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Analysis of Species Diversity in Zooplankton Using Environmental DNA Sequencing Techniques

In light of global environmental changes and the increasing interest in marine species diversity, studying species diversity from marine ecosystems has become vital. This article focuses on research conducted in the Ulleng Basin in South Korea, known for its rich diversity of zooplankton species. By integrating two innovative techniques, bulk DNA analysis and environmental DNA (eDNA) using high-throughput sequencing, the study reveals that the number of species discovered exceeds 350, which represents three to six times more than what has been recognized through traditional methods. Here, we will review the importance of these findings and how the use of non-invasive methods rekindles hope in expanding our understanding of marine species diversity and ecosystem dynamics, aiding in the improvement of conservation efforts and the management of marine resources.

Diversity of Microanimal Species and Its Importance in the Marine Ecosystem

Microanimal species, known as zooplankton, are considered fundamental groups within the marine ecological balance. They play a vital role in the marine food chain and indirectly affect fishery resources and water quality. Studying the diversity of zooplankton species is an important indicator of changes in the marine ecosystem, as these organisms respond quickly to environmental changes such as rising temperatures or alterations in water chemistry. By monitoring these species, scientists can assess the health of the ecosystem and better guide environmental policies.

Historical data indicate that zooplankton play a pivotal role in the biological cycle of carbon and nitrogen elements. For instance, zooplankton help migrate carbon from the surface to the depths of the ocean – a process known as the carbon cycle, which contributes to reducing carbon dioxide levels in the atmosphere. Furthermore, zooplankton are an integral part of marine food chains, serving as vital food for many marine species.

However, scientists face significant challenges in identifying zooplankton species diversity using traditional methods, such as morphological examination, which relies on the morphological characteristics of organisms. These processes require advanced expertise from specialized scientists and can be time-consuming. This process is particularly challenging when morphological traits are unclear or similar between species.

DNA Analysis Technology and Its Impact on Biodiversity Studies

Biological sciences have advanced significantly with the introduction of DNA analysis techniques in studying biodiversity. DNA sequencing “barcoding” enhances the ability to identify species quickly and accurately, as this analysis relies on using a specific segment of DNA that can distinguish between different species. The use of metabarcoding, an advanced DNA analysis technique, is one of the major developments in this field, allowing for the analysis of mixed samples containing multiple species groups.

For example, metabarcoding has been used in studying a zooplankton sample by sequencing DNA from samples collected from the sea floor. Through this technique, researchers were able to identify 350 zooplankton species – a number three to six times larger than the species identified using traditional methods. This indicates the effectiveness of more comprehensive sampling methods that include both eDNA and bulk DNA to ensure an accurate picture of species diversity.

These modern techniques also seek to overcome the drawbacks of traditional methods such as habitat destruction and sampling issues. For example, traditional nets used for collecting zooplankton may lead to the destruction of the natural habitats of marine organisms. In contrast, environmental DNA analysis allows for data collection on species that are difficult to capture from the environment, contributing to a deeper understanding of species relationships and their geographic distribution.

Analysis

Environmental and Benefits of Using Environmental DNA

The technique of environmental DNA (eDNA) is considered an innovative and non-invasive method for gathering information about biodiversity in marine environments. Environmental DNA is the genetic material released into the environment by living organisms, serving as an effective means to assess biodiversity without the need to collect physical samples. This has several benefits, including the ability to identify species that may be difficult or impossible to collect using traditional methods.

Through rapid testing, the environmental DNA technique can detect the presence of absent species using small amounts of DNA preserved in the environment. This can have significant implications for environmental protection policies, as this data can be used to identify areas rich in biodiversity.

The outputs of this technique go beyond mere species identification, as it can also provide deeper insights into the ecological relationships among species and their life cycles. For example, if environmental DNA test results show unexpected increases in certain species of zooplankton, these findings might indicate environmental changes that warrant investigation.

By utilizing environmental DNA analysis, researchers can gain a comprehensive picture of changes in the marine environment and identify any potential threats facing the ecosystem. This is considered a vital tool in addressing environmental challenges such as climate change and ocean pollution, providing valuable information that enhances the effectiveness of conservation and environmental management programs.

Methods for Extracting and Analyzing DNA

DNA extraction is a vital process in scientific studies and biological research. In this process, modern techniques and equipment are used to ensure the integrity of samples and the quality of the extracted DNA. Centrifugation has been utilized to separate DNA due to its high capability to remove impurities, such as residual ethanol, which can affect DNA analysis. Additionally, steel beads were used to mix the samples thoroughly, ensuring cell lysis and DNA release. The process typically lasts for several minutes at a specific frequency, facilitating the subsequent analysis.

Once high-quality DNA is obtained, advanced techniques such as polymerase chain reaction (PCR) are employed, playing a crucial role in amplifying targeted regions of DNA. At this step, specialized primers are used that bind to specific points in the DNA sequence, increasing the amount of genetic material necessary for further analyses. Optimal thermal conditions are utilized to ensure the success of the reaction, resulting in the creation of a large number of copies of the target gene, facilitating identification and analysis.

