Bacterial screening in Indian coastal regions for efficient polypropylene microplastics biodegradation.

Polypropylene based medical devices significantly increased production and usage in COVID-19 pandemic states, and this material is very resilient in the environment. Thus, more than ever, rapid action is needed to reduce this pollution. This study focuses on the degradation of polypropylene microplastics (PP MPs) by unique marine bacterial strains obtained from the Thoundi (Bacillus tropicus, Bacillus cereus, Stenotrophomonas acidaminiphila, and Brucella pseudintermedia) and Rameshwaram coasts (Bacillus cereus). Those above five bacterial strains were chosen after preliminary screening of their hydrophobicity, biofilm-forming capabilities, and responsiveness to the zone of clearance technique. During the biodegradation process (28 days), the growth, metabolic activity, and viability of these five isolates were all raised. After the post-biodegradation process, the weight loss percentages of the mentioned bacterial strains treated with PP MPs gradually decreased, with values of 51.5 ± 0.5 %, 47.5 ± 0.5 %, 33 ± 1 %, 28.5 ± 0.5 and 35.5 ± 0.5 %, respectively. UV-Vis DRS and SEM analysis confirmed that bacterial strains adhering to MPs cause cracks and cavities on their surface. The degradation of PP MPs can be inferred from alterations in the FT-IR spectrum, specifically in the carbonyl group range of 1100-1700 cm-1, as well as changes in the 1H NMR spectrum, including chemical shift and proton peak pattern alterations. Bacterial strains facilitated the degradation of PP MPs through the secretion of hydrolase-categorized enzymes of protease, lipase, and esterase. The findings of this study indicate that marine bacteria may possess distinctive characteristics that facilitate the degradation of plastic waste and contribute to environmental conservation.

Combined toxic effects of environmental predominant microplastics and ZnO nanoparticles in freshwater snail Pomaceae paludosa.

In the past few years, microplastics are one of the ubiquitous threatening pollutants in aquatic habitats. These persistent microplastics interact with other pollutants, especially nanoparticles were adherent on the surface, which causes potential hazards in the biota. In this study, the toxic effects of individual and combined (28 days) exposure with zinc oxide nanoparticles and polypropylene microplastics were assessed in freshwater snail Pomeacea paludosa. After the experiment, the toxic effect was evaluated by the estimation of vital biomarkers activities including antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), oxidative stress in carbonyl protein (CP), lipid peroxidation (LPO), and digestive enzymes (esterase and alkaline phosphatase). Chronic exposure to pollutants in snails causes increased reactive oxygen species level (ROS) and generates free radicals in their body which leads to impairment and alterations of biochemical markers. Where alteration in acetylcholine esterase (AChE) activity and decreased digestive enzymes (esterase and alkaline phosphatase) activities were observed in both individual and combined exposed groups. Further, histology results revealed the reduction of haemocyte cells, the disintegration of blood vessels, digestive cells, calcium cells, and DNA damage was also detected in the treated animals. Overall, when compared to individual exposures, combined exposure of pollutants (zinc oxide nanoparticles and polypropylene microplastics) causes more serious harms including decline and increased antioxidant enzyme parameters, damage the protein and lipids by oxidative stress, increased neurotransmitter activity, decrease digestive enzyme activities in the freshwater snail. The outcome of this study concluded that polypropylene microplastics along with nanoparticles cause severe ecological threats and physio-chemical effects on the freshwater ecosystem.

Biochemical, Genotoxic and Histological Implications of Polypropylene Microplastics on Freshwater Fish Oreochromis mossambicus: An Aquatic Eco-Toxicological Assessment.

