Multispectral image analysis for algal biomass quantification

This article reports a novel multispectral image processing technique for rapid, noninvasive quantification of biomass concentration in attached and suspended algae cultures. Monitoring the biomass concentration is critical for efficient production of biofuel feedstocks, food supplements, and bioactive chemicals. Particularly, noninvasive and rapid detection techniques can significantly aid in providing delay‐free process control feedback in large‐scale cultivation platforms. In this technique, three‐band spectral images of Anabaena variabilis cultures were acquired and separated into their red, green, and blue components. A correlation between the magnitude of the green component and the areal biomass concentration was generated. The correlation predicted the biomass concentrations of independently prepared attached and suspended cultures with errors of 7 and 15%, respectively, and the effect of varying lighting conditions and background color were investigated. This method can provide necessary feedback for dilution and harvesting strategies to maximize photosynthetic conversion efficiency in large‐scale operation. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:808–816, 2013     

Differentiating aquatic plant communities in a eutrophic river using hyperspectral and multispectral remote sensing

1. This study evaluates the efficacy of remote sensing technology to monitor species composition, areal extent and density of aquatic plants (macrophytes and filamentous algae) in impoundments where their presence may violate water‐quality standards. 2. Multispectral satellite (IKONOS) images and more than 500 in situ hyperspectral samples were acquired to map aquatic plant distributions. By analyzing field measurements, we created a library of hyperspectral signatures for a variety of aquatic plant species, associations and densities. We also used three vegetation indices. Normalized Difference Vegetation Index (NDVI), near‐infrared (NIR)‐Green Angle Index (NGAI) and normalized water absorption depth (D_H)_, at wavelengths 554, 680, 820 and 977 nm to differentiate among aquatic plant species composition, areal density and thickness in cases where hyperspectral analysis yielded potentially ambiguous interpretations. 3. We compared the NDVI derived from IKONOS imagery with the in situ, hyperspectral‐derived NDVI. The IKONOS‐based images were also compared to data obtained through routine visual observations. Our results confirmed that aquatic species composition alters spectral signatures and affects the accuracy of remote sensing of aquatic plant density. The results also demonstrated that the NGAI has apparent advantages in estimating density over the NDVI and the D_H. 4. In the feature space of the three indices, 3D scatter plot analysis revealed that hyperspectral data can differentiate several aquatic plant associations. High‐resolution multispectral imagery provided useful information to distinguish among biophysical aquatic plant characteristics. Classification analysis indicated that using satellite imagery to assess Lemna coverage yielded an overall agreement of 79% with visual observations and >90% agreement for the densest aquatic plant coverages. 5. Interpretation of biophysical parameters derived from high‐resolution satellite or airborne imagery should prove to be a valuable approach for assessing the effectiveness of management practices for controlling aquatic plant growth in inland waters, as well as for routine monitoring of aquatic plants in lakes and suitable lentic environments.     

基于多光谱影像的森林树种识别及其空间尺度响应

当前,不同空间分辨率卫星影像对森林类型识别结果中普遍存在的尺度效应,而且纹理参量对不同尺度下树种识别精度的影响仍缺乏广泛认知.本研究以中国东北旺业甸林场为研究区,采用观测时相同步、地理坐标匹配的GF-1 PMS、GF-2 PMS、GF-1 WFV,以及Landsat-8 OLI卫星传感器数据组成空间尺度观测序列(1、2、4、8、16、30 m),并结合支持向量机(SVM)模型,探讨了区域内5种优势树种遥感识别结果的尺度变化规律及其纹理特征参数的影响,同时检验了基于尺度上推转换影像的树种识别结果差异.结果表明: 影像空间分辨率对区域树种识别结果具有显著影响,其中,研究区森林树种识别的最佳影像分辨率为4 m,当分辨率降低至30 m时,树种识别结果最差.在1～8 m影像分辨率范围内,增加纹理信息能够显著提高不同优势树种的识别精度,使总分类精度提升了2.0%～3.6%,但纹理信息对16～30 m影像的识别结果没有显著影响.与真实尺度卫星影像相比,基于升尺度转换影像的树种识别结果及其尺度响应特征存在显著差异,表明在面向多个空间尺度的遥感观测和应用研究中,需要采用真实分辨率影像以确保结果的准确性.     

