Search results for: Dinoflagellate
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 12

Search results for: Dinoflagellate

12 Alexandrium pacificum Cysts Distribution in One North African Lagoon Ecosystem

Authors: M. Fertouna Bellakhal, M. Bellakhal, A. Dhib, A. Fathalli, S. Turki, L. Aleya

Abstract:

Study of dinoflagellate cysts is a precious tool to get information about environment and water quality in many aquatic ecosystems. The distribution of Alexandrium pacificum cysts, in Bizerta lagoon located in North of Tunisia, was made based on sediment samples analysis from 123 equidistant stations delimiting 125 km² surfaces. Sediment characteristics such as percentage of water, organic matter, and particle size were analyzed to determine the factors that influence the distribution of this dinoflagellate. In addition, morphological examination and ribotyping of vegetative forms from microalgal cultures made from cyst germination confirmed the identity of the species attributed to A. pacificum. A correlation between the abundance of A. pacificum cysts and the percentage of water and sediment organic matter was recorded. In addition, the sedimentary fraction < 63μm was found to be potentially favorable for the installation and initiation of the Alexandrium pacificum efflorescence at the Bizerte lagoon. The mapping of cysts in this aquatic ecosystem has also allowed us to define distinct areas with specific abundance with closed relationship with shellfish aquaculture stations located within the lagoon.

Keywords: Alexandrium pacificum, cysts, Dinoflagellate, microalgal culture

Procedia PDF Downloads 120
11 Environmental Impact of Cysts of Some Dinoflagellates Species in the Bizerta Lagoon

Authors: M. Bellakhal, M. Bellakhal, L. Aleya

Abstract:

The specific composition and abundance of dinoflagellate resistance cysts in relation to environmental factors were studied from the superficial sediment at 123 stations in the Bizerte lagoon. 48 morphotypes of dinoflagellate cysts were identified, mainly dominated by Brigantidinium simplex, Votadinum spinosum, Alexandrium pacificum, Alexandrium pseudogonyaulax, and Lingulodinum machaerophorum. The density of cysts ranged from 1276 to 20126 cysts g⁻¹ dry sediment. Significant differences in the distribution pattern of the cysts were recorded, which allowed us to distinguish two areas; thus the inner areas of the lagoon have an abundance of cysts greater than the areas with marine influence. Ballast water discharges and shellfish culture may be incriminated as potential sources of introduction of species, particularly potentially toxic ones such as A. pacificum and Polysphaeridium zoharyi, without neglecting the role of currents in cyst distribution. Cyst mapping can be used as an indicator of potential foci of future toxic species blooms in this ecosystem.

Keywords: Bizerta Lagoon, cysts, dinoflagellates, mapping

Procedia PDF Downloads 96
10 Last ca 2500 Yr History of the Harmful Algal Blooms in South China Reconstructed on Organic-Walled Dinoflagellate Cysts

Authors: Anastasia Poliakova

Abstract:

Harmful algal bloom (HAB) is a known negative phenomenon that is caused both by natural factors and anthropogenic influence. HABs can result in a series of deleterious effects, such as beach fouling, paralytic shellfish poisoning, mass mortality of marine species, and a threat to human health, especially if toxins pollute drinking water or occur nearby public resorts. In South China, the problem of HABs has an ultimately important meaning. For this study, we used a 1.5 m sediment core LAX-2018-2 collected in 2018 from the Zhanjiang Mangrove National Nature Reserve (109°03´E, 20°30´N), Guangdong Province, South China. High-resolution coastal environment reconstruction with a specific focus on the HABs history during the last ca 2500 yrs was attempted. Age control was performed with five radiocarbon dates obtained from benthic foraminifera. A total number of 71 dinoflagellate cyst types was recorded. The most common types found consistently throughout the sediment sequence were autotrophic Spiniferites spp., Spiniferites hyperacanthus and S. mirabilis, S. ramosus, Operculodinium centrocarpum sensu Wall and Dale 1966, Polysphaeridium zoharyi, and heterotrophic Brigantedinium ssp., cyst of Gymnodinium catenatum and cysts mixture of Protoperidinium. Three local dinoflagellate zones LAX-1 to LAX-3 were established based on the results of the constrained cluster analysis and data ordination; additionally, the middle zone LAX-2 was derived into two subzones, LAX-2a and LAX-2b based on the dynamics of toxic and heterotrophic cysts as well as on the significant changes (probability, P=0.89) in percentages of eutrophic indicators. The total cyst count varied from 106 to 410 dinocysts per slide, with 177 cyst types on average. Dinocyst assemblages are characterized by high values of the dost-depositional degradation index (kt) that varies between 3.6 and 7.6 (averaging 5.4), which is relatively high and is very typical for the areas with selective dinoflagellate cyst preservation that is related to bottom-water oxygen concentrations.

