A synbiotic containing prebiotic prepared from a by-product of king oyster mushroom, Pleurotus eryngii and probiotic, Lactobacillus plantarum incorporated in diet to improve the growth performance and health status of white shrimp, Litopenaeus vannamei.

This study aimed to evaluate the effects of a synbiotic composite an extract from a by-product of king oyster mushroom, Pleurotus eryngii (KOME), and probiotic Lactobacillus plantarum 7-40 on the growth performance and health status of white shrimp, Litopenaeus vannamei. The KOME was able to stimulate the growth of probiotic, but not the growth of Vibrio pathogens, including V. alginolyticus, V. parahaemolyticus, and V. harveyi. Four diets were formulated, including a control diet supplemented without prebiotic and probiotic, a basal diet supplemented with KOME (5 g kg-1) (ME), a basal diet supplemented with probiotic (1 × 108 CFU kg-1) (LP), and a basal diet supplemented with KOME (5 g kg-1) and probiotic (1 × 108 CFU kg-1) (SYN). Shrimp fed the ME, LP, and SYN diets had significantly higher survival than that of shrimp fed with the control diet for 8 weeks. Shrimp in the SYN group also had a significantly higher weight gain and total final weight in comparison with the control and other treatments. In the intestinal tract, lactic acid bacteria count was significantly higher in the SYN group, whereas the Vibrio-like bacteria count was significantly higher in the ME group than in the control group. For the health status assessment, the disease resistance of shrimp against V. alginolyticus was improved in all treatments compared to the shrimp in control. Shrimps in the SYN group had significantly lower cumulative mortality due to the significant increase in immune responses, including phenoloxidase, respiratory burst, and lysozyme activity, and the gene expression of pexn and pen4 in the haemocytes, and lgbp, sp, propoii, pexn, pen3a, pen4, and gpx in the haepatopancreas of shrimp as compared to the control. Therefore, it is suggested that a combination of KOME and probiotics can be used as a synbiotic to improve the growth performance and reduce the risk of infectious diseases caused by Vibrio and at the same time significantly contribute to the circular economy.

Exopolysaccharide from Porphyridium cruentum (purpureum) is Not Toxic and Stimulates Immune Response against Vibriosis: The Assessment Using Zebrafish and White Shrimp Litopenaeus vannamei.

Exopolysaccharides, or extracellular polysaccharides (EPS, sPS), represent a valuable metabolite compound synthesized from red microalgae. It is a non-toxic natural agent and can be applied as an immunostimulant. The toxicity test of exopolysaccharides from Porphyridium has been done in vivo using zebrafish (Danio rerio) embryonic model, or the ZET (zebrafish embryotoxicity test). The administration of extracellular polysaccharides or exopolysaccharides (EPS) from microalgae Porphyridium cruentum (synonym: P. purpureum) to shrimps Litopenaeus vannamei was investigated to determine the effect of this immunostimulant on their non-specific immune response and to test if this compound can be used as a protective agent for shrimps in relation to Vibrio infection. For immune response, exopolysaccharides were given to shrimps via the immersion method on day 1 and booster on day 8. Shrimp hemocytes were taken on day 1 (EPS administration), day 7 (no treatment), day 8 (EPS booster) and day 9 (Vibrio infection) and tested for their immune response on each treatment. The result shows that the EPS is not toxic, as represented by the normal embryonic development and the mortality data. In the Pacific white shrimps, an increase in the values of all immune parameters was shown, in line with the increasing EPS concentration, except for the differential hemocyte count (DHC). In detail, an increase was noted in total hemocytes (THC) value, phagocytotic activity (PA) and respiratory burst (RB) in line with the EPS concentration increase. These results and other previous studies indicate that EPS from Porphyridium is safe, enhances immune parameters in shrimp rapidly, and has the ability to act as an immunostimulant or an immunomodulator. It is a good modulator for the non-specific immune cells of Pacific white shrimps, and it can be used as a preventive agent against vibriosis.

Fucoidan extract from brown seaweed (Sargassum echinocarpum): molecular weight, elemental composition, selectivity and anticancer activity against breast cancer cells.

