紫色水稻土水稳性团聚体土壤微生物基因组DNA提取方法探讨

为了找到提取土壤微生物总DNA的最佳方法,通过OD值检验、凝胶电泳、PCR和DGGE分析,比较了Reddy法、基于DNAout kit试剂盒改进的实验方法、以及Kuske修订法、Edgcomb改进法、SDS高盐提取法、Eichner调整法等常用的不同土壤微生物基因组的DNA提取方法在亚热带地区长期免耕紫色水稻土水稳性团聚体0.25～2.0 mm粒径上的提取效果.结果表明,6种方法都可以从团聚体中提取到长度大于23.1 kb的DNA片段,但不同方法提取的DNA的产量存在明显差异,土壤总DNA均不需纯化就可以用于PCR扩增,使用细菌16S rDNA基因V3区的通用引物可扩增得到相应的片段.研究表明,改进的DNAout kit试剂盒法是长期免耕紫色水稻土水稳性团聚体中微生物基因组DNA的最佳提取方法.     

Effect of Micronization of Maize on Quality Characteristics of Pasta

Currently, production of pasta that is either gluten‐free or having lower content of gluten, using low‐cost nonwheat cereals and legumes, is becoming increasingly popular worldwide. This is mainly done to increase the nutritional value and reduce the allergenicity of the product. The quality attributes of pasta prepared from micronized maize flour with additives such as guar gum (MPG) and a combination of guar and xanthan gum (MPGX) were compared with pasta prepared from unmicronized flour with guar gum (UMPG). The optimum cooking time for pasta in all three cases (UMPG, MPG, and MPGX) was 3 min. The cooked weight of pasta MPG and MPGX was less compared with UMPG, indicating limited water penetration during cooking. The solid loss of pasta ranged between 8 and 9.5% and was within acceptable levels (<12%). Micronization increased the firmness in MPG (3.7 N) and MPGX (4.5 N) compared with UMPG pasta (2.7 N). MPGX pasta exhibited improved texture, color, and overall acceptability compared with UMPG, and these quality attributes were also comparable to commercial wheat pasta. The study indicated that micronized maize flour with gums can be used in the preparation of maize pasta with good quality attributes.     

Effect of Hydrophilic Gums on Frozen Dough. I. Dough Quality

Disadvantages of frozen doughs are their variable performance and loss of stability over long‐term frozen storage. Changes in rheological properties of frozen doughs have been reported to be due to the physical damage of the gluten network caused by ice crystallization and recrystallization. The objective of this study was to determine the effect of hydrophilic gums on ice crystallization and recrystallization for improvement of the shelf‐life stability of frozen dough. The present research involved use of the Hard Red Spring wheat cultivar Grandin and hydrophilic gums such as carboxymethyl cellulose (CMC), gum arabic, kappa carrageenan (κ‐carrageenan), and locust bean gum at three different levels each on doughs stored frozen for up to 16 weeks. The dough characteristics were analyzed after day 0, day 1, and after 4, 8, 12, and 16 weeks of frozen storage using data from differential scanning calorimetry (DSC), water activity, extensigraph, and proof time. The ΔH value of freezable water endothermic transitions obtained using DSC increased with storage time for all treatments. However, addition of different levels of the four gums lowered the ΔH value, indicating a decrease in freezable water. Doughs with locust bean gum gave a higher peak force, measured using the Kieffer dough extensibility rig of the texture analyzer, and lower proof time, indicating better retention of baking quality. Maximum resistance to extension increased upon addition of 1 and 3%; CMC; 1 and 3%; κ‐carrageenan; and 1, 2, and 3% locust bean gum as compared with the control. The various periods of storage or gum treatments did not affect the water activity of the thawed frozen doughs. Doughs with locust bean gum gave significantly lower proof time compared with the other treatments and the control. CMC gave the second lowest values, followed by gum arabic treatment. Addition of κ‐carrageenan increased the proof time compared with the control. In summary, locust bean gum, gum arabic, and CMC improved the dough characteristics to varying degrees. κ‐Carrageenan was the only gum that showed a detrimental effect on frozen dough.     

