Search results for: recirculating aquaculture
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 190

Search results for: recirculating aquaculture

10 Wildlife Communities in the Service of Extensively Managed Fishpond Systems – Advantages of a Symbiotic Relationship

Authors: Peter Palasti, Eva Kerepeczki

Abstract:

Extensive fish farming is one of the most traditional forms of aquaculture in Europe, usually practiced in large pond systems with earthen beds, where the growth of fish is based on natural feed and supplementary foraging. These farms have semi-natural environmental conditions, sustaining diverse wildlife communities that have complex effects on fish production and also provide a livelihood for many wetland related taxa. Based on their characteristics, these communities could be sources of various ecosystem services (ESs), that could also enhance the value and enable the multifunctional use of these artificially constructed and maintained production zones. To identify and estimate the whole range of wildlife’s contribution we have conducted an integrated assessment in an extensively managed pond system in Biharugra, Hungary, where we studied 14 previously revealed ESs: fish and reed production, water storage, water and air quality regulation, CO2 absorption, groundwater recharge, aesthetics, recreational activities, inspiration, education, scientific research, presence of semi-natural habitats and useful/protected species. ESs were collected through structured interviews with the local experts of all major stakeholder groups, where we have also gathered information about the known forms, levels (none, low, high) and orientations (positive, negative) of the contributions of the wildlife community. After that, a quantitative analysis was carried out: we calculated the total mean value of the services being used between 2014-16, then we estimated the value and percentage of contributions. For the quantification, we mainly used biophysical indicators with the available data and empirical knowledge of the local experts. During the interviews, 12 of the previously listed services (85%) were mentioned to be related to wildlife community, consisting of 5 fully (e.g., recreation, reed production) and seven partially dependent ESs (e.g., inspiration, CO2 absorption) from our list. The orientation of the contributions was said to be positive almost every time; however, in the case of fish production, the feeding habit of some wild species (Phalacrocorax carbo, Lutra lutra) caused significant losses in fish stocks in the study period. During the biophysical assessment, we calculated the total mean value of the services and quantified the aid of wildlife community at the following services: fish and reed production, recreation, CO2 absorption, and the presence of semi-natural habitats and wild species. The combined results of our interviews and biophysical evaluations showed that the presence of wildlife community not just greatly increased the productivity of the fish farms in Biharugra (with ~53% of natural yield generated by planktonic and benthic communities) but also enhanced the multifunctionality of the system through expanding the quality and number of its services. With these abilities, extensively managed fishponds could play an important role in the future as refugia for wetland related services and species threatened by the effects of global warming.

Keywords: ecosystem services, fishpond systems, integrated assessment, wildlife community

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9 Analysis of Taxonomic Compositions, Metabolic Pathways and Antibiotic Resistance Genes in Fish Gut Microbiome by Shotgun Metagenomics

Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

Abstract:

Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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8 Glucose Measurement in Response to Environmental and Physiological Challenges: Towards a Non-Invasive Approach to Study Stress in Fishes

Authors: Tomas Makaras, Julija Razumienė, Vidutė Gurevičienė, Gintarė Sauliutė, Milda Stankevičiūtė

Abstract:

