Ingestion and impacts of water-borne polypropylene microplastics on Daphnia similis.

Polypropylene microplastics are the leading contaminant in aquatic environments, although research on their toxicity remains scarce. The proposed research focuses on the harmful consequences of acute exposure to polypropylene microplastics in Daphnia similis. This work converts widely available polypropylene bags into microplastics using xylene. FTIR findings demonstrated the lack of xylene residue in the produced polypropylene microplastic particles, which were spherical and ranged in size from 11.86 to 44.62 µm (FE-SEM). The results indicate that acute exposure to polypropylene microplastics causes immobility in D. similis. Ingestion of microplastics enhances the generation of reactive oxygen species (ROS), as shown by biochemical studies. Due to the production of free radicals in D. similis, the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) and a non-antioxidant enzyme of reduced glutathione (GSH) and also oxidative stress effects in lipid (lipid peroxidation - LPO), protein (carbonyl protein - CP) were increased. Additionally, the amount of the neurotransmitter enzyme acetylcholinesterase (AChE) activity was decreased. These findings indicate that the accumulation of polypropylene microplastics in the bodies of filter-feeding organisms should aggravate toxicity in the freshwater environment.

Comparative toxicity of silver nanoparticles and silver nitrate in freshwater fish Oreochromis mossambicus: A multi-biomarker approach.

Silver nanoparticles (AgNPs) in the aquatic environment affect ecological repercussions and have fatal impacts on aquatic animals. The current study examined and correlated the toxicity of silver nitrate (AgNO3) and silver nanoparticles (AgNPs) to the Mozambique tilapia, Oreochromis mossambicus. The comparative toxicity studies were done by exposing O. mossambicus to various doses of AgNO3 and AgNPs (0, 25, 50, 75, and 100 µg/L) over a 7-day subacute exposure period. AAS analysis was used to detect Ag accumulation, while the histological examination established gill tissue damage. Oxidative stress affects lipid peroxidation (LPO) and protein carbonyl activity (PCA) in the gill tissue. Antioxidant parameters such as glutathione-S-transferase (GST), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase activity (CAT), and non-enzymatic antioxidants such as metallothionein (MT) and reduced glutathione. The serum in the blood was used to determine non-specific immunological characteristics such as lysozyme (LYZ), myeloperoxidase (MPO), and respiratory burst activity (RBA). The neurotoxic impact of acetylcholine esterase activity (AChE) was investigated in brain tissues. The findings demonstrated that larger concentrations of AgNO3 than AgNPs improved enzymatic antioxidant activities in the gill tissue. Histological examination of fish gills demonstrated that both AgNPs and AgNO3 induced telangiectasia and epithelial cell hyperplasia. By increasing the concentration of AgNPs and AgNO3, the present research demonstrated that silver accumulation leads to inefficient oxidative stress and altered enzymatic and non-enzymatic parameters, leading to cellular damage.

Dietary consumption of polypropylene microplastics alter the biochemical parameters and histological response in freshwater benthic mollusc Pomacea paludosa.

One of the most common environmental pollutant in aquatic ecosystems are polypropylene microplastics and their impacts on aquatic organisms are still scarce. The study aimed to prepare polypropylene microplastics using organic solvent (spherical and 11.86-44.62 µm) and then test their toxicity on the freshwater benthic mollusc grazer Pomaceae paludosa. The present study investigated chronic (28 days) exposure of polypropylene microplastics via dietary supplements (250 mg kg-1, 500 mg kg-1 & 750 mg kg-1) in P. paludosa, and the toxic effect was evaluated in digestive gland tissue. The FTIR results revealed no change in polypropylene microplastics during ingestion or after egestion. On the other hand, Ingestion causes accumulation in their bodies and disrupts redox homeostasis. Meanwhile, alteration occurs in oxidative stress-related biomarkers such as increased reactive oxygen species level (ROS), impaired the biochemical parameters of antioxidant system catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), and glutathione - S- transferase (GST), deterioration of oxidative stress effects in lipid peroxidation (LPO) and carbonyl protein (CP) and changed the digestive enzymes such as amylase, pepsin, esterase and alkaline phosphatase that are measured in hepatopancreas tissue. The histology results revealed that ingesting these microplastics caused severe damage to the digestive gland cells. According to the findings, ingestion of polypropylene microplastics in benthic freshwater mollusc causes more serious harm and impacts energy acquisition. This finding represents the ecological risk of polypropylene microplastic pollution in the freshwater ecosystem.

Protective effects of dietary supplementation of probiotic Bacillus licheniformis Dahb1 against ammonia induced immunotoxicity and oxidative stress in Oreochromis mossambicus.

