Characterization of Staphylococcus xylosus and Staphylococcus carnosus isolated from Slovak meat products

The aims of this study were to isolate, identify and characterize the population of coagulase-negative staphylococci in different types of Slovak traditional sausages and to determine the metabolic properties of selected Staphylococcus xylosus and S. carnosus strains for the selection of potential starter cultures to use in the processing of sausages. The strains were tested for lactic acid production, survival in the presence of bile and sensitivity to antibiotics. Bacteriocin production, adhesion ability as well as biogenic amine (BA) production by isolates were also analysed. Most of the isolates were identified as S. xylosus and S. carnosus. Lactic acid values ranged from 0.40 to 1.03 mmol/l and strains survived in the presence of 1% bile. Most of the strains studied were sensitive to all antibiotics. Two strains, S. xylosus SO3/1M/1/2 and S. carnosus SO2/F/2/5 inhibited Listeria innocua and Pseudomonas sp. S. xylosus strains did not produce any BA, while S. carnosus SO2/F/2/5 did. S. xylosus SO3/1M/1/2 and S. carnosus SO2/F/2/5 appeared as the most adhesive strains. S. xylosus SO3/1M/1/2 with antimicrobial activity against Enterococcus avium EA5, L. innocua LMG13568 and Pseudomonas sp. SO1/1M/1/4, adhesion ability and free BA production could be used as starter culture in sausage manufacture.     

Influence of Beet Sugar,Calcium Lactate,and Staphylococcus xylosus (with Nitrate Reductase Activity) on the Chemical,Microbiological, and Sensorial Properties of Persian Uncured Frankfurters

Abstract: This study investigated the effects of beet sugar (BS), calcium lactate (CL), and Staphylococcus xylosus (SX) on the general characteristics of uncured frankfurters. Minimum residual nitrite was observed in samples with a high level of BS and S. xylosus (8log₁₀ l0 viable cfu/g) after 2 mo. There was an increase in total aerobic counts in frankfurters after 4 wk. Samples containing higher BS and CL had no Clostridium perfringens at storage time. Histamine content was similar for all treatments except SX. No differences were observed in textural properties among the treatments. Water activity was decreased significantly in treatment CL1+BS2. The results indicated that samples treated with BS or CL had (P < 0.05) taste and sensory attributes considered appropriate by a tasting panel. Practical Applications: Frankfurters are among the most popular meat products in many countries. Health problems (such as cancer) associated with the overconsumption of cured meat have led to searches for new, alternative methods of preservation. Hurdle technology (combinations of beet sugar, calcium lactate, and Staphylococcus xylosus in different concentrations) can be developed for frankfurter processing. Results of this research will be useful for meat manufacturers; however, any new process may affect the strategies chosen for marketing these products.     

一氧化氮对采后油菜品质与活性氧代谢相关酶的影响

NO是植物体内重要的信号分子,其在采后保鲜方面的应用是近年研究的焦点。本研究以油菜(Brassicarapa L.ssp.chinensis,cv.Huaguan)为试材,使用一氧化氮(NO)供体100μmol/L硝普钠(SNP)-0.04mp真空渗透处理2min,贮藏于25℃相对湿度(RH)70%的环境中。在贮藏后0、1、2、3、4d分别测定油菜的腐烂率及黄变率;并测定贮藏过程中超氧化物歧化酶(SOD),过氧化氢酶(CAT),抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)的活性。结果表明,SNP处理显著降低了采后油菜的黄变率及腐烂率;提高了油菜贮藏期间SOD、APX的活性,降低了1～3d的GR活性;贮藏1d时提高了CAT的活性,2d之后抑制CAT活性。说明100μmol/L SNP处理提高了油菜采后品质,并可能通过提高贮藏期前期CAT活性和整个贮藏期间SOD及APX的活性延缓油菜采后品质的下降。     

金银花提取物修护外源NO致NIT-1胰岛β细胞损伤

将金银花提取物(Honegsukle Flower P.E,HFPE)作用于受外源性一氧化氮(NO)损伤的NIT-1胰岛细胞,测定细胞存活率、超氧化物歧化酶(SOD)活力、丙二醛(MDA)释放量、一氧化氮(NO)释放量、胰岛素分泌5项生理指标,以研究HFPE对NIT-1胰岛细胞的保护作用。研究结果表明:HFPE其浓度在0.5 g/L~0.000 5 g/L范围内可减少NO、MDA释放,提高SOD活力水平,提高细胞存活率,增加胰岛素的分泌。     