In recent years, polypropylene microplastic has persisted in freshwater ecosystems and biota, forming ever-growing threats. This research aimed to prepare polypropylene microplastics and evaluate their toxicity to the filter feeder Oreochromis mossambicus. In this research, fish were given a dietary supplement of polypropylene microplastics at 100, 500, and 1000 mg/kg for acute (96 h) and sub-acute (14 days) durations to assess toxic effects on liver tissues. FTIR results revealed the presence of polypropylene microplastic in their digestion matter. The ingestion of microplastics in O. mossambicus led to fluctuations in homeostasis, an upsurge in reactive oxygen species (ROS) levels, an alteration in antioxidant parameters, including superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), and glutathione peroxidase (GPx); a promotion in the oxidation of lipid molecules; and a denaturation in the neurotransmitter enzyme acetylcholinesterase (AChE). Our data indicated that sustained exposure to microplastics (14 days) produced a more severe threat than acute exposure (96 h). In addition, higher apoptosis, DNA damage (genotoxicity), and histological changes were found in the liver tissues of the sub-acute (14 days) microplastics-treated groups. This research indicated that the constant ingestion of polypropylene microplastics is detrimental to freshwater environments and leads to ecological threats.

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.

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.

Toxicity evaluation of polypropylene microplastic on marine microcrustacean Artemia salina: An analysis of implications and vulnerability.

Polypropylene microplastic particles are one of the predominant pollutants in marine ecosystems and their toxic effects are unknown in aquatic biota. The study aims to prepare the spherical shaped polypropylene microplastics (size range 11.86 µm-44.62 µm) and assess their toxic effects (1, 25, 50, 75 and 100 µg/mL) in various life stages (nauplii, metanauplii and juvenile) of marine microcrustacean Artemia salina within 48 h. In addition, microplastics ingestion by Artemia nauplii was proved by FTIR analysis. The results revealed, microplastics accumulation in their tract leads to change in their homeostasis, as followed increase in the oxidative burst causes mortality in nauplii (LC50 40.947 µg/mL) and meta nauplii (LC50 51.954 µg/mL). In juvenile, swimming behaviour was changed. Moreover, microplastic consumption disturbs the antioxidant biomarkers such as superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione -S- Transferase (GST) and reduces the neurotransmitter enzyme acetylcholinesterase (AChE) activity. In addition, histology of juvenile Artemia showed damage in epithelial cells. This study indicates that exposure to polypropylene microplastics is more harmful to zooplanktonic organisms of the marine ecosystem.

Synthesis and physicochemical characteristics of Ag-doped hydroxyapatite nanoparticles, and their potential biomedical applications.

In this present scenario, hydroxyapatite (HAp) nanostructures were synthesized through green routes for biomedical applications, particularly remediation towards human pathogens and cancer cells. The present study aims at forming non-toxic and eco-friendly silver (Ag+) doped HAp using Polyethylene glycol (PEG), Cetyl Trimethyl ammonium bromide (CTAB) and curcumin. Ag+ doped HAp nanoparticles (NPs) were prepared by the sol-gel method with a cube and rod-like morphology. Ag-HApNPs showed a sharp and well-defined diffraction peak, which possesses the hexagonal crystalline structure with space group P63/m. The Fourier-transform infrared spectroscopy and Raman spectra confirmed the formation of Ag-HApNPs, and the bandgap values were obtained using UV-DRS analysis. The Ag-HApNPs with PEG, CTAB and curcumin might be fabricated materials were examined against antibacterial, antifungal, antioxidant, and cytotoxic activities, which provided exemplary biomedical applications. Overall, Ag-HApNPs can be used as potential drug delivery and perspectives to control multidrug-resistant pathogens.

Comprehensive Analysis of SARS-COV-2 Drug Targets and Pharmacological Aspects in Treating the COVID-19.

Corona viruses are enveloped, single-stranded RNA (Ribonucleic acid) viruses, and they cause pandemic diseases having a devastating effect on both human healthcare and the global economy. To date, six corona viruses have been identified as pathogenic organisms, which are significantly responsible for the infection and cause severe respiratory diseases. Among them, the novel SARS-CoV-2 (Severe Acute Respiratory Syndrome coronavirus 2) caused a major outbreak of coronavirus diseases in 2019 (COVID-19). Coronaviridae family members can affect both humans and animals. In humans, coronaviruses cause a severe acute respiratory syndrome with mild to severe outcomes. Several structural and genomics aspects have been investigated, and the genome encodes about 30 proteins most of them with unknown function though they share remarkable sequence identity with other proteins. There are no potent drugs against SARS-CoV-2 and several trials are underway to investigate the possible therapeutic agents against viral infection. However, some of the antiviral drugs that have been investigated against SARS-CoV-2 are under clinical trials. In the current review, we comparatively emphasize the emergence and pathogenicity of the SARS-CoV-2 and their infection, and discuss the various putative drug targets of both viral and host receptors for developing effective vaccines and therapeutic combinations to overcome the viral outbreak.