Keywords: reconstruction of palaeoenvironment, harmful algal blooms, anthropogenic influence on coastal zones, South China Sea

Procedia PDF Downloads 53
9 Cenomanian-Turonian Oceanic Anoxic Event, Palynofacies and Optical Kerogen Analysis in Abu Gharadig Basin, Egypt

Authors: Mohamed Ibrahim, Suzan Kholeif

Abstract:

The Cenomanian-Turonian boundary was a ‘greenhouse’ period. The atmosphere at that time was characterized by high CO₂; in addition, there was the widespread deposition of organic-rich sediments anomalously rich in organic carbon. The sediments, palynological, total organic carbon (TOC), stable carbon and oxygen isotopes (δ¹³C, δ¹⁸O, organic) of the Cenomanian-Turonian Bahariya and basal Abu Roash formations at the southern Tethys margin were studied in two deep wells (AG5 and AG-13), Abu Gharadig Oil Field, North Western Desert, Egypt. Some of the marine (dinoflagellate cysts), as well as the terrestrial palynoflora (spores and pollen grains), reveal extinction and origination patterns that are known elsewhere, although other species may be survived across the Cenomanian-Turonian boundary. This implies control of global changes on the palynoflora, i.e., impact of Oceanic Anoxic Event OAE2 (Bonarelli Event), rather than changes in the local environmental conditions. The basal part of the Abu Roach Formation ('G' and 'F' members, late Cenomanian) shows a positive δ ¹³C excursion of the organic fraction. The TOC is generally high between 2.20 and 3.04 % in the basal Abu Roash Formation: shale of 'G' and carbonate of 'F' members, which indicates that these two members are the main Cretaceous source rocks in the Abu Gharadig Basin and have a type I-II kerogen composition. They are distinguished by an abundance of amorphous organic matter AOM and Chlorococcalean algae, mainly Pediastrum and Scenedesmus, along with subordinate dinoflagellate cysts.

Keywords: oceanic anoxic event, cenomanian-turonian, palynofacies, western desert, Egypt

Procedia PDF Downloads 90
8 Dinoflagellate Thecal Plates as a Green Cellulose Source

Authors: Alvin Chun Man Kwok, Wai Sun Chan, Wei Yuan, Joseph Tin Yum Wong

Abstract:

Cellulose, the most abundant biopolymer, is the major constituent of plant and dinoflagellate cell walls. Thecate dinoflagellates, in particular, are renowned for their remarkable capacity to synthesize intricate cellulosic thecal plates (CTPs). Unlike the extracellular two-dimensional structure of plant cell walls, these CTPs are three-dimensional and reside within the cellular structure itself. The deposition of CTPs occurs with remarkable precision, and their arrangement serves as crucial taxonomic markers. It is noteworthy that these plates possess the hardness of wood, despite the absence of lignin. Partial and prolonged hydrolysis of CTPs results in the formation of uniform long bundles and lowdimensional, modular crystalline whiskers. This observation aligns with the consistent nanomechanical properties, suggesting a CTPboard structure. The unique composition and structural characteristics of CTPs distinguish them from other cellulose-based materials in the natural world. Spectroscopic studies using Raman and FTIR methods indicate a clear low crystallinity index, with the OH shift becoming more distinct following SDS treatment. Birefringence imaging confirms the highly organized structure of CTPs, demonstrating varying degrees of anisotropy in different regions, including both seaward and cytosolic passages. The knockdown of a cellulose synthase enzyme in dinoflagellates resulted in severe malformation of CTPs and hindered the life-cycle transition. Unlike certain other microalgal groups, these unique circum-spherical depositions of CTPs were not pre-fabricated and transported "to site," but synthesized within alveolar sacs at the specific site. Our research is particularly focused on unraveling the mechanisms underlying the biodeposition of CTPs and exploring their potential biotechnological applications. Understanding the processes involved in CTP formation can pave the way for harnessing their unique properties for various practical applications. Dinoflagellates play a crucial role as major agents of algal blooms and are also known for producing anti-greenhouse sulfur compounds such as DMS/DMSP, highlighting the significance of CTPs as a carbon-neutral source of cellulose. Grant acknowledgement: Research in the laboratory are supported by GRF16104523 from Research Grant Council to JTYW.