Effect of Hydrophilic Gums on the Quality of Frozen Dough: Electron Microscopy,Protein Solubility,and Electrophoresis Studies

Hydrophilic gums have been shown to improve the shelf‐life stability of frozen doughs during long periods of frozen storage. The objective of this research was to determine the effect of gums on starch and protein characteristics of frozen doughs using electron microscopy and electrophoresis studies. Frozen doughs, supplemented with three levels of gum arabic, carboxy methyl cellulose (CMC), kappa (κ) carrageenan, and locust bean gum, were studied after day 1 and after 4, 8, 12, and 16 weeks of frozen storage. Changes in the ultra structure of the frozen doughs were investigated, as well as the solubilities and composition of dough proteins by SDS‐PAGE. Scanning electron micrographs of doughs evaluated on day 0 (unfrozen) showed starch granules securely embedded in the gluten matrix. However, after 8 and 16 weeks of frozen storage, the frozen control dough without the gum additives clearly showed damage to the gluten network, and the starch granules appeared to be separated from the gluten. Doughs with locust bean gum and gum arabic showed better retention of the gluten network compared with the frozen control evaluated after different periods of storage. The SDS‐soluble protein content increased while residue protein content decreased as the frozen storage time increased. After each frozen storage period, the control dough without the gum additive had the highest amount of SDS‐soluble proteins and the lowest amount of residue proteins when compared with the doughs treated with gums. κ‐Carrageenan and locust bean gum had the lowest amount of SDS‐soluble proteins compared with doughs with CMC and gum arabic. The frozen control had the lowest amount of residue proteins at any particular time of frozen storage. κ‐Carrageenan treated doughs had the highest amount of residue proteins, followed by doughs with locust bean gum. Doughs with gum arabic and CMC had the lowest amount of residue proteins but still higher than the control doughs.     

Differentiation of carbohydrate gums and mixtures using fourier transform infrared spectroscopy and chemometrics

Guar gum, a nonionic galactomannan, is used as an economical thickener and stabilizer in the food industry and is often combined with xanthan, locust bean gum (LBG), or carboxymethylcellulose (CMC) to promote synergistic changes in viscosity or gelling behavior via intermolecular interactions; however, the adulteration of LBG with guar gum is a well-known industrial problem. The ability to identify the purity of gums and concentrations of individual gums in mixtures would be advantageous for quality control in the food industry. Fourier transform infrared spectroscopy (FTIR) methods are rapid and require minimum sample preparation. The objectives of this study were to evaluate the ability of FTIR techniques to (1) differentiate LBG with a variety of mannose/galactose (M/G) ratios, (2) differentiate guar, LBG, tara, and fenugreek gums, (3) differentiate pure guar gum from guar gum mixed with LBG, xanthan gum, or CMC, (4) quantify LBG, xanthan gum, and CMC in guar gum, and (5) quantify guar gum in LBG. Two FTIR methods were used: diffuse reflectance (DRIFT) on powdered gum samples added to KBr at 5%, w/w, and attenuated total reflectance (ATR) on 1%, w/w, gum solutions. Spectra were collected and then analyzed by multivariate statistical procedures (chemometrics). The DRIFT method provided better discrimination and quantitative results than the ATR method. Canonical variate analysis (CVA) of DRIFT spectra (1200-700 cm(-1)) was able to classify LBG with various M/G ratios, pure galactomannans, and pure versus mixtures of gums with 100% accuracy. Quantification of an individual gum in gum mixtures (0.5-15%, w/w) was possible using partial least-squares (PLS) analysis of DRIFT spectra with R2 > 0.93 and using this approach for quantifying guar gum added to LBG resulted in an R2 > 0.99, RMSEC = 0.29, and RMSEP = 3.31. Therefore, the DRIFT FTIR method could be a useful analytical tool for quality control of select gums and gum mixtures used in the food industry.     

Modification of Starch by Dry Heating with Ionic Gums

Waxy maize (native and hydroxypropylated [HP]) and potato starches were impregnated with ionic gums (sodium alginate, CMC, and xanthan, 1% based on starch solids) and heat‐treated in a dry state for 0, 2, or 4 hr at 130°C. Effects of the dry heating on paste viscosity (RVA) and clarity (light transmittance) were examined. Heat treatment with sodium alginate and CMC raised the paste viscosities of native and HP waxy maize starches, but decreased that of potato starch. Xanthan provided the most substantial changes in paste viscosity among the tested gums. It appeared to heavily restrict granule swelling of the waxy maize starches, but it increased swelling of potato starch granules. Dry heating raised the paste viscosity of all the starch‐gum mixtures tested, except the potato starchalginate mixture. The final viscosity at 50°C of a 7% paste was raised in all other starches by ≈500–1,000 cP by this treatment. The paste of waxy maize starch‐gum products became opaque and shorter textured by the heat treatment, regardless of the gum type, whereas potato starch‐gum products did not show any obvious change in paste clarity. Ionic gums could behave as cross‐linking agents as well as form graft copolymers through heatinduced ester formation. This simple heating process with ionic gums could be used as a modification method for starch.     

