Changes of bacterial diversity and main flora in chilled pork during storage using PCR-DGGE

This study was designed to explore the bacterial diversity and the main flora in chilled pork by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Longissimus muscle was removed from pork carcasses at 24 h postmortem. The muscle was tray- and vacuum-packaged at 4 degrees C for 2, 4, 7 days to extract the bacteria total DNA, respectively. The results indicated that the bacterial diversity of chilled pork decreased with storage time regardless of packaging method. Nine types of bacteria were identified, including Arthrobacter sp., Enterococcus sp., Staphylococcus sp., Moraxella sp., Pseudomonas sp., Lactobacillus sp., Aeromonas sp., Acinetobacter sp., Brochothrix thermosphacta. For tray-packaged pork, Pseudomonas sp. and B. thermosphacta were the dominant micro-organisms. The differences in the species found were related with the presence of Lactobacillus sp. in vacuum-packaged meat. The results of the present study might be useful to study the changes of the contaminating bacteria and their characteristics in chilled pork.     

Palatability and Texture of Ground Meat Patties Made with Varying Amounts of Pork and Turkey

Meat mixtures were prepared containing pork lean and turkey lean in ratios of 0/100, 33/67, 100/0 and a constant fat content. Blends were prepared with and without a commercial seasoning-antioxidant mixture (SA) and evaluated after 0, 3, 6 or 9 days (4°C) storage or 0, 14 or 28 days (-20°C) storage. Flavor intensity scores for blends containing up to 67% turkey were not different (P > 0.05); however, 100% turkey patties had lower flavor intensity scores. Off-flavor and firmness scores were not affected by blend for products containing between 33% and 100% pork lean, while storage had no effect on off-flavor in products containing SA. Juiciness was not affected by blend or storage. Results indicate acceptable patties can be made from blends of pork and turkey containing relatively high levels of turkey.     

水分含量对淡腌大黄鱼贮藏性的影响

以感官、微生物和化学变化为指标,探讨了水分含量对淡腌大黄鱼贮藏性的影响。结果表明,水分含量为60%±1%、55%±1%、50%±2%的样品货架期分别是5、9、11d。3组样品货架期终点时菌落总数(TVC)值约为8lg(cfu/g),总挥发性盐基氮(TVB-N)值约为30mg/100g,硫代巴比妥酸反应物(TBARS)值在贮藏期间均出现峰值。贮藏初期菌相比较单一,共检测到4种细菌,贮藏过程中菌相发生变化,货架期终点时仅残存2种细菌,分别为沃氏葡萄球菌(Staphylococcus warneri)和洛菲不动杆菌(Acinetobacter lwoffii)。     

Numbers and types of microorganisms in vacuum-packed cold-smoked freshwater fish at the retail level

Fifty-four packages (each one belonging to a different lot) of vacuum-packed cold-smoked salmon (30) and trout (24) produced by six Spanish smokehouses were obtained at retail level after 3 weeks storage at 2+/-1 degrees C. Sensorial, chemical, physicochemical and microbiological characteristics were examined. Overall, pH, a(w), salt content in water phase, aerobic plate counts at 30 and 25 degrees C. levels of Enterobacteriaceae, lactic acid bacteria (LAB), fungi and presumptive aeromonads and staphylococci are in agreement with available data on lightly preserved fish products. Psychrotrophic clostridia ranged between 1.71 and 2.21 log CFU/g. Levels of ethanol were highly variable and not significantly related (p > 0.05) to sensory scores or to microbial numbers. Salmonella, Escherichia coli and Listeria monocytogenes were not detected in any sample. Listeriae other than L. monocytogenes were isolated from three packages. Levels of Staphylococcus aureus lower than 4 log CFU/g were also found in three packages. Among 377 bacteria randomly isolated from aerobic 25 degrees C plate counts, LAB predominated, with Carnobacterium (C. piscicola) and Lactobacillus (eight species) being the genera most frequently found. The second and third major groups were Enterobacteriaceae and Micrococcaceae, respectively. Proteus vulgaris, P. mirabilis and Serratia liquefaciens were dominant among Enterobacteriaceae and coagulase-negative staphylococci among Micrococcaceae. Minor microbial groups such as aerobic gram-negative bacilli (Acinetobacter; Moraxella and Pseudomonas), Brochothrix, Aeromonas, Bacillus and Vibrio constituted less than 17% of the total flora.     

