Search results for: phenol biosensor

Authors: Darshna Sharma, Suban K. Sahoo

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The detection of intracellular fluoride in human cancer cell HeLa was achieved by chemosensors derived from vitamin B6 cofactors using fluorescence imaging technique. These sensors were first synthesized by condensation of pyridoxal/pyridoxal phosphate with 2-amino(thio)phenol. The anion recognition ability was explored by experimental (UV-VIS, fluorescence and 1H NMR) and theoretical DFT [(B3LYP/6-31G(d,p)] methods in DMSO and mixed DMSO-H2O system. All the developed sensors showed both naked-eye detectable color change and remarkable fluorescence enhancement in the presence of F- and AcO-. The anion recognition was occurred through the formation of hydrogen bonded complexes between these anions and sensor, followed by the partial deprotonation of sensor. The detection limit of these sensors were down to micro(nano) molar level of F- and AcO-.

Keywords: chemosensors, fluoride, acetate, turn-on, live cells imaging, DFT

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Authors: Toru Shionoya, Yosuke Kurihara, Takashi Kaburagi, Kajiro Watanabe

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Mass-produced bidirectional microphones have attractive characteristics. They work as a microphone as well as a sensor with high gain over a wide frequency range; they are also highly reliable and economical. We present novel multiple functional uses of the microphones. A mathematical model for explaining the high-pass-filtering characteristics of bidirectional microphones was presented. Based on the model, the characteristics of the microphone were investigated, and a novel use for the microphone as a sensor with a wide frequency range was presented. In this study, applications for using the microphone as a security sensor and a human biosensor were introduced. The mathematical model was validated through experiments, and the feasibility of the abovementioned applications for security monitoring and the biosignal monitoring were examined through experiments.

Keywords: bidirectional microphone, low-frequency, mathematical model, frequency response

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Authors: Thi Huong Vu, Vijay Jayasena, Zhongxiang Fang, Gary Dykes

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D-pinitol (3-O-methyl ether of D-chiro-inosito) has been known to have health benefits for diabetic patients. Carob kibble has received attention due to the presence of high value D-pinitol and polyphenol antioxidants. D-pinitol was concentrated from carob kibble using submerged fermentation with Saccharomyces cerevisiae. Total carbohydrates and D-pinitol were determined by the phenol-sulphuric acid method and HPLC, respectively. The content of D-pinitol increased from approximately 43 to 70 mg/g dry weight after fermentation. The yeast consumed over 70% of total carbohydrates in carob kibble without any negative effect on D-pinitol content. A range of substrate medium pH’s from 5.0 – 7.0 had no significant effect on the removal of carbohydrates and D-pinitol. This method may provide a practical solution for production of D-pinitol from carob in a cost effective manner.

Keywords: carob kibble, d-pinitol, saccharomyces cerevisiae, submerged fermentation, total carbohydrates

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Authors: Ahmad Manbohi, Seyyed Hamid Ahmadi

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A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R^² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.

Keywords: biological fluids, biomarkers, microfluidic paper-based electrochemical biosensors, Multiplex

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Authors: Mustapha Mahmoud Dif, Fouzia Benali-Toumi, Mohamed Benyahia, Sofiane Bouazza, Abbes Dellal, Slimane Baha

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L. multifida is applied to treat rheumatism and cold and has hypoglycemic and anti-inflammatory properties. The present study is to optimize the extraction of phenolic compounds in Algerian Lavandula multifida. The influences of parameters including temperature (decoction and maceration) and extraction time (15min to 45 min) on the flavonoids concentration are studied. The optimal conditions are determined and the quadratic response surfaces draw from the mathematical models. Total phenols were evaluated using Folin sicaltieu methods, total flavonoids were estimated using the Tri chloral aluminum method. The maximum concentration extracted, for total flavonoids, equal to 0.043 mg/g was achieved with decoction and extraction time of 41.55 min. However, for total phenol compounds highest concentration of 0.218 mg/g, is obtained with 45 min at 49.99°C.

Keywords: L multifidi, phenolic content, optimization, time, temperature

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Authors: M. Salvador, J. C. Martinez-Garcia, A. Moyano, M. C. Blanco-Lopez, M. Rivas

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Biosensors play a crucial role in the detection of molecules nowadays due to their advantages of user-friendliness, high selectivity, the analysis in real time and in-situ applications. Among them, Lateral Flow Immunoassays (LFIAs) are presented among technologies for point-of-care bioassays with outstanding characteristics such as affordability, portability and low-cost. They have been widely used for the detection of a vast range of biomarkers, which do not only include proteins but also nucleic acids and even whole cells. Although the LFIA has traditionally been a positive/negative test, tremendous efforts are being done to add to the method the quantifying capability based on the combination of suitable labels and a proper sensor. One of the most successful approaches involves the use of magnetic sensors for detection of magnetic labels. Bringing together the required characteristics mentioned before, our research group has developed a biosensor to detect biomolecules. Superparamagnetic nanoparticles (SPNPs) together with LFIAs play the fundamental roles. SPMNPs are detected by their interaction with a high-frequency current flowing on a printed micro track. By means of the instant and proportional variation of the impedance of this track provoked by the presence of the SPNPs, quantitative and rapid measurement of the number of particles can be obtained. This way of detection requires no external magnetic field application, which reduces the device complexity. On the other hand, the major limitations of LFIAs are that they are only qualitative or semiquantitative when traditional gold or latex nanoparticles are used as color labels. Moreover, the necessity of always-constant ambient conditions to get reproducible results, the exclusive detection of the nanoparticles on the surface of the membrane, and the short durability of the signal are drawbacks that can be advantageously overcome with the design of magnetically labeled LFIAs. The approach followed was to coat the SPIONs with a specific monoclonal antibody which targets the protein under consideration by chemical bonds. Then, a sandwich-type immunoassay was prepared by printing onto the nitrocellulose membrane strip a second antibody against a different epitope of the protein (test line) and an IgG antibody (control line). When the sample flows along the strip, the SPION-labeled proteins are immobilized at the test line, which provides magnetic signal as described before. Preliminary results using this practical combination for the detection and quantification of the Prostatic-Specific Antigen (PSA) shows the validity and consistency of the technique in the clinical range, where a PSA level of 4.0 ng/mL is the established upper normal limit. Moreover, a LOD of 0.25 ng/mL was calculated with a confident level of 3 according to the IUPAC Gold Book definition. Its versatility has also been proved with the detection of other biomolecules such as troponin I (cardiac injury biomarker) or histamine.