Molecular weight and elemental composition contribute to fucoidan bioactivity. Fucoidan extract from S. echinocarpum had three fractions varying in molecular weights (Mw), whereas crude fucoidan extract had a moderate Mw of 2,034.31 kDa. The fucoidan extract contained several elements, including C (38.19%), O (44.46%), and S (2.61%). Moreover, the fucoidan extract exhibited toxicity to breast cancer cells (MCF-7) at 297.58 ± 2.40 ppm. The extract induced apoptosis by 49.78%, 72.05%, and 89.35% after incubation for 24, 48, and 72 h, respectively. Fucoidan increased MDA levels in MCF-7 cells, indicating its anticancer properties through the induction of apoptosis-induced lipid peroxidation. Furthermore, the anticancer activity of fucoidan extract was not significantly different from the standard fucoidan (F. vesiculosus). In addition, the extract was selective and non-toxic to human normal cells (TIG 1-20), indicating its safety for consumption and potential as an anticancer agent derived from marine algae.

Characterization of 'novel fucoidan' extracted from brown seaweed (Sargassum echinocarpum J. Ag.) using Ultrasound-Assisted Extraction (UAE) and its potential antioxidant activity.

Fucoidan of Sargassum echinocarpum was extracted using the UAE method. This method produced a fucoidan extract with a yield of 2.82-3.93% and antioxidant activity of 44.57-98.77 ppm at different temperatures and extraction times. The total sulphate, fucose, phenolic, protein, and uronic acid contents were 5.50-6.24%, 17.22-63.22%, 14.66-78.20 mgGAE/g, 0-14.42 mg/g, and 191.43-353.76 mg/g, respectively. Furthermore, extraction treatment with UAE also affected the topology and morphology of the fucoidan extract. The analysis results showed the presence of monosaccharides such as galactose 1.37%, mannose 0.642%, and xylose 2.86%, while glucose and rhamnose were not detected. The extract also had the same absorption band as the fucose and fucoidan standard (F. vesiculosus) as well as a similar basic skeletal structure. Based on the experimental results, the fucoidan extract had excellent antioxidant properties.

In silico molecular docking, molecular dynamics, ADMET analysis of fucoidan against receptor frizzled-8 and coreceptor LRP6 in Wnt/ß-Catenin pathway and in vitro analysis of fucoidan extract from Sargassum echinocarpum as ß-catenin inhibitor in breast cancer cell line (MCF-7).

This study aimed to investigate the effect of fucoidan on the Wnt/ß-Catenin pathway using both in-silico molecular docking, molecular dynamics, ADMET analysis (in frizzled-8 receptor and LRP6 coreceptor) and in-vitro experiments using MCF-7 breast cancer cells. Through the molecular docking analysis, the binding energies on the frizzled-8 receptor were -5.6, -5.1, -9.4, and -8.8 kcal/mol, respectively. Meanwhile, those on the LRP6 receptor, were -7.3, -6.2, -10.0, and -9.8 kcal/mol, respectively. The results showed that fucoidan had a favorable binding affinity for both receptors. Furthermore, it was discovered to reduce the interaction and binding affinity between Wnt agonists to frizzled-8 and LRP6 receptors. This reduction was reflected in the change in the binding energy of the fucoidan-Wnt agonist-frizzled 8 and fucoidan-Wnt agonist-LRP6 complexes, which exhibited decreases of -7.0 kcal/mol and -7.8 kcal/mol, respectively. Fucoidan was found stable in complexes with frizzled-8 receptor and co-receptor LRP6. ADMET study showed it's non-carcinogenic and can be distributed in the body. Fucoidan effectively inhibited ß-catenin production, a critical factor in the Wnt/ß-catenin pathway. The MCF-7 breast cancer cells were treated with fucoidan extract from S. echinocarpum at incubation times of 24, 48, and 72 h, resulting in a reduction of ß-catenin levels by 95.19%, 83.88%, and 80.88%, respectively. Fucoidan also shows no significant difference in value compared to fucoidan standard (F. vesiculosus) and doxorubicin. Fucoidan exhibited antiproliferative effects against breast cancer cells, specifically through its modulation of the Wnt/ß-Catenin pathway, and held great potential as an herbal anticancer agent.Communicated by Ramaswamy H. Sarma.

The Cortisol Levels, Histology, and Fine Structure of Various Tissues of Fish Gambusia affinis (Baird and Girard, 1853) after Exposure to Lead.

