Prevalence,enumeration, and characterization of Salmonella isolated from aquatic food products from retail markets in China

This study aimed to estimate the extent and level of Salmonella contamination of aquatic food products in China, and to determine serotype, virulotype, and the antimicrobial resistance profiles of recovered Salmonella isolates. Out of 554 samples collected from July 2011 to May 2014, 86 (15.5%) tested positive for Salmonella. The highest contamination rate occurred in oysters (23.1%, 6/26), followed by freshwater fish (18.6%, 43/231), shrimp (13.0%, 13/100), and saltwater fish (12.2%, 24/197). The contamination levels generally corresponded to a most probable number (MPN)/g of 0.3–10, although one sample exceeded 110 MPN/g. Among the 103 isolates, S. Typhimurium, S. Wandsworth, S. Thompson, and S. Derby were the most prevalent serovars. Sixty-eight isolates (66.0%) were resistant to at least one antimicrobial, and 35 (34.0%) were resistant to more than three. High rates of resistance were observed for tetracycline (35.9%), ampicillin (28.2%), nalidixic acid (26.2%), trimethoprim-sulfamethoxazole (25.2%), chloramphenicol (20.4%) and streptomycin (18.4%). Of note, S. Thompson isolates exhibited resistance to multiple extended-spectrum cephalosporins, ciprofloxacin, and other antimicrobials. PCR analysis of 15 virulence genes showed that ssaQ, mgtC, siiD, sopB, and bcfC were present in all 103 isolates, whereas the remaining loci were variably distributed. S. Typhimurium, S. Enteritidis, and S. Weltevreden isolates exhibited a wider range of pathogenicity determinants compared with the other strains. Our study provides a comprehensive surveillance on prevalence of Salmonella in aquatic food products from China and indicates its potential risk to public health. These data are valuable for epidemiological studies, risk management, and public health strategies.     

Efficiency of acetic acid and formic acid as a catalyst in catalytical and mechanocatalytical pretreatment of barley straw

In this study, the potential of organic acids (formic acid, acetic acid) in a catalytical and mechanocatalytic conversion of lignocellulosic barley straw to valuable sugars is explored using sulfuric acid as a reference. Acid-catalyzed hydrolysis has been carried out with acid-impregnated samples as well as unmodified barley straw. In the mechanocatalytical approach, pretreatment consists of impregnation with the acid catalyst and mechanical treatment by ball milling following chemical hydrolysis. Straw samples and residues were analyzed by Fourier transform infrared spectrometry (FT-IR) whereas hydrolysate analysis was based on total reducing sugar (TRS) determination following the DNS method and capillary electrophoresis (CE) analysis. The results indicated that acetic acid and formic acid are rather mild acids yielding low TRS levels compared to the reference acid. Mechanocatalytical pretreatment slightly increased TRS yields, but not significantly. Strikingly, sulfuric acid showed an efficient conversion efficiency yielding almost 45% of TRS. Furthermore, this study provided evidence for the acetylation of straw components when acetic acid was used as catalyst. Alkali hydrolysis induced the de-esterification, but revealed no significant increase of TRS yields.     

Aflatoxin B1 degradation by culture and lysate of a Pontibacter specie

The presence of aflatoxin B₁ (AFB₁) along the food chain poses a significant threat, thus propelling the need for an effective approach to control it. This study was therefore, aimed at investigating AFB₁ degradation of liquid cultures and lysates of an isolated Pontibacter sp. (VGF1). Liquid cultures, lysed bacterial cells in the absence (uninhibited lysates) and presence of protease inhibitors (protease inhibited lysates) were respectively incubated with AFB₁ for 3, 6, 12, 24 and 48 h. AFB₁ degradation was monitored during this period on high performance liquid chromatography (HPLC) and results obtained revealed that after 6 h of incubation, the protease inhibited (PI) lysates yielded a 65% AFB₁ degradation, whereas after 12 h, no residual AFB₁ was detected. Conversely, after 48 h of incubation, a significantly (p≤0.05) lower AFB₁ degradation of 50 and 36% by the liquid culture and uninhibited lysate, respectively, were noted. It was further confirmed that the degradation mechanism was enzymatic. Data from cytotoxicity studies against human lymphocytes further demonstrated that extracts of biotransformed AFB₁ were less toxic when compared to that of AFB₁. Findings from this study have demonstrated an alternative approach for the decontamination and biocontrol of AFB₁ in various agricultural commodities.     

