冷藏鮰鱼片中优势腐败菌的分离鉴定

为探讨冷藏7 d鮰鱼片中优势腐败菌的种类,将鮰鱼于4℃无菌冷库中取肉后真空包装放置于4℃冰箱中储藏7 d,从腐败鱼肉中分离出3株细菌,编号x1-1、y1-1、y2-1,进行细菌形态学观察、生理生化检测以及16S rDNA序列测定等研究。结果表明所有菌种均为革兰氏阴性短杆菌,且均为兼性厌氧菌,通过测序结果与系统进化树分析显示:x1-1与嗜水气单胞菌亲缘关系最近,达99.9%;y1-1与维氏气单胞菌亲缘关系最近,达99.79%;y2-1异嗜糖气单胞菌亲缘关系最近,达99.8%。以上各项研究结果表明:x1-1为嗜水气单胞菌;y1-1为维氏气单胞菌;y2-1为异嗜糖气单胞菌。综上所述,冷藏鮰鱼片中优势腐败菌为气单胞菌。     

河蚌贮藏鲜度变化及优势腐败菌分析

以菌落总数、挥发性盐基氮(TVBN)与感官评价为指标,对蚌肉在冰藏(0℃)、冷藏(4℃)和室温贮藏(25℃)中的鲜度变化进行定量研究,并分析各贮藏温度下引起河蚌腐败的优势腐败菌。研究表明,0、4℃和25℃贮藏下河蚌的高品质期终点为6、4 d和8 h;货架期终点分别为16、12 d和20 h。0℃冰藏时优势腐败菌是温和气单胞菌(Aeromonas sobria)、少动鞘氨醇单胞菌(Sphingomonas paucimobilis),菌株占比分别为42%、48%。4℃冷藏时优势腐败菌是嗜水气单胞菌(Aeromonas hydrophila),比例为43%。25℃贮藏时优势腐败菌是鹑鸡肠球菌(Enterococcus gallinarum)、温和气单胞菌(Aeromonas sobria),菌株占比分别为46%、51%。     

真空包装冷藏鲟鱼中优势腐败菌的分离、鉴定及生长预测

为监测真空包装鲟鱼在冷藏过程中的优势腐败菌动态变化,通过感官评价和传统选择性培养的方法,对真空包装冷藏鲟鱼中的优势腐败菌进行了分析;同时选择性分离筛选出代表性菌株Q-1和C-1,并基于16S rRNA基因进行了鉴定。结果表明:气单胞菌和肠科菌是真空包装鲟鱼在冷藏过程中的优势腐败菌;菌株Q-1可能为维氏气单胞菌(Aeromonas veronii)新亚种,菌株C-1为路德维希肠杆菌(Enterobacter ludwigii);同时与菌株C-1相比,菌株Q-1的生长延滞期较长,但生长较快,修正的Gompertz模型能够可靠地分别预测菌株C-1和菌株Q-1在真空包装鲟鱼在冷藏过程中生长动态。     

一株鲢鱼肠道中嗜水气单胞菌拮抗菌株的筛选、鉴定及特性

从鲢鱼肠道中筛选嗜水气单胞菌的拮抗菌株,并对生长培养条件进行研究,以期为鲢鱼养殖过程中嗜水气单胞菌病的生物防治提供理论依据。通过对鲢鱼肠道中嗜水气单胞菌拮抗菌株的分离、筛选,并对其中拮抗作用较稳定的菌株进行生理生化、分子鉴定以及生长培养特性优化。经过多次筛选得到了15株拮抗效果较好的目标菌株,其中拮抗作用较稳定的S4为气单胞菌属,是偏碱性革兰氏阴性短杆菌;通过单因素和正交实验表明,S4最佳生长培养条件为1.5 g/d L淀粉,1.5 g/d L牛肉膏,1 g/d L氯化钙,32℃,pH 8.5,接种体积分数3%。研究结果为嗜水气单胞菌的防治及拮抗菌S4的应用提供理论依据。     