Keywords: biosensor, lateral flow immunoassays, point-of-care devices, superparamagnetic nanoparticles

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Authors: Ehsan Yazdanpanah Moghadam, Muthukumaran Packirisamy

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In the present study, a design of the suspended polymeric microfluidic platform is introduced that is fabricated with three polymeric layers. Changing the microchannel plane to be perpendicular to microcantilever plane, drastically decreases moment of inertia in that direction. In addition, the platform is made of polymer (around five orders of magnitude less compared to silicon). It causes significant increase in the sensitivity of the cantilever deflection. Next, although the dimensions of this platform are constant, by misaligning the embedded microchannels laterally in the suspended microfluidic platform, the sensitivity can be highly increased. The investigation is studied on four fluids including water, seawater, milk, and blood for flow ranges from low rate of 5 to 70 µl/min to obtain the best design with the highest sensitivity. The best design in this study shows the sensitivity increases around 50% for water, seawater, milk, and blood at the flow rate of 70 µl/min by just misaligning the embedded microchannels in the suspended polymeric microfluidic platform.

Keywords: microfluidic, MEMS, biosensor, microresonator

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Authors: M. Simchi, M. Amiri, E. Rezvani, I. Mirzaei, M. Berahman, A. Simchi, M. Fardmanesh

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Graphene is a single-atomic two-dimensional crystal of carbon atoms that has considerable properties due to its unique structure and physics with applications in different fields. Graphene has sensitive electrical properties due to its atomic-thin structure. Along with the substrate materials and their influence on the transport properties in graphene, design and fabrication of graphene-based devices for biomedical and biosensor applications are challenging. In this work, large-area high-quality graphene nanosheets were prepared by low pressure chemical vapor deposition using methane gas as carbon source on copper foil and transferred on the biocompatible substrates. Through deposition of titanium and gold contacts, current-voltage response of the transferred graphene on four biocompatible substrates, including PDMS, SU-8, Nitrocellulose, and Kapton (Fig. 2) were experimentally determined. The considerable effect of the substrate type on the electrical properties of graphene is shown. The sheet resistance of graphene is changed from 0.34 to 14.5 kΩ/sq, depending on the substrate.

Keywords: biocompatible substrates, electrical properties, graphene, sheet resistance

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Authors: Ewelina Grabowska, Martyna Marchelek

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Among the various semiconductors, nanosized TiO2 has been widely studied due to its high photosensitivity, low cost, low toxicity, and good chemical and thermal stability. However, there are two main drawbacks to the practical application of pure TiO2 films. One is that TiO2 can be induced only by ultraviolet (UV) light due to its intrinsic wide bandgap (3.2 eV for anatase and 3.0 eV for rutile), which limits its practical efficiency for solar energy utilization since UV light makes up only 4-5% of the solar spectrum. The other is that a high electron-hole recombination rate will reduce the photoelectric conversion efficiency of TiO2. In order to overcome the above drawbacks and modify the electronic structure of TiO2, some semiconductors (eg. CdS, ZnO, PbS, Cu2O, Bi2S3, and CdSe) have been used to prepare coupled TiO2 composites, for improving their charge separation efficiency and extending the photoresponse into the visible region. It has been proved that the fabrication of p-n heterostructures by combining n-type TiO2 with p-type semiconductors is an effective way to improve the photoelectric conversion efficiency of TiO2. SrTiO3 is a good candidate for coupling TiO2 and improving the photocatalytic performance of the photocatalyst because its conduction band edge is more negative than TiO2. Due to the potential differences between the band edges of these two semiconductors, the photogenerated electrons transfer from the conduction band of SrTiO3 to that of TiO2. Conversely, the photogenerated electrons transfer from the conduction band of SrTiO3 to that of TiO2. Then the photogenerated charge carriers can be efficiently separated by these processes, resulting in the enhancement of the photocatalytic property in the photocatalyst. Additionally, one of the methods for improving photocatalyst performance is addition of nanoparticles containing one or two noble metals (Pt, Au, Ag and Pd) deposited on semiconductor surface. The mechanisms were proposed as (1) the surface plasmon resonance of noble metal particles is excited by visible light, facilitating the excitation of the surface electron and interfacial electron transfer (2) some energy levels can be produced in the band gap of TiO2 by the dispersion of noble metal nanoparticles in the TiO2 matrix; (3) noble metal nanoparticles deposited on TiO2 act as electron traps, enhancing the electron–hole separation. In view of this, we recently obtained series of TiO2@SrTiO3 and SrTiO3 photocatalysts loaded with noble metal NPs. using photodeposition method. The M- TiO2@SrTiO3 and M-SrTiO3 photocatalysts (M= Rh, Rt, Pt) were studied for photodegradation of phenol in aqueous phase under UV-Vis and visible irradiation. Moreover, in the second part of our research hydroxyl radical formations were investigated. Fluorescence of irradiated coumarin solution was used as a method of ˙OH radical detection. Coumarin readily reacts with generated hydroxyl radicals forming hydroxycoumarins. Although the major hydroxylation product is 5-hydroxycoumarin, only 7-hydroxyproduct of coumarin hydroxylation emits fluorescent light. Thus, this method was used only for hydroxyl radical detection, but not for determining concentration of hydroxyl radicals.

