Spoiled meat niche

Description of Niche

Where located?

Spoiled meat varies in different forms, from being raw, marinated, or being cooked and cured. The spoiled meat is stored in a vacuum or modified atmosphere.[1SL] Spoiled meat can usually be found in refrigerators at low temperatures.

Physical Conditions?

What are the conditions in your niche? Temperature, pressure, pH, moisture, etc.

Because of its significant nutrients available on the surface and its high water content, meat is one of the most perishable foods. The conditions of spoiled meat vary on the type of meat, how the meat was prepared, and how the meat is stored. Pork The follow conditions are for Frankfurter-type sausages and sliced pork. For meat that is initially cooked/heated at 65-75º C [2SL] and stored in a vacuum or modified atmosphere, at a temperature of 4º C, the salt content is around 2%, with a pH of above 6, making it a generally neutral environment. The moisture is 61.5% in Frankfurter-type sausages and 68% in sliced pork. [1SL] Chicken Marinated chicken meat in a modified atmosphere (MA) packaged and stored at temperatures ranging from 3.4 to 7.7ºC. The marinating factor includes many sub-ingredients such as acidic sauce including sucrose or glucose, salt, and spices that adds to the flavor of meat. [1SP] The marinate provides an acidic pH of 4.7 to 5.0. [1SP] Beef For ground beef (fresh from the market) stored aerobically (high oxygen MA) at the temperature of 5-7ºC, chilled- refrigerated, the pH was 5.5 to 5.92. [2SP] For raw beef steaks, the pH was 5.4 to 5.6 the lactic acid bacteria level was 2.2 × 104 CFU/g [4SP]. The beef steak meat packages were also moisture enhanced with levels between 112 and 115% of original weight [4SP].

Influence by Adjacent Communities (if any)

Is your niche close to another niche or influenced by another community of organisms?

Because it is stored in a modified, vacuum, refrigerated atmosphere, the spoiled meat niche is not close to another niche, but the handling and process of packaging meat and can often add microflora to the products. An estimated .5-2 log cfu/g of bacteria is added to the meat due to these processes. [2SL]

Conditions under which the environment changes

Do any of the physical conditions change? Are there chemicals, other organisms, nutrients, etc. that might change the community of your niche.

The influenced on spoiled meat is by the microflora on the meat itself, mainly Lactobacillus sp. and Leuconostoc sp., and human added Lactic acid bacteria. Lactic acid bacteria are antagonistic cultures added to the meat in order to inhibit pathogens and increase the shelf life, which are protective cultures as a means of biopreservation. In this way, Lactic acid changes the condition of the niche by inhibiting unwanted microorganisms, via several methods, such as the production of bacteriocins and enzymes, and also by simply competing for nutrients in the same niche.

The niche community changes as the shelf life continues. The presence of lactic acid bacteria can prevent spoilage flora from growing. [1SL] If stored properly, either refrigeration or an anaerobic atmosphere, lactic acid bacteria will prevent other microflora from spoiling the meat, and lactic acid bacteria will itself dominate the spoilage process. [2SL] As the shelf life reaches its limit, Lactic acid bacteria populations increase which cause spoilage. Depending on the product handling after cooking, the spoiled pork may be recontaminated with up to .5-2 log CFU/g of total bacteria, which are mainly lactic acid bacteria. [2SL]

In frankfurter-type sausages, the presence of Lact. curvatus can potentially change the physical conditions of the niche by decreasing the pH of the spoiled meat (from 6.6 to 5.8) at the end of its shelf life, after 28 days. In pork, the presence of Lactic Acid bacteria caused a decrease of pH from 6.6 to 5.1-5.3 (vacuum) and 5.5-5.6 (modified atmosphere) at the end of its shelf life. [1SL]

