A novel MODIS algorithm to estimate chlorophyll a concentration in eutrophic turbid lakes

A novel approach was developed to estimate phytoplankton biomass in eutrophic turbid lakes, using MODIS bands designed for land and atmospheric studies. The Baseline Normalized Difference Bloom Index (BNDBI) uses the difference of remote-sensing reflectance (Rrs, sr⁻¹) at 555 nm (band 4) and 645 nm (band 1) after baseline correction using bands at 469 nm and 859 nm: (Rrs′(555) − Rrs′(645))/(Rrs′(555) + Rrs′(645)). BNDBI takes advantage of the Chl-a’s absorption minimum near 572 nm and absorption maximum near 667 nm. Using data from Lake Chaohu, the index showed a strong relationship with Chl-a concentrations in conditions that would normally saturate more sensitive ocean-color sensors. Extensive field measurements were used to calibrate and validate the algorithm with unbiased root-mean-square-error (URMSE) of 47.9% when compared to in situ Rrs data. A reduced sensitivity to atmospheric effects was accomplished by using a baseline correction approach, anchored at 469 nm and 859 nm to correct the radiances at 555 nm and 645 nm. Radiative transfer simulations showed that the algorithm can be applied directly to MODIS Rayleigh-corrected reflectance (Rrc) after adjusting algorithm coefficients (URMSE uncertainty of 56.4% for MODIS Rrc data) for Chl-a concentrations <1000 μg L⁻¹. Comparative analyses showed that the index was resistant to changes in turbidity and organic matter concentrations. Theoretical simulations, image comparisons and spectral analyses demonstrated that the index was robust in a range of complex atmospheric and surface conditions, with different aerosol types, optical thickness (τ_a555), solar/viewing geometry, sun glint and thin clouds. A comparison with other MODIS and MERIS Chl-a algorithms for turbid waters showed that BNDBI provided consistent results with the advantage of using MODIS wavebands that remain unsaturated in high turbidity conditions. The BNDBI opens new possibilities to explore bio-optical dynamics in turbid eutrophic lakes using data from a range of satellite sources.     

Harmful algal bloom (HAB) in the East Sea identified by the Geostationary Ocean Color Imager (GOCI)

Harmful Cochlodinium polykrikoides blooms have frequently appeared and caused fatal harm to aquaculture in Korean coastal waters since 1995. We investigated the applicability of GOCI, the world's first Geostationary Ocean Color Imager, in monitoring the distribution and temporal movement of a harmful algal bloom (HAB) that was discovered in the East Sea near the Korean peninsula in August 2013. We identified the existence of C. polykrikoides at a maximum cell abundance of over 6000 cells/mL and a chlorophyll a concentration of over 400 mg/m³. In areas of C. polykrikoides blooms, GOCI remote sensing reflectance (R_rs) spectra demonstrated the typical radiometric features of a HAB, and from the diurnal variations using GOCI-derived chlorophyll concentration images, we were able to identify the vertical migration of the red tide species. We also found that the formation and propagation of the HAB had relations with cold water mass in the coastal region. GOCI can be effectively applied to the monitoring of short-term and long-term movements of red tides.     

Real-time observation,early warning and forecasting phytoplankton blooms by integrating in situ automated online sondes and hybrid evolutionary algorithms

Phytoplankton bloom is one of the most serious threats to water resource, and remains a global challenge in environmental management. Real-time monitoring and forecasting the dynamics of phytoplankton and early warning the risks are critical steps in an effective environmental management. Automated online sondes have been widely used for in situ real-time monitoring of water quality due to their high reliability and low cost. However, the knowledge of using real-time data from those sondes to forecast phytoplankton blooms has been seldom addressed. Here we present an integrated system for real-time observation, early warning and forecasting of phytoplankton blooms by integrating automated online sondes and the ecological model. Specifically, based on the high-frequency data from automated online sondes in Xiangxi Bay of Three Gorges Reservoir, we successfully developed 1–4 days ahead forecasting models for chlorophyll a (chl a) concentration with hybrid evolutionary algorithms (HEAs). With the predicted concentration of chl a, we achieved a high precision in 1–7 days ahead early warning of good (chl a < 25 μg/L) and eutrophic (chl a 8–25 μg/L) conditions; however only achieved an acceptable precision in 1–2 days ahead early warning of hypertrophic condition (chl a ≥ 25 μg/L). Our study shows that the optimized HEAs achieved an acceptable performance in real-time short-term forecasting and early warning of phytoplankton blooms with the data from the automated in situ sondes. This system provides an efficient way in real-time monitoring and early warning of phytoplankton blooms, and may have a wide application in eutrophication monitoring and management.     