Background: Aquatic organisms demonstrate a high vulnerability to mortality when exposed to Pb, even at low concentrations. The objective of this investigation is to ascertain the histopathological alterations and cortisol concentrations in diverse tissues of Gambusia affinis, with a specific focus on the eggs and larvae, following exposure to varying concentrations of PbCl2. Methods: Adult specimens of G. affinis measuring 5-6 cm in length were obtained from a commercial fish breeding facility. A total of 8 fish with a 1 : 1 ratio of 4 pairs of broodstock were placed in an 8-liter aquarium. Following the adaptation phase, the broodstock underwent a spawning process that lasted for a duration of 7 days. Throughout the spawning process, assessments were conducted on the progression of the abdominal growth of the broodstock. Eggs ready to hatch and Gambusia larvae were taken and exposed to 0.1 mg/L PbCl2, 1 mg/L PbCl2, and control (without PbCl2) for 24 hours, with three replications. At the end of the experiment, histopathological analysis was conducted using the hematoxylin Ehrlich-eosin staining method and scanning electron microscopic (SEM) observation. The levels of Pb in gills were determined by employing atomic absorption spectrophotometer. The cortisol concentration in organ samples of fish was determined through the utilization of a cortisol ELISA Kit. Results: The findings of this investigation demonstrated an important bioaccumulation occurrence of Pb within the gills of Gambusia fish that were specifically subjected to 0.1 and 1 mg/L PbCl2. The histological structures of eggs and larvae that were subjected to PbCl2 exhibited impairment in comparison to the control group. The present study observed a significant elevation in cortisol levels among fish specimens that were subjected to PbCl2 exposure. Conclusions: The findings of this investigation suggest that the occurrence of Pb is linked to a rise in cortisol concentrations in various organs of G. affinis larvae. Furthermore, the research indicates that the exposure to Pb has a notable impact on the histological alterations in the eggs and larvae of Gambusia fish, implying that they are undergoing stress as a result of the Pb exposure.

Oxidative stress responses of microplastic-contaminated Gambusia affinis obtained from the Brantas River in East Java, Indonesia.

The biological impact of microplastic (MP) contamination on freshwater biota has sparked interest. Microplastics are thought to have a high potential for toxicity, affecting organism metabolism. The goal of this study was to investigating the antioxidant indicators on the gills and digestive tract of Gambusia affinis fish in the Brantas River reacted to microplastic pollution. The obtained data was evaluated using path analysis. The digestive tract had much greater levels of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) than the gill. SOD concentration in the gills was 13.7 ± 5.3 U/mL, while SOD concentration in the digestive tract was 16.3 ± 3.6 U/mL. The CAT concentration in the gills (5.3 ± 2.9 ng/mL) was higher than the CAT concentration in the digestive tract (10.5 ± 2.8) ng/mL, while the MDA concentration in the gills (690.8 ± 135.6 mU/mL) was higher than the MDA concentration in the digestive tract (869.6 ± 122.2) mU/mL. MP abundance has a direct effect on SOD and MDA in the gills. Meanwhile, the quantity of MP in the digestive tract has a direct effect on SOD and CAT, which affects the development of the MDA response.

Spatio-temporal patterns of occurrence of microplastics in the freshwater fish Gambusia affinis from the Brantas River, Indonesia.

The first investigation of microplastics in G. affinis from the Brantas River was carried out in this study. Microplastics were found at higher concentrations in gambusia fish captured downstream (209.18 ± 48.85 particles/gram) than upstream (24.44 ± 0.14 particles/gram). Microplastic particle concentrations in G. affinis have a positive linear relationship with fish length. The fiber was the most prominent shape at Sites 1 and 2, whereas the fragment was dominant at Sites 3 and 4. With a value of 45-48%, black dominates the entire site, followed by blue (29-38%), transparent colors (7-11%), red (2-4%), purple (1-3%), and other colors (5-7%). Microplastics measuring <0.1 mm are commonly found in fish bodies. Cyclohexylmethyl octyl ester (phthalic acid) is the most abundant component found in microplastics, accounting for 30.11% of the total. This study provides evidence that G. affinis can be used to monitor the presence of microplastic pollution in the Brantas River but further studies are needed regarding the effects of microplastics and their health hazards on fish.

Marine and brackish Luticola D.G.Mann (Bacillariophyta) species from the Java Sea and South China Sea coasts with the description of three new species.

