Search results for: antimicrobial peptide

Authors: Tesfay Gebremichael Reda

Abstract:

Nanoparticle technology is fast progressing and is being employed in innumerable medical applications. At this time, the public's health is seriously threatened by the rise of bacterial strains resistant to several medications. Metal nanoparticles are a potential alternate approach for tackling this global concern, and this is the main focus of this study. The citrate precursor sol-gel synthesis method was used to synthesize the, Niₓ Co(₁-ₓ) Fe₂ O₄, (where x = 0.0:0.2:1.0) nanoparticle. XRD identified the development of the cubic crystal structure to have a preferential orientation along (311), and the average particle size was found to be 29-38 nm. The average crystallizes assessed with ImageJ software and origin 22 of the SEM are nearly identical to the XRD results. In the created NCF NPs, the FT-IR spectroscopy reveals structural examinations and the redistribution of cations between octahedral (505-428 cm-1) and tetrahedral (653-603 cm-1) locales. Finally, the decrease of coercive fields HC, 2384 Oe to 241.93 Oe replacement of Co²+ cation with Ni²+. Band gap energy rises as Ni concentration increases, which may be attributed to the fact that the ionic radii of Ni²+ ions are smaller than that of Co²+ ions, which results in a strong electrostatic interaction. On the contrary, except at x = 0.4, the dielectric constant decreases as the nickel concentration increases. According to the findings of this research work, nanoparticles composed of Ni₀.₄ Co₀.₄ Fe₂ O₄ have demonstrated a promising value against S. aureus and E. coli, and it suggests a proposed model for their potential use as a new source of antibacterial agents.

Keywords: antimicrobial, band gap, citrate precursor, dielectric, nanoparticle

Procedia PDF Downloads 28

Authors: Sanaa Bardaweel

Abstract:

Inorganic polyphosphate (poly P) is present in all living forms tested to date, from each of the three kingdoms of life. Studied mainly in prokaryotes, poly P and its associated enzymes are vital in diverse basic metabolism, in at least some structural functions and, notably, in stress responses. These plentiful and unrelated roles for poly P are probably the consequence of its presence in life-forms early in evolution. The genomes of many bacterial species, including pathogens, encode a homologue of a major poly P synthetic enzyme, poly P kinase 1 (PPK1). Genetic deletion of ppk1 results in reduced poly P levels and loss of pathogens virulence towards protozoa and animals. Thus far, no PPK1 homologue has been identified in higher-order eukaryotes and, therefore, PPK1 represents a novel target for chemotherapy. The idea of the current study is to purify the PPK1 from Escherichia coli to homogeneity in order to study the effect of active site point mutations on PPK1 catalysis via the application of site-directed mutagenesis strategy. The knowledge obtained about the active site of PPK1 will be utilized to characterize the catalytic and kinetic mechanism of PPK1 with model substrates. Comprehensive understanding of the enzyme kinetic mechanism and catalysis will be used to design and screen a library of synthetic compounds for potential discovery of selective PPK1-inhibitors.

Keywords: antimicobial, Escherichia coli, inorganic polyphosphate, PPK1-inhibitors

Procedia PDF Downloads 279

Authors: A. A. Dehpour, B. Eslami, S. Rezaie, S. F. Hashemian, F. Shafie, M. Kiaie

Abstract:

The aims of study were investigation on chemical composition essential oil and the effect of extract of Coronilla varia on antimicrobial and cytotoxicity activity. The essential oils of Coronilla varia is obtained by hydrodistillation and analyzed by (GC/MS) for determining their chemical composition and identification of their components. Antibacterial activity of plant extract was determined by disc diffusion method. The effect of hydroalcolic extracts from Cornilla varia investigated on MCF7 cancer cell line by MTT assay. The major components were Caryophyllene Oxide (60.19%), Alphacadinol (4.13%) and Homoadantaneca Robexylic Acid (3.31%). The extracts from Coronilla varia had interesting activity against Proteus mirabilis in the concentration of 700 µg/disc and did not show any activity against Staphylococus aureus, Bacillus subtillis, Klebsiella pneumonia and Entrobacter cloacae. The positive control, Ampicillin, Chloramphenicol and Cenphalothin had shown zone of inhibition resistant all bacteria. Corohilla varia ethanol extract could inhibit the proliferation of MCF7 cell line in RPMI 1640 medium. IC50 5(mg/ml) was the optimum concentration of extract from Coronilla varia inhibition of cell line growth. The MCF7 cancer cell line and Proteus mirabilis were more sensitive to Coronilla varia ethanol extract.

Keywords: Coronilla varia, essential oil, antibacterial, anticancer, hela cell line

Procedia PDF Downloads 390

Authors: Sanja O. Podunavac-Kuzmanović, Lidija R. Jevrić, Strahinja Z. Kovačević

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In last decade, metal-organic complexes have captured intensive attention because of their wide range of biological activities such as antibacterial, antifungal, anticancerous, antimicrobial and antiHIV. Therefore, it is of great importance for the development of coordination chemistry to explore the assembly of functional organic ligands with metal ion and to investigate the relationship between the structure and property. In view of our studies, we reasoned that benzimidazoles complexed to metal ions could act as a potent antibacterial agents. Thus, we have bioassayed the inhibitory potency of benzimidazoles and their metal salts (Co or Ni) against Gram negative bacteria Escherichia coli. In order to validate our in vitro study, we performed in silico studies using molecular docking software’s. The investigated compounds and their metal complexes (Co, Ni) showed good antibacterial activity against Escherichia coli. In silico docking studies of the synthesized compounds suggested that complexed benzimidazoles have a greater binding affinity and enhanced antibacterial activity in comparison with noncomplexed ligands. In view of their enhanced inhibitory properties we propose that the studied complexes can be used as potential pharmaceuticals. This study is financially supported by COST action CM1306 and the project No. 114-451-347/2015-02, financially supported by the Provincial Secretariat for Science and Technological Development of Vojvodina.