Sequencing Techniques and Biological Analysis

Sequencing techniques are an essential part of molecular biology, allowing researchers to examine biodiversity and distinguish between different species. The Illumina MiSeq sequencing platform was used to gather genomic data from the analyzed samples. After conducting the sequencing analysis, biostatistical techniques are applied to analyze the data and support research objectives. These techniques include programs like Qiime2, which enable scientists to manage biological data, filter taxa, and subsequently analyze them.

Statistical tools such as geometric mean and other measures are utilized to estimate biodiversity, ensuring a comprehensive representation of different species. For instance, the Shannon index was calculated to classify species based on their abundance and diversity, a common metric used to analyze ecological diversity. The Shannon index provides a broad view of the relative distribution of different species in the studied sample.

Analysis Results and Biodiversity

The results obtained from the analysis provided deep insights into the biodiversity of different species in the studied area. A total of 350 species of organisms were identified, distributed across 12 main groups, with the arthropod group (Arthropoda) being the richest in terms of the number of species present. This diversity indicates the stability of the current ecosystem and also provides important indicators of the overall health of the marine environment.

Considering

To numbers and data, it appears that the Copepoda group was the most diverse, containing 96 species. These results reflect the importance of these species in marine food webs, and may also indicate specific environmental activities that may affect their diversity and abundance. Various types of crustaceans were identified at all studied stations, reflecting their wide presence in the marine system.

Overall, the results of species analysis highlight the environmental impacts and molecular growth of marine life, as the data extracted through sequencing indicate not only the presence of living organisms but also their complex interactions within the ecosystem. These conclusions play a vital role in conservation strategies and maintaining biodiversity in the oceans.

Challenges and Opportunities in Studying Biodiversity

Despite significant technological advances in the fields of molecular biology and sequencing, biodiversity studies still face many challenges. Some of these challenges relate to maintaining the quality of the samples used, the ability to distinguish between genetically similar species, and ensuring the accuracy of the statistical analyses used. In some cases, differences in sampling methods or data processing can lead to contradictory results, requiring special attention to each stage of research.

Nevertheless, these challenges present new opportunities for research and discovery. Innovations in sequencing technology are accelerating, increasing the ability to generate detailed and accurate data. Developments in software such as Qiime2 and R represent powerful tools that help researchers process and analyze their data more effectively. Additionally, the new data being collected provides scientists with the opportunity to reassess old hypotheses about biodiversity and species presence.

Ultimately, while studying biodiversity represents a challenge that requires a precise and innovative approach, the benefits gained from learning more deeply about the aquatic environment represent an opportunity not to be missed for improving biodiversity conservation strategies. Through continuous work and collaboration among scientists, a deeper understanding of a sustainable and healthy world can be developed.

Biodiversity in Oceans

Biodiversity in oceans represents one of the most important elements of marine ecosystems, reflecting the variety of living species that inhabit these aquatic environments. The importance of this diversity lies in its role in maintaining ecological balance, as it contributes to the sustainability of marine life and ensures the functions of the ecosystem. The species in the oceans vary from microscopic and invisible organisms to massive marine creatures, such as marine mammals. This diversity is influenced by several factors, including climate change and human activities such as overfishing and pollution.

Research indicates that the oceans host an estimated 230,000 species of living organisms, but the actual number is expected to be much higher. For instance, some endangered species due to human activities include marine organisms such as fish, invertebrates, and marine plants. This issue raises concerns about the ability of marine ecosystems to adapt and survive in the face of increasing challenges. Moreover, studies have shown that biodiversity enhances ecosystem resilience to changes and increases their flexibility.

To better understand biodiversity, scientists are increasingly relying on techniques such as DNA sequencing and genetic technologies to identify different species. These techniques provide accurate information about biodiversity, facilitating conservation and management of marine resources more effectively. For example, gene sequencing has been used to determine the relationships between marine species and the environments they inhabit, providing new insights into how environmental changes affect diversity in the oceans.

Methods

Identification of Marine Species

Traditional methods of identifying marine species are limited in their ability to recognize biodiversity, prompting scientists to turn to modern techniques such as DNA analysis. DNA analysis is a powerful tool that allows researchers to study biodiversity with high precision, enabling them to identify species by examining sequences of DNA. Techniques such as Metabarcoding highlight the capability of this method to detect a wide range of species simultaneously.

Many studies have applied DNA analysis techniques in marine environments, and the results have often been quite remarkable. For example, a recent study showed a significant increase in the number of species discovered using Metabarcoding compared to traditional methods. Researchers were able to identify over 350 species in a specific area using DNA, demonstrating the efficiency of this method in the field of environmental research.

The benefits of DNA techniques go beyond species identification; they also serve as a revolutionary tool for monitoring changes in biodiversity over time. By collecting and analyzing samples from marine environments, scientists can track new species extinctions or the emergence of new species that may result from environmental changes. This makes them essential in conservation research and resource management.

Threats to Biodiversity in the Oceans

The oceans face increasing threats from human activity, making biodiversity vulnerable. Studies indicate that climate change, pollution, and overfishing are among the main threats affecting marine species. Climate change impacts marine ecosystems by raising ocean temperatures, leading to coral reef degradation and changes in marine food webs.