Stress responses represent animal’s natural reactions to various challenging conditions and could be used as a welfare indicator. Regardless of the wide use of glucose measurements in stress evaluation, there are some inconsistencies in its acceptance as a stress marker, especially when it comes to comparison with non-invasive cortisol measurements in the fish challenging stress. To meet the challenge and to test the reliability and applicability of glucose measurement in practice, in this study, different environmental/anthropogenic exposure scenarios were simulated to provoke chemical-induced stress in fish (14-days exposure to landfill leachate) followed by a 14-days stress recovery period and under the cumulative effect of leachate fish subsequently exposed to pathogenic oomycetes (Saprolegnia parasitica) to represent a possible infection in fish. It is endemic to all freshwater habitats worldwide and is partly responsible for the decline of natural freshwater fish populations. Brown trout (Salmo trutta fario) and sea trout (Salmo trutta trutta) juveniles were chosen because of a large amount of literature on physiological stress responses in these species was known. Glucose content in fish by applying invasive and non-invasive glucose measurement procedures in different test mediums such as fish blood, gill tissues and fish-holding water were analysed. The results indicated that the quantity of glucose released in the holding water of stressed fish increased considerably (approx. 3.5- to 8-fold) and remained substantially higher (approx. 2- to 4-fold) throughout the stress recovery period than the control level suggesting that fish did not recover from chemical-induced stress. The circulating levels of glucose in blood and gills decreased over time in fish exposed to different stressors. However, the gill glucose level in fish showed a decrease similar to the control levels measured at the same time points, which was found to be insignificant. The data analysis showed that concentrations of β-D glucose measured in gills of fish treated with S. parasitica differed significantly from the control recovery, but did not differ from the leachate recovery group showing that S. parasitica presence in water had no additive effects. In contrast, a positive correlation between blood and gills glucose were determined. Parallel trends in blood and water glucose changes suggest that water glucose measurement has much potency in predicting stress. This study demonstrated that measuring β-D-glucose in fish-holding water is not stressful as it involves no handling and manipulation of an organism and has critical technical advantages concerning current (invasive) methods, mainly using blood samples or specific tissues. The quantification of glucose could be essential for studies examining the stress physiology/aquaculture studies interested in the assessment or long-term monitoring of fish health.

Keywords: brown trout, landfill leachate, sea trout, pathogenic oomycetes, β-D-glucose

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7 Defense Priming from Egg to Larvae in Litopenaeus vannamei with Non-Pathogenic and Pathogenic Bacteria Strains

Authors: Angelica Alvarez-Lee, Sergio Martinez-Diaz, Jose Luis Garcia-Corona, Humberto Lanz-Mendoza

Abstract:

World aquaculture is always looking for improvements to achieve productions with high yields avoiding the infection by pathogenic agents. The best way to achieve this is to know the biological model to create alternative treatments that could be applied in the hatcheries, which results in greater economic gains and improvements in human public health. In the last decade, immunomodulation in shrimp culture with probiotics, organic acids and different carbon sources has gained great interest, mainly in larval and juvenile stages. Immune priming is associated with a strong protective effect against a later pathogen challenge. This work provides another perspective about immunostimulation from spawning until hatching. The stimulation happens during development embryos and generates resistance to infection by pathogenic bacteria. Massive spawnings of white shrimp L. vannamei were obtained and placed in experimental units with 700 mL of sterile seawater at 30 °C, salinity of 28 ppm and continuous aeration at a density of 8 embryos.mL⁻¹. The immunostimulating effect of three death strains of non-pathogenic bacterial (Escherichia coli, Staphylococcus aureus and Bacillus subtilis) and a pathogenic strain for white shrimp (Vibrio parahaemolyticus) was evaluated. The strains killed by heat were adjusted to O.D. 0.5, at A 600 nm, and directly added to the seawater of each unit at a ratio of 1/100 (v/v). A control group of embryos without inoculum of dead bacteria was kept under the same physicochemical conditions as the rest of the treatments throughout the experiment and used as reference. The duration of the stimulus was 12 hours, then, the larvae that hatched were collected, counted and transferred to a new experimental unit (same physicochemical conditions but at a salinity of 28 ppm) to carry out a challenge of infection against the pathogen V. parahaemolyticus, adding directly to seawater an amount 1/100 (v/v) of the live strain adjusted to an OD 0.5; at A 600 nm. Subsequently, 24 hrs after infection, nauplii survival was evaluated. The results of this work shows that, after 24 hrs, the hatching rates of immunostimulated shrimp embryos with the dead strains of B. subtillis and V. parahaemolyticus are significantly higher compared to the rest of the treatments and the control. Furthermore, survival of L. vanammei after a challenge of infection of 24 hrs against the live strain of V. parahaemolyticus is greater (P < 0.05) in the larvae immunostimulated during the embryonic development with the dead strains B. subtillis and V. parahaemolyticus, followed by those that were treated with E. coli. In summary superficial antigens can stimulate the development cells to promote hatching and can have normal development in agreeing with the optical observations, plus exist a differential response effect between each treatment post-infection. This research provides evidence of the immunostimulant effect of death pathogenic and non-pathogenic bacterial strains in the rate of hatching and oversight of shrimp L. vannamei during embryonic and larval development. This research continues evaluating the effect of these death strains on the expression of genes related to the defense priming in larvae of L. vannamei that come from massive spawning in hatcheries before and after the infection challenge against V. parahaemolyticus.