Toward the recognition of structure-function relationships in galactomannans

In this paper the determination of the physical/rheological characteristics is described for a series of commercial galactomannans of which the structural details have been reported previously. Both solubility of the galactomannans and rheological properties of galactomannan solutions and galactomannan/xanthan mixtures were determined. Using a statistical analysis approach an attempt was undertaken to recognize correlations between structural and rheological data. The best correlation found was between the abundance of galactose substituents at a regular distance (type of galactomannan) and the storage modulus (G') of mixed galactomannan/xanthan gels, underscoring the hypothesis that branching hinders the formation of a network with xanthan gum. Also, the G' for the group of locust bean gums correlated with the degree of blockiness, that is, the size and occurrence of nonsubstituted regions on the mannose backbone. In addition, galactomannans displayed an apparent decrease in gelling ability with increasing average molecular weight. That G' also relates to the type of galactomannan can therefore partly be attributed to differences in average molecular weight for the various galactomannan types. However, within the series of locust bean gums tested, also an increase of G' with molecular weight was observed. This can be explained by the decreasing number of loose ends of the polymers and the concomitant increasing efficiency in network participation with increasing molecular weight.     

Network Formation by Pilot Plant and Laboratory‐Extracted Barley β‐Glucan and Its Rheological Properties in Aqueous Solutions

Barley and oat β‐glucans of low viscosity form reversible gels when prepared in sufficiently high concentrations. Solutions of three barley β‐glucan gums differing in molecular weight and thus in viscosity were prepared at 1.0, 2.5, or 5.0% (w/w) concentration levels. Medium‐ and high‐viscosity gums were prepared in a pilot plant (PP) and laboratory (LAB), respectively. Low‐viscosity (LV) gum was extracted in the laboratory at pH 7, which allowed for native enzymatic activity and decreased molecular weight. Network formation was monitored overnight through changes in storage (G′) and loss (G″) moduli. The strength of the formed network was determined from oscillatory rheological measurements by increasing the strain from 2 to 100%. Findings demonstrate that gelation of β‐glucan is molecular weight dependent and practically an instantaneous process for low‐viscosity gum solutions at concentrations of ≤5% gum (or ≤4% β‐glucan), levels lower than previously anticipated. The purity of β‐glucan also seems to affect gelation rate. Better understanding of the β‐glucan gelation behavior is important for its functionality in both food product applications and physiological mechanisms of its health benefits.     

Characterization of Water State in Masa with Different Additives

Carboxymethyl cellulose (CMC) is added to tortillas to maintain a pliable texture during storage. A need exists to optimize or replace CMC in masa and tortilla manufacturing with cheaper yet adequate alternatives. Change in water distribution upon gum addition may be key to understanding stability of cooked masa. Therefore, the objective of this study was to characterize the state and distribution of water in masa systems containing two types of CMC and guar gum. Masa was mixed with 10% (1% in viscosity measurements) of different gums (either one of two CMCs varying in viscosity or guar) then hydrated to 50% moisture content. Viscosity, water holding capacity (WHC), total moisture content (TGA) as well as “freezable” (FW) and “unfreezable” (UFW) water (DSC) of all samples were obtained and compared. Viscosity measurements indicated guar gum may provide a good substitution for high viscosity CMC. The two water measurements, WHC and UFW, differed as to the effect of viscosity on water entrapment. WHC represented the short‐term imbibing of gums, while UFW indicated how the hydrocolloids responded in masa given full hydration time. UFW in guar gum was lower than in medium viscosity CMC. These initial results indicate that guar gum may prove a good substitute for CMC in masa applications.     

Remediation of a diesel-contaminated soil from a pipeline accidental spill: enhanced biodegradation and soil washing processes using natural gums and surfactants

Purpose

This paper addresses the application of bioproducts produced by plants (locust bean, guar, and mesquite seed gums) to enhance remediation processes of different nature: soil washing and biodegradation methodologies.