Keywords: benzimidazoles, complexes, antibacterial, Escherichia coli, metal

Procedia PDF Downloads 317

Authors: Rongzhou Wang

Abstract:

Research on designing nano-antibacterial agent with potent and long-lasting antibacterial property is demanding and provoking work. In this study, a core-shell AgBr/cationic polymer nanocomposite (AgBr/NPVP-H10) were synthesized and its structure confirmed by Fourier Transform Infrared Spectrometer (FT-IR), Nuclear Magnetic Resonance (1H NMR) and X-ray diffraction (XRD), and the cationic polymer contents were determined with Thermal Gravimetric Analyzer (TGA). The morphology was directly observed by Transmission Electron Microscope (TEM) which showed that the nanoparticle contains single core and organic shell and had an average diameter of 30.1 nm. The antibacterial test against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli illuminated that this nanocomposite had potent bactericidal activity, which can be attributed to the contact-killing of cationic polymers and releasing-killing of Ag+ ions. In addition, cationic polymer encapsulating AgBr cores gave the resin discs sustained release of Ag+ ions, which may result in long-lasting bactericidal activity. The AgBr/NPVP-H10 nanoparticle with the dual bactericidal capability and long term antimicrobial effect is a promising material aimed at preventing bacterial infection.

Keywords: core-shell nanocomposite, cationic polymer, dual antibacterial capability, long-lasting antibacterial activity

Procedia PDF Downloads 191

Authors: Zohra Benfodda, Valentin Duvauchelle, Chaimae Majdi, David Bénimélis, Catherine Dunyach-Remy, Patrick Meffre

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New antibiotics are necessary to treat microbial pathogens, especially ESKAPE pathogens that are becoming increasingly resistant to available treatment. Despite the medical need, the number of newly approved drugs continues to decline. The majority of antibiotics under clinical development are natural products or derivatives thereof. 43 juglone/naphthazarin derivatives were synthesized using Minisci-type direct C–H alkylation and evaluated for their antibacterial properties against various clinical and reference Gram-positive MSSA, clinical Gram-positive MRSA. Different compounds of the synthesized series showed promising activity against clinical and reference MSSA (MIC: 1–8 μg/ml) and good efficacy against clinical MRSA (MIC: 2–8 μg/ml) strains. The synergistic effects of active compounds were evaluated with reference antibiotics (vancomycin and cloxacillin), and it was found that the antibiotic combination with those active compounds efficiently enhanced the antimicrobial activity and consequently the MIC values of reference antibiotics were lowered up to 1/16th of the original MIC. These synthesized compounds did not present hemolytic activity on sheep red blood cells. In addition to the in silico prediction of ADME profile parameter which is promising and encouraging for further development.

Keywords: juglone, naphthazarin, antibacterial, clinical MRSA, synergistic studies, MIC determination

Procedia PDF Downloads 126

Authors: Evgeniya Y. Yuzbasheva, Tigran V. Yuzbashev, Natalia I. Perkovskaya, Elizaveta B. Mostova

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Cell-surface display of lipase is of great interest as it has many applications in the field of biotechnology owing to its unique advantages: simplified product purification, and cost-effective downstream processing. One promising area of application for whole-cell biocatalysts with surface displayed lipase is biodiesel synthesis. Biodiesel is biodegradable, renewable, and nontoxic alternative fuel for diesel engines. Although the alkaline catalysis method has been widely used for biodiesel production, it has a number of limitations, such as rigorous feedstock specifications, complicated downstream processes, including removal of inorganic salts from the product, recovery of the salt-containing by-product glycerol, and treatment of alkaline wastewater. Enzymatic synthesis of biodiesel can overcome these drawbacks. In this study, Lip2p lipase was displayed on Yarrowia lipolytica cells via C- and N-terminal fusion variant. The active site of lipase is located near the C-terminus, therefore to prevent the activity loosing the insertion of glycine-serine linker between Lip2p and C-domains was performed. The hydrolytic activity of the displayed lipase reached 12,000–18,000 U/g of dry weight. However, leakage of enzyme from the cell wall was observed. In case of C-terminal fusion variant, the leakage was occurred due to the proteolytic cleavage within the linker peptide. In case of N-terminal fusion variant, the leaking enzyme was presented as three proteins, one of which corresponded to the whole hybrid protein. The calculated number of recombinant enzyme displayed on the cell surface is approximately 6–9 × 105 molecules per cell, which is close to the theoretical maximum (2 × 106 molecules/cell). Thus, we attribute the enzyme leakage to the limited space available on the cell surface. Nevertheless, cell-bound lipase exhibited greater stability to short-term and long-term temperature treatment than the native enzyme. It retained 74% of original activity at 60°C for 5 min of incubation, and 83% of original activity after incubation at 50°C during 5 h. Cell-bound lipase had also higher stability in organic solvents and detergents. The developed whole-cell biocatalyst was used for recycling biodiesel synthesis. Two repeated cycles of methanolysis yielded 84.1–% and 71.0–% methyl esters after 33–h and 45–h reactions, respectively.