A reliable and sensitive time-resolved fluorescent immunochromatographic assay (TRFICA) for ochratoxin A in agro-products

Immunochromatographic assays (ICAs) are considered as a suitable diagnostic tool for the detection of mycotoxins. Mycotoxins and especially, ochratoxin A are analytes with more demanding sensitivity requirements. To enhance the sensitivity of current immunochromatographic assays for ochratoxin A (OTA), a novel sensitive ICA was developed in this study. In the assay, microspheres enclosing fluorescent europium (III) [Eu(III)] nanoparticles (EuNPs) were used as a label for OTA monoclonal antibody (OTA-mAb) conjugation. Accordingly, assay was called time-resolved fluorescent immunochromatographic assay (TRFICA). The test strip was composed of three parts: a sample pad, nitrocellulose membrane and an absorbent pad. As for detection, a proper concentration of conjugated microspheres was pipetted into the microtube and sample extract was added to it. Then the strip was inserted into the tube and the fluid flow along the strip. The TRFICA results were obtained in 8 min and read by a portable TRFICA strip reader. The established method allows quantitative determination of OTA with limit of detection as low as 1.0 μg kg⁻¹ in the samples. For validation, spiked samples including wheat, maize, soybean and rice were respectively assayed by TRFICA and a standard high performance liquid chromatography equipped with a fluorescence detector (HPLC-FLD), and good agreement of results was obtained between two methods.     

超高效液相色谱-荧光法测定蜂蜜中百里香酚的残留量

建立了超高效液相色谱-荧光检测法测定蜂蜜中百里香的残留量的检验方法。用40%乙腈-水溶液提取蜂蜜中的百里香酚,经离心、上清液过滤后,以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,采用Accucore aQ色谱柱(150 mm×2.1 mm,2.6μm)分离,荧光检测器检测(激发波长274 nm,发射波长297 nm),外标法定量。结果表明,百里香酚在0.01～3.00μg/mL范围内线性关系良好,决定系数(R²)为0.9999,在蜂蜜基质中的平均加标回收率在87.4%～106.9%之间,变异系数在0.7%～7.3%之间,定量限为0.10 mg/kg。利用该方法测定了48份市售蜂蜜样品,仅有1份样品存在百里香酚残留。本研究建立的方法简单、迅速、可靠,适用于蜂蜜中百里香酚残留的批量筛查和定量测定。     

Determination of 15 β-lactam antibiotics in pork muscle by matrix solid-phase dispersion extraction (MSPD) and ultra-high pressure liquid chromatography tandem mass spectrometry

An UPLC–MS/MS method based on matrix solid-phase dispersion (MSPD) was developed for simultaneous determination of 15 β-lactams in pork muscle. Muscle tissue was blended with Oasis HLB adsorbent. A column made from the sorbent/muscle tissue matrix was washed with hexane, after which β-lactams was eluted with acetonitrile/water (50:50, v/v) both containing 0.1% formic acid. The final extract was evaporated, and reconstituted into acetonitrile/water (10:90, v/v) for the analysis of UPLC-MS/MS. The correlation coefficient (r²) with each matrix-matched calibration curve is higher than 0.99. Limits of detection (LODs) and limits of quantification (LOQs) of β-lactams were ranged from 0.02 to 0.63 μg kg⁻¹ and 0.07–0.97 μg kg⁻¹, respectively. At the fortified levels, mean recoveries of all compounds were ranged from 92 to 111%, and the RSDs were lower than 12%. The method was accurate and reproducible, being successfully applied to the monitoring of 15 β-lactams in pork muscles obtained from the Chinese market.     

