Autofluorescence changes of tomato surface tissues during overripening.

We investigated the autofluorescence of tomato surface tissues during overripening at 25 °C for 13 days. Microscopic images and fluorescence spectra of tissues, including the epidermis and cuticle, were examined (excitation at 360 nm), revealing that the autofluorescence changes were related to the epidermis, particularly the fluorophores in the cuticle.

Chick Embryo Growth Modeling Using Near-Infrared Sensor and Non-Linear Least Square Fitting of Egg Opacity Values.

Non-destructive monitoring of chick embryonic growth can provide vital management insights for poultry farmers and other stakeholders. Although non-destructive studies on fertility, hatching time and gender have been conducted recently, there has been no available method for embryonic growth observation, especially during the second half of incubation. Therefore, this work investigated the feasibility of using near-infrared (NIR) sensor-based egg opacity values-the amount of light lost when passing through the egg-for indirectly observing embryo growth during incubation. ROSS 308 eggs were selected based on size, mass and shell color for this experiment. To estimate the embryo size precisely, we fit various mathematical growth functions during incubation, based on the opacity value of incubated eggs. Although all the growth models tested performed similarly in fitting the data, the exponential and power functions had better performances in terms of co-efficient of determination (0.991 and 0.994 respectively) and RMSE to explain embryo growth during incubation. From these results, we conclude that the modeling paradigm adopted provides a simple tool to non-invasively investigate embryo growth. These models could be applied to resolving developmental biology, embryonic pathology, industrial and animal welfare issues in the near future.

A Noise Tolerant Spread Spectrum Sound-Based Local Positioning System for Operating a Quadcopter in a Greenhouse.

Quadcopters are beginning to play an important role in precision agriculture. In order to localize and operate the quadcopter automatically in complex agricultural settings, such as a greenhouse, a robust positioning system is needed. In previous research, we developed a spread spectrum sound-based local positioning system (SSSLPS) with a 20 mm accuracy within a 30 × 30 m greenhouse area. In this research, a noise tolerant SSSLPS was developed and evaluated. First, the acoustic noise spectrum emitted by the quadcopter was documented, and then the noise tolerance properties of SSSounds were examined and tested. This was done in a greenhouse with a fixed quadcopter (9.75 N thrust) with the positioning system mounted on it. The recorded quadcopter noise had a broadband noise compared to the SSSound. Taking these SSSound properties into account, the noise tolerance of the SSSLPS was improved, achieving a positioning accuracy of 23.2 mm and 31.6 mm accuracy within 12 × 6 m for both Time-division Multiple Access (TDMA) and Frequency-division Multiple Access (FDMA) modulation. The results demonstrate that the SSSLPS is an accurate, robust positioning system that is noise tolerant and can used for quadcopter operation even within a small greenhouse.

Broadband dielectric spectroscopy of glucose aqueous solution: Analysis of the hydration state and the hydrogen bond network.

Recent studies of saccharides' peculiar anti-freezing and anti-dehydration properties point to a close association with their strong hydration capability and destructuring effect on the hydrogen bond (HB) network of bulk water. The underlying mechanisms are, however, not well understood. In this respect, examination of the complex dielectric constants of saccharide aqueous solutions, especially over a broadband frequency region, should provide interesting insights into these properties, since the dielectric responses reflect corresponding dynamics over the time scales measured. In order to do this, the complex dielectric constants of glucose solutions between 0.5 GHz and 12 THz (from the microwave to the far-infrared region) were measured. We then performed analysis procedures on this broadband spectrum by decomposing it into four Debye and two Lorentz functions, with particular attention being paid to the ß relaxation (glucose tumbling), δ relaxation (rotational polarization of the hydrated water), slow relaxation (reorientation of the HB network water), fast relaxation (rotation of the non-HB water), and intermolecular stretching vibration (hindered translation of water). On the basis of this analysis, we revealed that the hydrated water surrounding the glucose molecules exhibits a mono-modal relaxational dispersion with 2-3 times slower relaxation times than unperturbed bulk water and with a hydration number of around 20. Furthermore, other species of water with distorted tetrahedral HB water structures, as well as increases in the relative proportion of non-HB water molecules which have a faster relaxation time and are not a part of the surrounding bulk water HB network, was found in the vicinity of the glucose molecules. These clearly point to the HB destructuring effect of saccharide solutes in aqueous solution. The results, as a whole, provide a detailed picture of glucose-water and water-water interactions in the vicinity of the glucose molecules at various time scales from sub-picosecond to hundreds of picoseconds.

