Determination of the variability of both hydrophilic and lipophilic toxins in endemic wild bivalves and carnivorous gastropods from the Southern part of Chile

The aim of this study was to analyse and determine the composition of paralytic shellfish poisoning (PSP) toxins and lipophilic toxins in the Region of Aysén, Chile, in wild endemic mussels (Mytilus chilensis, Venus antiqua, Aulacomya ater, Choromytilus chorus, Tagelus dombeii and Gari solida) and in two endemic carnivorous molluscs species (Concholepas concholepas and Argobuccinum ranelliforme). PSP-toxin contents were determined by using HPLC with fluorescence detection, while lipophilic toxins were determined by using LC-MS/MS. Mean concentrations for the total of PSP toxins were in the range 55–2505 μg saxitoxin-equivalent/100 g. The two most contaminated samples for PSP toxicity were bivalve Gari solida and carnivorous Argobuccinum ranelliforme with 2505 ± 101 and 1850 ± 137 μg saxitoxin-equivalent/100 g, respectively (p < 0.05). The lipophilic toxins identified were okadaic acid, dinophysistoxin-1 (DTX-1), azaspiracid-1 (AZA-1), pectenotoxin-2 (PTX-2) and yessotoxins (YTX). All analysed molluscs contained lipophilic toxins at levels ranging from 56 ± 4.8 to 156.1 ± 8.2 μg of okadaic acid-equivalent/kg shellfish together with YTX at levels ranging from 1.0 ± 0.1 to 18 ± 0.9 μg of YTX-equivalent/kg shellfish and AZA at levels ranging from 3.6 ± 0.2 to 31 ± 2.1 μg of AZA-equivalent/kg shellfish. Furthermore, different bivalves and gastropods differ in their capacity of retention of lipophilic toxins, as shown by the determination of their respective lipophilic toxins levels. In all the evaluated species, the presence of lipophilic toxins associated with biotransformation in molluscs and carnivorous gastropods was not identified, in contrast to the identification of PSP toxins, where the profiles identified in the different species are directly related to biotransformation processes. Thus, this study provides evidence that the concentration of toxins in the food intake of the evaluated species (Bivalvia and Gastropoda class) determines the degree of bioaccumulation and biotransformation they will thereafter exhibit.     

Inter-species variability of okadaic acid group toxicity in relation to the content of fatty acids detected in different marine vectors

Okadaic acid group (OA-group) is a set of lipophilic toxins which are characterised by being produced by species associated with the genera Dinophysis and Prorocentrum. OA-group has been regularly detected in endemic shellfish species from the southern zone of Chile only through the mouse bioassay. The purpose of this work was to determine the variability of OA-group toxins in endemic aquatic organisms (bivalves, crabs, gastropods and fish) and to establish the relationship with the concentration of fatty acids (FAs) detected in the evaluated species. The toxicity of OA-group and the FA profiles were determined using LC-MS/MS and gas chromatography with flame-ionisation detection, respectively. In the study area, the dinoflagellate Dinophysis acuta was detected in densities ≈2000 cells ml^?1 with a toxicity ≈18.3 pg OA equiv cel^?1. The analysis identified OA and dinophysistoxin-1 in shellfish in a range of ≈90 to ≈225 μg OA eq kg^?1, where no toxins in fish were detected. A positive relationship between the FA level and the concentration of OA-group toxins in the digestive glands of bivalves and gastropods was established, noted for high levels of saturated FAs (C14:0 and C16:0). The toxic variability of OA-group toxins determined in the different species allowed us to establish that the consumption of these vectors, regulated by non-analytical methods, can be harmful when consumed by humans, thus suggesting that the sanitary regulations for the control of OA-group in Chile should be updated.     

