包装冷却肉中微生物腐败及其挥发性气味的研究进展

肉的腐败气味一直是消费者评判肉品新鲜度的重要指标。不同包装冷却肉中的微生物群落和代谢途径不同,导致肉品贮藏期间释放出不同的挥发性气味物质,这直接决定了肉的腐败气味和腐败程度;因此,进行包装肉挥发性物质的定性、定量分析对认知肉品腐败进程具有重要意义。本文通过综述冷却牛肉、猪肉、羊肉及禽肉在托盘包装、真空包装、气调包装中的微生物群落演替及其典型挥发性物质变化过程,揭示了特定包装过程中诱发肉品腐败气味的优势菌群,从底物利用和代谢通路方面分别探讨了这些气味的产生机制,并对现有检测手段进行总结,以期为控制肉品腐败异味,利用挥发性标记物监测包装肉品货架期提供思路。     

Spoilage-related microbiota associated with chilled beef stored in air or vacuum pack

In order to study the spoilage-related microbiota of beef at species level, a combination of culture-independent and culture-dependent methods was used to analyse nine different beef samples stored at 4 °C in air or in vacuum pack. Plate counts on selective agars after 0, 7 and 20 days of storage showed that vacuum packaging reduced the viable counts of Brochothrix thermosphacta, Pseudomonas spp. and Enterobacteriaceae, whereas the growth of lactic acid bacteria (LAB) was unaffected. Storage in vacuum pack mainly affected viable counts and not necessarily the species diversity of microbial populations on meat. Such populations were studied by PCR-DGGE of DNA directly extracted from meat and from bulk cells from culture media, followed by sequencing of DGGE fragments. Pseudomonas spp., Carnobacterium divergens, B. thermosphacta, Rahnella spp. and Serratia grimesii, or close relatives were detected in the meat at time zero. The use of the culture-independent method highlighted the occurrence of species that were not detected by plating. Photobacterium spp. occurred in most meat samples stored in air or in vacuum pack, which indicates this organism probably has a role in spoilage. In contrast, culture-dependent analysis allowed detection of bacterial species that were not found in DNA extracted directly from meat. This was the case for several species of Serratia or Rhanella among the enterobacteria, and Leuconostoc spp. among the LAB. Besides advancing our knowledge of the species involved in the spoilage of vacuum-packaged meat, this study shows the benefits of combining culture-based and direct approaches to enhance understanding of populations of spoilage bacteria.     

生物保鲜剂延长冷鲜牛肉货架期的效果

为延长冷鲜牛肉的货架期,保证其贮藏期间的食用品质及安全性,采用单因素试验与正交试验筛选一组 复配生物保鲜剂（乳酸链球菌素、纳他霉素和ε-聚赖氨酸）,并研究其对冷鲜牛肉的保鲜效果。结果表明：复配 生物保鲜剂中3 种保鲜剂的最佳配比为乳酸链球菌素2.5 g/L、纳他霉素1.5 g/L、ε-聚赖氨酸3.0 g/L;冷鲜牛肉在 （4±1） ℃条件下贮藏8 d后,其挥发性盐基氮值为19.84 mgN/100 g,属于二级鲜肉;菌落总数较对照组降低2 个 数量级,表明复配生物保鲜剂能够有效抑制腐败菌的增长;贮藏后期,处理组冷鲜牛肉的汁液流失率较对照组减少 1.73%。复配生物保鲜剂可以将冷鲜牛肉的货架期有效延长4 d。     

Role of biogenic amines as index of freshness in beef meat packed with different biopolymeric materials

The main objectives of this work were to evaluate the chemical and microbiological fresh beef meat quality packed in aerobic atmosphere with biopolymers, to investigate the possible role of biogenic amines (BAs) as indicators of spoilage in fresh beef meat stored at 4 °C for 8 days. The results of this research highlighted that for fresh meat packaging it could be possible to replace the PS tray/PVC film system, with an expanded PLA biopolymeric tray heat-sealed with a biopolymeric film, characterized by a negligible environmental impact in comparison with the use of synthetic plastic materials. The storage time differentiated the meat samples on the basis of pH and microbiological characteristics. With regard to BAs, tyramine and cadaverine resulted strongly influenced by the storage time, and to a less extent putrescine and spermidine. Tyramine and cadaverine could be used as spoilage indexes of fresh beef meat chilled and packed in aerobic atmosphere with biopolymers.     