Keywords: immunostimulation, L. vannamei, hatching, survival

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6 Effect of Rapeseed Press Cake on Extrusion System Parameters and Physical Pellet Quality of Fish Feed

Authors: Anna Martin, Raffael Osen

Abstract:

The demand for fish from aquaculture is constantly growing. Concurrently, due to a shortage of fishmeal caused by extensive overfishing, fishmeal substitution by plant proteins is getting increasingly important for the production of sustainable aquafeed. Several research studies evaluated the impact of plant protein meals, concentrates or isolates on fish health and fish feed quality. However, these protein raw materials often require elaborate and expensive manufacturing and their availability is limited. Rapeseed press cake (RPC) – a side product of de-oiling processes – exhibits a high potential as a plant-based fishmeal alternative in fish feed for carnivorous species due to its availability, low costs and protein content. In order to produce aquafeed with RPC, it is important to systematically assess i) inclusion levels of RPC with similar pellet qualities compared to fishmeal containing formulations and ii) how extrusion parameters can be adjusted to achieve targeted pellet qualities. However, the effect of RPC on extrusion system parameters and pellet quality has only scarcely been investigated. Therefore, the aim of this study was to evaluate the impact of feed formulation, extruder barrel temperature (90, 100, 110 °C) and screw speed (200, 300, 400 rpm) on extrusion system parameters and the physical properties of fish feed pellets. A co-rotating pilot-scale twin screw extruder was used to produce five iso-nitrogenous feed formulations: a fish meal based reference formulation including 16 g/100g fishmeal and four formulations in which fishmeal was substituted by RPC to 25, 50, 75 or 100 %. Extrusion system parameters, being product temperature, pressure at the die, specific mechanical energy (SME) and torque, were monitored while samples were taken. After drying, pellets were analyzed regarding to optical appearance, sectional and longitudinal expansion, sinking velocity, bulk density, water stability, durability and specific hardness. In our study, the addition of minor amounts of RPC already had high impact on pellet quality parameters, especially on expansion but only marginally affected extrusion system parameters. Increasing amounts of RPC reduced sectional expansion, sinking velocity, bulk density and specific hardness and increased longitudinal expansion compared to a reference formulation without RPC. Water stability and durability were almost not affected by RPC addition. Moreover, pellets with rapeseed components showed a more coarse structure than pellets containing only fishmeal. When the adjustment of barrel temperature and screw speed was investigated, it could be seen that the increase of extruder barrel temperature led to a slight decrease of SME and die pressure and an increased sectional expansion of the reference pellets but did almost not affect rapeseed containing fish feed pellets. Also changes in screw speed had little effects on the physical properties of pellets however with raised screw speed the SME and the product temperature increased. In summary, a one-to-one substitution of fishmeal with RPC without the adjustment of extrusion process parameters does not result in fish feed of a designated quality. Therefore, a deeper knowledge of raw materials and their behavior under thermal and mechanical stresses as applied during extrusion is required.