一氧化氮(NO)熏蒸提高哈密瓜果实采后贮藏的耐冷性

为了探究一氧化氮(nitric oxide,NO)熏蒸对哈密瓜采后耐冷性的影响。以"西州密25号"为试材,用60μL/L NO气体熏蒸3 h,分别在(1±0.5℃)、(3±0.5℃)、(5±0.5℃)贮藏。统计冷害发病率和冷害指数,测定过氧化氢酶(catalase,CAT)、过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)活性、超氧阴离子(superoxide anion,O_2~ˉ·)产生速率、过氧化氢(hydrogen peroxide,H_2O_2)含量的变化。结果表明:NO处理能提高SOD、POD活性,抑制O_2~ˉ·产生速率。1℃条件下,果实冷害发病率最高,NO处理能减轻冷害的发生;3℃条件下,对照果实后期出现冷害,但NO处理果实无冷害发生;5℃条件下,果实后期出现冷害和病害症状,NO处理能减轻果实的冷害和病害。说明NO能够提高抗氧化相关酶的活性,抑制活性氧(reactive oxygen species,ROS)的积累,提高果实的耐冷性;3℃条件下,NO熏蒸既能有效维持果实的贮藏品质又能抑制果实冷害的发生。     

一氧化氮处理对甜瓜果实采后病害的控制及活性氧代谢的作用

为探究一氧化氮处理对甜瓜果实采后病害的控制及活性氧代谢的关系。以新疆‘西州蜜25号’甜瓜为试验材料,将60μL/L的一氧化氮(nitric oxide,NO)气体充入密封箱熏蒸3 h,密封箱中不充入NO气体密封3 h的处理作为对照,放置12 h后损伤接种胶孢链格孢(Alternaria alternata),置于常温条件下贮藏。定期测定甜瓜果实病斑直径和病斑深度及与活性氧代谢相关的过氧化氢酶(catalase,CAT)、过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)及超氧阴离子(superoxide anion,O_2~-·)的产生速率和过氧化氢(hydrogen peroxide,H_2O_2)的含量。结果表明:NO处理能明显降低贮藏期间甜瓜果实的病斑直径和病斑深度,诱导贮藏前期O_2~-·的产生速率加快和H_2O_2含量的快速积累,明显提高贮藏期间甜瓜果实SOD、CAT和POD的活性。说明NO处理诱导果实采后抗病性与调节果实的活性氧代谢密切相关。     

NO处理对采后莲雾果实絮状绵软进程的影响

探明NO处理对采后莲雾果实成熟衰老过程中絮状绵软进程的影响。以台湾‘黑珍珠’莲雾果实为材料,用不同用量（5、10、20 μL/L）外源NO熏蒸处理莲雾果实,以0 μL/L NO熏蒸处理作对照,研究其对采后莲雾果实生理及品质、细胞壁代谢及木质素代谢的影响。结果表明：与对照相比,不同用量外源NO熏蒸处理能够显著抑制莲雾果实贮藏过程中絮状绵软指数及质量损失率的上升,保持果实硬度,延缓果实纤维素含量的下降,10 μL/L NO显著降低了多聚半乳糖醛酸酶（PG）、果胶甲酯酶（PME）、β-半乳糖苷酶（β-Gal）活性。同时,不同用量外源NO熏蒸处理能够延缓果实木质素含量的上升,10 μL/L NO显著降低了苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）、4-香豆酸辅酶A连接酶（4-CL）活性。相关性分析表明,絮状绵软指数与质量损失率、果肉硬度、纤维素含量和木质素含量之间均存在显著相关性,且与原果胶含量呈极显著负相关,与可溶性果胶含量呈极显著正相关。NO处理能够抑制细胞壁类物质降解酶活性,延缓采后莲雾果实絮状绵软进程,延长果实贮藏期,其中以10 μL/L NO处理效果最好。     

多巴胺磁性纳米粒子对Nisin的吸附特性

本实验旨在通过系统研究多巴胺磁性纳米粒子（iron oxide nanoparticles coated with polydopamine,IONPs@pDA）对乳酸链球菌素（Nisin）的吸附特性,为新型杀菌材料Nisin-IONPs@pDA耦合体的制备提供理论依据及数据支持。实验测定了不同温度和时间下多巴胺磁性纳米粒子对Nisin的吸附量,并进行吸附动力学和吸附等温线拟合。结果显示：多巴胺磁性纳米粒子对Nisin的吸附行为符合Lagergren准二级吸附动力学模型和Freundlich吸附等温模型,说明吸附过程主要受化学吸附的控制,且为发生在异构表面的多层吸附过程。热力学计算结果表明,多巴胺磁性纳米粒子对Nisin的吸附是一个吸热过程,Nisin的吸附量随着温度的升高而增大。结论：多巴胺磁性纳米粒子是一种有效的Nisin吸附磁性材料,可用于制备新型复合杀菌材料。     