The effective product life of vacuum-packaged beef imported into Saudi Arabia by sea, as assessed by chemical, microbiological and organoleptic criteria

Five transport and storage trials were conducted on commercial shipments of vacuum-packaged beef half striploins imported into Saudi Arabia by sea from Australia, New Zealand and the Republic of Ireland. The cold chains, from packaging to the end of chilled storage in Saudi Arabia, were monitored using miniature electronic data loggers placed in cartons of striploins. Development of spoilage microfloras was followed from packaging to trial end using a differential aerobic plate count technique. Changes in meat pH, total volatile nitrogen (TVN) and free fatty acid (FFA) content were determined over the storage period. Sensory evaluations to identify spoilage conditions and to determine consumer acceptability were also conducted over the course of storage in Saudi Arabia. Results indicated that aerobic plate counts, TVN and FFA were all unsuitable as indicators of the fitness, or acceptability of vacuum-packaged beef for human consumption. Under good but commercially realistic conditions (i.e. normal ultimate pH beef, initial microbial contamination at packaging of less than 10³/cm², packaging, film oxygen transmission rate less than 40 ml/m²/24 h/atm at 23°C and 90% R.H., and mean product temperature during transport and storage of 0°C), vacuum-packaged beef transported by sea has an anticipated product life of at least 90 days measured from the date of slaughter. Under the conditions just stated, deterioration of meat texture resulting from excessive aging rather than the onset of overt microbial spoilage will limit the effective product life of vacuum-packaged beef.     

Use of nisin, lysozyme and EDTA for inhibiting microbial growth in chilled buffalo meat

The effect of nisin, lysozyme and lysozyme combined with disodium ethylenediaminetetraacetate salt (EDTA) for inhibiting the growth of spoilage micro‐organisms in buffalo meat was investigated. Prior to packaging, the samples were alternatively dipped in solutions containing different concentrations of nisin, lysozyme and combinations of a pre‐fixed amount of lysozyme and different concentrations of EDTA. The packed meat samples were stored at 4 °C, and the microbial changes were monitored for a period of 8 days. The best results, in terms of microbial shelf‐life extension, were obtained by dipping the meat steaks in a solution containing a combination of 0.5% of lysozyme and 2% of EDTA. This treatment was able to reduce the growth of all the investigated bacteria and showed a bactericidal effect on Brochotrix thermosphacta during the entire period of storage.     

冷鲜牛肉贮藏中菌群结构及优势菌致腐性的分析

为探究牛肉在0?℃冷藏下微生物菌群变化和优势腐败菌,采用培养依赖的16S rRNA结合高通量测序技术分析牛肉样品的微生物多样性变化。通过定期测定接种牛肉汁的生长、感官、蛋白酶活性、pH值和挥发性盐基氮（total volatile basic nitrogen,TVB-N）值,分析分离株的致腐特征。结果表明,牛肉冷藏中感官品质保持良好,15?d出现异味,18?d有腐臭味。而样品中菌落总数第3天快速上升,15?d菌落总数为8.73（lg（CFU/g））,之后呈现稳定,假单胞菌（Pseudomonas spp.）、热死环丝菌（Brochothrix thermosphacta）、肠杆菌（Enterobacter）和乳酸菌4 种分离培养基中细菌与菌落总数增长趋势相似,其中假单胞菌增长最快,肠杆菌数和乳酸菌数生长最慢。两种菌群鉴定结果显示,冷鲜牛肉初始微生物构成复杂,包括热死环丝菌、不动杆菌属（Acinetobacter spp.）、气单胞菌属（Aeromonas spp.）和假单胞菌等多种菌属构成,而腐败末期菌群构成趋于单一,假单胞菌和热死环丝菌为优势腐败菌,特别是莓实假单胞菌（P. fragi）。将腐败分离株10?株假单胞菌、4?株热死环丝菌和1?株蜂房哈夫尼菌（Hafnia alve）接种于冷藏的牛肉汁,发现假单胞菌和蜂房哈夫尼菌的接种组感官评分、pH值和TVB-N值高于热死环丝菌,且假单胞菌有较强的蛋白酶活性。研究表明,结合感官和微生物评价冷鲜牛肉贮藏期为15?d,且菌群多样性下降,假单胞菌和热死环丝菌是优势腐败菌,其中假单胞菌致腐性较强。     

