Homing at an extragenic locus mediated by VDE (PI-SceI) in Saccharomyces cerevisiae

PI-SceI (VDE), a homing endonuclease with protein splicing activity, is a genomic parasite in the VMA1 gene of Saccharomyces cerevisiae. In a heterozygous diploid of the VDE-less VMA1 allele and a VDE-containing VMA1 allele, VDE specifically cleaves its recognition sequence (VRS) in the VDE-less VMA1 allele at meiosis, followed by 'homing', i.e. a conversion to a VDE-containing allele. We found that upon VDE expression, homing of a marker gene at an extragenic locus occurs only when a 45 bp element containing the VRS is inserted at its allelic site, while mutants of VDE with no endonuclease activity lack authentic extragenic homing activity. Thus, both the VRS and VDE are required for homing. Insertion of the VRS in a homozygous diploid significantly lowered the spore germination ability, indicating that a template for gene repair at its allelic locus is essential for efficient homing and survival of yeast cells.     

动物源性食品中抗生素残留、耐药细菌的分布及耐药基因的水平传播

目的：调查动物源性食品中抗生素的残留、耐药细菌的分布及抗生素对耐药基因水平转移的影响。方法：利用盐析辅助液液萃取/高效液相色谱/串联质谱法对市售动物源性食品中18种抗生素的含量进行检测;对样品中的需氧菌进行分离鉴定,利用PCR技术调查耐药基因的分布,通过接合转移试验调查抗生素胁迫对耐药基因水平传播的影响。结果：50份动物源性食品中检出环丙沙星、恩诺沙星、土霉素、金霉素、磺胺嘧啶和磺胺二甲嘧啶。162株分离株中磺胺甲基异恶唑耐药基因sul1的检出率最高。在环丙沙星胁迫下,三株供体菌中耐药基因qnrS的接合转移率均高于对照组。结论：动物源性食品中喹诺酮类、磺胺类和四环素类抗生素均有不同程度的残留。亚抑菌浓度抗生素促进耐药基因的接合转移。     

The effect of glyphosate on digestion and horizontal gene transfer during in vitro ruminal fermentation of genetically modified canola

BACKGROUND: The impact of glyphosate on ruminal fermentation, selective pressure on ruminal bacteria and horizontal transfer of the gene encoding 5‐enolpyruvylshikimate‐3‐phosphate synthase (epsps) to ruminal bacteria was studied using batch culture with glyphosate‐tolerant (Roundup Ready^®) canola meal as substrate. RESULTS: A glyphosate concentration × time interaction (P < 0.05) occurred when glyphosate (0–100 mmol L^?1) was included in in vitro ruminal incubations with a diet containing 150 g kg^?1 Roundup Ready^® canola meal (Experiment 1). Glyphosate at 50 and 100 mmol L^?1 inhibited fermentation. In Experiment 2, epsps fragments were detected in plant debris for up to 16 h of incubation using primer sets that amplified three fragments (62, 108 and 300 bp) of DNA spanning the transgenic construct. Persistence was affected by fragment size but not by glyphosate concentration (0, 10 or 60 mmol L^?1). Extensive polymerase chain reaction assays provided no evidence of acquisition of epsps by feed‐ or fluid‐associated bacteria during fermentation. A glyphosate concentration × time interaction (P < 0.05) was observed for all fermentation parameters measured, and glyphosate caused a general inhibition of fermentation. CONCLUSION: The presence of glyphosate did not increase selective pressure for gene transfer of DNA encoding glyphosate resistance from Roundup Ready^® canola meal to ruminal bacteria. Copyright © 2007 Society of Chemical Industry     

Characterization and horizontal transfer of class 1 integrons in Salmonella strains isolated from food products of animal origin

A total of 81 Salmonella isolates from retail meats and seafood in Hebei province, China, were assayed for the presence of and horizontal transfer of class 1 integrons. By the PCR screening for the integrons, class 1 integron was detected from strains in serotypes of Derby, Indiana, London and Choleraesuis, which were isolated from pork, chicken or seafood; however, two isolates contained the empty integron that lacked the resistance cassette, a potential hotspot for development of the multidrug resistance. In contrast, two other isolates had the antibiotic resistance gene cassettes within the class 1 integron, which were dfrA1-aadA1 and aadB-cmlA, respectively. The conjugation experiments demonstrated the plasmid-mediated transfer of the class 1 integrons. Furthermore, each of the integrons was transmitted to Streptococcus mutans via natural gene transformation. These findings suggest the possible transfer of class 1 integrons from foodborne pathogens to human residential bacteria via horizontal gene transfer.     