Keywords: biodiesel, cell-surface display, lipase, whole-cell biocatalyst

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Authors: Farzad Doostishoar, Abbas Pardakhty, Abdolreza Hassanzadeh, Sudeh salarpour, Elham Sharif

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Background: Sericin is a protein extracted from silk and has antioxidant, antimicrobial, antineoplastic, wound healing and moisturizing properties. Different cosmetic formulation of sericin is available in different countries such as Japan and the other south-eastern Asian countries. We formulated and evaluated the sunscreen properties of topical formulations of sericin by an in vitro method. Method: Niosomes composed of sorbitan palmitate (Span 40), polysorbate 40 (Tween 40) and cholesterol (300 µmol, 3.5:3.5:3 molar ratio) were prepared by film hydration technique. Sericin was dissolved in normal saline and the lipid hydration was carried out at 60°C and the niosomes were incorporated in a Carbomer gel base. A W/O cream was also prepared and the release of sericin was evaluated by using Franz diffusion cell. Particle size analysis, sericin encapsulation efficiency measurement, morphological studies and stability evaluation were done in niosomal formulations. SPF was calculated by using Transpore tape in vitro method for both formulations. Results: Niosomes had high stability during 6 months storage at 4-8°C. The mean volume diameter of niosomes was less than 7 µm which is ideal for sustained release of drugs in topical formulations. The SPF of niosomal gel was 25 and higher than sericin cream with a diffusion based release pattern of active material. Conclusion: Sericin can be successfully entrapped in niosomes with sustained release pattern and relatively high SPF.

Keywords: sericin, niosomes, sun protection factor, cream, gel

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Authors: Reshu Saxena, R. K. Tripathi

Abstract:

HIV-1 transmission and spread involves significant host-virus interaction. Potential targets for prevention of HIV-1 lies at the site of mucosal barriers. Thus a better understanding of how HIV-1 infects target cells at such sites and lead their invasion is required, with prime focus on the host determinants regulating HIV-1 spread. HIV-1 Nef is important for viral infectivity and pathogenicity. It promotes HIV-1 replication, facilitating immune evasion by interacting with various host factors and altering cellular pathways via multiple protein-protein interactions. In this study nef was sequenced from HIV-1 patients, and showed specific mutations revealing sequence variability in nef. To explore the difference in Nef functionality based on sequence variability we have studied the effects of HIV-1 Nef in human SupT1 T cell line and (THP-1) monocyte-macrophage cell lines through proteomics approach. 2D-Gel Electrophoresis in control and Nef-transfected SupT1 cells demonstrated several differentially expressed proteins with significant modulation of alpha-enolase. Through further studies, effects of Nef on alpha-enolase regulation were found to be cell lineage-specific, being stimulatory in macrophages/monocytes, inhibitory in T cells and without effect in HEK-293 cells. Cell migration and invasion studies were employed to determine biological function affected by Nef mediated regulation of alpha-enolase. Cell invasion was enhanced in THP-1 cells but was inhibited in SupT1 cells by wildtype nef. In addition, the modulation of enolase and cell invasion remained unaffected by a unique nef variant. These results indicated that regulation of alpha-enolase expression and invasive property of host cells by Nef is sequence specific, suggesting involvement of a particular motif of Nef. To precisely determine this site, we designed a heptapeptide including the suggested alpha-enolase regulating sequence of nef and a nef mutant with deletion of this site. Macrophages/monocytes being the major cells affected by HIV-1 at mucosal barriers, were particularly investigated by the nef mutant and peptide. Both the nef mutant and heptapeptide led to inhibition of enhanced enolase expression and increased invasiveness in THP-1 cells. Together, these findings suggest a possible mechanism of host invasion by HIV-1 through Nef mediated regulation of alpha-enolase and identifies a potential therapeutic target for HIV-1 entry at mucosal barriers.

Keywords: HIV-1 Nef, nef variants, host-virus interaction, tissue invasion

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Authors: Naima Nur, Safa Islam, Saeema Islam, Faridul Alam

Abstract:

Background: Drug-resistant pulmonary tuberculosis (DR-PTB), particularly multidrug-resistant tuberculosis (MDR-TB) and pre-extensive drug-resistant (pre-XDR), is a major challenge in effectively controlling TB, especially in developing. This study aimed to identify the strains of M. tuberculosis complex (MTC) and drug resistance patterns among the pulmonary tuberculosis patients. Methods: The study used a cross-sectional design, and 815 patients were recruited randomly in three study periods. In the first-period, 210 treated PTB patients, who were completed their treatment, received their diagnoses using light microscopy, fluorescence microscopy and cultured on Lowenstein-Jensen (L-J) slant, and then strains were identified as MTC by biochemical tests, and then sensitivity test in National Institute of Diseases of the Chest and Hospital. In the second-period, 220 re-treated PTB patients, who were completed their treatment, received their diagnoses using culture on L-J slant, line probe assay (LPA), and GeneXpert in the same hospital. In the last-period, during treatment, 385 MDR-PTB patients received their diagnoses using culture on L-J slant and LPA in the same hospital. Results: Among sixty-two (29.5%) PTB patients, 13% were sensitive to all first-line anti-TB drugs, 26% were MDR-TB patients, and 14.2% were pre-XDR-TB among 14 MDR-TB patients. After three years, 31% were MDR-TB among 220 re-treated PTB patients. After five years, 16.4% was pre-XDR-TB among 385 MDR-TB patients. Compared to females, male patients were significantly higher at all times. Conclusion: The current study demonstrated that in three study periods, the proportions of DR-TB, MDR-TB, and pre-XDR patients were an alarming issue and increasing daily.