冷藏卵形鲳优势腐败菌的分离鉴定及致腐能力分析

为研究卵形鲳鲹冷藏末期的优势腐败菌及其致腐能力,采用传统选择性培养结合16S rDNA序列分析法,对卵形鲳鲹4℃贮藏期间腐败菌分别进行分离纯化、菌相分析及菌属确定,并以腐败物质的产量因子Y_TVB-N/CFU为评价标准,定量分析了贮藏末期优势菌的致腐能力。结果得出,卵形鲳鲹4℃贮藏过程中共分离纯化出16株细菌,其中革兰氏阴性菌10株,革兰氏阳性菌6株,其分属6个菌属。贮藏初期（0 d）时菌相丰富,希瓦氏菌属（Shewanella sp.）、假单胞菌属（Pseudomonas sp.）、葡萄球菌属（Staphylococcus sp.）、芽孢杆菌属（Bacillus sp.）、乳酸杆菌属（Lactobacillus sp.）与肠杆菌属（Enterobacter sp.）等均有出现;中期（3 d）菌群种类有所减少,以希瓦氏菌属（Shewanella sp.）与假单胞菌属（Pseudomonas sp.）所占比例较高,后期（6 d）种类进一步减少,只有3株菌占较高,分别是2号腐败希瓦氏菌（Shewanella putrefaciens 48%）、5号奥奈达希瓦氏菌（Shewanella oneidensis 26.6%）与16号霍氏肠杆菌（Enterobacter hormaechei 19.4%）;致腐能力由高至低依次为腐败希瓦氏菌 > 奥奈达希瓦氏菌 > 霍氏肠杆菌,其中腐败希瓦式菌致腐能力显著强于另两株菌;综合贮藏末期所占比例及致腐能力结果,最终确定腐败希瓦氏菌为4℃贮藏下卵形鲳鲹的优势腐败菌。     

Spoilage microbiota associated to the storage of raw meat in different conditions

The spoilage of raw meat is mainly due to undesired microbial development in meat during storage. The type of bacteria and their loads depend on the initial meat contamination and on the specific storage conditions that can influence the development of different spoilage-related microbial populations thus affecting the type and rate of the spoilage process. This review focuses on the composition of raw meat spoilage microbiota and the influence of storage conditions such as temperature, packaging atmosphere and use of different preservatives on the bacterial diversity developing in raw meat. In addition, the most recent tools used for the detection and identification of meat microbiota are also reviewed.     