Assessment of chalkiness index of Sake rice using transmission imaging.

The higher chalkiness level of the white core kernel is prone to breakage during the high degree polishing. So, grading white core kernel based on chalkiness level is crucial to making premium quality Sake (rice wine) in the brewing industry. The chalkiness level in the white core kernel is currently performed destructively. Thus, a chalkiness index is required to assess the level in the white core kernel. This research assesses the white core rice kernel based on the chalkiness index non-destructively. Here, the optical transmission property in the visible to near-infrared (VIS-NIR) region of rice was measured using a V-670 spectrophotometer equipped with an integrating sphere to investigate the variation of chalkiness level rice samples. The images were then acquired by transmission mode of four types of intact Sake rice kernel using blue light-emitting diodes (LEDs), green, red, and NIR LEDs in which the peak wavelength of the LEDs was 465 nm, 525 nm, 630 nm, and 830 nm, respectively. The result indicates that the rice samples were more penetrated and better visualized chalkiness by light in the NIR region. Therefore, the wavelength region in NIR showed better discrimination between transparent and opaque parts in white core's Sake rice. Furthermore, the proposed chalkiness index was inversely correlated with the gray-level intensity of the transmittance image. This gray value was significantly correlated (R2 = 0.89) with the chalkiness index in the NIR region. So, gray values of NIR transmittance images were identified as sensitive for chalkiness index, which would be applied for sorting the white core kernel with different levels of chalkiness in the Sake brewing industry.

Japanese dace (Tribolodon hakonensis) fish freshness estimation using front-face fluorescence spectroscopy coupled with chemometric analysis.

Although fish and its related products are good sources of protein and unsaturated fatty acids, like omega-3 in the human diet, their shelf-life is limited by biochemical and microbial changes. In this study, a front-face fluorescence spectroscopy technique was used to acquire Excitation-emission matrices (EEM) to monitor Japanese dace (Tribolodon hakonensis) fish freshness degradation during storage. EEM of Japanese dace fish parts (intact eyeball and surface-containing scales), excitation from 220 to 585 nm and emissions from 250 to 600 nm, were measured at different times during storage. To simplify the acquired complex spectra datasets from each fish part, the variables were reduced to those that were only significant/important (those with higher positive or negative correlation) for K value prediction, and as an index of freshness. Partial least square regression (PLSR) results demonstrated that combining the fluorescence EEM of the eyeball and surface-containing scales the best monitoring of fish freshness; excitation at 280 and 350 nm for both the eyeball and surface-containing scales, with 2.84 and 0.96 as RMSE and R2, respectively. These findings demonstrate that multiple excitation fluorescence approaches can be convenient for the freshness evaluation of fish.

UV excited fluorescence image-based non-destructive method for early detection of strawberry (Fragaria × ananassa) spoilage.

The soon spoiled strawberries need to be classified from healthy fruits in an early stage. In this research, a machine vision system is proposed for inspecting the quality of strawberries using ultraviolet (UV) light based on the excitation-emission matrix (EEM) results. Among the 100 fruits which were harvested and stored under 10 °C condition for 7 days, 7 fruits were confirmed to be spoiled by using a firmness meter. The EEM results show the fluorescence compound contributes to a whitish surface on the spoiled fruits. Based on the EEM results, UV fluorescence images from the bottom view of strawberries were used to classify the spoiled fruits and healthy fruits within 1 day after harvest. These results demonstrate the UV fluorescence imaging can be a fast, non-destructive, and low-cost method for inspecting the soon spoiled fruits. The proposed index related to the spoiling time can be a new indicator for qualifying strawberry.