Validation of time and temperature values as critical limits for Salmonella and background flora growth during the production of fresh ground and boneless pork products

To provide pork processors with valuable data to validate the critical limits set for temperature during pork fabrication and grinding, a study was conducted to determine the growth of Salmonella serotypes and background flora at various temperatures. Growth of Salmonella Typhimurium and Salmonella Enteritidis and of background flora was monitored in ground pork and boneless pork chops held at various temperatures to determine growth patterns. Case-ready modified atmosphere packaged ground pork and fresh whole pork loins were obtained locally. Boneless chops and ground pork were inoculated with a cocktail mixture of streptomycin-resistant Salmonella to facilitate recovery in the presence of background flora. Samples were held at 4.4, 7.2C, and 10 degrees C and at room temperature (22.2 to 23.3 degrees C) to mimic typical processing and holding temperatures observed in pork processing environments. Salmonella counts were determined at regular intervals over 12 and 72 h for both room and refrigeration temperatures. No significant growth of Salmonella (P < 0.05) was observed in boneless pork chops held at refrigeration temperatures. However, Salmonella in boneless pork chops held at room temperature had grown significantly by 8 h. Salmonella grew at faster rates in ground pork. Significant growth was observed at 6, 24. and 72 h when samples were held at room temperature, 10 degrees C, and 7.2 degrees C, respectively. No significant growth was observed at 4.4 degrees C. Background flora in ground pork samples increased significantly after 10 h at room temperature and after 12 h for samples held at 10 and 7.2 degrees C. Background flora in samples held at refrigeration temperatures did not increase until 72 h. Background flora in the boneless chops increased significantly after 6 h at room temperature and after 24 h when held at 10 and 4.4 degrees C. These results illustrate that meat processors can utilize a variety of time and temperature combinations as critical limits to minimize Salmonella growth during production and storage of raw pork products.     

冷却猪肉初始菌相分析与冷藏过程中的菌相变化规律研究

本文采用选择性培养基对托盘包装冷却猪肉的初始菌相进行了分析,并研究了在0、2、5和7℃储藏温度下的菌相变化规律。结果表明:假单胞菌、热死环丝菌、肠杆菌科和乳酸菌是构成有氧条件下冷却猪肉初始菌相的主要微生物。在不同的储藏温度下,细菌总数均有不同程度的增长。假单胞菌在所研究的温度下均与细菌总数相当,增长速度最快。热死环丝菌和肠杆菌也有不同程度的增长,在实验时间的末期也达到了较高的对数值。     

SODIUM ACID PYROPHOSPHATE IN LINKED SAUSAGES MADE WITH PORK AND/OR GOAT MEAT

Five combinations of pork and goat were used to formulate linked sausages in reciprocal increments of 25% (e.g., 100% pork, 75% pork/25% goat meat, etc.). Each formulation was made with or without 0.25% sodium acid pyrophosphate (SAPP), stored in a retail case at 3°C and sampled at 0, 3, 6, 9, and 12 days. Sausages made with 25 or 50% goat meat, regardless of SAPP, were not different (P> 0.05) in off flavor from sausages made with 100% pork and SAPP. SAPP decreased pH values, reduced percentages of cooking loss in 4 of 5 comparisons and slightly improved visual color scores for sausages during initial storage periods of 0 and 3 days.     

冷却猪肉新鲜度的色差快速分析评价方法

为了得到冷却猪肉新鲜度的变化规律,本研究以冷却猪肉里脊部分为研究对象,进行球蛋白沉淀实验、感官评价、pH值、水分含量、肉浸液以及肌肉表面颜色值测定。结果表明：球蛋白沉淀实验、pH值能够较好地反应肉品新鲜度的变化,色差分析符合一定的规律：肌肉表面色差测定中,20℃条件下,a*变化与贮藏时间显著相关(P＜0.05),L*变化显著(P＜0.05);4℃条件下,b*、C变化与贮藏时间极显著相关(P＜0.01),L*值与pH值极显著相关(P＜0.01);对于肉浸液,L*、b*、C、ΔE*均发生显著性变化(P＜0.05),20℃条件下L*、b*、C、ΔE*与贮藏时间和pH值显著相关(P＜0.05),而4℃条件下,L*、C、ΔE*与贮藏时间显著相关(P＜0.05),L*、a*、b*、C、ΔE*值与pH值极显著相关(P＜0.01);冷链过程中,当L*高于48.73、a*高于4.72、b*高于9.18时可认定冷却肉为新鲜肉。     