The goal of this study was to assess the efficacy of probiotics in mitigating ammonia-induced toxicity in fish. Fish were divided into four groups: control, only probiotic, only ammonia, and combined ammonia + probiotic. For 8 weeks, the Oreochromis mossambicus were exposed to waterborne ammonia at 1.0 mg L-1 and/or dietary Bacillus licheniformis Dahb1 at 107 cfu g-1. After the 4th and 8th weeks, the fish were evaluated for growth performance, enzymatic and non-enzymatic antioxidant activities (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) reduced glutathione (GSH), neurotoxicity (acetylcholinesterase - AChE), non-specific immune responses (lysozyme (LYZ), myeloperoxidase (MPO), reactive nitrogen and oxygen species (RNS and ROS) and oxidative stress effects (lipid peroxidation (LPO), DNA damage)). Our results showed that in the absence of waterborne ammonia exposure, B. licheniformis Dahb1 significantly improved growth performance, enzymatic and non-enzymatic antioxidant capacity, AChE activity, non-specific immune response and decreased oxidative stress effect. Ammonia exposure resulted in significantly lower growth performance, reduced enzymatic and non-enzymatic antioxidant ability, decreased AChE activity, decreased non-specific immune response and increased oxidative stress effect. When O. mossambicus were exposed to ammonia, supplementation with B. licheniformis Dahb1 in the diet significantly increased survival, indicating that it may have a significant protective effect against ammonia toxicity by enhancing enzymatic and non-enzymatic antioxidant ability, activity of AChE, non-specific immune response and reduced oxidative stress effect. According to our findings, diet supplementation of B. licheniformis Dahb1 (107 cfu g-1) has the potential to combat ammonia toxicity in O. mossambicus.

The dietary supplementation of zinc oxide and selenium nanoparticles enhance the immune response in freshwater fish Oreochromis mossambicus against aquatic pathogen Aeromonas hydrophila.

BACKGROUND: Green nanoparticles are subjected as an immunostimulant against bacterial pathogens. METHODS: Murraya koenigii berry extract-based synthesized zinc oxide nanoparticles (Mb-ZnO NPs) and selenium nanoparticles (Mb-Se NPs) were relatively analyzed for immunostimulation in serum and mucus fish Oreochromis mossambicus against Aeromonas hydrophila infections. Initial minimum inhibitory concentration (MIC) was determined for both Mb-ZnO NPs and Mb-Se NPs followed by specific growth rate (SGR), antioxidant level (Superoxide dismutase activity (SOD), Catalase activity (CA), and Glutathione peroxidase activity (GPx)), and immune parameters Myeloperoxidase activity (MPO), Respiratory burst activity (RBA), Lysozyme activity (LYZ), Alkaline phosphatase activity (ALP), Serum antiprotease activity and Natural complement activity (NAC). RESULTS: The potential bacterial inhibition property of Mb-ZnO NPs and Mb-Se NPs exhibited the most negligible concentration of 25 and 15 µg mL-1, respectively, against A. hydrophila. In addition, Mb-ZnO NPs and Mb-Se NPs exhibited 70-80 % and 90-95 % diminished biofilm activity at 50 µg mL-1 that was viewed under an inverted research microscope and confocal laser scanning microscopy (CLSM). Protein leakage and nucleic acid leakage assay quantified oozed out protein and nucleic acid from A. hydrophila that confirms Mb-Se NPs exhibited vigorous antibacterial activity than Mb-ZnO NPs at tested concentrations. Oreochromis mossambicus fed with Mb-ZnO NPs and Mb-Se NPs supplemented diet at different concentrations (0.5 mg/kg, 1 mg/kg and 2 mg/kg) improved SGR along with a rise in the immune response of those fishes against A. hydrophila infection. Serum and mucus of fish fed with Mb-Se NPs supplemented diet exhibited a significant rise in antioxidant level SOD, CA and GPx at a dosage of 2 mg/kg. Likewise, lipid peroxidation assay detected significantly diminished oxidative stress in the serum and mucus of fish fed with Mb-Se NPs supplemented diet (2 mg/kg). Enhanced immune parameters in serum and mucus of fish fed with Mb-Se NPs supplemented diet determined by MPO, RBA, LYZ, ALP, Serum antiprotease activity and NAC. CONCLUSION: Thus O. mossambicus fed with Mb-Se NPs supplemented diet was less prone to become infected by aquatic pathogen A. hydrophila established by challenge study. On the whole, Mb-Se NPs supplemented diet ensured the rise in antioxidant response that boosts the immune responses and reduces the chance of getting infected against A. hydrophila infections.

ß-1,3-Glucan binding protein-based silver nanoparticles enhance the wound healing potential and disease resistance in Oreochromis mossambicus against Aeromonas hydrophilla.