冷鲜鸡胸肉主要腐败菌的分离及低温贮藏对货架期的影响

冷鲜鸡肉的货架期与其腐败菌的种类和数量密切相关。采用选择性培养基对冷鲜鸡胸肉中的主要腐败菌进行了分离,并对其低温条件下的货架期进行了研究。结果表明,假单胞菌、热死环丝菌、肠杆菌和乳酸菌是冷鲜鸡胸肉中的主要腐败菌;在低温贮藏过程中,前两种菌的数量呈明显上升趋势,并成为优势菌群。在2、4、8和10℃贮藏条件下,冷鲜鸡胸肉的货架期分别为14、10、5和3 d。     

A combination of modified atmosphere and antimicrobial packaging to extend the shelf-life of beefsteaks stored at chill temperature

An antimicrobial polyethylene (PE) film was obtained by coating a nisin-based antimicrobial solution. PE sheets were coated on both sides and were used for the packaging of beefsteaks to be stored in air or modified atmosphere packaging (MAP, 60% O(2)-40% CO(2)). Microbial populations, species diversity, headspace volatile organic compounds, colour and sensory properties were monitored after 0, 1, 7 and 12days of storage at 4°C. The viable counts showed that there was an effect of MAP and antimicrobial film on the development of all the spoilage associated microbial populations. Carnobacterium spp., Brochothrix thermosphacta, Pseudomonas fragi and Rhanella aquatilis were found in most of the samples. C. maltaromaticum was identified in MRS bulk cells from samples stored in air as well as MAP. Quantitative data of headspace-SPME-GC/MS analysis showed that during storage the production of volatile organic compounds (VOCs) was affected by the use of the treated film and the MAP storage. Compounds such as phenylethylalcohol, nonanal, decanal and ethylbutanoate were produced only from 7 to 12day of storage and only in the samples stored in air. In agreement with the microbiological and VOCs data, the meat stored in active packaging scored the best rankings in the sensory evaluation. Principal component analysis of microbial, sensory and instrumental data showed that beefsteaks stored with the combination of MAP and active packaging for 12days at 4°C differed from the other samples that were more associated to high microbial loads, VOCs concentration and meat off odour perception. In conclusion, the antimicrobial sheets in combination with MAP storage at 4°C were effective for the storage of beefsteaks by retarding the growth of spoilage bacteria, determining lower concentration of VOCs and keeping acceptable levels of colour and other sensory parameters for more than 10days.     

Comparison of culture-dependent and independent techniques for characterisation of the microflora of peroxyacetic acid treated,vacuum-packaged beef

The diversity of microflora associated with peroxyacetic acid (POAA) treated and untreated beef was investigated by 16S rDNA gene cloning, DGGE analysis and conventional bacterial cultivation. Following vacuum packaging, POAA treated and untreated meat samples were stored for up to 18 weeks at −1.5 °C. Each culture independent method showed Carnobacterium spp. to predominate on both POAA treated and untreated meat. However, 16S rDNA gene analysis also detected the presence of psychrotolerant Clostridium spp. in the POAA-treated beef. Culture-dependent analysis did not distinguish Carnobacterium spp. from Lactobacilli. Although culture-dependent analysis showed an increase in the ratio of Enterobacteriaceae to lactic acid bacteria from weeks 6–18 in the POAA treated compared with the untreated meat, the numbers of Enterobacteriaceae were significantly less on POAA treated than on untreated meat. The combination of data collected by culture-dependent and independent techniques provided the most robust approach for elucidating the efficacy of chemical sanitization of chilled vacuum-packaged beef. If conventional cultivation is used for monitoring bacterial spoilage of vacuum-packaged chilled meats it is recommended that culture methods specific for Carnobacterium and Clostridium spp. should be included in order to provide a more complete indication of microbial diversity.     

Reduction of the spoilage-related microflora in absorbent pads by silver nanotechnology during modified atmosphere packaging of beef meat

Silver-based antibacterial hybrid materials have been developed by in situ reduction of silver nitrate (1%) adsorbed on cellulose fibers by thermal and UV treatments. Microscopy revealed that the silver nanoparticles were dispersed and regular in shape. Migrated silver ions achieved 60 ppm in beef meat exudates. The ability of the silver-loaded absorbent pads to lower microbial contamination of exuded fluids was studied during storage of beef meat in modified atmosphere packaging. Cellulose-silver hybrid materials reduced the levels of the major microbial groups (total aerobic bacteria, lactic acid bacteria, Pseudomonas spp., and Enterobacteriaceae) present in the absorbent pads by an average of 1 log CFU/g during the entire storage period. The levels of total aerobic bacteria and Pseudomonas spp. were significantly reduced in the presence of silver ions, whereas lactic acid bacteria were less sensitive and not significantly affected. Enterobacteriaceae levels remained under the detection limit when silver was present. Neither the color of the meat nor the microbial loads were markedly affected by the presence of the silver-based antimicrobial hybrid materials.     