Investigation of diarrhetic shellfish toxins in Lingshan Bay,Yellow Sea,China, using solid-phase adsorption toxin tracking (SPATT)

Early detection of toxin contamination in shellfish (i.e., prior to harvest) would be of considerable advantage to fish farmers, researchers and food safety administrators. In 2004, a solid-phase adsorption toxin tracking (SPATT) technique was developed to study algal toxins in New Zealand shellfish harvesting areas. In subsequent years, the basic idea have been further developed. Using a SPATT method, an investigation into diarrhetic shellfish toxins (DSTs) was conducted over a 10.5-month period in 2012 in shellfish farming areas in Lingshan Bay (Yellow Sea, China). This paper discusses the relationship among DSTs in toxic algae, seawater and contaminated shellfish. OA, DTX1 and PTX2 toxins were found in this shellfish farming area from summer to autumn. In shellfish the maximum concentrations of OA and DTX1 were 81 and 41 ng g^–1 respectively. PTX2 was very low. The maximum levels of OA and DTX1 in seawater were 165 and 56 ng g^–1 respectively, and were detected on June, separated by a 14-day period. Shellfish had accumulated the highest levels of OA and DTX1 recorded in this study. Comparison of the variations in DST levels in seawater showed there to be about 2 weeks for administrators to warn of the potential for toxin contamination in shellfish. Further research to explore the relationship between the variables of seawater temperature, sunlight and salinity, and DSTs in shellfish may help to establish a more suitable model for forecasting DST contamination in shellfish.     

Saxitoxins and okadaic acid group: accumulation and distribution in invertebrate marine vectors from Southern Chile

Harmful algae blooms (HABs) are the main source of marine toxins in the aquatic environment surrounding the austral fjords in Chile. Huichas Island (Aysén) has an history of HABs spanning more than 30 years, but there is limited investigation of the bioaccumulation of marine toxins in the bivalves and gastropods from the Region of Aysén. In this study, bivalves (Mytilus chilenses, Choromytilus chorus, Aulacomya ater, Gari solida, Tagelus dombeii and Venus antiqua) and carnivorous gastropods (Argobuccinum ranelliformes and Concholepas concholepas) were collected from 28 sites. Researchers analysed the accumulation of STX-group toxins using a LC with a derivatisation post column (LC-PCOX), while lipophilic toxins (OA-group, azapiracids, pectenotoxins and yessotoxins) were analysed using LC-MS/MS with electrospray ionisation (+/–) in visceral (hepatopancreas) and non-visceral tissues (mantle, adductor muscle, gills and foot). Levels of STX-group and OA-group toxins varied among individuals from the same site. Among all tissue samples, the highest concentrations of STX-group toxins were noted in the hepatopancreas in V. antiqua (95 ± 0.1 μg STX-eq 100 g^?1), T. dombeii (148 ± 1.4 μg STX-eq 100 g^?1) and G. solida (3232 ± 5.2 μg STX-eq 100 g^?1; p < 0.05); in the adductor muscle in M. chilensis (2495 ± 6.4 μg STX-eq 100 g^?1; p < 0.05) and in the foot in C. concholepas (81 ± 0.7 μg STX-eq 100 g^?1) and T. dombeii (114 ± 1.2 μg STX-eq 100 g^?1). The highest variability of toxins was detected in G. solida, where high levels of carbamate derivatives were identified (GTXs, neoSTX and STX). In addition to the detected hydrophilic toxins, OA-group toxins were detected (OA and DTX-1) with an average ratio of ≈1:1. The highest levels of OA-group toxins were in the foot of C. concholepas, with levels of 400.3 ± 3.6 μg OA eq kg^?1 (p < 0.05) and with a toxic profile composed of 90% OA. A wide range of OA-group toxins was detected in M. chilensis with a toxicity < 80 μg OA eq kg^?1, but with 74% of those toxins detected in the adductor muscle. In all evaluated species, there was no detection of lipophilic toxins associated with biotransformation in molluscs and carnivorous gastropods. In addition, the STX-group and OA-group toxin concentrations in shellfish was not associated with the presence of HAB. The ranking of toxin concentration in the tissues of most species was: digestive glands > mantle > adductor muscle for the STX-group toxins and foot > digestive gland for the OA-group toxins. These results gave a better understanding of the variability and compartmentalisation of STX-group and OA-group toxins in different bivalve and gastropod species from the south of Chile, and the analyses determined that tissues could play an important role in the biotransformation of STX-group toxins and the retention of OA-group toxins.     