Keywords: composites TiO2, SrTiO3, photocatalysis, phenol degradation

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Authors: Bo Ram Choi, Ji Su Kim, Juyeon Cho, Hyukjin Lee

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Food contaminated by bacteria (E.coli), causes food poisoning, which occurs to many patients worldwide annually. We have introduced an application of rolling circle amplification (RCA) as a versatile biosensor and developed a diagnostic platform composed of capillary tube and microbeads for rapid and easy detection of Escherichia coli (E. coli). When specific mRNA of E.coli is extracted from cell lysis, rolling circle amplification (RCA) of DNA template can be achieved and can be visualized by beads aggregation in capillary tube. In contrast, if there is no bacterial pathogen in sample, no beads aggregation can be seen. This assay is possible to detect visually target gene without specific equipment. It is likely to the development of a genetic kit for point of care testing (POCT) that can detect target gene using microbeads.

Keywords: rolling circle amplification (RCA), Escherichia coli (E. coli), point of care testing (POCT), beads aggregation, capillary tube

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Authors: Isa Usman Balan, Umar Aliyu, Ahmad Tijjani Muhammed

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The laboratory scale experiment was conducted to determine the phytochemical constituents and antibacterial activities of epiphytic neem leaves (Tapinanthusdodoneifolius) extracts on some selected clinical isolates. The samples were collected using polythene bags to avoid unnecessary contamination of the plants, and they were collected from the old site garden of the BUK. The phytochemical screening and antibacterial test were carried out in the Chemistry and Biology laboratory, respectively at Bayero University Kano (BUK). The result obtained showed that carbohydrates, glycosides, steroids, alkaloids, phenol, saponins and flavonoids are present in the ethanolic extract. However, chloroform extract showed only glycosides, phenols, and carbohydrates. Furthermore, there was no significant difference between the ethanolic extracts and bacterial isolates (p<0.05).

Keywords: phytochemical screening, antibacterial, clinical isolates, epiphytic neem leaves, Tapinanthus dodoneifolius

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Authors: Tanwi Priya, Brijesh Kumar Mishra

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Occurrence of aromatic fractions of Natural Organic Matter (NOM) led to formation of carcinogenic disinfection by products such as trihalomethanes in chlorinated water. In the present study, the efficiency of aluminium based electrocoagulation on the removal of prominent aromatic groups such as phenol, hydrophobic auxochromes, and carboxyl groups from NOM enriched synthetic water has been evaluated using various spectral indices. The effect of electrocoagulation on turbidity has also been discussed. The variation in coagulation performance as a function of pH has been studied. Our result suggests that electrocoagulation can be considered as appropriate remediation approach to reduce trihalomethanes formation in water. It has effectively reduced hydrophobic fractions from NOM enriched low turbid water. The charge neutralization and enmeshment of dispersed colloidal particles inside metallic hydroxides is the possible mechanistic approach in electrocoagulation.

Keywords: aromatic fractions, electrocoagulation, natural organic matter, spectral indices

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Authors: Mustofa Ahda, Fiqri Rozi, Gina Noor Habibah, Mas Ulfah Lestari, Tomy Hardianto, Yuni Andriani

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The development of natural antioxidants in the Centella asiatica and Majapahit is a great potential. This research has been optimizing the composition of ethanol extract of Centella asiatica and leaves Majapahit as an antioxidants source using measure the free radical scavenging activity of DPPH. The results of the research showed that both the ethanol extract of Centella asiatica and leaves Majapahit has a total content of phenol. It is shown with the ability to reduce reagent Folin Ciocalteu become blue colour. The composition optimization of extract Centella asiatica leaves Majapahit = 30:70 has free radical scavenging activity of DPPH most well compared ethanol extract of Centella asiatica and leaves Majapahit. IC50 values for the composition of ethanol extract of Centella asiatica : leaves Majapahit = 30:70 is 0,103 mg/mL.