Keywords: multi-drug resistant, drug-resistant, pre-extensive drug resistant, pulmonary tuberculosis

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Authors: Azeez Yusuf, Alan Casey

Abstract:

Silver nanoparticles (AgNP) are one of the most widely investigated metallic nanoparticles due to their promising antibacterial activities. In recent years, AgNP research has shifted beyond antimicrobial use to potential applications in the medical arena. This shift coupled with the extensive commercial applications of AgNP will further increase human exposure, and the subsequent risk of adverse effects that may result from repeated exposures and inefficient delivery meaning research into improved AgNP delivery is of paramount importance. In this study, AgNP were encapsulated in a natural bio-surfactant, dipalmitoylphosphatyidyl choline (DPPC), in an attempt to enhance the intracellular delivery and simultaneously mediate the associated cytotoxicity of the AgNP. It was noted that as a result of the encapsulation, liposomal-AgNP (Lipo-AgNP) at 0.625 μg/ml induced significant cell death in THP1 cell lines a notably lower dose than that of the uncoated AgNP induced cytotoxicity. The induced cytotoxicity was shown to result in an increased level of DNA fragmentation resulting in a cell cycle interruption at the S phase of the cell cycle. It was shown that the predominate form of cell death upon exposure to both uncoated and Lipo-AgNP was apoptosis, however, a ROS-independent activation of the executioner caspases 3/7 occurred when exposed to the Lipo-AgNP. These findings showed that encapsulation of AgNP enhances AgNP cytotoxicity and mediates an ROS-independent induction of apoptosis.

Keywords: silver nanoparticles, AgNP, cytotoxicity, encapsulation, liposome

Procedia PDF Downloads 156

Authors: Nithin Krisshna Gunasekaran, Prathima Prabhu Tumkur, Nicole Nazario Bayon, Krishnan Prabhakaran, Joseph C. Hall, Govindarajan T. Ramesh

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Cerium oxide and turmeric have antioxidant properties, which have gained interest among researchers to study their applications in the field of biomedicine, such asanti-inflammatory, anticancer, and antimicrobial applications. In this study, the turmeric extract was prepared and mixed with cerium nitrate hexahydrate, stirred continuously to obtain a homogeneous solution and then heated on a hot plate to get the supernatant evaporated, then calcinated at 600°C to obtain the cerium oxide nanoparticles. Characterization of synthesized cerium oxide nanoparticles through Scanning Electron Microscopy determined the particle size to be in the range of 70 nm to 250 nm. Energy Dispersive X-Ray Spectroscopy determined the elemental composition of cerium and oxygen. Individual particles were identified through the characterization of cerium oxide nanoparticles using Field Emission Scanning Electron Microscopy, in which the particles were determined to be spherical and in the size of around 70 nm. The presence of cerium oxide was assured by analyzing the spectrum obtained through the characterization of cerium oxide nanoparticles by Fourier Transform Infrared Spectroscopy. The crystal structure of cerium oxide nanoparticles was determined to be face-centered cubic by analyzing the peaks obtained through theX-Ray Diffraction method. The crystal size of cerium oxide nanoparticles was determined to be around 13 nm by using the Debye Scherer equation. This study confirmed the synthesis of cerium oxide nanoparticles using turmeric extract.

Keywords: antioxidant, characterization, cerium oxide, synthesis, turmeric

Procedia PDF Downloads 164

Authors: Hossam E. Emam

Abstract:

The functional textiles development history in the military field could be ascribed as a uniquely interesting research topic. Soldiers are like a high-performance athletes, where monitoring their physical and physiological capabilities is a vital requirement. Functional clothes represent a “second skin” that has a close, “intimate” relationship with the human body. For the application of textiles in military purposes, which is normally required in difficult weather and environmental conditions, several functions are required. The requirements for designing functional military textiles for soldier's protection can be categorized into three categories; i) battle field (protection from chemical warfare agents, flames, and thermal radiation), ii) environmental (water proof, air permeable, UV-protection, antibacterial), iii) physiological (minimize heat stress, low weight, insulative, durability). All of these requirements are important, but the means to fulfill these requirements are not simple and straight forward. Additionally, the combination of more than one function is reported to be very expensive and requires many complicated steps, and the final product is found to be low durability. Not only do all of these requirements are overlapping, but they are also contradicting each other at various levels. Thus, we plan to produce multi-functional textiles (e.g., anti-microbial, UV-protection, fire retardant, photoluminescent) to be applied in military clothes. The current project aims to use quite a simple and applicable technique through the modification of textiles with different coordination polymers and functionalized coordination polymers.

Keywords: functional textiles, military clothes, coordination polymers, antimicrobial, fire retardant, photolumenscent

Procedia PDF Downloads 180

Authors: William E. Bauta, Jennifer Rebeles, Reggie Jacob

Abstract:

Porphyrins are noteworthy in biomedical science for their cancer tissue accumulation and photophysical properties. The preferential accumulation of some porphyrins in cancerous tissue has been known for many years. This, combined with their characteristic photophysical and photochemical properties, including their strong fluorescence and their ability to generate reactive oxygen species in vivo upon laser irradiation, has led to much research into the application of porphyrins as cancer diagnostic and therapeutic agents. Porphyrins have been used as dyes to detect cancer cells both in vivo and, less commonly, in vitro. In one example, human sputum samples from lung cancer patients and patients without the disease were dissociated and stained with the porphyrin TCPP (5,10,15,20-tetrakis-(4-carboxyphenyl)-porphine). Cells were analyzed by flow cytometry. Cancer samples were identified by their higher TCPP fluorescence intensity relative to the no-cancer controls. However, quantitative analysis of fluorescence in cell suspensions stained with multiple fluorophores requires particles stained with each of the individual fluorophores as controls. Fluorescent control particles must be compatible in size with flow cytometer fluidics and have favorable hydrodynamic properties in suspension. They must also display fluorescence comparable to the cells of interest and be stable upon storage amine-functionalized spherical polystyrene beads in the 5 to 20-micron diameter range that was reacted with TCPP and EDC in aqueous pH six buffer overnight to form amide bonds. Beads were isolated by centrifugation and tested by flow cytometry. The 10-micron amine-functionalized beads displayed the best combination of fluorescence intensity and hydrodynamic properties, such as lack of clumping and remaining in suspension during the experiment. These beads were further optimized by varying the stoichiometry of EDC and TCPP relative to the amine. The reaction was accompanied by the formation of a TCPP-related particulate, which was removed, after bead centrifugation, using a microfiltration process. The resultant TCPP-functionalized beads were compatible with flow cytometry conditions and displayed a fluorescence comparable to that of stained cells, which allowed their use as fluorescence standards. The beads were stable in refrigerated storage in the dark for more than eight months. This work demonstrates the first preparation of porphyrin-functionalized flow cytometry control beads.