Lipophilic toxin profiles detected in farmed and benthic mussels populations from the most relevant production zones in Southern Chile

Lipophilic toxins associated with diarrhoeic toxins were found in Mytilus chilensis (Blue mussels) and Aulacomya ater (Ribbed mussels). These shellfish samples were collected from Chiloe Island, Southern Chile. The samples were tested by liquid chromatography–tandem mass spectrometry (LC-MS/MS). After the analysis, four toxins were found: DTX-1, DTX-3, YTX and PTX. All toxins were identified by comparing their HPLC retention times with those of analytical standards and confirmed by LC-MS/MS. Dinophysistoxin-1 (DTX-1) and dinophysistoxin-3 (DTX-3) toxins were the major components within the mussel extracts. Nevertheless, the percentages of these toxins differed depending on the area they were collected from and/or the sampling date. The levels detected in Butacheuques Island for okadaic acid (OA) was 267?±?3.5?µg OA?eq?kg^?1 (p?<?0.05) and for DTX-3 was 183.4?±?7.5?µg?kg^?1 in ribbed mussels. Pectenotoxin (PTX) and yessotoxin (YTX) were the toxins detected in minor proportions in the toxic profile of the bivalves. The maximum concentration of YTX detected in ribbed mussels was 85.2?±?2.8?µg?kg^?1 in Mechuque Island, whereas the PTX-2 level in ribbed mussels was 82.0?±?2.4?µg?kg^?1 in Cailin Island. Analogues of YTX and PTX-2 were not detected in any of the analysed mussels, which did not support the supposed presence of isomers of toxins as a result of the enzymatic metabolism of bivalves. This study found evidence proving co-occurrence of lipophilic toxins – like PTX and YTX – with diarrhoeic toxin in samples collected in Southern Chile, which is, to date, the more complex mix of lipophilic toxins ever found in mussels samples from Southern Chile.     

Industrially Relevant Canning Trials with a Sterilisation Time–Temperature Integrator

Food sterilisation, i.e. heating to above 120 °C for several min, still remains a primary method of food preservation. Current time–temperature integrators (TTIs), used to assess the thermal impact, only work at pasteurisation temperatures (below 100 °C). The aim of this work was to develop and validate a sterilisation TTI using an α-amylase from the hyperthermophile Pyrococcus furiosus. Previous experiments have found that this α-amylase has a z value similar to that of Clostridium botulinum spores and the enzyme is sufficiently heat resistant to show a measurable residual activity after 30 min at 121 °C. Analysis found a D value of 24 min and a slightly variable z value of around 11 °C. The decrease in enzymatic activity from thermal processing was found after a F_121°C process of 5 min to be 67 %, and for a F_121°C of 30 min it was 12 %, thus making P. furiosus α-amylase a strong and relevant candidate for a sterilisation TTI. Trials with canned water and mango chutney as test materials have confirmed the potential application of the enzyme within an industrially relevant setting of a model reel and spiral retort. The thermal data showed that at low or marginal F_121°C values, of less than 15 min, the correlation between the TTI and thermocouple data was closely comparable (R ²?=?0.96). The applicability of these TTIs over the range of industrial processes is also shown.     