In this study, samples were collected from the Java Sea coasts, from the South China Sea in Hainan Island coasts and Quang Yên region and Rú Chá mangrove near Hue in Central Vietnam. In studied samples a total of eight Luticola species have been observed. Three of the taxa studied are described herein as species new to science - Luticolaorientalis sp. nov., L.cribriareolata sp. nov. and L.halongiana sp. nov. Under light microscopy (LM) L.orientalis sp. nov. and L.cribriareolata sp. nov. are similar with rhombic-lanceolate to rhombic/ elliptic-lanceolate to elliptic valve shapes and narrowly rounded apices. Both species can be easily distinguished by stria density (higher density in L.orientalis). Under SEML.cribriareolata is characterized by cribrate areola occlusions, a character thus far observed only in three established species. The remaining species of the whole genus known thus far are characterized by hymenate areola. Similar morphology Luticola species have been observed from tropical mangrove forests from Madagascar but they all can be easily distinguished based on the lack of grooves in the central area. The third species - L.halongiana sp. nov. has rhombic-elliptic to rhombic-lanceolate valves with broadly rounded to slightly protracted apices in larger specimens. This species has a relatively broad central area. Also unique among brackish-water Luticola is the small, rounded stigma positioned almost midway between the valve center and valve margin. In the habitats from which the new species are described we also identified five established Luticola taxa including, L.belawanensis, L.celebesica, L.inserata, L.seposita and L.tropica. For those species we provide detailed SEM characteristics of valve ultrastructure, as well as the range of environmental conditions and geographic distribution within the study area.

Genotoxicity in the rivers from the Brantas catchment (East Java, Indonesia): occurrence in sediments and effects in Oreochromis niloticus (Linnæus 1758).

This paper reports the first data from an integrated study investigating genotoxicity in the Brantas River, Java, Indonesia. Results showed that organic sediment extracts from the sites in the Brantas Delta retained genotoxic compounds identified using the SOS Chromotest and that the Aloo River and, to a lesser extent, the Surabaya River were the most contaminated studied sites. This genotoxicity was attributable to compounds that did not require any bioactivation under the test conditions. Occurrence of genotoxic effects was further investigated in erythrocytes from Nile tilapia, Oreochromis niloticus. High numbers of micronuclei were counted, especially in fish sampled in the rivers of the Brantas Delta. Moreover, cytoplasmic alterations which could be indicative of the presence of lipofuscin were found in the cytoplasm of the fish blood cells, especially in fish from the Aloo, Surabaya and Kalimas rivers. Altogether, our data showed that genotoxicity is occurring in fish living in rivers of the delta of the Brantas River and suggest that sediments from these sites may constitute a major source of pollution and hazard for species living or feeding in the area.

Toward Sustainable Environmental Quality: Priority Research Questions for Asia.

Environmental and human health challenges are pronounced in Asia, an exceptionally diverse and complex region where influences of global megatrends are extensive and numerous stresses to environmental quality exist. Identifying priorities necessary to engage grand challenges can be facilitated through horizon scanning exercises, and to this end we identified and examined 23 priority research questions needed to advance toward more sustainable environmental quality in Asia, as part of the Global Horizon Scanning Project. Advances in environmental toxicology, environmental chemistry, biological monitoring, and risk-assessment methodologies are necessary to address the adverse impacts of environmental stressors on ecosystem services and biodiversity, with Asia being home to numerous biodiversity hotspots. Intersections of the food-energy-water nexus are profound in Asia; innovative and aggressive technologies are necessary to provide clean water, ensure food safety, and stimulate energy efficiency, while improving ecological integrity and addressing legacy and emerging threats to public health and the environment, particularly with increased aquaculture production. Asia is the largest chemical-producing continent globally. Accordingly, sustainable and green chemistry and engineering present decided opportunities to stimulate innovation and realize a number of the United Nations Sustainable Development Goals. Engaging the priority research questions identified herein will require transdisciplinary coordination through existing and nontraditional partnerships within and among countries and sectors. Answering these questions will not be easy but is necessary to achieve more sustainable environmental quality in Asia. Environ Toxicol Chem 2020;39:1485-1505. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Screening of NO Inhibitor Release Activity from Soft Coral Extracts Origin Palu Bay, Central Sulawesi, Indonesia.