Here we attempted to synthesize ß-1,3-glucan binding protein-based silver nanoparticles (Phß-GBP-AgNPs) and evaluate its wound healing and disease resistance prompting ability in Oreochromis mossambicus. Using a column chromatography technique, an immune molecule, Phß-GBP was purified from the haemolymph of rice field crab, Paratelphusa hydrodromus. Phß-GBP-AgNPs were synthesized and described through SDS-PAGE, UV-vis spectroscopy, HR-TEM, XRD and FTIR analysis. HR-TEM revealed that the synthesized Phß-GBP-AgNPs were spherical with a 20-40 nm size range and the particles were not aggregated. Wound and infection were experimentally generated in O. mossambicus and treated with Phß-GBP, chem-AgNPs and Phß-GBP-AgNPs for 20 days. The immune parameters (peroxidase, lysozyme and protease) and antioxidant enzymes (SOD and catalase) were examined in the serum of experimental fish. Phß-GBP-AgNPs elevated the immune and antioxidant enzymes during the healing process and enhanced the wound healing percentage in fish than Phß-GBP and chem-AgNPs. The immune parameters and antioxidant enzymes were declined in the serum of fish (treated with Phß-GBP-AgNPs) after the mid-period of wound healing. Compared to others, relative percentage survival was increased in experimentally wounded and infected fish treated with Phß-GBP-AgNPs against Aeromonas hydrophila. Moreover, Phß-GBP-AgNPs exhibited less toxicity towards Artemia salina than chem-AgNPs during 24 h exposure period. As a result, Phß-GBP-AgNPs may act as an alternative to commercial antibiotics and be considered an effective immunostimulant in treating skin lesions in intensive farming.

Interactive effects of freshwater acidification and selenium pollution on biochemical changes and neurotoxicity in Oreochromis mossambicus.

Effect of selenium and acidification in freshwater environment was assessed solitary but no reports are available on the impacts of both factors act together. In the present study, effects of combined simultaneous exposure to selenium (Se) and low pH were assessed in Mozambique tilapia, Oreochromis mossambicus. Responses were measured based on antioxidant defenses (enzymatic SOD, CAT, GPx and non-enzymatic GSH), biotransformation enzyme (GST), metallothionein levels (MT), oxidative damage (LPO, CP), Na+/K+-ATPase (NKA) activity in gills and liver tissues and neurotoxicity (acetylcholinesterase, AChE) response in brain tissue. Fish were exposed to combined treatment at different pH levels (7.5, control (optimum pH for tilapia growth); 5.5, low pH) and Se concentrations (0, 10, and 100 µg L-1). Toxicity levels of Se were not significantly different under control and low pH indicating that pH did not affect Se toxicity. Levels of GSH and MT were enhanced in Se-exposed fish at both pH. Combined effects of high Se concentration and low pH decreased SOD and CAT activities and increased those of GPx and GST. However, organisms were not able to prevent cellular damage (LPO and CP), indicating a condition of oxidative stress. Furthermore, inhibition of Na+/K+-ATPase activity was showed. Additionally, neurotoxicity effect was observed by inhibition of cholinesterase activity in organisms exposed to Se at both pH conditions. As a result, the combined stress of selenium and freshwater acidification has a slight impact on antioxidant defense mechanisms while significantly inhibiting cholinesterase and Na+/K + -ATPase activity in fish. The mechanisms of freshwater acidification mediating the toxic effects of trace non-metal element on freshwater fish need to investigate further.

ß-glucan extracted from eukaryotic single-celled microorganism Saccharomyces cerevisiae: Dietary supplementation and enhanced ammonia stress tolerance on Oreochromis mossambicus.

Ammonia is a widespread pollutant that is toxic to living organisms in aquaculture. This study aimed to evaluate the effects of a diet supplemented with beta-glucan from yeast, Saccharomyces cerevisiae (Sc-ßG), on the stress response of Oreochromis mossambicus (Tilapia) to ammonia. Fish were divided into four groups, including a control fed a basal diet and three experimental groups fed diets supplemented with Sc-ßG at 2, 5 and 10 mg/g respectively. After 8 weeks, experimental groups were exposed to ammonia at 100 mg L-1 for 1 week. Growth was measured after the 8-week feeding trial and serum, mucus, and liver tissue were sampled before and after the ammonia challenge. Compared with the control diet, feed supplemented with Sc-ßG at 10 mg/g significantly (p < 0.05) improved growth performance (7.8-9.9 g increase in weight). The cellular immune responses (myeloperoxidase, reactive oxygen species, and reactive nitrogen species), humoral immune responses (alkaline phosphatase, lysozyme, and peroxidase inhibition), and antioxidant response (catalase, superoxide dismutase, and glutathione) were tested in serum, mucus and liver tissue. Compared with the control, these responses were significantly (p < 0.05) enhanced at 10 mg/g supplementation with Sc-ßG. This study demonstrates that Sc-ßG may be applied to induce stress tolerance and improve growth performance in aquaculture.