Keywords: antioxidant activity, Centella asiatica, Cresentia cujete, composition extract

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Authors: Lekha Logu

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The coordination behavior of the newly synthesized Schiff base ligands, 4-bromo-2-((p-tolyl imino) methyl) phenol obtained by condensing para-toluidine with 5-bromo salicylaldehyde and N-(3,4-dichloro benzylidene)-4-methylbenzenamine obtained by condensing Para-toluidine with 3,4-dichloro benzaldehyde in ethanolic medium has been explored in this current study. The synthesized Schiff’s base ligands were complexed with lanthanide nitrate salts yielding [LnL(NO3)2(H2O)2]NO3, (Ln=Pr, Sm). Elemental analysis, conductance measurement, and spectral techniques like Nuclear Magnetic Resonance (NMR), Ultraviolet-visible (UV-Vis) and Fourier Transform Infrared (FTIR) have been used to characterize Schiff’s base ligands and their lanthanide metal complexes. An attempt has been made on these complexes for their antimicrobial activity against the gram-positive and gram-negative bacterial species like Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Klebsiella pneumonia and fungal species like Canadida and Aspergillus.

Keywords: lanthanide complexes, Schiff's base, antimicrobial assay, synthesis, characterization

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Authors: Muhammad Rosli Abdullah, Noor Hasmiza Harun

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A microorganism detection system has a potential to be used with the advancement in a biosensor development. The detection system requires an optical sensing system, microfluidic device and biological reagent. Although, the biosensors are available in the market, a label free and a lab-on-chip approach will promote a flexible solution. As a preliminary study of microorganism detection, three mechanisms such as Total Internal Reflection (TIR), Micro Fluidic Channel (MFC) and magnetic-electric field propagation were study and simulated. The objective are to identify the TIR angle, MFC parabolic flow and the wavelength for the microorganism detection. The simulation result indicates that evanescent wave is achieved when TIR angle > 42°, the corner and centre of a parabolic velocity are 0.02 m/s and 0.06 m/s respectively, and a higher energy distribution of a perfect electromagnetic scattering with dipole resonance radiation occurs at 500 nm. This simulation is beneficial to determine the components of the microorganism detection system that does not rely on classical microbiological, immunological and genetic methods which are laborious, time-consuming procedures and confined to specialized laboratories with expensive instrumentation equipment.

Keywords: microorganism, microfluidic, total internal reflection, lab on chip

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Authors: Cheng-Cao Sun, Shu-Jun Li, De-Jia Li

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Determinants of growth and metastasis in cancer remain of great interest to define. MicroRNAs (miRNAs) have frequently emerged as tumor metastatic regulator by acting on multiple signaling pathways. Here, we report the definition of miR-346 as an oncogenic microRNA that facilitates non-small cell lung cancer (NSCLC) cell growth and metastasis. XPC, an important DNA damage recognition factor in nucleotide excision repair was defined as a target for down-regulation by miR-346, functioning through direct interaction with the 3'-UTR of XPC mRNA. Blocking miR-346 by an antagomiR was sufficient to inhibit NSCLC cell growth and metastasis, an effect that could be phenol-copied by RNAi-mediated silencing of XPC. In vivo studies established that miR-346 overexpression was sufficient to promote tumor growth by A549 cells in xenografts mice, relative to control cells. Overall, our results defined miR-346 as an oncogenic miRNA in NSCLC, the levels of which contributed to tumor growth and invasive aggressiveness.

Keywords: microRNA-346, miR-346, XPC, non-small cell lung cancer, oncogenesis

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Authors: Marta Paszkiewicz, Justyna Łuczak, Martyna Marchelek, Adriana Zaleska-Medynska

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In recent years, photocatalytical processes have been intensively investigated for destruction of pollutants, hydrogen evolution, disinfection of water, air and surfaces, for the construction of self-cleaning materials (tiles, glass, fibres, etc.). Titanium dioxide (TiO2) is the most popular material used in heterogeneous photocatalysis due to its excellent properties, such as high stability, chemical inertness, non-toxicity and low cost. It is well known that morphology and microstructure of TiO2 significantly influence the photocatalytic activity. This characteristics as well as other physical and structural properties of photocatalysts, i.e., specific surface area or density of crystalline defects, could be controlled by preparation route. In this regard, TiO2 particles can be obtained by sol-gel, hydrothermal, sonochemical methods, chemical vapour deposition and alternatively, by ionothermal synthesis using ionic liquids (ILs). In the TiO2 particles synthesis ILs may play a role of a solvent, soft template, reagent, agent promoting reduction of the precursor or particles stabilizer during synthesis of inorganic materials. In this work, the effect of the ILs anion type on morphology and photoactivity of TiO2 is presented. The preparation of TiO2 microparticles with spherical structure was successfully achieved by solvothermal method, using tetra-tert-butyl orthotitatane (TBOT) as the precursor. The reaction process was assisted by an ionic liquids 1-butyl-3-methylimidazolium bromide [BMIM][Br], 1-butyl-3-methylimidazolium tetrafluoroborate [BMIM][BF4] and 1-butyl-3-methylimidazolium haxafluorophosphate [BMIM][PF6]. Various molar ratios of all ILs to TBOT (IL:TBOT) were chosen. For comparison, reference TiO2 was prepared using the same method without IL addition. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Brenauer-Emmett-Teller surface area (BET), NCHS analysis, and FTIR spectroscopy were used to characterize the surface properties of the samples. The photocatalytic activity was investigated by means of phenol photodegradation in the aqueous phase as a model pollutant, as well as formation of hydroxyl radicals based on detection of fluorescent product of coumarine hydroxylation. The analysis results showed that the TiO2 microspheres had spherical structure with the diameters ranging from 1 to 6 µm. The TEM micrographs gave a bright observation of the samples in which the particles were comprised of inter-aggregated crystals. It could be also observed that the IL-assisted TiO2 microspheres are not hollow, which provides additional information about possible formation mechanism. Application of the ILs results in rise of the photocatalytic activity as well as BET surface area of TiO2 as compared to pure TiO2. The results of the formation of 7-hydroxycoumarin indicated that the increased amount of ·OH produced at the surface of excited TiO2 for samples TiO2_ILs well correlated with more efficient degradation of phenol. NCHS analysis showed that ionic liquids remained on the TiO2 surface confirming structure directing role of that compounds.

