Detection of internal wheat seed infestation by Rhyzopertha dominica using X-ray imaging

Standardization for grain grades has been established in most countries to maintain the quality of a crop until it reaches consumers. Different methods have been investigated for their potential to detect insect infestations in grain destined for domestic and export markets. The potential of detecting infestations caused by Rhyzopertha dominica in wheat kernels using a real-time soft X-ray method was determined in this study. Artificially infested wheat kernels were incubated at 30°C and 70% relative humidity and X-rayed sequentially for larval, pupal, and adult stages of R. dominica. Algorithms were used to extract histogram features, textural features, and histogram and shape moments from the X-ray images of wheat kernels. A backpropagation neural network (BPNN) and statistical classifiers were used to identify uninfested and infested kernels using the 57 extracted features. The BPNN correctly identified all uninfested and infested kernels and more than 99% of kernels infested by R. dominica larvae. The classification accuracies determined by the BPNN were higher using all 57 features than when using the histogram and textural features separately. The BPNN performed better than the parametric and non-parametric classifiers in the identification of uninfested and infested kernels by different stages of R. dominica.     

Effects of insect-infested kernels on bulk flow properties of wheat

Insect damage to kernels during storage affects the processing quality of wheat and as well as bulk wheat properties, which in turn causes hopper flow problems such as funnel flow, ratholing, arching, or flushing. This study focused on kernel damage by Rhyzopertha Dominica F. (lesser grain borer), one of the most commonly found insects in wheat, and resulting changes in physical properties, such as bulk density, tapped density, true density, and angle of repose, and in flow properties, such as basic flowability energy, stability, aeration, compressibility, and permeability. Bulk and tapped densities of sound hard red winter wheat kernels and infested wheat kernels were about 720 kg/m³ and 775 kg/m³, respectively. Compressibility index (CI), Hausner ratio (HR), and angle of repose of infested wheat kernels were higher than for sound hard red winter wheat kernels, indicating decreased flowability. Grain dust and insect-infested kernels could form localized compacted areas within the grain bins, resulting in uneven flow during discharge. Results from this study indicate that the presence of insect-infested kernels could lead to arching and bridge formation in grain bins, thus affecting the flow characteristics of bulk wheat.     

A comparison of screening methods for insect contamination in wheat

In collaboration with the United States Department of Agriculture and a number of major milling companies, the “Insect-Detect” immunoassay for analyzing insect contamination in grains has been compared with three more traditional methods, X-ray analysis, cracking and flotation, and the insect fragment test (IFT). Testing was carried out in blind fashion using clean wheat samples that were spiked with differing numbers of grain kernels infested with late instar larvae of the granary weevil (Sitophilus granarius (L.)). Three different laboratories analyzed the samples for each of the four methods. The collaborative trials showed that the insect immunoassay clearly provided the most accurate measurement of actual insect infestation, followed by X-ray analysis. While both cracking and flotation and IFT procedures provided a general measure of contamination, they showed much greater variability.     

Detection of internal insects in wheat using a conductive roller mill and estimation of insect fragments in the resulting flour

A laboratory roller mill that monitors the conductance of kernels that pass through it was tested for its ability to estimate the number of insect fragments in flour after milling. This system can test a kilogram of whole wheat in approximately 1 min and requires little sample preparation. Hard red winter wheat samples were infested with lesser grain borers and stored at 24 °C. Infestations ranged from 12 to over 2000 infested kernels per 1 kg or per 30,000 kernels. After crushing of samples in the conductance instrument, the samples were milled into flour and sub-samples were sent to two laboratories for insect fragment analysis. The insect fragments were proportional to the number of detection incidences obtained using the conductance instrument and X-ray images. Insect fragment counts per 50 g of flour ranged from 0 to over 5000. For insect fragment counts from 0 to 250, correlations between fragment counts and conductance mill detection were 0.75 and 0.80 from two separate cereal chemistry laboratories. Therefore, the conductance mill is potentially a good method for testing incoming grain for live internally infesting insects; it is able to test 1 kg of grain in about 1 min and can detect low levels (as low as three insects) of live internal infestations in a 1- or 2-kg sample.     