Antibiofilm and anticancer potential of ß-glucan-binding protein-encrusted zinc oxide nanoparticles.

ß-Glucan-binding protein (ßGBP) is important for the rational expansion of molecular biology. Here, zinc oxide nanoparticle (ZnONP) was synthesized using ßGBP from the crab Scylla serrata (Ss-ßGBP-ZnONP). Ss-ßGBP-ZnONP was observed as a 100 kDa band on sodium dodecyl sulfate polyacrylamide gel and characterized with UV-vis spectroscopy at 350 nm. X-ray diffraction analysis displayed values consistent with those for zincite. Fourier transform infrared spectroscopy revealed the presence of functional groups, including amide, alcohol, alkane, alkyl halide, and alkene groups. The zeta potential (-5.36 mV) of these particles indicated their stability, and transmission electron microscopy revealed the presence of 50 nm nanocones. Ss-ßGBP-ZnONPs were tested at 100 µg/mL against the gram-positive Enterococcus faecalis and gram-negative Pseudomanas aeruginosa using confocal laser scanning microscopy and the bacterial viability assay was also performed. The growth of MCF7 breast cancer cells was inhibited following treatment with 75 µg/mL Ss-ßGBP-ZnONPs. Thus, Ss-ßGBP-ZnONPs have the ability to control the growth of pathogenic bacteria and inhibit the viability of MCF7 breast cancer cell lines.

Anti-biofilm properties and immunological response of an immune molecule lectin isolated from shrimp Metapenaeus monoceros.

The study is carried out to understand the antimicrobial and immunological response of a potential immune molecule lectin, MmLec isolated from haemolymph of Speckled shrimp, Metapenaeus monoceros. MmLec was purified using mannose coupled Sepharose CL-4B affinity chromatography, which was further subjected on SDS-PAGE to ascertain the distribution of their molecular weight. Sugar binding specificity assay was conducted at various pH and temperatures to investigate the binding affinity of MmLec towards the specific carbohydrate molecule. Functional analysis of immune molecule MmLec included haemagglutination assays performed using human erythrocytes and yeast agglutination activity against Saccharomyces cerevisiae which, were analyzed using light microscopy. In order to study the antimicrobial activity, two Gram-negative (Vibrio parahaemolyticus and Aeromonas hydrophila) and two Gram-positive (Staphylococcus aureus and Enterococcus faecalis) bacteria were treated with purified MmLec. Moreover, these bacterial species were also treated at different concentration of the MmLec to speculate the antibiofilm properties of MmLec which was analyzed under Light Microscopy and Confocal Laser Scanning Microscopy. In addition, other functional characterization of MmLec showed the uniqueness of MmLec in agglutination of human erythrocyte as well as the cells of yeast Saccharomyces cerevisiae. Also, the phenoloxidase activity and encapsulation assay was evaluated. MTT assay displayed that MmLec are potent in anticancer activity. The study will help to understand the immunological interference and antimicrobial nature of MmLec which would be supportive in establishing a potential therapeutic tool and to develop better and novel disease control strategies in shrimp and farmed aquaculture industries as well as in health management.

Anti-cancer, anti-biofilm, and anti-inflammatory properties of hen's albumen: A photodynamic approach.

