Thin layer chromatographic method for the detection of uric acid: collaborative study

A collaborative study has been completed on an improved method for the detection and confirmation of uric acid from bird and insect excreta. The proposed method involves the lithium carbonate solubilization of the suspect excreta material, followed by butanol-methanol-water-acetic acid thin layer chromatography, and trisodium phosphate-phosphotungstic acid color development. The collaborative tests resulted in 100% detection of uric acid standard at the 50 ng level and 75% detection at the 20-25 ng level. No false positives were reported during tests of compounds similar to uric acid. The proposed method has been adopted official first action; the present official final action method, 44.161, will be retained for screening purposes.     

Flotation technique for quantitative determination of mite populations in powdered and compacted foodstuffs.

Samples are preheated in 25% aqueous ethanol at 65 degrees C for 5 min. A specially designed flotation flask and solvent mixture of kerosene-tetralin (1+1) is used to form an interface with 25% aqueous ethanol. Mites, insects, and insect fragments are separated from vegetative material by an overflow method. Trials incorporating a known number of mites in samples of pigmeal indicate that at an infestation level of 1000 mites/kg the method always recovered 70% of the population and at 2000 mites/kg, 75%. The method permits collection of mite specimens for microscopic examination as well as allowing differentiation between live and long-dead insects. It does not use highly volatile solvents or corrosive acids. The apparatus has been successfully tested for quantitatively determining mite populations in flour, dried skimmed milk, animal feedstuffs, soya meal, wheat, rape seed, dates, and house dust. It has also been used to estimate insect contamination in packaged dates.     

Extraction of light filth from ground nutmeg: collaborative study

The official method for extracting light filth from ground nutmeg, 44.116, gives variable recoveries of filth elements and results in large amounts of interfering plant material in the light filth extraction. A new method has been developed using a chloroform pretreatment for heavy filth followed by a water-40% isopropanol-mineral oil flotation of light filth. A comparison of the methods in the collaborative study showed a higher recovery of insect fragments, 90.6 vs. 65.2%, and of rodent hairs, 86.5 vs. 66.5%, for the new method over the present official first action method, 44.116. Frothing, an additional problem with the official method, has been reduced by the new method. The new method has been adopted as official first action to replace the existing method.     

Microwave drying for safe storage and improved nutritional quality of green gram seed (Vigna radiata)

The present study describes the effect of the microwave-heating method on disinfestations and physico characteristics, viz., grain size, grain hardness, and nutritional quality, of the stored green gram seed. It has been observed that the use of the microwave-heating method not only prolongs the storage duration of the green gram seed but also enhances its nutritional quality. The effect of independent parameters, viz., microwave power level and time of exposure, on the moisture content, insect mortality, color, and antinutrient factor (phyic acid) was optimized using response surface methodology (RSM), with the optimized value for power of 808 W and time at 80 s. The optimally treated green gram seed has 8.9% moisture, 99.5% insect mortality, 2.22 Δa* (green color of seed), and 591.79 mg/100 g of antinutrient factor (phytic acid). The grain size (geometric mean diameter, D(m)) of the control (untreated) sample was 3.75 mm, and that of the microwave-treated sample using optimum conditions was 3.99 mm. The grain hardness of the control sample was 3.31 kg, and that of the microwave-treated sample using optimum conditions was 1.305 kg. In vitro protein digestibility (IVPD) of the control (untreated) sample was 83 ± 0.289%, and that of the microwave-treated sample using optimum conditions was 85 ± 0.296%. These values are significantly difference (p < 0.05). The mineral elements studied were Zn, Fe, Mg, Mn, Cu, K, Ca, and Na. The microwave treatment resulted in a non-significant (p < 0.05) effect for Mg, Mn, Cu, K, and Na but a significant (p < 0.05) effect for Zn, Ca, and Fe. The results indicate that the microwave heating not only increases the insect mortality but also reduces the moisture content and antinutritional factor (phytic acid), while the natural green color of the seed is not affected much. This study provides a novel and environmentally safe technique and increase in the nutritive quality.     

Extraction of light filth from spirulina powders and tablets: collaborative study

Results are reported for a collaborative study of a method for the extraction of light filth from spirulina (a blue-green alga) powder and tablets. A 50 g portion of either powder or tablets is dispersed in water, and then boiled with dilute HCI solution. Hairs and insect fragments are isolated by wet sieving on a No. 230 sieve, flotation with mineral oil, and washings of the mineral oil in a percolator. Average recoveries by 12 collaborators for tablets and powders were 70.6 and 70.2%, respectively, for 10 rat hair spikes and 68.3 and 84.4%, respectively, for 20 insect fragment spikes. The method has been approved interim official first action.     