Nitric oxide (NO) production in mammalian non-tumorigenic epithelial cells of the small intestine and macrophages induced by individual strains of lactobacilli and bifidobacteria

Nitric oxide (NO) affects multiple gastrointestinal functions, including mucosal inflammation and antimicrobial activity. The aim of this study was to screen the ability of probiotic bacteria to stimulate NO production in porcine intestinal epithelial cells and macrophages in the presence and absence of interferon gamma (INF-γ). Production of NO in intestinal epithelium was stimulated by individual strains of lactobacilli without INF-γ priming. While none of the tested bifidobacteria were capable of inducing NO production, most constitutively secreted NO. Most tested strains induced a significant increase in NO production compared with the control cells in the macrophage cell line 3D4/21. Results support the protective role of the individual strains of the genera Lactobacillus and Bifidobacterium and may lead to new approaches for manipulating and regulating immune responses at the mucosal surfaces of the gastrointestinal tract.     

磁固相萃取在食品分析中的研究进展

磁固相萃取(Magnetic solid phase extraction,MSPE)是使用磁性吸附剂富集待测物,在磁场作用下分离待测物的样品前处理方法。相比于传统的样品前处理方法,MSPE具有吸附快速、效率高、易分离等优点。MSPE磁性吸附剂由磁性纳米颗粒和其表面的非磁性功能材料修饰组成。本文主要综述碳化合物、表面活性剂、离子液体、金属-有机骨架化合物、氧化物、高分子、分子印迹聚合物这几类材料表面修饰的MSPE磁性吸附剂的研究进展。选择不同表面修饰材料的MSPE技术可以对不同的食品添加剂、农药残留、抗生素等化合物有理想的吸附效果,有广泛的应用前景。     

高效液相色谱-串联质谱法快速检测银耳中的米酵菌酸

目的建立高效液相色谱-串联质谱法(highperformanceliquidchromatography-tandemmass spectrometry,HPLC-MS/MS)快速检测银耳中的米酵菌酸的方法。方法样品以甲醇提取后,超声振荡,离心分层,上清液用甲醇定容。使用空白基质液配置标准曲线对结果进行校正。在优化后的色谱及质谱条件下,采用(electronsprayionization,ESI)负离子模式进行电离,并通过多反应监测(multi-selectedreactionmonitoring,MRM)模式对目标化合物的定量离子和定性离子进行测定。结果米酵菌酸在0.50～50.00μg/L范围内线性良好,相关系数r为0.9996,3个浓度添加水平的平均回收率范围在80.6%～95.6%之间,相对标准偏差为4.98%～7.21%。方法检出限(limit of detection, LOD)为5.0μg/kg,定量限(limit of quantification, LOQ)为15.0μg/kg。结论该方法操作简便快速,回收率高、灵敏度高、精密度好,适合测定银耳基质中的米酵菌酸。     

基于MALDI-TOF MS技术快速检测产呕吐毒素蜡样芽胞杆菌

该研究探讨了基质辅助激光解吸电离飞行时间质谱仪快速鉴别产呕吐毒素蜡样芽胞杆菌的方法。通过对标准品进行分析,重新对特征峰进行定位并测定其灵敏度,并用正交试验分析不同培养条件下检验结果的差异,用优化后的培养条件对49株野生蜡样芽胞杆菌及3株标准菌株进行特异性检验。研究表明,MALDI-TOF MS可检测到产呕吐毒素蜡样芽胞杆菌中呕吐毒素相应m/z值为1 175的[M+Na]+和m/z值为1 191的[M+K]+加合物特征峰,具有较高的灵敏度（0.01 μg/mL）,经极差分析显示,选用MYP培养基30 ℃培养12 h后的菌落能获得最稳定、响应值高（>104）的检验结果;方法应用验证表明,49株野生菌株中2株含ces基因的蜡样芽胞杆菌均检出,其余未含有ces基因的菌株均未检出。该研究建立的MALDI-TOF MS检测方法能直接快速准确检出产呕吐毒素蜡样芽胞杆菌,该方法检测特异性强（100%）,灵敏度高（0.01 μg/mL）,对于食品安全事故快速精准分析研判有重要的意义。