Molecular and technological characterization of Staphylococcus xylosus isolated from naturally fermented Italian sausages by RAPD, Rep-PCR and Sau-PCR analysis

Coagulase-negative cocci (CNC) are important microorganisms in fermented sausages because they release lipases and proteases that are able to free short-chain fatty acids and peptides and aminoacids, respectively, that are responsible for the aroma of fermented sausage. The purpose of this study was to characterize Staphylococcus xylosus strains isolated from naturally fermented sausages, produced in three different processing plants in the Friuli Venezia Giulia region in the Northeast of Italy. Two hundred and forty-nine strains of S. xylosus were identified by species-specific PCR and subjected to molecular and technological characterization. RAPD-PCR with primer M13, Rep-PCR using primer (GTG)(5) and Sau-PCR with primer SAG(1) were used for the molecular analysis, while the capability of the strains to grow at different temperatures and in the presence of NaCl and their lipolytic and proteolytic activity were tested in order to define the technological characteristics. The results obtained allowed us to differentiate strains coming from different plants, thereby admitting the presence of strains that are plant-specific.     

环介导恒温扩增法(LAMP)检测金黄色葡萄球菌

建立一种能够快速准确地检测金黄色葡萄球菌的LAMP方法。根据金黄色葡萄球菌的femA基因设计了引物,然后进行LAMP反应条件的优化、特异性和灵敏度的检测并与实际样品进行检出率的比较。LAMP方法特异性好,最佳反应温度为61℃,只对金黄色葡萄球菌进行扩增;灵敏度高,金黄色葡萄球菌的检测灵敏度为8～9cfu/mL时仍能检出。LAMP方法检测金黄色葡萄球菌特异性强、灵敏度高、时间短且操作简便,有望成为快速检测金黄色葡萄球菌的新方法。     

一次过柱法快速分离纯化驴心肌中高铁肌红蛋白

肌红蛋白及其存在形式是决定肉和肉制品色泽的重要因素,高铁肌红蛋白为其重要存在形式之一。以马科动物-驴的心肌为原料,采用70%～100%饱和硫酸铵分段盐析法粗提肌红蛋白,加入铁氰化钾氧化后透析,经CM-Sephadex Fast Flow阳离子交换柱层析进行一步纯化。SDS-PAGE电泳和紫外/可见光光谱扫描结果表明,仅通过一次柱交换,即可得到电泳纯的高铁肌红蛋白,为快速提取、纯化高铁肌红蛋白提供了一条途径。     

一氧化氮熏蒸处理抑制冬枣贮藏过程中的乙醇代谢及异味产生

本研究以冬枣果实为试验材料,用20μL/L一氧化氮(NO)气体熏蒸冬枣果实3 h,置于0℃、相对湿度为90%～95%的冷库贮藏75 d,定期测定冬枣果实挥发性物质、乙醇代谢产物及关键酶活性的变化。结果表明,贮藏期冬枣果实共鉴定出47种挥发性物质,主要包括醇类、醛类、酯类、酸类、酮类。在采收当天、以及对照组和NO熏蒸处理组贮藏45 d和75 d后,分别检测出24、37、38、45和46种挥发性成分。贮藏期间果实醛类物质的变化多以先上升后下降为主,醇类和酯类物质的种类和含量大致主要呈增加趋势,NO对于醛类物质的作用因贮藏时间和醛类种类有所不同,而NO能有效抑制醇类和酯类含量增加。同时,NO处理显著抑制了丙酮酸脱羧酶(PDC)和乙醇脱氢酶(ADH)的活性,进而抑制了果实贮藏期间乙醇和乙醛含量的积累。说明NO处理能较好地保持冷藏果实挥发性物质的含量,延缓异味产生。     

