Enzyme activities in the water column and in shallow permeable sediments from the northeastern Gulf of Mexico

The activities of extracellular enzymes that initiate the microbial remineralization of high molecular weight organic matter were investigated in the water column and sandy surface sediments at two sites in the northeastern Gulf of Mexico. Six fluorescently labeled polysaccharides were hydrolyzed rapidly in the water column as well as in permeable sediments. This result contrasts with previous studies carried out in environments dominated by fine-grained muds, in which the spectrum of enzymes active in the water column is quite limited compared to that of the underlying sediments. Extracts of Spirulina, Isochrysis, and Thalassiosira were also used to measure hydrolysis rates in water from one of the sites. Rates of hydrolysis of the three plankton extracts were comparable to those of the purified polysaccharides. The broad spectrum and rapid rates of hydrolysis observed in the water column at both sites in the northeastern Gulf of Mexico may be due to the permeable nature of the sediments. Fluid flux through the sediments is sufficiently high that the entire 1.5 m deep water column could filter though the sediments on timescales of a few days to two weeks. Movement of water through sediments may also transport dissolved enzymes from the sediment into the water column, enhancing the spectrum as well as the rate of water column enzymatic activities. Such interaction between the sediments and water column would permit water column microbial communities to access high molecular weight substrates that might otherwise remain unavailable as substrates.     

Surface associations of enzymes and of organic matter: Consequences for hydrolytic activity and organic matter remineralization in marine systems

The extent to which marine organic matter is associated with surfaces and the consequences of such associations for organic matter remineralization are the focus of considerable attention. Since extracellular enzymes operating outside microbial cells are required to hydrolyze organic macromolecules to sizes sufficiently small for substrate uptake, the effects of surface interactions–on enzymes as well as on substrates–for hydrolytic activity also require investigation. We used a simplified laboratory system consisting of a free (dissolved) polysaccharide (pullulan) and the same polysaccharide tethered to agarose beads to restrict mobility, plus the corresponding free enzyme (pullulanase) and the same enzyme sorbed to montmorillonite (Mte), to investigate systematically the consequences of surface associations of enzymes and of substrates on hydrolytic activity. Changes in substrate molecular weight were monitored with time to measure the course of enzymatic hydrolysis. Although hydrolysis of free substrate was nearly complete after 2 min incubation with the free enzymes, the sorbed enzymes also effectively hydrolyzed free substrate, and the data suggest that they retained activity longer in solution compared to the free enzymes. Sorbed enzymes progressively hydrolyzed the free substrate from > 50 kD to lower molecular weights during a 24 h incubation, with a final product distribution on average showing only 1.4% and 10.3% of substrate still in the > 50 kD and 10 kD size classes, while 46.6%, 29.3%, and 12.5% of substrate was in the 4 kD, monomer, and free tag size classes, respectively. This product distribution was very similar to that of the free substrate/free enzyme experiment. Tethering the substrate to agarose beads led to lower substrate release (2–3% of total substrate after 98 h incubation) into solution compared to the free substrate case. For tethered substrates, the state of the enzyme (free or sorbed) measurably affected the molecular weight distribution of the hydrolysis products, with free enzymes producing a higher fraction of high molecular weight hydrolysis products (28.7 ± 5.4% of substrate > 50 kD at the end of the incubation) compared to sorbed enzymes (11.6 ± 2.8% of substrate > 50 kD at the end of the incubation.) Tethered substrates were also hydrolyzed in a sediment slurry from surface sediments from Cape Lookout Bight, North Carolina; 0.1% of total substrate was released by enzymes naturally present in 1 cm³ of sediment after 144 h incubation, demonstrating that the enzymes naturally present in marine sediments are also capable of accessing tethered substrates. These investigations suggest that surface associations of enzymes in marine systems may extend the active lifetime of such enzymes, providing an opportunity for hydrolysis over longer periods of time and producing a different size spectrum of hydrolysis products relative to free enzymes. Furthermore, in well-mixed systems, surface-associated enzymes can hydrolyze substrates whose mobility is restricted, highlighting the importance of processes such as resuspension and bioturbation on organic matter remineralization.     

