Effects of temperature, oxygen exclusion, and storage on the microbial loads and pH of packed ostrich steaks

Ostrich steaks (96) were packed (air or vacuum) and stored at 4°C or 10°C. Microbiological loads (total viable counts - TVC, psychrotrophic, Enterobacteriaceae, lactic acid bacteria - LAB, Pseudomonas and fluorescent Pseudomonas) and pH values were determined at 0, 3, 6 and 9 days. High counts (8.0-10.2 log(10)cfu TVC/g) were observed at day 9, possibly as a consequence of the initial high microbial load (range 4.9-5.4 log(10)cfu TVC/g) and pH (average 6.7). Temperature and sampling day significantly influenced all microbial counts. Gas atmosphere had a small or negligible influence on levels of Enterobacteriaceae and LAB. For most microbial groups temperature significantly influenced bacterial levels up to day 6 of storage, while gas atmosphere had a significant effect at days 6 and 9. Both effects (temperature and gas atmosphere) were significant factors from day 0 for fluorescent Pseudomonas. Samples vacuum-packed and stored at 4°C showed the lowest microbial loads at day 9. Only for these samples was no sensory rejection observed at the end of the experiment.     

Control of Listeria monocytogenes on ham steaks by antimicrobials incorporated into chitosan-coated plastic films

Contamination of ready-to-eat (RTE) meat products such as ham steaks with Listeria monocytogenes has been a concern for the meat processing industry. The objective of this study was to evaluate the antilisterial efficacy of chitosan-coated plastic films alone or incorporating five generally recognized as safe (GRAS) antimicrobials. Effect of chitosan-coated plastic film on the growth of L. monocytogenes was first investigated in an aqueous system of culture medium broth and chitosan-coated films were able to inhibit the growth of L. monocytogenes in a concentration-dependent manner. However, chitosan-coated plastic films were not able to control the growth of L. monocytogenes on ham steaks. Therefore, five GRAS antimicrobials were subsequently incorporated into chitosan-coated plastic films to enhance their antilisterial effectiveness. Ham steaks were surface-inoculated with a five-strain cocktail of L. monocytogenes and then packaged in chitosan-coated plastic films containing 500 IU/cm(2) of nisin, 0.01 g/cm(2) of sodium lactate (SL), 0.0025 g/cm(2) of sodium diacetate, 0.003 g/cm(2) of potassium sorbate (PB), or 0.001 g/cm(2) of sodium benzoate (SB). The samples were stored at room temperature (ca. 20 degrees C) for 10 days. Incorporating antimicrobials into chitosan-coated plastic films slowed down or inhibited the growth of L. monocytogenes. The chitosan-coated plastic film containing SL was the most effective antimicrobial film and its efficacy against L. monocytogenes on ham steaks was evaluated during 12-week storage at 4 degrees C. The film showed excellent long-term antilisterial effect with the counts of L. monocytogenes being slightly lower than the initial inoculum. Chitosan-coated plastic films containing 0.001 g/cm(2) of SL have a potential to be used on ham steaks to control L. monocytogenes.     

Effect of a bacteriocin produced by Pediococcus acidilactici against Listeria monocytogenes and Clostridium perfringens on Spanish raw meat

The inhibitory effect of a bacteriocin, produced by Pediococcus acidilactici, against Listeria monocytogenes and Clostridium perfringens on Spanish raw meat surface, was evaluated by in situ assays. Samples were incubated with the bacteriocin and then with a culture of the pathogenic bacteria. The treatment with 500, 1000 or 5000 bacteriocin units/ml (BU/ml) reduced the counts of L. monocytogenes after storage at 15°C during 72h by 1, 2 or 3 log cycles and with 1000 or 5000 BU/ml after storage at 4°C during 21 days by 2.5 or 3.5 log cycles, respectively, compared to the control. With C. perfringens a bacteriostatic effect could be observed.     