Allelopathic effects of Microcystis aeruginosa on green algae and a diatom: Evidence from exudates addition and co-culturing

Allelopathic interactions among phytoplankton species are regarded as one of the important factors contributing to phytoplankton species competition and succession. The role and extent of allelopathic effects of blooming freshwater cyanobacteria on other phytoplankton species in eutrophied waters, however, are still unknown. We examined the allelopathic effect of Microcystis aeruginosa on two common green algae (Scenedesmus quadricauda, Chlorella pyrenoidosa) and a diatom (Cyclotella meneghiniana) by adding exudates from different growth phases and in co-culture tests. Exudates of M. aeruginosa from the exponential growth phase and the stationary phase significantly inhibited the growth of S. quadricauda, C. pyrenoidosa and C. meneghiniana, whereas those from the decline phase increased their growth. The presence of M. aeruginosa extremely inhibited the growth of all tested species in co-cultures within 24 h. Our results indicate that under the tested environmental conditions (25 °C, light 80 μmol quanta m⁻² s⁻¹, manual shaking twice a day), allelopathic effects of M. aeruginosa on other phytoplankton species can significantly contribute to their competitive success.     

Remote sensing of phytoplankton-macrophyte coexistence in shallow hypereutrophic fluvial lakes

We investigated with remote sensing (APEX images) the coexistence of phytoplankton and macrophytes in three interconnected shallow and hypereutrophic fluvial lakes (Mantua Lakes, Northern Italy). High concentrations of chlorophyll-a, up to 60 mg m^?3, were determined in the open water between well-developed stands of floating-leaved, submerged, and emergent macrophytes. Our data suggest a general inhibition of phytoplankton by macrophytes, evidenced by decreasing chlorophyll-a concentrations in proximity of macrophyte stands. Chlorophyll-a concentrations halved in the proximity of emergent stands (~6 mg m^?3 within 21 m from the stand border) when compared to the outer zones (~13 mg m^?3). Contrasting trends were observed for submerged stands, where concentrations decreased inwards from ~8 to ~3 mg m^?3. Floating leaved stands had a neutral effect, chlorophyll-a being nearly constant in both inner and outer zones. Overall, remotely-sensed data allow evaluation of quantitative and spatially defined interactions of macrophytes and phytoplankton at the whole ecosystem scale.     

Remote sensing of chlorophyll-a concentration for drinking water source using genetic algorithms (GA)-partial least square (PLS) modeling

Accurate estimation of phytoplankton chlorophyll-a (Chl-a) concentration in turbid waters through remote sensing is a challenge due to the optical complexity of water constituents. Reflectance spectra and concurrent water quality parameters of 225 samples across the Shitoukoumen Reservoir, the drinking water resource for Changchun City, were used to retrieve Chl-a concentration with high total suspended matter (TSM) during 2006–2008. A combination of genetic algorithms and partial least square (GA-PLS) model was established for Chl-a retrieval through GA to select sensitive spectral variables and PLS for regression. To compare GA-PLS performances, the widely accepted three-band algorithm was implemented for Chl-a concentration estimation. Both GA-PLS and the three-band algorithm have stable performance for the aggregated dataset (R² = 0.85 and 0.81; RPD = 3.95 and 3.61; relative RMSE = 31.7% and 34.2%), with the GA-PLS model performing marginally better. The temporal transferability of the models was validated with the dataset collected in 2006 and 2007 respectively as independent dataset, showing that GA-PLS outperformed the three-band algorithm. Our result also indicated that relative error [(Chl-a_predicted − Chl-a_measured) / Chl-a_measured] showed good linear relation to TSM: Chl-a ratio (R² = 0.84), which implied that TSM concentration exerted significant impact on the accuracy of Chl-a estimation in this case study. As the results were derived from a large number of samples representing a wide range of spatiotemporal variations of pigment under TSM (3.7–472.8 mg/L) concentration influence, the GA-PLS model has great potential for Chl-a estimation for inland waters with similar backgrounds. Nevertheless, the three-band algorithm also has its own merit considering its simplicity for implementation.     