Statistics show that nearly 30% of coral reefs around the world have already deteriorated, threatening species that depend on these habitats. Furthermore, research has proven that chemical pollution, such as plastic pollutants and heavy metals, directly affects the health of marine organisms. These pollutants accumulate in the food chain, threatening not only marine life but also the health of humans who consume seafood.

Moreover, overfishing negatively impacts biodiversity in the oceans, leading to significant declines in marine species populations. Some species are being unsustainably depleted, necessitating more effective assurances in fishery management and sustainability of fish stocks.

Conservation Strategies and Necessary Measures

Addressing the threats to biodiversity in the oceans requires the implementation of comprehensive conservation strategies. It is essential to enhance protective measures for endangered species and conservation efforts at the ecosystem level as a whole. These strategies include the establishment of marine protected areas, where destructive activities such as fishing and seabed mining are prohibited, allowing marine ecology to recover and regenerate.

Additionally, local communities and stakeholders must promote awareness of biodiversity and its importance. Education and outreach play a crucial role in fostering a conservation culture among individuals, helping to motivate people to engage in conservation activities and make sustainable decisions regarding the use of marine resources.

There is also an urgent need to enhance research and studies on biodiversity in the oceans, in addition to applying modern technology such as DNA analysis to track species and monitor changes. Accurate data is essential for understanding ecological patterns, and for strategic planning aimed at preserving biodiversity. International exchange of knowledge and experiences can assist many countries in developing effective policies for ocean protection.

Species Identification Using Molecular DNA Analysis

Contributes
DNA metabarcoding has significantly advanced the identification of new or rarely observed species. This technique relies on matching DNA sequences with a reference database, such as the MZG database, where a sequence is considered a match when it shows 99% alignment with sequences in the database. The reliability of metabarcoding is closely linked to the scope and quality of the reference database, making continuous updates to these reference libraries essential for enhancing the accuracy and objectivity of research results. By aggregating information over time, these updates can reduce the number of unknown sequences and also aid in the discovery of species that have not yet been described, particularly in groups that have not been well studied.

In this context, metabarcoding has demonstrated exceptional capability in species identification, with 99% of the amplicon sequence variants (ASVs) defined at the species or genus level, outperforming the recognition rates achieved by traditional morphology-based methods. However, significant challenges remain in species identification, particularly in certain groups such as amphipods and viruses, as well as developmental stages like the larvae of copepods. Cryptic species create particular challenges for species identification in groups such as Bivalvia, Hydrozoa, Polychaeta, and Scyphozoa, representing an additional hurdle for scientists.

Problems related to species identification through morphological examination stem from the limited availability of information on species classification, variations in species records themselves, as well as the significant impact of developmental stages and any damage to samples. In contrast, metabarcoding data provides updated information accessible in real-time through resources like MZG, GenBank, BOLD, and SILVA. This technology has the potential to overcome the challenges facing traditional species identification methods, making it an extremely powerful tool in ecology.

Methods of Data Collection from Environmental DNA and Total DNA

Studies employ various methods to collect data on biodiversity, including environmental DNA (eDNA) analysis and total DNA from net samples. These methods contribute to a comprehensive picture of genetic diversity in marine systems. In one study, data were collected from 119 species found in DNA samples, focusing on taxonomic groups like Copepoda, Actinopterygii, and Hydrozoa. By using both sampling methods, a total of 160 species of total DNA and 71 species of eDNA were discovered, reflecting the importance of both approaches.

Issues affecting species diversity can be attributed to the type and size of nets used for sampling. Research shows that species diversity can be significantly impacted by the type of nets used; for example, traditional nets with larger mesh sizes (200-300 micrometers) lead to underestimates of species diversity among very small species that are not effectively collected. For this reason, ultra-fine nets are necessary for gathering comprehensive information.

Other factors affecting sampling include swimming ability. Species that swim well, such as fish, are difficult to collect using traditional nets, while gelatinous species like jellyfish can incur damage during collection. Therefore, environmental DNA analysis comes as an effective solution to these problems, as it analyzes DNA fragments taken from the environment, enabling the detection of a diverse range of species, from nano-plankton (<2 micrometers) to mega-plankton (>2 centimeters).

Differences

On Biodiversity According to Sampling Methods

During marine surveys using both total DNA and environmental DNA, the results showed significant differences in species compositions and diversity. For example, there were clear differences among Taxa groups, including Copepoda, Actinopterygii, Gastropoda, and Hydrozoa. It was observed that some groups, such as Copepoda and Gastropoda, were more prominently identified in total DNA, while species like Actinopterygii and Cephalopoda were more distinctly represented in environmental DNA analyses. This is due to the high swimming speed of these species, making their collection using traditional nets nearly impossible.

In contrast, the study provided results similar to many previous studies, showing that living species in aquatic categories like Scyphozoa were better detected through environmental DNA analysis. This is due to their fragility and difficulty in collection using nets, which increased the effectiveness of methods based on environmental DNA analysis in revealing diverse species. This difference in species identification reflects each method’s reliance on its specific advantages to overcome the limitations of traditional methods, enabling scientists to gain deeper insights into the biodiversity of the marine environment.