Use of antimicrobial biodegradable packaging to control Listeria monocytogenes during storage of cooked ham

The antimicrobial effect against L. monocytogenes of biodegradable films (alginate, zein and polyvinyl alcohol) containing enterocins was investigated. Survival of the pathogen was studied by means of challenge tests performed at 6 degrees C during 8 and 29 days, for air-packed and vacuum-packed sliced cooked ham, respectively. Air packaging was tested with two concentrations of enterocins (200 and 2000 AU/cm2). Control air-packed cooked ham showed an increase of L. monocytogenes from 10(4) to 10(7) CFU/g after 8 days. By contrast, packaging with antimicrobial films effectively slowed down the pathogen's growth, leading to final counts lower than in control lots. Air-packaging with alginate films containing 2000 AU/cm2 of enterocins effectively controlled L. monocytogenes for 8 days. An increase of only 1 log unit was observed in zein and polyvinyl alcohol lots at the same enterocin concentration. Vacuum packaging with films containing enterocins (2000 AU/cm2) also delayed the growth of the pathogen. No increase from inoculated levels was observed during 15 days in antimicrobial alginate films. After 29 days of storage, the lowest counts were obtained in samples packed with zein and alginate films containing enterocins, as well as with zein control films. The most effective treatment for controlling L. monocytogenes during 6 degrees C storage was vacuum-packaging of sliced cooked ham with alginate films containing 2000 AU/cm2 of enterocins. From the results obtained it can concluded that antimicrobial packaging can improve the safety of sliced cooked ham by delaying and reducing the growth of L. monocytogenes.     

Edible chitosan films on ready-to-eat roast beef for the control of Listeria monocytogenes

The use of chitosan as an edible film was evaluated for its antimicrobial activity against Listeria monocytogenes (LM) on the surface of ready-to-eat (RTE) roast beef. L. monocytogenes, decimally diluted to give an initial inoculation of >6.50logCFU/g, was inoculated onto the surface of RTE roast beef cubes, and air-dried. The samples were dipped into chitosan (high or low molecular weights) solutions dissolved with acetic or lactic acid at 0.5% (w/v) or 1% (w/v) then bagged and refrigerated at 4 degrees C. The bacterial counts were determined on days 0, 7, 14, 21, and 28. The samples were spread plated onto modified Oxford agar plates and incubated at 37 degrees C for 48h. An initial 6.50logCFU/g of L. monocytogenes inoculated onto the surface of the non-coated RTE roast beef increased too >10logCFU/g by day 28. On day 14, L. monocytogenes counts were significantly different for all the chitosan-coated samples from the control counts by 2-3logCFU/g and remained significantly different on day 28. Our results have shown that the acetic acid chitosan coating were more effective in reducing L. monocytogenes counts than the lactic acid chitosan coating. Our study indicated that chitosan coatings could be used to control L. monocytogenes on the surface of RTE roast beef.     

Reduction of counts of Listeria monocytogenes in cheese by means of high hydrostatic pressure

Inactivation of Listeria monocytogenes (strains NCTC 11994 and Scott A) was evaluated in model cheeses submitted to 10 min HHP treatments of 300, 400 or 500 MPa at 5 or 20 degrees C. Counts were measured immediately after high hydrostatic pressure (HHP) treatment (day 1) and after 2, 15 and 30 days of storage at 8 degrees C. Both strains behaved significantly different after 400 and 500 MPa, being NCTC 11994 more sensitive. Scarce differences were found among final values at both HHP treatment temperatures. Initial reductions (log cfu/g) for 400 MPa at 20 degrees C were 2.9 +/- 0.2 for strain NCTC 11994 and 1.5 +/- 0.2 for Scott A. They reached after 30-day storage 5.3 +/- 0.2 and 4.6 +/- 0.4 log cfu/g for NCTC 11994 and Scott A, respectively. For 500 MPa treatments, day-1 reductions of both strains were around 5-log cfu/g, and counts fell below quantification limit after 30 days. Injured cells (around 0.8-log cfu/g) were mostly observed in 400 MPa treated samples on days 1 and 2. Starter cells suffered higher inactivation and injury. For 20 degrees C treatments, its final counts (log cfu/g) at 300, 400 and 500 MPa were: 8.5 +/- 0.2, 5.4 +/- 0.3 and 2.5 +/- 0.1, respectively. These figures evidence the HHP potential to improve safety of cheese products.     