猪源金黄色葡萄球菌lsa(E)基因的流行性研究

多重耐药菌的传播对食品安全和人们健康造成严重威胁。为研究介导细菌对林可胺类、截短侧耳素类及链阳菌素A类耐药的lsa(E)基因及其基因簇在猪源金黄色葡萄球菌中的流行情况,对厦门三个猪场分离得到的29株金黄色葡萄球菌进行lsa(E)基因检测,用overlapping PCR对lsa(E)基因阳性菌株进行耐药基因簇的扩增,并用药敏纸片法检测菌株对抗菌药物的敏感性。结果表明,29株金葡菌全部携带lsa(E)基因,且aad E-spc-lsa(E)-lnu(B)-tnp基因簇检出率为96.6%(28/29)。药物敏感实验结果显示,29株lsa(E)基因阳性的金葡菌对克林霉素、克拉霉素、庆大霉素的耐药率分别为100.0%、100.0%、72.4%,对复方新诺明、四环素、环丙沙星的耐药率均为96.6%。未发现有对苯唑西林、喹奴普汀/达福普汀和利奈唑胺耐药的菌株。细菌多重耐药率为100.0%,且耐药谱主要为PEN-GEN-TET-CLA-SXT-CLI-CIP-MXF。研究旨在为临床治疗食源性致病菌引起的食品安全问题和控制多重耐药菌随食品链的传播提供一定的科学依据。     

牛奶源金黄色葡萄球菌血清型、毒力基因及PFGE分型

目的：研究生鲜牛奶中金黄色葡萄球菌分离、荚膜多糖血清型分布、毒力基因携带及脉冲场凝胶电泳（pulsed field gel electrophoresis,PFGE）分型情况。方法：从河南省4?个地区奶牛养殖场采集生鲜牛奶样品按照国标法进行分离,扩增耐热核酸酶基因nuc鉴定金黄色葡萄球菌,利用聚合酶链式反应方法测定荚膜多糖血清型和毒力基因携带情况,采用PFGE分析菌株间的相关性和遗传关系。结果：从350?份生鲜牛奶样品中分离鉴定到80?株金黄色葡萄球菌,分离率为22.86%。荚膜多糖血清型测定发现,cap5（60%）是流行血清型。从这些阳性菌株中,发现有62?株（77.5%）携带有毒力基因,毒力基因set、hlb、hld、lukED、ebp、clfA和clfB,检出率分别为40.00%、51.25%、57.50%、60.00%、58.75%、57.50%和58.75%。此外,47?株（58.75%）菌携带不少于6?个毒力基因,流行的毒力基因谱型为set-hla-hlb-hld-lukED-cna-ebp-clfA-clfB。PFGE结果显示,获得72?株菌的PFGE图谱,按90%的相似性可分为12?个簇和46?种PFGE型。D簇（3?种PFGE型）、G簇（3?种PFGE型）和J簇（5?种PFGE型）菌株中均检出一定基因类型的毒力基因,表明河南地区生鲜牛奶中金黄色葡萄球菌毒力基因广泛存在于多种PFGE型别中。结论：生鲜牛奶均有一定程度的金黄色葡萄球菌污染,多数菌株携带毒力基因,且毒力基因的类型较为复杂,这对消费这些牛奶的人群构成潜在的健康威胁。PFGE分型菌株主要以克隆形式进行传播,且克隆型具有多样性和差异性,故临床应加强生奶及乳品血清型、毒力基因检测及分子分型研究。     

Recovery of Staphylococcus aureus in Gray Mugil cephalus Roe (Bottarga): Investigation by an Integrated Cultural/Molecular Approach

In the Mediterranean area, salted and dried roe from the gray Mugil cephalus “bottarga” represent a speciality food with great commercial value. Bottarga is currently produced by a traditional handmade process and, the risk of human bacterial contamination during its manufacturing is still unknown; in this perspective the foodborne pathogen Staphylococcus aureus could potentially contaminate this product due to poor sanitation or bad handling during processing. The aim of this work is: to evaluate the contamination level of foodborne pathogens at different product manufacturing stages and, in addition, to describe a fast and realizable method for the rapid detection of S. aureus in bottarga samples in the field. A cultural procedure was initially used to investigate the occurrence of S. aureus and the other main foodborne pathogens in bottarga samples at the different manufacturing stages (from roe to final product). In addition, a molecular approach was used to rapidly determine the presence of total bacteria, S. aureus, and its potential toxigenicity. Of the 194 specimens analyzed, we identified: Clostridium perfringens, Enterococcus spp. and Enterobacteriaceae. However, some samples resulted as being contaminated with S. aureus (4% in roe and 8.7% in the final product). During the bottarga manufacturing process, we observed an increase in pathogen levels (from 10² to 10⁵ CFU/g) in contaminated samples, and entA and entB genotypes were identified. Reconstruction experiments suggest that the fresh roe and the bottarga (not completely dried) could represent a risk for the contamination and growth of pathogen bacteria.     

