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Changes in distribution patterns and demand for increased food quality have resulted in a desire to improve the shelf life of nonsterile dairy products. Refrigerated shelf life extension typically requires, at a minimum, reductions in the growth rate of spoilage microorganisms and subsequent product deterioration. Reducing initial bacterial loads, increasing pasteurization regimes, and reducing postprocessing contamination have all been employed with measured success. The use of antimicrobial additives has been discouraged primarily due to labeling requirements and perceived toxicity risks. Carbon dioxide (CO2) is a naturally occurring milk component and inhibitory toward select dairy spoilage microorganisms; however, the precise mechanism is not fully understood. CO2 addition through modified atmosphere packaging or direct injection as a cost-effective shelf life extension strategy is used commercially worldwide for some dairy products and is being considered for others as well. New CO2 technologies are being developed for improvements in the shelf life, quality, and yield of a diversity of dairy products, including raw and pasteurized milk, cheeses, cottage cheese, yogurt, and fermented dairy beverages. Here we present a comprehensive review of past and present research related to quality improvement of such dairy products using CO2.

The effect of pressurized (50 MPa) CO2 as a nonthermal process for bacterial reduction in raw skim milk was examined using a unique pressurized continuous flow system. The lethal effects of subcritical and super-critical CO2 applied at different temperatures and pressures toward total native psychrotrophic microbial populations, total inoculated Pseudomonas fluorescens, and total inoculated spore populations were studied and compared. Pressures between 10.3 and 48.3 MPa; temperatures of 15, 30, 35, and 40 degrees C; and CO2 concentrations of 0, 3, 66, and 132 g/kg of milk were studied. For both native populations and inoculated P. fluorescens, greater total microbial lethality was observed under supercritical CO2 conditions than under subcritical CO2 conditions. At 30 degrees C, there was no effect on total microbial lethality of increasing pressure up to 20.7 MPa with either 66 or 132 g/kg of CO2; at 35 degrees C, there was a positive relationship between pressure and lethality at CO2 levels of 132 g/kg, but no relationship at 66 g/kg of CO2. For total microbial populations and P. fluorescens, CO2 applied at 132 g/kg at 30 degrees C and pressures of 10.3 to 20.7 MPa resulted in an average standard plate count reduction of 3.81 and 2.93 log, respectively; at 35 degrees C and 20.7 MPa, maximum reductions achieved were 5.36 and 5.02 log, respectively. For both total microbial populations and inoculated P. fluorescens, CO2 exhibited a greater overall lethal effect at 132 g/kg than at 66 g/kg and a greater effect at 35 degrees C than at 30 degrees C. At 24.1 and 48.3 MPa and 40 degrees C, microbial lethality in raw aged milk treated with 3 g/kg of CO2 was not significantly different than that observed for uncarbonated milk; lethality achieved in milk treated with 132 g/kg of CO2 was significantly higher than that achieved in these 2 low-level CO2 treatments. No treatment studied had any significant impact on spore populations. Our work shows that, using the studied system, pressurized CO2 results in greater microbial lethality in milk above critical temperatures than below and suggests that a critical concentration threshold level of CO2 is required for lethal effects. Our work also suggests that supercritical CO2 processing in a continuous flow system can achieve reductions in some microbial populations equal to or better than that typically achieved during high-temperature, short-time pasteurization.

The effects of holding raw milk under carbon dioxide pressures of 68 to 689 kPa at temperatures of 5, 6.1, 10, and 20°C on the indigenous microbiota were investigated. These pressure-temperature combinations did not cause precipitation of proteins from the milk. Standard plate counts from treated milks demonstrated significantly lower growth rate compared with untreated controls at all temperatures, and in some cases, the treatment was microcidal. Raw milk treated with CO 2 and held at 6.1°C for 4 d exhibited reduced bacterial growth rates at pressures of 68, 172, 344, and 516 kPa; and at 689 kPa, demonstrated a significant loss of viability in standard plate count assays. The 689-kPa treatment also reduced gram-negative bacteria and total Lactobacillus spp. The time required for raw milk treated at 689 kPa and held at 4°C to reach 4.30 log 10 cfu/mL increased by 4 d compared with untreated controls. Total coliform counts in the treated milk were maintained at 1.95 log 10 cfu/mL by d 9 of treatment, whereas counts in the control significantly increased to 2.61 log 10 cfu/mL by d 4 and 2.89 log 10 cfu/mL by d 9. At d 8, Escherichia coli counts had not significantly changed in treated milk, but significantly increased in the control milk. Thermoduric bacteria counts after 8 d were 1.32 log 10 cfu/mL in treated milk and 1.98 log 10 cfu/mL in control milk. These data indicated that holding raw milk at low CO 2 pressure reduces bacterial growth rates without causing milk protein precipitation. Combining low CO 2 pressure and refrigeration would improve the microbiological quality and safety of raw milk and may be an effective strategy for shipping raw single strength or concentrated milk over long distances.

Inhibition of bacterial growth by dissolved carbon dioxide (CO2) has been well established in many foods including dairy foods. However, the effects of dissolved CO2 on specific growth parameters such as length of lag phase, time to maximum growth rate, and numbers of organisms at the stationary phase have not been quantified for organisms of concern in milk. The effect of dissolved CO2 concentrations of 0.6 to 61.4 mM on specific bacterial growth parameters in raw or single organism inoculated sterile milk was determined at 15 degrees C by conductance. Commingled raw or sterile milks were amended to a final concentration of 0.5 mg/ml each of urea and arginine HCl. Sterile milks were inoculated singly with one of six different microorganisms to a final concentration of approximately 10(2) to 10(3) cfu/ml; raw milk was adjusted to a final indigenous bacterial population of approximately 10(3) cfu/ml. Conductivity of the milk was recorded every 60 s over 4 to 5 d in a circulating apparatus at 15 degrees C. Conductivity values were fit to Gompertz equations and growth parameters calculated. Conductance correlated with plate counts and was satisfactory for monitoring microbial growth. Data fit the Gompertz equation with high correlation (R2 = 0.96 to 1.00). In all cases, dissolved CO2 significantly inhibited growth of raw milk bacteria, influencing lag, exponential, and stationary growth phases as well as all tested monocultures.

The effects of the addition of 11.9 mM CO2 on the growth of Bacillus cereus spores inoculated into sterile homogenized whole milk at 101 and 106 spores/ml and stored at 6.1 degrees C, was examined weekly for 35 d. Colony-forming units from CO2 treated inoculated milks decreased over 35 d at a rate similar to that of untreated inoculated milk, as defined by linear regression. Plate counts for treated and control milks inoculated at 10(1) cfu/ml were not significantly different on sampling d 0, 14, 21, and 28. Plate counts at d 7 were significantly different and counts at d 35 were at undetectable levels for both treated and control milks. Plate counts for milk inoculated at 10(6) cfu/ml were not significantly different on d 0, 28, and 35; they were significantly different on d 7, 14, and 21. There was no consistency as to whether the control or test milks were higher in counts on days when the differences were significant. Added CO2 reduced the pH of the milk from an average value of 6.61 to an average value of 6.31; however, this drop did not correlate with changes in any other parameter measured. These data suggest that moderate levels of CO2 do not enhance the outgrowth of B. cereus spores over long-term storage and do not increase the risk of foodborne illness due to the organism.