Rapid expansion of the invasive-like red macroalga,Chondria tumulosa (Rhodophyta), on the coral reefs of the Papahānaumokuākea Marine National Monument

A cryptogenic, invasive-like red macroalga, Chondria tumulosa, was first observed in 2016 forming thick mats on the forereef of Manawai Atoll within Papahānaumokuākea Marine National Monument. Subsequent expeditions revealed an increased abundance of this alga. In 2021, unattached C. tumulosa was observed forming a network of dark, meandering accumulations throughout the atoll's inner lagoon. High-resolution satellite imagery revealed that these accumulations became visible in 2015 (length: ~0.74 km; area: ~0.88 km²) and increased 56-fold in length and 115-fold in area by 2021 (length: 41.32 km; area: 101.34 km²). An exponential expansion rate of ~16.02 km · y⁻¹ (length), ~44.75 km² · y⁻¹ (area). This study presents the comprehensive temporal and spatial expansion of C. tumulosa accumulations for Manawai Atoll since its discovery, providing ecologist and resource managers with a proxy to gauge the overall abundance trend of this invasive-like alga.     

Identifying wetland meadows in Grand Teton National Park using remote sensing and average wetland values

Six spectrally and ecologically distinct montane meadow community types were identified and mapped within Grand Teton National Park by analysis of Indian IRS-1B LISS-II imagery. A distinct to-xeric-hydric gradient among the meadow types was predicted by analysis of the satellite data. Thirty sites (five replicates for each of six meadow typ were selected for intensive field sampling. At each of the 30 sites, meadow vegetation was sampled in 20 m by 20 m square plots for canopy cover of all species. Using wetland indexes (on a scale of 1–5, where obligate wetland species = 1, facultative land = 2, facultative = 3, facultative upland = 4 and upland species = 5), average wetland values were calculated and ranged from 1.88 for A-type meadows and 2.86 for B meadows to 4.40, 4.49, 4.74, and 4.43 for C, D, E and F meadows, respectively. Because average wetland values of A and B meadows were < 3.00, they were determined to be indicative of wetlands. Eight out of ten obligate wetland plants had their greatest cover on A meadows (the wettest of the gradient) and had significant cover differe among meadow types using the non-parametric Kruskal-Wallis test. Average wetland values and plant species cover were used, in conjunction with remotely sensed data, to identify as wetlands 1,258 hectares of A meadows and 1,711 hectares of B meadows in Grand Teton National Park.     

A bird's-eye view: Evaluating drone imagery for the detection and monitoring of endangered and invasive day geckos

Species monitoring can be strongly limited by terrain accessibility, impeding our understanding of species ecology and thus challenging their conservation. This is particularly true for species living in the canopy, on cliffs or in dense vegetation. Remote sensing imagery may fill this gap by offering a cost-effective monitoring approach allowing to improve species detection in inaccessible areas. We investigated the applicability of drone-based monitoring for a Critically Endangered insular gecko (Phelsuma inexpectata) and two invasive alien gecko species representing a risk for the former (P. grandis and P. laticauda). We determined the approach distance before inducing behavioural response caused by the drone's presence. All three study species showed no escape behaviour to the drone's presence until very close distances (mean distance for P. inexpectata: 33.8 cm; P. grandis: 21.9 cm; P. laticauda: 26.4 cm). We then performed horizontal and vertical approaches with the drone, taking photos every meter starting at 10 m away from the canopy edge to determine an optimal distance for detection while ensuring species-level identification. We examined a total of 328 photos. We found a bimodality in the number of detected geckos for two species, with different individuals recorded between short and intermediate distances. Therefore, we recommend taking photos at two distances of 2–2.5 and 5 m away from the canopy, ideally facing away from the sun and in low wind conditions. We encourage the application of our methodology for Phelsuma spp., but also for other species of similar size and ecology to improve detection in inaccessible areas.     

Low-light image analysis of transgenic organisms using bacterial luciferase as a marker

Methods for measurement of a novel light-emitting reporter gene system in bacteria, yeast, plant cells, plant tissues and intact plant organs are described. The principle underlying the assay procedures is the bacterial luciferase catalysed oxidation of reduced flavin mononucleotide (FMNH₂) in the presence of the ten carbon aldehyde decanal, to yield FMN, decanoic acid, water and a photon of light at 490 nm which can be captured by X-ray film, a photomultiplier tube or, for in vivo measurements, an image-intensifier coupled to a video camera. This light measuring assay system is sensitive, easy to use, inexpensive, does not require radioactivity, and has been used successfully for rapid detection of bacterial transformants, the quantitative measurement of transient and stable gene expression in bacteria and yeast, and in vivo measurement of temporal and spatial gene expression throughout plant and animal development.     