Chronic exposure of Oreochromis niloticus to sub-lethal copper concentrations: Effects on growth, antioxidant, non-enzymatic antioxidant, oxidative stress and non-specific immune responses.

BACKGROUND: Trace elements of copper (Cu) are one of the main forms of ecological noxious waste in freshwater systems that affect the survival and development of organisms. The objective of the current study was to investigate the effects of chronic exposure to Cu on the growth, oxidative stress, immune and biochemical response in the Nile tilapia, Oreochromis niloticus. METHODS: Three groups of O. niloticus were tested as follows; the first group was used as the control (not treated with Cu in water), while the 2nd and 3rd groups were exposed to (low) 40 µg L-1 and (high) 400 µg L-1 concentrations of Cu added to water, respectively. The duration of the experiment, which was conducted in triplicate, was 60 d. End points were evaluated on days 30 and 60. Following 30 d and 60 d of exposure to Cu, the fish were removed from experimental tanks to determine growth. Consequently, blood samples were collected from caudal veins at the end of the trial period (30 d and 60 d) and serum was separated to evaluate different immunological parameters, such as lysozymes (LYZ), respiratory burst activity (RBA) and myeloperoxidase (MPO). Gill and liver tissues were collected for evaluation of Cu and certain biochemical parameters as follows: antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST); non-enzymatic antioxidants such as glutathione (GSH) and metallothionein (MT), and oxidative stress indicators such as malondialdehyde (MDA) and protein carbonyl (PCO). The results pertaining to treatments and the control were compared using two-way ANOVA and Tukey's HSD test. The level of significance was set at P ≤ 0.05. Data were expressed as mean ±â€¯SD. RESULTS: Chronic exposure to Cu did not induce any mortality in fish during the test period. However, following exposure to Cu, growth of fish in the exposed groups was affected more than that in the control group (unexposed to Cu). In addition, accumulation of Cu in the liver tissue was higher than that in the gill tissues of fish exposed to Cu, compared to that in the control. Gill and liver tissues of Cu-exposed fish showed a significant (P ≤ 0.05) reduction in the activities of the antioxidant enzymes, SOD, CAT, GPx, and GST, compared to those of unexposed fish. Non-enzymatic antioxidants, GSH and MT, in gill and liver tissues were significantly increased (P ≤ 0.05) in fish exposed to both concentrations of Cu, compared to those in unexposed fish. Oxidative stress indicators, MDA and PCO in gills and liver of Cu-exposed fish was significantly (P ≤ 0.05) at both tested concentrations, when compared to control group. Non-specific immune response of LYZ, RBA, and MPO activity in serum decreased significantly (P ≤ 0.05) in Cu-exposed fish, compared with that of unexposed fish. CONCLUSION: Overall, the present results highlighted that chronic exposure to Cu ions may exert a strong effect on the antioxidant and immune responses of O. niloticus. Changes in antioxidant enzymes, oxidative stress effects and immune parameters during post-chronic metal exposure may indicate the potential of these parameters as biomarkers of metal toxicity in aquatic ecosystems.

Growth inhibition and antibiofilm potential of Ag nanoparticles coated with lectin, an arthropod immune molecule.

Lectins from the haemolymph of arthropods, including crustaceans, are molecules potentially involved in the immune recognition and phagocytosis. Here, lectin was purified from the haemolymph of blue swimmer crab Portunus pelagicus, using mannose-coupled sepharose CL-4B affinity column chromatography. In SDS-PAGE analysis, lectin showed a molecular mass of approximately 155kDa. The synthesis of lectin-coated silver nanoparticles (lectin-coated AgNPs) was confirmed by UV-Vis spectroscopy, XRD, FTIR, SEM, TEM, SAED, and EDX analysis. TEM analysis revealed that lectin-coated AgNPs were spherical in shape with size of 30-57nm. Their antibacterial activity against human pathogenic Gram negative and Gram-positive bacteria was determined by agar well diffusion method. Lectin-coated AgNPs showed significant antimicrobial activity when compared to lectin and silver nitrate tested alone. The antibiofilm properties of lectin-coated AgNPs were also investigated on human pathogenic Gram-negative Proteus vulgaris, Pseudomonas aeruginosa and Gram-positive Enterococcus faecalis and Bacillus pumilus. Lectin-coated AgNPs showed antibiofilm activity on the bacteria as well as on Candida albicans. Lectin-coated AgNPs reduced the biofilm architecture interfering with cell adhesion and polysaccharide matrix. This was additionally confirmed by exopolysaccharide (EPS) quantification index revealing the trouble in the structural reliability of biofilm by decrease in EPS and bacterial adhesion to hydrocarbons.