Changes of heat-treated soymilks in bioactive compounds and their antioxidant activities under in vitro gastrointestinal digestion

To investigate the influence of in vitro gastrointestinal digestion on contents of bioactive compounds and antioxidant activities of heat-treated soymilks, changes of total phenolics content (TPC), total flavonoids content (TFC), content and profile of isoflavones, oxygen radical absorbance capacity (ORAC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and ferric-reducing antioxidant power (FRAP) were assayed after different heat treatments (at 95 °C for 20, 40, 60 min; 121 °C for 3, 6, 9 min and 143 °C for 20, 40, 60 s respectively) and gastrointestinal digestion. Results showed that digestion significantly influenced the contents of bioactive compounds and antioxidant activities of soymilks. Increases of DPPH radical scavenging activity, FRAP and ORAC of the heat-treated soymilks after gastric digestion were consistent with the increases of TPC (110.68–152.60 %) and TFC (4.48–31.10 %). In the dialysate fractions as the absorbable and utilized part, it was found that TPC, TFC, isoflavones and antioxidant activities (DPPH radical scavenging activity, FRAP and ORAC) were significantly decreased as compared with the gastric digestion fractions and duodenal fractions. Analysis showed that the bioaccessibility of TPC reached 107.17–125.14 %, TFC reached 34.63–67.19 % and total isoflavones reached 34.40–41.22 %, respectively, indicating the rich bioaccessible compounds in soymilk. Daidzein and its derivates were proven as the most bioaccessible isoflavones (about 36.99–44.14 %). Glucoside isoflavones showed the highest bioaccessibilities followed by malonylglucosides, acetylglucosides and aglycones. Overall, the soymilks treated at 95 °C and 60 min and 121 °C and 9 min had higher bioactive compounds contents, antioxidant activities and bioaccessibilities in the dialysate fractions.     

Antimicrobial Activity of Peptide P34 During Thermal Processing

In the present work, the influence of pH and sodium chloride concentration on thermal stability of antimicrobial peptide P34 was evaluated under different time–temperature conditions by a 2² factorial design experiment. At sterilization conditions (121 °C for 20 min), maximum retention (36%) was obtained at pH between 5.5 and 8.5 and sodium chloride concentration between 0.4 and 0.75 mol/l. For boiling conditions (100 °C for 20 min), antimicrobial activity was about 100% combining pH between 6.0 and 8.0 and salt concentration in the range of 0.65 to 1 mol/l. At low temperature pasteurization conditions (30 min at 65 °C), antimicrobial activity was not affected within the pH range from 5.0 to 8.0. For the three time–temperature conditions tested, the antimicrobial activity was minimal at more acidic or alkaline pH. Sodium chloride concentration of 0.65 mol/l increased thermostability of the peptide P34. Combination of sodium chloride and slight alkaline pH may increase the stability of peptide P34, which is essential to the proper utilization of bacteriocins in food industry.     

桑沟湾养殖牡蛎中贝类毒素监测及预警

利用高效液相色谱-串联质谱(high performance liquid chromatography-tandem mass spectrometry,HPLCMS/MS)方法监测养殖牡蛎中5种腹泻性贝类毒素与6种麻痹性贝类毒素含量变化及分布特征,分别利用HP20大孔型吸附树脂与SP700吸附树脂作为富集树脂,富集养殖海域海水中5种腹泻性贝类毒素与6种麻痹性贝类毒素,利用HPLC-MS/MS检测方法分析其中毒素含量,同步监测了养殖海域内牡蛎与海水中毒素含量,探究了两者之间的关系,建立了牡蛎内贝类毒素含量随海水内贝类毒素含量之间的变化规律。结果显示:在该海域内一共监测到OA、DTX-1、GYM、PTX-2 4种腹泻性贝类毒素与STX、dc STX两种麻痹性贝类毒素;在整个监控期内,海水中所监测贝类毒素随时间变化呈现先增长,达到峰值后逐渐降低趋势。牡蛎中毒素含量与海水中毒素含量呈正相关关系,即牡蛎内毒素的增长随海水内毒素的增长而增长,但牡蛎内毒素含量的峰值出现时间在海水中毒素含量出现峰值之后,延后时间为14 d。根据固相吸附毒素跟踪技术原理,可以提前14 d预警牡蛎内毒素含量。     

