There has been a considerable development in microfluidic based immunodiagnostics over the past few years which has greatly favored the growth of novel point-of-care-testing (POCT). However, the realization of an inexpensive, low-power POCT needs cheap and disposable microfluidic devices that can perform autonomously with minimum user intervention. This work, for the first time, reports the development of a new microchannel capillary flow assay (MCFA) platform that can perform chemiluminescence based ELISA with lyophilized chemiluminescent reagents. This new MCFA platform exploits the ultra-high sensitivity of chemiluminescent detection while eliminating the shortcomings associated with liquid reagent handling, control of assay sequence and user intervention. The functionally designed microchannels along with adequate hydrophilicity produce a sequential flow of assay reagents and autonomously performs the ultra-high sensitive chemiluminescence based ELISA for the detection of malaria biomarker such as PfHRP2. The MCFA platform with no external flow control and simple chemiluminescence detection can easily communicate with smartphone via USB-OTG port using a custom-designed optical detector. The use of the smartphone for display, data transfer, storage and analysis, as well as the source of power allows the development of a smartphone based POCT analyzer for disease diagnostics. This paper reports a limit of detection (LOD) of 8 ng/mL by the smartphone analyzer which is sensitive enough to detect active malarial infection. The MCFA platform developed with the smartphone analyzer can be easily customized for different biomarkers, so a hand-held POCT for various infectious diseases can be envisaged with full networking capability at low cost.
Two feeding experiments were conducted to evaluate if methionine is limiting in practical grow-out diets for tilapia, Oreochromis spp. Four diets containing 32% protein and 5% lipid were designed to compare the use of diets high in dehulled solvent-extracted soybean meal (DSESM) and expeller pressed soybean meal (EPSM) compared with a diet containing 6% fish meal (FM). Tilapia (4.78 6 0.07 g, mean 6 SD) were randomly stocked into twelve 600-L flow-through tanks at 20 fish per tank. After 6 wk, there were no notable trends or statistically significant differences (P . 0.05) in final mean weight, survival rate, and feed conversion ratio (FCR) among the treatments. Because results of this study indicated that DSESM could totally replace FM in practical diets for juvenile tilapia, a second batch of diets were formulated using other protein sources. Typical levels of cottonseed meal (CSM), DSESM, and meat and bone meal (MBM) were used to evaluate whether methionine could be limiting. Two basal diet formulations were tested either without or with methionine supplement (0.06/100 g diet). The first diet contained 15% CSM, 27% DSESM, and 10% MBM and the second diet contained 15% CSM and 37% DSESM. These diets contained 28% protein and 5% lipid. Tilapia (3.90 6 0.05 g) were randomly stocked into twelve 60-L glass aquaria of a recirculation system at 18 fish per aquarium for 5 wk and then moved to the 600-L flow-through tanks for five more weeks. After 10 wk, there were no statistically significant differences (P . 0.05) in final mean weight, survival rate, and FCR among the four treatments. Results of the present study indicated that DSESM and EPSM could totally replace FM's inclusion rate in commercial diets for juvenile tilapia. Furthermore, methionine did not appear to be limiting in practical diets using typical levels of CSM, DSESM, and MBM as primary protein sources.
Two feeding experiments were conducted to confirm methionine requirement in practical diets of juvenile Nile tilapia, Oreochromis niloticus. Test diets used in both experiments contained 414 kcal gross energy, 28 g protein, and 5 g lipid per 100 g diet. In the first experiment, seven diets were made using cottonseed meal (CSM), dehulled solvent‐extracted soybean meal (DSESM), and gelatin as intact protein sources. Methionine was added to five of these diets at 0.03 or 0.06% increments to produce methionine levels ranging from 0.33 to 0.57% of the diet. Each diet was fed to four replicate groups of male juvenile Nile tilapia (5.62 ± 0.13 g) in a recirculation system for 8 wk. Broken‐line regression analysis of weight gain indicated that methionine requirement of juvenile Nile tilapia was 0.49% of the diet or 1.75% of dietary protein at cystine level of 0.45% of the diet. The second experiment was designed based on methionine requirement determined in the first experiment and also contained seven test diets. The first six diets contained CSM and DSESM as protein sources. Methionine was added to five of these diets at an increasing rate of 0.06% to produce methionine levels ranging from 0.49 to 0.79% of the diet. In the last diet (Diet 7), a portion of DSESM was replaced by gelatin to reduce methionine level to 0.33% of the diet in order to test whether methionine is limited. Each diet was also fed to four replicate groups of male juvenile Nile tilapia (2.32 ± 0.06 g) in a recirculation system for 9 wk. At the termination of the second experiment, there were no significant differences in terms of weight gain, survival, and feed efficiency ratio (FER) among the fish fed the first six diets. However, weight gain and FER of the fish fed these diets were significantly better than those fed Diet 7, confirming the methionine requirement value as has been determined in the first experiment.
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