Survival of Campylobacter jejuni and Salmonella enterica Typhimurium in vacuum-packed, moisture-enhanced pork

The abilities of Campylobacter jejuni and Salmonella enterica Typhimurium to survive in vacuum-packaged, moisture-enhanced pork stored at 4 or 10°C were examined. Pork loins were surface inoculated with either C. jejuni or Salmonella Typhimurium and then moisture enhanced to a target of 10 or 20%. The enhanced pork loins were sliced 1 cm thick and vacuum packaged. A pork loin without moisture enhancement was sliced and vacuum packaged as a control. Samples were collected, plated, and the numbers of surviving organisms were determined periodically during storage at 4 and 10°C. The numbers of C. jejuni or Salmonella Typhimurium in samples with different moisture enhancement levels were similar (P > 0.05). No significant differences (P > 0.05) in C. jejuni counts were observed between samples at 10°C and those at 4°C. In contrast, the numbers of Salmonella Typhimurium in samples at 10°C had significantly (P < 0.05) increased (0.41 log CFU/g) from those at the refrigerated temperature of 4°C. Vacuum storage at 4 and 10°C for 28 days did not result in dramatic reductions in the mean numbers of C. jejuni and Salmonella Typhimurium. Our findings indicate that vacuum packaging under chilled conditions will not add substantially to safety for moisture-enhanced pork. Strict hygienic practices or the implementation of decontamination technologies is recommended.     

Effect of different concentrations of carbon dioxide and oxygen on the growth of pathogenic Yersinia enterocolitica 4/O:3 in ground pork packaged under modified atmospheres

The influence on Yersinia enterocolitica counts of a gradual increase of carbon dioxide concentrations (percentage by volume in air) during packaging and storage of ground pork meat artificially contaminated with this pathogen was evaluated. Ground meat was packaged under customary conditions using modified atmospheres with various carbon dioxide percentages (0, 30, 50, 70, and 100% CO2 by volume; for atmospheres of less than 100% CO2, the rest of the gas was O2). The packs were stored at 2 degrees C for 12 days. During the entire storage time, counts of Y. enterocolitica were determined by the spread plate method for direct plate counts (DPCs). Microbiological shelf life of the stored ground pork also was assessed by total mesophilic aerobic bacterial plate counts (APCs). Y. enterocolitica counts were not significantly different (P > or = 0.05) in the ground pork packaged under the various CO2-enriched atmospheres. The growth of Y. enterocolitica was nearly entirely inhibited in all tested modified atmospheres containing the protective CO2. However, in ground pork packaged with 100% oxygen, there was a significant decrease (P < or = 0.05) in the DPC for Y. enterocolitica from 4.30 log CFU/g (day 0) to 3.09 log CFU/g at the end of the storage time (day 12). The decrease was presumably due to the marked increase in APC seen only in those packages stored under 100% O2. Packaging with high CO2 concentrations had significant inhibitory effect (P < or = 0.05) on the growth of mesophilic aerobic bacteria.     

北方储藏稻谷真菌多样性分析

本研究选择北方有代表性的地区,以高大平房仓常规储存的稻谷为研究对象,根据仓内储藏环境的差异进行分点分层取样。采用传统方法进行真菌分离纯化,通过形态学和分子系统学相结合进行菌株的分类鉴定,通过研究真菌分布为稻谷储藏过程监控提供一定的指导。结果表明,我国北方地区稻谷储藏期含水量为13.6%～15.6%,每个省储藏稻谷真菌菌落总数范围在10~3～10~4 CFU/g之间;北方储藏稻谷共分离出58种真菌,分布于2门,4纲,6目,13科,16属,主要优势菌阿姆斯特丹曲霉Aspergillus amstelodami和多育曲霉Aspergillus proliferans被划分为灰绿曲霉群及其近缘类群,这类真菌对于仓储环境的适应性较强,可作为北方粮堆异常粮情早期监测预警的主要指示菌群;粮堆上层或靠近墙壁等易受环境温度影响的位置,易达到储藏真菌的生长条件,优势菌群由初始的田间和过渡真菌演替为储藏真菌,是高大平房仓稻谷储存过程中需要重点关注的位置。确定储藏稻谷的优势菌群及仓房重点关注位置,为北方稻谷储藏过程提供了技术参考。     