The General Importance of Genetic Studies in Biodiversity

Genetic studies, such as polymorphic DNA analysis, reflect the importance of Robus in improving species classification and research accuracy in marine environments. These methods allow the discovery of over 350 different marine species, with identification accuracy exceeding 99%. These results confirm the exceptional ability of DNA techniques to enhance scientific understanding of biodiversity compared to traditional methods. Furthermore, the significant differences in species diversity according to sampling methods indicate that integrating different analytical approaches is essential for obtaining a more accurate and comprehensive picture of biodiversity. These advancements are crucial for understanding marine ecosystems’ responses to climate changes and other environmental considerations.

Thus, DNA analysis represents a key tool in biodiversity conservation, highlighting the need for continued development and innovation in biological research techniques, as well as the importance of sustainable research to improve this field. It also encourages a rapid response to artificial and natural environmental changes affecting marine ecosystems. By using DNA as a means of identifying species, the information gathered with environmental data is vital for mapping biodiversity and ensuring ecosystem sustainability.

Identifying Fish Species Using Metabarcoding Techniques

Metabarcoding technology is one of the modern and effective tools in marine biology, used to identify fish species through DNA analysis. This process involves collecting samples from aquatic environments and analyzing the environmental DNA (eDNA) present in them. This technique allows researchers to study biodiversity and understand the ecological balance of marine resources dynamically. The main difference between traditional data collection methods and metabarcoding is that the latter relies on examining the DNA left by organisms in their environment, rather than directly extracting these organisms.

For example, in a study conducted off the western shores of Florida, metabarcoding was used to identify fish eggs in the waters. Researchers detected high concentrations of eDNA for all studied species, demonstrating the effectiveness of this method in identifying species even at early life stages. This means that metabarcoding is not only a good tool for scientific studies but also beneficial for managing marine fisheries and ensuring their sustainability.

Moreover, metabarcoding represents a solution to multiple issues faced by marine research, such as the difficulty of collecting physical samples of marine organisms due to their small size or environmental conditions. For instance, collecting eggs or larvae from the depths of the ocean can be a significant challenge, while metabarcoding allows data collection from any point in the water without the need for expensive tools. This enables scientists to obtain accurate information about a wide range of species in a short period.

Challenges

The Risks Associated with Metabarcoding Techniques

While metabarcoding technology offers many benefits, there are challenges and risks that must be taken into account. Among the most significant risks is the accuracy of reading the data generated by this technique. For example, there may be instances of DNA overlap between species, which can lead to difficulties in accurately identifying some species. Additionally, the effectiveness of metabarcoding heavily depends on the quality of the sample collected and the environmental conditions during DNA collection.

Furthermore, varying environmental conditions, such as temperature or nutrient concentration, can lead to differences in DNA concentrations, affecting the results. Other challenges include selecting the appropriate comparison for the data and references, as there are many species that have not yet been documented in DNA databases. Therefore, developing comprehensive global databases is essential to improve the effectiveness of these techniques.

However, researchers remain optimistic about the future of metabarcoding. Through the development and improvement of technology, and support for ongoing research, there is great hope in overcoming these challenges and achieving accurate and comprehensive results that can make a difference in understanding marine ecosystems. Additionally, collaboration between research stations, universities, and government institutions will help manage data collection and analysis more efficiently.

Applications of Metabarcoding in Biodiversity Conservation

Metabarcoding technology is considered an important tool for biodiversity conservation. It serves as an effective means to monitor and identify endangered species or species that need additional protection. For example, by collecting and analyzing eDNA from various locations, it is possible to identify the presence of certain threatened species and understand the extent of human activities affecting them.

Non-governmental organizations and environmental centers utilize metabarcoding in their projects to assess the status of ecological communities. For instance, this technique can be used to investigate the impact of climate change on marine species. By monitoring changes in species diversity at a specific location over time, researchers can determine how ecosystems are affected by climate change or human activities.

Furthermore, this technique can assist in improving fisheries management strategies. By identifying dominant and threatened species based on the data generated from metabarcoding, local authorities can develop sustainable policies that reflect the current state of marine resources. Thus, these strategies can contribute to enhancing sustainability and reducing negative impacts on the marine environment.

In fragile ocean environments, such as areas affected by fishing activities and global warming, the use of metabarcoding becomes a crucial tool for monitoring the overall health of ecosystems. Therefore, promoting these technologies is an important part of future environmental protection and sustainability strategies, contributing to the preservation of biodiversity in the oceans.

The Importance of Plankton Diversity in Marine Ecosystems

Marine plankton, especially zooplankton, serves as a fundamental indicator of changes occurring in marine ecosystems. Studies have shown that marine plankton responds sensitively to environmental changes, making it an effective tool for assessing the health of marine ecosystems. Zooplankton serves as a bioindicator for analyzing changes in their environments, contributing to a deeper understanding of biodiversity and the stability of these systems.

Plankton also plays a vital role in carbon and nitrogen cycling, as well as in energy transfer within marine food webs. When marine animals such as fish feed on plankton, they contribute to providing the necessary energy for marine life and promoting the fishing sector. Understanding the biodiversity of plankton requires precision in identification and classification, reflecting significant challenges dependent on the sampling methods used and environmental observation.