Three-Dimensional (3D) cell culture monitoring: Opportunities and challenges for impedance spectroscopy

Three-dimensional (3D) cell culture has developed rapidly over the past 5–10 years with the goal of better replicating human physiology and tissue complexity in the laboratory. Quantifying cellular responses is fundamental in understanding how cells and tissues respond during their growth cycle and in response to external stimuli. There is a need to develop and validate tools that can give insight into cell number, viability, and distribution in real-time, nondestructively and without the use of stains or other labelling processes. Impedance spectroscopy can address all of these challenges and is currently used both commercially and in academic laboratories to measure cellular processes in 2D cell culture systems. However, its use in 3D cultures is not straight forward due to the complexity of the electrical circuit model of 3D tissues. In addition, there are challenges in the design and integration of electrodes within 3D cell culture systems. Researchers have used a range of strategies to implement impedance spectroscopy in 3D systems. This review examines electrode design, integration, and outcomes of a range of impedance spectroscopy studies and multiparametric systems relevant to 3D cell cultures. While these systems provide whole culture data, impedance tomography approaches have shown how this technique can be used to achieve spatial resolution. This review demonstrates how impedance spectroscopy and tomography can be used to provide real-time sensing in 3D cell cultures, but challenges remain in integrating electrodes without affecting cell culture functionality. If these challenges can be addressed and more realistic electrical models for 3D tissues developed, the implementation of impedance-based systems will be able to provide real-time, quantitative tracking of 3D cell culture systems.     

Towards a whole-system framework for wildfire monitoring using Earth observations

Fire seasons have become increasingly variable and extreme due to changing climatological, ecological, and social conditions. Earth observation data are critical for monitoring fires and their impacts. Herein, we present a whole-system framework for identifying and synthesizing fire monitoring objectives and data needs throughout the life cycle of a fire event. The four stages of fire monitoring using Earth observation data include the following: (1) pre-fire vegetation inventories, (2) active-fire monitoring, (3) post-fire assessment, and (4) multi-scale synthesis. We identify the challenges and opportunities associated with current approaches to fire monitoring, highlighting four case studies from North American boreal, montane, and grassland ecosystems. While the case studies are localized to these ecosystems and regional contexts, they provide insights for others experiencing similar monitoring challenges worldwide. The field of remote sensing is experiencing a rapid proliferation of new data sources, providing observations that can inform all aspects of our fire monitoring framework; however, significant challenges for meeting fire monitoring objectives remain. We identify future opportunities for data sharing and rapid co-development of information products using cloud computing that benefits from open-access Earth observation and other geospatial data layers.     

Computer-based image analysis for the automated counting and morphological description of microalgae in culture

A largely unexplored area is the application of digital image processing to counting and sizing of microalgal cells from culture. Commercial systems are available, but have not been tested, nor necessarily optimized for high speed counting and sizing of phytoplankton. The present work describes the design, construction, specifications and comparative performance of an inexpensive system optimized for counting and sizing microalgal cells. This system has been tested with cells of the picoplankton to nanoplankton size ranges (1–20 μm). The hardware was a widely available standard microcomputer, an inexpensive video camera and monitor, and a video digitization board (frame grabber). A modifiable menu-driven program (PHYCOUNT) was written and provisions made to make this program available to other workers. The program is constructed such that it can be adapted to a variety of hardware setups Video digitization boards). Comparison of growth curves for microagae revealed there were no significant differences in division rate and cell yield as assessed by the image analysis method compared to manual counts with a hemacytometer. Several hundred cells were counted routinely within 10–15 s, far exceeding the counting rate achieved by hand tally. A variable transect feature allowed sampling every nth pixel and provided a substantial increase in execution speed. More than 1000 counts can be done per day. A protocol for the use of 96-well plates of polyvinyl chloride as counting chambers contributed to the processing of large numbers of samples rapidly. Other routines developed provided subtended area, defined the coordinates of cell perimeter, and derived cell length and width. The calculation of the latter two parameters was usually done off-line as data output is in standard numerical form accessible by other programs. Experience with daily use of the PHYCOUNT program and imaging hardware reveal that the system is reliable for cell counting and sizing. The presence of bacteria in the algal cultures does not affect cell counting or sizing.     

New bacterial culture medium for production of mosquito pathogenic bacilli using agro-poultry industrial wastes

A cost-effective technology has been developed to utilise bioorganic wastes as culture media, to produce mosquitocidal biopesticides, Bacillus sphaericus and B. thuringiensis serovar israelensis. The mosquitocidal spore/crystal toxins produced from the experimental medium (chicken feather waste, CFW+paddy husk waste, PHW) was higher than that of the conventional medium (Nutrient Yeast Extract Salt Medium, NYSM). The bacterial toxins produced from different media (NYSM, CFW, PHW, CFW+PHW) were bioassayed against mosquito vectors and the toxic effect was found to be significant. The use of chicken feathers and paddy husk wastes as bacterial culture media is cost-effective and economical for the production of these mosquito pathogenic bacilli.     