湛江地区养殖南美白对虾冷藏期间的菌相分析及优势腐败菌的初步鉴定

将湛江地区养殖的南美白对虾在(4±1)℃条件下进行冷藏保鲜,通过细菌形态观察和生理生化实验对冷藏不同时期的细菌组成进行了定性和定量研究,并对优势腐败菌进行了初步鉴定。结果表明:湛江地区养殖的南美白对虾冷藏初期的细菌总数为2.8×104CFU/g,由10个属的细菌组成,其中有6个属为G-,且它们的数量占细菌总数的80%,其中以黄杆菌和气单胞菌的数量最多,分别占细菌总数的33.3%和13.3%;冷藏的第2天为高品质期,细菌总数为5.1×104CFU/g,黄杆菌(30%)和不动杆菌(16.7%)的数量最多,G-的数量增长到86.7%;随着冷藏时间的延长,细菌种类逐渐减少,货架期终点时细菌总数为3.0×106CFU/g,但只分离到8个属的细菌,其中G-占93.4%,气单胞菌和假单胞菌的数量均增加到16.7%,而黄杆菌减少为26.7%,其他种类的G-均有不同程度的增长。冷藏至第6天,细菌总数高达2.4×107CFU/g,虾肉完全腐败变质,从中分离获得的4个属的优势腐败菌均为G-,它们分别是黄杆菌、希瓦氏菌、假单胞菌和气单胞菌,各自所占比例为28.74%、26.44%、21.84%和16.09%。     

0℃冷藏下三文鱼片菌相变化规律及特定腐败菌的分离鉴定

以冰鲜的三文鱼片为研究对象,利用形态学观察和16S r DNA序列分析法探究其在0℃冷藏过程中腐败菌的变化规律,并结合VITEK2微生物自动鉴定系统对其特定腐败菌进行分离和鉴定。结果表明:0℃冷藏三文鱼片在贮藏初期(第0 d)的腐败菌主要为葡萄球菌属(Staphylococcus sp.)、假单胞菌属(Pseudomonas sp.)、片球菌属(Pediococcus sp.)、微球菌属(Micrococcus sp.)、气单胞菌属(Aeromonas sp.)、肉食杆菌属(Carnobacterium sp.)及部分鞘氨醇单胞菌属(Sphingomonas sp.)。随贮藏时间的延长,腐败菌菌相逐渐单一,葡萄球菌属、片球菌属、微球菌属呈明显的下降趋势;气单胞菌属及鞘氨醇单胞菌属完全消失;肉食杆菌属变化较小,而假单胞菌属呈上升趋势且到贮存后期数量占绝对优势。腐败菌的PCR产物经测序后通过NCBI数据库比对,并经MEGA5.05软件构建系统发育树分析及微生物自动鉴定仪共同鉴定,确定荧光假单胞菌(Ps.Fluorescens)为冷藏三文鱼片的优势腐败菌。     

鮰鱼冷藏过程中气味和新鲜度的变化及相关性

以真空包装与空气包装鮰鱼为研究对象,采用固相微萃取-气相色谱-质谱联用结合电子鼻技术,对鮰鱼4℃冷藏过程中的挥发性成分进行分析,并对不同样品的电子鼻传感器信号进行分析,同时结合感官评分、K值、三甲胺含量、挥发性成分及电子鼻传感器信号进行相关性分析。结果表明：鮰鱼4 ℃冷藏过程中的特征风味物质主要为己醛、壬醛、苯甲醛和1-辛烯-3-醇;电子鼻可以区分不同冷藏时间、不同包装的鮰鱼;感官评定结果表明,鮰鱼冷藏4～5 d时开始腐败,且真空包装组较空气包装组变化缓慢;冷藏鮰鱼三甲胺含量和K值随冷藏时间延长而逐渐上升,真空包装样品新鲜度较空气包装组好;相关性及主成分分析结果表明,苯甲醛、1-辛烯-3-醇可作为表征鮰鱼腐败的挥发性化合物。     

