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Nanomaterials have attracted specific consideration due to their specific characteristics and uses in several promising fields. In the present study, Chondrilla juncea was employed as a biological extract to facilitate the reduction of copper and silver ions within garlic peel powders. The resulting garlic-CuO and garlic-AgO nanocomposites were characterized using several analytical methods including FTIR, TGA/DTG, SEM, TEM, and XRD analyses. The garlic peel exhibited a rough surface. The nanoparticles were evenly dispersed across its surface. The incorporation of CuO and AgO nanoparticles affected the crystal structure of garlic peel. The establishment of CuO and AgO nanoparticles was evidenced by the highest residual mass values observed for the prepared nanocomposites. The thermogravimetric analysis showed that the prepared nanocomposites had lower thermal stability compared with garlic peel powders. The prepared nanocomposites were used for catalytic degradation of naphthol blue black B and calmagite. The decolorization process depended on the quantity of H2O2, initial concentration of azo dyes, duration of contact, and temperature of the bath. The calculated activation energy (Ea) values for the garlic-CuO nanocomposites were found to be 18.44 kJ mol-1 and 23.28 kJ mol-1 for calmagite and naphthol solutions, respectively. However, those calculated for garlic-AgO nanocomposites were found to be 50.01 kJ mol-1 and 12.44 kJ mol-1 for calmagite and naphthol, respectively.

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Prorocentrum lima, a widely distributed dinoflagellate known for its production of harmful biotoxins, poses a significant threat to humans, aquaculture, and marine ecosystems. As a result, the detection of this toxic alga in coastal waters has become an urgent research focus. In this study, a rapid, sensitive, and cost-effective detection method based on loop-mediated isothermal amplification (LAMP) was developed to identify P. lima. In this method, cell extracts of P. lima were diluted and used directly as templates for amplification, eliminating the need for nucleic acid purification and simplifying the detection process. Hydroxy naphthol blue (HNB) was incorporated into the reaction mix to facilitate result interpretation, enabling visual determination of the amplification outcome with the naked eye. The entire detection process, from DNA extraction to template amplification and product detection, could be completed within 80 min using a simple constant temperature-control device. This LAMP-based detection method demonstrated excellent reliability, specificity, and a low detection limit of 5.87 cells/mL for DNA crude extract. The assay offered an efficient alternative to PCR for rapid detection of P. lima. By streamlining the detection process and offering a visual readout, this technique holds promise for efficient and routine monitoring of harmful algal species, benefitting both research efforts and environmental management strategies.

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In recent years, a new type of Spiroplasma has been found that can cause "tremor disease" of the Chinese mitten crab Eriocheir sinensis. The outbreak of epidemic tremor disease has caused a serious setback in the Chinese mitten crab farming industry, with an incidence rate of more than 30% and mortality rates of 80-100%. Therefore, finding a sensitive method to detect tremor disease in E. sinensis has become a current research focus. In this research, a loop-mediated isothermal amplification detection method coupled with hydroxynaphthol blue dye (LAMP-HNB) was developed and used to rapidly detect Spiroplasma eriocheiris. First, we designed and synthesized specific outer primers, inner primers and loop primers based on the 16S ribosomal RNA gene of S. eriocheiris. Second, the LAMP-HNB detection method for S. eriocheiris was successfully established by screening the primers, adjusting the temperature and time of the reaction, and optimizing the concentrations of Mg2+ and dNTPs. In the specific tests, only samples infected with S. eriocheiris showed positive results, and other infections caused by bacteria and parasites tested negative, proving that the test has high specificity. Moreover, the detection limit was 2.5 × 10-6 ng/µL, indicating high sensitivity. This method for detecting S. eriocheiris provides rapid visual output based on LAMP-HNB detection and is a simple, fast, sensitive, and inexpensive method that can be applied to a wide range of field investigations.

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Patchouli (Pogostemon cablin), a highly valued medicinal plant, suffers significant economic losses following infection with Broad bean wilt virus 2 (BBWV-2) and Peanut stripe virus (PStV). In this study, a field-based isothermal technique called reverse transcription loop-mediated isothermal amplification (RT-LAMP) was established for an early and specific detection of BBWV-2 and PStV. The oligo primers were designed to target the coat protein genes of PStV and BBWV-2. The reaction conditions, such as temperature and time duration, were optimized to 65 °C for 60 min. The LAMP amplicons positive for PStV and BBWV-2 revealed characteristic ladder-type bands following agarose gel electrophoresis. Further, a colorimetric assay using a metal ion-based indicator (Hydroxy-naphthol blue, HNB) was conducted to visualize the amplified products with the naked eye, thus facilitating accessibility to field practices. The assay developed in this study was found to be virus specific, and was 100 times more sensitive than RT-PCR. Thus, the RT-LAMP assay established in this study is quick, reliable, and cost-effective for the accurate identification of BBWV-2 and PStV. It will facilitate the screening of patchouli planting materials.  Further, it may reduce the risk of virus spread and could be helpful in phytosanitary programs.