Keywords: extrusion, fish feed, press cake, rapeseed

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5 Bacteriophages for Sustainable Wastewater Treatment: Application in Black Water Decontamination with an Emphasis to DRDO Biotoilet

Authors: Sonika Sharma, Mohan G. Vairale, Sibnarayan Datta, Soumya Chatterjee, Dharmendra Dubey, Rajesh Prasad, Raghvendra Budhauliya, Bidisha Das, Vijay Veer

Abstract:

Bacteriophages are viruses that parasitize specific bacteria and multiply in metabolising host bacteria. Bacteriophages hunt for a single or a subset of bacterial species, making them potential antibacterial agents. Utilizing the ability of phages to control bacterial populations has several applications from medical to the fields of agriculture, aquaculture and the food industry. However, harnessing phage based techniques in wastewater treatments to improve quality of effluent and sludge release into the environment is a potential area for R&D application. Phage mediated bactericidal effect in any wastewater treatment process has many controlling factors that lead to treatment performance. In laboratory conditions, titer of bacteriophages (coliphages) isolated from effluent water of a specially designed anaerobic digester of human night soil (DRDO Biotoilet) was successfully increased with a modified protocol of the classical double layer agar technique. Enrichment of the same was carried out and efficacy of the phage enriched medium was evaluated at different conditions (specific media, temperature, storage conditions). Growth optimization study was carried out on different media like soybean casein digest medium (Tryptone soya medium), Luria-Bertani medium, phage deca broth medium and MNA medium (Modified nutrient medium). Further, temperature-phage yield relationship was also observed at three different temperatures 27˚C, 37˚C and 44˚C at laboratory condition. Results showed the higher activity of coliphage 27˚C and at 37˚C. Further, addition of divalent ions (10mM MgCl2, 5mM CaCl2) and 5% glycerol resulted in a significant increase in phage titer. Besides this, effect of antibiotics addition like ampicillin and kanamycin at different concentration on plaque formation was analysed and reported that ampicillin at a concentration of 1mg/ml ampicillin stimulates phage infection and results in more number of plaques. Experiments to test viability of phage showed that it can remain active for 6 months at 4˚C in fresh tryptone soya broth supplemented with fresh culture of coliforms (early log phase). The application of bacteriophages (especially coliphages) for treatment of effluent of human faecal matter contaminated effluent water is unique. This environment-friendly treatment system not only reduces the pathogenic coliforms, but also decreases the competition between nuisance bacteria and functionally important microbial populations. Therefore, the phage based cocktail to treat fecal pathogenic bacteria present in black water has many implication in wastewater treatment processes including ‘DRDO Biotoilet’, which is an ecofriendly appropriate and affordable human faecal matter treatment technology for different climates and situations.

Keywords: wastewater, microbes, virus, biotoilet, phage viability

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4 Ecological Planning Method of Reclamation Area Based on Ecological Management of Spartina Alterniflora: A Case Study of Xihu Harbor in Xiangshan County

Authors: Dong Yue, Hua Chen

Abstract:

The study region Xihu Harbor in Xiangshan County, Ningbo City is located in the central coast of Zhejiang Province. Concerning the wave dispating issue, Ningbo government firstly introduced Spartina alterniflora in 1980s. In the 1990s, S. alterniflora spread so rapidly thus a ‘grassland’ in the sea has been created nowadays. It has become the most important invasive plant of China’s coastal tidal flats. Although S. alterniflora had some ecological and economic functions, it has also brought series of hazards. It has ecological hazards on many aspects, including biomass and biodiversity, hydrodynamic force and sedimentation process, nutrient cycling of tidal flat, succession sequence of soil and plants and so on. On engineering, it courses problems of poor drainage and channel blocking. On economy, the hazard mainly reflected in the threat on aquaculture industry. The purpose of this study is to explore an ecological, feasible and economical way to manage Spartina alterniflora and use the land formed by it, taking Xihu Harbor in Xiangshan County as a case. Comparison method, mathematical modeling, qualitative and quantitative analysis are utilized to proceed the study. Main outcomes are as follows. By comparing a series of S. alterniflora managing methods which include the combination of mechanical cutting and hydraulic reclamation, waterlogging, herbicide and biological substitution from three standpoints – ecology, engineering and economy. It is inferred that the combination of mechanical cutting and hydraulic reclamation is among the top rank of S. alternifora managing methods. The combination of mechanical cutting and hydraulic reclamation means using large-scale mechanical equipment like large screw seagoing dredger to excavate the S. alterniflora with root and mud together. Then the mix of mud and grass was blown off nearby coastal tidal zone transported by pipelines, which can cushion the silt of tidal zone to form a land. However, as man-made land by coast, the reclamation area’s ecological sensitivity is quite high and will face high possibility of flood threat. Therefore, the reclamation area has many reasonability requirements, including ones on location, specific scope, water surface rate, direction of main watercourse, site of water-gate, the ratio of ecological land to urban construction land. These requirements all became important basis when the planning was being made. The water system planning, green space system planning, road structure and land use all need to accommodate the ecological requests. Besides, the profits from the formed land is the managing project’s source of funding, so how to utilize land efficiently is another considered point in the planning. It is concluded that by aiming at managing a large area of S. alterniflora, the combination of mechanical cutting and hydraulic reclamation is an ecological, feasible and economical method. The planning of reclamation area should fully respect the natural environment and possible disasters. Then the planning which makes land use efficient, reasonable, ecological will promote the development of the area’s city construction.

Keywords: ecological management, ecological planning method, reclamation area, Spartina alternifora, Xihu harbor

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3 The Impact of β Nucleating Agents and Carbon-Based Nanomaterials on Water Vapor Permeability of Polypropylene Composite Films

Authors: Glykeria A. Visvini, George Ν. Mathioudakis, Amaia Soto Beobide, George A. Voyiatzis

Abstract:

Polymer nanocomposites are materials in which a polymer matrix is reinforced with nanoscale inclusions, such as nanoparticles, nanoplates, or nanofibers. These nanoscale inclusions can significantly enhance the mechanical, thermal, electrical, and other properties of the polymer matrix, making them attractive for a wide range of industrial applications. These properties can be tailored by adjusting the type and the concentration of the nanoinclusions, which provides a high degree of flexibility in their design and development. An important property that polymeric membranes can exhibit is water vapor permeability (WVP). This can be accomplished by various methods, including the incorporation of micro/nano-fillers into the polymer matrix. In this way, a micro/nano-pore network can be formed, allowing water vapor to permeate through the membrane. At the same time, the membrane can be stretched uni- or bi-axially, creating aligned or cross-linked micropores in the composite, respectively, which can also increase the WVP. Nowadays, in industry, stretched films reinforced with CaCO3 develop micro-porosity sufficient to give them breathability characteristics. Carbon-based nanomaterials, such as graphene oxide (GO), are tentatively expected to be able to effectively improve the WVP of corresponding composite polymer films. The presence in the GO structure of various functional oxidizing groups enhances its ability to attract and channel water molecules, exploiting the unique large surface area of graphene that allows the rapid transport of water molecules. Polypropylene (PP) is widely used in various industrial applications due to its desirable properties, including good chemical resistance, excellent thermal stability, low cost, and easy processability. The specific properties of PP are highly influenced by its crystalline behavior, which is determined by its processing conditions. The development of the β-crystalline phase in PP, in combination with stretching, is anticipating improving the microporosity of the polymer matrix, thereby enhancing its WVP. The aim of present study is to create breathable PP composite membranes using carbon-based nanomaterials, such as graphene oxide (GO), reduced graphene oxide (rGO), and graphene nanoplatelets (GNPs). Unlike traditional methods that rely on the drawing process to enhance the WVP of PP, this study intents to develop a low-cost approach using melt mixing with β-nucleating agents and carbon fillers to create highly breathable PP composite membranes. The study aims to investigate how the concentration of these additives affects the water vapor transport properties of the resulting PP films/membranes. The presence of β-nucleating agents and carbon fillers is expected to enhance β-phase growth in PP, while an alternation between β- and α-phase is expected to lead to improved microporosity and WVP. Our ambition is to develop highly breathable PP composite films with superior performance and at a lower cost compared to the benchmark. Acknowledgment: This research has been co‐financed by the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call «Special Actions "AQUACULTURE"-"INDUSTRIAL MATERIALS"-"OPEN INNOVATION IN CULTURE"» (project code: Τ6YBP-00337)