The albumen plays a major role in the protection of eggs against microorganisms. It contains an arsenal of natural antimicrobial molecules and antibacterial proteins, including the well-known ovotransferrin and lysozyme, which exert their activities against a range of bacteria. In the present study, the hen's albumen extract treated with the dried insect body of blister beetle M. pustulata was assessed for antibacterial, antibiofilm, anti-inflammatory and anti-proliferative activity. The zone of inhibition against Gram positive E. faecalis and S. aureus was 10.8 mm and 12.1 mm respectively at 100 µg mL-1. However, it was 13.6 mm and 15.3 mm for Gram negative P. aeruginosa and P. vulgaris respectively. The biofilm of tested bacteria was significantly inhibited at 100 µg mL-1. The hydrophobicity of bacterial biofilms was considerably condensed after treatment with the hen's albumen extracts at 100 µg mL-1. The anti-inflammatory activity of hen's albumen extracts was confirmed by the inhibition of cyclooxygenase (COX) enzyme to 84.91% at 100 µg mL-1 with the relative IC50 of 8.26 µg mL-1. The albumen extract effectively inhibited the viability (23.61%) of HepG2 hepatic cancer cells at 100 µg mL-1. The anti-proliferative activity of the albumen extracts was further revealed by the induction of HepG2 apoptotic cell morphology. This study concludes that the hen's albumen extract treated with M. pustulata is a natural therapeutic agent to treat biofilm associated clinical bacteria, inflammations and human hepatic cancer cells.

Garlic clove extract assisted silver nanoparticle - Antibacterial, antibiofilm, antihelminthic, anti-inflammatory, anticancer and ecotoxicity assessment.

Facile and low cost garlic clove extract based silver nanoparticles was synthesized and its broad spectrum of therapeutic activity including antibiofilm, antiparasitic and anti-breast cancer activity was evaluated. The synthesized garlic­silver nanoparticles (G-AgNPs) were characterized by various physico-chemical techniques. G-AgNPs showed good optical property, highly crystalline nature, spherical shape and uniformly dispersed with size measuring between 10 and 50 nm. G-AgNPs have shown greater anti-bacterial and antibiofilm activity on clinically important pathogens methicillin-resistant S. aureus and P. aerigunosa at 100 µg ml-1. The efficacy of G-AgNPs against earthworm evidenced its effectiveness as anti-helminthic agent in treating intestinal parasites. The significant inhibition of BSA protein denaturation proves its anti-inflammatory property. In addition, G-AgNPs have shown remarkable anticancer effect and significantly inhibited the human breast cancer cell (MCF-7) viability at 100 µg ml-1 after 24 h. A noticeable change in the morphology of MCF-7 cells was also noticed. G-AgNPs were non-toxic to human HEK293 embryonic cells. Also, the non-toxic nature of G-AgNPs to C. cornuta and no morphological, physiological changes proved its safety to the environment. It is concluded that G-AgNPs have a broad range of biological applications and it can be used as an eco-friendly material without having negative effects in the environment.

Antimicrobial and biochemical characterization of a C-type lectin isolated from pearl spot (Etroplus suratensis).

The present study reveals purification and characterization of a C-type lectin from the serum of pearl spot, Etroplus suratensis (Es-Lec). The Es-Lec was purified by affinity chromatography with mannose coupled sepharose CL-4B column and it exhibits single band with a molecular weight of 75 kDa in SDS-PAGE. The surface morphology of purified Es-Lec displays the homogeneous nature of protein. A distinct peak with a retention time of 2.958 min was appeared in high performance liquid chromatography (HPLC), X-ray diffraction (XRD) analysis expresses a single peak at 31.8372̊ and MALDI-TOF peaks which shows the purity and crystalline nature of the protein respectively. Functional analysis of purified Es-Lec exhibits yeast agglutination activity against Saccharomyces cerevisiae and has the ability to agglutinate the human erythrocytes, which was observed by light microscopy and haemagglutination inhibition was also done. In addition, purified Es-Lec showed the broad spectrum of antibacterial activity against Gram negative Vibrio parahaemolyticus and Aeromonas hydrophila. Antibiofilm potential of purified Es-Lec against selected Gram-negative bacteria exhibited the disruption of biofilm architecture at the concentration of 50 µg ml-1 and also it exhibited antiviral and anticancer activity.

Crustin-capped selenium nanowires against microbial pathogens and Japanese encephalitis mosquito vectors - Insights on their toxicity and internalization.

