Development and optimization of a carbon dioxide-aided cold microfiltration process for the physical removal of microorganisms and somatic cells from skim milk

Physical removal of microorganisms from skim milk by microfiltration (MF) is becoming increasingly attractive to the dairy industry. Typically, this process is performed at temperatures of approximately 50°C. Additional shelf-life and quality benefits might be gained by conducting the MF process at low temperatures. Cold MF could also minimize microbial fouling of the membrane and prevent the germination of thermophilic spores. The objective of this study was to optimize a cold MF process for the effective removal of microbial and somatic cells from skim milk. An experimental MF setup containing a tubular Tami ceramic membrane with a nominal pore size of 1.4 μm was used for MF of raw skim milk at a temperature of 6 ± 1°C. The processing conditions used were cross-flow velocities of 5 to 7 m/s, and transmembrane pressures of 52 to 131 kPa. All MF experiments were performed in triplicate. The permeate flux was determined gravimetrically. Microbiological, chemical, and somatic cell analyses were performed to evaluate the effect of MF on the composition of skim milk. The permeate flux increased drastically when velocity was increased from 5 to 7 m/s. The critical transmembrane pressure range conducive to maximum fluxes was 60 to 85 kPa. When MF was conducted under optimal conditions, very efficient removal of vegetative bacteria, spores, and somatic cells, as well as near complete transmission of proteins into the MF milk, was achieved. To further enhance the flux, a CO₂ backpulsing system was developed. This technique is able both to increase the flux and to maintain it steadily for an extended period of time. The CO₂-aided cold MF process has the potential to become economically attractive to the dairy industry, with direct benefits for the quality and shelf life of dairy products.     

The effect of linear velocity and flux on performance of ceramic graded permeability membranes when processing skim milk at 50°C

Raw milk (about 500 kg) was cold (4°C) separated and then the skim milk was pasteurized at 72°C and a holding time of 16 s. The milk was cooled to 4°C and stored at ≤4°C until processing. The skim milk was microfiltered using a pilot-scale ceramic graded permeability (GP) microfilter system equipped with 0.1-µm nominal pore diameter ceramic Membralox membranes. First, about 155 kg of pasteurized skim milk was flushed through the system to push the water out of the system. Then, additional pasteurized skim milk (about 320 kg) was microfiltered (stage 1) in a continuous feed-and-bleed 3× process using the same membranes. The retentate from stage 1 was diluted with pasteurized reverse osmosis water in a 1:2 ratio and microfiltered (stage 2) with a GP system. This was repeated 3 times, with total of 3 stages in the process (stage 1 = microfiltration; stages 2 and 3 = diafiltration). The results from first 3 stages of the experiment were compared with previous data when processing skim milk at 50°C using ceramic uniform transmembrane pressure (UTP) membranes. Microfiltration of skim milk using ceramic UTP and GP membranes resulted in similar final retentate in terms of serum proteins (SP) removed. The SP removal rate (expressed by kilogram of SP removed per meter-squared of membrane area) was higher for GP membranes for each stage compared with UTP membranes. A higher passage of SP and SP removal rate for GP than UTP membranes was achieved by using a higher cross-flow velocity when processing skim milk. Increasing flux in subsequent stages did not affect membrane permeability and fouling. We operated under conditions that produced partial membrane fouling, due to using a flux that was less than limiting flux but higher than critical flux. Because the critical flux is a function of the cross-flow velocity, the difference in critical flux between UTP and GP membranes resulted only from operating under different cross-flow velocities (6.6 vs 7.12 for UTP and GP membranes, respectively). Conditions that allow microfiltration operation at higher flux will reduce the membrane surface area required to process the same amount of milk in the same length of time. Less membrane surface area reduces investment costs and uses less energy, water, and chemicals to clean the microfiltration system.     

