Development of Paper-Based Fluorescent Molecularly Imprinted Polymer Sensor for Rapid Detection of Lumpy Skin Disease Virus.

Lumpy Skin Disease (LSD) is a notifiable viral disease caused by Lumpy Skin Disease virus (LSDV). It is usually associated with high economic losses, including a loss of productivity, infertility, and death. LSDV shares genetic and antigenic similarities with Sheep pox virus (SPV) and Goat pox (GPV) virus. Hence, the LSDV traditional diagnostic tools faced many limitations regarding sensitivity, specificity, and cross-reactivity. Herein, we fabricated a paper-based turn-on fluorescent Molecularly Imprinted Polymer (MIP) sensor for the rapid detection of LSDV. The LSDV-MIPs sensor showed strong fluorescent intensity signal enhancement in response to the presence of the virus within minutes. Our sensor showed a limit of detection of 101 log10 TCID50/mL. Moreover, it showed significantly higher specificity to LSDV relative to other viruses, especially SPV. To our knowledge, this is the first record of a paper-based rapid detection test for LSDV depending on fluorescent turn-on behavior.

Efficacy of zinc oxide and copper oxide nanoparticles on virulence genes of avian pathogenic E. coli (APEC) in broilers.

BACKGROUND: Colibacillosis is one of the broilers' most dominant bacterial diseases, either as a primary or a secondary infection. As E. coli antimicrobial drug resistance is rising; there is a need to develop new approaches to its control. In light of this, a comparative study of the in-vitro antibacterial activity of Arabic gum stabilized zinc and copper nanoparticles (AG-ZnNPs and AG-CuNPs) against PCR-identified field avian pathogenic E. coli (APEC) strains and virulence genes (ibeA, hlyA, iss, pap C and ompA) was applied to study the therapeutic effect of zinc and copper nanoparticles to be used as an antibiotic alternative (Nanobiotic). Furthermore, the in-vivo effects of CuNPs were evaluated. Additionally, the CuNPs liver and muscle residues with or without infection were examined. The eighty broilers were divided into four groups; G1: negative control, G2: infected control with E. coli O17, G3: non-infected treated (AG-CuNPs 50 mg/kg body weight), and G4: infected treated (AG-CuNPs 50 mg/kg body weight). AG-CuNPs treatment was given to broilers for five days in drinking water. RESULTS: E. coli was isolated from diseased broilers at an average incidence rate of 20% from intestinal and liver samples. All identified serotypes (O17, O78, O91, O121, and O159) were resistant to AG-ZnNPs and sensitive to AG-CuNPs. AG-CuNPs minimal inhibitory and bactericidal concentrations (MIC and MBC) for O17 were 7.5 and 60 mg/ml, respectively. Conventional uniplex PCR results showed that strain O17 contained virulence genes (ibeA, hlyA, iss, and papC), where AG-CuNPs significantly reduced the expression of all target genes when examined by Real-time quantitative PCR. Additionally, the bactericidal activity of AG-CuNPs on O17 was 100% at 20 minutes and 40 mg/ml and confirmed by transmission electron microscopy. Furthermore, no mortality was recorded in treated groups compared to G2. Subsequently, no E. coli was re-isolated from the liver in the G4 after treatment. The total protein, albumin, globulin, and lysozyme activity were significantly increased in G4 compared to G2, while the activities of liver enzymes (alanine aminotransferase (ALT), Gamma-glutamyl transferase (GGT), and alkaline phosphatase (ALP)) were markedly decreased in G4 compared to G2. Additionally, uric acid, creatinine, and C-reactive protein levels were decreased in G4 compared to G2. However, the liver enzymes, kidney functions, C-reactive protein levels, and Cu residues were non-significantly changed in G4 compared to G1. CONCLUSION: Green synthesized AG-CuNPs are recommended as an effective antimicrobial alternative against APEC strains.

Assessing the In Vivo Biocompatibility of Molecularly Imprinted Polymer Nanoparticles.

Molecularly imprinted polymer nanoparticles (nanoMIPs) are high affinity synthetic receptors which show promise as imaging and therapeutic agents. Comprehensive analysis of the in vivo behaviour of nanoMIPs must be performed before they can be considered for clinical applications. This work reports the solid-phase synthesis of nanoMIPs and an investigation of their biodistribution, clearance and cytotoxicity in a rat model following both intravenous and oral administration. These nanoMIPs were found in each harvested tissue type, including brain tissue, implying their ability to cross the blood-brain barrier. The nanoMIPs were cleared from the body via both faeces and urine. Furthermore, we describe an immunogenicity study in mice, demonstrating that nanoMIPs specific for a cell surface protein showed moderate adjuvant properties, whilst those imprinted for a scrambled peptide showed no such behaviour. Given their ability to access all tissue types and their relatively low cytotoxicity, these results pave the way for in vivo applications of nanoMIPs.

In vivo study of dose-dependent antioxidant efficacy of functionalized core-shell yttrium oxide nanoparticles.