Herein, we reported a method to synthesize selenium nanowires (Cr-SeNWs) relying to purified cysteine-rich antimicrobial peptide crustin in presence of ascorbic acid. Cr-SeNWs were characterized by UV-vis, XRD, FTIR and Raman spectroscopy, as well as SEM, HR-TEM and EDAX. The UV-vis spectroscopy peak was noted at 350 nm. XRD showed the crystalline nature of Cr-SeNWs through diffraction peaks observed 2θ at 12° and 28° corresponding to (020), and (241) lattice planes, respectively. HR-TEM results shed light on the size of Cr-SeNWs, ranging from 17 to 47 nm. Raman spectroscopy and EDAX analysis of Cr-SeNWs showed presence of 57% selenium element. Furthermore, Cr-SeNWs showed higher antimicrobial activity on Gram-positive bacteria (Staphylococcus aureus, Enterococcus faecalis) over Gram-negative ones (Pseudomonas aeruginosa, Escherichia coli). The zone of inhibition was larger on S. aureus (50 µg/ml = 4.0 mm, 75 µg/ml = 7.2 mm) and E. faecalis (50 µg/ml = 3.1 mm, 75 µg/ml = 5.1 mm), over P. aeruginosa (50 µg/ml = 2.1 mm, 75 µg/ml = 4.8 mm), E. coli (50 µg/ml = 1.3 mm, 75 µg/ml = 4.3 mm) bacteria. The antibiofilm activity of Cr-SeNWs was also investigated and biofilm reduction was observed at 75 µg/ml. In addition, Cr-SeNWs were highly effective as larvicides against Zika virus and Japanese encephalitis mosquito vectors, i.e., Culex quinquefasciatus and Culex tritaeniorhynchus, with LC50 values of 4.15 and 4.85 mg/l, respectively. The nanowire toxicity and internalization was investigated through confocal laser scanning microscopy and histological studies. To investigate the potential of Cr-SeNWs for real-world applications, we also evaluated Cr-SeNWs in hemolytic assays, showing no cytotoxicity till 5 mg/ml. Besides, higher antioxidant activity at the concentration at 100 µg/ml was noted, if compared with purified crustin. The strong antioxidant potential of this nanomaterial can be helpful to boost the shelf-life potential of Cr-SeNWs-based pesticides and antimicrobials.

Bioaccumulation, cytotoxicity and oxidative stress of the acute exposure selenium in Oreochromis mossambicus.

Selenium (Se) is an essential trace-element that becomes toxic when present at high concentrations for aquatic organisms. The knowledge about the mechanism of Se toxicity in freshwater ecosystem is still poorly studied. Thus the aim of the present study was to assess the impact of environmentally relevant concentrations of Se toxicity: 5, 10, 25, 50 and 100 µg/L or water only (control) for periods of 96 hour (h) to test for Se accumulation (gill, liver and brain), its effects on enzymatic and non-enzymatic antioxidant defenses (gill and liver), oxidative stress effects on lipid, protein (gill and liver), DNA (liver) and inhibition of AchE (brain) activity were measured in Mozambique tilapia, Oreochromis mossambicus. Our result showed that Se accumulation was observed in the gill, liver and brain tissues of fish exposed to different concentrations and accumulation varied upon different tissues. Enzymatic (SOD, CAT, GPx and GST) and non-enzymatic (GSH and MT) antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-s-transferase (GST) were significantly increased after 96 h exposure of higher concentrations Se in the gill and liver tissue with the exception of GST activity was significantly inhibited in liver after 96 h exposure of higher concentrations of Se. In contrast, catalase (CAT) activities were inhibited for both tissues of Se exposure at 96 h. Reduced glutathione (GSH) and Metallothionein (MT) levels were increased in the gill and liver tissues after exposure to Se for 96 h. We also observed that Se affected antioxidant defense, increasing oxidative stress indicator of lipid peroxidation (LPO) and protein carbonyl (PCO) in gill and liver tissues of fish exposed to Se for 96 h at the concentration dependent manner. Increased DNA damage scores observed in liver tissue of fish exposed to Se for concentrations dependent manner, indicating potential of Se on fish. We also observed inhibition of acetylcholine esterase (AchE) activity in brain tissue of fish exposed to Se for higher concentrations. The changes in these parameters can be used as suitable biomarkers for monitoring the toxicity of Se in the aquatic environment.