Methods

Species Identification and Barriers in Phytoplankton Biodiversity

In the field of scientific research, several methods are used to identify species and catalog the biodiversity of phytoplankton. The traditional method relies on the morphological characteristics of phytoplankton, but this method can be complex and labor-intensive, requiring extensive expertise from taxonomic specialists. Morphological features often overlap across different species, making recognition a challenge, especially when it comes to phytoplankton at various life stages.

To address this issue, genetic barcoding techniques have emerged as a new tool aimed at precise species identification. This technique is based on DNA sequencing, which can enhance the chances of accurate species identification by comparing genetic data. However, when dealing with environmental research that requires rapid analysis of large sample groups, relying solely on DNA analysis has proven inadequate.

This is where the metabarcode technique comes into play effectively, as it excels in analyzing mixed samples containing multiple species. By analyzing genes isolated from more than one species in the same sample, scientists can obtain a precise picture of the phytoplankton biodiversity in a specific geographic area. In these contexts, researchers can identify hidden species and estimate biodiversity more effectively.

The Impact of Sampling Methods on Phytoplankton Biodiversity

The sampling method is a critical factor in studying phytoplankton diversity, as it affects the quality of the samples collected. Samples are often extracted using phytoplankton nets; however, such techniques have limitations, including high costs and time consumption. The size of the mesh in the nets can also determine the type of phytoplankton collected, which may lead to bias in the data.

Additionally, challenges arise in distinguishing between benthic phytoplankton and planktonic organisms. These types of phytoplankton, such as annelids, often require specialized collection techniques. Therefore, to ensure the best representation of phytoplankton diversity, a combination of various sampling methods must be employed. Such strategies can help universities and research centers achieve greater diversity and uncover species that may be missed in traditional studies.

Future Perspectives on Biodiversity Using Metabarcoding

As metabarcoding techniques advance, scientific studies face new opportunities and barriers. Although the effectiveness of these techniques in enhancing our understanding of phytoplankton biodiversity is evident, they also require improvements in data collection and analysis methods. The future may witness the extensive use of these technologies in various marine environments, from open oceans to more complex coastal areas.

The future also necessitates a greater awareness of the importance of biodiversity and the impact of climate change and human activities on ecosystems. Data collected through metabarcoding can serve as a foundation for developing effective strategies for managing and protecting marine resources. By focusing on the study of phytoplankton biodiversity, we can enhance conservation efforts and ensure sustainability for the future.

Understanding Environmental DNA (eDNA)

Environmental DNA (eDNA) represents a type of DNA that can be extracted from various environments such as seawater and soil. This type of DNA comprises the genetic data found in the remains of organic materials such as feces, saliva, urine, and skin cells released by living organisms. This method is highly effective for detecting species that cannot be collected using traditional nets due to size or swimming ability.

The importance of using eDNA in evaluating biodiversity is increasing, especially in marine ecosystems. eDNA techniques rely on species identification through complex tools like DNA sequencing, allowing researchers to classify marine organisms more accurately without actual interference in the environment. Such analyses can provide a clearer picture of biodiversity, especially in areas that are difficult to access or present significant environmental challenges.

Creating
Modern technology that relies on eDNA analysis is a qualitative leap in the way we understand marine life. For example, by using DNA sequencing analysis, scientists can identify species of organisms from water samples without the need to physically collect them. This reflects a significant improvement in the accuracy of environmental studies and their ability to report biodiversity in more eco-friendly ways.

eDNA analysis requires advanced skills in genetics and necessitates the use of complex technological devices to ensure that the resulting outcomes are accurate and based on strong scientific foundations. Furthermore, previous studies have shown that eDNA can reveal rare species that may not be recorded using traditional methods, making this type of analysis essential in species conservation programs.

Traditional Data Collection Strategies vs. eDNA

Sampling through techniques such as traditional nets is one of the well-known methods in marine environment research. However, research has shown that these methods can lead to bias in results, as nets only attract larger or more actively swimming organisms, leaving out many other species that live on the ocean floor or in deeper waters unrecorded.

In contrast, eDNA techniques provide a more comprehensive way to gather information about marine species. By collecting water samples and analyzing the contained DNA, the species of organisms can be analyzed without harming the environment or using the two methods, making this approach more ethical and efficient. This process is non-intrusive, meaning it does not affect the marine environment as traditional fishing techniques do.

There are many examples that highlight the effectiveness of eDNA. In the Olleong Basin area of the Korean Peninsula, samples were collected to extract DNA from the waters and were used to determine species diversity. In these studies, eDNA provided results that were significantly greater compared to traditional sampling techniques. Thus, species that were previously unknown or suspected of existence became apparent, helping to paint a more accurate picture of biodiversity in that environment.

Many countries have utilized eDNA in biodiversity studies, especially in those regions where natural resources are threatened. This demonstrates how eDNA can be used as a powerful tool not only in scientific research but also in efforts to conserve environmental models. The benefits of eDNA extend beyond environmental data to coordinate species conservation policies and sustainability.