The conditions of the niche consists can cause various changes ranging from souring, changing the flavor, texture, and color, producing gas and slime, and change in the pH level. Spoilage affects can be noticed at various points in the shelf life, as early as a few days and up to the end of the shelf life (18 to 42 days), where the meat is dominated by spoilage. [3SL] Meat often shows a distinct color change from a reddish meat color to a brown pigment and shows bulging due to gas formation after packaging. [3SP] Spoilage is also apparent by swelling of packages and characterized by a green coloring due to production of hydrogen peroxide [4SP] with a strong buttery odor due to putrescine and cadaverine. [1SP] In addition, samples of meat loses water during storage (weight loss). Slime formation may occur, such as biofilms at low temperatures and have quorum sensing which also produces pigment changes that may not be suitable for human consumption. The development of spoilage is also associated to microbial consumption of meat nutrients such as sugars and amino acids. [3SP]

The influence of Chemicals
Increasing concentrations of CO2, a decreased pH, and a chilled storage are used to preserve meat and prevent spoilage microflora from forming [2SP]. But still, the dominant organisms Lactobacillus oligofermentans and Pseudomonas are present.

Marinating chicken often maintains the low pH because of the low pH of the marinate. The process of marinating in chicken can support the growth of Lactobacillus oligofermentans in which its population can grow up to 10^8 to 10^10 CFU/g. [1SP] Ingredients also used to promote flavoring and taste of beef/steak and vacuum packaging with glucose and sugar can expedite the spoilage process that is depicted through a green coloring and a strong buttery odor of the meat. [4SP]

In the experiment Spoilage of value-added, high-oxygen modified-atmosphere packaged raw beef steaks by Leuconostoc gasicomitatum and Leuconostoc gelidum, the effects of O2 and CO2 content was studied via three packaging conditions of beef. All conditions were modified atmosphere packaged but with different concentrations of O2 and CO2: MAP1 consists of only air; MAP2 is 60% O2 and 40% CO2; and MAP3 is 20% O2 and 40% CO2. [3SP] These three varying packaging conditions show different effects on the spoiling piece of meat. The conditions for MAP2 protect the spoilage of meat (little color change of the beef and little microbial loads) after refrigerated storage compared to MAP1 and MAP3. According to the findings of different packaging condition studies, many different changes occurred. For MAP1 where only oxygen is present, the concentration of O2 decreased from 21% to 0% and concentrations of CO2 increased from 0% to 25% in 14 days.[3SP] In the MAP2 condition, both concentrations of O2 and CO2 remain constant. [3SP] For MAP3, the same observations is seen for the first seven days, but during the next seven days (day 7-14), a drop in oxygen concentration and an increase in CO2 concentration is detected. [3SP]

Who lives there?

Which microbes are present?

Brochotrix thermosphacta
Brochothrix Thermosphacta is a microorganism for which meat is considered an ecological niche. Its ability to grow under both aerobic and anaerobic conditions makes a significant meat colonizer. The genus Brochothrix is characterizes as gram-positive, nonsporeforming, nonmotile, catalase-postiive, facultatively anaerobic, reluglar, rod shaped bacteria.. The optimal temperature for growth is 20-25º C. The optimal pH for B. thermosphacta to grow is pH 7.0 but growth is seen within the ranges of pH 5-9. Brochothrix thermosphacta is more resistant to irradiation than common meat spoilage organism such as Pseudomonas but are affected by irradiation does of 0.5 to 2.0 kilogray. The species have often been isolated from irradiated meat and poultry. Although, they are an important spoilage organism found prepacked meats and in meat stored in chill temperature, they can also inhabit other niches such frozen foods, milk, and cream. Storage conditions often selectively favor its growth. Brochothrix ssp can grow at temperatures a low as 0º C and under conditions of low oxygen concentration and high C02 concentration. For metabolism, Brochothrix thermosphacta has enzymes for both the hexose-monophosphate and glycolysis pathways of glucose. Fermentative metabolism of glucose always results in the production of L+ lactic acid, but other end products depend on growth conditions [8]. Major end products of aerobic metabolism of glucose by B. Thermosphacta growing on meat are acetoin and acetic, isobutyric, isovaleric and 2 methylbutyric acis. In minimal medium, glucose is the source of all the end products; However, in complex medium such as meat, only acetoin and acetic acid are derived from glucose; Isobutyric, isovaleric, and 2-methylbutyric acids are produced from valine, leucine, and isoleucine, respectively[s4]. These compounds, or their derivatives, are responsible for the odor that often characterizes spoiled meat. Unlike proteolytic spoilage bacteria such as Pseudomonas, B. thermosphacta is usually found only on the meat surface. In prepacked meat, it grows in the area between the meat-plastic film [s8].