Investigation of antioxidant and anticancer potential of fucoidan from Sargassum polycystum.

The present study was aimed to evaluate the antioxidant and anticancer potential of fucoidan isolated from Sargassum polycystum. The isolated fucoidan was successfully purified by DEAE cellulose-ion exchange chromatography and dialysis. Totally four active fractions (F1-F4) were collected and explored its chemical constitution by calorimetric assays. Among them, fraction 2 (F2) showed the higher yield percentage, fucose and sulphate content. Further, monosaccharide composition, structural and functional properties of the F2 was analyzed by HPLC, FTIR and NMR. F2 shows highest DPPH radical scavenging activity (55.94 ±â€¯0.69%), reducing power (0.33 absorbance rate), hydrogen peroxide scavenging activity (71.76 ±â€¯2.14%) and nitric oxide radical scavenging activity (51.81 ±â€¯1.04%) at 1000 µg/ml. The cell viability of MCF-7 and HCT-15 cell lines was proportionate to the concentration of F2 with an estimated IC50 was 20 and 50 µg/ml respectively. The fluorescence and confocal laser scanning microscopic analysis demonstrated the apoptotic morphological changes and cell mediated death in F2 treated cancer cells. Higher amount of LDH release was found in the F2 treated cancer cells than the control group. Thus, the present finding proved that the isolated F2 encompasses significant antioxidant and anticancer property.

Identification, characterization and immune response of prophenoloxidase from the blue swimmer crab Portunus pelagicus and its antibiofilm activity.

Prophenoloxidase is a conserved Cu-containing enzyme acting as a major defense molecule in the immune response of crustaceans. In the present research, we purified prophenoloxidase from the haemolymph of Portunus pelagicus (Pp-proPO) by Blue Sepharose CL-6B chromatography. Pp-proPO exhibited only one band with molecular weight of 75kDa on SDS-PAGE. The purified Pp-proPO was characterized through X-ray diffraction (XRD) and high-performance liquid chromatography (HPLC). Pp-proPO showed phagocytic activity on the yeast Saccharomyces cerevisiae as well as encapsulation on sepharose CL-6B beads associated with CM sepharose and beads of sodium alginate. Pp-proPO also led to strong agglutination on human erythrocytes. Furthermore, Pp-proPO showed magnified PO activity when altered with activated particles acting as pathogen combined molecular patterns (PAMPs), metal ions or other chemicals. Pp-proPO showed relevant antibiofilm activity on Gram negative bacteria Pseudomonas aeruginosa and Escherichia coli. Overall, the above results allowed us to claim that Pp-proPO play a key role in immune defense mechanisms of P. pelagicus crabs, in particular towards microbial pathogens; notably we added basic information to the functional characterization of Pp-proPO, as well as to understand its immunological role in crustaceans defense systems.

Proteomics analysis of crude squid ink isolated from Sepia esculenta for their antimicrobial, antibiofilm and cytotoxic properties.

The present study deals with the proteomics analysis of crude squid ink isolated from Sepia esculenta for their antibacterial, antifungal, antibiofilm and cytotoxic properties. To achieve this, SDS-PAGE was used to separate proteins as bands, In-gel trypsin digested and analyzed by MALDI-TOF mass spectrometry. A total of 4 bands were identified by MASCOT search analysis namely astacin-like squid metalloprotease type I (ASMT-I), 70 kDa neurofilament protein (NP), uncharacterized protein LOC106181966 isoform X1 (UP-Iso-X1) and Ommochrome-binding protein (Oc-BP). Further, the obtained crude squid proteins were subjected to antimicrobial and antibiofilm activities against pathogenic bacterial and fungal strains respectively. Further, MTT assay was also carried out to deliberately explain the cytotoxic ability of crude squid ink protein against MCF-7 breast cancer cell lines. The results from the study revealed that, the proteins are shown to be toxic against pathogenic strains and breast cancer cell lines in a dose-dependent manner. More importantly, the proteins are well enough to eradicate biofilms substantiated by light and confocal laser scanning microscopic observations. Altogether, the crude squid ink proteins hampered the growth of breast cancer cells with an IC50 value of 65.3 ±â€¯0.46 µg mL-1. In conclusion, it is believed that the proteins from crude squid ink will provide new insights in hampering bacterial biofilms and cancer in near future.