Evaluation of the storage life of vacuum packaged Australian beef

To establish the shelf-life of vacuum packaged Australian beef, 15 Strip-loins and 15 Cube-rolls for each Processor (A, B, and C) were evaluated at two week intervals (since wk 10 to 20). Steaks on the trays were placed in retail cabinets at 3 °C. Shelf-life evaluation was based on off-odor (only at week 10), microbial analysis, lipid oxidation, and color assessment by trained panelist and Hunter colorimeter. Panelists detected "slightly off-odor" in both primal cuts for Processors B and C (P<0.05). Processor A primal cut steaks displayed better color scores as well as CIE L*, a*, b*, Chroma and Hue values during storage and display period than steaks from the other processors. Also, primal cuts from Processor A showed lower microbial counts and TBARS values with respect to other processors during the trial. Processor A cuts showed improved shelf life attributes initially which helps to explain its slow shelf-life deterioration.     

Shelf life of ostrich (Struthio camelus) liver stored under different packaging conditions

Ostrich (Struthio camelus) liver is an important edible meat by-product. However, it has a low commercial value, and it is underutilized. The present study was conducted to establish the shelf life of ostrich liver, stored at 2 +/- 1 degrees C for 18 days, under different packing conditions. Fresh livers were sliced and packed under air exposure, vacuum, and modified atmosphere (MA: 80% CO2 plus 20% N2). Shelf-life evaluation was based on color, lipid and hemopigment oxidation, microbial counts, and sensory assessment of odor and color. Samples stored under air exposure showed the highest lipid and hemopigment oxidation rate, which was detected by higher thiobarbituric acid reactive substances values and metmyoglobin percentage and lower heme iron content than livers stored under vacuum or MA. On the basis of aerobic bacteria counts, the shelf life of ostrich liver stored under aerobic conditions would be 7 days, whereas under vacuum or MA, it would be 14 days. In livers stored under anaerobic conditions (vacuum or MA packed), lactic acid bacteria represented a significant proportion of the spoilage microbiota. Minimal changes in color perception were also obtained in vacuum-packed samples at the end of the storage period. In air-packed samples, high levels of off-odors were already perceived during the first 6 days of storage, whereas in livers stored under anaerobic conditions (vacuum and MA), these levels of off-odor were not yet perceived at that time.     

The fate of two Listeria monocytogenes serotypes in "cig kofte" at different storage temperatures

Cig kofte is a traditional Turkish food prepared from minced beef, bulgur, onions, garlic and varieties of spices. It is generally consumed within a few hours. However, leftovers can be kept in refrigerator or in room temperature up to 24h until they are consumed. In this study, survival and growth of two Listeria monocytogenes serotypes were investigated in cig kofte during the storage. For this purpose, the prepared samples were separately contaminated with serotypes 1/2b or 4b of L. monocytogenes at the level of 10(4)CFU/g and stored at 4°C and 21°C. L. monocytogenes colonies were counted at the beginning, 3rd, 6th, 12th and 24th hours of the storage. At 4°C, L. monocytogenes 4b significantly increased (P<0.05) from 4.12 to 5.49log(10)CFU/g but L. monocytogenes 1/2b remained constant (P>0.05) during the storage period. At 21°C, both L. monocytogenes 1/2b and 4b increased significantly (P<0.05) from 4.56 to 5.16log(10)CFU/g and from 4.23 to 5.65log(10)CFU/g, respectively. The physicochemical and microbiological characteristics of the cig kofte did not inhibit the growths of L. monocytogenes serotypes during the storage. These results indicated that L. monocytogenes was able to survive and grow in cig kofte at the both storage temperatures of 4°C and 21°C and cig kofte seemed to be a suitable medium for this pathogen.     