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This work provides a first-time comparative study examining the photocatalytic activity of functionalized TiO2-based composites to eliminate naphthol blue in Batik wastewater. Reduced graphene oxide (RGO) was synthesized by oxidizing solid graphite using the Hummers' method followed by sonication and reduction. N-doped TiO2 (N-TiO2) was synthesized from titanium tetrachloride (TiCl4) and urea (CH4N2O) precursors by the sol-gel method. N-TiO2 modified RGO (RGO/NT) was synthesized using a hydrothermal method from N-TiO2 and RGO. Prepared TiO2-based composites and commercial TiO2, for comparison were characterized using Fourier transform infrared spectrometer (FTIR), X-Ray diffractometer (XRD), scanning electron microscope-energy dispersive X-ray (SEM-EDX), and UV-Vis diffuse reflectance spectrometer (UV-Vis DRS). FTIR characterization indicated Ti-N bonding in N-TiO2 and RGO/NT. XRD patterns showed that commercial TiO2 had a rutile phase, while N-TiO2 and RGO/NT had an anatase phase with crystal sizes of 30.09, 16.28, and 12.02 nm, respectively. SEM results displayed the presence of small and glossy white N-TiO2 dispersed on the surface of RGO. Characterization using UV-Vis DRS showed that the band gap energy values for TiO2, N-TiO2, and RGO/NT were 3.25, 3.12, and 3.08 eV with absorption regions at the wavelengths of 382, 398, and 403 nm, respectively. The highest photocatalytic activity for RGO/NT for degrading naphthol blue was obtained at pH 5, with a photocatalyst mass of 60 mg, and an irradiation of 15 min. Photocatalytic degradation by RGO/NT on Batik wastewater under visible light showed higher effectivity than under UV light.

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In this study, activated carbon produced from oak cupules (ACOC) was prepared using chemical activation with H3PO4. ACOC is subsequently used as an adsorbent to facilitate the removal of an acidic dye, naphthol blue black (NBB), and basic dye crystal violet (CV) from aqueous solutions. The ACOC was characterized by FTIR spectroscopy, XRD, and SEM. The adsorption isotherm data fits well with the Langmuir model for NBB and CV. The kinetic models of adsorption of NBB and CV by ACOC were pseudo-first order and pseudo-second order, respectively. Thermodynamic parameters were evaluated and indicated that the adsorption of both dyes onto ACOC was endothermic and spontaneous. The adsorption capacity of ACOC reached 208 mg g-1 for NBB and 658 mg g-1 for CV. ACOC was shown to be a promising adsorbent for the removal of NBB and CV from aqueous solutions.

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Histological methods allow to characterize the developmental changes that occur in plant tissues during in vitro culturing of explants. Different procedures to assess these changes have been reported in the literature. Here we describe a simple histological procedure to evaluate the development of explants using the fixation with paraformaldehyde, embedding with methacrylate resin and staining with periodic acid Schiff's reagent and naphthol blue black. This method allows to obtain high contrast in the stained cells, for a better characterization of cellular events that occur during coconut somatic embryogenesis.

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Chilli leaf curl disease caused by whitefly transmitted begomoviruses is an important constraint to chilli (Capsicum anuum L.) cultivation in India. Tomato leaf curl Joydebpur virus (ToLCJoV) was characterized and identified as incitant of leaf curl disease through rolling circle amplification (RCA) and PCR assay from the symptomatic samples collected from Uttar Pradesh, India. Although PCR assay provides the gold standard in diagnostics, this method consumes more time and requires convenient portable instruments. Therefore, a loop-mediated isothermal amplification (LAMP) assay was developed for the detection of ToLCJoV by targeting the AC1 and AC2 region. Detection has been achieved through a laddered pattern of amplification in agarose gel electrophoresis. The assay has detected ToLCJoV in a total DNA concentration of 1 × 10-1 ng indicating 200-fold higher sensitivity than that of the PCR. Further, the replacement of total DNA with leaf extracts using the grinding buffer and GES buffer coupled with LAMP assay also detected the presence of ToLCJoV in the infected chilli samples. With this assay, ToLCJoV can be detected in less than 2 h without DNA extraction. Besides, this assay will be highly useful in discriminating the leaf curl disease etiology by ToLCJoV from other begomoviruses and insects (thrips and mites). To the best of our knowledge, this is the first report of a LAMP assay for the detection of ToLCJoV.