冷藏鲣鱼优势腐败产胺菌分离鉴定及其致腐产胺能力分析

为探究鲣鱼中优势腐败产胺菌的种类,从冷藏的鲣鱼中筛选出3种优势腐败产胺菌,通过16S rDNA分子鉴定技术对菌株进行鉴定为荧光假单胞菌（Pseudomonas fluorescens）、弗氏柠檬酸杆菌（Citrobacter freundii）和嗜水气单胞菌（Aeromonas hydrophila）,将鉴定出的优势腐败产胺菌接种至无菌鱼肉中4 ℃条件下贮藏,通过测定菌落总数和挥发性盐基氮（TVB-N）变化,以腐败代谢产物产量因子（Y_TVB-N/CFU）分析3种优势腐败产胺菌对鲣鱼的致腐能力,并通过样品中生物胺的含量比较3种优势腐败产胺菌的产胺能力。结果表明:在贮藏8 d后接种荧光假单胞菌、弗氏柠檬酸杆菌和嗜水气单胞菌组的菌落总数分别达到8.36、8.27和8.13 lg CFU/g,TVB-N值分别为28.21、30.30和31.29 mg/100 g,3种优势腐败产胺菌对鲣鱼的致腐能力大小为嗜水气单胞菌>弗氏柠檬酸杆菌>荧光假单胞菌。接种荧光假单胞菌组、弗式柠檬酸杆菌组和嗜水气单胞菌组的组胺含量分别为196.23、83.43和261.22 mg/kg,3组样品中总生物胺含量为嗜水气单胞菌>荧光假单胞菌>弗氏柠檬酸杆菌。综合比较得出,嗜水气单胞菌对4 ℃冷藏鲣鱼致腐产胺能力最强,本研究增加对冷藏鲣鱼中优势腐败产胺菌种类的了解并提供了部分理论基础。     

PCR结合生理生化鉴定对冷藏带鱼主要细菌菌相组成分析

将传统的微生物生理生化鉴定技术与16S rDNA分子生物学方法相结合,并通过系统发育树分析,对冷藏带鱼贮藏期间的主要细菌组成比例进行研究。样品在(4±1)℃贮藏,分别于不同贮藏时间取样,对各个阶段的微生物菌落进行分离纯化、形态观察和部分生理生化鉴定,对最终得到的细菌纯培养物直接提取DNA,作16SrDNA PCR扩增,产物经测序后于NCBI与已知序列进行相似性比对,最终得到13株具有典型特征的纯菌株。结果得出,冷藏带鱼在货架期终点时,其主要微生物的种类与所占比例依次为:腐败希瓦氏菌(Shewanella putrefaciens 34.7%)、荧光假单胞菌(Pseudomonas fluorescens 14.5%)、松鼠葡萄球菌(Staphylococcus sciuri 10.2%)、嗜水气单胞菌(Aeromonas hydrophila 8.2%)、弧菌(Vibrio rumoiensis 6.1%)、恶臭假单胞菌(Pseudomonas putida6.1%)、绿色气球菌(Aerococcus viridans 4.1%)、金黄色葡萄球菌(Staphyloccocus aureus 4.1%)、蜡样芽孢杆菌(Bacillus cereus 4.0%)、铜绿假单胞菌(Pseudomonas aeruginosa 2.0%)、嗜冷杆菌(Psychrobacter sp.2.0%)、成团肠杆菌(Enterobacter agglomerans 2.0%)、约氏不动杆菌(Acinetobacter johnsonii 2.0%),其中腐败希瓦氏菌为带鱼冷藏期间的主要优势腐败菌,假单胞菌在带鱼冷藏过程中的所占比例大小依次为荧光假单胞菌>恶臭假单胞菌>铜绿假单胞菌。     