Research Note: Nondestructive detection of super grade chick embryos or hatchlings using near-infrared spectroscopy.

Some unresolved questions in poultry science were addressed: what determines the yield of chick embryos or hatchlings; what kind of influence does egg yolk content have on embryonic development; and how to detect eggs producing super grade chicks? Since the yolk acts as a vital energy and nutrient reservoir for embryos, we hypothesized that a higher yolk content of similar sizes eggs would play an important role in embryo or chick viability during incubation, as well as at hatch. As experimental sample, we used ROSS 308 (broiler line) and a nondestructive spectroscopic absorbance method. The influence of high yolk content to embryonic heartbeat and chick yield (i.e., chick weight/egg weight) were then investigated. Embryonic heartbeat signal was measured indirectly using a prototype near-infrared sensor during incubation period. A positive influence was found in both cases. Similar size eggs with higher yolk content were found to significantly (P-value < 0.05) promote higher chick yield at hatch. This methodology may have the potential to be used to precision poultry production system, ornithology, developmental, or evolutionary biology in the near future.

Prediction of protein and oil contents in soybeans using fluorescence excitation emission matrix.

To investigate the potential of fluorescence spectroscopy in evaluating soybean protein and oil content, excitation emission matrix (EEM) was measured on 34 samples of soybean flours using a front-face measurement, and the accuracy of the protein and oil content prediction was evaluated. The EEM showed four main peaks at excitation/emission (Ex/Em) wavelengths of 230/335, 285/335, 365/475, and 435/495 nm. Furthermore, second derivative synchronous fluorescence (SDSF) spectra were extracted from the EEMs, and partial least square regression and support vector machine models were developed on each of the EEMs and SDSF spectra. The R2 values reached 0.86 and 0.74 for protein and oil, respectively. From the loading spectra, fluorescence at Ex/Em of 230-285/335 nm and 350/500 nm mainly contribute to the protein and oil content prediction, respectively. Those results revealed the potential of fluorescence spectroscopy as a tool for a rapid prediction of soybean protein and oil content.

Thermal oxidation assessment of Italian extra virgin olive oil using an UltraViolet (UV) induced fluorescence imaging system.

Extra virgin olive oil is a high-quality product with profound health benefits but is susceptible to degradation due to oxidation. Environmental conditions such as temperature, oxygen, and light, promote the oxidation process. From this perspective thermal oxidation stability is of primary concern in terms of food quality and safety. The ability to resist oxidation ensures continued nutritional and economic value. In this study, the thermal oxidation stability of four mono-cultivars of extra virgin olive oil from four different regions of Italy was studied. The samples underwent thermal treatment at 120 °C with measurements taken at regular time intervals over 180 min. To develop a simplified imaging system, the fluorescence characteristics of the samples during thermal exposure were measured using front-face fluorescence and transmittance spectroscopy in order to assess the changes that occur due to thermal exposure. Standard quality indices including; Peroxide value, acidity, K232, and K270, were also measured following IOC (International Olive Council) procedures. Image processing of both color and fluorescence images was done to ascertain cultivar responses to the thermal treatment. Fluorescence peaks associated with polyphenols, oxidation products, and chlorophyll were identified and monitored, and a comparison made between the different cultivars. Fluorescence peaks were observed at emission wavelengths 435, 465, and 570 nm, which are suspected to be products of oxidation and hydrolysis, respectively. The cultivars with a higher concentration of polyphenols showed greater resistance to the formation of oxidation products; an indication that they have a higher thermal stability. The B channel of the RGB color space was identified as being sensitive to changes in UltraViolet (UV) induced fluorescence images due to thermal exposure, and to enable the monitoring of the thermal stability of the different cultivars of extra virgin olive oil.