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Staphylococcus aureus is a common clinical bacterial pathogen that can cause a diverse range of infections. The establishment of a rapid and reliable assay for the early diagnosis and detection of S. aureus is of great significance. In this study, we developed a closed-tube loop-mediated isothermal amplification (LAMP) assay for the visual detection of S. aureus using the colorimetric indicator hydroxy naphthol blue (HNB). The LAMP reaction was optimized by adjusting the amplification temperature, the concentrations of Mg2+, dNTP, and HNB, and the incubation time. In the optimized reaction system, the specificity of LAMP for S. aureus was 100%. The results established that this method accurately identified S. aureus, with no cross-reactivity with 14 non-S. aureus strains. The limit of detection (LOD) of LAMP was 8 copies/reaction of purified plasmid DNA or 400 colony-forming units/reaction of S. aureus. Compared with conventional PCR, LAMP lowered the LOD by tenfold. Finally, 220 clinically isolated strains of S. aureus and 149 non-S. aureus strains were used to evaluate the diagnostic efficacy of LAMP (test accuracy, 99.46%). The findings indicated that LAMP is a reliable test for S. aureus and could be a promising tool for the rapid diagnosis of S. aureus infections.

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Hepatopancreatic microsporidiosis (HPM) is an infectious shrimp disease caused by the microsporidian Enterocytozoon hepatopenaei (EHP). In recent years, the widespread occurrence of EHP poses a significant challenge to the shrimp aquaculture industry. Early, rapid and accurate diagnosis of EHP infection is very much essential for the control of HPM crop-related losses. Loop-mediated isothermal amplification (LAMP) is a robust, sensitive, cost-effective disease diagnostic technique. Here, we demonstrate an improved, simple, closed-tube, colorimetric EHP LAMP diagnostic assay. LAMP assay was illustrated with the specific EHP spore wall protein (SWP) gene primers. Naked eye visual detection of LAMP amplicons was achieved using Hydroxy naphthol blue (HNB) or Phenol red dye without opening the tubes. This LAMP assay is efficient in detecting the EHP pathogen in all clinical samples include shrimp hepatopancreas, FTA card samples, feces, pond water, and soil. Also, the elution of EHP DNA from FTA cards was demonstrated within 17 min using a simple dry bath. In clinical evaluation, the visual LAMP assay established 100% diagnostic sensitivity and 100% diagnostic specificity. The visual LAMP assay is rapid, can detect the EHP pathogen within 40 min using a simple dry bath, and does not require any expensive instruments and technical proficiency. In conclusion, this visual LAMP protocol is a user-friendly, specific assay that can be conceivably operated at the farm-site/ resource-limited settings by the farmer himself with simple equipment.

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In this study, copper oxide nanoparticles, prepared using pistacia vera hull extract, were immobilized onto chitosan. FT-IR spectrum of copper oxide-chitosan exhibited chemical shifting of the main peaks of the biopolymer indicating that hydroxyl and amino groups were reacted with copper oxide nanoparticles. SEM features showed spherical surface and physical stability of the composite. The shifting of the burning temperature in DTA from 278.5 °C to 212.6 °C in the composite proved the interaction between chitosan and copper oxide nanoparticles. The composite was applied for the oxidative degradation of naphthol blue black in the presence of H2O2. The change of copper oxide nanoparticles loading, time, dye concentration, temperature, and oxidant dose were studied. The degradation yield reached 86 % (C0 = 30 mg/L, T=20 °C, H2O2 = 8 mL/L). The activation energy (Ea), entropy (ΔS*) and enthalpy (ΔH*) were equal to 45.558 KJ. mol-1, -116.203 J mol-1 K-1 and 42.986 kJ mol-1, respectively.

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Lab-on-a-disc (LOD) has emerged as a promising candidate for a point-of-care testing (POCT) device because it can effectively integrate complex fluid manipulation steps using multiple layers of polymeric substrates. However, it is still highly challenging to design and fabricate temperature measurement and heating system in non-contact with the surface of LOD, which is a prerequisite to successful realization of DNA amplification especially with a rotatable disc. This study presents a Lab-on-a-disc (LOD)-based automatic loop-mediated isothermal amplification (LAMP) system, where a thermochromic coating (<~420 µm) was used to distantly measure the chamber's temperature and a micro graphite film was integrated into the chamber to remotely absorb laser beam with super high efficiency. We used a deep learning network to more consistently analyze the product of LAMP than we could with the naked eye. Consequently, both temperature heating and measurement were carried out without a physical contact with the surface of LOD. The experimental results show that the proposed approach, which no previous work has attempted, was highly effective in realizing LAMP in LOD.