Materials and methods

These natural gums were tested at laboratory scale to remove total petroleum hydrocarbons-diesel fraction (TPH-diesel) from oil-contaminated volcanic soils sampled from a polluted site in an agricultural area of western Mexico. TPH-diesel removal by natural gums was compared to common synthetic surfactants.

Results and discussion

There is a strong evidence of contamination caused by the presence of TPH-diesel at a concentration of 32,100 mg/kg, which is above the legal limit of 1,200 mg/kg for agricultural soils in Mexico. Regarding the surfactant soil washing experiments, ionic surfactants showed removal rates above the control test of about 78.51 % (Maranil LAB), 71.27 % (Texapon 40), 60.13 % (SDS), and 48.19 % (Surfacpol G). In contrast, some nonionic surfactants showed removal rates below soil-washing background rate (40 %). On the other hand, natural gums showed interesting and promising results. Guar gum and locust bean gum showed efficiencies of 54.38 % and 53.46 %, respectively. Biodegradation experiments confirmed the effectiveness of natural gums as biodegradation enhancers in diesel-contaminated soils. Specifically, guar gum showed an excellent performance. An 82 % TPH-diesel removal rate was achieved for a very low gum concentration (2 ppm). In this particular context, reported surfactant concentrations to assist biodegradation are, in general, higher.

Conclusions

This work demonstrated the applicability of natural gums as soil remediation enhancers in diesel-contaminated systems. Particularly, guar gum might represent a cost-effective alternative for biodegradation enhancement processes.     

Application of a real time polymerase chain reaction method to detect castor toxin contamination in fluid milk and eggs

The castor seed contains ricin, which is one of the most potent biological toxins and is widely considered to be a threat agent for bioterrorism. In this study, a rapid and sensitive PCR method was applied to the detection of castor contamination in milk and liquid egg samples. The targeting gene sequence of the primer set, Ricin-F4/R4, was not found in either the bovine or chicken genome. Primers against a highly conserved sequence from the 18S ribosomal RNA gene were used as a positive control for DNA extraction and PCR reaction efficiency. The quantity and quality of DNA prepared from castor spiked or nonspiked milk and egg samples obtained from three different DNA extraction methods were compared. The cetyl trimethylammonium bromide (CTAB) method yielded the highest quality of DNA and is most suitable for the sensitive detection of castor DNA by real-time PCR in both milk and liquid egg matrixes. However, taking time and cost into consideration, a commercial kit designed for extraction of DNA from stool samples could be used as an alternative method for the routine extraction of DNA from milk for real-time PCR assays. The egg matrix was found to inhibit PCR amplification and interfere with two of the three methods tested for DNA extraction. Egg yolk had a greater negative effect on PCR amplification than the egg white matrix. Our results affirm the necessity of performing individual validations for each food matrix. Both real-time PCR systems used in this study, TaqMan and SYBR Green I dye, were capable of detecting 100 ng of castor acetone powder, corresponding to 5 ng of ricin, in 1 mL of milk or liquid egg, well below the toxic dose for humans. On the basis of these results, the real-time PCR method for detection of intentional castor contamination is applicable to milk and egg matrixes.     

Hydrocolloid interaction with water, protein, and starch in wheat dough

Interaction of hydrocolloids (xanthan gum, locust bean gum, guar gum, and high-methoxyl pectin) with macrocomponents of dough (water, starch, and protein) was evaluated by different techniques. (1)H spin-spin NMR relaxation assays were applied to study the mobility of the gluten-hydrocolloid-water matrix, and the amount of freezable water was determined by differential scanning calorimetry (DSC). Starch gelatinization parameters (T, enthalpy) were also analyzed by DSC. The influence of additives on the protein matrix was studied by Fourier transform (FT) Raman assays; analysis of the extracted gliadins and glutenins was performed by electrophoresis (SDS-PAGE). A significantly higher molecular mobility was found in matrices containing xanthan gum, whereas pectin led to the lowest molecular mobility. Freezable water showed a trend of increasing in the presence of hydrocolloids, particularly under conditions of water restriction. Starch gelatinization final temperature was decreased when hydrocolloids were added in the presence of enough water. In general, FT-Raman and SDS-PAGE indicated that hydrocolloid addition promoted a more disordered and labile network, particularly in the case of pectin addition. On the other hand, results obtained for dough with guar gum would indicate a good compatibility between this hydrocolloid and the gluten network.     