Influence of bio-optical parameter variability on the reflectance peak position in the red band of algal bloom waters

On the basis of field measurements, the quantitatively different relationships of peak position in the red band of the remote sensing reflectance vs. Chl concentration are found in the bloom waters of the diatom Skeletonema costatum and the dinoflagellate Prorocentrum donghaiense in coastal areas of the East China Sea. Model simulations of remote sensing reflectance, R_rs, accounting for the influence of variations in the bio-optical parameters such as chlorophyll fluorescence quantum efficiency, Φ, and specific absorption coefficient, a_ph^⁎, are carried out to analyze the characteristics of this spectral peak. The strong effect of fluorescence on the magnitude of R_rs results in the inhibition of the shift of the peak to longer wavelengths, increasing Φ enhances this effect. Increasing a_ph^⁎, specifically in the red-wavelength band, causes a sharper shift in the red peak position by decreasing the effect of the fluorescence. The dominant parameter governing the slope of the shift is a_ph^⁎. The analysis indicates that the higher a_ph^⁎ of S. costatum in the red region is primarily responsible for the much higher slope of the peak shift than for that of P. donghaiense. We show that the relationship between the peak position and Chl concentration may be useful for discriminating S. costatum blooms from those due to P. donghaiense, although information about chlorophyll fluorescence quantum efficiency should be included. Finally, we show that using the band ratio R_rs(708 nm)/R_rs(665 nm) instead of Chl in the relationship with peak position can be useful for the practical identification of S. costatum blooms from hyperspectral measurements of remote sensing reflectance.     

Best hyperspectral indices for tracing leaf water status as determined from leaf dehydration experiments

Leaf water status information is highly needed for monitoring plant physiological processes and assessing drought stress. Retrieval of leaf water status based on hyperspectral indices has been shown to be easy and rapid. However, a universal index that is applicable to various plants remains a considerable challenge, primarily due to the limited range of field-measured datasets. In this study, a leaf dehydration experiment was designed to obtain a relatively comprehensive dataset with ranges that are difficult to obtain in field measurements. The relative water content (RWC) and equivalent water thickness (EWT) were chosen as the surrogates of leaf water status. Moreover, five common types of hyperspectral indices including: single reflectance (R), wavelength difference (D), simple ratio (SR), normalized ratio (ND) and double difference (DDn) were applied to determine the best indices. The results indicate that values of original reflectance, reflectance difference and reflectance sensitivity increased significantly, particularly within the 350–700 nm and 1300–2500 nm domains, with a decrease in leaf water. The identified best indices for RWC and EWT, when all the species were considered together, were the first derivative reflectance based ND type index of dND (1415, 1530) and SR type index of dSR (1530, 1895), with R² values of 0.95 (p < 0.001) and 0.97 (p < 0.001), respectively, better than previously published indices. Even so, different best indices for different species were identified, most probably due to the differences in leaf anatomy and physiological processes during leaf dehydration. Although more plant species and field-measured datasets are still needed in future studies, the recommend indices based on derivative spectra provide a means to monitor drought-induced plant mortality in temperate climate regions.     