Refrigerated shelf life of vacuum-packaged, previously frozen ostrich meat

Previously frozen ostrich meat was evaluated over 28 days to determine the refrigerated shelf life. Intact steaks and ground meat from three ostrich carcasses were vacuum-packaged, frozen to -40°C for 5 days, and stored in a 0°C walk-in cooler. Instrumental analysis of CIE L*a*b* values indicated that ostrich meat was very dark in color, initially and over time. Microbial growth stayed slightly below 1.0 × 10(7) CFU/g for up to 21 days of refrigerated storage. Sensorially evaluated color showed an increase (p <0.05) in darkness over time. Percentage of browning increased (p<0.05) over time from 1% initially to 55% for intact steaks and 75% for ground meat by 28 days. Sensory aroma scores significantly (p<0.05) changed over time, with unacceptable aroma occurring by 14 days. Previously frozen, vacuum-packaged ostrich meat stored under refrigerated conditions should be used within 10 days.     

Effect of high pressure treatment on microbial populations of sliced vacuum-packed cooked ham

In this study, culture-dependent and culture-independent approaches were used to reveal the microbial diversity and dynamic changes occurring in sliced vacuum-packed cooked ham after high pressure processing (HPP, 400MPa or 600MPa for 10min at 22°C) during refrigerated storage over 90days. Direct extraction of genome DNA and total RNA from meat samples, followed by PCR-denaturing gradient gel electrophoresis (DGGE) and RT-PCR-DGGE on 16S rDNA V3 region, was performed to define the structure of the bacterial populations and active species in pressurized cooked ham. Results showed that HPP affected differently the various species detected. The predominant spoilage organisms of cooked ham, such as Lactobacillus sakei and Lactobacillus curvatus, were found to be very sensitive to pressure as they were unable to be detected in HPP samples at any time during refrigerated storage. Weissella viridescens and Leuconostoc mesenteroides survived HPP at 600MPa for 10min at 22°C and were responsible for the final spoilage. An RNA-based DGGE approach clearly has potential for the analysis of active species that have survived in pressurized cooked ham. High pressure processing at 400 or 600MPa for 10min at room temperature (22°C) has a powerful inhibitory effect on the major spoilage bacteria of sliced vacuum-packed cooked ham. High pressure treatment may lead to reduced microbial diversity and improve the products' safety.     

Effects on the development of blown pack spoilage of the initial numbers of Clostridium estertheticum spores and Leuconostoc mesenteroides on vacuum packed beef

Beef steaks were inoculated with Clostridium estertheticum spores and Leuconostoc mesenteroides cells at all combinations of numbers of 0, 10, 100 or 1000/cm(2) for each organism. After vacuum packaging the steaks were stored at 4, 1, or -1.5°C. Pack volumes were determined by water displacement at suitable intervals. Irrespective of L. mesenteroides numbers, for packs containing meat inoculated with 0, 10, 100 or 1000 spores/cm(2), 60, 16, 3 and 1 of 60 packs did not swell. The times of onset of swelling were twice as long at -1.5 as at 4°C, but they were not affected by the inoculated numbers of L. mesenteroides. Rates of pack swelling increased with increasing storage temperature and number of spores, but decreased with increasing numbers of inoculated L. mesenteroides. Lactic acid bacteria can apparently prevent development of blown pack spoilage of vacuum packs containing meat contaminated with low numbers of C. estertheticum.     

High-pressure processing and antimicrobial biodegradable packaging to control Listeria monocytogenes during storage of cooked ham

The efficiency of combining high-pressure processing (HPP) and active packaging technologies to control Listeria monocytogenes growth during the shelf life of artificially inoculated cooked ham was assessed. Three lots of cooked ham were prepared: control, packaging with alginate films, and packaging with antimicrobial alginate films containing enterocins. After packaging, half of the samples were pressurized. Sliced cooked ham stored at 6 degrees C experienced a quick growth of L. monocytogenes. Both antimicrobial packaging and pressurization delayed the growth of the pathogen. However, at 6 degrees C the combination of antimicrobial packaging and HPP was necessary to achieve a reduction of inoculated levels without recovery during 60 days of storage. Further storage at 6 degrees C of pressurized antimicrobial packed cooked ham resulted in L. monocytogenes levels below the detection limit (day 90). On the other hand, storage at 1 degrees C controlled the growth of the pathogen until day 39 in non-pressurized ham, while antimicrobial packaging and storage at 1 degrees C exerted a bacteriostatic effect for 60 days. All HPP lots stored at 1 degrees C led to counts <100CFU/g at day 60. Similar results were observed when combining both technologies. After a cold chain break no growth of L. monocytogenes was observed in pressurized ham packed with antimicrobial films, showing the efficiency of combining both technologies.     