酒精浸泡冷藏鲜印度块菌货架期评估、挥发性物质与细菌群落变化及其相关性

为探索延长鲜印度块菌货架期的简单有效的冷藏保鲜技术,本实验以我国产量最高的鲜印度块菌为对象,对其在4℃冷藏过程中的感官指标、菌落总数、主要营养组分质量损失率和挥发性物质含量进行研究,进一步采用高通量测序技术分析细菌群落变化,探究对鲜印度块菌香气贡献程度较大的挥发性物质和主要营养组分与细菌群落变化的相关性,通过16S rRNA测序测定鲜印度块菌冷藏过程中的优势腐败菌。结果表明,采用体积分数75%食用酒精速泡、真空包装后4℃冷藏,鲜印度块菌货架期可到30 d;在冷藏的38 d内鲜印度块菌的总糖、总蛋白、粗纤维相对含量与0 d相比明显下降,质量损失率明显上升,挥发性芳香物质含量明显下降,挥发性异味物质含量也明显增加。经高通量测序的细菌群落分析结果表明鲜印度块菌冷藏期间的优势菌门为Proteobacteria、Bacteroidetes、Actinobacteria、Firmicutes和Verrucomicrobia。基于属水平的冗余分析结果表明,在0～15 d营养组分（总糖、总蛋白和粗纤维）相对含量和特征性芳香物质（1-辛烯-3-醇、正戊酸正戊酯、乙酸乙酯、3-甲基丁酸乙酯、2-辛酮、1-...     

肉制品中亚硝酸盐测定能力验证

为加强肉制品安全监督管理,进一步提升检验检测机构对肉制品中亚硝酸盐含量的检测能力,确保提供的检测数据准确、可靠,组织实施肉制品中亚硝酸盐含量检测能力验证计划。制备含亚硝酸钠的鸡肉火腿肠,向42家实验室随机发放样品,在规定时间内回收检测数据。对检测结果进行稳健统计分析,以Z值评价各个实验室的检测能力,并对非满意结果的实验室通过查阅原始记录进行技术分析。结果表明：制备的能力验证样品均匀、稳定,满足能力验证计划要求;42家实验室总体结果满意率为81.0%,表明实验室对肉制品中亚硝酸盐含量的检测能力较好;部分实验室发现检测过程中所存在的问题,并加以改正,使自身检测能力进一步提高。     

Foodborne pathogenic bacteria in prepackaged fresh retail portions of farmed rainbow trout and salmon stored at 3 degrees C

Twelve lots of fresh unskinned fillets of rainbow trout (Oncorhynchus mykiss) and 10 lots of fresh sliced salmon (Salmo salar) prepacked in trays wrapped with an oxygen-permeable film were obtained immediately after packing from two supermarkets having in-plant facilities for packaging wet fish. During storage at 3 degrees C, Listeria innocua was detected in eight lots of trout fillets after 4 days storage. L. monocytogenes was recovered from a single lot also contaminated with L. innocua. Initial numbers of aeromonads were significantly (p < 0.05) lower in trout fillets (3.35 +/- 0.62 log cfu g(-1)) than in salmon slices (4.20 +/- 0.89 log cfu g(-1)). In both fish products, these bacteria significantly (p < 0.05) increased up until spoilage. Most Aeromonas spp. isolates from trout fillets were assigned to A. veronii biovar sobria HG8 (hybridisation group 8), A. caviae HG4, A. eucrenophila HG6, A. hydrophila HG1 and A. veronii biovar veronii HG10. Strains of HG12 (A. schubertii), HG4 and HG8 formed the majority of aeromonads recovered from salmon slices.     

Reduction of endogenous bacteria associated with catfish fillets using the Grovac process