Keywords: carbon based nanomaterials, nanocomposites, nucleating agent, polypropylene, water vapor permeability

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2 Immunostimulatory Response of Supplement Feed in Fish against Aeromonas hydrophila

Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

Abstract:

Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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1 Development of Anti-Fouling Surface Features Bioinspired by the Patterned Micro-Textures of the Scophthalmus rhombus (Brill)

Authors: Ivan Maguire, Alan Barrett, Alex Forte, Sandra Kwiatkowska, Rohit Mishra, Jens Ducrèe, Fiona Regan

Abstract:

Biofouling is defined as the gradual accumulation of Biomimetics refers to the use and imitation of principles copied from nature. Biomimetics has found interest across many commercial disciplines. Among many biological objects and their functions, aquatic animals deserve a special attention due to their antimicrobial capabilities resulting from chemical composition, surface topography or other behavioural defences, which can be used as an inspiration for antifouling technology. Marine biofouling has detrimental effects on seagoing vessels, both commercial and leisure, as well as on oceanographic sensors, offshore drilling rigs, and aquaculture installations. Sensor optics, membranes, housings and platforms can become fouled leading to problems with sensor performance and data integrity. While many anti-fouling solutions are currently being investigated as a cost-cutting measure, biofouling settlement may also be prevented by creating a surface that does not satisfy the settlement conditions. Brill (Scophthalmus rhombus) is a small flatfish occurring in marine waters of Mediterranean as well as Norway and Iceland. It inhabits sandy and muddy coastal waters from 5 to 80 meters. Its skin colour changes depending on environment, but generally is brownish with light and dark freckles, with creamy underside. Brill is oval in shape and its flesh is white. The aim of this study is to translate the unique micro-topography of the brill scale, to design marine inspired biomimetic surface coating and test it against a typical fouling organism. Following extensive study of scale topography of the brill fish (Scophthalmus rhombus) and the settlement behaviour of the diatom species Psammodictyon sp. via SEM, two state-of-the-art antifouling surface solutions were designed and investigated; A brill fish scale bioinspired surface pattern platform (BFD), and generic and uniformly-arrayed, circular micropillar platform (MPD), with offsets based on diatom species settlement behaviour. The BFD approach consists of different ~5 μm by ~90 μm Brill-replica patterns, grown to a 5 μm height, in a linear array pattern. The MPD approach utilises hexagonal-packed cylindrical pillars 10.6 μm in diameter, grown to a height of 5 μm, with vertical offset of 15 μm and horizontal offset of 26.6 μm. Photolithography was employed for microstructure growth, with a polydimethylsiloxane (PDMS) chip-based used as a testbed for diatom adhesion on both platforms. Settlement and adhesion tests were performed using this PDMS microfluidic chip through subjugation to centrifugal force via an in-house developed ‘spin-stand’ which features a motor, in combination with a high-resolution camera, for real-time observing diatom release from PDMS material. Diatom adhesion strength can therefore be determined based on the centrifugal force generated at varying rotational speeds. It is hoped that both the replica and bio-inspired solutions will give comparable anti-fouling results to these synthetic surfaces, whilst also assisting in determining whether anti-fouling solutions should predominantly be investigating either fully bioreplica-based, or a bioinspired, synthetically-based design.

Keywords: anti-fouling applications, bio-inspired microstructures, centrifugal microfluidics, surface modification

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