High efficacy of (Z)-γ-bisabolene from the essential oil of Galinsoga parviflora (Asteraceae) as larvicide and oviposition deterrent against six mosquito vectors.

The eco-friendly management of mosquitoes with novel and effective larvicides and oviposition deterrents is a crucial challenge to prevent outbreaks of mosquito-borne diseases. However, most of the herbal formulations tested in these years showed LC50 values higher of 40 ppm, and significant oviposition deterrent activity only when tested at relatively higher doses (> 50 µg/ml). Herein, we studied the chemical composition of the Galinsoga parviflora essential oil (EO). This plant is an annual herb native to South America naturalized all over the world. We tested the EO larvicidal and oviposition deterrent action on 6 mosquito species. Totally 37 compounds were identified in the EO of G. parviflora by GC and GC-MS analyses. The major constituent was (Z)-γ-bisabolene (38.9%). The G. parviflora EO and (Z)-γ-bisabolene showed acute toxicity on An. stephensi (LC50 = 31.04 and 2.04 µg/ml, respectively), Ae. aegypti (LC50 = 34.22 and 2.26 µg/ml, respectively), Cx. quinquefasciatus (LC50 = 37.10 and 2.47 µg/ml, respectively), An. subpictus (LC50 = 40.97 and 4.09 µg/ml, respectively), Ae. albopictus (LC50 = 45.55 and 4.50 µg/ml, respectively) and Cx. tritaeniorhynchus (LC50 = 49.56 and 4.87 µg/ml, respectively) larvae. Furthermore, the oviposition deterrent potential of the G. parviflora EO and (Z)-γ-bisabolene was studied on six mosquito vectors, showing that 25 µg/ml of (Z)-γ-bisabolene led to an Oviposition Activity Index lower of - 0.79 in all tested mosquito vectors. Overall, all larvicidal LC50 values estimated for (Z)-γ-bisabolene were lower than 5 µg/ml. This result far encompasses current evidences of toxicity reported for the large majority of botanical products currently tested against mosquito young instars, allowing us to propose this compound as an highly effective mosquito larvicide and oviposition deterrent.

Dietary supplementation of probiotic Bacillus licheniformis Dahb1 improves growth performance, mucus and serum immune parameters, antioxidant enzyme activity as well as resistance against Aeromonas hydrophila in tilapia Oreochromis mossambicus.

The present study evaluated the dietary supplementation of probiotic Bacillus licheniformis Dahb1 on the growth performance, immune parameters and antioxidant enzymes activities in serum and mucus as well as resistance against Aeromonas hydrophila in Mozambique tilapia Oreochromis mossambicus. Fish (24 ±â€¯2.5 g) were fed separately with three diets, 1) commercial diet (control), 2) diet containing probiotic at 105 cfu g-1 (D1) and 3) diet containing probiotic at 107 cfu g-1 (D2) for 4 weeks. Growth performance in term of final weight (FW) specific growth rate (SGR) and feed conversion ratio (FCR), immune parameters of total protein (TP), alkaline phosphatase (ALP), myeloperoxidase (MPO), lysozyme (LYZ), reactive oxygen species (ROS), reactive nitrogen species (RNS) and antioxidant parameters of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in serum and mucus were evaluated after 2nd and 4th weeks. The FW, SGR, and FCR of fish fed with D1 and D2 significantly improved (p < 0.05). The activities of ALP, LYZ and MPO in the mucus were significantly higher (p < 0.05) in fish that fed D1 and D2. The TP, ROS, RNS, SOD and GPx in the serum were significantly higher (p < 0.05) in fish that fed D1 and D2. In addition, the challenge test showed that fish fed D1 and D2 enhanced significantly (p < 0.05) the resistance against A. hydrophila (1 × 107 cells ml-1). In conclusion, probiotic B. licheniformis Dahb1 can be applied in diet at 107 cfu g-1 to improve healthy status and resistance against A. hydrophila in tilapia farming.