Analysis of natural coloring matters in food. III. Application of methylation with diazomethane for the detection of lac color

A new method has been proposed for detection of lac color in food. Lac color is a natural color additive derived from a secretion of the insect Coccus Laccae (Laccifer lacca Kerr). It is extracted from food with methanolic oxalic acid and eluted from a column of Amberlite XAD-2 with the same solvent. The fraction containing the lac color is treated with diazomethane to produce 2 reddish-orange markers. The marker species in the reaction mixture are detected by both thin-layer chromatography and reverse-phase liquid chromatography.     

Extraction of light filth from coconut: collaborative study

The official method for filth in coconut, 44.029, frequently produces filter papers with excessive plant debris and low hair recoveries. To overcome these deficiencies, 3 changes in the method were made: (1) the defatting step was modified to use sodium lauryl sulfate in combination with borax for better defatting; (2) 40% isopropanol was substituted for 60% ethanol as the aqueous phase for cleaner papers; and (3) mineral oil was substituted for n-heptane to improve hair recoveries. These changes resulted in higher, more reproducible recoveries of rodent hairs and insect fragments and cleaner filter papers. The method has been adopted as official first action to replace 44.029.     

Improved spray reagent for thin layer chromatographic method for detecting uric acid: collaborative study

A collaborative study was conducted to validate the substitution of an improved single spray in the official AOAC thin layer chromatographic method for identifying uric acid (UA) from bird and insect excreta. The proposed reagent, which is a dilute aqueous solution of ferric chloride and potassium ferricyanide, requires neither a heating step nor a pH indicator. Its preparation time, specificity, and sensitivity to low levels (5-50 ng) of UA were compared with those of the official sprays. The improved spray took 1/5 as long to prepare as the official sprays. Neither the proposed spray nor the official sprays gave false positive reactions with compounds similar to UA. For bird and insect excreta samples, at the 95% confidence limits, the false negative rate was between 0 and 9.7% for the proposed spray and between 0.7 and 18.7% for the official sprays. Sensitivity results showed that the proportion positive for the proposed spray was significantly higher (P less than 0.05) than for the official sprays at the 15 ng UA level. The proposed changes have been adopted official first action.     

Extraction of light filth from whole peppermint leaves: collaborative study

Results are reported for a collaborative study of a method for the extraction of light filth from whole peppermint leaves. A 5 g sample is defatted with isopropanol in a simple reflux appartus. Rat hairs, insect fragments, and whole insects are isolated by wet sieving on a No. 230 sieve, a deaerating boil in 40% isopropanol solution, flotation with Tween 80-Na4EDTA (1 + 1) and mineral oil-heptane (85 + 15), and trappings in a Wildman trap flask. Average recoveries obtained by 6 collaborators for 3 spike levels of rat hairs (5, 10, 15) were 83.3, 87.5, and 82.2%, respectively. For whole insects (5, 10, 15) recoveries averaged 85.0, 80.0 and 77.2% respectively; for insect fragments (20, 30, 50) recoveries averaged 79.6, 88.3, and 84.8%, respectively. The average recoveries for the 3 levels of each analyte were not significantly different. The method has been adopted official first action.     

Extraction of light filth from unground marjoram: collaborative study

The present method for filth in underground marjoram is conducted in 2 parts. The first of these, which is for heavy filth and sand, requires the spice to be boiled with petroleum ether, then floated off with chloroform and, if needed, carbon tetrachloride. The second part, which is for light filth, is dependent on completion of the heavy filth section. After the spice is air-dried, the light filth is extracted with heptane and water. The proposed method was developed to make light filth independent of heavy filth analysis, improve filth recoveries, and reduce microscopic examination time. The light filth is extracted by ethanol defatting followed by a combination 15-60% ethanol/mineral oil extraction in a Wildman trap flask. Tween 80-NaEDTA solution is added to the trapping system to reduce the rising of excess plant material to the trap interface. The official method (AOAC 13th edition secs 44.142, 44.120(b)) produced an average of 8 extraction papers per test portion and microscopic examination took an average of 88 min per test portion. The official method gave average recoveries of 14% for rodent hairs and 26% for insect fragments with 2 spike levels of 15 and 30 for each filth element. Recoveries of the 2 levels of each spike were not significantly different (P greater than 0.05). The proposed method gave average recoveries of 73% for rodent hairs and 70% for insect fragments. The proposed method has been adopted official first action to replace AOAC 13th edition secs 44.142 and 44.120(b) for unground marjoram only.     