LC-MS/MS method for the determination of tetrodotoxin (TTX) on a triple quadruple mass spectrometer

Tetrodotoxin (TTX), often referred to as the ‘puffer fish’ poison, is a marine toxin and it has been identified as the agent responsible for many food poisoning incidents around the world. It is a neurotoxin that blocks voltage-gated sodium channels, resulting in respiratory paralysis and even death in severe cases. It is known to occur in many different species of fish and other organisms. The toxin is mainly found in the Southeast Asia region. Worryingly, TTX is starting to appear in European waters. It is suspected that this is a consequence of Lessepsian migration, also known as the Erythrean invasion. Therefore, straightforward and reliable extraction and analytical methods are now urgently required to monitor seafood of European origin for TTX. This paper provides a versatile, dependable and robust method for the analysis of TTX in puffer fish and trumpet shellfish using LC-MS/MS. A three-stage approach was implemented involving: (1) the screening of samples using fast multiple reaction monitoring (MRM) mass spectral analysis to identify quickly positive samples on a triple quadrupole mass spectrometer (QqQMS/MS), the API 3000; (2) a Fourier-transform (FT)-MS full-scan analysis of positive samples to collect qualitative data; and (3) a method with a longer chromatography run to identify and quantitate the positive samples using the QqQMS. The quantitative LC-QqQMS method delivered excellent linearity for solvent-based standards (0.01–7.5 µg ml^–1; R² ≥ 0.9968) as well as for matrix-matched standards (0.05–37.50 µg g^–1; R² ≥ 0.9869). Good inter-day repeatability was achieved for all the relevant analytes with %RSD values (n = 9) ranging from 1.11% to 4.97% over a concentration range of 0.01–7.5 µg ml^–1. A sample clean-up procedure for the puffer fish and trumpet shellfish was developed to ensure acceptable and reproducible recoveries to enable accurate and precise determination of TTX in a myriad of tissues types. Blank mackerel matrix was used for the TTX standard spiking studies in order to calculate the recoveries of the toxin during the extraction procedure. The recovery was 61.17% ± 5.42% for the extraction protocol. MS/MS studies were performed on a linear-trap quadruple-Orbitrap mass spectrometer (LTQ-Orbitrap) to obtain high-mass-accuracy data of the target analytes and their characteristic fragment ions in the puffer fish and trumpet shellfish samples. This facilitated identification of TTX and its associated analogues. These high-mass-accuracy studies facilitated the development of a rapid MRM-based quantitative method for TTX determination on the LC-QqQMS.     

Alternaria toxins in wheat from the Autonomous Province of Vojvodina,Serbia: a preliminary survey

Although Fusarium species remain a main source of mycotoxin contamination of wheat, in recent years, due to the evident climatic changes, other mycotoxigenic fungi have been recognised as important wheat contaminants. Alternaria species, especially A. alternata, have been found as contaminants of wheat as well as wheat-based products. Under favourable conditions A. alternata very often produce alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA) and others Alternaria toxins. The aim of the present study was to examine the presence of three Alternaria toxins (AOH, AME and TeA) in wheat samples harvested during three years (2011–13). To this end, 92 samples were collected during wheat harvesting from different growing regions of the Autonomous Province of Vojvodina, which represents the most important wheat-growing area in Serbia. The presence of Alternaria toxins was analysed by HPLC with electrospray ionisation triple quadrupole mass spectrometry (LC-ESI-MS/MS). Among all the analysed wheat samples, 63 (68.5%) were contaminated with TeA, 11 (12.0%) with AOH and 6 (6.5%) with AME. Furthermore, the maximum and mean toxin concentrations were 2676 and 92.4 µg kg^?1, 48.9 and 18.6 µg kg^?1, and 70.2 and 39.0 µg kg^?1 for TeA, AOH and AME, respectively. Co-occurrence of three Alternaria toxins in wheat samples was detected in six samples; a combination of two toxins was found in two samples; and 64 samples contained one toxin. The results showed that among 92 analysed wheat samples, only 20 (21.7%) samples were without Alternaria toxins. The presence of Alternaria toxins was also investigated in terms of weather conditions recorded during the period of investigation, as well as with the sampling region. This study represents the first preliminary report of the natural occurrence of Alternaria toxins in wheat (Triticum aestivum) from Serbia.     