Secretome analysis using a hollow fiber culture system for cancer biomarker discovery

Secreted proteins, collectively referred to as the secretome, were suggested as valuable biomarkers in disease diagnosis and prognosis. However, some secreted proteins from cell cultures are difficult to detect because of their intrinsically low abundance; they are frequently masked by the released proteins from lysed cells and the substantial amounts of serum proteins used in culture medium. The hollow fiber culture (HFC) system is a commercially available system composed of small fibers sealed in a cartridge shell; cells grow on the outside of the fiber. Recently, because this system can help cells grow at a high density, it has been developed and applied in a novel analytical platform for cell secretome collection in cancer biomarker discovery. This article focuses on the advantages of the HFC system, including the effectiveness of the system for collection of secretomes, and reviews the process of cell secretome collection by the HFC system and proteomic approaches to discover cancer biomarkers. The HFC system not only provides a high-density three-dimensional (3D) cell culture system to mimic tumor growth conditions in vivo but can also accommodate numerous cells in a small volume, allowing secreted proteins to be accumulated and concentrated. In addition, cell lysis rates can be greatly reduced, decreasing the amount of contamination by abundant cytosolic proteins from lysed cells. Therefore, the HFC system is useful for preparing a wide range of proteins from cell secretomes and provides an effective method for collecting higher amounts of secreted proteins from cancer cells. This article is part of a Special Issue entitled: An Updated Secretome.     

三种回归分析方法在Hyperion影像LAI反演中的比较

借助GPS进行地面精确定位,利用LAI-2000冠层分析仅在攸县黄丰桥林场开展130个样地(60m×60m)的叶面积指数(Leaf Area Index,LAI)测量.采用FLAASH模块对Hyperion数据进行大气校正并与地面同步冠层观测数据进行拟合,通过研究地面实测LAI与Hyperion影像波段及其衍生的系列植被指数(NDVI、RVI等)的相关性,筛选出估算叶面积指数的植被指数因子.应用曲线估计、逐步回归及偏最小二乘三种回归分析技术分别建立叶面积指数的最优估算模型.结果表明:参与建模的因子中,比值植被指数(RVI)与LAI的相关性最大,敏感性最高,其次是SARVI0.1,NDVI705,NDVI,SARVI0.1,SARVI0.25;曲线估计、逐步回归分析和偏最小二乘回归三种分析方法所建的6个回归模型中,偏最小二乘回归的拟合效果最好,预测值与实测值的决定系数R2为0.84、曲线估计的拟合效果最低,预测值与实测值的决定系数R2为0.64;建模精度分析表明,选用5-6个自变量因子进行LAI建模是可靠的,以6个植被因子建立的偏最小二乘回归模型预测精度最高.     

New method for forming large embryoid bodies using the wall of the culture dish along with an analysis of their structural characteristics

Early embryonic stem (EES) cells, which were established from 2 cell stage embryos obtained from ddY mice, had similar characteristics as embryonic stem (ES) cells. These cells were maintained in an undifferentiated stage in growth media supplemented with leukemia inhibitory factor (LIF) and were capable of differentiating into triploblastic tissues under various growth factors. It has been known that normal sized embryoid bodies (EBs) are formed by removing LIF. In this study, large EBs gradually formed along the side wall of a culture dish, particularly at the boundary between the air and the growth medium when cells were cultured for a considerable period of time and without subculturing. We call this method the "wall adhesion culture" procedure. The method itself is simple and do not need any instruments except plastic dishes because only the side walls of the dishes were utilized. The mean thickness of the large EBs was about 1.5 mm 3 months after establishing the static culture. Their surface was covered with a monolayer of cells and they contained an eosinophilic cell matrix. By electron microscopy, some characteristic structures was observed, such as intracisternal A particles which were present inside the swelling of the rough endoplasmic reticulum. Since many tissues derived from ES cells are obtained through EBs, it is expected that efficient acquisition of sufficient quantities of these structures using the wall adhesion culture procedure will be a shortcut for using ES cells in regenerative medicine.     