Allelopathic inhibition of phytoplankton by exudates from Stratiotes aloides

The allelopathic potential of exudates from the aquatic macrophyte Stratiotes aloides on the growth of phytoplankton was investigated. A selection of phytoplankton species, occurring in habitats similar to that of Stratiotes, was used: two cyanobacterial strains (toxic and non-toxic Microcystis aeruginosa), one green alga (Scenedesmus obliquus) and one eustigmatophyte (Nannochloropsis limnetica). The results indicate allelopathic effects of Stratiotes on phytoplankton in six of the eight cases, expressed in an extended duration of the initial biovolume doubling time. The overall inhibitory effect (8–51%) was strain-specific for the two cyanobacteria. We also studied the effect of irradiance on the allelopathic potential of exudates from Stratiotes. Irradiance influenced the response of Scenedesmus only. The inhibitory effect of Stratiotes exudates on the growth of this green alga was stronger at 35 μmol m⁻² s⁻¹ than at 105 μmol m⁻² s⁻¹. We conclude that Stratiotes has allelopathic effects on phytoplankton, and that irradiance can, but does not always determine the extent of the allelopathic inhibition. In our experiments, the sensitivity of cyanobacteria to Stratiotes exudates was not higher than for other phytoplankton strains, but within cyanobacteria, the toxic strain was more sensitive than the non-toxic one.     

Effects of nutrient-limiting supply ratios on toxin content of Karenia brevis grown in continuous culture

Toxins produced as secondary metabolites can play important roles in phytoplankton communities and contribute to the ecological success of harmful algal bloom (HAB) taxa. Toxin composition and content in phytoplankton are affected by a suite of environmental factors, including nutrient availability. Changes in nutrient availability can increase or decrease toxin content and alter toxin composition, depending on toxin stoichiometry and the mechanisms by which nutrient limitation affects toxin production. The studies that have assessed the effects of nutrient availability on brevetoxin content of the HAB species Karenia brevis have reported contradictory results, although there is growing support that nutrient limitation increases brevetoxin content. In this study, we assessed the effects of decreased nitrogen (N) and phosphorus (P) availability on brevetoxin content and composition of K. brevis grown in chemostats at steady state by altering the nutrient supply ratios of incoming media from the Redfield Ratio. Overall, brevetoxin content was greatest in cultures grown at the lowest rate, regardless of the nutrient supply ratio (i.e., under both Redfield and N-limiting supply ratios). Compared to cultures grown at 0.2 d⁻¹, cultures grown at 0.1 d⁻¹ exhibited 5-fold increases in intracellular toxin content. In contrast, at constant growth rates, N-limiting supply ratios decreased intracellular brevetoxin content by approximately one-third, although this result was significant only in cultures growing at the fastest rate of 0.23 d⁻¹. P-limiting supply ratios had no effect on brevetoxin content or composition. In addition, when cultures grown at rates of 0.2 d⁻¹ were supplied with balanced/Redfield N:P supply ratios, but different absolute nutrient concentrations, toxin content was greater under greater nutrient concentrations. These findings suggest that when growth rate is not nutrient limited, there is a positive relationship between nutrient availability and brevetoxin content. This work contributes to previous studies by demonstrating strong growth rates effects on brevetoxin content and that growth rate and nutrient availability can independently or together affect toxin content of K. brevis. Moreover, our work underscores the value of the chemostat as a tool to elucidate the mechanisms by which nutrient availability and growth rate affect toxin production and content of HAB species.     

Morphology and bloom dynamics of Cochlodinium geminatum (Schütt) Schütt in the Pearl River Estuary,South China Sea

