Changes in Microflora of Hot-boned Pork during Storage as Analyzed by PCR-DGGE

Abstract

Abstract: Traditional microbial culture and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) were used to examine microflora changes of hot-boned pork during storage at different temperatures. Total aerobic counts (TAC) were determined at regular time intervals during storage and bacterial DNA extraction was carried out for PCR-DGGE analysis. The storage time required for reaching the minimum spoilage level 7.2(lg(CFU/g)) at 5, 15, 25 ℃ and 30 ℃ were 14 d, 75 h, 19 h and 16 h, respectively. PCR-DGGE demonstrated that different spoilage bacteria dominated at the end of storage at different temperatures, and the dominant spoilage bacteria in hot-boned pork were mainly Macrococcu, Klebsiella, Pseudomonas, Citrobacter, Acinetobacter and Escherichia.

Key words: hot-boned pork, total aerobic counts(TAC), polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), microflora change

CLC Number:

TS251.51

References

[1] FISHCER S G， LERMAN L S. Length independent separation of DNA restriction fragments in two dimensional gel electrophoresis[J].Cell，1979，16(1):191-200.
[2] MUYZER G，WAAL E C，UITTERLINDEN A G. Profiling of complex and microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes Coding for 16SrRNA[J]. Applied of Environment Microbiology，1993，59(3):695-700.
[3] GURTLER V，GARRIE H D，MAYALL B C. Denaturing gradient gel electrophoretic multilocus Sequence typing of staphycoccus aureus isolates [J]Electrophoresis 2002,23:3310-3320.
[4] THEELEN B，SilVESTRI M，GUEHO E，et al. Identification and typing of Malassezia yeasts using amplified fragment length polymorphisms(AFLP)，random amplified Polymorphic DNA(RAPD) and denaturing gradient eleleetrophoresis(DGGE)[J].FLMS Yeast Research，2001，l:79-86.
[5] COCOLIN L， INNOCENTE N，BIASUTTI M. The late blowing in cheese: a new molecular approach based on PCR and DGGE to study the microfloral ecology of the alteration process[J]. International Journal of Food Microbiology，2004，90:83-91.
[6] LEEA J S，HEOA G Y，LEEA J W，et al. Analysis of kimchi microflora using denaturing gradient gel electrophoresis [J]. International Journal of Food Microbiology，2005，in press.
[7] COCOLIN L，MANZONO M，AGGIO D，et al. A novel polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis(DGGE) for the identification of Micrococcaceae strains involved in meat fermentations. Its application to fermented Italian sausage[J].Meat Science，2001，58:59-64.
[8] 许爱清，李宗军，王远亮，等.应用PCR-DGGE技术检测发酵食品和饲料中真菌菌群[J].食品科学，2010，31（7）：317-321.
[9] 周志刚，石鹏君，姚斌，等.基于PCR-DGGE指纹图谱川纹笛鲷及圆白鲳消化道壁优势菌群结构比较分析[J].水生生物学报，2007,31（5）：682-685.
10] 江芸，高峰，徐幸莲，等.真空包装冷却猪肉冷藏过程中菌相变化[J].食品科学，2011,32（4）：241-245.
[11] JIANG Yun, GAO Feng, XU Xing lian, et al. Changes in the bacterial communities of vacuum-packaged pork during chilled storage analyzed by PCR-DGGE[J]. Meat Science, 2010, 86: 889-895.
[12] JIANG Yun, GAO Feng, XU Xinglian, et al. Changes in the composition of the bacterial flora on tray-packaged pork during chilled storage analyzed by PCR-DGGE and real-time PCR[J]. Journal of Food Science, 2011, 76: M27-M33.
[14] 李苗云. 冷却猪肉中微生物生态分析及货架期预测模型的研究[D]. 南京农业大学博士论文,2006.
[13] LI Miaoyun, ZHOU Guanghong, XU Xinglian, et al. Changes of bacterial diversity and main flora in chilled pork during storage using PCR-DGGE[J].Food Microbiology，2006,23（7）：607-611.
[15] 田永全.冻结肉、热鲜肉、冷鲜肉的营养与卫生[J].中国食物与营养， 2007,5:61-62.
[16] SMALL A A，WELLS B B,BUNCIC A S,et al. An evaluation of selected methods for the decontamination of cattle hides prior to skinning[J].Meat Science,2005,69,297-314.
[17] GILL C O,DUSSAULT F,HOLLEY R A,et al. Evaluation of the hygienic performances of the processes for cleaning,dressing and cooling big carcasses at eight picking plants[J].International Journal of Microbiology,2000,58:65-72.
[18] DAINTY J M,SHERIDANA J J,BLAIRB I S,et al. Microbial contamination on beef in relation to hygiene and assessment based on criteria used in EU Decision 2001/471/EC[J]. International Journal of Microbiology,2004,92:217-225.
[19] DAINTY R H,MACKEY B M. The relationship between the phenotypic properties of bacteria from chill-stored meat and spoilage processes[J].Journal of Applied Bacteriology Symposium Supplement,1992,73:103-114.
[20] OLSSON C,AHME B,MOLINA G.The bacterial floral of fresh and chill-stored pork: analysis by cloning and sequencing of 16SrRNA genes[J]. International Journal of Microbiology,2003,83:245-252