Temporal transferability of wildlife habitat models: implications for habitat monitoring

Aim Temporal transferability is an important issue when habitat models are used beyond the time frame corresponding to model development, but has not received enough attention, particularly in the context of habitat monitoring. While the combination of remote sensing technology and habitat modelling provides a useful tool for habitat monitoring, the effect of incorporating remotely sensed data on model transferability is unclear. Therefore, our objectives were to assess how different satellite‐derived variables affect temporal transferability of habitat models and their usefulness for habitat monitoring. Location Wolong Nature Reserve, Sichuan Province, China. Methods We modelled giant panda habitat with the maximum entropy algorithm using panda presence data collected in two time periods and four different sets of predictor variables representing land surface phenology. Each predictor variable set contained either a time series of smoothed wide dynamic range vegetation index (WDRVI) or 11 phenology metrics, both derived from single‐year or multi‐year (i.e. 3‐year) remotely sensed imagery acquired by the Moderate Resolution Imaging Spectroradiometer (MODIS). We evaluated the ability of models obtained with these four variable sets to predict giant panda habitat within and across time periods by using threshold‐independent and threshold‐dependent evaluation methods and five indices of temporal transferability. Results Our results showed that models developed with the four variable sets were all useful for characterizing and monitoring giant panda habitat. However, the models developed using multi‐year data exhibited significantly higher temporal transferability than those developed using single‐year data. In addition, models developed with phenology metrics, especially when using multi‐year data, exhibited significantly higher temporal transferability than those developed with the time series. Main conclusions The integration of land surface phenology, captured by high temporal resolution remotely sensed imagery, with habitat modelling constitutes a suitable tool for characterizing wildlife habitat and monitoring its temporal dynamics. Using multi‐year phenology metrics reduces model complexity, multicollinearity among predictor variables and variability caused by inter‐annual climatic fluctuations, thereby increasing the temporal transferability of models. This study provides useful guidance for habitat monitoring through the integration of remote sensing technology and habitat modelling, which may be useful for the conservation of the giant panda and many other species.     

A high‐resolution approach for the spatiotemporal analysis of forest canopy space using terrestrial laser scanning data

Forest canopies and tree crown structures are of high ecological importance. Measuring canopies and crowns by direct inventory methods is time‐consuming and of limited accuracy. High‐resolution inventory tools, in particular terrestrial laser scanning (TLS), is able to overcome these limitations and obtain three‐dimensional (3D) structural information about the canopy with a very high level of detail. The main objective of this study was to introduce a novel method to analyze spatiotemporal dynamics in canopy occupancy at the individual tree and local neighborhood level using high‐resolution 3D TLS data. For the analyses, a voxel grid approach was applied. The tree crowns were modeled through the combination of two approaches: the encasement of all crown points with a 3D α‐shape, which was then converted into a voxel grid, and the direct voxelization of the crown points. We show that canopy occupancy at individual tree level can be quantified as the crown volume occupied only by the respective tree or shared with neighboring trees. At the local neighborhood level, our method enables the precise determination of the extent of canopy space filling, the identification of tree–tree interactions, and the analysis of complementary space use. Using multitemporal TLS data recordings, this method allows the precise detection and quantification of changes in canopy occupancy through time. The method is applicable to a wide range of investigations in forest ecology research, including the study of tree diversity effects on forest productivity or growing space analyses for optimal tree growth. Due to the high accuracy of this novel method, it facilitates the precise analyses even of highly plastic individual tree crowns and, thus, the realistic representation of forest canopies. Moreover, our voxel grid framework is flexible enough to allow for the inclusion of further biotic and abiotic variables relevant to complex analyses of forest canopy dynamics.     

Vegetation–Climate Interactions among Native and Invasive Species in Hawaiian Rainforest

We compiled a time series of Earth Observing-1 Hyperion satellite observations with field measurements to compare the structural, biochemical, and physiological characteristics of an invasive nitrogen-fixing tree Myrica faya and native Metrosideros polymorpha in montane rainforests in Hawai’i. Satellite-based canopy water measurements closely tracked variations in leaf area index, and the remotely sensed photochemical and carotenoid reflectance indices (PRI, CRI) indicated variations in upper-canopy leaf chlorophyll and carotenoid content during a climatological transition. The PRI and CRI were related to differences in light-use efficiency of each species, as indicated by field measurements of leaf electron transport rate. The suite of hyperspectral metrics indicated maximum differences in the structure, biochemistry, and physiology of Myrica and Metrosideros when canopy vapor pressure deficit was high during hotter and drier periods. These satellite data, combined with the Carnegue-Ames-Stanford Approach (CASA) carbon cycle model, suggested that Myrica growth rates were 16–44% higher than Metrosideros, with relative differences between species closely linked to climate conditions. The satellite hyperspectral data identified the basic biological mechanisms favoring the spread of an introduced tree, and provided a more detailed understanding of how vegetation–climate interactions affect the time course of plant growth with respect to the invasion process.