Identification of PCR-amplified genetically modified organisms (GMOs) DNA by peptide nucleic acid (PNA) probes in anion-exchange chromatographic analysis

PCR products obtained by selective amplification of transgenic DNA derived from food samples containing Roundup Ready soybean or Bt-176 maize have been analyzed by anion-exchange HPLC. Peptide nucleic acids (PNAs), oligonucleotide analogues known to bind to complementary single-stranded DNA with high affinity and specificity, have been used as specific probes in order to assess the identity of the peaks observed. Two different protocols were adopted in order to obtain single-stranded DNA: amplification with an excess of one primer or digestion of one DNA strand. The single-stranded DNA was mixed with the PNA probe, and the presence of a specific sequence was revealed through detection of the corresponding PNA:DNA peak with significantly different retention time. Advantages and limits of this approach are discussed. The method was tested with reference materials and subsequently applied to commercial samples.     

DIVERSITY AND DYNAMICS OF MANAGEMENT OF GUM AND RESIN RESOURCES IN ETHIOPIA: A TRADE‐OFF BETWEEN DOMESTICATION AND DEGRADATION

Although the human domestication of forest and tree resources is often considered to result in resource degradation, it may also lead to improved resource potentials. This paper assesses the nature and dynamics of gum and resin focused woodland exploitation and management systems in Ethiopia in the context of degradation and domestication processes. In three sites with commercial gum resin producing woodlands and production history, we studied variation in (i) woodland management and gum resin production systems and (ii) socio‐economic and biophysical factors that condition the management and production systems. On the basis of their organizational features, we formulated nine production models and related them to different phases of domestication and different degrees of ecosystem degradation. The production systems gradually evolved from the extraction of wild trees to production in an adapted forest system. However, domesticated woodlands with an adapted forest structure and composition and increased provisioning services are still little developed despite decades of production history. Many of these woodlands are undergoing serious degradation because of low quality management practices. This is mainly attributable to existing land use practices and the social arrangements for the production of and trade in the gums and resins. The findings illustrate that domestication involves not only a change in ecological and production systems but also the development of social arrangements for production and trade. We conclude that the status of domestication in a social sense determines whether forests and/or specific forest resources are degraded or aggraded in the sense of resource enrichment. Copyright © 2011 John Wiley & Sons, Ltd.     

一步双重PCR法检测梨火疫病原细菌（Erwinia amylovora）

由格兰氏阴性菌 Erwinia amylovora引起的梨火疫病对梨树、苹果树以及一些观赏性植物具有极大的破坏性。研究表明自然界中存在不含有pEA29质粒的E. amylovora菌株,基于质粒pEA29序列设计的分子检测方法则不能检测出该菌株。本研究采用两对引物,分别来自染色体和pEA29质粒,同时在一个反应体系中进行PCR反应,扩增片段长度分别为1.6 kb和1.0 kb,可特异性检测不含有pEA29质粒的菌株。该方法可以简单、快速、准确地检测梨火疫病原细菌。     

步双重PCR法检测梨火疫病原细菌(Erwinia amylovora)

由梨火疫病原细菌（Erwinia amylovora）导致的火疫病（fire blight）是梨、苹果及其它蔷薇科植物上的毁灭性病害,我国将其定为对外一类检疫性有害生物。该病随种苗、果实及包装材料传播。目前国际上所建立的E.amylovora分子检测技术主要基于该病菌所含有的pEA29质粒序列,研究发现失去质粒的人工突变菌株的致病力下降，认为质粒对于梨火疫病菌的致病力是至关重要的。虽然带有pEA29质粒的梨火疫病菌菌株在自然界中占有绝对的数量，但Llop et al．（2006）报道了自然侵染的植物中存在具有致病力但不含pEA29质粒的梨火疫病菌（IVIA1614-2a菌株）。     

Rapid isolation of DNA from chocolate and date palm tree crops

DNA isolation from plants is sometimes difficult due to the existence of high levels of endogenous phenolics, polysaccharides, or other substances that may interfere with DNA extraction. Theobroma cacao produces high levels of anthocyanins in young leaves. These plant polyphenols can interfere with DNA isolation. After examination of various procedures for DNA isolation, two commercial isolation procedures have proved to be repeatedly successful using these types of plants, the D(2) BioTechnologies DNA X-tract Plus kit and the Qiagen DNeasy Plant System DNA kit. These commercial kits were chosen for their speed and ease over the CTAB procedure, which is more labor intensive. All protocols assessed yielded DNA suitable for AFLP or SSR procedures. An additional factor in DNA extraction efficiency is the degree of cell breakage, which may be more difficult with the highly fibrous leaf tissue that is found in many monocots, including date palm. Two commercially produced pieces of equipment were tested and, for cacao, both resulted in template DNA yielding amplification product in AFLP or SSR fingerprinting. However, for the fibrous date palm leaf, the larger FastPrep homogenizer consistently yielded DNA that generated higher signals when amplified than did the smaller Disruptor Genie.     