Fresh catfish (Ictalurus punctatus) fillets are known to be contaminated with a large number of spoilage and pathogenic bacteria. The Grovac method, a new patented (U.S. 5,543,163) process, was evaluated for its efficacy in reducing the number of pathogens and spoilage microorganisms associated with food. This process involves using a processing solution containing ascorbic acid (AA) and sodium chloride (NaCl), vacuum, and tumbling. A total of 51 bacterial isolates were isolated and identified from whole catfish and catfish fillets using both selective and nonselective media, phenotypic tests, and the Vitek identification system. Psychrotrophic foodborne pathogens included: Aeromonas hydrophila, Escherichia coli, Listeria sp., Plesiomonas shigelloides, Proteus sp., Staphylococcus aureus, and Vibrio parahaemolyticus. High aerobic plate counts (2.6 x 10(7) CFU/g) for catfish fillets indicated that fillets were heavily contaminated during processing of catfish. The Grovac process showed that various treatment combinations of AA and NaCl resulted in a 1.2 to 2.3 CFU/g log reduction of microbial counts associated with catfish fillets. The effectiveness of the process may be related to the synergistic effect of tumbling, AA, NaCl, and vacuum. These results suggested that the Grovac process could be used as an alternative processing procedure to reduce microbial populations associated with catfish fillets and may be useful to improve the shelf-life and food safety of the product. Microbiological data from this study will be used for the development of a hazard analysis for the implementation of the hazard analysis critical control point program for processed catfish fillets.     

新疆椒麻鸡中腐败细菌的分离与鉴定

为研究新疆椒麻鸡中腐败细菌的种类和特性,采用平板划线法分离腐败菌。利用生理生化实验和16S rDNA序列分析方法,对从椒麻鸡中分离的31 株腐败菌进行鉴定。结果表明：所筛选的31 株菌中,肠杆菌20 株、葡萄球菌3 株、气单胞菌2 株、乳酸菌4 株、溶酪大球菌2 株;新鲜椒麻鸡中的主要腐败菌为溶酪大球菌、葡萄球菌和肠杆菌;低温5 ℃贮藏条件下腐败样品中的主要腐败菌为乳酸菌、葡萄球菌和肠杆菌;常温25 ℃贮藏条件下腐败椒麻鸡中的腐败菌主要为肠杆菌、气单胞菌、葡萄球菌和乳杆菌。     

冷藏小龙虾优势腐败菌的筛选鉴定及细菌群落结构和多样性分析

以4℃贮藏小龙虾货架期终点筛选武汉孝感小龙虾中优势腐败菌,同时结合高通量测序方法,进一步确定小龙虾中优势腐败菌,并对小龙虾贮藏过程中菌相变化、微生物结构及其多样性进行分析。结果表明：传统培养法从腐败小龙虾中共筛选出10 株腐败菌,包括波罗的海希瓦氏菌（Shewanella baltica）、维氏气单胞菌（Aeromonas veronii）、普通变形杆菌（Proteus vulgaris）等;在属水平上,高通量测序法最终确定气单胞菌属（Aeromonas sp.）及希瓦氏菌属（Shewanella sp.）是小龙虾腐败末期相对丰度最大的2 种优势腐败菌;同时,高通量测序结果表明,随着贮藏时间的延长,小龙虾微生物群落丰富度、多样性及均匀度不断下降,这与腐败末期优势腐败菌的大量增长及生长繁殖过程中抑制其他种类微生物有关。     

Effect of packaging atmosphere on the microbial attributes of pearlspot (Etroplus suratensis Bloch) stored at 0-2 degrees C