Keywords: heterogeneous photocatalysis, IL-assisted synthesis, ionic liquids, TiO2

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Authors: Amira M. Hassanein, Nehal A. Salahuddin, Atsunori Matsuda, Toshiaki Hattori, Mona N. Elfiky

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New insights into the design of highly sensitive, carbon-based electrochemical sensors are presented in this work. This was achieved by exploring the interesting properties of conductive (Mg/Al) layered double hydroxide- Dodecyl Sulphate/Polypyrrole nanocomposites which were synthesized by in-situ polymerization of pyrrole during the assembly of (Mg/Al) layered double hydroxide, and by employing the anionic surfactant Dodecyl sulphate as a modifier. The morphology and surface area of the nanocomposites changed with the percentage of Pyrrole. Under optimal conditions, the modified carbon paste electrode successfully achieved detection limits of 0.057 and 0.134 nmol.L-1 of Terazosin hydrochloride in pharmaceutical formulation and spiked human serum fluid, respectively. Moreover, the sensors are highly stable, reusable, and free from interference by other commonly present excipients in drug formulations.

Keywords: layered double hydroxide, polypyrrole, terazosin hydrochloride, square-wave adsorptive anodic stripping voltammetry

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Authors: Rita Riekstina-Dolge, Zanda Kruma, Fredijs Dimins, Inta Krasnova, Daina Karklina

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Apple juice is the main raw material for cider production. In this study apple juices obtained from 14 dessert and crab apples grown in Latvia were investigated. For all samples total phenolic compounds, tannins and individual phenolic compounds content were determined. The total phenolic content of different variety apple juices ranged from 650mg L-1 to 4265mg L-1. Chlorogenic acid is the predominant phenolic compound in all juice samples and ranged from 143.99mg L-1 in ‘Quaker Beauty’ apple juice to 617.66mg L-1 in ‘Kerr’ juice. Some dessert and crab apple juices have similar phenolic composition, but in several varieties such as ‘Cornelie’, ‘Hyslop’ and ‘Riku’ it was significantly higher. For cider production it is better to blend different kinds of apple juices including apples rich in high phenol content ('Rick', 'Cornelie') and also, for successful fermentation, apples rich in sugars and soluble solids content should be used in blends.

Keywords: apple juice, phenolic compounds, hierarchical cluster analysis, cider production

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Authors: Francesca Crivellari, Nicholas Mavrogiannis, Zachary Gagnon

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Portable diagnostic methods have become increasingly important for a number of different purposes: point-of-care screening in developing nations, environmental contamination studies, bio/chemical warfare agent detection, and end-user use for commercial health monitoring. The cheapest and most portable methods currently available are paper-based – lateral flow and dipstick methods are widely available in drug stores for use in pregnancy detection and blood glucose monitoring. These tests are successful because they are cheap to produce, easy to use, and require minimally invasive sampling. While adequate for their intended uses, in the realm of blood-borne pathogens and numerous cancers, these paper-based methods become unreliable, as they lack the nM/pM sensitivity currently achieved by clinical diagnostic methods. Clinical diagnostics, however, utilize techniques involving surface plasmon resonance (SPR) and enzyme-linked immunosorbent assays (ELISAs), which are expensive and unfeasible in terms of portability. To develop a better, competitive biosensor, we must reduce the cost of one, or increase the sensitivity of the other. Electric fields are commonly utilized in microfluidic devices to manipulate particles, biomolecules, and cells. Applications in this area, however, are primarily limited to interfaces formed between immiscible interfaces. Miscible, liquid-liquid interfaces are common in microfluidic devices, and are easily reproduced with simple geometries. Here, we demonstrate the use of electrical fields at liquid-liquid electrical interfaces, known as fluidic dielectrophoresis, (fDEP) for biodetection in a microfluidic device. In this work, we apply an AC electric field across concurrent laminar streams with differing conductivities and permittivities to polarize the interface and induce a discernible, near-immediate, frequency-dependent interfacial tilt. We design this aqueous electrical interface, which becomes the biosensing “substrate,” to be intelligent – it “moves” only when a target of interest is present. This motion requires neither labels nor expensive electrical equipment, so the biosensor is inexpensive and portable, yet still capable of sensitive detection. Nanoparticles, due to their high surface-area-to-volume ratio, are often incorporated to enhance detection capabilities of schemes like SPR and fluorimetric assays. Most studies currently investigate binding at an immobilized solid-liquid or solid-gas interface, where particles are adsorbed onto a planar surface, functionalized with a receptor to create a reactive substrate, and subsequently flushed with a fluid or gas with the relevant analyte. These typically involve many preparation and rinsing steps, and are susceptible to surface fouling. Our microfluidic device is continuously flowing and renewing the “substrate,” and is thus not subject to fouling. In this work, we demonstrate the ability to electrokinetically detect biomolecules binding to functionalized nanoparticles at liquid-liquid interfaces using fDEP. In biotin-streptavidin experiments, we report binding detection limits on the order of 1-10 pM, without amplifying signals or concentrating samples. We also demonstrate the ability to detect this interfacial motion, and thus the presence of binding, using impedance spectroscopy, allowing this scheme to become non-optical, in addition to being label-free.