Determination of aflatoxins in eggs,milk, meat and meat products using HPLC fluorescent and UV detectors

Raw and pasteurised sheep’s, cow’s and goat’s milk, eggs, and beef samples from different local markets in Jordan were collected during a period of 5 months (January through May 2007) and examined for aflatoxins B1(AFB1), B2(AFB2), G1(AFG1), G2(AFG2), M1(AFM1) and M2(AFM2). The samples were analysed with high performance liquid chromatography (HPLC) using UV and Fluorescent detectors. The analysed samples of milk collected in January were found to contain 0.56 μg L⁻¹ AFM1 and 0.1 μg L⁻¹ AFM2 whilst, the concentration of AFM1 and AFM2 was < 0.05 μg L⁻¹ for milk samples collected between March and May. The AFB1, AFB2, AFG1 and AFG2 contents in the analysed food products ranged from 1.10 to 8.32 μg L⁻¹ and 0.15 to 6.36 μg L⁻¹ in imported and fresh meat samples collected during March, respectively. The mean recovery for the HPLC method was 92% to 109% and the quantification levels were 50 ng L⁻¹ for AFM1 and AFM2. The AFM1 was found in 10% of the tested samples with concentrations between 0.08 and 1.1 μg kg⁻¹ and AFM2 was only found in 1.82% of the tested samples with a level of 0.1 μg kg⁻¹. The AFM1 levels in the examined foods were higher than the maximum level of AFM1 in liquid milk set by the European Community and Codex Alimentarius of 50 ng L⁻¹.     

计算流体力学在食品传热传质过程中的应用

对计算流体力学(CFD)技术进行了简单介绍，并对采用CFD技术模拟食品加工过程中的传热传质过程的研究现状进行了综述，探讨了采用CFD对这些传热传质过程进行数值模拟的优势。CFD模拟的结果详细、直观，模拟得到的流体流动和各种热力学参数的分布信息，可以为装置的设计、传热过程的优化提供参考。     

Natural co-occurrence of aflatoxin B1, fumonisin B1 and ochratoxin A in barley and corn foods from Korea

A survey for aflatoxin B₁ (AFB₁), fumonisin B₁ and ochratoxin A (OTA) was conducted on 127 samples that included 30 food-grade barley, 32 barley foods, 18 food-grade corn and 47 corn foods, randomly collected during 1998-99 in Seoul, Korea. The presence of mycotoxins was analysed by direct competitive enzyme-linked immunosorbent assay (ELISA), and most of the positive samples from ELISA were confirmed using high-performance liquid chromatography (HPLC). Recoveries of AFB₁ and OTA spiked at 10 ng g -1 and FB₁ spiked at 50 ng g^-1 were 106, 87 and 105% by ELISA, whereas those by HPLC were 80, 79 and 84%, respectively. Detection limits by ELISA for AFB₁, FB₁ and OTA were 1, 5 and 5 ng g^-1, and those by HPLC were 0.6, 35 and 1 ng g^-1. Naturally occurring AFB₁, FB₁ and OTA were found in 4/32 (12%), 2/32(6%) and 4/32 (12%) samples of barley foods with an average of 26, 16 and 9 ng g^-1, respectively. AFB₁ and FB₁ in corn foods were detected in 4/47 (8%) and 9/47 (19%) samples with the average being 20 and 74 ng g^-1, while no OTA was found in any corn foods samples. No AFB₁, FB₁ or OTA was detected in any of food-grade barley and corn samples. This is the first report on the natural co-occurrence of AFB₁ and FB₁ in barley and corn foods as well as on surveillance of OTA in Korea.     