Microbiological shelf life of bulk-packaged Musculus gluteus medius supplemented with dietary vitamin E

Five Simmentaler type calves were fed diets supplemented with 500 mg vitamin E per day and five fed control diets. Rump steaks from each carcass were PVC-overwrapped and bulk packaged in 100% CO, or 20% CO(2):80% O(2). Bulk packs were stored up to 42 days at 4°C and steaks displayed up to 7 days at 4°C. Bacterial counts of rump steaks from either packaging treatment were not significantly influenced during bulk storage or retail display by supplementation with dietary vitamin E. Both packaging treatments delayed bacterial growth during bulk storage. Aerobic plate counts of rump steaks stored in 100% CO(2) were lower than those of rump steaks stored in 20% CO(2): 80%: O(2). This study showed that rump steaks supplemented with dietary vitamin E can be bulk packaged in 20% CO(2): 80% O(2) or 100% CO(2) and stored for up to 42 days with shelf life of 4-7 days.     

Enumeration and growth of naturally occurring Listeria spp. in unpackaged ham

A total of 301 unpackaged retail ham samples were tested for the presence and number of Listeria spp. after 7 days at 5 degrees C to simulate domestic storage. Thirteen samples (4.3%) contained Listeria monocytogenes, with the highest count being 1.6 x 10(3)cfu g(-1). Thirteen samples contained other Listeria spp. Genotyping showed that only one L. monocytogenes isolate from the 14 tested was of a type previously identified in New Zealand human cases. Listeria-contaminated batches were incubated at 5 degrees C over approximately 3 weeks to assess the growth rate of natural contaminants. None contained L. monocytogenes, but growth occurred in one sample containing Listeria welshimeri and four containing Listeria innocua. Growth was usually slow at 0.002-0.004 log h(-1). In one sample, L. innocua grew at 0.02 log h(-1) although the maximum number reached was only 4.0-5.0 x 10(3)cfu g(-1). In five other samples little growth, if any, occurred. Growth of naturally occurring Listeria spp. at 5 degrees C was therefore generally slower than predicted by the Pathogen Modelling Programme (PMP) or did not occur.     

Effect of temperature on the spoilage rate of whole, unprocessed crabs: Carcinus maenas, Necora puber and Cancer pagurus

Shelf life of whole, initially live, crabs depended primarily on the storage conditions and the time at which death occurred. Large differences in the time that individual crab species survived particular storage conditions resulted in wide variations in shelf-life. Bacterial spoilage of Carcinus maenas, Necora puber and Cancer pagurus, measured using aerobic plate counts, indicated that on ice at 4 degrees C whole unprocessed crabs had a shelf life approximately 9-11 days, at 4 degrees C approximately 13-29 days, in simulated supermarket conditions of sale approximately 5-7 days and at 20 degrees C approximately 2-16 days. Storage of whole unprocessed crabs chilled at 4 degrees C considerably extended shelf life compared to crabs stored on ice. Live crabs stored on ice died within 24h, most likely due to thermal shock and their early death was responsible for their more rapid increase in bacterial numbers compared to crabs stored at 4 degrees C. No growth of bacteria occurred in the flesh of live crabs stored at 4 degrees C for between 128 and 504 h. Crab flesh quality deteriorated prior to maximum shelf-life (defined as the time at which bacterial load reached log 5 cfu/g crabmeat) in some instances. The best compromise between high crabmeat yield and long shelf-life is likely to be to transport crabs at 4 degrees C live to market where they could be stored live at 4 degrees C without spoilage for 2 weeks before placed on ice at 4 degrees C, with a potential maximum shelf life of approximately 24 days.     