Migration of lead and arsenic from food contact paper into a food simulant and assessment of their consumer exposure safety

ABSTRACT

Paper is one of the most commonly used food packaging materials. During the production of packaging paper, it is possible for trace amounts of heavy metals to be incorporated as contaminants. These could migrate into food when packaging paper (food contact paper) is used for cooking, storing and eating. The aim of this study was to determine the migration of lead (Pb) and arsenic (As) from food contact paper into a food simulant and then to assess human safety through the estimated daily intake (EDI) with consumption factor. Migration tests were conducted for 310 samples using 4% acetic acid as a food simulant at 25°C for 10 min and at 95°C for 30 min. Concentrations of Pb and As in a food simulant were quantified by inductively coupled plasma mass spectrometry. LODs for Pb and As were 0.002 and 0.005 µg L^?1, respectively. The migration of Pb from food contact paper ranged from not detected (ND) to 17.5 μg L^?1 at 25°C for 10 min and from 0.10 to 25.6 μg L^?1 at 95°C for 30 min while As ranged from ND to 0.44 μg L^?1 at 25°C for 10 min and from ND to 0.87 μg L^?1 at 95°C for 30 min. The migration of Pb and As determined in this study confirm that the human exposure was within safe levels based on the EDI of food contact paper compared with the provisional tolerable weekly intake for Pb of 25 μg kg^?1 bw and for As of 15 μg kg^?1 bw.     

Comparison of HPLC and Mouse Bioassay Methods for Determining PSP Toxins in Shellfish

Paralytic shellfish poisoning (PSP) toxins in shellfish were analyzed and quantified using both a high pressure liquid chromatographic (HPLC) technique and the standard AOAC mouse bioassay. Good correlation between the two assays was obtained at or below the limit of 80 μg toxin/100g, while at higher levels of toxicity, the HPLC technique tended to underestimate total toxicity slightly. Advantages and disadvantages of both techniques are discussed.     

Analysis of paralytic shellfish toxins and their metabolites in shellfish from the North Yellow Sea of China

Samples of toxic scallop (Patinopecten yessoensis) and clam (Saxidomus purpuratus) collected on the northern coast of China from 2008 to 2009 were analysed. High-performance liquid chromatography with post-column oxidation and fluorescence detection was used to determine the profile of the main paralytic shellfish poisoning (PSP) toxins in these samples and their total toxicity. Hydrophilic interaction liquid ion chromatography with mass spectrometric detection confirmed the toxin profile and detected several metabolites in the shellfish. Results show that C1/2 toxins were the most dominant toxins in the scallop and clam samples. However, GTX1/4 and GTX2/3 were also present. M1 was the predominant metabolite in all the samples, but M3 and M5 were also identified, along with three previously unreported presumed metabolites, M6, M8 and M10. The results indicate that the biotransformation of toxins was species specific. It was concluded that the reductive enzyme in clams is more active than in scallops and that an enzyme in scallops is more apt to catalyse hydrolysis of both the sulfonate moiety at the N-sulfocabamoyl of C toxins and the 11-hydroxysulfate of C and GTX toxins to produce metabolites. This is the first report of new metabolites of PSP toxins in scallops and clams collected in China.     