An unarmored dinoflagellate bloom of Cochlodinium geminatum (Schütt) Schütt has been identified in the Pearl River Estuary, South China Sea during the severe dry season, from late October to early November, 2009, when temperature and salinity ranged between 20.0–27.2 °C and 10.6–33.4, respectively. Light and scanning electron microscopy were used to identify the characteristics of C. geminatum and provided the clear morphological structure for this species. The organism was primarily found in chains of two cells or single cell, and no longer chains were observed. Cells were irregularly spherical or slightly dorso-ventrally, with size ranged between 28 and 36 μm and longer than wide. A large nucleus in the center with numerous golden chloroplasts was present, and the cingulum made 1.5 turns around the cell. The concentration of C. geminatum ranged from 10² to greater than 10⁷ cells l⁻¹ during the bloom period. Nutrient concentration ranges during the bloom were 1.29–81.00 μM NO₃, 0.14–12.14 μM NO₂, 0.21–6.29 μM NH₄, 0.23–6.26 μM PO₄ and 3.29–171.43 μM SiO₃, respectively. Total biomass expressed in terms of chlorophyll a ranged from 2.44 to 135.45 μg l⁻¹, with an average 19.9 μg l⁻¹ in surface water throughout the PRE. Two main clusters corresponding to the water sectors were defined with multivariate analysis (cluster and nMDS). Based on the composition and abundance of phytoplankton, spatial variations were observed at a significant level (ANOSIM, R = 0.44, P < 0.01). Although the pairwise correlation analysis detected no significant effect of any single environmental variable on the abundance of C. geminatum, the multivariate analysis (BIO-ENV) between biotic and abiotic variables resulted in the best variables combination with all measured factors involved (temperature, salinity, turbidity, NO₃, NO₂, NH₄, PO₄ and SiO₃) which showed a combined effect during the bloom of C. geminatum in the Pearl River Estuary (ρ_w = 0.477).     

The distribution of chlorophyll a in the eastern equatorial Pacific Ocean in boreal autumn

The distribution and size fractions of chlorophyll a (Chl a) concentration in the eastern equatorial Pacific Ocean in boreal autumn were investigated during October and November, 2011. Environmental factors, including hydrology and nutrients, that might affect the distribution and size composition were analyzed. A total of 18 stations including 11 CTD stations and 7 navigation stations were selected which stretch from the northwest coast of South America to the area of the central Pacific Ocean south of the Hawaiian Islands (2.77°S–13.02°N, 84.11–154.02°W). The studied area can be divided into two transects: the 6°N transect (124–148°W) and the154°W transect (10–13°N). Results showed that the surface Chl a concentration was higher in the east near the northwest coast of South America (>0.200 mg/m³) and lower in the west (0.100–0.200 mg/m³), and it presented a highly significant negative correlation with sea surface temperature (p < 0.001). There were some differences between the sectional distribution of Chl a concentration between the 6°N and 154°W transects. The high values of Chl a concentration occurred near the surface along the 6°N transect (0–75 m), while they were relatively deeper along the 154°W transect (50–100 m). Iron might be the factor that limited the growth of phytoplankton in the eastern equatorial Pacific Ocean. Picophytoplankton (Pico) was the dominant taxa in the surveyed area, particularly in the waters along the two transects (>70% of total Chl a). The Pico to total Chl a ratio was higher in the upper layer (>70%) than in the deeper layer.     

Empirical models of toxigenic Pseudo-nitzschia blooms: Potential use as a remote detection tool in the Santa Barbara Channel

The Santa Barbara Channel, CA is a highly productive region where wind-driven upwelling and mesoscale eddies are important processes driving phytoplankton blooms. In recent years, the spring bloom has been dominated by the neurotoxin-producing diatom, Pseudo-nitzschia spp. In this paper, we relate a 1.5-year time series of Pseudo-nitzschia spp. abundance and domoic acid concentration to physical, chemical, and biological data to better understand the mechanisms controlling local Pseudo-nitzschia spp. bloom dynamics. The data were used to define the ranges of environmental conditions associated with Pseudo-nitzschia spp. bloom development in the Santa Barbara Channel. The time series captured three large toxic events (max. particulate domoic acid concentration, pDA ～6000 ng L^?1; max. cellular domoic acid concentrations, cDA ～88 pg cell^?1) in the springs of 2005–2006 and summer 2005 corresponding to bloom-level Pseudo-nitzschia spp. abundance (>5.0 × 10⁴ cells L^?1). In general, large increases in Pseudo-nitzschia spp. abundance were accompanied by increases in cDA levels, and cDA peaks preceded pDA peaks by at least one month in both the springs of 2005 and 2006. Statistical models incorporating satellite ocean color (MODIS-Aqua and SeaWiFS) and sea surface temperature (AVHRR) data were created to determine the probability that a remotely sensed phytoplankton bloom contains a significant population of toxic Pseudo-nitzschia spp. Models correctly estimate 98% of toxic bloom situations, with a 7–29% rate of false positive identification. Conditions most associated with high cDA levels are low sea surface temperature, high salinity, increased absorption by cDOM (412 nm), increased reflectance at 510/555 nm, and decreased particulate absorption at 510 nm. Future efforts to merge satellite and regionally downscaled forecasting products with these habitat models will help assess bloom forecasting capabilities in the central CA region and any potential connections to large-scale climate modes.     

