no like Aquacultural facilities (example is cage), thier species of fish (Tilapia and bangus), then location (freshwater)

Directions: Research on the different aquaculture facilities and practices commonly undertaken in the country today: Aquaculture Facilities Species of Fish Location Cage Tilapia, Bangus Freshwater or Open salt water

gimme five answers for each 1.Aquacultural facilities, their species of fish, and location

2. Materials and methods2.1. Fish collection and preparationA total of 100 individual fish, 10 per species, were collected from a seashore fish market in Kuakata, Bangladesh, in September 2019. The fish are broadly classified as demersal species (Priacanthus hamrur, Sciades sona and Carangoides chrysophrys), benthopelagic species (Harpadon nehereus and Otolithoides pama), pelagic-neritic species (Setipinna tenuifilis, Coilia neglecta, Anodontostoma chacunda and Sardinella brachysoma) and pelagic species (Megalaspis cordyla) (www.fishbase.org). The fish were caught in the North Bay of Bengal by local fishermen. These species were selected due to their different habitat, availability throughout the year, and economic importance (www.fishbase.org). According to the local fisherman information, people living along the Bay of Bengal’s coastline are routinely consuming the fish species throughout the years. Information on the fish species is shown in Table 1. The collected fish were wrapped with aluminum foil, placed in an icebox, and transported to the laboratory of the Department of Environmental Science and Technology, Jashore University of Science and Technology, and then dissected or frozen (–20 °C) and later thawed. In the laboratory, each fish sample was cleaned in MPs free distilled water to remove any externally attached plastic as recommended by Lusher et al. (2016). The body length (cm) and weight (g) of each fish were recorded. Each fish was then opened on a wood tray, using scissors, scalpel and forceps, and the gastrointestinal tract (GIT) was removed, and then transferred to a Petri dish and weighed (g). Finally, the GIT was placed in 500 mL glass beaker for MPs extraction and covered with aluminum foil to minimize the risk of contamination.Table 1. Information on studied fish species and their corresponding levels of microplastics ingestion.Species name Sample Habitat Mean length Length range Mean weight Weight range Mean GIT weight Total Microplastics (MPs) Mean MPs /individual Mean MPs/gBW Mean MPs/gGITEmpty Cell (n) Empty Cell (cm) ± SD (cm) (g) ± SD (g) (g) ± SD Empty Cell Empty Cell Empty Cell Empty CellPriacanthus hamrur 10 Demersal 15.8 ± 0.7 14.7–16.6 41.0 ± 10.6 27.4–65.1 1.5 ± 0.4 38 3.8 0.09 2.53Setipinna tenuifilis 10 Pelagic-neritic 14.0 ± 1.0 13.5–16.6 22.0 ± 4.0 16.5–27.8 0.5 ± 0.2 32 3.2 0.15 6.45Sciades sona 10 Demersal 18.7 ± 0.7 17.8–19.6 28.9 ± 6.3 18.1–38.2 1.8 ± 0.3 30 3.0 0.10 1.67Carangoides chrysophrys 10 Demersal 12.5 ± 1.0 11.4–14.0 23.7 ± 4.0 17.2–28.3 0.8 ± 0.1 20 2.0 0.08 2.50Sardinella brachysoma 10 Pelagic-neritic 15.0 ± 0.5 14.1–15.7 35.0 ± 2.9 31.1–40.8 1.1 ± 0.2 20 2.0 0.06 1.82Harpadon nehereus 10 Benthopelagic 22.2 ± 1.8 19.1–24.6 58.0 ± 35.4 10.7–103.3 1.8 ± 0.8 18 1.8 0.03 1.00Otolithoides pama 10 Benthopelagic 18.8 ± 1.0 17.6–20.1 33.7 ± 6.5 24.2–44.0 1.5 ± 0.3 18 1.8 0.05 1.20Coilia neglecta 10 Pelagic-neritic 19.5 ± 1.4 17.5–21.4 38.5 ± 3.2 35.0–44.1 1.6 ± 0.4 15 1.5 0.04 0.94Anodontostoma chacunda 10 Pelagic-neritic 15.5 ± 1.0 14.2–17.1 55.5 ± 5.6 44.9–66.9 3.1 ± 0.4 14 1.4 0.03 0.45Megalaspis cordyla 10 Pelagic 18.2 ± 1.2 16.5–19.6 55.2 ± 9.8 39.85–66.0 1.6 ± 0.3 10 1.0 0.02 0.632.2. Microplastics extractionMPs from the fish GITs were extracted using the peroxide oxidation-based digestion method described in several studies (eg. Avio et al., 2015, McNeish et al., 2018, Nuelle et al., 2014) with minor modifications because of its superiority over acid (eg. HCl) and alkali (eg. NaOH) digestion, and minimal impact on MPs (Cole et al., 2014). In brief, firstly, the GITs were dried at 75 °C for 24 h in an oven. After that, the wet peroxide oxidation process was employed using 50 mL of 0.05 M Fe (II) and 50 mL of 30% hydrogen peroxide (H2O2) at 65 °C on a hotplate to remove the organic matter through digestion. In case, if there remains some visible organic matter, an additional 10 mL of H2O2 was added. After digestion, the liquid was then passed through a stack of fractionating sieves (mesh sizes: 5000μm, 2000μm, 1000μm, and 500μm) and the filtrate was collected. The residue on the sieve was backwashed into a clean 500 mL glass beaker using MPs free distilled water and metal spatula (Kolandhasamy et al., 2018). The resulting liquid from the sieve backwash and filtrate were then separately passed through glass microfiber filter (GF/B: 1.0μm pore size; 47 mm in diameter; Whatman, UK) using a vacuum filtration unit. All the filter papers were placed into separate glass Petri dishes, closed, covered with aluminum foil papers, and air-dried for further analysis. To minimize potential contamination with MPs, all the washing solutions were filtered through the glass microfiber filter.2.3. Quantification