Practical Steps for Collecting and Analyzing eDNA

The steps for collecting and analyzing eDNA involve several stages starting with the collection of samples from targeted environments. In a specific case, samples were collected from the Olleong Basin at various sites and depths over a specified period. The equipment was carefully prepared to avoid contamination between samples, and precise tools were used to professionally collect the samples.

After collecting the samples, the DNA is extracted using specialized kits. For example, the TIANamp Marine Animals DNA Kit can be used to extract DNA from various samples such as eDNA and bulk DNA. This stage requires attention to detail and how to process the samples in a way that minimizes errors.

The next stage proceeds to the polymerase chain reaction (PCR) process where a specific part of the DNA is targeted and amplified for analysis. This stage is critical to ensure that the resulting outcomes accurately reflect the diversity of the targeted species. These steps may involve using techniques such as mass spectrometry to identify the DNA sequences and highlight genetic patterns.

To ensure reliable and accurate results, the data analysis process is conducted through bioinformatics programs like Qiime2, which allow for the understanding of genetic patterns and biological distribution. The data obtained from this analysis is essential for understanding the relationship between different species and their densities in the marine environment.

Importance

Study of Biodiversity and Marine Life Systems

The study of biodiversity and marine life systems is vital for ensuring the health of ecosystems. Biodiversity contributes to enhancing environmental resilience and the availability of life resources. Researching species diversity significantly impacts understanding how these systems function and are organized.

By studying and conserving biodiversity in marine environments, effective strategies can be developed for sustainable development. Species diversity is an indicator of ecosystem health, and when this diversity declines, other ecosystems are affected. For example, a lack of marine species disrupts the food chain, undermining how the ecosystem functions as a whole.

Sustainable conservation strategies require the use of modern science and advanced technologies such as eDNA. Techniques like metafile and genetic analysis contribute to exploring undiscovered species and improving responses to environmental threats like overfishing and climate change.

Issues related to biodiversity represent global challenges that require collaborative efforts between countries and researchers. Studies also provide a deeper understanding of environmental needs and guide political decision-making more effectively, thereby achieving the conservation of marine ecosystems and biodiversity.

Analysis of Amplicon Sequencing Data

High-throughput sequencing technology resulting from nucleic acid sequencing is used to identify the microbial species present in environmental samples. In this study, amplicon sequencing data was analyzed using the Qiime 2 blocks, where amplicon sequence variants (ASVs) that matched with a 99% sequence identity to the MZG database were identified. ASVs that did not meet this criterion were excluded from the analysis, as these bacteria, fungi, and other microbes were present in the data. To enhance the accuracy of the analysis and reduce the impact of cross-contamination, ASVs below the read threshold of 10 were removed from the ASV table. The read data were converted to the WoRMS database for species naming, enhancing the reliability of the analyses.

The read depth for each species was studied, revealing that eDNA-based samples were much lower than those based on bulk DNA, indicating that eDNA may reflect the environmental species diversity but with fewer resources. The process was carried out using multiple experiments across different tools, including statistical studies, data formatting charts, and advanced statistical tools such as NMDS and ANOSIM. The results were significant in understanding species diversity and distribution.

Species Identification and Environmental Composition Analysis

During the analysis, over 23 million reads were obtained, highlighting the richness of the available data. Across 17 bulk DNA samples and 71 eDNA samples, a total of 350 species from 12 different phyla were identified. The most diverse phylum was Arthropoda, which contained 152 species, representing about 43.4% of the identified species. This indicates the diversity of marine environments and their ability to support a wide range of life. The second most common phylum was Chordata, which included 77 species.

This diversity is distributed across multiple groups, including copepods, reflecting the importance of these species in marine food webs. It was noted that the species identified in the analysis showed variable distribution across different stations, reflecting the balance of ecosystems relying on surrounding environmental factors. The data also revealed fluctuations in the population composition between bulk DNA and eDNA species, opening new avenues for better understanding environmental dynamics.

Comparison Between Bulk DNA and eDNA in Biodiversity

Comparative analyses revealed 279 species identified in bulk DNA samples, compared to 190 species identified in eDNA samples. Although the number of species discovered in each type of sample appears distinct, the data suggest that there is a complex overlap among the species. Analyses identified that 119 species, or about 34% of the species, were discovered in both types of samples, with crabs and members of the Chordata phylum being among the groups that were frequently present across the samples.

The analysis shows

The powerful effect of environmental DNA (eDNA) is that it can capture species that are not easily detectable using traditional analysis methods. eDNA analysis is a valuable tool for environmental research, given its ability to detect rare or threatened species through simple water sampling. This advancement enhances the reliance of environmentalists on eDNA as an integrated option for monitoring biodiversity and conserving ecosystems.

Biodiversity Indicators and Statistical Assessment

Indicators such as the Shannon Index have been used to calibrate species diversity, as these metrics are essential for measuring species richness and diversity. The subsequent data analysis process also represents flexibility and comparative insights between different ecosystems. The Wilcoxon test was used to determine the statistical differences between various sample conditions. The results showed significant discrepancies in diversity among the collected samples, indicating the diverse impacts of environmental factors on fossil biodiversity.