Validation of a tRNA-Glu-cytochrome b key for the molecular identification of 12 hake species (Merluccius spp.) and Atlantic Cod (Gadus morhua) using PCR-RFLPs, FINS, and BLAST

The goal of this study was to develop a diagnostic key for hake meat to solve the limitations of previous identification methodologies, mainly related to the high degradation of the DNA recovered from processed foods. We describe the development of two molecular tools based on polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphisms of the cytochrome b gene, respectively, to identify DNA from 12 hake species in commercial products. The first assay is an exclusion test consisting of the PCR amplification of a 122 bp fragment using nested primers interspecifically conserved in Merluccius spp. and in Gadus morhua. This 122 bp amplicon, being the shortest one so far designed for hake DNA, is a useful traceability tool for highly degraded samples because its sequence contains enough interspecific diagnostic variation to identify 10 hake species and cod and has been successfully amplified from most commercial products so far tested. The second identification key follows a positive outcome of the exclusion test and consists of the PCR amplification of a 464-465 bp fragment and its digestion with three restriction enzymes whose targets map at interspecifically nonconserved sites of the cytochrome b. The key presented here has passed through a rigorous methodological calibration including its testing for genus specificity, its validation on a large number of authenticated sample types from each species range, and its implementation with a maximum likelihood method for the assignment of unknown samples. Together, these two procedures constitute the most complete molecular key so far developed for Merluccius spp., which is optimal for routine identification of hakes in large commercial samples at a reasonable cost-time ratio.     

Identification of the razor clam species Ensis arcuatus, E. siliqua, E. directus, E. macha, and Solen marginatus using PCR-RFLP analysis of the 5S rDNA region

Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis of the 5S ribosomal DNA region has been applied to the establishment of DNA-based molecular markers for the identification of five razor clam species: Ensis arcuatus, E. siliqua, E. directus, E. macha, and Solen marginatus. PCR amplifications were carried out using a pair of universal primers from the coding region of 5S rDNA. S. marginatus was simply distinguished by the different size of the amplicons obtained. Species-specific restriction endonuclease patterns were found with the enzymes Hae III for E. arcuatus, E. siliqua, and E. directus, and Acs I for E. macha, and when two enzymes were combined, the four species were also identified. Thus, this work provides a simple, reliable, and rapid protocol for the accurate identification of Ensis and Solen species in fresh and canned products, which is very useful for traceability and to enforce labeling regulations.     

Differential detection of shrimp and crab for food labeling using polymerase chain reaction

Shrimp and crab are well-known as allergenic ingredients. According to Japanese food allergy labeling regulations, shrimp species (including prawns, crayfishes, and lobsters) and crab species must be differentially declared when ≥10 ppm (total protein) of an allergenic ingredient is present. However, the commercial ELISA tests for the detection of crustacean proteins cannot differentiate between shrimp and crab. Therefore, two methods were developed to discriminate shrimp and crab: a shrimp-PCR method with postamplification digestion and a crab-PCR method that specifically amplifies a fragment of the 16S rRNA gene. The sensitivity and specificity of both PCR methods were verified by experiments using DNA extracted from 15 shrimp species, 13 crab species, krill, mysid, mantis shrimp, other food samples (cephalopod, shellfish, and fish), incurred foods, and commercial food products. Both PCR methods could detect 5 pg of DNA extracted from target species and 50 ng of genomic DNA extracted from incurred foods containing 10 ppm (μg/g) total protein of shrimp or crab. The two PCR methods were considered to be specific enough to separately detect species belonging to shrimp and crab. Although false-positive and false-negative results were obtained from some nontarget crustacean species, the proposed PCR methods, when used in conjunction with ELISA tests, would be a useful tool for confirmation of the validity of food allergy labeling and management of processed food safety for allergic patients.