Hermetic storage of pearl millet, Pennisetum americanum (L.) Leeke; laboratory studies

The effect of airtight storage of pearl millet, Pennisetum americanum (L.) Leeke, on intergranular atmosphere, insects, mold infection, grain viability and moisture content (m.c.) was investigated in the laboratory. Six hermetically sealed 19-1 cans were used to store millet of three moisture contents (9.9, 12.8 and 15.6%) for 76 days at 27 ± 1°C. One can of grain of each m.c. was infested with 150 adult Sitophilus oryzae L./kg; the others remained uninfested. Carbon dioxide concentrations increased and oxygen concentrations decreased in all cans infested with insects, and in uninfested grain of 15.6% m.c. The changes in atmospheric concentrations were slower in the uninfested grain than in the infested 15.6% m.c. grain. No live insects were found in any treatment, percentages of mold-infested kernels were not significantly different among treatments, and m.c. did not change due to treatment or time. Kernel viability decreased significantly only in the 15.6% m.c. gram. All grain appeared normal; but the 15.6% m.c. grain had a slightly sour odor.     

Wheat-bug damage in New Zealand wheats: The feeding mechanism of nysius huttoni and its effect on the morphological and physiological development of wheat

Adult Nysius huttoni White were placed in separate cages containing cultivars Rongotea and Karamu at the late anthesis, watery ripe and milky ripe stages of development. Between 84 and 99 % of the matured kernels were injured with the characteristic visible markings of bug-damaged wheat. All the samples contained strong wheat-bug proteinase activity as shown by the incubated SDS-sedimentation test and the disappearance of HMW glutenin subunits from electrophoretograms. Grain infested at late anthesis was most severely affected with shrivelled kernels, high screenings, protein, free amino acids and α-amylase, and low kernel weight, germination capacity and carbohydrate content. Grain infested at the watery ripe and milky ripe stages had values for the above properties closer to uninfested control wheat values. It was suggested that N huttoni sucked sap from lateral sieve tubes in the wheat ovary at late anthesis causing severe disruption to physiological development of the grain. Two commercial lines of wheat showed some characteristics of this effect.     

基于软X射线与低场核磁检测小麦隐蔽性害虫玉米象

针对小麦内部隐蔽性害虫玉米象(Sitophilus zeamais)难以检出的问题,本文将高清软X射线与低场核磁两种检测技术相结合,通过高清软X射线拍摄的图片,观察玉米象在小麦内部的整个生长周期,提取图片纹理特征,用线性判别分析(linear discriminant analysis,LDA)与二次判别分析(quadratic discriminant analysis,QDA)算法进行分类判别,并对被不同虫态玉米象感染的小麦进行低场核磁检测。研究结果表明:在12%水分含量小麦中,玉米象的生长周期大约为36 d,LDA与QDA模型对未感染小麦与不同感染阶段小麦进行单独分类判别时,准确率都达到了95%以上,对卵期以及幼虫具有较高的分类准确率。根据小麦被玉米象感染后特征峰值比例P2b与P22的变化,可以用来作为小麦是否受玉米象感染定性判断的依据。     

Comparison of seven methods for stored cereal losses to insects for their application in rural conditions

There are several methods for assessing harvest losses during storage with varying degrees of accuracy, reliability and practical application, however, field conditions may limit their use among subsistence farmers and traders in developing nations. We compared seven standard grain loss methods during natural and controlled infestation by Sitophilus oryzae L. in wheat and barley to select the best procedures for use in farm storage. Methods tested were: 1) visual inspection of infested lots, 2) uncorrected weight loss, 3) modified standard volume/weight ratio, 4) grain count and weight, 5) percentage of damaged grains converted to weight loss, 6) one thousand grain mass, and 7) one thousand grain mass including dust. Previously disinfested cereal lots (wheat, barley, 500 g fresh w) were exposed to 20 newly emerged adult S. oryzae for 90 d at 28 °C and 70% r.h. Naturally- infested wheat lots were also monitored under the same conditions. Before and after this period, fresh and dry weight, number, moisture content of uninfested and damaged grains, weight of 1 L of seeds and dust produced by insect activity, fresh and dry weight of 1000 kernels, and number of adult weevils was determined and applied to appropriate equations of methods (1), (2), (3), (4), (5), (6) and (7). Grain mass loss estimations varied by wide margins (9.3 ± 1.3% to 25.8 ± 2.3% in barley, 2.2 ± 0.7% to 12.5 ± 2.5% in wheat) depending on method employed, suggesting the need for careful selection of the most appropriate procedure under field conditions and farmer/trader interests. While accurate procedures such as method 7 provided dependable results, methods 3 and 5 appear most practical for wheat and barley.     