冷却肉微生物腐败与冷链系统

冷却肉极易发生微生物腐败,温度是最关键的影响因素。冷链系统持续的低温能抑制微生物的生长繁殖,预防冷却肉腐败。本文简述了微生物代谢活动对冷却肉腐败的影响,重点将冷链系统分为加工冷链、运输冷链、销售冷链和家庭保藏四部分,分别从概念及存在的问题等方面加以阐述,并展望了其未来有待提高完善的地方。     

Microbial inactivation after high-pressure processing at 600 MPa in commercial meat products over its shelf life

The behaviour of spoilage and pathogenic microorganisms was evaluated after high-pressure treatment (600 MPa 6 min, 31 °C) and during chilled storage at 4 °C for up to 120 days of commercial meat products. The objective was to determine if this pressure treatment is a valid process to reduce the safety risks associated with Salmonella and Listeria monocytogenes, and if it effectively avoids or delays the growth of spoilage microorganisms during the chilled storage time evaluated. The meat products covered by this study were cooked meat products (sliced cooked ham, pH 6.25, a_w 0.978), dry cured meat products (sliced dry cured ham, pH 5.81, a_w 0.890), and raw marinated meats (sliced marinated beef loin, pH 5.88, a_w 0.985). HPP at 600 MPa for 6 min was an efficient method for avoiding the growth of yeasts and Enterobacteriaceae with a potential to produce off-flavours and for delaying the growth of lactic acid bacteria as spoilage microorganisms. HPP reduced the safety risks associated with Salmonella and L. monocytogenes in sliced marinated beef loin.     

Computer-Assisted Identification of Microflora on Hot-Boned and Conventionally Processed Beef: Effect of Moderate and Slow Chilling Rate

Hot-boned (HB) beef was chilled from ca. 32°C to 21°C within 6, 9, or 11.3 hr after fabrication and continuously chilled to 2°C. The counterparts were conventionally processed (CP) and were used as controls. The bacterial counts on HB beef were proportional to the time required to chill HB beef to 21°C. The number of significantly different phena between HB and corresponding CP beef paralleled the chilling rate. The slower chilling rate resulted in more different microbial profiles and encouraged more growth of lactic acid and saccharolytic bacteria than the faster rate. The microflora at the time of packaging subsequently affected the microflora after storage. The initial chilling rate significantly affected the number and the kinds of organisms that developed on the meat surface. Proper chilling of HB beef will provide a microbiologically safe product.     

生鲜牛肉中的腐败微生物概述

生鲜牛肉腐败是一个涉及多种特定微生物相互作用并受多种因素影响的复杂的微生物生态现象。了解牛 肉中微生物的污染源及微生物的多样性对于控制其腐败进程、延长牛肉货架期十分重要。因此,本文对肉牛从 下刀宰杀到包装前所有可能的污染源、污染微生物在种的水平上的分类和多样性、单个菌属及多种微生物作用的 腐败机制等方面对生鲜牛肉中腐败微生物的相关信息进行了系统性分析,初步总结得出活体动物的皮毛及皮毛上 的污染物、去皮用的刀具和剥皮操作工人的手、分割时的工作接触面为生鲜牛肉中微生物的主要污染源;主要 的微生物种类为假单胞菌（Pseudomonas）、肠杆菌（Enterobacteriaceae）、乳酸杆菌及热杀索丝菌（Brochothrix thermosphacta）;不同种类的微生物因利用肉中营养代谢底物的顺序不同而产生不同的腐败现象。期望通过本文的 总结和概述能够为生鲜牛肉腐败微生物的控制提供理论指导。     