A Nondestructive Eggshell Thickness Measurement Technique Using Terahertz Waves.

Eggshells play a number of important roles in the avian and reptile kingdom: protection of internal contents and as a major source of minerals for developing embryos. However, when researching these respective roles, eggshell thickness measurement remains a bottleneck due to the lack of a non-destructive measurement techniques. As a result, many avian and reptile research protocols omit consideration of eggshell thickness bias on egg or embryo growth and development. Here, we validate a non-destructive method to estimate eggshell thickness based on terahertz (THz) reflectance spectroscopy using chicken white coloured eggs. Since terahertz waves are reflected from outer air-eggshell interface, as well as the inner eggshell-membrane boundary, the resulting interference signals depend on eggshell thickness. Thus, it is possible to estimate shell thickness from the oscillation distance in frequency-domain. A linear regression-based prediction model for non-destructive eggshell thickness measurement was developed, which had a coefficient of determination (R2) of 0.93, RMSEP of 0.009, RPD of 3.45 and RER 13.67. This model can estimate eggshell thickness to a resolution of less than 10 µm. This method has the potential to expand the protocols in the field of avian and reptile research, as well as be applied to industrial grading of eggs.

Quantification of starch content in germinating mung bean seedlings by terahertz spectroscopy.

To investigate the potential of terahertz spectroscopy to monitor and quantify starch in plants, terahertz spectra (3.0-13.5 THz) of mung bean plants 1-7 days after germination were examined and compared to those of starch and its constituent saccharides (standard reagents). Day 1 seedlings showed similar spectral features with standard starch, and absorption peaks gradually disappeared in the subsequent 6 day growth period. To interpret this result and identify useful peaks for starch quantification, standard starch and day 1 seedlings were hydrolyzed by α-amylase in vitro. Since both standard starch and seedlings showed that absorption peak at 9.0 THz disappeared after amylase hydrolysis, this peak is sensitive to changes in starch. Additionally, intensity of this peak was correlated with starch content as quantified by chemical analysis (r = 0.98). Our results indicate terahertz spectra of seedlings can provide an identifiable peak that is attributed to starch and not affected by the constituent saccharides.

Hydration and Hydrogen Bond Network of Water during the Coil-to-Globule Transition in Poly(N-isopropylacrylamide) Aqueous Solution at Cloud Point Temperature.

Aqueous solutions of poly(N-isopropylacrylamide), P-NIPAAm, exhibit a noticeable temperature responsive change in molecular conformation at a cloud point temperature (Tcp). As the temperature rises above Tcp, the extended coil-like P-NIPAAm structure changes into a swollen globule-like conformation as hydration levels decrease and hydrophobic interactions increase. Though water plays an important role in this coil-to-globule transition of P-NIPAAm, the behavior of water molecules and the associated hydrogen-bond (HB) network of the surrounding bulk water are still veiled in uncertainty. In this study, we elucidate changes in the hydration state and the dynamical structure of the water HB network of P-NIPAAm aqueous solutions during the coil-to-globule transition by analyzing the complex dielectric constant in the terahertz region (0.25-12 THz), where bulk water reorientations and intermolecular vibrations of water can be selectively probed. The structural properties of the water HB network were examined in terms of the population of the non-HB water molecules (not directly engaged in the HB network or hydrated to P-NIPAAm) and the tetrahedral coordination of the water molecules engaged in the HB network. We found the hydration number below Tcp (≈10) was decreased to approximately 6.5 as temperature increased, in line with previous studies. The HB network of bulk water becomes more structured as the coil-to-globule phase transition takes place, via decreases in non-HB water and reduction in the orderliness of the tetrahedral HB architecture. Together these results indicate that the coil-to-globule transition is associated with a shift to hydrophobic-dominated interactions that drive thermoresponsive structural changes in the surrounding water molecules.