The influence of Rhyzopertha dominica (F.) on the technological quality of cereal grains treated with diatomaceous earth

Laboratory bioassays (temperature 26 ± 1 °C and 60 ± 5% r. h.) were performed to evaluate the insecticidal effect of three different diatomaceous earths (DE) against Rhyzopertha dominica (F.) in wheat, triticale and rye. In order to achieve this, the amount of damaged kernels, amount of debris, wet gluten content, gluten index and rheological properties were assessed in infested samples, infested samples treated with DE Protect-It and two DEs originating from Serbia (S-1 and S-2) and were compared to control (uninfested) samples using Chopin Mixolab. The most susceptible to beetle infestation were rye sample and wheat variety Planeta. The lowest weight of damaged grain was found after applying DE Protect-It to all grain types. The amount of damaged kernels, the amount of debris and loss of mass were the highestin sample treated with DE S-2. While infestation resulted in increase in the amount of damaged kernels and debris and decrease in wet gluten content, and provoked weakening of gluten network structure as well as starch damage, an addition of DE, especially Protect-It, resulted in grain samples of similar technological quality to control (uninfested) sample. The susceptibility of different cereals used in breadmaking to infestation by R. dominica, as well as their response to the insecticidal effect of diatomaceous earth largely varied. Both cereal type and quality, as measured by wet gluten content and gluten index, influenced grain response to infestation. Insecticidal effect of diatomaceous earth, especially DE S-1and DE S-2, was the least effective in terms of triticale. These findings suggest that the application of DE in cereals storage management might largely be affected with grain type as well as grain composition and technological quality.     

Detection of Rhyzopertha dominica larvae in stored wheat using ELISA: The impact of myosin degradation following fumigation

Hard red winter wheat kernels were infested with eggs of Rhyzopertha dominica. After 20 d, when the larvae reached the fourth instar, they were killed by exposing the infested kernels to phosphine gas for 24 h. The infested kernels were then divided into four portions and treated as follows: one portion was immediately frozen at −80 °C to avoid myosin degradation; the other three portions were kept at 32 °C and 65% relative humidity, and then frozen at −80 °C after 14, 28, and 56 d post-fumigation, respectively. Each treatment was replicated five times. Myosin was measured using a commercial enzyme linked immunosorbent assay (ELISA) method that specifically detects this protein (Biotect^®, Austin, TX). Myosin degradation was most rapid in the first 2 weeks after the larvae were killed, decreasing from 1.672 to 0.695 ng/well during this period (a 58.4% reduction). There were no significant differences in myosin degradation between samples that were 14, 28, and 56 d post-fumigation. Grain is often fumigated to control insects. Frequently, this occurs many weeks before the grain is milled and may be repeated during the storage period. Therefore, estimates using the ELISA test may underestimate internal insect infestation because of myosin degradation. Insect fragment estimates for previously fumigated grain could be underestimated by as much as 58%.     

Susceptibility of Ethiopian wheat varieties to granary weevil and rice weevil infestation at optimal and sub-optimal temperatures