Carnobacterium

Clostridium [[1]]
Clostridium is a rod-shaped cell with a gram-positive membrane. These microbes are anaerobes and some are toxin-producing pathogens. Some of them produce acetone, butanol, ethanol, isopropanol, and organic acids. This bacterium can go through spore formation for survival. Clostridium produces large amounts of gas in packaged meat. It is usually coupled up with foul odors and causes the package to appear in a blown pack. [JT5] The toxin produced by this bacterium can do harm and help heal. So far this toxin has helped treat dystonias (neurologic diseases involved abnormal muscle posture and tension), urinary bladder muscle relaxation, esophageal sphincter muscle relaxation, and tics. However at the same time, the toxin released can cause botulism poisoning. Proteolytic strains of toxin is produced at around 35°C and for nonproteolytic strains, they can grow in environments of 26-28°C. Toxin produced from bacterium will cause botulism which is food poisoning that will lead to muscle paralysis. [JT6]

Enterobacterium

Lactobacillus [[2]]

Leuconostoc [[3]]
Leuconostoc is one of the lactic acid bacteria; it produces D-lactate and ethanol. This group of microbe is responsible for the discoloration, gas production, and buttery smell of spoiled meat. [JT2] The genus Leuconostoc is described as being spherical cells that is gram-positive and often lenticular on agar. This bacterium grows optimally in an environment of 20-30°C and in modified atomospheres. However, they also require a rich and complex media for growth. A rich and complex media includes nicotinic acid, thiamin, biotin, and pantothenic acid. For energy, they are heterofermentatives, which means they use a combination of pentose phosphate and phosphoketolase pathways. This microbe cannot go through spore formation for survive. They fall under the facultative anaerobic category, which means they can live in an environment with or without oxygen. Leuconostoc was originally placed into Streptococcaceae bacteria family as mentioned in Bergey’s Manual of Determinative Bacteriology. [JT4] However, in 1986, the Bergey’s Manual of Systematic Bacteriology moved Leuconostoc from the Streptococcaceae family into the Deinococcaceae family. [JT4] The green spots on a slice of spoiled meat are caused by the H2O2 created by Leuconostoc. Leuconostoc mesenteroides, Leuconostoc carnosum, and Leuconostoc amelibiosum are responsible for the accumulation of CO2 production. [JT4] Aside from finding Leuconostoc in spoiled meat, it can also grow in plants, fermenting vegetables, milk, dairy products, wine, and even human blood. [JT4]

Pseudomonas [[4]]

The predominant bacteria that are often associated with spoiled meat are Pseudomonas. They are polarly flagellated, gram –negative, rod shaped, aerobic bacteria. [a1]. A few microorganisms under the genus Pseudomonas are known to effectively use meat as a niche due to their ability to break down glucose and amino under aerobic conditions and at refrigerated temperature. Pseudomonades are able to break down the long peptide chains of proteins in meats into amino acids and foul-smelling compounds such as ammonia, amines, and hydrogen sulfide [a2]. Some strain of Pseudomonas produce esters, many produce sulfur-containing compounds, and a few produce methyl ketones, secondary alcohols, and unsaturated hydrocarbons [a3]. Florescent Pseudomonas strains represent one of the most important groups among Pseudomonas because of their ability to produce water-soluble yellow-green pigments, called pyoverdines (PVDs). These yellow-green pigments act as siderophores, allow Pseudomonas to uptake iron from their environment. The most common Pseudomonas species found in beef, pork, lamb and poultry meat appears to be Pseudomonas fragi. Perhaps Pseudomonas fragi strains are so dynamic because it is capable of using a wide range of carbon compounds including D-arabinose, creatine, and bile acids [a6]. Pseudomonas fragi growing on meat surface uses compounds such as glucose, free amino acids, and lactate. These carbon sources are enough support growth until spoilage has occurred. When the concentration of these compounds decrease in the uppermost layer, the compounds diffuse from below. Proteolytic activity and penetration of bacteria down in the tissue does not occur before the meat is already spoiled. In general, Pseudomonas shows preference for glucose. It is only when glucose is depleted that the Pseudomonas takes up the free amino acids (the amino acids are consumed before lactate). The order of preference from most to least is glucose> lactate>citrate>aspirate-glutamate>creatine-creatinine. It is at the point when amino acids are consumed that the meat gives off an offensive odor from the volatile by-products of amino acid catabolism. [a7].