Photoadaptation in marine phytoplankton : changes in spectral absorption and excitation of chlorophyll a fluorescence

The optical properties of marine phytoplankton were examined by measuring the absorption spectra and fluorescence excitation spectra of chlorophyll a for natural marine particles collected on glass fiber filters. Samples were collected at different depths from stations in temperate waters of the Southern California Bight and in polar waters of the Scotia and Ross Seas. At all stations, phytoplankton fluorescence excitation and absorption spectra changed systematically with depth and vertical stability of the water columns. In samples from deeper waters, both absorption and chlorophyll a fluorescence excitation spectra showed enhancement in the blue-to-green portion of the spectrum (470-560 nm) relative to that at 440 nm. Since similar changes in absorption and excitation were induced by incubating sea water samples at different light intensities, the changes in optical properties can be attributed to photoadaptation of the phytoplankton. The data indicate that in the natural populations studied, shade adaptation caused increases in the concentration of photosynthetic accessory pigments relative to chlorophyll a. These changes in cellular pigment composition were detectable within less than 1 day. Comparisons of absorption spectra with fluorescence excitation spectra indicate an apparent increase in the efficiency of sensitization of chlorophyll a fluorescence in the blue and green spectral regions for low light populations.     

Structured near-infrared Magnetic Circular Dichroism spectra of the Mn4CaO5 cluster of PSII in T. vulcanus are dominated by Mn(IV) d-d ‘spin-flip’ transitions

Photosystem II passes through four metastable S-states in catalysing light-driven water oxidation. Variable temperature variable field (VTVH) Magnetic Circular Dichroism (MCD) spectra in PSII of Thermosynochococcus (T.) vulcanus for each S-state are reported. These spectra, along with assignments, provide a new window into the electronic and magnetic structure of Mn₄CaO₅. VTVH MCD spectra taken in the S₂ state provide a clear g = 2, S = 1/2 paramagnetic characteristic, which is entirely consistent with that known by EPR. The three features, seen as positive (+) at 749 nm, negative (?) at 773 nm and (+) at 808 nm are assigned as ⁴A → ²E spin-flips within the d³ configuration of the Mn(IV) centres present. This assignment is supported by comparison(s) to spin-flips seen in a range of Mn(IV) materials. S₃ exhibits a more intense (?) MCD peak at 764 nm and has a stronger MCD saturation characteristic. This S₃ MCD saturation behaviour can be accurately modelled using parameters taken directly from analyses of EPR spectra. We see no evidence for Mn(III) d-d absorption in the near-IR of any S-state. We suggest that Mn(IV)-based absorption may be responsible for the well-known near-IR induced changes induced in S₂ EPR spectra of T. vulcanus and not Mn(III)-based, as has been commonly assumed. Through an analysis of the nephelauxetic effect, the excitation energy of S-state dependent spin-flips seen may help identify coordination characteristics and changes at each Mn(IV). A prospectus as to what more detailed S-state dependent MCD studies promise to achieve is outlined.     