Integrating varietal resistance with temperature manipulation during storage may provide a better option for protection of stored grains and may decrease reliance on the use of synthetic chemicals. The current study was conducted to determine the susceptibility of different varieties of wheat seed to the infestation by the granary weevil, Sitophilus granarius (L.), and rice weevil, Sitophilus oryzae (L.), at optimal (30 °C) and sub-optimal (19 °C) temperatures. Kernels of six wheat varieties namely, Danda'a, Digalu, ET-13-A2, Kakaba, Millennium, and Pavon-76 were examined over a period of 90 d. Significant interactions were detected between wheat varieties and storage temperature for progeny emergence, percentage of insect damaged kernels, grain weight loss, and amount of powder produced per gram of wheat. Kernels of Danda'a, infested with S. oryzae at 30 °C exhibited significantly lower mean progeny counts (13.3 live insects), lower percentage of grain weight loss (4.2%) and insect-damaged kernels (6.4%), and powder production (1.5 mg/g). Kernel weight and hardness index were negatively associated with percentage of insect damaged kernels and grain weight loss. Kernel diameter was positively associated with both of percentage of insect damaged kernels and grain weight loss. Wheat varieties with high Zeleny sedimentation values had lower percentage of insect-damaged kernels and grain weight loss. These results indicated that kernel weight, hardness index, and protein content are predominant factors contributing to wheat resistance against S. granarius and S. oryzae. The varieties Millennium and Danda'a can be considered with other integrated pest management approaches to reduce stored grain losses of wheat in Ethiopia.     

Detection of internally infested popcorn using electrically conductive roller mills

To detect popcorn kernels infested by the internal feeding stored-product insect pest Sitophilus zeamais, maize weevil, a laboratory roller mill was modified so that the electrical conductivity of the grain is measured while the kernels are milled between the rolls. When a kernel with a S. zeamais larvae inside is milled, the moisture from the crushed insect abruptly changes the conductivity of the test circuit. The objective of this study was to determine the potential of the modified conductance mill to detect popcorn infested with different developmental stages. Two laboratory milling units were tested that had differing sharpness, which affected the feed-rate through the rolls. One mill averaged 135 s to feed 1 kg of popcorn while the second mill with sharper teeth averaged 100 s to feed 1 kg of popcorn. Four popcorn varieties were evaluated, with their average kernel weight ranging from 12.5 g to 18.5 g per 100 kernels. Known numbers of infested popcorn kernels were added to 1 kg samples of popcorn. The slower feeding mill detected 81% of the pupae, 91% of the medium larvae, and 47% of the small larvae. The faster feeding mill detected 75% of the pupae, 80% of the medium larvae, and 43% of the small larvae. Our results indicate that the conductance mill is a good method for quickly evaluating popcorn samples for kernels infested with late stage larvae and pupae.     

Effect of the degree of fusariosis on the physical characteristics of individual wheat kernels

This work aims to determine dependencies of single‐kernel (SK) physical traits on the kernel degree of fusariosis. The measurements were taken on samples of grain of six wheat varieties cultivated during 1–3 years. For SK mass, volume and width this dependence resulted in lowered (by at least ten to more than 25% for mass and volume, and ca. 5 to more than 15% for width) values in Fusarium culmorum infested grain as compared with the non‐inoculated control. In addition, moderately infested kernels tended to be of higher mass and size than kernels with no symptoms of infestation. An unexpected effect was a lack of dependence of SK density on the degree of infestation. Moisture content was lower for infested kernels by 6–8%, hardness tended to increase a little for moderate infestation, while it noticeable decreased for the heaviest infestation.     

Thermal imaging to detect infestation by Cryptolestes ferrugineus inside wheat kernels

Canada's zero tolerance for live insects in grain received from farmers, and shipped to domestic and export buyers, has necessitated the development of an accurate insect detection method. An infrared thermal imaging system was developed to detect infestation by six developmental stages (four larval instars, pupae and adults) of Cryptolestes ferrugineus under the seed coat on the germ of the wheat kernels. The artificially infested wheat kernels were removed from the incubation room (30 °C), refrigerated (5 °C) for 60 s, maintained at ambient conditions for 20 s, and imaged using a thermal camera to identify each developmental stage (n=283). The means of the highest 5% and 10% of all temperature values on the surface of the grain were significantly higher (=0.05) for grains having young larvae inside and lower for grains having pupae inside. Temperature distribution on the surface of the infested kernels with different stages of C. ferrugineus was highly correlated with the respiration rate of each developmental stage (r=0.83–0.91). The overall classification accuracy for a quadratic function was 83.5% and 77.7% for infested and sound kernels, respectively, and for a linear function, it was 77.6% and 83.0% for infested and sound kernels, respectively, in pairwise discriminations. Thermal imaging has the potential to identify whether the grain is infested or not, but is less effective in identifying which developmental stage is present.     