Continued inhibitory effect of carbon dioxide packaging on Listeria monocytogenes and other microorganisms on normal pH beef during abusive retail display

Beef steaks of normal pH (5.3–5.5) were inoculated with Listeria monocytogenes , individually packaged in saturated carbon dioxide atmosphere packaging (SCAP) for <3 h or 5 or 8 weeks or in vacuum packaging (VP) for <3 h, and stored at −1.5°C. After each storage period, 27 individually packaged steaks were removed from their storage packs, overwrapped and placed on retail display under conditions simulating gross temperature abuse (12.25°C). Other steaks were removed from their storage packs and rinsed to remove L. monocytogenes cells, which were re-inoculated onto freshly cut beef steaks to simulate cross-contamination. These crosscontaminated steaks were overwrapped and also subjected to abusive display. Meat pH did not change significantly during storage or retail display. During the retail display of steaks previously stored in SCAP, the lag phase of aerobic bacteria and lactic acid bacteria was longer after prolonged storage compared to short (<3 h) exposure to carbon dioxide. For the same samples, L. monocytogenes failed to grow during retail display or grew only slightly after a prolonged lag phase (>75 h), even after only brief exposure time (<3 h) to carbon dioxide. In contrast, with cross-contaminated steaks, when the inocula had been exposed to SCAP or VP for a short time (<3 h) the L. monocytogenes lag phase was shorter (<20 h). Inocula from steaks stored in SCAP for 5 or 8 weeks did not grow on the cross-contaminated steaks. It is concluded that exposure of both the beef substrate and the L. monocytogenes inoculum to carbon dioxide during prolonged chilled storage does not increase the risk of growth of L. monocytogenes when that meat is subsequently placed on retail display, nor is there a large risk of growth of L. monocytogenes where cross-contamination from SCAP stored raw beef to fresh raw beef occurs prior to retail display.     

Lack of growth of Listeria monocytogenes and Staphylococcus aureus in temperature abuse of E-beam treated ready-to-eat (RTE) cooked ham

The behaviour of Listeria monocytogenes and Staphylococcus aureus in vacuum-packed cooked ham slices treated with an electron beam and stored at 4, 7 and 10 °C was investigated. Cooked ham slices were inoculated with L. monocytogenes and S. aureus and electron beam treated at 2 and 3 kGy. After treatment, a long temperature-dependent death phase was observed, followed by growth at a slower rate than in untreated samples. Assuming a hypothetical load of 10 cells/g or cm² of L. monocytogenes and S. aureus as an original contamination in an industrial situation, an E-beam treatment of vacuum-packed cooked ham slices at 2 kGy guarantees the microbiological safety of the product along its shelf life, even if a noticeable temperature (10 °C) abuse occur during its storage period. Likewise, the E-beam treatment gave rise to a substantial increase of the RTE cooked ham shelf life off-sensory features associated to the spoilage only were detected in non-treated samples (controls) after 8 and 18 days of storage at 10 °C and 7 °C, respectively.     

Acidified sodium chlorite treatment for inhibition of Listeria monocytogenes growth on the surface of cooked roast beef

The effects of acidified sodium chlorite (ASC) against Listeria monocytogenes on the surface of cooked roast beef were investigated. L. monocytogenes, strain V7, serotype 1/2a, was inoculated at numbers of 6.0 log CFU/g onto 5-g cubes of cooked regular or spicy roast beef. The samples were allowed to air dry for 1 h. The cooked roast beef samples were dipped into ASC or sprayed with ASC solutions of 250, 500, 750, or 1,000 ppm, then placed in bags with or without a vacuum and refrigerated at 4 degrees C. L. monocytogenes counts were determined after 0, 7, 14, 21, and 28 days of storage by spread plating roast beef samples onto Oxford agar plates that were incubated at 37 degrees C for 48 h. At day 28, the number of L. monocytogenes on the > or = 500 ppm ASC-treated spicy roast beef samples had count reductions that were >4.0 log CFU/g, whereas the same concentrations of ASC-treated regular roast beef samples had approximately a 2.5 log CFU/g reduction in L. monocytogenes counts when compared with the untreated samples. No significant differences (P > 0.05) were observed in L. monocytogenes counts between the vacuum- or nonvacuum-packaged ASC-treated cooked roast beef samples. Sensory evaluation showed no significant differences (P > 0.05) between ASC-treated and untreated roast beef. ASC can be used as a processing aid in the form of a dip or spray treatment to control L. monocytogenes on the surface of cooked roast beef.