Furthermore, NMDS and ANOSIM analyses revealed clear differences in environmental composition among the samples, contributing to a broader understanding of how marine systems respond to environmental changes. These findings detailed how living organisms are affected by environmental changes and highlighted the importance of applying advanced statistical methods in environmental research. Enhancing these analyses will help environmentalists and specialists anticipate and understand changes that may occur in biodiversity due to global challenges such as climate change and pollution.

Species Diversity in Aquatic Ecosystems

Species diversity in aquatic ecosystems is a significant subject in environmental studies. Research focuses on measuring the species present in a specific area and understanding the relationships among species and mechanisms of interaction within the ecological community. In the Ulleung Basin, 350 marine species were identified, reflecting the richness of this ecosystem. Researchers utilized DNA technologies to identify species, which allowed for the detection of species that were previously difficult to identify using traditional recognition methods.

Each group of living organisms has its unique characteristics, with the highest frequency of species occurrence identified in some categories like Copepoda, Gastropoda, Actinopterygii, and Malacostraca. Studies have confirmed that modern techniques such as DNA metabarcoding enhance research effectiveness, as they managed to identify 99% of species at the species or genus level.

For example, in the Fish (Actinopterygii) group, species like Trachurus japonicus were considered common, yet they occurred sporadically. Meanwhile, in the Gastropoda group, Hermissenda emurai and Clione sp. represented clear findings, with a noted frequency of discovery for each. These statistics indicate significant differences in the biogeographic distribution of taxa within specific environments.

Species Analysis Methods Using DNA Techniques

DNA analysis techniques are modern tools that enhance researchers’ ability to identify species more accurately, particularly in marine ecosystems. These techniques utilize small DNA particles released by living organisms in their environment, allowing for comprehensive information gathering about the present species.

Research has shown that using DNA sequencing techniques in seawater provides a better understanding of biodiversity, and processing species data enhances the ability to discover new species or those that are typically difficult to recognize. In the Ulleung Basin, the analysis of DNA quantities from water samples combined with fishing network results provided a comprehensive picture of biodiversity.

The discovery of 119 species shared between the two methods indicates the importance of combining both the collection and DNA approaches in studies. In similar studies, it was identified that in some categories, such as Copepoda, species were more commonly found in nets, whereas other categories, such as Actinopterygii, were discovered through seawater samples. This highlights their importance in providing new insights into the nature of biodiversity.

Analysis

Biological Diversity According to Data Collection Method

Indicators of biological diversity are important tools for assessing the health status of ecosystems. Using indicators such as the Shannon index and ACE, diversity was measured among the techniques used; results indicated that species diversity was higher in certain months like July, indicating seasonal variation in biodiversity.

Additionally, the statistical analysis of the data using statistical tests helped identify significant differences in diversity based on the sampling method. It was found that these differences reflect the different environmental backgrounds that either reflect the ability to collect species or the diversity of their habitats.

The results obtained show noticeable fluctuations in diversity over time, reflecting the direct impact of environmental factors on the presence of different species. Results from various sampling methods were compared using system validity analysis, helping in a deeper understanding of the changes in species diversity.

The Importance of Continuous Updates to Reference DNA Data

The effectiveness of species identification techniques through DNA sequencing largely depends on the updating of reference databases. These updates are essential to enhance the accuracy and objectivity of the results. Based on data analysis, it shows that the links between DNA sequences and species names must be continuously renewed, which helps to reduce the category of unknown species.

The challenges associated with cryptic species and differences between the available information require ongoing efforts from the research community to update the databases and improve their accuracy. People in this field rely on local and international sources like GenBank and BOLD to enhance taxonomic models and elevate the level of analysis accuracy.

This regular updating effectively contributes to the identification of species that have never been recorded before, meaning that these methods are not only a means of documentation but also for future research. For example, new research adopting modern DNA methods provides new insights into unexpected diversity or endangered species.

Environmental DNA (eDNA) Sampling Techniques

Environmental DNA (eDNA) sampling techniques are considered revolutionary tools in modern ecology, providing a non-invasive way to collect data about living species by analyzing genetic material gathered from the environment. eDNA differs from traditional data collection methods, such as fishing or nets, as DNA is obtained from biological particles released by species into their surroundings, such as dead cells, feces, or bodily secretions. This allows researchers to identify species present in a particular area with high precision, facilitating their study without harming the natural environment. eDNA analysis is particularly effective in the sea where some marine organisms (like pelagic fish) are highly mobile and difficult to collect using traditional nets.

An example of this is a study where eDNA was used to examine marine species around the Korean Peninsula, revealing 350 species of marine animals. This study demonstrated high accuracy in species identification, reaching over 99%. Thus, the use of eDNA can revolutionize how we understand and assess underwater biodiversity.

Comparison Between Bulk DNA and eDNA Techniques

When comparing bulk DNA techniques with eDNA techniques, it becomes evident that each has its strengths and weaknesses. While bulk DNA techniques are useful for collecting large quantities of sample, allowing for a wide range of species to be gathered, this technique may miss some rare or fast-moving species. In contrast, eDNA techniques provide greater access to species that may be difficult to collect using traditional nets. For example, experimental discoveries have indicated that some species, such as jellyfish and other invertebrates, were found in abundance through eDNA analyses compared to bulk DNA analyses.