Keywords: tetraaryl porphyrin, polystyrene beads, flow cytometry, peptide coupling

Procedia PDF Downloads 92

Authors: Tesfay Gebremicheal Reda, K. Samatha, Paul Douglas Sanasi, D. Parajuli

Abstract:

Nanoparticle technology is fast progressing and is being employed in innumerable medical applications. At this time, the public's health is seriously threatened by the rise of bacterial strains resistant to several medications. Metal nanoparticles are a potential alternate approach for tackling this global concern, and this is the main focus of this study. The citrate precursor sol-gel synthesis method was used to synthesize the Niₓ Co₁₋ₓ Fe₂ O₄, (where x = 0.0:0.2:1.0) nanoparticle. XRD identified the development of the cubic crystal structure to have a preferential orientation along (311), and the average particle size was found to be 29-38 nm. The average crystallizes assessed with ImageJ software and origin 22 of the SEM are nearly identical to the XRD results. In the created NCF NPs, the FT-IR spectroscopy reveals structural examinations and the redistribution of cations between octahedral (505-428 cm⁻¹) and tetrahedral (653-603 cm⁻¹) locales. As the Co²⁺ cation is substituted with Ni²⁺, the coercive fields HC decrease from 2384 Oe to 241.93 Oe. Band gap energy rises as Ni concentration increases, which may be attributed to the fact that the ionic radii of Ni²⁺ ions are smaller than that of Co²⁺ ions, which results in a strong electrostatic interaction. On the contrary, except at x = 0.4, the dielectric constant decreases as the nickel concentration increases. According to the findings of this research work, nanoparticles are composed of Ni₀.₄ Co₀.₆ Fe₂ O₄ have demonstrated a promising value against S. aureus and E. coli, and it suggests a proposed model for their potential use as a source of antibacterial agent.

Keywords: antimicrobial, band gap, citrate precursor, dielectric, nanoparticle

Procedia PDF Downloads 73

Authors: R. M. Noah, N. M. Nasir, M. R. Jais, M. S. S. Wahab, M. H. Abdullah, A. S. S. Raj

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Infectious diseases are getting more difficult to treat due to the resistant strains of bacteria. Current generations of antibiotics are most likely ineffective against multi-drug resistant strains bacteria. Thus, there is an urgent need in search of natural antibiotics in particular from medicinal plants. One of the common medicinal plants, Melaleuca cajuputi, has been reported to possess antibacterial properties. The study was conducted to evaluate and justify the presence of antibacterial activity of Melaleuca cajuputi essential oil (EO) against the multi-drug resistant bacteria. Clinical isolates obtained from the teaching hospital were re-assessed to confirm the exact identity of the bacteria to be tested, namely methicillin-resistant staphylococcus aureus (MRSA), carbapenem-resistant enterobacteriaceae (CRE), and extended-spectrum beta-lactamases producer (ESBLs). A well diffusion method was done to observe the inhibition zones of the essential oil against the bacteria. Minimum inhibitory concentration (MIC) was determined using the microdilution method in 96-well flat microplate. The absorbance was measured using a microplate reader. Minimum bactericidal concentration (MBC) was performed using the agar medium method. The zones of inhibition produced by the EO against MRSA, CRE, and ESBL were comparable to that of generic antibiotics used, gentamicin and augmentin. The MIC and MBC results highlighted the antimicrobial efficacy of the EO. The outcome of this study indicated that the EO of Melaleuca cajuputi had antibacterial activity on the multi-drug resistant bacteria. This finding was eventually substantiated by electron microscopy work.

Keywords: melaleuca cajuputi, antibacterial, resistant bacteria, essential oil

Procedia PDF Downloads 122

Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

Abstract:

G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

Procedia PDF Downloads 101

Authors: Jitrawadee Meerasri, Rungsinee Sothornvit

Abstract:

Pectin is a structural polysaccharide from plant cell walls and can be used as a stabilizer, gelling and film-forming agents to improve many food products. Moreover, pectin film as a natural biopolymer can be a carrier of several active ingredients such as antioxidant and antimicrobial to provide an active or functional film. Gamma-aminobutyric acid (GABA) is a well-known agent to reduce neuronal excitability throughout the nervous system and it is interesting to investigate the GABA effect as a substitute of normal plasticizer (glycerol) on edible film properties. Therefore, the objective of this study was to determine the effect of GABA concentrations (5-15% of pectin) on film mechanical properties, moisture content, water vapor permeability, and solubility compared with those from glycerol (10% of pectin) plasticized pectin film including a control film (pectin film without any plasticizer). It was found that an increase in GABA concentrations decreased film tensile strength, modulus, solubility and water vapor permeability, but elongation was increased without a change in the moisture content. The smaller amount of GABA showed the equivalent film properties as using a higher amount of glycerol. Consequently, GABA can act as an alternative plasticizer substitute of glycerol at the lower amount used. Moreover, GABA provides the nutritional high value in the food products when the edible packaging material is consumed with products.