基于改进DenseNet和迁移学习的荷叶病虫害识别模型

病虫害的发生将会严重影响莲藕品质与产量,开展病害诊断与识别对藕田病虫害及时对症对病诊治、提升莲藕生产质量与经济效益具有重要意义。该研究以荷叶病虫害高效、准确识别为目标,提出了一种基于改进DenseNet和迁移学习的荷叶病虫害识别模型。采用分支结构对模型的浅层特征提取模块进行改进,并在Dense Block与Transition Layer中引入Squeeze and Excitation注意力机制模块和锐化的余弦卷积,最后基于Plantvillage数据集进行迁移学习,实现了91.34%的识别准确率。该研究实现了对荷叶腐败病、病毒病、斜纹夜蛾、叶腐病、叶斑病的识别,并将改进后的模型推广应用于基于无人机图像的藕田病虫害检测,实现了病害分布可视化,可对莲藕病虫害的智能化防治提供有益指导。     

无人机航拍林业虫害图像分割复合梯度分水岭算法

针对林业信息监测方式实时性差、监测范围有限等问题,为更加实时、准确地对林业虫害信息进行监测并计算监测样地中虫害区域比例,该文在搭建面向林区虫害监测的多旋翼无人飞行器航拍监测系统基础上,提出了一种基于复合梯度分水岭算法的图像分割方法。该方法引入全局直方图均衡化消除了图像暗纹理的影响,并采用形态学混合开闭重构滤波完成了图像样本的去噪处理。计算灰度图像各像素点的复合梯度实现了非相关区域(道路及裸地)的提取,最终利用分水岭算法实现了监测图像虫害区域的分割提取。利用该文所提算法对8幅虫害侵蚀程度不同的监测图像进行分割,并与传统分水岭算法、K-means聚类算法进行对比试验。试验结果表明,该文算法虫害区域提取的平均相对误差率分别降低6.56%、3.17%,平均相对极限测量精度分别改善7.19%、2.41%,能够相对准确地将虫害区域从监测图像中分割出来,可为后续林业虫害监测与防护提供参考。     

Brine saturation technique for extraction of light filth from rubbed, ground, and whole sage: collaborative study

A new approach to the isolation of light filth from the 3 commercial forms of sage was studied collaboratively. It incorporates a simple isopropanol defatting, followed by saturation of the product with brine by alternately heating and cooling, and subsequent trapping of filth from tap water with olive oil. This method circumvents the use of hazardous, expensive solvents and more time-consuming pretreatment procedures. Overall recoveries were 92.1% for rodent hair and 78.7% for insect fragments on clean, easy-to-read papers. An additional blending step was necessary to obtain satisfactory recovery of rodent hair fragments from whole sage. The method has been adopted official first action for light filth in rubbed and ground sage only.     

Extraction of light filth from whole leaves of alfalfa, lemon balm, papaya, and spearmint: collaborative study

Results are reported for a collaborative study to extend AOAC method 44.A06-44.A08 to extraction of light filth from whole leaves of alfalfa, lemon balm, papaya, and spearmint. A 5 g (spearmint) or 10 g (alfalfa, lemon balm, papaya) test portion is defatted with isopropanol in a simple reflux apparatus. Rat hairs, insect fragments, and whole insects are isolated by wet sieving on a No. 230 sieve, a deaerating boil in 40% isopropanol, and flotation with mineral oil-heptane (85 + 15) from Tween 80-Na4EDTA (1 + 1) and 40% isopropanol in a Wildman trap flask. Each product was spiked at a different level. For rat hairs, recoveries averaged 82.2% from alfalfa, 88.9% from lemon balm, 80.6% from papaya, and 79.6% from spearmint. Recoveries of whole or equivalent insects from these products averaged 66.1, 218.8, 69.4, and 85.4%, respectively; recoveries of insect fragments from these products averaged 89.6, 94.4, 94.1, and 88.1%, respectively. The method has been adopted official first action for extraction of light filth from whole leaves of alfalfa, papaya, and spearmint. The extension of the method to lemon balm was not recommended because of interferences by intrinsic whole insects, which were the same species as the spike material.     

Detection of Fragments from Internal Insects in Wheat Samples Using a Laboratory Entoleter

A simple, rapid method that uses a small mechanical rotary device (entoleter) was developed for estimating insect fragment counts in flour caused by hidden, internal‐feeding insects in whole grains of hard red winter and soft red winter wheat. Known counts of preemergent adults, pupae, and larvae of lesser grain borers and rice weevils were blended with 500 g samples of uninfested wheat. The entoleter impeller speed was adjusted based on grain hardness and moisture content to obtain about ≈98% intact and ≈2–2.5% broken kernels in an uninfested sample. The entoleter flung the wheat kernels against a surrounding steel ring. Approximately 70–90% of the insect‐infested kernels, being weaker, released internal insect pieces upon impact. The broken kernels were sieved with number 10 and number 20 sieves to obtain large‐sieved and small‐sieved fractions, respectively. Insect pieces in sieved fractions were counted. The insect piece counts were correlated with the estimated flour fragments (R² = 0.94). The entoleter method can distinguish samples of grain containing 0, 25, or 75 fragments in 50 g of flour, with greater than 95% confidence. The method can be performed in approximately 5 min per 500 g sample and could potentially be a cost‐effective method that grain handlers can use to inspect wheat loads for detecting insect damage and estimating insect fragments in flour.     