Application of fluorescence spectroscopic techniques and probes to the detection of biopolymer degradation in natural environments

The activities and substrate specificities of extracellular enzymes in natural systems are not well understood, despite their critical role in microbial remineralization of organic carbon. These enzymes initiate organic carbon degradation by selectively hydrolyzing high molecular weight substrates to lower molecular weight products which can be transported into cells. A set of single- and dual-labeled fluorescent polysaccharides was synthesized and characterized to explore a variety of approaches for measuring enzymatic hydrolysis of biopolymers via photophysical techniques, focusing particularly on rapid and robust optical techniques which are amenable to field measurements in remote locales. A shotgun-labeling approach yielded dual-labeled probes that exhibited substantial donor fluorophore quenching. The photophysical response of these probes to hydrolysis via purified enzymes was investigated in the lab, and fluorescence polarization proved to be a rapid and reliable technique for monitoring probe hydrolysis. Initial field results were also obtained from hydrolysis experiments in sediment porewaters. Because polarization measurements are rapid and simple, this approach could be used to follow the extracellular enzymatic hydrolysis of a wide range of biopolymers which fuel microbial metabolism.     

Electron paramagnetic resonance spectroscopy as a novel approach to measure macromolecule–surface interactions and activities of extracellular enzymes

The dynamics of high molecular weight organic matter in marine systems are influenced by molecular conformation, interactions with surfaces and susceptibility to enzymatic hydrolysis, parameters that are difficult to observe experimentally. Here we use electron paramagnetic resonance spectroscopy (EPR) and spin-labeled (SL-) polysaccharides to monitor the sorption of SL-polysaccharides to natural sediment surfaces and to montmorillonite and to observe decreases in polysaccharide size due to enzymatic hydrolysis. SL-pullulan, SL-xylan and SL-maltoheptaose all sorbed rapidly to muddy sediments but not to sandy sediments. SL-pullulan and SL-maltoheptaose also both sorbed to montmorillonite; however, SL-pullulan reached substantially greater final surface loadings than did SL-maltoheptaose. Using EPR has the advantages of being rapid (spectra can be acquired in 100 seconds), non-destructive and functional in complex media, including sediment slurries, muddy water or other optically opaque samples, permitting investigation of the interactions between biomacromolecules, extracellular enzymes and mineral surfaces in aquatic environments.     

Fate and recycling of carbon compounds

This article is a brief and selective introduction to the literature and concepts concerning the fate and recycling of carbon compounds in the marine environment. It provides a framework for the other papers in the session and emphasises the areas of ignorance and the implications to be drawn from them.The fate of carbon compounds in oceanic water columns and sediments is reviewed in terms of regions and processes. Particular attention is given to certain regions — surface film and associated zone, the water column and the upper layer of the bottom sediments — and processes — microbial activity, association of organic materials and minerals and the formation and diagenesis of particulate organic matter.Most of the organic matter in the ocean is rapidly recycled but the processes, rates and fluxes are poorly understood. Major areas of ignorance include the half-lives of individual compounds and of classes of compounds and the role of microorganisms, both in the water column and in the bottom sediments. Measurements and experiments need to be conducted in the oceans and in the laboratory. Chemical and biochemical changes in the short and long term require recognition if the residual organic matter of sediments is to be interpreted in terms of past oceanic conditions.     

南海沉积物细菌胞外蛋白酶在家族水平上的多样性

海洋产蛋白酶细菌及其分泌的胞外蛋白酶在降解有机氮以推动海洋氮循环进行方面发挥着重要作用,但目前对这二者多样性的认识非常有限。本研究自南海沉积物中筛选得到90株产蛋白酶细菌,并通过N-端氨基酸序列,分析了其胞外蛋白酶在家族水平上的多样性。16S rRNA基因序列分析的结果表明,筛选的产蛋白酶细菌均属于Gammaproteobacteria纲,且大多数属于Alteromonadales目和Vibrionales目的不同属。对其中14株菌株的14个胞外蛋白酶的N-端氨基酸序列分析表明,所有这些蛋白酶属于金属蛋白酶的M4家族或丝氨酸蛋白酶的S8家族。本研究提供了海洋沉积物产蛋白酶细菌在类群及其胞外蛋白酶在类型上的新细节,这将有助于全面了解微生物酶促降解海洋沉积物有机氮的过程和机理。     