Keywords: cellulosic thecal plates, dinoflagellates, cellulose, cell wall

Procedia PDF Downloads 39
7 Spatial Variability of Phyotoplankton Assemblages during the Intermonsoon in Baler Bay, Outer and Inner Casiguran Sound, Aurora, Fronting Philipine Rise

Authors: Aime P. Lampad-Dela Pena, Rhodora V. Azanza, Cesar L. Villanoy, Ephrime B. Metillo, Aletta T. Yniguez

Abstract:

Phytoplankton community changes in relation to environmental parameters were compared between and within, the three interconnected basins. Phytoplankton samples were collected from thirteen stations of Baler Bay and Casiguran Sound, Aurora last May 2013 by filtering 10 L buckets of surface water and 5 L Niskin samples at 20 meters and at 30 to 40 meters depths through a 20um sieve. Duplicate samples per station were preserved, counted, and identified up to genus level, in order to determine the horizontal and vertical spatial variation of different phytoplankton functional groups during the summer ebb and flood flow. Baler Bay, Outer and Inner Casiguran Sound had a total of 89 genera from four phytoplankton groups: Diatom (62), Dinoflagellate (25), Silicoflagellate (1) and Cyanobacteria (1). Non-toxic diatom Chaetoceros spp. bloom (averaged 2.0 x 105 to 2.73 x 106 cells L⁻¹) co-existed with Bacteriastrum spp. at surface waters in Inner and Outer Casiguran. Pseudonitzschia spp. (1.73 x 106 cells L⁻¹) bloomed at bottom waters of the innermost embayment near Casiguran mangrove estuary. Cyanobacteria Trichodesmium spp. significantly increased during ebb tide at the mid-water layers (20 meters depth) in the three basins (ranged from 6, 900 to 15, 125 filaments L⁻¹), forming another bloom. Gonyaulax spp. - dominated dinoflagellate did not significantly change with depth across the three basins. Overall, diatoms and dinoflagellates community assemblages significantly changed between sites (p < 0.001) while diatoms and cyanobacteria varied within Casiguran outer and inner sites (p < 0.001) only. Tidal fluctuations significantly affected dinoflagellates and diatom groups (p < 0.001) in inner and baler sites. Chlorophyll significantly varied between (KW, p < 0.001) and within each basins (KW, p < 0.05), no tidal influence, with the highest value at inner Casiguran and at deeper waters indicating deep chlorophyll maxima. Aurora’s distinct shelf morphology favoring counterclockwise circulation pattern, advective transport, and continuous stratification of the water column could basically affect the phytoplankton assemblages and water quality of Baler Bay and Casiguran inner and outer basins. Observed spatial phytoplankton community changes with multi-species diatom and cyanobacteria bloom at different water layers of the three inter-connected embayments would be vital for any environmental management initiatives in Aurora.

Keywords: aurora fronting Philippines Rise, intermonsoon, multi-species diatom bloom, spatial variability

Procedia PDF Downloads 109
6 A Holistic View of Microbial Community Dynamics during a Toxic Harmful Algal Bloom

Authors: Shi-Bo Feng, Sheng-Jie Zhang, Jin Zhou

Abstract:

The relationship between microbial diversity and algal bloom has received considerable attention for decades. Microbes undoubtedly affect annual bloom events and impact the physiology of both partners, as well as shape ecosystem diversity. However, knowledge about interactions and network correlations among broader-spectrum microbes that lead to the dynamics in a complete bloom cycle are limited. In this study, pyrosequencing and network approaches simultaneously assessed the associate patterns among bacteria, archaea, and microeukaryotes in surface water and sediments in response to a natural dinoflagellate (Alexandrium sp.) bloom. In surface water, among the bacterial community, Gamma-Proteobacteria and Bacteroidetes dominated in the initial bloom stage, while Alpha-Proteobacteria, Cyanobacteria, and Actinobacteria become the most abundant taxa during the post-stage. In the archaea biosphere, it clustered predominantly with Methanogenic members in the early pre-bloom period while the majority of species identified in the later-bloom stage were ammonia-oxidizing archaea and Halobacteriales. In eukaryotes, dinoflagellate (Alexandrium sp.) was dominated in the onset stage, whereas multiply species (such as microzooplankton, diatom, green algae, and rotifera) coexistence in bloom collapse stag. In sediments, the microbial species biomass and richness are much higher than the water body. Only Flavobacteriales and Rhodobacterales showed a slight response to bloom stages. Unlike the bacteria, there are small fluctuations of archaeal and eukaryotic structure in the sediment. The network analyses among the inter-specific associations show that bacteria (Alteromonadaceae, Oceanospirillaceae, Cryomorphaceae, and Piscirickettsiaceae) and some zooplankton (Mediophyceae, Mamiellophyceae, Dictyochophyceae and Trebouxiophyceae) have a stronger impact on the structuring of phytoplankton communities than archaeal effects. The changes in population were also significantly shaped by water temperature and substrate availability (N & P resources). The results suggest that clades are specialized at different time-periods and that the pre-bloom succession was mainly a bottom-up controlled, and late-bloom period was controlled by top-down patterns. Additionally, phytoplankton and prokaryotic communities correlated better with each other, which indicate interactions among microorganisms are critical in controlling plankton dynamics and fates. Our results supplied a wider view (temporal and spatial scales) to understand the microbial ecological responses and their network association during algal blooming. It gives us a potential multidisciplinary explanation for algal-microbe interaction and helps us beyond the traditional view linked to patterns of algal bloom initiation, development, decline, and biogeochemistry.

Keywords: microbial community, harmful algal bloom, ecological process, network

Procedia PDF Downloads 80
5 Biodeterioration and Biodegradation of Historic Parks of UK by Algae

Authors: Syeda Fatima Manzelat

Abstract:

The present study aims to study the groups of algal genera that are responsible for the biodeterioration, biodegradation, and biological pollution of the structures and features of the two historic parks of the UK. Different sites of Campbell Park and Great Linford Manor Park in Milton Keynes are selected to study the morphological, aesthetic, and physical effects of the algal growth. Specimens and swabs were collected mechanically from selected sites. Algal specimens are preserved in Lugol’s solution and labelled with standard information. Photomicrograph analysis of slides using taxonomic keys and visual observation identified algal species that are homogenously and non-homogenously mixed in the aerial, terrestrial, and aquatic habitats. A qualitative study revealed seven classes of Algae. Most of the algal genera isolated have proven records of potential biodegradation, discoloration, and biological pollution. Chlorophyceae was predominantly represented by eleven genera: Chlorella, Chlorococcum Cladophora, Coenochloris Cylindrocapsa, Microspora, Prasiola, Spirogyra, Trentepholia, Ulothrix and Zygnema. Charophyceae is represented by four genera: Cosmarium, Klebsormidium, Mesotaenium, and Mougeotia. Xanthophyceae with two genera, Tribonema and Vaucheria. Bacillariophyceae (Diatoms) are represented by six genera: Acnanthes, Bacillaria, Fragilaria, Gomphonema, Synedra, and Tabellaria, Dinophyceae with a Dinoflagellate. Rhodophyceae included Bangia and Batrachospermum, Cyanophyceae with five genera, Chroococcus, Gloeocapsa, Scytonema, Stigonema and Oscillatoria. The quantitative analysis by statistical method revealed that Chlorophyceae was the predominant class, with eleven genera isolated from different sites of the two parks. Coenochloris of Chlorophyceae was isolated from thirteen sites during the study, followed by Gloeocapsa of Cyanophyceae, which is isolated from 12 sites. These two algae impart varying shades of green colour on the surfaces on which they form biofilms. Prasiola, Vaucheria, and Trentepholia were isolated only from Great Linford Park. Trentepholia imparted a significant orange colour to the walls and trees of the sites. The compounds present in algae that are responsible for discoloration are the green pigment chlorophyll, orange pigment β-carotene, and yellow pigment quinone. Mesotaenium, Dinoflagellate, Gomphonema, Fragilaria, Tabellaria and two unidentified genera were isolated from Campbell Park only. Largest number of algal genera (25) were isolated from the canal of Campbell Park followed by (21) from the canal at Great Linford Manor Park. The Algae were found to grow on surfaces of walls, wooden fencings, metal sculptures, and railings. The Algae are reported to induce surface erosion, natural weathering, and cracking, leading to technical and mechanical instability and extensive damage to building materials. The algal biofilms secrete different organic acids, which are responsible for biosolubilization and biodeterioration of the building materials. The aquatic algal blooms isolated during the study release toxins which are responsible for allergy, skin rashes, vomiting, diarrhea, fever, muscle spasms, and lung and throat infections. The study identifies the places and locations at the historic sites which need to be paid attention. It provides an insight to the conservation strategies to overcome the negative impacts of bio colonization by algae. Prevention measures by different treatments need to be regularly monitored.