Keywords: biodetection, dielectrophoresis, microfluidics, nanoparticles

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Authors: Narjes Dashti, Nida Ali, Magdy Montasser, Vineetha Cherian

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The influence of two PGPR strains, Pseudomonas aeruginosa and Stenotrophomonas rhizophilia, on fruit yields, pomological traits and chemical contents of tomato (Solanum lycopersicum) fruits were studied. The study was conducted separately on two different cultivar varieties of tomato, namely Supermarmande and UC82B. The results indicated that the presence of the PGPR almost doubled the average yield per plant. There was a significant improvement in the pomological qualities of the PGPR treated tomato fruits compared to the corresponding healthy treatments especially in traits such as the average fruit weight, height, and fruit volume. The chemical analysis of tomato fruits revealed that the presence of the PGPRs increased the total protein, lycopene, alkalinity and phenol content of the tomato fruits compared to the healthy controls. They had no influence on the reduced sugar, total soluble solids or the titerable acid content of fruits. However their presence reduced the amount of ascorbic acid in tomato fruits compared to the healthy controls.

Keywords: PGPR, tomato, fruit quality

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Authors: Hawraa Zbeeb, Hala Khalifeh, Mohamad Khalil, Francesca Storace, Francesca Baldini, Giulio Lupidi, Laura Vergani

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Sarcopoterium spinosum, a widely distributed spiny shrub belonging to the Rosaceae family, is rich in essential and beneficial constituents. In fact, S. spinosum fruits and roots are traditionally used as herbal medicine in the eastern Mediterranean landscape, and this shrub is mentioned as a medicinal plant in a large number of ethnobotanical surveys. Aqueous root extracts from S. spinosum are used by traditional medicinal practitioners for weight loss treatment of diabetes and pain. Moreover, the anti-diabetic activity of S. spinosum root extract has been reported in different studies, but the beneficial effects of aerial parts, especially fruits, have not been elucidated yet. The aim of the present study was to investigate the in vitro antioxidant and lipid-lowering properties of an ethanolic extract from S. spinosum fruits using both hepatic (FaO) and endothelial (HECV) cells in an attempt to evaluate its possible employment as a nutraceutical supplement. First of all, in vitro spectrophotometric assays were employed to characterize the extract. The total phenol content (TPC) was evaluated by Folin–Ciocalteu spectrophotometric method and the radical scavenging activity was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. After that, the beneficial effects of the extract were tested on cells. FaO cells treated for 3 hours with 0.75 mM oleate/palmitate mix (1:2 molar ratio) mimic in vitro a moderate hepato-steatosis. HECV cells exposed for 1 hour to 100 µM H₂O₂ mimic an oxidative insult leading to oxidative stress conditions. After the metabolic and oxidative insult, both cell lines were treated with increasing concentrations of the S. spinosum extract (1, 10, 25 µg/mL) for 24 hours. The results showed the S. spinosum ethanolic extract is rather rich in phenols (TPC of 18.6 mgGAE/g dry extracts). Moreover, the extract showed a good scavenging ability in vitro (IC₅₀ 15.9 µg/ml and 10.9 µg/ml measured by DPPH and ABTS assays, respectively). When the extract was tested on cells, the results showed that it could ameliorate some markers of cell dysfunction. The three concentrations of the extract led to a significant decrease in the intracellular triglyceride (TG) content in steatotic FaO cells measured by spectrophotometric assay. On the other hand, HECV cells treated with increasing concentrations of the extract did not result in a significant decrease in both lipid peroxidation measured by the Thiobarbituric Acid Reactive Substances (TBARS) assay, and in reactive oxygen species (ROS) production measured by fluorometric analysis after DCF staining. Interestingly, the ethanolic extract was able to accelerate the wound repair of confluent HECV cells with respect to H₂O₂-insulted cells as measured by T-scratch assay. Taken together, these results seem to indicate that the ethanol extract from S. spinosum fruits is rich in phenol compounds and plays considerable lipid-lowering activity in vitro on steatotic hepatocytes and accelerates wound healing repair on endothelial cells. In light of that, the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical applications.

Keywords: antioxidant activity, ethanolic extract, lipid-lowering activity, phenolic compounds, Sarcopoterium spinosum fruits

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Authors: Sylvain Caillol

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Uncertainty in terms of price and availability of petroleum, in addition to global political and institutional tendencies toward the principles of sustainable development, urge chemical industry to a sustainable chemistry and particularly the use of renewable resources in order to synthesize biobased chemicals and products. We propose a platform approach for the synthesis of various building blocks from cardanol in one or two-steps syntheses. Cardanol, which is a natural phenol, is issued from Cashew Nutshell Liquid (CNSL), a non-edible renewable resource, co-produced from cashew industry in large commercial volumes. Cardanol is particularly interesting to replace fossil aromatic groups in polymers and materials. Our team studied various routes for the synthesis of cardanol-derived biobased building blocks used after that in polymer syntheses. For example, we used phenolation to dimerize/oligomerize cardanol to propose increase functionality of cardanol. Thio-ene was used to synthesize new reactive amines. Epoxidation and (meth)acrylation were also used to insert oxirane or (meth)acrylate groups in order to synthesize polymers and materials.