胶州湾沉积物中氨基酸对有机质降解及细菌源贡献的指示作用解析

基于对胶州湾表层沉积物中总可水解氨基酸（THAA）的含量、组成、构型及分布特征的系统研究,通过氨基酸碳氮归一化产率（THAA-C%,THAA-N%）、降解因子DI、反应活性指数RI以及D型氨基酸占比（D-AA%,摩尔百分比）等指标结合碳氮比（TOC/TN）、碳稳定同位素（δ13C）探析了胶州湾沉积物中有机质的来源与降解状态,利用细菌源有机质及胞外肽酶活性（EEA）探讨了微生物在有机质迁移转化过程中的作用与贡献。结果表明,胶州湾表层沉积物中氨基酸平均含量为（7.60±3.64）μmol/g,在陆源与海源混合影响下,其水平分布呈现湾内高于湾外、湾内东部高于西部的特点,表明湾内东部陆源输入对沉积物THAA具有较高贡献。THAA-C%、THAA-N%、DI、RI以及D-AA%等指示因子均显示胶州湾表层沉积物中有机质的降解程度呈现湾外高于湾内、湾内东部高于西部的变化趋势,有机质来源、微生物活性与上覆水水深共同影响了有机质的降解程度。胶州湾表层沉积物中细菌源有机碳的贡献率为（29.35±18.73）%,其水平分布显示出湾内西部与湾外相近且高于湾内东部的特点。细菌胞外肽酶活性（EEA）平均为（0.81±1.31）nmol/（g·h）（以MCA计）,整体分布趋势与细菌贡献率相反,呈现湾内东部高于湾内西部和湾外的特性。沉积物中有机质的不同海源、陆源占比决定了有机质的可降解性,而有机质的降解程度进一步影响了细菌源有机质的贡献与胞外肽酶活性。     

大河影响下的陆架边缘海沉积有机碳的再矿化作用

大河影响下的陆架边缘海(RiOMars)是陆源有机碳的主要沉积汇,是陆海相互作用最重要的区域,在全球碳的生物地球化学循环中发挥着重要作用。受到RiOMars系统内频繁的物理和生物等改造作用的影响,该区沉积的有机碳并没有得到很好地保存而被永久埋藏,而是发生了显著的再矿化分解。本文首先对目前常用的基于O2消耗速率和CO2产生速率的两类测定RiOMars系统沉积有机碳再矿化速率的方法进行了介绍,分析了各自的优缺点和适用性,进而从碳的形态转化、表层沉积物混合均匀、形成次氧化的氧化还原条件、有机碳保存效率低、发生反风化作用和微生物发挥着重要作用等几个方面对RiOMars系统沉积物发生再矿化作用时的主要过程和特征进行了剖析,以期深入认识边缘海的再矿化作用及其对边缘海碳汇的影响。     

The oxygen minimum zone of the eastern South Pacific

In spite of the fact that oxygen-deficient waters with ⩽20 μM of dissolved oxygen—known as oxygen minimum zones (OMZs)—occupy only ∼1% of the volume of the global ocean, they disproportionately affect global biogeochemical cycles, particularly the nitrogen cycle. The macrobiota diversity in OMZs is low, but the fauna that do inhabit these regions present special adaptations to the low-oxygen conditions. Conversely, microbial communities in the OMZ water column and sediments are abundant and phylogenetically and metabolically very diverse, and microbial processes occurring therein (e.g., denitrification, anammox, and organic matter degradation) are important for global marine biogeochemical cycles. In this introductory article, we present the collection of papers for the special volume on the OMZ of the eastern South Pacific, one of the three main open-ocean oxygen-deficient regions of the global ocean. These papers deal with aspects of regional oceanography, inorganic and organic geochemistry, ecology, and the biochemistry of micro and macro organisms—both in the plankton and in the sediments—and past changes in the fish scales preserved in the sediments bathed by OMZ waters.     

The mineralization of chitin in the sediments of the Ythan estuary, aberdeenshire, Scotland

Reliable methods have been developed for the quantitative estimation of chitin, chitosan and chitin deacetylase in sediment samples, as well as two methods for the estimation of chitinase in these samples. The differing substrates used in the latter two methods have produced distinct ‘depth profiles’ for chitinase activity which suggest the presence of more than one chitinolytic system in the sediments of the Ythan estuary. Studies on the mineralization of chitin in these sediments have revealed a complex system with a number of unexpected features. The rate of chitin degradation was reduced in late summer, possibly due to the heavy seaweed growth which covers the mud flats at this time, although the potential degradative activities of the enzymes chitinase and chitin deacetylase were higher in summer than in winter. The proportional rates of degradation of chitin, chitosan and cellulose were similar in both the absence and presence of seaweed overgrowth, although the apparent rates of degradation of these substrates were higher in the absence of seaweeds. Core samples have indicated that the major input of chitin to the sediment ecosystem occurs via deposition at the surface, while the input of chitosan (the deacetylated polymer) occurs via its production within the sediment, primarily within the upper 5 cm of sediment. The concentrations of both these polymers decreased sharply below an ‘interface’ between aerobic and anoxic sediments.     

