A simple pyrimidine-based fluorescent probe (R)-4-(anthracen-9-yl)-6- (naphthalen-1-yl)-1,6-dihydropyrimidine-2-amine (ANDPA) was synthesized through the greener one pot reaction and characterized by IR, NMR, and ESI-Mass. Glucose stabilized silver nanoparticles (Glu-AgNPs) were also synthesized and characterized using UV, IR, XRD, SEM, and TEM. When ANDPA was tagged with Glu-AgNPs, the fluorescent intensity of ANDPA decreased drastically. When the monoclonal antibody (Ab) [immunoglobulin G (IgG)] of Pseudomonas aeruginosa (PA) was attached with ANDPA/Glu-AgNPs, the original intensity of the probe was recovered with minimal enhancement at 446 nm. On further attachment of PA with ANDPA/Glu-AgNPs/PA, the fluorescence intensity of the probe was enhanced obviously at 446 nm with red shift. This phenomenon was further supported by SEM and TEM. The linear range of detection is from 8 to 10 CFU/mL, and LOD is 1.5 CFU/mL. The immunosensor was successfully demonstrated to detect Pseudomonas aeruginosa in water, soil, and food products like milk, sugar cane, and orange juices.
Scale formation in heat exchanger tube reduces heat transfer efficiency and enhances corrosion. Scale formation in cooling water is due to many factors including pH, temperature, salt etc. In this study, microbiological aspects of scale formation and their role on corrosion are presented. The calcium precipitating bacteria (CPB) were isolated from the scales collected from heat exchanger tube in a gas turbine power station using B4 medium. The dominant CPB was isolated and identified using 16s rRNA sequencing, and the phylogenetic analysis reveals that the predominant bacteria were Serratia sp. (FJ973548), Enterobacter sp. (FJ973549, FJ973550), and Enterococcus sp. (FJ973551). The nature of crystal deposits of bacteria has been explained. The corrosion behavior of CPB on mild steel was studied by the electrochemical method (polarization and impedance), and the biogenic calcium scale formations in CPB were analyzed by XRD method. The scale formation by bacteria reduced the cathodic corrosion current, where resistance was lower in the presence of bacteria. It is claimed that the CPB is one of the causative factor for scale formation and corrosion in cooling water system.
Surveillance is a prime requisite for controlling arthropod vectors like mosquitoes that transmit diseases such as malaria, dengue and chikungunya. Carbon dioxide (CO2) is one of the main cues from vertebrate breath that attracts mosquitoes towards the host. Hence, CO2 is used as an attractant during surveillance of mosquitoes either from commercial cylinders or dry ice for mosquito traps. In the present study, the biogenic carbon dioxide production was optimized with different carbon sources such as glucose, simple sugar and jaggery with and without yeast peptone dextrose (YPD) media using commercial baker's yeast. The results showed that yeast produced more biogenic CO2 with simple sugar as compared to other carbon sources. Further substrate concentration was optimized for the continuous production of biogenic CO2 for a minimum of 12 h by using 10 g of baker's yeast with 50 g of simple sugar added to 1.5 l distilled water (without YPD media) in a 2-l plastic bottle. This setup was applied in field condition along with two different mosquito traps namely Mosquito Killing System (MKS) and Biogents Sentinel (BGS) trap. Biogenic CO2 from this setup has increased the trapping efficiency of MKS by 6.48-fold for Culex quinquefasciatus, 2.62-fold for Aedes albopictus and 1.5-fold for Anopheles stephensi. In the case of BGS, the efficiency was found to be increased by 3.54-fold for Ae. albopictus, 4.33-fold for An. stephensi and 1.3-fold for Armigeres subalbatus mosquitoes. On the whole, plastic bottle setup releasing biogenic CO2 from sugar and yeast has increased the efficiency of MKS traps by 6.38-fold and 2.74-fold for BGS traps as compared to traps without biogenic CO2. The present study reveals that, among different carbon sources used, simple sugar as a substance (which is economical and readily available across the world) yielded maximum biogenic CO2 with yeast. This setup can be used as an alternative to CO2 cylinder and dry ice in any adult mosquito traps to enhance their trapping efficiency of a mosquito surveillance programme.
Background & objectives:Botulism, a potentially fatal paralytic illness, is caused by the botulinum neurotoxins (BoNTs) secreted by Clostridium botulinum. It is an obligate anaerobic, Gram-positive, spore-forming bacterium. BoNTs are classified into seven serotypes based on the serological properties. Among these seven serotypes, A, B, E and, rarely, F are responsible for human botulism. The present study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA)-based detection system for the detection of BoNT/E.Methods:The synthetic gene coding the light chain of BoNT serotype E (BoNT/E LC) was constructed using the polymerase chain reaction primer overlapping method, cloned into pQE30UA vector and then transformed into Escherichia coli M15 host cells. Recombinant protein expression was optimized using different concentrations of isopropyl-β-D-1-thiogalactopyranoside (IPTG), different temperature and the rBoNT/E LC protein was purified in native conditions using affinity column chromatography. The purified recombinant protein was checked by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and further confirmed by western blot and matrix-assisted laser desorption ionization-tandem time-of-flight (MALDI-TOF). Polyclonal antibodies were generated against rBoNT/E LC using Freund's adjuvant in BALB/c mice and rabbit. Sandwich ELISA was optimized for the detection of rBoNT/E LC and native crude BoNT/E, and food matrix interference was tested. The developed antibodies were further evaluated for their specificity/cross-reactivity with BoNT serotypes and other bacterial toxins.Results:BoNT/E LC was successfully cloned, and the maximum expression was achieved in 16 h of post-induction using 0.5 mM IPTG concentration at 25°C. Polyclonal antibodies were generated in BALB/c mice and rabbit and the antibody titre was raised up to 128,000 after the 2nd booster dose. The developed polyclonal antibodies were highly specific and sensitive with a detection limit about 50 ng/ml for rBoNT/E LC and 2.5×103 MLD50 of native crude BoNT/E at a dilution of 1:3000 of mouse (capturing) and rabbit (revealing) antibodies. Further, different liquid, semisolid and solid food matrices were tested, and rBoNT/E LC was detected in almost all food samples, but different levels of interference were detected in different food matrices.Interpretation & conclusions:There is no immune detection system available commercially in India to detect botulism. The developed system might be useful for the detection of botulinum toxin in food and clinical samples. Further work is in progress.
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