Keywords: cardanol, cashew nutshell liquid, epoxy, vinyl ester, latex, emulsion

Procedia PDF Downloads 176

Authors: B. Voucko, M. Benkovic, N. Cukelj, S. Drakula, D. Novotni, S. Balbino, D. Curic

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Oxidative stress is considered as one of the causes leading to metabolic disorders in humans. Therefore, the ability of antioxidants to inhibit free radical production is their primary role in the human organism. Antioxidants originating from cereals, especially flavonoids and polyphenols, are mostly bound and indigestible. Micronization damages the cell wall which consecutively results in bioactive material to be more accessible in vivo. In order to ensure complete fragmentation, micronization is often combined with high temperatures (e.g., for bran 200°C) which can lead to degradation of bioactive compounds. The innovative non-thermal technology of cryo-milling is an ultra-fine micronization method that uses liquid nitrogen (LN2) at a temperature of 195°C to freeze and cool the sample during milling. Freezing at such low temperatures causes the material to become brittle which ensures the generation of fine particles while preserving the bioactive content of the material. The aim of this research was to determine if production of ultra-fine material with cryo-milling will result in the augmentation of available bioactive compounds of buckwheat and pumpkin seed cake. For that reason, buckwheat and pumpkin seed cake were ground in a ball mill (CryoMill, Retch, Germany) with and without the use of LN2 for 8 minutes, in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm) at an oscillation frequency of 30 Hz. The cryo-milled samples were cooled with LN2 for 2 minutes prior to milling, followed by the first cycle of milling (4 minutes), intermediary cooling (2 minutes), and finally the second cycle of milling (further 4 minutes). A continuous process of milling was applied to the samples ground without freezing with LN2. Particle size distribution was determined using the Scirocco 2000 dry dispersion unit (Malvern Instruments, UK). Antioxidant activity was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) test and ferric reducing antioxidant power (FRAP) assay, while the total phenol content was determined using the Folin Ciocalteu method, using the ultraviolet-visible spectrophotometer (Specord 50 Plus, Germany). The content of the free phenolic acids, rutin in buckwheat, tyrosol in pumpkin seed cake, was determined with an HPLC-PDA method (Agilent 1200 series, Germany). Cryo-milling resulted in 11 times smaller size of buckwheat particles, and 3 times smaller size of pumpkin seed particles than milling without the use of LN2, but also, a lower uniformity of the particle size distribution. Lack of freezing during milling of pumpkin seed cake caused a formation of agglomerates due to its high-fat content (21 %). Cryo-milling caused augmentation of buckwheat flour antioxidant activity measured by DPPH test (23,9%) and an increase in available rutin content (14,5%). Also, it resulted in an augmentation of the total phenol content (36,9%) and available tyrosol content (12,5%) of pumpkin seed cake. Antioxidant activity measured with the FRAP test, as well as the content of phenolic acids remained unchanged independent of the milling process. The results of this study showed the potential of cryo-milling for complete raw material utilization in the food industry, as well as a tool for extraction of aimed bioactive components.

Keywords: bioactive, ball-mill, buckwheat, cryo-milling, pumpkin seed cake

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Authors: Muhammad Ali Syed, Arshad Saleem Bhatti, Chen-zhong Li, Habib Ali Bokhari

Abstract:

Surface plasmon resonance (SPR) biosensors have emerged as a promising technique for bioanalysis as well as microbial detection and identification. Real time, sensitive, cost effective, and label free detection of biomolecules from complex samples is required for early and accurate diagnosis of infectious diseases. Like many other types of optical techniques, SPR biosensors may also be successfully utilized for microbial detection for accurate, point of care, and rapid results. In the present study, we have utilized a commercially available automated SPR biosensor of BI company to study the microbial detection form water samples spiked with different concentration of Staphylococcus aureus bacterial cells. The gold thin film sensor surface was functionalized to react with proteins such as protein G, which was used for directed immobilization of monoclonal antibodies against Staphylococcus aureus. The results of our work reveal that this immunosensor can be used to detect very small number of bacterial cells with higher sensitivity and specificity. In our case 10^3 cells/ml of water have been successfully detected. Therefore, it may be concluded that this technique has a strong potential to be used in microbial detection and identification.

Keywords: surface plasmon resonance (SPR), Staphylococcus aureus, biosensors, microbial detection

Procedia PDF Downloads 475

Authors: Abdallah Habbab, Khaled Sekkoum, Nasser Belboukhari

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Objective: This study is designed to examine the chemical composition, antioxidant and antibacterial activities of the essential oil extracted from leaves of Vitex agnus-castus. Methods: The essential oils of dry leaves of Vitex agnus-castus L. were obtained by hydro-distillation, afforded oil in the yield of 5.5% and their volatile constituents were identified by GC/MS. Antioxidant activity of the sample was determined by test system DPPH. Antifungal activity was tested against three fungal strains (Aspergillus flavus, Penicillium escpansum and Aspergillus ochraceus) by direct contact method. Results: Forty-three compounds were identified, representing 98.02% of the oil. Major components of the oil were 1,8-cineole (18.27 %), caryophyllene (8.60 %), N-(M-fluorophenyl)-maleimide (6.30 %), (+)-epi-bicyclosesquiphellandrene (6.00 %), terpinen-4-ol (5.57 %), pyrrolo (3,2,1-jk) carbazole (5.43 %), caryophyllene oxide (4.79 %), and phenol (4.09 %). Conclusion: The chemical constituents in the essential oil from the locally grown Vitex agnus-castus were identified. Therefore, the essential oil of Vitex agnus-castus is an active candidate which would be used as antioxidant, or antifungal agent in new drugs preparation for therapy of diseases.