Collaborative study of the determination of available lysine in proteins and feeds.

In the proposed method 1-fluoro-2,4-dinitrobenzene (DNFB) is reacted with the free epsilon-amino groups in protein of form DNFB-epsilon-amino lysine which is stable to acid hydrolysis. The sample is acid hydrolyzed and unavailable lysine is determined with an amino acid analyzer; total lysine is determined on the untreated sample. The available lysine, which was bound by DNFB, is determined by difference. The available lysine has been determined in 3 samples of 44% protein soybean meal by 5 collaborators, following the method outlined. The range for available lysine in reference standard 1 was 2.02-2.14%, in reference standard 2, 2.59-2.73% and in reference standard 3, 0.55-0.91%. The method has been adopted as official first action.     

Determination of spinosad and its metabolites in food and environmental matrices. 2. Liquid chomatography-mass spectrometry

A selective and sensitive method utilizing liquid chromatography-mass spectrometry (LC-MS) has been developed for determining residues of the natural insect control agent spinosad in several crop matrices that are difficult to analyze by HPLC with UV detection. The method determines the active ingredients (spinosyns A and D) and three minor metabolites (spinosyns B and K and N-demethylspinosyn D) in alfalfa hay, wheat hay, wheat straw, sorghum fodder, and corn stover. The analytes are extracted from the samples with an acetonitrile/water solution, and the extracts are purified by solid phase extraction with a C(18) disk and a silica cartridge. All five analytes are determined simultaneously in a single injection using positive atmospheric pressure chemical ionization LC-MS with selected ion monitoring. The average recoveries ranged from 69 to 96% with standard deviations ranging from 4 to 15%. The method has a validated limit of quantitation of 0. 01 microgram/g and a limit of detection of 0.003 microgram/g. The LC-MS method can also provide residue confirmation in addition to quantitation.     

Concise preparation of the (3E,5Z)-alkadienyl system. New approach to the synthesis of principal insect sex pheromone constituents

A new rapid and low-cost preparation of the (3E,5Z)-3,5-alkadienyl system, encountered in several insect pheromone constituents, was developed. Knoevenagel condensation of (E)-2-alkenals with ethyl hydrogen malonate in dimethyl sulfoxide, in the presence of a catalytic amount of piperidinium acetate, led to a mixture of geometrical isomers of ethyl 3,5-alkadienoates and ethyl 2,4-alkadienoates, from which the (3E,5Z)-3,5-alkadienoate was conveniently separated, by the use of urea inclusion complex formation. The importance of this procedure has been illustrated by the preparation of the (3E,5Z)-3,5-tetradecadienoic acid (megatomoic acid) 1, the (3E,5Z)-3,5-dodecadienyl acetate 2, and the (3E,5Z)-3,5-tetradecadienyl acetate 3. These compounds are the main components of insect sex pheromones and constitute synthetic targets of considerable interest for the semiochemical community.     

基于稀疏编码金字塔模型的农田害虫图像识别

相较于一般物体的图像,农作物害虫图像因具有复杂的农田环境背景,分类与识别更加困难。为提高害虫图像识别的准确率,该文提出一种基于图像稀疏编码与空间金字塔模型相结合的害虫图像表示与识别方法。该方法利用大量非标注的自然图像块构造过完备学习字典,并运用该学习字典实现对害虫图像的多空间稀疏表示。与此同时,结合多核学习,该文设计了一种害虫图像识别算法。通过对35种害虫的识别,试验结果表明：在相同方法下,该文所提特征提取方法可使平均识别精度提高9.5百分点;此外,进一步通过对221种昆虫及20种蝴蝶的识别,试验结果表明：与传统方法相比较,该文所提方法使得平均识别精度提高14.1百分点。     

防虫网对节能日光温室通风性能影响的研究

自然通风是决定日光节能温室质能平衡和调节微生态环境平衡的重要因素。为了提高节能日光温室的环境管理和结构设计水平,有必要研究防虫网对日光温室通风性能的影响,以提高日光温室的绿色产品生产率。因此,运用气体衰减示踪技术和数理统计理论,研究了日光温室不同形式通风机构的通风性能。试验研究表明在通风口处装设防虫网会显著地减少日光温室的通风率,有时可高达50%。研究还表明,无论装设有无防虫网,卷膜通风机构比拉膜(推膜)通风机构都具有较大的通风率,有时前者是后者的1.4倍左右。