Keywords: algae, biodegradation, historic gardens, UK

Procedia PDF Downloads 13
4 Harmful Algal Blooms in Omani and Arabian Sea and Their Effect on Marine Environment

Authors: Hamed Mohammed Al Gheilani

Abstract:

Red tide, one of the harmful algal blooms (HABs) is a natural ecological phenomenon and often this event is accompanied by severe impacts on coastal resources, local economies, and public health. The occurrence of red tides has become more frequent in Omani waters in recent years. Some of them caused fish kill, damaged fishery resources and mariculture, threatened the marine environment and the osmosis membranes of desalination plants. However, a number of them have been harmless. The most common dinoflagellate Noctiluca scintillans is associated with the red tide events in Omani waters. Toxic species like Karenia selliformis, Prorocentrum arabianum, and Trichodesmium erythraeum have also been reported recently. Although red tides in Oman have been considered a consequence of upwelling in the summer season (May to September), recent phytoplankton outbreaks in Oman are not restricted to summer. Frequent algal blooms have been reported during winter (December to March). HABs may have contributed to hypoxia and/or other negative ecological impacts. The effects of HABs on desalination plan were increased in last three years, by blooms of Cochlodinium, noctiluca species, and blooms of jellyfish. Most of these blooms were affected Al Batinah and Muscat coast. These effects include millions of Omani Rials and several shutdowns of desalination plans during these years.

Keywords: red tide, environment, hypoxia, noctiluca

Procedia PDF Downloads 401
3 Palynology of the Cretaceous Deposits of the Southeast Sirt Basin, Libya

Authors: Khaled Mahmud Gaddah

Abstract:

Cretaceous deposits in the southeast of the Sirt Basin of Libya occur in several grabens that formed during a major phase of rifting related to the opening of the Tethys. They include continental deposits of Early Cretaceous age that belong to the Nubian Formation and marginal to fully marine deposits of Late Cretaceous age that belong to the Lidam Formation and transitional beds. The sequence was extensively sampled from twenty-two boreholes and palynologically analysed. Much of the sequence is barren. However, subordinate shales in all formations yield diverse assemblages of poorly to well preserved and thermally middle to post mature palynomorphs. Most of the assemblages contain non-marine palynomorphs (spores, pollen, and freshwater algae), although some contain rare marine elements (dinoflagellate cysts and acritarchs). The palynofloras enabled the recognition of six assemblage zones of the late Barremian-Turonian age based on the dominant and base/top occurrences of stratigraphically useful palynomorphs: AI (Afropollisspp.-Aequitriraditesspinulosus) of late Barremian age; AIIa (Scenedesmusbifidus-S. sp.) of late Barremian?-early Aptian age; AIIb (Afropollisoperculatus-A. zonatus) of Aptian age; AIII (Crybelosporitespannuceus-Afropollisjardinus) of early Albian age; AIV (Subtilisphaera sp.-Lophosphaeridiumspp.) of Cenomanian-?Turonian age; AIV (Pediastrumbifidites-Leiosphaeridiaspp.) of Cenomanian?-Turonian age. These assemblages are comparable to others from Northern Gondwana (particularly from Libya and Egypt) and correspond to established Cretaceous palynofloral provinces. Palynofacies analysis is used to interpret the depositional environments, and five palynofacies types are recognised that reflect increasing marine influence up section.