冷链运输中不同包装方式对生鲜猪肉品质的比较分析

为了使生鲜肉类能够在冷链运输中保持良好的品质,分别研究了在冷链运输中三种不同包装方式下对生鲜猪肉品质的影响。本实验选取生鲜猪肉为研究对象,通过不同方法分别测定了不同包装方式下生鲜猪肉TVB-N含量、不同包装方式下生鲜猪肉p H值、不同包装环境下菌群数量的变化以及不同包装方式下生鲜猪肉的脂肪氧化程度(TAB)等。通过对这些参数的分析,得到不同包装方式冷链运输中生鲜猪肉品质的具体数据。通过实验分析可知,气调包装方式下生鲜猪肉TVB-N含量均在15mg/100g以下,三种不同包装方式下生鲜猪肉的PH值均在5~7之间,均在合理范围内,气调包装在生鲜猪肉冷链运输中可有效抑制菌群数量变化,菌群数量控制在10~3 CFU/g^10~4 CFU/g之间,而且在冷链运输过程中生鲜猪肉颜色等保持较好;真空包装方式下TBA的最高值为0.45mg/kg,能够效抑制肉类的脂肪氧化。     

The bacteriology of fresh and spoiling Lake Victorian Nile perch (Lates niloticus)

A total of 177 bacterial cultures isolated from Lake Victorian Nile Perch (Lates niloticus) were investigated. The flora on newly caught Nile perch consisted of organisms belonging to the genera Moraxella, Alcaligenes, Acinetobacter, Pseudomonas, Aeromonas, Micrococcus and other Gram-positive organisms. 39% were identified as Gram-positive species and 61% were negative in the Gram-reaction. Three cultures out of 53 investigated caused weak rotten off-odours in sterile fish broth and one culture, an Aeromonas spp. produced strong rotten, fishy, hydrogen sulphide off-odours. From Nile perch spoiled at ambient temperature, 15 of the 42 strains isolated caused rotten, fishy, hydrogen sulphide off-odours. These specific spoilage bacteria were all identified as Aeromonas and all reduced trimethylamine oxide to trimethylamine and produced hydrogen sulphide. From spoiled iced Nile perch, 74 out of 82 (90%) of the bacteria isolated were identified as Pseudomonas. A small proportion of these (13 out of 74) produced off-odours in sterile fish broth resembling the spoiling fish. These specific spoilers could not be separated from the non-spoilers based on biochemical activities used in classical taxonomy. While the Pseudomonas spp. isolated did not produce trimethylamine or H₂S, a few of the remaining isolates (two Shewanella putrefaciens and five Aeromonas spp.) did produce these compounds. The role of Shewanella putrefaciens in the iced spoilage of Nile perch was, however, insignificant, since they only very late in the storage reached numbers where their spoilage could be detected.     