Trace element and molecular markers of organic carbon dynamics along a shelf–basin continuum in sediments of the western Arctic Ocean

Molecular organic biomarkers together with trace element composition were investigated in sediments east of Barrow Canyon in the western Arctic Ocean to determine sources and recycling of organic carbon in a continuum from the shelf to the basin. Algal biomarkers (polyunsaturated and short-chain saturated fatty acids, 24-methylcholesta-5,24(28)-dien-3β-ol, dinosterol) highlight the substantial contribution of organic matter from water column and sea-ice primary productivity in shelf environments, while redox markers such as acid volatile sulfide (AVS), Mn, and Re indicate intense metabolism of this material leading to sediment anoxia. Shelf sediments also receive considerable inputs from terrestrial organic carbon, with biomarker composition suggesting the presence of multiple pools of terrestrial organic matter segregated by age/lability or hydrodynamic sorting. Sedimentary metabolism was not as intense in slope sediments as on the shelf; however, sufficient labile organic matter is present to create suboxic and anoxic conditions, at least intermittently, as organic matter is focused towards the slope. Basin sediments also showed evidence for episodic delivery of labile organic carbon inputs despite the strong physical controls of water depth and sea-ice cover. Principal components analysis of the lipid biomarker data was used to estimate fractions of preserved recalcitrant (of terrestrial origin) and labile (of marine origin) organic matter in the sediments, with ranges of 12–79%, 14–45%, and 37–66% found for the shelf, slope, and basin cores, respectively. On average, the relative preserved terrestrial organic matter in basin sediments was 56%, suggesting exchange of organic carbon between nearshore and basin environments in the western Arctic.     

The supply and preservation of ancient and modern components of organic carbon in the Canadian Beaufort Shelf of the Arctic Ocean

The provenance of organic matter in sediments from the Mackenzie River and Beaufort Shelf was investigated using the stable carbon and radiocarbon isotopic compositions of bulk organic matter and the stable carbon isotopic compositions of individual organic compounds, including lignin-derived phenols and lipid-derived fatty acids. Most river suspended sediments and shelf surface sediments contained organic carbon characterized by highly depleted Δ¹⁴C values that were consistent with average radiocarbon ages exceeding 7000 years. The stable carbon isotopic signatures of lignin phenols were uniformly depleted (−25≥δ¹³C≥−32‰), indicating the predominant contributions of C₃ vascular plant sources. The isotopic compositions of C₁₄ and C₁₆ fatty acids exhibited important contrasts between the river (−36‰ to −40‰) and shelf (−25‰ to −29‰) sediments that were consistent with contributions from freshwater algae and/or vascular plants in the former and marine phytoplankton in the latter. Using ¹⁴C isotopic mass balance, the abundances of modern and ancient organic matter were quantitatively constrained. The fate of organic matter in the Beaufort Shelf was explored by normalizing these abundances to the specific surface area of sediments. Ancient organic carbon, which may include old pre-aged soil material as well as fossil bitumen or kerogen, accounted for the majority (∼70%) of the particulate organic matter exported by the Mackenzie River and deposited in surface sediments of the Beaufort Shelf. Modern organic carbon accounted for ∼30% in both river and shelf sediments, with significant contributions from vascular plant-derived materials in both river and shelf samples and from marine algae in the shelf sediments. Respiration (and/or leaching) of particle-bound marine organic matter dominates the carbon metabolism in the Mackenzie Delta/Beaufort Shelf region. However, land-derived pools, including modern carbon derived from vascular plants as well as ancient carbon also appeared to undergo a degree of post-depositional degradation prior to burial in the shelf. These novel source apportionments are reflected in an updated carbon budget for the study area.     