BACKGROUND: As a marine organism, soft corals can be utilized to be various bioactive substances, especially terpenoids and steroids. The soft corals family which produces bioactive generally come from clavulariidae, alcyoniidae, nephtheidae and xeniidae family. OBJECTIVE: To investigate the bioactivity of Nitric Oxide (NO) inhibitor release from soft coral crude extracts of Sinularia sp. (SCA), Nephthea sp. (SCB), Sarcophyton sp. (SCC), Sarcophyton sp. (SCD), Sinularia sp. (SCE) and Sinularia sp. (SCF). MATERIALS AND METHODS: Soft coral is collected from Palu Bay (Central Sulawesi). NO inhibitory release activity measured according to the Griess reaction. Soft corals sample macerated with 1:2 (w/v). Then, Soft coral extracts with the best NO Inhibitor activity partitioned with Dichloromethane, Ethyl acetate, and n-butanol. The bioactive of all crude extracts were identified by GC-MS to find compounds with anti-inflammatory potential. RESULTS: Sarcophyton sp. (SCC) and Sinularia sp. (SCF) are able to inhibit NO concentrations of 0.22 ± 0.04 and 0.20 ± 0.04 µM at 20 mg/mL, respectively. The chemical constituents determined and showed the potential as anti-inflammatory in the crude of Sinularia sp. (SCA) were Octacosane (3.25%). In Nephthea sp., (SCB) were Cyclohexene, 6-ethenyl-6- methyl-1-(1-methylethyl)-3-(1-methylethylidene)-,(S)- (0.55%); Azulene, 1,2,3,4,5,6,7,8- octahydro-1,4-dimethyl-7-(1-methylethylidene)-, (1S-cis)- (0.53%); and 1,7,7-Trimethyl- 2-vinylbicyclo[2.2.1]hept-2-ene (4.72%). In Sarcophyton sp, (SCC) were Eicosane (0.12%); Nonacosane (10.7%); 14(ß)-Pregnane (0.87%); Octacosane 6.39%); and Tricosane (1.53%). In Sarcophyton sp. (SCD) were 14(ß)-Pregnane (2.69%); and Octadecane (27.43%). In crude of Sinularia sp. (SCE) were Oleic Acid (0.63%); 7,10-Hexadecadienoic acid, methyl ester (0.54%); 14(ß)-Pregnane (1.07%); 5,8,11,14-Eicosatetraenoic acid, ethyl ester, (all-Z)- (4.60%); Octacosane (7.75%); and 1,2-Benzisothiazole, 3-(hexahydro-1Hazepin- 1-yl)-, 1,1-dioxide (1.23%). In the crude of Sinularia sp., (SCF) were Oxirane, decyl- (1.38%); Nonacosane (0.57%); Cyclohexanol, 5-methyl-2-(1-methylethenyl)- (0.61%); 14B-Pregnane (0.76%); and Tetratriacontane (1.02%). CONCLUSION: The extract of Sarcophyton sp. (SCC) and Sinularia sp. (SCF) showed the best NO inhibitory release activity. This study is making soft corals from Central Sulawesi, Indonesia can become a potential organism in the discovery and development of bioactive substances anti-inflammatory.

Cellular immune responses and phagocytic activity of fishes exposed to pollution of volcano mud.

Since May 29, 2006, a mud volcano in the Brantas Delta of the Sidoarjo district has emitted mud that has inundated nearby villages. Pollution in this area has been implicated in detrimental effects on fish health. In fishes, leukocyte and phagocytic cells play a vital role in body defenses. We report for the first time the effect of "LUSI" volcano mud on the immune systems of fish in the Brantas Delta. The aim of this study was to find biomarkers to allow the evaluation of the effects of volcanic mud and anthropogenic pollution on fish health in the Brantas Delta. The study took places at the Brantas Delta, which was polluted by volcano mud, and at reference sites in Karangkates and Pasuruan. Leukocyte numbers were determined using a Neubauer hemocytometer and a light microscope. Differential leukocyte counts were determined using blood smears stained with May Grunwald-Giemsa, providing neutrophil, lymphocyte and monocyte counts. Macrophages were taken from fish kidney, and their phagocytic activity was measured. In vitro analyses revealed that leukocyte and differential leukocyte counts (DLC) were higher in Channa striata and Chanos chanos caught from the polluted area. Macrophage numbers were higher in Oreochromis mossambicus than in the other species, indicating that this species is more sensitive to pollution. In areas close to volcanic mud eruption, all specimens had lower phagocytic activity. Our results show that immune cells were changed and phagocytic activity was reduced in the polluted area indicating cytotoxicity and alteration of the innate immune system in fishes exposed to LUSI volcano mud and anthropogenic pollution.