基于Illumina MiSeq测序技术分析大鲵肉冷藏过程中微生物菌群演替规律

为探究大鲵肉冷藏过程中菌相组成及特定腐败微生物,对托盘包装大鲵肉不同冷藏时间（4 ℃,0、2、4、6、8 d）的挥发性盐基氮和菌落总数进行评价,在此基础上采用Illumina MiSeq测序技术探究其微生物菌群变化与多样性。结果表明,大鲵肉在冷藏过程中菌落总数和挥发性盐基氮呈上升趋势,分别在第6天和第8天超标。高通量测序结果显示,大鲵肉中微生物丰度随着冷藏时间的延长呈降低趋势。在门水平上进行群落组成分析,细菌的优势菌门从冷藏前期（0、2 d）的拟杆菌门、厚壁菌门逐渐转变为中（4 d）、后期（6、8 d）的变形菌门。在属水平上细菌的优势菌属从冷藏前期主要的拟杆菌属、Faecalibacterium逐渐转变为中、后期的假单胞菌属、气单胞菌属、Hafnia-Obesumbacterium、沙雷氏菌属。主坐标分析显示0-2、4、6、8 d之间微生物菌群差异较大,2 个主坐标叠加解释度达80.92%;LEfSe分析表明,引起大鲵肉各冷藏期差异显著的菌门主要为变形菌门、放线菌门、拟杆菌门、纤维杆菌门、厚壁菌门,菌属主要为假黄单胞菌属、Bauldia、沙雷氏菌属、不动杆菌属、气单胞菌属、假单胞菌属、ambiguous_taxa、Hafnia-Obesumbacterium、Rikenellaceae_RC9_gut_group、Prevotella_9、拟杆菌属、纤维杆菌属、毛螺菌属、Faecalibacterium、Clostridium_sensu_stricto_1。进化分析表明大鲵肉冷藏过程中微生物菌群演替与冷藏时间具有较强相关性。综合分析,引起冷藏期间大鲵肉腐败变质的微生物主要为假单胞菌属、气单胞菌属、Hafnia-Obesumbacterium和沙雷氏菌属。该研究为今后大鲵肉冷藏过程中靶向抑菌及货架期延长提供了参考。     

Microbial profiles of commercial, vacuum-packaged, fresh pork of normal or short storage life

The microbial ecology of fresh vacuum-packed pork cuts during storage at -1.5 degrees C for up to 45 days was examined to characterize rates of microbial growth and pH changes in commercially prepared products of normal storage quality. Pork loins in commercial distribution with odour defects were also studied to determine a possible cause of the defects and avoid future problems. In addition, microbial profiles of pork cuts from two plants were compared, after storage for 25 days at -1.5 degrees C, to identify possible reasons for differences in the storage life of product from the plants. The effects of a change in sanitation procedures on the microbial populations of products stored for 25 days were also studied. With normal product, microbial growth in different packages progressed at different rates, reflecting differences in initial levels of bacterial contamination. All samples in the study reached 8 weeks without apparent organoleptic change and samples carried 5.8+/-1.2 log bacteria cm(-2) (mean+/-S.D.). The flora of loins with the odour defect were predominately lactic acid bacteria (LAB) and carnobacteria, but they contained large fractions of Enterobacteriaceae <35 days after packaging. Aeromonas spp. and Shewanella spp. were likely responsible for the sulfide-putrid smell of these spoiled products, but species of Enterobacteriaceae and lactic acid bacteria could have contributed to spoilage. Comparison of microbial groups present in 16 other cuts, half from each of two commercial plants, which were stored for 25 days at -1.5 degrees C, showed that larger fractions of Enterobacteriaceae were present in samples from the plant having difficulty achieving the desired storage life. Additional bacterial samples from 12 cuts supplied by the latter plant obtained after adoption of an acid sanitizer step in the plant cleaning regimen, and also stored for 25 days at -1.5 degrees C, yielded few Enterobacteriaceae, Aeromonas or Shewanella. Use of an acid sanitizer in plant cleaning may be a means of controlling alkali-tolerant bacteria such as Aeromonas or Shewanella which can contaminate pork cuts and spoil vacuum-packaged product. The fraction of Enterobacteriaceae in bacteria populations on fresh pork stored for 25 days at -1.5 degrees C may be a useful indicator of the effectiveness of plant sanitation.     

冷却猪肉不同前处理对细菌DNA提取及PCR-DGGE的影响

比较研究冷却猪肉不同前处理对细菌DNA 提取及PCR- 变性梯度凝胶电泳(DGGE)的影响。冷却猪肉4℃贮藏至第5 天时,分别采用冻融、超声、摇床和匀浆前处理后,提取细菌DNA,进行Dilution-PCR 和DGGE 分析。结果表明,不同前处理方法所制备的DNA 量稍有差别,但对PCR-DGGE 结果无显著影响,故可根据实际情况选择上述不同前处理方法。同时对16S rDNA V3 区的DGGE 图谱进行割胶测序,检测到腐败菌主要是假单胞菌属,其他还有不动杆菌属、环丝菌属和沙雷氏菌属。     