Shewanella putrefaciens

Do the microbes change their environment?

Brochothrix thermosphacta, Carnobacterium spp., Enterobacteriaceae, Lactobacillus spp., Leuconostoc spp., Pseudomonas spp., Shewanella putrefaciens and Weissella spp. work together to create the spoiled meat profile: discoloration, gas production, slime production, decrease in pH, and sour off-flavor. [JT2]

Leuconostoc produces H2O2, which gives spoiled meat its green discoloration. [JT7]

Clostridium work with lactic acid bacteria [Lactobacillus and Leuconostoc] to produce large amounts of gas (H2 and CO2) which is accompanied by a foul odor. [JT2]

Do the microbes carry out any metabolism that affects their environment?

Clostridium can perform nitrogen fixation. Clostridium can go through fermentation of carbon sources to produce acetone, butanol, ethanol, isopropanol, and organic acids. [JT5]

Leuconostoc produces ammonia by the use to bacterial deamination of amino acid and the production of ammonia will lead to a decrease in acidity. The process it takes to produce H2O2 involves the oxidation of nitrosohaemochrome to choleomyoglobin. [JT7]

Current Research

2006: lactic acid bacteria associated with vacuum-packed cooked meat product spoilage: population analysis by rDNA-based methods
The investigators aimed to research and find which lactic acid bacteria was involved in the spoilaged of vacuum packaged cooked meat products. They did this by studying different samples of bacteria within 4 meat products, some of which had spoilage symptons, some that did not. Colonies of these were then grown on yeast glucose lactose peptone and trypticase soy yeast plates, and where then identifived via internal spacer region. The study found that Leuc. Mesenteroides was the main spoilage agent within vacuum packaged meats. The significance of this study was to determine what organisms to look for to prevent the spoilage of vacuum packaged meats. [1]

2005: Development of a Microbial Model for the Combined Effect of Temperature and pH on Spoilage of Ground Meat, and Validation of the Model under Dynamic Temperature Conditions
The study aimed at using microbiological and sensory analysis to predict spoilage of aerobic stored ground meat. Under aerobic conditions, samples of ground meat (beef and pork) were analyzed for changes in their appearances, smells and microbes composition at certain ranges of pH (5.34-6.13) and temperature (0-20 Celcius). As observed, pseudomonads were the predominant bacteria isolated from these samples. In addition, it was also detected that the changes in pseudonomads populations is proportional to the sensory changes. Thus,it can be concluded that microbiological and sensory analysis can be used as a “good index for spoilage of aerobically stored ground meat”. Following this type of model, the meat industry can benefit from by running more “effective management systems, which will optimize the quality of meat products”. [3 JN]

2003: In vitro and in situ growth characteristics and behaviour of spoilage organisms associated with anaerobically stored cooked meat products
This study was aimed to research the different types of spoilage affects caused by different organsisms, in vacuumed packaged cooked meat products. They did this by characterizing strains of different spoilage organisms in a 7°C anaerobic broth. The growth rate, acidifying character, and metabolite production was compared. Then the organisms were inoculated onto cooked meat, and characterstics were again obsereved, including spoilage, microbial growth, pH, metabolite production, and also sensory changes. The results concluded that the microbial organisms B. thermosphacta and Leuc. Mesenteroids induced spoilage on cooked meat products the quickest. This study determined correlations between microbial growth, changes in pH and metabolite formation and various spoilage organisms upon cooked ham. [10]