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Invasive Aedes mosquito species (Diptera: Culicidae) are of public health concern in Europe because they are either recognized or potential vectors of pathogens. Loop-mediated isothermal amplification (LAMP) is a rapid and simple method for amplifying DNA with high specificity and efficiency, with the technique having potential for application in the field, including in high-throughput format. Specific LAMP assays based on rDNA internal transcribed spacers 1 or 2 sequences, considering intraspecies variability at these loci, were developed for Aedes aegypti, Aedes albopictus, Aedes japonicus, Aedes koreicus and the indigenous Aedes geniculatus. No such assays could be developed for Aedes atropalpus and Aedes triseriatus because both loci were too short to serve as target. The assays rely on the clearly visible colour change from violet to sky blue after successful amplification. Sensitivity of egg detection was confirmed with ratios of up to one mosquito egg in 99 other eggs. Simple sample preparation of adults or eggs by mechanical homogenization in water required an additional heat treatment or centrifugation step to avoid non-specific colour changes. Thus, further technical improvements are needed to render these assays truly field-applicable, which would greatly facilitate surveillance of these invasive mosquito species and allow for prompt implementation of control measures.

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In this study, ultrasound-assisted cloud point extraction (UA-CPE) method was developed for the determination of Ni(II) and Co(II) in milk-based products. After extraction and preconcentration, the Ni(II) and Co(II) contents of samples were determined by flame atomic absorption spectrometry (FAAS). After their complexation with hydroxy naphthol blue (HNB) in the presence of cationic surfactant, CTAB at pH 4.0, the Ni(II) and Co(II) were taken within the micellar phase of nonionic surfactant, TX-114. The micellar phase containing the analytes were diluted to a volume of 0.7 mL with 1.0-mol/L HNO3 in ethanol to reduce its viscosity and to facilitate sample treatment and then was analyzed by FAAS. The various analytical parameters affecting UA-CPE efficiency were investigated. The analytical features obtained after optimization are as follows: limits of detection are 0.56 and 0.78 µg/L; sensitivity enhancement factors are 48.6 and 53.9; the calibration curves were linear 3-180 and 2-160 µg/L for Co(II) and Ni(II), respectively, after preconcentration of 50-fold. The precision (as RSD%) between 1.8-3.6% and 2.2-3.8% (25 and 100 µg/L, n = 5) for Ni(II) and Co(II), respectively. The accuracy was statistically verified by analysis of two certified reference material samples (CRMs), including recovery studies after spiking. The method was applied to the analysis of milk-based samples with satisfied results.

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In this study, mineralization and color removal performance of electro-Fenton method were examined in water containing naphthol blue black (NBB), a diazo dye. NBB was totally converted to intermediate species in a 15-min electrolysis at 60 mA, but complete de-colorization took 180 min. A very high oxidation rate constant ((3.35 ±â€¯0.21) x 1010 M-1s-1) was obtained for NBB, showing its high reactivity towards hydroxyl radicals. A very high total organic carbon (TOC) removal value (45.23 mg L-1) was obtained in the first 60 min of the electro-Fenton treatment of an aqueous solution of NBB (0.25 mM) at 300 mA, indicating the mineralization efficiency of the electro-Fenton method. Mineralization current efficiency values obtained at 300 mA gradually decreased from 24.18% to 4.47% with the electrolysis time, indicating the presence of highly parasitic reactions. Gas chromatography-mass spectrometry analyses revealed that the cleavage of azo bonds of NBB led to formation of different aromatic and aliphatic oxidation intermediates. Ion chromatography analysis showed that ammonium, nitrate and sulfate were the mineralization end-products. The concentration of sulfate ion reached to its quantitative value at the 4th h of electrolysis. On the other hand, the total concentration of ammonium and nitrate ions reached to only 61% of the stoichiometric amount of initial nitrogen after a 7 h electrolysis. Finally, it can be said that the electro-Fenton method is a suitable and efficient method for the removal of NBB and its intermediates from water.