Occurrence of dioxins (PCDDs,PCDFs) and polychlorinated biphenyls (PCBs) in wild,farmed and processed fish,and shellfish

Forty-eight composite samples of the most commonly consumed fish and shellfish species were prepared from up to 60 individual subsamples of each species and analysed for chlorinated dioxins (PCDD/Fs) and polychlorinated biphenyls (PCBs). These included 24 species of fresh wild fish, seven of farmed fish, seven of fresh shellfish, and ten processed fish and shellfish products. The ISO 17025-accredited analytical methodology used is consistent with the requirements given in European Commission Directive 2002/69/EC. Concentrations ranged from 0.03 ng kg^–1 PCDD/F plus PCB World Health Organization-toxic equivalent quantity (WHO-TEQ) for a sample of surimi, to approximately 6 ng kg^–1 for wild pilchards/sardines. The corresponding range for the ΣICES-6 PCBs was 0.04 μg kg^–1 to approximately 47 μg kg^–1. None of the samples showed concentrations above the European Union maximum permitted limits. Averaged PCDD/F and PCB concentrations for the two groups of farmed and wild fish show that there is little difference between the two categories, although individual species may show variations depending on factors such as the sampling location.     

Occurrence of patulin and its dietary intake through pear,peach, and apricot juices in Italy

Pb, Cd and Hg in muscles of flathead mullet (Mugil cephalus), collected from Bocca di Magra, La Spezia (Ligurian Sea, Mediterranean Sea, Italy), were determined using graphite furnace atomic absorption spectrometry after microwave digestion for Pb and Cd and direct mercury analyser for Hg. Average Pb concentrations varied in the range 0.20–0.24 mg/kg, whereas Cd and Hg levels were negligible. None of the tested 200 samples exceeded the European regulatory limits as set by EC 1881/2006 and 420/2011. Metal concentrations in fish muscles were assessed for human consumption according to provisional tolerable weekly intake. The estimated values of Pb, Cd and Hg in M. cephalus’s edible parts in this study were below the values established by the Joint WHO/FAO Expert Committee on Food Additives. Therefore, it can be concluded that there is no health problem in human consumption.     

Occurrence and stability of inorganic and organic arsenic species in wines,rice wines and beers from Central European market

We investigated in total 80 wine samples of different types and seven grape juice and 23 beer samples purchased from markets in Central Europe in order to understand the arsenic (As) speciation and help assess the potential As toxicity via intake of alcoholic beverages. Generally, total As concentrations in most samples investigated were below the drinking water limit 10?µg?l^?1 published by the World Health Organization (WHO); ranging from 0.46 to 21.0?µg?l^?1 As in red and white wines and from 0.75 to 13.4?µg?l^?1 As in beers. In addition, concentrations of total As in rice wine and in rice beer were 0.63–6.07 and 3.69–8.23?µg?l^?1 As, respectively. The total As concentrations in ice wine ranged from 7.94 to 18.8?µg?l^?1 As, significantly higher than in white and red wine. Arsenite predominated as the As species in most of the wine samples, whereas arsenate was the dominant species in rice wine, beer and rice beer. Methyl As components were usually minor components in all wine and beer samples. Monomethylarsonic acid, dimethylarsinic acid and two additional unknown As species were frequently found in grape juice, late harvest and ice wine with higher sweetness. After air exposure, arsenite, arsenate, monomethylarsonic acid and dimethylarsinic acid were stable at 4°C for months, probably due to the acidic conditions of wine and beer samples. The presence of sulfite had little influence on As speciation in wine. Despite the predominance of more toxic arsenite and arsenate in wine and beer, the estimated weekly exposure to As (via consumption of beer, wine and rice wine) is low. The As intake per capita is 6.81?µg from beer, <1.93?µg from wine and 0.88?µg from rice wine, estimated using the median of total As concentration multiplied by the average consumption per capita of the corresponding beverage.     

Evolution of oxidative and structural characteristics of proteins,especially lipid transfer protein 1 (LTP1) in beer during forced-ageing

The evolution of oxidative and structural characteristics of proteins, especially lipid transfer protein 1 (LTP1), in beer during forced-ageing was examined. The oxidative characteristics of beer and proteins were evaluated by DPPH radical scavenging activity and ABTS radical cation scavenging activity. Results showed that the levels of proteins, thiols, LTP1 and antioxidant activity decreased gradually. This was accompanied by the degradation of macromolecular proteins in beer during forced-ageing. Results from circular dichroism (CD), Fourier-transform infrared spectroscopy (FTIR), surface hydrophobicity (S₀) and ζ-potential further indicated that the secondary and tertiary structure of LTP1 changed drastically during forced-ageing, with the reduction of the S₀, α-helix and β-sheet contents and the increase in negative ζ-potential and random coil. Thus, the proteins, especially LTP1, might play important roles in maintaining oxidative stability of beer.     