Comparison of insect detection efficiency by different detection methods

Detection percentage of stored grain insects using different separation methods was determined. The stored grain insects tested were: Oryzaephilus surinamensis (L.), Rhyzopertha dominica (F.), Sitophilus oryzae (L.), and Tribolium castaneum (Herbst). To determine the detection percentage, 1 or 100 adults of the four insect species, and 1 or 100 young or old larvae of T. castaneum were introduced into wheat with 14 ± 1%, 16 ± 1%, or 18 ± 1% moisture content. The following four methods were used to separate the introduced insects from the infested wheat: Berlese funnel (Berlese), shaking grain on a metal sieve (Metal sieve), shaking grain on an acrylic device (Acrylic sieve), and microwave heating. For all detection methods, wheat moisture content did not influence the detection percentage. Detection method and insect species influenced the recovery percentage. For example, Metal sieve and Acrylic sieve methods had a higher recovery percentage of adults of O. surinamensis and T. castaneum than for S. oryzae and R. dominica. There was no significant difference in recovery percentage of the larvae and adults between the Metal sieve and Acrylic sieve methods, with the exception of weevils. The Berlese method had the lowest recovery percentage of introduced insects, with the exception of weevils, while the Microwave method had the highest or the second highest recovery percentage. Generally, the recovery percentage of the young larvae was lower than that of adults. The Microwave method could improve detection accuracy for larvae.     

Influence of the degree of fusariosis on technological traits of wheat grain

The paper aims to enlarge the current knowledge about the dependence of technological traits of wheat grain on its degree of infestation (DI) with fusariosis. Samples of grain of four wheat cultivars were obtained from naturally grown crops (control) and from crops inoculated with Fusarium culmorum from which three degrees of kernel infestation were selected. Samples were determined for thousand kernels weight (TKW), total protein, starch and wet gluten (WG) contents, Hagberg falling number (HFN) and sedimentation value (SV). Content of protein and wet gluten showed higher values for moderately infested kernels (up to 15 and 25%, respectively, when compared to control grain) than for kernels lightly or heavily infested. All the remaining traits had values lower than that for control grain and showed a decrease with increasing DI. Decrease for the heaviest infestation was on average: 8, 29, 31 and 34% for starch, TKW, HFN and SV, respectively.     

End-use quality characteristics of hermetically stored paddy

Four paddy varieties (Bg 352, Bg 300, Bg 358 and Bg 360) were stored in hermetic IRRI bags and common woven polyethylene bags (polybags) at room temperature either uninfested or infested with rice weevils (Sitophilus oryzae (L.)). After 9 months of storage, samples were tested for insect mortality, gas contents, moisture content (m.c.), thousand grain mass (TGM), porosity, hardness, whiteness, total milled rice yield (TMR), head rice yield (HRY), gelatinization temperature, amylose (AC), crude protein (CP), crude fat, free fatty acid (FFA), thiamine and niacin contents and sensory characteristics. These properties after storage were compared with their initial condition. The oxygen content dropped from 21% to 7% and 13.8% for infested and uninfested IRRI bag samples, respectively. The results showed that m.c., of the IRRI bag samples increased significantly (P < 0.05) by 5% when compared to the initial sample but it increased by 15% in polybag stored samples. After 9 months, dry matter loss (DM) was 65% higher in polybag than IRRI bag samples. Highest DM loss was observed in Bg 300 and the lowest DM loss was observed in Bg 358 and Bg 360. Paddy samples stored in IRRI bags showed reduced whiteness compared to polybag stored samples. Storage in IRRI bags significantly increased (P < 0.05) TMR, HRY, AC and sensory values compared to polybag samples. However, paddy samples stored in polybags significantly increased (P < 0.05) their CP and FFA contents while decreasing sensory values, thiamine and niacin contents. The FFA value of polybag samples was 2.5 times higher than IRRI bag samples. Hermetic storage of dry paddy improved overall paddy quality but different end-use quality parameters were observed in the two paddy grain types of short round (Bg 352 and Bg 300) or intermediate bold (Bg 358 and Bg 360).     