乌鸡低聚肽亚铁螯合物的分离纯化与结构鉴定

从乌鸡中酶解得到乌鸡低聚肽,然后与亚铁螯合制备乌鸡低聚肽亚铁螯合物,螯合率为84.76±0.12%。乌鸡低聚肽亚铁螯合物具有较高的蛋白质含量,高达54.64±1.03%。并且分子量较低,其中分子量小于1000 u的占85.50%。通过扫描电镜、红外光谱对乌鸡低聚肽亚铁螯合物的结构进行分析,结果表明乌鸡低聚肽亚铁螯合物是一种新型的铁螯合物。体外稳定性研究表明乌鸡低聚肽亚铁螯合物具有一定的热稳定性、酸碱稳定性和体外消化稳定性。通过反相高效液相色谱对乌鸡低聚肽亚铁螯合物进行分离纯化,选择一个主要的组分进行收集,然后利用质谱仪进行质谱分析。从乌鸡低聚肽亚铁螯合物的主要组分中鉴定出一个五肽,氨基酸序列为Thr-Ser-Gly-Met-Pro。乌鸡低聚肽亚铁螯合物可作为一种铁补充剂用于食品添加剂、营养物及医药制品中。     

Purification and characterisation of antioxidant and nitric oxide inhibitory peptides from Tilapia (Oreochromis niloticus) protein hydrolysate

Nitric oxide (NO)‐inhibitory and antioxidative activities of tilapia hydrolysates were prepared using ultrasound pretreatment at 70 W for 30 and 45 min, respectively, followed by Flavourzyme hydrolysis for 1 h. Both hydrolysates were fractionated using size exclusion chromatography on Sephadex G‐25 column and purified by RP‐HPLC. The amino acid sequence of the most potent and purified fractions was determined using LC/MS/MS. The antioxidant peptide (KAFAVIDQDKSGFIEEDELKLFLQNFSAGARAGDSDGDGKIGVDEFAALVK, MW: 6334.49 KDa) and NO‐inhibitory peptide (AFAVIDQDKSGFIEEDELKLFLQNFSAGARAGDSDGDGKIGVDEFAALVK, MW: 6309.49 Da) produced no cytotoxicity in RAW264.7 macrophage cell lines at the concentration of 100 mg mL^?1. The purified peptides at the concentration 100 μg mL^?1 possessed antioxidative and NO‐inhibitory activities 83.0 ± 1.1% and 40.9 ± 0.2%, respectively, which were about 100 times those of their counterpart crude hydrolysates.     

响应面法优化水溶性大豆多糖- 铁(Ⅱ)配合的合成工艺

优化水溶性大豆多糖-铁(Ⅱ)配合物的合成工艺,在单因素试验基础上,应用响应面法二次回归正交旋转组合试验设计,分析水溶性大豆多糖与催化剂柠檬酸三钠的质量比、pH值、反应时间及温度对配合物铁含量的影响,建立相应的预测模型。方差分析结果表明：质量比、pH值对铁含量有显著影响。优化所得的较优工艺参数为水溶性大豆多糖与柠檬酸三钠质量比1.89:1、pH3.89、反应时间1.56h、温度60.6℃。对应的铁含量的预测值为23.08%,实际平均值为21.89%。结果表明：应用响应面法所得到的水溶性大豆多糖-铁(Ⅱ)配合物的合成工艺参数是可行的。     

Improvement of Fish-sauce Odor by Treatment with Bacteria Isolated from the Fish-sauce Mush (Moromi) Made from Frigate Mackerel

ABSTRACT: A bacterium, which was isolated from fish-sauce mush (moromi) of frigate mackerel and identified as Staphylococcus xylosus , could change notes of an odor in fish sauce made in Thailand. Volatile compounds of the fish sauce after incubation at 32 °C for 24 d with the cultured bacterium were analyzed by gas chromatogra-phy. Although the contents of 2-ethylpyridine and dimethyl trisulfide were reduced during the incubation, 4 volatile compounds, including 2-methylpronal, 2-methylbutanal, 2-pentanone, and 3-(methylthio)propanal, showed no significant quantity changes. On the other hand, although 3-methyl-1-butanol and 2,6-dimethylpyrazine were markedly increased, 3-methylbutanoic acid was slightly increased. As a result of sensory evaluation, fishy, sweaty, fecal, and rancid notes of the fish sauce treated with the bacterium were all weaker than those of the nontreated fish sauce. No changes were found with respect to burnt, cheesy, meaty, and ammoniacal notes between fish sauce treated with the bacterium and that without treatment. Taken together, the odor of fish sauce was much improved by treatment with the isolated bacterium.