Keywords: gamma-aminobutyric acid, pectin, plasticizer, edible film

Procedia PDF Downloads 130

Authors: Kiran Shehzadi, Yasmeen Akhtar, Mujahid Ameen, Tabinda Ijaz, Shoukat Siddique

Abstract:

Nanoparticles, due to their different sizes and morphologies, are employed in various fields such as the medical field, cosmetics, pharmaceutical, textile industry as well as in paints, adhesives, and electronics. Metal nanoparticles exhibit excellent antimicrobial activity, dye degradation and can be used as anti-cancerous drug loading agents. In this study, sZilver nanoparticles (Ag-NPs) were synthesized employing doxycycline (antibiotic) as a reducing and capping agent (biological/green synthesis). Produced Ag-NPS were characterized using UV/VIS spectrophotometry, XRD, SEM, and FTIR. Surface plasmon resonance (SPR) of silver nanoparticles was observed at 411nm with 90nm size with homogenized spherical shape. These particles revealed good inhibition zones for Fungi such as Candida albicans and Candida tropicalis. In this study, toxic properties of Ag-NPs were monitored by allowing them to penetrate in the cell, causing an abrupt increase in oxidative stress, which resulted ultimately in cell death. Histopathological analysis of mice organs was performed by administering definite concentrations of silver nanoparticles orally to mice for 14 days. Toxic properties were determined, and it was revealed that the toxicity of silver nanoparticles mainly depends on the size. Silver nanoparticles of this work presented mild toxicity for different organs (liver, kidney, spleen, heart, and stomach) of mice.

Keywords: metal nanoparticles, green/biological methods, toxicity, Candida albicans, Candida tropicalis

Procedia PDF Downloads 129

Authors: Yilei Xue, Yat-Hing Ham, Wenting Qiu, Wan Chan, Stefan Nagl

Abstract:

Polydopamine (PDA) is a popular material in biological and medical applications due to its excellent biocompatibility, outstanding physicochemical properties, and facile fabrication. In this project, a new sandwich-structured PDA and silver (Ag) hybrid material named PDA-Ag-PDA was synthesized and characterized layer-by-layer, where silver nanoparticles (Ag NPs) are wrapped in PDA coatings, using SEM, AFM, 3D surface metrology, and contact angle meter. The silver loading capacity is positively proportional to the roughness value of the initial PDA film. This designed film was subsequently integrated within a digital microfluidic (DMF) platform coupling with an oxygen sensor layer for on-chip antibacterial assay. The concentration of E. coli was quantified on DMF by real-time monitoring oxygen consumption during E. coli growth with the optical oxygen sensor layer. The PDA-Ag-PDA coating shows an 99.9% reduction in E. coli population under non-nutritive condition with 1-hour treatment and has a strong growth inhibition of E. coliin nutrient LB broth as well. Furthermore, PDA-Ag-PDA film maintaining a low cytotoxicity effect to human cells. After treating with PDA-Ag-PDA film for 24 hours, 82% HEK 293 and 86% HeLa cells were viable. The SERS enhancement factor of PDA-Ag-PDA is estimated to be 1.9 × 104 using Rhodamine 6G (R6G). Multifunctional PDA-Ag-PDA coating provides an alternative platform to conjugate biomolecules and perform biological applications on DMF, in particular, for the adhesive protein and cell study.

Keywords: polydopamine, silver nanoparticles, digital microfluidic, optical sensor, antimicrobial assay, SERS

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Authors: Sarekha Woranuch, Rangrong Yoksan

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Active packaging has been developed based on the incorporation of certain additives, in particular antimicrobial and antioxidant agents, into packaging systems to maintain or extend product quality and shelf-life. Ferulic acid is one of the most effective natural phenolic antioxidants, which has been used in food, pharmaceutical and active packaging film applications. However, most phenolic compounds are sensitive to oxygen, light and heat; its activities are thus lost during product formulation and processing. Grafting ferulic acid onto polymer is an alternative to reduce its loss under thermal processes. Therefore, the objectives of the present research were to study the thermal stability of ferulic acid after grafting onto chitosan, and to investigate the possibility of using ferulic acid-grafted chitosan (FA-g-CTS) as an antioxidant for active biodegradable packaging film. FA-g-CTS was incorporated into biodegradable film via a two-step process, i.e. compounding extrusion at temperature up to 150 °C followed by blown film extrusion at temperature up to 175 °C. Although incorporating FA-g-CTS with a content of 0.02–0.16% (w/w) caused decreased water vapor barrier property and reduced extensibility, the films showed improved oxygen barrier property and antioxidant activity. Radical scavenging activity and reducing power of the film containing FA-g-CTS with a content of 0.04% (w/w) were higher than that of the naked film about 254% and 94%, respectively. Tensile strength and rigidity of the films were not significantly affected by adding FA-g-CTS with a content of 0.02–0.08% (w/w). The results indicated that FA-g-CTS could be potentially used as an antioxidant for active packaging film.

Keywords: active packaging film, antioxidant activity, chitosan, ferulic acid

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Authors: Iman A. A. Kassem, Ayman A. Farghaly, Zeinab M. Hassan, Farouk R. Melek, Neveen S. Ghaly

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Lipidium sativum L, an annual herb that grows to 50 cm, is known as an important member of family Brassicaceae. Besides its nutritional value, the seeds were widely used in folk medicine for treatment of cough, asthma, and headache. It was also reported to possess hypocholesterolemic, anti-inflammatory, antidiarrheal, antimicrobial and anticancer activities. In this study, the genoprotective properties of L. sativum seed methanolic extract (LSME) were evaluated in vivo. Three groups of mice were given LSME for five consecutive days at the three dose levels 25, 50 and 100 mg/kg b.wt. The three groups were then injected intraperitoneally with cyclophosphamide at a dose of 20 mg/kg b.wt. to induce DNA damage. A group received only cyclophosphamide (20 mg/kg b.wt.) served as control. LSME significantly inhibited the DNA aberrations in mice caused by cyclophosphamide in a dose-dependent manner in the two groups that received LSME at 50 and 100 mg/kg b.wt. dose levels. The chromosomal aberrations' inhibitory indices were calculated as 18 and 31 in mice bone marrow cells and 27 and 48 in mice spermatocytes, respectively. Phytochemical examination carried out by us revealed that flavonoids were the main chemical constituents of LSME. The major flavonoids kaempferol, kaempferol-3-O-rhamnoside, kaempferol-3-O-glucoside, quercetin, and quercetin-3-O-glucoside were isolated and characterized. It was concluded that the genoprotective effect of LSME might be attributed to the presence of flavonoids which are well-known for their antioxidant properties.