Chemical Composition, Antioxidant Capacity, and Sensory Quality of Pomegranate (Punica granatum L.) Arils and Rind as Affected by Drying Method

The objective of this study was to evaluate the application of: (1) freeze drying, (2) convective drying (50, 60, or 70 °C), (3) vacuum–microwave drying (240, 360, or 480 W), and (4) a combined method of convective pre-drying and vacuum–microwave finish drying in the processing of pomegranate arils and rind. The quality parameters under study included sugars and organic acids, punicalagins and ellagic acid, total polyphenols, total antioxidant activity, and sensory quality. In general, drying led to a reduction in all studied parameters; however, the behavior of arils and rind was different. Vacuum–microwave drying at 240 or 360 W was the best drying treatment for arils, while rind required freeze drying or soft conditions of convective drying (50 °C). Further research is needed to obtain proper results with convective pre-drying and vacuum–microwave finish drying of arils and rind. With proper selection of the drying protocol, high-quality dried arils will be available for consumers; these arils will be characterized by high contents of fructose (25 g 100 g^?1), phytic acid (2.2 g 100 g^?1), punicalagins (0.57 mg g^?1), total polyphenols (1.6 mg eq gallic acid g^?1), high antioxidant capacity (0.6 mg eq Trolox g^?1), and high intensities of garnet color, sweetness, sourness, and fresh pomegranate aroma. Besides, dried rind with very high contents of active compounds (123 mg g^?1 of punicalagins and 108 mg eq gallic acid g^?1) and high antioxidant capacity (26 mg eq Trolox g^?1) will be also available as functional material.     

Production and physicochemical properties of resistant starch from hydrolysed wrinkled pea starch

The content and physicochemical properties of resistant starches (RS) from wrinkled pea starch obtained by different molecular mass reduction processes were evaluated. Native and gelatinised starches were submitted to acid hydrolysis (2 m HCl for 2.5 h) or enzymic hydrolysis (pullulanase, 40 U g^?1 for 10 h), followed by hydrothermal treatment (autoclaving at 121 °C for 30 min), refrigeration (4 °C for 24 h) and lyophilisation. Native starch showed RS and total dietary fibre contents of 39.8% and 14.3%, respectively, while processed ones showed values from 38.5% to 54.6% and from 22.9% to 37.1%, respectively. From these, the highest contents were among acid‐modified starches. Processed starches showed endotherms between 144 and 166 °C, owing to the amylose retrogradation. Native and processed starches showed low viscosity, which is inversely proportional to the RS concentration in samples. The heat treatment promoted an increase in the water absorption index. The pea starch is a good source for obtaining resistant starch by acid hydrolysis.     

Rapid Detection of <Emphasis Type="Italic">Vibrio parahaemolyticus</Emphasis> in Shellfish by Real-Time Recombinase Polymerase Amplification

Vibrio parahaemolyticus (V. parahaemolyticus) is a zoonotic pathogen generally found in seafood. To detect the foodborne pathogen rapidly and accurately for food safety measures, we developed a real-time recombinase polymerase amplification (RPA) method. An evaluation of the specificity and sensitivity of the method is discussed here. A set of primers and probe was specially designed to target the tlh gene, which is usually regarded as a marker of total V. parahaemolyticus strains. During the reaction, target DNA was amplified and tagged with specific fluorophore within 10 min and at an incubation temperature of 40 °C. In addition to fast amplification and low temperature, the fluorescence signal was synchronized with the amplification of products for the generation of real-time data. The detection limit of this assay was 0.4 pg/μL of DNA, which is comparable to assays that use the bacterial culture as template, 4?×?10³ cfu/mL. The real-time RPA method had a stable performance when testing the spiking shellfish samples at the same level of contamination by the pathogen in different kinds of shellfish. Thus, the real-time RPA method shows great potential for on-site detection of V. parahaemolyticus, especially in low-resource settings.     