低钠盐火腿肠保质期预测及产品致病菌的聚合酶链式反应检测

采用加速破坏性试验模型预测低钠盐火腿肠的保质期,并采用聚合酶链式反应对产品致病菌进行快速检测,同时对贮藏期间火腿肠的质构(嫩度)变化进行分析。结果表明：对照组(3%普通碘盐＋0.3%磷酸盐)、2.5%市售低钠盐＋0.35% TG组、2.0%市售低钠盐＋0.3%磷酸盐组、3%多组分替代盐(NaCl 40%、KCl 35%、CaCl2 15% 和甘氨酸10%)＋0.35% TG组火腿肠在室温(25℃)和低温(4℃)条件下的保质期分别为17.5、17.6、16.9、15.7d和189、208、142、146d;产品在低温(4℃)条件下放置1个月后,各组均无致病菌检出;火腿肠在低温贮藏90d期间,其嫩度变化趋势是先增大后减小。综合比较而言,添加2.5%市售低钠盐+0.35% TG组火腿肠的贮藏稳定性最好。     

Identification of Microbial Isolates from Vacuum-Packaged Ground Pork Irradiated at 1 kGy

Bacterial cultures from irradiated (1 kGy) and nonirradiated, vacuum-packaged ground pork held at 5°C were isolated and characterized over a 12-day storage period. The initial flora of the meat was composed mostly of Pseudomonas sp. and Enterobacter sp. Although the microflora of nonirradiated samples gradually shifted from Gram-negative to Gram-positive microorganisms, 76% of the isolates were characterized as Gram-negative at the onset of spoilage (9 days at 5°C). In contrast, the irradiated ground pork microflora was mainly Gram-positive (66%) shortly after irradiation and increased to 97% after 9 days at 5°C. A total of 720 isolates were identified to genus.     

猪肉新鲜度指示标签的制备及应用

基于猪肉在腐败过程中产生的挥发性含氮化合物会使包装中气体酸碱性发生变化的原理,以甲基纤维素和聚乙二醇-6000为基材,添加溴甲酚紫、溴百里酚蓝等酸碱指示剂,最终制得6 种方便、无损的新鲜度指示标签。将标签置于不同质量浓度的NH3环境中,6 种标签的颜色均发生显著变化,且随NH3质量浓度的增加而差异变化明显;将标签应用于生鲜猪肉贮存中,通过测定4 ℃条件下不同贮存时间猪肉各项腐败指标以及指示标签的颜色变化,探究其用于监测生鲜猪肉新鲜度的可行性。结果表明：初始pH值为4.24的溴甲酚紫指示标签可较好地区分出猪肉的3 个新鲜度等级;利用此标签的颜色测定数据建立偏最小二乘模型,对挥发性盐基氮含量和菌落总数进行预测的回归系数（R2）均大于0.92,通过标签颜色变化可以判断猪肉新鲜度。     

基于Illumina MiSeq高通量测序分析清香型白酒酒糟微生物群落多样性

采用Illumina MiSeq高通量测序技术对清香型白酒酒糟中微生物多样性进行研究。结果表明,酒糟样品中细菌菌群的丰富度与多样性均大于真菌,细菌菌群归属于10个门、18个纲、31个目、66个科、86个属、124个种,真菌菌群归属于5个门、11个纲、20个目、33个科、49个属、66个种。优势细菌门（相对丰度≥1.0%）为变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidetes）,优势细菌属为根瘤菌属（Rhizobium）、拉乌尔菌属（Raoultella）、谷氨酸杆菌属（Glutamicibacter）、红球菌属（Rhodococcus）、短波单胞菌属（Brevundimonas）、不动杆菌属（Acinetobacter）、假单孢菌属（Pseudomonas）、嗜冷杆菌属（Psychrobacter）、Devosia、嗜盐单胞菌属（Halomonas）、未分类-鼠杆菌属（unclassified-Muribaculaceae）、乳杆菌属（Lactobacillus）;优势真菌门为子囊菌门（Ascomycota）、毛霉门（Mucoromycota）及担子菌门（Basidiomycota）,优势真菌属为孢圆酵母属（Torulaspora）、伊萨酵母属（Issatchenkia）、横梗霉属（Lichtheimia）、根霉属（Rhizopus）、曲霉属（Aspergillus）、酵母属（Saccharomyces）。