Vertical Patterns of Suspended Matter Characteristics Along a Coastal-basin Transect in the Western Baltic Sea

Suspended particulate matter samples were collected from the water column, the bottom nepheloid layer and the ‘ fluffy layer ’ from four stations along a coastal-basin transect in the Pomeranian Bight, western Baltic Sea. Sampling was performed nine times between October 1996 and December 1998 for various analyses, including electron probe x-ray micro analysis for detailed mineralogical investigations.Specific vertical patterns of clay mineral distributions were found. Suspended particulate matter (SPM) in the bottom nepheloid layer and the ‘ fluffy layer ’ overlying sediments was enriched in organic carbon and hydrated three layer clay minerals, whereas the non-aggregated SPM was dominated by quartz and biogenic opal. It appears that separation effects operate during aggregation of mineral particles and organic matter in repeated cycles of resuspension and settling. No clear seasonal variations in the composition of the SPM were found, in spite of high spatial and temporal variability of biological and physical variables. The results suggest that preferential incorporation, possibly aided by microbiological colonization, of hydrated three layer silicates into the organic flocs is a process that occurs under a wide range of conditions. Because aggregates sink faster than individual particles, aggregate formation led to a relative enrichment of illite and smectite in the near-bottom layers. Considering the affinity of organic contaminants and heavy metals to organic matter, the selective removal of aggregated organic matter and hydrated three-layer clay minerals from the water column and enhanced transport in the near-bottom fluffy layer may be a natural cleansing mechanism operating in the shallow waters of the bight.     

The solid state speciation of copper in surface water particulates and oceanic sediments

A sequential leaching technique has been used to characterize the solid state speciation of total copper (∑Cu) among a number of operationally defined host fractions in surface seawater particulates from the Atlantic Ocean, a diagenetically active hemipelagic sediment core from the eastern Mediterranean, a turbidite - rich sediment core from the Madeira Abyssal Plain and a series of 79 Atlantic Ocean surface or near surface sediments. Around 50% of the ∑Cu in the surface water particulates is held in organic associations. When the material is deposited at the sediment surface, following its entry into the down-column carbon flux, the ∑Cu undergoes phase transformations as the organic carriers are destroyed. However, some of the organically associated copper (Cu₅) is preserved in the sediments, the amount depending on the diagenetic environment of deposition. The relationship between ∑Cu and organic carbon in an oceanic sediment may be masked, but the partitioning speciation data has shown that good correlations can be found between organic carbon and Cu₅. The concentration of Cu₅ in Atlantic Ocean surface sediments is highest in hemipelagic (diagenetically active) sediments deposited in the marginal regions, and lowest in open-ocean (less diagenetically active) sediments of the Mid-Atlantic Ridge and ridge flanks. The marginal sediments contain an average of 20% of their total Cu in an organic association, with the result that these sediments can act as traps for seawater-derived Cu that would normally be regarded as being ‘reactive’ in the marine environment. To a first approximation, the preservation of Cu₅ in the sediments mimics that of primary production in the overlying waters, and so ‘fingerprints’ the operation of the global ocean carbon flux in oceanic deposits. However, the relationship can be perturbed by the off-shelf transport of organic-rich, Cu₅-containing, turbidites which can result in the transfer and burial of organic copper host fractions in open-ocean oxic environments.     

Origin of sedimentary organic matter in the north-western Adriatic Sea

In order to evaluate the origin and the transformation of organic matter on the shallow shelf of the NW Adriatic Sea, organic carbon, total nitrogen and stable isotope ratios of organic carbon were analysed in riverine suspended matter and sediments as well as in marine suspended and sedimentary organic matter, in marine phytoplankton and zooplankton.The deposition of organic matter is influenced by fine sediment concentration. Surface sediments were characterised by highly variable biogeochemical conditions on the sea floor, whereas sub-surface sediments showed a more homogeneous hypoxic/anoxic environment.Low C_org/N ratio and high organic carbon and nitrogen concentrations in riverine suspended organic matter indicate an important contribution of freshwater phytoplankton within rivers, particularly during low flow regimes, which adds to the marine phyto- and zooplankton at shelf locations.In order to evaluate the importance of terrestrial, riverine and marine sources of OM in shelf sediments, a three end-member mixing model was applied to shelf surface sediments using ¹³C/¹²C values for organic matter and N/C ratios. The model showed an elevated contribution of terrestrial organic substances at intermediate depths (10–15 m), mostly corresponding to an area of coarser grain-size, whereas the riverine and marine organic fractions were mainly accumulating near the coast and offshore, respectively.     