Impact of γ‐irradiation and thermal processing on the antigenicity of almond,cashew nut and walnut proteins

Whole unprocessed almonds, cashew nuts and walnuts were each subjected to γ‐irradiation (1, 5, 10 and 25 kGy) followed by heat processing including autoclaving (121 °C, 15 psi for 15 and 30 min), dry roasting (138 and 160 °C for 30 min each, 168 and 177 °C for 12 min each), blanching (100 °C for 5 and 10 min), oil roasting (191 °C, 1 min) and microwave heating (500 W for 1 and 3 min). Rabbit polyclonal antibodies were raised against each major protein isolated from defatted, but not subjected to γ‐irradiation and/or any thermal processing, almond, cashew nut and walnut flours. Immunoreactivity of almond, cashew nut and walnut proteins soluble in borate saline buffer, normalised to 1 mg protein ml^?1 for all samples, was determined by inhibition enzyme‐linked immunosorbent assay (ELISA) and Western blotting. ELISAs and Western blotting experiments indicated that almond, cashew nut and walnut proteins exposed to γ‐irradiation alone or followed by various thermal treatments remained antigenically stable. Copyright © 2004 Society of Chemical Industry     

Toxin composition and toxicity dynamics of marine gastropod Nassarius spp. collected from Lianyungang,China

Consumption of nassariid gastropods often leads to poisoning incidents in some coastal provinces in China. To elucidate the pattern of toxicity dynamics and origin of toxins, samples of gastropod Nassarius spp. were collected from late May to early August 2007 from Lianyungang, Jiangsu province, where the poisoning incidents have been frequently reported. Toxicity was first screened with the mouse bioassay method, and tetrodotoxin and its analogues (TTXs) were analysed with high-performance liquid chromatography coupled with an ion-trap mass spectrometer (HPLC-MS ⁿ ). The toxicity of nassariid N. semiplicatus showed an ‘M’-shaped pattern of fluctuation during the sampling season. Two peaks of toxicity appeared in late May and late July. The maximum toxicity was recorded on 24 May, with the value of 846 mouse unit (MU) g^?1 of tissue (wet weight). TTX and its analogues trideoxyTTX, 4-epiTTX, anhydroTTX and oxoTTX were detected in the nassariid samples. TrideoxyTTX but not TTX was the major toxin in all the samples. No paralytic shellfish poison (PSP) was detected in the sample with the maximum toxicity by HPLC-FLD analysis. Variation of TTX content in the tissue of nassariid gastropods correlates well with the dynamics of toxicity. It is suggested that TTXs are the major toxins corresponding to the toxicity of the nassariids, and May and July are the high-risk seasons for consumption of nassariids, which is critical for the management of poisoning incidents.     

Supercritical Carbon Dioxide Processing of Dry Cured Ham Spiked with Listeria monocytogenes: Inactivation Kinetics, Color, and Sensory Evaluations

The feasibility of supercritical carbon dioxide (SC-CO₂) treatment to inactivate Listeria monocytogenes inoculated on the surface of dry cured ham was investigated. A multibatch apparatus was used. Inactivation kinetics were determined at 8 and 12 MPa, as a function of temperatures (35–50 °C), treatment times (5–60 min), and initial microbial loads (10³–10⁷ colony-forming units [CFU]/g). Color changes of the sample were determined by measuring the reflectance spectra and L*, a*, and b* parameters. A new spectroscopic technique was developed for this. Sensory quality of the product before and after the SC-CO₂ treatment was evaluated by a sensory panel. Treatment at 50 °C, 12 MPa for 15 min resulted in total inactivation of L. monocytogenes with an initial microbial load of 10⁷ CFU/g. Less severe conditions, e.g., 45 °C, 12 MPa, 5 min, were sufficient to reach total inactivation if the initial microbial load was 10³ CFU/g. The process slightly influenced the color and sensory attributes of the sample. The results demonstrated the efficiency and the potential of SC-CO₂ as a technology for the pasteurization of the surface of foods, in particular ham-type meat products.