Coating-Activation and Antimicrobial Efficacy of Different Polyethylene Films with a Nisin-Based Solution

Antimicrobial packaging can be considered an extremely challenging technology that could have a significant impact on shelf-life extension and food safety of fresh meat and meat products. In this study, different commercial polyethylene films differing in vinyl acetate ethylene, erucamide contents, and oxygen permeability were used for the coating treatment with a nisin-based antimicrobial solution (NS). Detection and measurement of the activity of the NS was determined against different food spoilage bacteria. NS was then spread manually on food contact layer of different plastic films using coating rods providing thickness of 6, 40, 60, and 100 μm. The polyethylene films before and after treatment were analysed by atomic force microscopy (AFM). NS was active against Gram-positive bacteria and the best activity was obtained against Brochothrix thermosphacta. Viable staining and epifluorescence microscopy analysis of indicator strains in contact with activated plastic films showed that the effect of the film on the various indicator strains changed very much on the basis of both type of film and indicator strain. The highest numbers of lysed cells were shown by two polyethylene films that, according to the AFM and roughness parameters analyses, were characterized by significant increase or decrease of roughness after the coating treatment. AFM analysis showed that the homogeneity of the coating was much influenced by the type of plastic films used. In order to test the efficacy in food, portions of beef chuck tender slices were prepared and covered with the antimicrobial plastic films on both sides. After 1 h and 1, 7, and 12 days of storage at 4 °C the meat samples were analyzed by standard plate counting targeting spoilage associated microbial populations. The antimicrobial plastic films after 1 h of contact with the meat caused a significant reduction of lactic acid bacteria and B. thermosphacta. The most effective antimicrobial activity of films was shown against the same populations after 24 h of storage.     

Reduction of Brochothrix thermosphacta and Salmonella Serotype Typhimurium on Vacuum-Packaged Fresh Beef Treated with Nisin and Nisin Combined with EDTA

ABSTRACT: The effect of nisin and nisin/EDTA followed by vacuum packaging on the reduction of Brochothrix thermosphacta and Salmonella typhimurium counts on beef was investigated. Beef cubes, inoculated with either bacteria, were dipped in nisin, or nisin combined with EDTA solutions, vacuum packaged, and stored at 4 °C for up to 25 d. Growth of B. thermosphacta was completely inhibited by nisin and nisin/EDTA by up to 6.76 log₁₀ CFU/ml. However, neither treatment showed significant effects against S. Typhimurium. Thus, nisin or nisin/EDTA treatment, followed by vacuum packaging under refrigerated conditions could increase the shelf life of fresh beef by inhibiting the growth of B. thermosphacta , an important spoilage organism in beef.     

The Predominance of Psychrotrophic Pseudomonads on Aerobically Stored Chilled Red Meat

Microbial spoilage of meat during chilled aerobic storage causes significant financial losses to the industry. Even with modern day preservation techniques, spoilage remains an unsolved problem. Spoilage of meat is a complex process that involves the activity of endogenous enzymes and microorganisms. Psychrotrophic Pseudomonas species are the key microorganisms that cause spoilage in aerobically stored chilled meat. Spoilage pseudomonads are highly robust and able to withstand stressful environmental conditions that would otherwise inhibit the growth of other spoilage organisms. In order to implement efficient control measures, and to minimize spoilage, a thorough understanding of the characteristics of spoilage pseudomonads is essential. This review focuses on the spoilage process and the key metabolic attributes of the main psychrotrophic spoilage Pseudomonas species to explain their predominance on meat over other psychrotrophic bacteria. This review also highlights less studied, but important, characteristics of psychrotrophic pseudomonads such as biofilm formation and quorum sensing in the context of meat spoilage. The importance of the use of model systems that are closely applicable to the food industry is also discussed in detail.