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This work explores the effect of persulfate (PS) on the sonochemical degradation of organic pollutants taking naphthol blue black (NBB), an anionic diazo dye, as a substrate model. The sonolytic experiments were conducted in the absence and presence of PS under various experimental conditions including acoustic power (10-80W), frequency (20 and 585kHz) and saturating gas (argon, air and nitrogen). Experimental results showed that PS decomposition into sulfate radical (SO4-) takes place by sonolysis and increasing PS concentration up to 1g/L would result in an increase in the NBB degradation rate. It was found that the PS-enhanced effect was strongly operating parameters dependent. The positive effect of PS decreased with increasing power and the best enhancing effect was obtained for the lowest acoustic power. Correspondingly, the PS-enhanced effect was more remarkable at low frequency (20kHz) than that observed at high frequency ultrasound (585kHz). Nitrogen saturating gas gave the best enhanced effect of PS than argon and air atmospheres. Theoretical (computer simulation of bubble collapse) and experimental measurements of the yields of free radical generation under the different experimental conditions have been made for interpreting the obtained effects of PS on the sonochemical degradation of the dye pollutant. The experimental findings were attributed to the fact that radical-radical recombination reactions occur at faster rate than the radical-organic reaction when the concentration of free radicals is too high (at higher sonochemical conditions).

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A new colorimetric method for monitoring of rolling circle amplification was developed. At first H5N1 target hybrids with padlock probe (PLP) and then PLP is circularized upon the action of T4 ligase enzyme. Subsequently, the circular probe is served as a template for hyperbranched rolling circle amplification (HRCA) by utilizing Bst DNA polymerase enzyme. By improving the reaction, pyrophosphate is produced via DNA polymerization and chelates the Mg(2+) in the buffer solution. This causes change in solution color in the presence of hydroxy naphthol blue (HNB) as a metal indicator. By using pH shock instead of heat shock and isothermal RCA reaction not only the procedure becomes easier, but also application of HNB for colorimetric detection of RCA reaction further simplifies the assay. The responses of the biosensor toward H5N1 were linear in the concentration range from 0.16 to 1.20 pM with a detection limit of 28 fM.

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In this work, the sonochemical degradation of naphthol blue black (NBB), an acidic diazo dye, in water was investigated. The effects of several operating parameters such as initial NBB concentration, acoustic intensity, ultrasonic frequency, nature of the dissolved gas and solution pH on the degradation of the dye were carried out. The obtained results showed that ultrasound completely destroyed NBB (5 mg L(-1)) after 45 min of sonication and most of the chemical oxygen demand was eliminated after 90 min of treatment. It was found that the initial rate of sonolytic degradation increased with increasing the initial NBB concentration. The fitting of the experimental data by a heterogeneous Langmuir-kinetics model showed that NBB degraded mainly at the interfacial region of the bubble by hydroxyl radical (OH) attack. The degradation rate of the dye increased substantially with increasing acoustic intensity in the range of 0.44-3.58 W cm(-2) and decreased with increasing frequency in the range of 585-1140 kHz. The rate of NBB degradation decreased in the order of Ar>air>N2. The significant degradation was achieved in acidic conditions (pH 2) where the initial degradation rate was 1.37 and 1.66 higher than those observed at pH 6 and pH 10, respectively.

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In the present work, comprehensive experimental and numerical investigations of the effects of frequency and acoustic intensity on the sonochemical degradation of naphthol blue black (NBB) in water have been carried out. The experiments have been examined at three frequencies (585, 860 and 1140 kHz) and over a wide range of acoustic intensities. The observed experimental results have been discussed using a more realistic approach that combines the single bubble sonochemistry and the number of active bubbles. The single bubble yield has been predicted using a model that combines the bubble dynamics with chemical kinetics consisting of series of chemical reactions (73 reversible reactions) occurring inside an air bubble during the strong collapse. The experimental results showed that the sonochemical degradation rate of NBB increased substantially with increasing acoustic intensity and decreased with increasing ultrasound frequency. The numerical simulations revealed that NBB degraded mainly through the reaction with hydroxyl radical (OH), which is the dominant oxidant detected in the bubble during collapse. The production rate of OH radical inside a single bubble followed the same trend as that of NBB degradation rate. It increased with increasing acoustic intensity and decreased with increasing frequency. The enhancing effect of acoustic intensity toward the degradation of NBB was attributed to the rise of both the individual chemical bubble yield and the number of active bubbles with increasing acoustic intensity. The reducing effect of frequency was attributed to the sharp decrease in the chemical bubble yield with increasing frequency, which would not compensated by the rise of the number of active bubbles with the increase in ultrasound frequency.