Keywords: cyclophosphamide, flavonoids, genoprotective effect, Lepidium sativum

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Authors: Anika Pallapothu

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Periodontitis, a severe gum disease, poses a significant threat to the integrity of bone and soft tissues supporting teeth, primarily initiated by bacterial accumulation around the gum line. Conventional treatments like scaling/root planning and plaque removal are widely employed, but integrating modern technologies such as nanotechnology holds promise for innovative therapeutic approaches. This study explores the utilization of silver nanoparticles encapsulated within cellulose nanofiber (CNF) and mesoporous bioactive glass hydrogel matrices for periodontitis management. Silver nanoparticles exhibit potent antimicrobial properties by disrupting microbial cell membranes, inducing reactive oxygen species (ROS) generation, and interfering with vital cellular processes like ATP production and nucleic acid synthesis. Mesoporous bioactive glass, renowned for its high surface area, osteoconductive, and bioactivity, presents a favorable platform for pharmaceutical applications. Incorporating CNF enhances the properties of the hydrogel due to its biocompatibility, biodegradability, and water absorption capacity. The proposed composite material is anticipated to exert beneficial effects in periodontitis treatment by demonstrating antibacterial and anti-inflammatory activities, offering a promising avenue for future therapeutic interventions.

Keywords: periodontitis, cellulose nanofibers, silver nanoparticles, mesoporous bioactive glass, antibacterial activity, anti-inflammatory activity

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Authors: Rajesh Kumar Meena, Suman Jhajharia, Goutam Chakraborty

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Plasmonic silver nanoparticles (Ag NPs) was synthesized by chemical reduction method using Trachyspermum Ammi (TA, Ajwain) seeds extract in aqueous medium and AgNO3 solution at different time interval. Reaction time, and concentration of AgNO3 and TA could accelerate the reduction rate of Ag+ and affect AgNPs size and concentration of NPs. Surface plasmon resonance band centered at 420-430 nm (88.78nm) was recognised as first exitonic peak of UV-Vis absorption spectra of AgNPs that used to calculate the particle size (10-30 nm). FTIR results TA supported AgNPs showed decrease in intensity of peaks at 3394, 1716 and 1618 cm-1 with respect to the plain TA indicating the involvement of O-H, carbonyl group and C=C stretching in formation of TA-AgNPs aggregates. The C-O-C and C-N stretching suggested the presence of many phytochemicals on the surface of the NPs. Impedance study reveals that at low concentration of TA the rate of charge transfer is in TA-AgNPs aggregates, found higher than the higher TA concentration condition that confirms the stability of AgNPs in water. Extract reduce silver ions into silver nanoparticles (NPs) of size 6-50nm. Pronounce effect of the time on Ag NPs concentration and particle size, was exhibited by the system These biogenic Ag NPs are characterized using UV- Vis spectrophotometry (UV-Visible), Fourier transformation infrared (FTIR) and XRD. These studies give us inside view of the most probable mechanism of biosynthesis and optoelectronic properties of the as synthesised Ag NPs.

Keywords: antimicrobial activity, bioreduction, capping agent, silver nanoparticles

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Authors: Sandip Shrestha, Ann M. Donoghue, Komala Arsi, Basanta R. Wagle, Abhinav Upadhyay, Dan J. Donoghue

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Campylobacter, one of the major cause of foodborne illness worldwide, is commonly present in the intestinal tract of poultry. Many strategies are currently being investigated to reduce Campylobacter counts on commercial poultry during processing with limited success. This study investigated the efficacy of the generally recognized as safe compound, carvacrol (CR), a component of wild oregano oil as a wash treatment for reducing C. jejuni and aerobic bacteria on chicken skin. A total of two trials were conducted, and in each trial, a total of 75 skin samples (4cm × 4cm each) were randomly allocated into 5 treatment groups (0%, 0.25%, 0.5%, 1% and 2% CR). Skin samples were inoculated with a cocktail of four wild strains of C. jejuni (~ 8 log10 CFU/skin). After 30 min of attachment, inoculated skin samples were dipped in the respective treatment solution for 1 min, allowed to drip dry for 2 min and processed at 0, 8, 24 h post treatment for enumeration of C. jejuni and aerobic bacterial counts (n=5/treatment/time point). The data were analyzed by ANOVA using PROC GLM procedure of SAS 9.3. All the tested doses of CR suspension consistently reduced C. jejuni counts across all time points. The 2% CR wash was the most effective treatment and reduced C. jejuni counts by ~4 log₁₀ CFU/sample (P < 0.05). Aerobic counts were reduced for the 0.5% CR dose at 0 and 24h in Trial 1 and at 0, 8 and 24h in Trial 2. The 1 and 2% CR doses consistently reduced aerobic counts in both trials up to 2 log₁₀ CFU/skin.