Keywords: Vitex agnus-castus, essential oil, GC/MS, DPPH, 1, 8-cineole

Procedia PDF Downloads 463

Authors: Tian-Fang Kang, Xue Sun

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DNA damage induced by tetrachloro-1,4-benzoquinone (TCBQ), a reactive metabolite of pentachloro-phenol (PCP), was investigated using a glassy carbon electrode (GCE) modified with calf thymus double-stranded DNA (ds-DNA) in this work. DNA modified films were constructed by layer-by-layer adsorption of polycationic poly(diallyldimethyl- ammonium chloride) (PDDA) and negatively charged ds-DNA on the surface of a glassy carbon electrode. The DNA intercalator [Ru(bpy)2(dppz)]2+ (bpy=2, 2′-bipyridine, dppz0dipyrido [3, 2-a: 2′,3′-c] phenazine) was chosen as an electrochemical probe to detect DNA damage. After the sensor was incubated in 0.1 M pH 7.3 phosphate buffer solution (PBS) for 30min, the intact PDDA/DNA film produced a sensitive electrochemiluminescent (ECL) signal. However, after the sensor was incubated in 100 μM TCBQ or a mixed solution of 100 μM TCBQ and 2 mM H2O2, ECL signal decreased significantly. During the incubation of DNA in TCBQ or TCBQ-H2O2 solution, the double-helix of DNA was damaged, which resulted in the decrease of Ru-dppz bound to DNA. Additionally, the results were verified independently by fluorescence experiments. This paper provides a sensitive method to directly screen DNA damage induced by chemicals in the environment.

Keywords: DNA damage, detection, electrochemiluminescence, sensor

Procedia PDF Downloads 410

Authors: Alireza Vakili, Shahab Ehtesham, Mohsen Danesh Mesgaran, Mahdi Paktinat

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The goal of this study is to determine the chemical compounds of brown Iranian propolis(BIP) extracts and to show flavonoids and phenol effects on nitrogen ammonia (NH3-N) in in vitro. Experimental samples were including two diets with different concentrate: forage ratio (80:20 and 60:40) with eight treatments (1:Control diet 60:40 without BIP,2: 60:40 diet with 25% BIP, 3:60:40 diet with 50% BIP, 4: 60:40 diet with 75% BIP,5: Control diet 80:20 without BIP,6: 80:20 diet with 25% BIP,7: 80:20 diet with 50% BIP and 8: 80:20 diet with 75% BIP) and eight repeats. The trial was analyzed considering a completely randomized design by the GLM procedure of SAS 9.1. Means among treatment were compared by Tukey test. The results of this study showed that in food with 80:20 (concentrate: forage), adding BIP 25% did not statistically change NH3-N (p > 0.05) compared to the control treatment but there was a significant difference (p < 0.05) between the effect of BIP 50% on NH3-N compared to the BIP 25% and the control. In diet with 60:40 (concentrate: forage), there was no significant difference between the effect of BIP 25% on NH3-N and the control, nor was there a significant difference between the effect of BIP 50% and 75%, while a significant difference (p < 0.05) between BIP 50% and 75% and the rest was observed. The propolis extract makes nitrogen ammonia decrease. This may help the nitrogen retain longer in ruminants.

Keywords: brown Iranian propolis, in vitro, nitrogen ammonia, ruminant

Procedia PDF Downloads 488

Authors: Mian Gul Sayed, Rahim Shah, Fazal Mabood, Najeeb Ur Rahman, Maher Noor

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Embarking on the frontier of molecular advancement, this study focuses on the synthesis and characterization of a distinct class of porphyrin derivatives—specifically, the 5, 10, 15, 20-tetrakis (3-bromopropoxyphenyl) porphyrins. Through meticulous synthetic methodologies, these derivatives are crafted, strategically incorporating bromopropoxyphenyl moieties at distinct positions within the porphyrin framework. This research aims to unravel the structural intricacies and explore the potential applications of these compounds through a detailed characterization utilizing advanced analytical techniques. 5, 10, 15, 20, tetrakis (4-hydroxyphenyl) porphyrin was synthesized by treating pyrrole and p- hydroxylbenzaldehyde. 5, 10, 15, 20, tetrakis-(4-hydroxyphenyl) was converted into 5, 10, 15, 20, tetrakis (4-bromoalkoxyphenyl) porphyrin. 5,10,15, 20-Tetrakis -(4-bromoalkoxyphenyl) porphyrin was treated with Isopropyl phenol, para-Aminophenol, hydroquinone, 2-Naphthol, 1-Naphthol and Hydroquinone and different derivatives of ether-linked were obtained. The synthesized compounds were analyzed using contemporary spectroscopic techniques like UV-Vis, NMR and Mass spectrometry. The synthesized compounds were also tested for their biological activities like antioxidants and anti-inflammatory.

Keywords: tetraphenyl porphyrin, NMR, antioxidant, anti-inflammatory

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Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

Procedia PDF Downloads 136