Keywords: palynology, palynomorphs, palynofacies, tethys, sirt basin

Procedia PDF Downloads 61
2 Appearance of Ciguatoxin Fish in Atlantic Europe Waters

Authors: J. Bravo, F. Cabrera Suárez, B. Vega, L. Román, M. Martel, F. Acosta

Abstract:

Ciguatera fish poisoning (CFP) is the most common non-bacterial intoxication in the world caused by ingestion of fish with bio-accumulated ciguatoxins (CTXs). It is typical in tropical and subtropical areas, mainly affecting the Caribbean Sea, Polynesia and other areas in the Pacific and Indian Oceans. Interest in Europe by the CFP is increasing in recent years as more and more cases in European hospitals are appearing, usually by people who have consumed ciguatoxin imported fish or have travelled to areas of risk for this poisoning. Since 2004 a series of poisonings raised the question of a possible occurrence of ciguatoxin in Europe, especially in the area of Macaronesia in the East Atlantic temperate zone. Furthermore, some studies have identified the presence of Gambierdiscus spp. in waters surrounding the Canary Islands and Madeira, a toxic dinoflagellate related to this poisoning. The toxin accumulates and concentrates through the food chain and affects to the end of the chain, the human consumer. Fish were collected from the Canary Islands waters and the toxin has been extracted and purified by using acetone and liquid/liquid partition in order to eliminate the excess of fatty acids that may interfere with the detection of the toxin. The fish extracts were inoculated in Neuroblastoma (neuro-2a) cells. After 24-h cell viability was used as an endpoint for cytotoxic effects measurement. Since 2011 our laboratory is collecting data for species such Seriola spp., Epinephelus spp., Makaira spp., Pomatomus spp., Xiphias spp., and Acantocybium spp., from all islands and including the sports fishing and professional activities, we obtained a 8% of fish that have ciguatoxin in their muscle. With these results, we conclude that the island where fishing and fish size affects the probability of catching a fish with the ciguatoxin.

Keywords: Canary Islands, ciguatera fish poisoning, ciguatoxin, Europe

Procedia PDF Downloads 311
1 The Development of Noctiluca scintillans Algal Bloom in Coastal Waters of Muscat, Sulanate of Oman

Authors: Aysha Al Sha'aibi

Abstract:

Algal blooms of the dinoflagellate species Noctiluca scintillans became frequent events in Omani waters. The current study aims at elucidating the abundance, size variation and observations on the feeding mechanism performed by this species during the winter bloom. An attempt was made, to relate observed biological parameters of the Noctiluca population to environmental factors. Field studies spanned the period from December 2014 to April 2015. Samples were collected from Bandar Rawdah (Muscat region) by Bongo nets, twice per week, from the surface and the integrated upper mixed layer. The measured environmental variables were: temperature, salinity, dissolved oxygen, chlorophyll a, turbidity, nitrite, phosphate, wind speed and rainfall. During the winter bloom (from December 2014 through February 2015), the abundance exhibited the highest concentration on 17 February (640.24×106 cell.L-1) in oblique samples and 83.9x103 cell.L-1 in surface samples, with a subsequent decline up to the end of April. The average number of food vacuoles inside Noctiluca cells was 1.5 per cell; the percentage of feeding Noctiluca compared to the entire population varied from 0.01% to 0.03%. Both the surface area of the Noctiluca symbionts (Pedinomonas noctilucae) and cell diameter were maximal in December. In oblique samples the highest average cell diameter and the surface area of symbiont algae were 751.7 µm and 179.2x103 µm2 respectively. In surface samples, highest average cell diameter and the surface area of symbionts were 760 µm and 284.05x103 µm2 respectively. No significant correlations were detected between Noctiluca’s biological parameters and environmental variables except for the correlation between cell diameter and chlorophyll a, also between symbiotic algae surface area and chlorophyll a. The high correlation of chlorophyll a was as a reason of endosymbiotic algae Pedinomonas noctilucae and green Noctiluca enhanced chlorophyll during bloom. All correlations among biological parameters were significant; they are perhaps one of major factors that mediating high growth rates, generating millions of cell per liter in a short time range. The results gained from this study will provide a beneficial background for understanding deeply the development of coastal algal blooms of Noctiluca scintillans. Moreover, results could be used in different applications related to marine environment.

Keywords: abundance, feeding activities, Noctiluca scintillans, Oman

Procedia PDF Downloads 400