Effect of packaging atmosphere (air and under different modified atmospheres (MAs), 40% CO2/60% O2, 50%/50% O2, 60% CO2/40% O2, 70% CO2/30% O2 and 40% CO2/30% O2/30% N2) on the microbial and biochemical attributes of fresh pearlspot (Etroplus suratensis Bloch) stored at 0-2 degrees C was investigated. Trimethylamine nitrogen (TMA-N) and thiobarbituric acid (TBA) values remained lower than the proposed acceptability limits throughout the storage period. Results demonstrated that storage of pearlspot under air and MA 40% CO2/30% O(2)/30% N(2) resulted in growth of Enterobacteriaceae, Aeromonas and H(2)S-producing bacteria including Shewanella putrefaciens, while all other packaging atmospheres did not allow multiplication of Enterobacteriaceae and Aeromonas within 3 weeks. Aeromonas spp. identified were Aeromonas hydrophila, Aeromonas veronii biovar sobria and A. veronii biovar veronii. Significant reduction (p<0.01) was noticed in Aeromonas population of pearlspot stored under MA 60% CO2/40% O2 and 70% CO2/30% O2. A delay of growth of Pseudomonas below 5.0log(10)cfug(-1) was observed during the 15th day of storage at 0-2 degrees C under modified atmosphere packaging (MAP) conditions. Growth of faecal streptococci was significantly inhibited in all the packaging atmospheres at 0-2 degrees C during the entire storage period. Survival of coagulase positive Staphylococci (<50cfug(-1)) in low numbers was noticed during storage in all the packaging atmospheres. Clostridium botulinum toxin was not detected. All the packaging atmospheres did not allow multiplication of sulphite-reducing clostridia at 0-2 degrees C during the entire storage period. Packaging in MA 60% CO2/40% O2 resulted in the inhibition of growth of Aeromonas and Enterobacteriaceae, and the slowest growth of psychrotrophic bacteria, H(2)S-producing bacteria, including Shewanella putrefaciens and Pseudomonas and extended microbiological shelf life to 9-10 days. This study confirms the survival of potentially pathogenic A. hydrophila, A. veronii biovar sobria and A. veronii biovar veronii capable of growth at low temperature in pearlspot stored under MA.     

Potassium acetate and potassium lactate enhance the microbiological and physical properties of marinated catfish fillets

Sodium or potassium salts such as lactate and acetate can be used to inhibit the growth of spoilage bacteria and food-borne pathogens, and thereby prolong the shelf-life of refrigerated seafood. However, minimal information is available regarding the combined effects of potassium salts (acetate and lactate) with an agglomerated phosphate blend on the quality and safety of refrigerated catfish fillets. The objective of this study was to determine the microbiological and quality characteristics of marinated catfish fillets treated with organic acid salts. Catfish fillets were vacuum-tumbled with a brine solution with and without the added organic acid salts, at 10% over initial, raw weight prior to tray-packing and storage at 4 °C for 14 d. Fillets were evaluated for yields, color, pH, tenderness, consumer acceptability, and shelf-life. No differences (P > 0.05) existed among the treated and untreated fillets with regards to solution pick-up and pH, but all treated fillets increased (P < 0.05) cooking yields and Intl. Commission on Illumination (CIE) a* values, and decreased (P < 0.05) CIE L* and b* values in the catfish fillets when compared to the untreated fillets. The fillets treated with a combination of potassium acetate and potassium lactate had lower (P < 0.05) psychrotrophic plate counts and lower spoilage scores than the control treatments on days 7, 10, and 14. In addition, consumers preferred (P < 0.05) treated catfish fillets (fried) with respect to appearance, flavor, and overall acceptability over the negative control. In conclusion, the combination of potassium acetate and potassium lactate enhanced sensory quality and extended the shelf-life of refrigerated catfish fillets.     

真空包装冷藏大菱鲆鱼片的货架期及其优势腐败菌群体感应检测

本研究以真空包装冷藏大菱鲆鱼片为研究对象,通过感官、微生物和化学指标评价其货架期,利用16Sr DNA方法对其贮藏初期和末期的微生物分布进行鉴定,并采用报告菌株法检测大菱鲆鱼片贮藏过程中两种类型群体感应信号分子(AHLs和AI-2)的产生情况,检测其优势腐败菌的群体感应类型。结果显示,大菱鲆鱼片在真空包装冷藏环境下可以贮藏9 d,此时细菌总数达6.87 logCFU/g,挥发性盐基氮含量为28.70 mg/100 g。对比初始和末期微生物种类变化,真空包装大菱鲆鱼片冷藏过程中的优势腐败菌主要为希瓦氏菌,其次为肠杆菌科细菌。群体感应信号分子AHLs和AI-2含量随贮藏时间延长而增加。贮藏末期分离的希瓦氏菌均能分泌AI-2信号分子,分泌能力因菌株而异。利用基因手段可从希瓦氏菌中扩增到AI-2的分泌基因lux S,表明lux S在产AI-2菌株中具有一定保守性,可能参与希瓦氏菌的腐败进程。