and characterization of microplasticsMPs were identified visually under a binocular microscope (HumaScope Advanced LED, Human, Germany). Each of the particles was counted adjusting the magnification from 10 x to 100 x. The number, color, shape, and size of MPs in each fish species was recorded. The abundance of MPs was calculated as the number of particles per fish species, individuals, body weight and GITs along with the statistical analysis of variances (Table 1).The MPs polymer types of all the extracted particles were identified by Fourier Transform Infrared Spectroscopy (FTIR) (FTIR-4600, JASCO Inc., Tokyo, Japan) equipped with Attenuated Total Reflection (ATR) unit. For the identification of each MP, the spectrum range was from 4000 to 550 cm −1 with a spectral resolution of 4 cm −1. Sixteen co-scans were performed in each measurement. Background cleaning was completed before each particle measurement. Data collection was performed using the SpectraManager ™ II software (JASCO Inc., Tokyo, Japan). Obtained spectra were analyzed by comparing them to polymer spectra from commercial libraries using the KnowItAll® Informatics System 2013 (JASCO Edition, Bio-Rad Laboratories, Inc., Hercules, CA, USA). Each particle was analyzed at least one to three times and results were selected depending on the accuracy of the spectra comparing to the reference library. However, it was difficult to confirm the polymer types by the FTIR of those extracted MPs (n = 11) < 150μm and fibers with a small width (n = 20). Instead, a hot point test proposed by Devriese et al. (2015) was applied following Baalkhuyur et al. (2018) on a sub-sample of the unidentifiable MPs to determine the polymer types.The surface morphology of the identified MPs was observed using a field emission scanning electron microscope (FE-SEM, Zeiss Sigma, Carl Zeiss, Germany). Since examination of all the extracted MPs by FE-SEM is expensive and time consuming, a total of 60 MPs from the typical representatives of different colors, sizes, and shapes were chosen randomly upon microscopic observation. The selected MPs were coated with a thin film of evaporated gold using an ion sputtering instrument (Hitachi E-1045, Japan) and viewed with an accelerating voltage up to 10.00 kV.Download : Download high-res image (187KB)Download : Download full-size imageFig. 1. Bivariate plots of extracted microplastics per fish species against (A) per unit gastrointestinal tract (GIT) weight, and (B) per unit body weight (BW). Solid line depicts the predicted relationship; and shaded regions are the 95% confidence intervals (total 10 species, and 10 individual per species were investigated).Download : Download high-res image (824KB)Download : Download full-size imageFig. 2. The distribution (%) of microplastics extracted from the gastrointestinal tracts of ten fish species: (A) shape, (B) color and (C) size. The numbers in brackets correspond to the pieces of plastic found for each species.2.4. Quality assurance and quality controlFish dissection and separation of GIT were done inside a clean bench and the GIT was placed in a Petri dish and covered to prevent contamination. Cotton cloths, latex gloves, glassware, and metalware were used at the laboratory. All the liquids used in the experiments were filtered through GF/B (1.0μm pore size; 47 mm in diameter) glass microfiber filter (Whatman, UK) before use. All the glassware, dissection tools, and filtration units were rinsed three times with filtered water. One blank sample without fish GIT was prepared following the same procedure to correct the potential procedural contamination, if any. One of the limitation of this study is that all the fish were collected from fishermen in an open fish market and there were no controls over MPs contamination from the time of being harvested to arrival to the market. As a result, each fish sample was cleaned in MPs free distilled water to remove any externally attached plastic.2.5. Statistical analysisThe abundance and characteristics data of MPs were recorded for each fish species (Table 1). The average number of MPs per 1 g wet weight of body (BW) and GIT were also calculated for each species (Table 1). Before statistical analysis, the assumption of homogeneity of variances was tested using Shapiro–Wilk test and the data conformed to a normal distribution (Shapiro–Wilk, p > 0.05). The fish samples were classified based on their species, which was then used as a fixed factor in factorial Analyses of Variance (ANOVA). Differences between the means were considered to be statistically significant when p < 0.05. Bivariate plots of mean MPs abundance in the fish species against per unit GIT weight, and per unit BW were analyzed at the 95% confidence intervals (Pearson, p < 0.05).Statistical analysis was performed using R Studio (with R version 3.5.1, www.rstudio.com), and figures were generated using ggplot2 (version 3.1.0).

All the following are aquatic ecosystem exceptQuestion 5Answera.Fresh waterb.Estuarinec.Plateaud.Marine

Where does the majority of aquaculture occur?Coastal waters.Deep ocean watersFreshwater ecosystems.Brackish waters.