Genome size variation and polyploidy in the resurrection plant genus Ramonda: Cytogeography of living fossils

The genus Ramonda includes three preglacial paleoendemic species surviving as the rare resurrection angiosperms of the Northern hemisphere in refugia habitats in the Balkan (Ramonda nathaliae and Ramonda serbica) and Iberian Peninsulas (Ramonda myconi). This study focuses on: assessing genome size and base composition, determining chromosome number and ploidy level in several populations, evaluating inter- and intra-specific variations in DNA content and chromosome number as well as looking for the possible hybridization in the sympatric zones of Balkan species. R. nathaliae and R. myconi are diploid species (2n = 2x = 48) while R. serbica is hexaploid (2n = 6x = 144). The mean 2C DNA values ranged from 2.30 pg for R. nathaliae to 2.59 pg for R. myconi compared to 7.91 pg for R. serbica. The base composition for R. nathaliae was 42.1% GC, for R. myconi 39.9% and for R. serbica 41.2%. In one population of R. serbica the DNA content ranged from 2C = 7.65 to 11.82 pg, revealing different ploidy levels among its individuals. In sympatric populations genome size was intermediary (～5 pg) between the diploid and hexaploid classes which indicates the hybridization ability between R. serbica and R. nathaliae. It appears that polyploidization is the major evolutionary mechanism in the genus Ramonda.     

The rate-limiting step in O2 reduction by cytochrome ba3 from Thermus thermophilus

Cytochrome ba₃ (ba₃) of Thermus thermophilus (T. thermophilus) is a member of the heme–copper oxidase family, which has a binuclear catalytic center comprised of a heme (heme a₃) and a copper (Cu_B). The heme–copper oxidases generally catalyze the four electron reduction of molecular oxygen in a sequence involving several intermediates. We have investigated the reaction of the fully reduced ba₃ with O₂ using stopped-flow techniques. Transient visible absorption spectra indicated that a fraction of the enzyme decayed to the oxidized state within the dead time (~ 1 ms) of the stopped-flow instrument, while the remaining amount was in a reduced state that decayed slowly (k = 400 s^? 1) to the oxidized state without accumulation of detectable intermediates. Furthermore, no accumulation of intermediate species at 1 ms was detected in time resolved resonance Raman measurements of the reaction. These findings suggest that O₂ binds rapidly to heme a₃ in one fraction of the enzyme and progresses to the oxidized state. In the other fraction of the enzyme, O₂ binds transiently to a trap, likely Cu_B, prior to its migration to heme a₃ for the oxidative reaction, highlighting the critical role of Cu_B in regulating the oxygen reaction kinetics in the oxidase superfamily. This article is part of a Special Issue entitled: Respiratory Oxidases.     

Habitat associations of an insular Wallacean avifauna: A multi-scale approach for biodiversity proxies

The endemic avifauna of Wallacea is of high conservation significance, but remains poorly studied. Identifying priority conservation areas requires a greater understanding of the habitat associations of these bird communities, and of how spatial scale of analysis can influence the interpretation of these associations. This study aims to determine which proxy habitat measures, at which spatial scales of analysis, can provide useful inferential data on the composition of Wallacean forest avifauna. Research was conducted within the Lambusango forest reserve, South-East Sulawesi, using point count surveys to sample avifauna. Habitat properties were characterised in three ways: broad classification of forest type, canopy remotely-sensed response derived from satellite imagery, and in situ measures of vegetation composition and structure. Furthermore, we examined avifauna–habitat relationships at three spatial scales: area (c.400 ha per sample site), transect (c.10 ha) and point (c.0.2 ha). Results demonstrate that broad forest type classifications at an area scale can help to determine conservation value, indicating that primary and old secondary forests are important for supporting many species with lower ecological tolerances, such as large-bodied frugivores. At the transect-scale, significant congruence occurs between bird community composition and several habitat variables derived from vegetation sampling and satellite imagery, particularly tree size, undergrowth density, and Normalised Difference Vegetation Index (NDVI) values; this highlights the importance small scale habitat associations can have on determining α-diversity. Analysis at the point-scale was ineffective in providing proxy indications for avifauna. These findings should be considered when determining future priority conservation areas for Wallacean avifauna.     