Results show that…

The research shows that seven groups including Actinopterygii, Cephalopoda, and Scyphozoa exhibited a greater number of species identified through eDNA. This is attributed to the fact that Actinopterygii fish and Cephalopoda mollusks, due to their swimming speed, render traditional net collection methods less effective. However, by collecting water and searching for suspended DNA fragments, they can be accurately identified.

Impact of Sample Conflict on Biodiversity Assessment

The results indicate that variation in the types of samples used can significantly contribute to the assessment of biodiversity. For example, combining eDNA techniques with bulk DNA can provide a broader and more comprehensive view of biodiversity in a specific area. Integrating these diverse methods is considered a necessary step to obtain accurate and comprehensive data on biodiversity.

However, researchers must also be cautious about how to interpret the data collected. The results of the samples should be analyzed carefully, as differences in collection methods can lead to significant variances in final results. For example, in cases where certain species release large amounts of eDNA, these species may appear disproportionately in the results, leading to misleading estimates of biodiversity.

Future Applications of eDNA Techniques

With ongoing advancements in understanding marine environments and ecosystems, there are tremendous possibilities for applying eDNA techniques in studying biodiversity. These techniques can be used in monitoring endangered species or those subjected to dramatic environmental changes, where eDNA can provide valuable information about species presence and mitigate the impacts of overfishing or pollution.

Moreover, eDNA techniques can be a powerful tool in environmental protection, as they allow scientists to identify areas of high biological diversity, helping to direct efforts to protect them. Additionally, these techniques can contribute to understanding climate change and its impact on marine ecosystems, providing deeper insight into how these changes affect species and marine organisms in general.

Current research illustrates the importance of investing more time and resources in these techniques to enrich environmental knowledge and support sustainable policies that contribute to the protection of marine environments and enhance biodiversity. Considering the increasing reliance on eDNA techniques in research, it appears we are on the brink of a new chapter in studying and assessing biodiversity, leading to a better understanding of how to protect these fragile systems.

Marine Ecosystems

Marine ecosystems are vital components supporting life on planet Earth. These systems include oceans, seas, and other water bodies, along with the interactions of the living organisms within them. Marine ecosystems are home to a wide variety of living organisms, ranging from microbes to large mammals like whales. These systems play a crucial role in achieving ecological balance by providing multiple environmental services, such as climate regulation, oxygen production, and water quality improvement.

Marine systems are affected by several natural and human factors, including climate change, pollution, and overfishing. For instance, the increase in ocean temperatures due to global warming leads to coral bleaching, which is a crucial habitat for a wide range of marine organisms. Additionally, overfishing can result in decreasing populations of fish and other marine creatures, negatively impacting marine food webs.

Common examples of marine ecosystems include coral reefs and estuaries. Coral reefs represent the “rainforests” of the oceans, hosting thousands of marine species, and play an essential role in the sustainability of marine life. Meanwhile, estuaries reflect dynamic areas that may contain high biological diversity and economically significant resources such as fisheries.

Growth

Using DNA Technology in Biodiversity Studies

With the rapid advancement of technology, scientists can now use DNA analysis methods to identify marine organism diversity more accurately and easily. Techniques such as “Environmental DNA sequencing” (eDNA) are used to collect data on biodiversity in marine environments without the need to collect living organisms using traditional methods. This technique relies on collecting water samples and analyzing them for traces of DNA from organisms present in that environment.

Many studies indicate that environmental DNA techniques can be more effective than traditional methods in documenting biodiversity, as they allow for quick and accurate data collection about the species present. For example, scientists have used this technique to identify patterns of fish diversity in a particular area, enabling them to obtain precise results within a short timeframe compared to traditional collection methods.

Environmental DNA research helps in managing marine resources by providing accurate data on species numbers and distributions, thus enabling policymakers to make data-driven decisions on how to conserve biodiversity and natural resources. These methods also contribute to conservation efforts, allowing researchers to identify threatened and endangered species more rapidly.

Human Impacts on Marine Biodiversity

Human activities pose a significant threat to marine biodiversity. These activities range from pollution and rising ocean temperatures to overfishing and habitat destruction. One of the notable impacts of pollution is the accumulation of plastic in the oceans, which negatively affects marine life. Chemical pollution, including agricultural chemicals and heavy metals, can impact entire ecosystems, leading to declines in the health of organisms and noticeable effects on marine food chains.

In addition to pollution, unsustainable fishing practices contribute to the decline of several marine species. Overfishing can lead to a sharp decrease in fish populations, which in turn affects other organisms that rely on them for food. Furthermore, habitat destruction due to construction and coastal development leads to the loss of natural marine habitats, increasing pressures on marine organisms.

To mitigate these impacts, environmental policies play a crucial role in managing marine resources. Countries should adopt sustainable strategies to protect marine biodiversity, such as designating protected areas, banning fishing during breeding seasons, and controlling pollution. Many international initiatives, such as agreements stemming from the United Nations Convention on the Law of the Sea, aim to enhance cooperation between countries to protect the oceans and achieve sustainability.

Source link: https://www.frontiersin.org/journals/marine-science/articles/10.3389/fmars.2024.1351148/full

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