Keywords: Campylobacter jejuni, carvcrol, chicken skin, postharvest

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Authors: Abed Hannane, Rouag Noureddine

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The present study aims to evaluate the association of fresh garlic (Allium sativum L.) and storage at 4°C in preserving the microbiological, nutritional, and sanitary quality of Merguez-type sausages prepared and sold locally from meat offal. The analysis focused on the evaluation of the microbiological quality of fifteen samples randomly taken from several butcheries in the wilaya of BBA, eastern Algeria. The bacteriological analysis revealed the presence of 6.88.10⁵ CFU/g of total aerobic bacteria, 5.39.10⁵ CFU/g of total coliforms, 2.23.10⁵ CFU/g of fecal coliforms, 2.43.103 CFU/g of Escherichia coli and 1.8.10⁵ CFU/g of coagulase-positive staphylococci, values higher than Algerian standards. The addition of fresh garlic as an antibacterial preservative at concentrations of 0.06, 0.12, 0.18, and 0.24 g/g to ground beef samples and stored in the refrigerator at 4°C for 15 days. The addition of garlic to Merguez made it possible to significantly reduce the presence of different bacterial groups during their refrigerated storage, compared to untreated meat, bringing it below the standards defined in the matter. Thus, the use of garlic as a food additive at a concentration of 0.12 g/g was sufficient to obtain levels according to Algerian standards equal to 1.8.10⁴ CFU/g of total aerobic bacteria, 9.48.10³ CFU/ g of total coliforms, 3.68.10³ UFC/g fecal coliforms, 4.56.10² UFC/g of E.coli 2.39.10⁴ UFC/g of coagulase-positive staphylococci. It is clear that thanks to the addition of garlic to Merguez, the sanitary quality has been improved by reducing the aerobic bacterial load and increasing the shelf life at 4°C.

Keywords: antimicrobial effect, garlic, sausage, storage

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Authors: Zainab M. Almutairi, Amjad Alharbi

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The use of engineered nanomaterials has increased as a result of their positive impact on many sectors of the economy, including agriculture. Silver nanoparticles (AgNPs) are now used to enhance seed germination, plant growth, and photosynthetic quantum efficiency and as antimicrobial agents to control plant diseases. In this study, we examined the effect of AgNP dosage on the seed germination of three plant species: corn (Zea mays L.), watermelon (Citrullus lanatus [Thunb.] Matsum. & Nakai) and zucchini (Cucurbita pepo L.). This experiment was designed to study the effect of AgNPs on germination percentage, germination rate, mean germination time, root length and fresh and dry weight of seedlings for the three species. Seven concentrations (0.05, 0.1, 0.5, 1, 1.5, 2, and 2.5 mg/ml) of AgNPs were examined at the seed germination stage. The three species had different dose responses to AgNPs in terms of germination parameters and the measured growth characteristics. The germination rates of the three plants were enhanced in response to AgNPs. Significant enhancement of the germination percentage values was observed after treatment of the watermelon and zucchini plants with AgNPs in comparison with untreated seeds. AgNPs showed a toxic effect on corn root elongation, whereas watermelon and zucchini seedling growth were positively affected by certain concentrations of AgNPs. This study showed that exposure to AgNPs caused both positive and negative effects on plant growth and germination.

Keywords: citrullus lanatus, cucurbita pepo, seed germination, seedling growth, silver nanoparticles, zea mays

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Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

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Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

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Authors: Indrajeet Kumar, Jayanta Bhattacharya

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This study was evaluated for the potential use of nanosilver (nAg) as a disinfectant and antimicrobial growth promoter supplement for the poultry. The experiments were conducted in the Kangsabati river basin region, in West Medinipur district, West Bengal, India for six months. Two poultry farms were adopted for the experiment. The rural economy of this region from Jhargram to Barkola is heavily dependent on contract poultry farming. The water samples were collected from the water source of poultry farm which has been used for poultry drinking purpose. The bacteriological analysis of water sample revealed that the total bacterial count (total coliform and E. coli) were higher than the acceptable standards. The bacterial loads badly affected the growth performance and health of the poultry. For disinfection, a number of chemical compounds (like formaldehyde, calcium hypochloride, sodium hypochloride, and sodium bicarbonate) have been used in typical commercial formulations. However, the effects of all these chemical compounds have not been significant over time. As a part of our research-to-market initiative, we used nanosilver (nAg) formulation as a disinfectant. The nAg formulation was synthesized by hydrothermal technique and characterized by UV-visible, TEM, SEM, and EDX. The obtained results revealed that the mortality rate of poultry was reduced due to nAg formulation compared to the mortality rate of the negative control. Moreover, the income of the farmer family was increased by 10-20% due to less mortality and better health of the poultry.

Keywords: farm water, nanosilver, field application, and poultry performance

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Authors: Mary Anthony Oguike, Ilouno, Amaduruonye

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A completely randomized design experiment with 3 treatments was conducted to study the effects of garlic on intestine characteristics, haematology and serum biochemistry of Marshal broilers. Thirty three (33) broiler chicks were randomly allotted to each treatment designated T1, T2 and T3. The birds in each treatment were replicated 3 times with 11 broilers per replicate. They were fed diets supplemented with garlic at 0, 1.5 and 2.5 % /kg feed for t1, T2 and T3, respectively with T1 as control. Data were collected on intestine parameters, serum biochemical parameters and haematological indices. The results showed significant (P>0.05) dose-dependent decrease in intestine weight and caeca microbial load of the broilers. The intestine of broilers in the treatments showed normal histological architecture in all the treatments. The red blood cell (RBC), white blood cell (WBC), haemoglobin (Hb) and other haematological indices showed no significant differences (P<0.05) among the treatments. Cholesterol, globulin, glucose and alanin aminotransferase (ALT) were significantly different (P<0.05) among the treatment groups. Serum biochemical parameters such as, total protein albumin, bilirubin and others were not significant among the treatments. All the blood parameters studied fall within the normal range for broilers. Garlic supplementation in the diets of broilers did not have any detrimental effects on the treated birds since their serum biochemistry and haematology fall within the normal range for broilers birds. The microbial examination of intestine and caeca, as well as the histopathological studies of the intestine confirmed antimicrobial properties of garlic.

Keywords: broiler, biochemistry and haematology, garlic, intestine

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