Herein, we assess the dose-dependent antioxidant efficacy of ultrafine spherical functionalized core-shell yttrium oxide nanoparticles (YNPs) with a mean size of 7-8 nm and modified with poly EGMP (ethylene glycol methacrylate phosphate) and N-Fluorescein Acrylamide. The antioxidant properties of these nanoparticles were investigated in three groups of Sprague-Dawley rats (10 per group) exposed to environmental stress daily for 1 week and one control group. Groups 2 and 3 were intravenously injected twice a week with YNPs at 0.3 and 0.5 mg at 2nd and 5th day of environmental stress exposure respectively. Different samples of blood and serum were collected from all experimental groups at end of the experiment to measure oxidative biomarkers such as total antioxidant capacity (TAC), hydroxyl radical antioxidant capacity (HORAC), oxygen radical antioxidant capacity (ORAC), malondialdehyde (MDA), and oxidants concentration as hydrogen peroxide (H2O2). The liver, brain, and spleen tissues were collected for fluorescence imaging and histopathological examination in addition to brain tissue examination by transmission electron microscope (TEM). Inductively coupled plasma-mass spectrometry (ICP-MS) was used to estimate YNPs translocation and concentration in tissues which is consecutively dependent on the dose of administration. Depending on all results, poly EGMP YNPs (poly EGMP yttrium oxide nanoparticles) can act as a potent direct antioxidant in a dose-dependent manner with good permeability through blood-brain barrier (BBB). Also, the neuroprotective effect of YNPs opening the door to a new therapeutic approach for modulating oxidative stress-related neural disorders. HIGHLIGHTS: • The dose-dependent antioxidant efficacy of ultrafine spherical functionalized core-shell yttrium oxide nanoparticles (YNPs) with a mean size of 7-8 nm and modified with poly EGMP (ethylene glycol methacrylate phosphate) and N-Fluorescein Acrylamide was assessed. • The dose of administration directly affecting the brain, liver, and spleen tissues distribution, retention, and uptake of YNPs and direct correlation between the absorbed amount and higher dose administered. • YNPs can act as a potent direct antioxidant in a dose-dependent manner with good permeability through blood-brain barrier (BBB).

Assessment the efficacy of some various treatment methods, in vitro and in vivo, against Aeromonas hydrophila infection in fish with regard to side effects and residues.

Aeromonas hydrophila is an opportunistic bacteria with an overwhelming impact on fish farming industry especially with upraising of drug resistant mutants. This study aimed to evaluate and compare the therapeutic and side effects of levofloxacin (LEV), chitosan-nanoparticles (CNPs), and fructooligosaccharides (FOS) in control of this infection in tilapia. A total of 160 Nile-tilapia divided into 8-groups; G1: negative-control, G2: infected-control, G3: non-infected-(levofloxacin (LEV) 10 mg/kg bwt), G4: non-infected-(chitosan-nanoparticles (CNPs) 1 g/kg ration), G5: non-infected-(fructooligosaccharides (FOS) 20 g/kg ration), G6: infected-LEV, G7: infected-CNPs and G8: infected-FOS for 7 days. MICs were (0.125 µg/ml and 1.25 mg/ml) for LEV and CNPs respectively. No mortalities or significant adverse effects were recorded in non-infected treated-groups while infected were (20%) LEV, (30%) CNPs, (40%) FOS and (70%) G2. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) decreased by LEV and CNPs than FOS while all increased total protein (TP) and albumin than G2. Malondialdehyde (MDA) significantly decreased and superoxide dismutase (SOD) and reduced glutathione (GSH) increased in all infected-treated groups than G2 in various degrees. Urea and creatinine descending order were FOS, LEV then CNPs decreased significantly than G2. LEV musculature residues, using HPLC, decreased gradually till the 5th day; 621.00 ± 0.66, 270.00 ± 0.48 then 64.00 ± 0.40, and 471.00 ± 0.79, 175.00 ± 0.52 ppb then not detected at 1st, 3rd, and 5th days of withdrawal in non-infected and infected groups respectively. Finally, LEV and CNPs were superior as bactericidal, decreasing mortalities and enzyme activities while CNPs and FOS increased performance, non-specific immunity, and antioxidant biomarkers.

Functionalized Core-Shell Yttrium Oxide Nanoparticles as Antioxidants Agents in Heat Stressed Rats.

Herein, we assess the antioxidant potential of core-shell yttrium oxide nanoparticles functionalized with ethylene glycol methacrylate phosphate (EGMP). The antioxidant properties of these nanoparticles were investigated in heat stressed models of 21 rats (heat stressed group, group B). Different samples of blood, serum, and tissue homogenate were collected at different time intervals in order to measure oxidative biomarkers such as enzymatic antioxidants (SODs, GPX, GST, GR, and TAC) and oxidative byproducts (MDA, PC, and 8-OHdG). Liver specimens of prophylactic group and heat stressed ones were also histopathologically examined 2 h post NPs injection. The measurements of oxidative biomarkers were complementary with histopathological findings and confirmed the antioxidant properties of poly EGMP yttrium oxide NPs.

Biocompatibility and biodistribution of surface-modified yttrium oxide nanoparticles for potential theranostic applications.

The surface of ultrafine yttrium oxide nanoparticles (NPs) with mean size of 7-8 nm was modified with a functional polymer layer to improve their dispersion and impart fluorescent properties for imaging purposes. Surface functionalization was achieved by silanization of yttrium oxide NPs with 3-trimethoxysilylpropyl methacrylate followed by grafting of a co-polymer made of acrylic acid (AA) and ethylene glycol methacrylate phosphate (EGMP). The polymer shell decreases the surface energy of NPs, enhances their polarity, and, as a result, improves their colloidal stability. The synthesized NPs are capable of scavenging free radicals and for this reason have therapeutic potential that warrants further investigations. Furthermore, these stabilized core-shell NPs showed a very low cytotoxicity, confirming that the polymer shell sensibly improves the biocompatibility of bare yttrium oxide NPs, which are otherwise toxic on their own. Poly-EGMP yttrium NPs proved to be safe up to 0.1 mg/g body weight in 1 month old Sprague-Dawley rats, showing also the ability to cross the blood-brain barrier short time after tail injection. The surface modification of yttrium NPs here described allows these NPs to be potentially used in theranostics to reduce neurodegenerative damage due to the heat stress.