The applicability of empirical vegetation indices for determining leaf chlorophyll content over different leaf and canopy structures

Retrieving leaf chlorophyll content at a range of spatio-temporal scales is central to monitoring vegetation productivity, identifying physiological stress and managing biological resources. However, estimating leaf chlorophyll over broad spatial extents using ground-based traditional methods is time and resource heavy. Satellite-derived spectral vegetation indices (VIs) are commonly used to estimate leaf chlorophyll content, however they are often developed and tested on broadleaf species. Relatively little research has assessed VIs for different leaf structures, particularly needle leaves which represent a large component of boreal forest and significant global ecosystems. This study tested the performance of 47 published VIs for estimating foliar chlorophyll content from different leaf and canopy structures (broadleaf and needle). Coniferous and deciduous sites were selected in Ontario, Canada, representing different dominant vegetation species (Picea mariana and Acer saccharum) and a variety of canopy structures. Leaf reflectance data was collected using an ASD Fieldspec Pro spectroradiometer (400–2500 nm) for over 300 leaf samples. Canopy reflectance data was acquired from the medium resolution imaging spectrometer (MERIS). At the canopy level, with both leaf types combined, the DD-index showed the strongest relationship with leaf chlorophyll (R² = 0.78; RMSE = 3.56 μg/cm²), despite differences in leaf structure. For needleleaf trees alone the relationship with the top VI was weaker (D_[red], R² = 0.71; RMSE = 2.32 μg/cm²). A sensitivity study using simulated VIs from physically-modelled leaf (PROSPECT) and canopy (4-Scale) reflectance was performed in order to further investigate these results and assess the impacts of different background types and leaf area index on the VIs’ performance. At the leaf level, the MNDVI8 index showed a strong linearity to changing chlorophyll and negligible difference to leaf structure/type. At canopy level, the best performing VIs were relatively consistent where LAI ≥ 4, but responded strongly to differences in background at low canopy coverage (LAI = 2). This research provides comprehensive assessments for the use of spectral indices in retrieval of spatially-continuous leaf chlorophyll content at the leaf (MTCI: R² = 0.72; p < 0.001) and canopy (DD: R² = 0.78; p < 0.001) level for resource management over different spatial and temporal scales.     

Is inorganic nutrient enrichment a driving force for the formation of red tides?: A case study of the dinoflagellate Scrippsiella trochoidea in an embayment

Red tides (high biomass phytoplankton blooms) have frequently occurred in Hong Kong waters, but most red tides occurred in waters which are not very eutrophic. For example, Port Shelter, a semi-enclosed bay in the northeast of Hong Kong, is one of hot spots for red tides. Concentrations of ambient inorganic nutrients (e.g. N, P), are not high enough to form the high biomass of chlorophyll a (chl a) in a red tide when chl a is converted to its particulate organic nutrient (N) (which should equal the inorganic nutrient, N). When a red tide of the dinoflagellate Scrippsiella trochoidea occurred in the bay, we found that the red tide patch along the shore had a high cell density of 15,000 cells ml⁻¹, and high chl a (56 μg l⁻¹), and pH reached 8.6 at the surface (8.2 at the bottom), indicating active photosynthesis in situ. Ambient inorganic nutrients (NO₃, PO₄, SiO₄, and NH₄) were all low in the waters and deep waters surrounding the red tide patch, suggesting that the nutrients were not high enough to support the high chl a >50 μg l⁻¹ in the red tide. Nutrient addition experiments showed that the addition of all of the inorganic nutrients to a non-red-tide water sample containing low concentrations of Scrippsiella trochoidea did not produce cell density of Scrippsiella trochoidea as high as in the red tide patch, suggesting that nutrients were not an initializing factor for this red tide. During the incubation of the red tide water sample without any nutrient addition, the phytoplankton biomass decreased gradually over 9 days. However, with a N addition, the phytoplankton biomass increased steadily until day 7, which suggested that nitrogen addition was able to sustain the high biomass of the red tide for a week with and without nutrients. In contrast, the red tide in the bay disappeared on the sampling day when the wind direction changed. These results indicated that initiation, maintenance and disappearance of the dinoflagellate Scrippsiella trochoidea red tide in the bay were not directly driven by changes in nutrients. Therefore, how nutrients are linked to the formation of red tides in coastal waters need to be further examined, particularly in relation to dissolved organic nutrients.