A depocenter of organic matter at 7800 m depth in the SE Pacific Ocean

The Atacama trench, the deepest ecosystem of the southern Pacific Ocean (ca. 8000 m depth) was investigated during the Atacama Trench International Expedition. Sediments, collected at three bathyal stations (1040–1355 m depth) and at a hadal site (7800 m) were analyzed for organic matter quantity and biochemical composition (in terms of phytopigments, proteins, carbohydrates and lipids), bacterial abundance, biomass and carbon production and extracellular enzymatic activities. Functional chlorophyll-a (18.0±0.10 mg m⁻²), phytodetritus (322.2 mg m⁻²) and labile organic carbon (16.9±4.3 g C m⁻²) deposited on surface sediments at hadal depth (7800 m) reached concentrations similar to those encountered in highly productive shallow coastal areas. High values of bacterial C production and aminopeptidase activity were also measured (at in situ temperature and 1 atm). The chemical analyses of the Atacama hadal sediments indicate that this trench behaves as a deep oceanic trap for organic material. We hypothesize that, despite the extreme physical conditions, benthic microbial processes might be accelerated as a result of the organic enrichment.     

Microbial colonisation of artificial and deep‐sea sediments in the Arctic Ocean

Although environmental factors such as grain size and organic carbon content may influence the distribution of microbes in marine sediments, there has been little experimental study of the topic to date. To investigate how those sediment variables affect microbial colonisation under in situ conditions, deep‐sea sediments and artificial sediments (glass beads, sands) were incubated in the Arctic deep sea at 2500 m water depth with or without chitin, one of the most important carbon polymers in marine environments. Microbial abundance, biomass, chitobiase activity and changes in community structure were monitored after 7 days and 1 year. In control sediments without chitin addition, no significant changes in microbial abundance, biomass and activity were observed after 1 year. In the presence of chitin, however, considerable increases in these parameters were recorded in all three sediment types tested. Regardless of chitin addition, natural deep‐sea sediments were always associated with higher values of microbial abundance, biomass and activity compared with artificial sediments. Sediment type was always found to be the most significant factor explaining variation in enzymatic activity and bacterial community structure as compared to the effects of chitin amount, incubation time, and changes in cell number or biomass. Overall, this is the first in situ study that has addressed the effects of multiple factors and their interactions on abundance, biomass, activity and community structure of microbial communities in the deep Arctic Ocean.     

Linking bacterial community structure to carbon fluxes in marine environments

Microbial oceanography is undergoing a dramatic revolution thanks to the rapid development of novel techniques that allow the examination of microbial diversity and functions via molecular methods, including genomic and metagenomic analyses. During the past decade, studies have revealed previously unknown and surprisingly diverse bacterial communities in marine waters. These studies have radically changed our understanding of spatiotemporal patterns in marine bacterial community composition and the distribution of specific genes. However, our knowledge of the role of individual bacterial subgroups in oceanic food webs and biogeochemical cycles remains limited. To embed the internal dynamics of bacterial communities into marine biogeochemistry models, the characteristic parameters of individual bacterial subgroups (i.e., growth, mortality, and utilization of dissolved organic matter) must be determined. Here, we survey the approaches used to assess variation in and factors controlling bacterial communities in marine environments, emphasizing the importance of quantitative studies that examine growth and grazing parameters of bacterial subgroups.     

The need and practice of monitoring, evaluating and adapting marine planning and management—lessons from the Great Barrier Reef

An increasing number of scientists and resource managers recognise that successful marine management approaches, including marine spatial planning (MSP), cannot occur without effective monitoring, evaluation and adaptation. These basic components are necessary to ensure that any marine planning or marine management measures are both effective and efficient. While a number of fundamental principles for marine monitoring, evaluation and adaptive management exist, there are varying levels of understanding about how these should be undertaken and what they may achieve. Challenges include the development of realistic and measurable objectives and indicators against which effectiveness can be practically measured. The matter becomes even more complicated as the focus of marine planning and management strategies changes from ‘single species’ to ‘habitats’ and ‘ecosystems’ that may enable a diversity of permitted uses consistent with a variety of overall objectives. Over the last 30 years, the Great Barrier Reef Marine Park (GBRMP) has successfully established a multiple-use spatial management approach that allows both high levels of environmental protection and a wide range of human activities. Drawing on this unique long-term experience in the GBRMP, this article discusses key aspects of effective monitoring and evaluation, and summarises lessons learned from over two decades of adaptive management.