Occurrence of the vancomycin-resistant genes vanA, vanB, vanCl, vanC2 and vanC3 in Enterococcus strains isolated from poultry and pork

It is suspected that the use of avoparcin as a feeding antibiotic for the fat stock contributes to development of cross-resistance against vancomycin and teicoplanin. After isolating enterococci strains from poultry and pork meat by cultivation on citrate azide Tween carbonate agar (CATC) and screening the vancomycin resistance on Columbia colistin nalidixic acid agar (CNA, supplemented with 5% sheepblood and 5 mg vancomycin/l) the polymerase chain reaction (PCR) was used for the detection of the vancomycin resistance genes vanA (‘high level’), vanB (‘moderate high level’), vanC1, vanC2 and vanC3 (‘low level’). Out of 1643 E.-isolates from 115 poultry and 50 pork samples, 420 isolates could be identified as vancomycin resistant, 202 isolates of which carry the vanA, one isolate both the vanA and the vanC1, 38 isolates the vanC1, 14 isolates the vanC2, nine isolates both the vanC1 and the vanC3 gene and 156 isolates carry no gene. The vanB gene was not found in these isolates. Comparing vanA-positive food isolates with those from different human sources by means of the pulsed field gel electrophoresis (PFGE) it could clearly be demonstrated that they do not show homological fingerprints according to the source of origin. It is therefore unlikely that there is a close genetic relationship between isolates from animal foodstuff and humans.     

Impact of single nucleotide polymorphisms in leptin, leptin receptor, growth hormone receptor, and diacylglycerol acyltransferase (DGAT1) gene loci on milk production, feed, and body energy traits of UK dairy cows

The impact of 9 single nucleotide polymorphisms (SNP) in the leptin (LEP), leptin receptor (LEPR), growth hormone receptor (GHR), and diacylglycerol acyltransferase (DGAT1) gene loci on daily milk production, feed intake, and feed conversion, and weekly measures of live weight, BCS, and body energy traits was evaluated using genetic and phenotypic data on 571 Holstein cows raised at the Langhill Dairy Cattle Research Center in Scotland. Six SNP were typed on the LEP gene and 1 on each of the other 3 loci. Of the 6 LEP SNP, 3 were in very high linkage disequilibrium, meaning there is little gain in typing all of them in the future. Seven LEP haplotypes were identified by parsimony-based analyses. Random-regression allele-substitution models were used to assess the impact of each SNP allele or haplotype on the traits of interest. Diacylglycerol acyltransferase had a significant effect on milk yield, whereas GHR significantly affected feed intake, feed conversion, and body energy traits. There was also evidence of dominance in allelic effects on milk yield and BCS. The LEP haplotype CCGTTT (corresponding to leptin SNP C207T, C528T, A1457G, C963T, A252T, and C305T, respectively) significantly affected milk yield and feed and dry matter intake. Animals carrying this haplotype produced 3.13 kg more milk daily and consumed 4.64 kg more feed. Furthermore, they tended to preserve more energy than average. Such results may be used to facilitate genetic selection in animal breeding programs.     

Effect of 1-methylcyclopropene on shelf life,visual quality,antioxidant enzymes and health-promoting compounds in broccoli florets

The effect of 1-methylcyclopropene (1-MCP) on quality, antioxidant enzymes and glucosinolate contents in broccoli (Brassica oleracea var. italica) florets was investigated in the present study. Broccoli florets were treated with air (control) and 2.5 μl/l 1-MCP for 6 h at 20 °C, and were then stored at 20 °C for 5 days. 1-MCP treatment markedly extended shelf life, reduced postharvest deterioration, retarded chlorophyll degradation and inhibited the increase of malondialdehyde amount and activities of polyphenol oxidase and lipoxygenase in florets. The activities of superoxide dismutase, peroxidase and catalase in florets treated with 1-MCP were higher than those in control florets. 1-MCP treatment reduced the rate of decrease of total carotenoids, ascorbic acid and glucosinolates in florets when compared to those in the control. These results indicated that 1-MCP treatment could be a good candidate for extending shelf life, maintaining visual quality and reducing loss of health-promoting compounds, particularly glucosinolates in broccoli.     

