship to northern and central Sulawesi macaques, while haplotypes of other M. nemestrina have a sister relationship to soudiern Sulawesi macaques; (3) Sulawesi was probably colonized by macaques twice, once to the base of the northern peninsula now occupied by M. hecki and once to the southwestern peninsula now occupied by M, mama; and (4) within north/central and southern Sulawesi, patterns of dispersal are largely consistent with contemporary and past geography of the island, with the exception of a geographically discontinuous relationship between M. nigra and a portion of M. tonkeana from a region in northwest central Sulawesi.     

Granular Formulations of Steinernema carpocapsae (strain All) (Nematoda: Rhabditida) with Improved Shelf Life

Shelf life (nematode survival) of Steinernema carpocapsae (strain All) nematodes at 21 C in "Pesta" granules, made by a pasta-like process, was increased from 8 to 26 weeks by incorporating low concentrations of formaldehyde. Pesta samples containing an average of 427,000 nematodes/g were prepared with wheat flour (semolina or bread flour), kaolin, bentonite, peat moss, nematode slurry, and formaldehyde (0-1.4% w/w) and were dried to a water content of 23.6-26.9%. Nematodes emerged from Pesta (S. carpocapsae) granules when placed in water or on moist filter paper. Incorporation of 0.2% w/w formaldehyde (nominal; 0.05% by analysis) was optimum for increasing nematode survival in semolina-based Pesta, and also inhibited fungal growth on the granules. Bread flour Pesta samples prepared by formaldehyde addition to the nematode slurry prior to dough preparation, rather than by addition to a mixture of dry ingredients, had longer shelf life. Nematodes recovered from granules made with 0.2% formaldehyde and stored 20 weeks at 21 C caused 100% mortality of wax moth (Galleria mellonella) larvae.     

Cationic polymer precipitation for enhanced impurity removal in downstream processing

Precipitation can be used for the removal of impurities early in the downstream purification process of biologics, with the soluble product remaining in the filtrate through microfiltration. The objective of this study was to examine the use of polyallylamine (PAA) precipitation to increase the purity of product via higher host cell protein removal to enhance polysorbate excipient stability to enable a longer shelf life. Experiments were performed using three monoclonal antibodies (mAbs) with different properties of isoelectric point and IgG subclass. High throughput workflows were established to quickly screen precipitation conditions as a function of pH, conductivity and PAA concentrations. Process analytical tools (PATs) were used to evaluate the size distribution of particles and inform the optimal precipitation condition. Minimal pressure increase was observed during depth filtration of the precipitates. The precipitation was scaled up to 20L size and the extensive characterization of precipitated samples after protein A chromatography showed >75% reduction of host cell protein (HCP) concentrations (by ELISA), >90% reduction of number of HCP species (by mass spectrometry), and >99.8% reduction of DNA. The stability of polysorbate containing formulation buffers for all three mAbs in the protein A purified intermediates was improved at least 25% after PAA precipitation. Mass spectrometry was used to obtain additional understanding of the interaction between PAA and HCPs with different properties. Minimal impact on product quality and <5% yield loss after precipitation were observed while the residual PAA was <9 ppm. These results expand the toolbox in downstream purification to solve HCP clearance issues for programs with purification challenges, while also providing important insights into the integration of precipitation–depth filtration and the current platform process for the purification of biologics.     

Extremotrophs, extremophiles and broadband pigmentation strategies in a high arctic ice shelf ecosystem

Remnant ice shelves along the northern coast of Ellesmere Island, Nunavut, Canada ( approximately 83 degrees N) provide a habitat for cryo-tolerant microbial mat communities. Bioassays of bacterial and primary production were undertaken to quantify the short-term physiological response of the mats to changes in key variables that characterize this cryo-ecosystem (salinity, irradiance and temperature). The heterotrophic versus autotrophic community responses to these stressors differed markedly. The heterotrophic bacteria were extremophilic and specifically adapted to ambient conditions on the ice shelf, whereas the autotrophic community had broader tolerance ranges and optima outside the ambient range. This latter, extremotrophic response may be partly due to a diverse suite of pigments including oligosaccharide mycosporine-like amino acids, scytonemins, carotenoids, phycobiliproteins and chlorophylls that absorb from the near UV-B to red wavelengths. These pigments provide a comprehensive broadband strategy for coping with the multiple stressors of high irradiance, variable salinity and low temperatures in this extreme cryo-environment.     

Coastal States in the South China Sea and Submissions on the Outer Limits of the Continental Shelf

The purpose of this article is to examine the outer continental shelf submissions made by the coastal states of the South China Sea and their potential impact on legal and political developments in the South China Sea. In accordance with the United Nations Convention on the Law of the Sea of 1982 and the guidelines of the Commission on the Limits of the Continental Shelf, coastal states are to establish the outer limits of their continental shelf where it extends beyond 200 nautical miles. Meeting this obligation is complicated in enclosed or semienclosed seas where there are maritime disputes such as in the South China Sea.     

Managing Adjacency: Some Legal Aspects of the Relationship Between the Extended Continental Shelf and the International Seabed Area

The Third United Nations Conference on the Law of the Sea (1973–1982) struck a difficult compromise between the definition of the outer limits of the extended continental shelf (ECS) in relation to the international seabed area (the Area) and the making of payments and contributions by the coastal state in relation to production activities on its ECS in Article 82. The implementation of Article 82 underscores a broader and more far-reaching relationship between the continental shelf, and the ECS in particular, and the Area. In some regions there may be a relationship between the exclusive economic zone and the Area, where there is no ECS. Effectively, the relationship translates into realities and expectations of good neighborliness. This article examines this relationship and the possible approaches for the management of identified challenges.     

The Rejection of a Theoretical Beauty: The Foot of the Continental Slope in Maritime Boundary Delimitations Beyond 200 Nautical Miles

This article addresses maritime boundary delimitation concerning the continental shelf beyond 200 nautical miles. The focal point is how the foot of the continental slope can be used as the point of departure in drawing the provisional equidistance line in outer continental shelf boundary delimitations between neighboring states. The article examines the strength and weaknesses of this approach and asks whether the International Tribunal for the Law of the Sea indirectly rejected this approach in the 2012 Bangladesh v. Myanmar Case.