Detection of insect-damaged wheat kernels using near-infrared hyperspectral imaging

Insect damage in wheat adversely affects its quality and is considered one of the most important degrading factors in Canada. The potential of near-infrared (NIR) hyperspectral imaging for the detection of insect-damaged wheat kernels was investigated. Healthy wheat kernels and wheat kernels visibly damaged by Sitophilus oryzae, Rhyzopertha dominica, Cryptolestes ferrugineus, and Tribolium castaneum were scanned in the 1000–1600 nm wavelength range using an NIR hyperspectral imaging system. Dimensionality of the acquired hyperspectral data was reduced using multivariate image analysis. Six statistical image features (maximum, minimum, mean, median, standard deviation, and variance) and 10 histogram features were extracted from images at 1101.69 and 1305.05 nm and given as input to statistical discriminant classifiers (linear, quadratic, and Mahalanobis) for classification. Linear discriminant analysis and quadratic discriminant analysis classifiers correctly classified 85–100% healthy and insect-damaged wheat kernels.     

Detection and prediction of Sitophilus oryzae infestations in triticale via visible and near-infrared spectral signatures

Triticale is a hybrid of wheat and rye grown for use as animal feed. In Florida, due to its soft coat, triticale is highly vulnerable to Sitophilus oryzae L. (rice weevil) and there is interest in development of methods to detect early-instar larvae so that infestations can be targeted before they become economically damaging. The objective of this study was to develop prediction models of the infestation degree for triticale seed infested with rice weevils of different growth stages. Spectral signatures were tested as a method to detect rice weevils in triticale seed. Groups of seeds at 11 different levels (degrees) of infestation, 0–62%, were obtained by combining different ratios of infested and uninfested seeds. A spectrophotometer was used to measure reflectance between 400 and 2500 nm wavelength for seeds that had been infested at different levels with six different growth stages from egg to adult. The reflectance data were analyzed by several generalized linear regression and classification methods. Different degrees of infestation were particularly well correlated with reflectances in the 400–409 nm range and other wavelengths up to 967 nm, although later growth stages could be detected more accurately than early infestation. Stepwise variable selection produced the lowest mean square differences and yielded a high R² value (0.988) for the 4th instars, pupae and adults inside the seed. Models were developed to predict the level of infestation in triticale by rice weevils of different growth stages. Overall, this study showed a great potential of using reflectance spectral signatures for detection of the level of infestation of triticale seed by rice weevils of different growth stages.     

Fusarium damage assessment in wheat kernels by Vis/NIR hyperspectral imaging

Fusarium head blight is a fungal disease that affects the world’s small grains, such as wheat and barley. Attacking the spikelets during development, the fungus causes a reduction of yield and grain of poorer processing quality. Secondary metabolites that often accompany the fungus, such as deoxynivalenol (DON), are health concerns to humans and livestock. Conventional grain inspection procedures for Fusarium damage are heavily reliant on human visual analysis. As an inspection alternative, a near-infrared (NIR) hyperspectral image system (1000–1700 nm) was fabricated and applied to Fusarium-damaged kernel recognition. An existing extended visible (400–1000 nm) system was similarly used. Exhaustive searches were performed on the 144 and 125 wavelength pair images that, respectively, comprised the NIR and visible systems to determine accuracy of classification using a linear discriminant analysis (LDA) classifier. On a limited set of wheat samples the best wavelength pairs, either with visible or NIR wavelengths, were able to discriminate Fusarium-damaged kernels from sound kernels, both based on visual assessment, at an average accuracy of approximately 95%. Accuracy dropped off substantially when the visual contrast between the two kernel conditions became imperceptible. The NIR region was slightly better than the visible region in its broader array of acceptable wavelength pairs. Further, the region of interest (ROI) defined as the whole kernel was slightly better than ROIs limited to either a portion of the endosperm or the germ tip. For the NIR region, the spectral absorption near 1200 nm, attributed to ergosterol (a primary constituent in fungi cell membranes), was shown to be useful in spectral recognition of Fusarium damage.