不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较

本实验利用小鼠3T3-L1前脂肪细胞对绿茶多糖（green tea polysaccharides,GTP）、红茶多糖（black tea polysaccharides,BTP）和乌龙茶多糖（oolong tea polysaccharides,OTP）的减肥作用进行评价。使用气相色谱对GTP、BTP、OTP的单糖组成进行分析。采用噻唑蓝染色法测定3?种茶多糖（tea polysaccharides,TPs）对3T3-L1前脂肪细胞增殖活力的影响。使用流式细胞仪检测3?种TPs对3T3-L1前脂肪细胞细胞周期的影响。采用传统“鸡尾酒”法诱导3T3-L1细胞分化成脂后,测吸光度并计算其分化率。采用甘油磷酸氧化酶-过氧化物酶（glycerol phosphate oxidase-peroxidase,GPO-PAP）法测定细胞中甘油三酯（triglyceride,TG）含量的变化。采用实时荧光定量聚合酶链式反应（real-time polymerase chain reaction,RT-PCR）技术测定与脂质代谢相关基因的表达量。结果显示3?种TPs均能显著抑制3T3-L1前脂肪细胞的增殖与分化（P＜0.05）。添加100?μg/mL的TPs能使细胞分化率显著下降至（62.00±6.61）%（GTP）、（82.95±4.25）%（BTP）、（97.24±5.80）%（OTP）。另外,在细胞培养至第5天检测表明,这3 种TPs显著促进G0/G1期细胞数量的累积,细胞比例分别为（68.52±2.28）%（GTP）、（67.11±1.68）%（BTP）、（59.69±1.35）%（OTP）。RT-PCR分析结果显示TPs可以调控相关脂肪细胞因子的表达。在本研究中,在3T3-L1前脂肪细胞中GTP的减肥作用强于BTP和OTP。TPs的加入上调脂联素的表达从而激活腺苷酸活化蛋白激酶信号通路调控相关脂肪细胞因子的表达,并最终抑制TG的合成与3T3-L1前脂肪细胞的分化。     

GC/MS法同时检测无烟气烟草制品中的1,2-丙二醇、丙三醇和三甘醇

为了准确测定无烟气烟草制品中保润剂的含量,以1,4-丁二醇为内标建立了同时检测无烟气烟草制品中1,2-丙二醇、丙三醇、三甘醇的气相色谱/质谱/选择离子监测(GC/MS/SIM)方法,采用该方法和CORESTA推荐的GC/FID法测定了30个无烟气烟草制品样品,并将2种方法的检测结果进行了比较.结果表明:①该方法的检出限和定量限分别为1.25～2.75 μg/g和4.10-9.05 μg/mL,回收率89.3%～100.3%,相对标准偏差(RSD)1.16%～4.37%;②所测无烟气烟草制品中1,2-丙二醇、丙三醇、三甘醇的含量分别在0.011-40.817,0.024～39.044和0.012～0.099 mg/g之间;③GC/FID法检出13个样品中舍1,2-丙二醇,检出率43.3%;14个样品中含丙三醇,检出率46.7%,其中能够准确定量的有9个样品;未检出三甘醇;④GC/MS/SIM法检出29个样品中含1,2-丙二醇,检出率96.7%,其中能够准确定量的有16个样品;28个样品中含丙三醇,检出率93.3%,其中能够准确定量的有25个样品;21个样品中含三甘醇,检出率70.0%,其中能够准确定量的有11个样品;⑤GC/FID和GC/MS/SIM法共同检出的13个样品中的1,2-丙二醇含量和9个样品的丙三醇含量的皮尔逊(Pearson)相关系数均大于0.99,概率P值均小于显著性水平0.05,配对t检验概率P值均大于显著性水平0.05.结论:两种方法的定量结果一致,但GC/MS/SIM法灵敏度更高,定性更准确,GC/MS/SIM法更适合无烟气烟草制品中1,2-丙二醇、丙三醇、三甘醇的定性定量分析.