2002: Microbial interaction in cooked cured meat products under vacuum or modified atmosphere at 4º C
This study was aimed to find out the antagonistic activity of the lactic acid bacteria strains Leuconostoc mesenteroides and Lactobacillus curvatus against spoilage in two types of cooked meat. The results concluded that the lactic acid did help prevent spoilage while they did not negatively affect the meat. The study found that biopreservation can help increase the shelf life of meat produce. [11]

References

[1] Chenoll, E., Macian, M., Elizaquivel, P., Aznar, R. "Lactic Acid Bacteria Associated with Vacuum-packed Cooked Meat Product Spoilage: Population Analysis by rDNA-based Methods". Journal of Applied Microbiology. 2006. Volume 102. p. 498-508.

[2] Borch, E., Kant-Muermans, M., Blixt, Y. "Bacterial Spoilage of Meat and Cured Meat Products". International Journal of Food Microbiology". 1996. Volume 33. p. 103-120.

[3] Koutsoumanis, K., A. Stamatiou, P. Skandamis, and G.-J. E. Nychas. "Development of a Microbial Model for the Combined Effect of Temperature and pH on Spoilage of Ground Meat, and Validation of the Model under Dynamic Temperature Conditions". Applied and Environmental Microbiology. 2005. Volume 72. p. 124-134.

[4] Thunel, R. "Taxonomy of the Leuconostocs" Journal of Dairy Science. 1995. Volume 78. no. 11. P. 2514-2522

[5] Nolling J., Breton G., Omelchenko M.V., Makarova K. S., Zeng Q., Gibson R., Lee H. M., Dubois J., Qui D., Hitti J., Wolf Y. I., Tatusov R. L., Sabathe F., Doucette-Stamm L., Soucaille P., Daly M.J., Bennett G.N., Koonin E. V., Smith D. R. "Genome sequence and comparative analysis of the solvent-producing bacterium Clostridium acetobutylicum". Journal of Bacteriology. 2001 Volume 183. no. 16. p. 4823-38.

[6] Vangelova, L. “Botulinum Toxin: A Poison That Can Heal”. FDA Consumer Magazine. 1995.

[7] [Koort, J., Murros, A., Coenye, T., Eerola, S., Vandamme, P., Sukura, A., Bjorkroth, J. “Lactobacillus oligofermentans sp. Nov., Associated with Spoilage Modified-Atmosphere-Packaged Poultry Products”. Applied and Environmental Microbiology. 2005. Volume 71. No 8. p. 4400-4406]

[8] Jorngen J. Leisner, B. G. Laursen, H. Prevost, D. Drider, and P. Dalgaard. 2007. Carnobacterium: Positive and Negative Effects in the Environment and in Foods. FEMS Microbiol Rev. 31: 592-613.

[9] Weigand I., H. K. Geiss, D. Mack, E. Sturenburg, and H. Seifert. 2007. Detection of Extended-Spectrum Beta-Lactamases among Enterobacteriaceae by Use of Semiautomated Microbiology Systems and Manual Detection Procedures. Journal of Clinical Microbiol. 45: 1167-1174.

[10] [Vermeiren, L., Devlieghere, F., De Graef, V., Debevere, J. "In vitro and in situ growth characteristics and behaviour of spoilage organisms associated with anaerobically stored cooked meat products". Journal of Applied Microbiology. 2003. Volume 98. p. 33-42.]

[11] [Metaxopoulos, J., Mataragas, M., Drosino, E. "Microbial interaction in cooked cured meat products under vacuum or modified atmosphere at 4º C". Journal of Applied Microbiology. 2002. Volume 93. p. 363-373.]

[12] [[5] Anzai Y, Kim H, Park, JY, Wakabayashi H (2000). "Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence". Int J Syst Evol Microbiol 50: 1563–89.]

Edited by [Steven Lee , Jade Nguyen , Ngoc-minh Nguyen , Sarah Paek , June Tse , Amy Vo], students of Rachel Larsen