Effect of microfiltration concentration factor on serum protein removal from skim milk using spiral-wound polymeric membranes

Our objective was to determine the effect of concentration factor (CF) on the removal of serum protein (SP) from skim milk during microfiltration (MF) at 50°C using a 0.3-μm-pore-size spiral-wound (SW) polymeric polyvinylidene fluoride (PVDF) membrane. Pasteurized (72°C for 16 s) skim milk was MF (50°C) at 3 CF (1.50, 2.25, and 3.00×), each on a separate day of processing starting with skim milk. Two phases of MF were used at each CF, with an initial startup-stabilization phase (40 min in full recycle mode) to achieve the desired CF, followed by a steady-state phase (90-min feed-and-bleed with recycle) where data was collected. The experiment was replicated 3 times, and SP removal from skim milk was quantified at each CF. System pressures, flow rates, CF, and fluxes were monitored during the 90-min run. Permeate flux increased (12.8, 15.3, and 19.0 kg/m² per hour) with decreasing CF from 3.00 to 1.50×, whereas fouled water flux did not differ among CF, indicating that the effect of membrane fouling on hydraulic resistance of the membrane was similar at all CF. However, the CF used when microfiltering skim milk (50°C) with a 0.3-μm polymeric SW PVDF membrane did affect the percentage of SP removed. As CF increased from 1.50 to 3.00×, the percentage of SP removed from skim milk increased from 10.56 to 35.57%, in a single stage bleed-and-feed MF system. Percentage SP removal from skim milk was lower than the theoretical value. Rejection of SP during MF of skim milk with SW PVDF membranes was caused by fouling of the membrane, not by the membrane itself and differences in the foulant characteristic among CF influenced SP rejection more than it influenced hydraulic resistance. We hypothesize that differences in the conditions near the surface of the membrane and within the pores during the first few minutes of processing, when casein micelles pass through the membrane, influenced the rejection of SP because more pore size narrowing and plugging occurred at low CF than at high CF due to a slower rate of gel layer formation at low CF. It is possible that percentage removal of SP from skim milk at 50°C could be improved by optimization of the membrane pore size, feed solution composition and concentration, and controlling the rate of formation of the concentration polarization-derived gel layer at the surface of the membrane during the first few minutes of processing.     

Pilot-scale crossflow-microfiltration and pasteurization to remove spores of Bacillus anthracis (Sterne) from milk

High-temperature, short-time pasteurization of milk is ineffective against spore-forming bacteria such as Bacillus anthracis (BA), but is lethal to its vegetative cells. Crossflow microfiltration (MF) using ceramic membranes with a pore size of 1.4 μm has been shown to reject most microorganisms from skim milk; and, in combination with pasteurization, has been shown to extend its shelf life. The objectives of this study were to evaluate MF for its efficiency in removing spores of the attenuated Sterne strain of BA from milk; to evaluate the combined efficiency of MF using a 0.8-μm ceramic membrane, followed by pasteurization (72°C, 18.6 s); and to monitor any residual BA in the permeates when stored at temperatures of 4, 10, and 25°C for up to 28 d. In each trial, 95 L of raw skim milk was inoculated with about 6.5 log₁₀ BA spores/mL of milk. It was then microfiltered in total recycle mode at 50°C using ceramic membranes with pore sizes of either 0.8 μm or 1.4 μm, at crossflow velocity of 6.2 m/s and transmembrane pressure of 127.6 kPa, conditions selected to exploit the selectivity of the membrane. Microfiltration using the 0.8-μm membrane removed 5.91 ± 0.05 log₁₀ BA spores/mL of milk and the 1.4-μm membrane removed 4.50 ± 0.35 log₁₀ BA spores/mL of milk. The 0.8-μm membrane showed efficient removal of the native microflora and both membranes showed near complete transmission of the casein proteins. Spore germination was evident in the permeates obtained at 10, 30, and 120 min of MF time (0.8-μm membrane) but when stored at 4 or 10°C, spore levels were decreased to below detection levels (≤0.3 log₁₀ spores/mL) by d 7 or 3 of storage, respectively. Permeates stored at 25°C showed coagulation and were not evaluated further. Pasteurization of the permeate samples immediately after MF resulted in additional spore germination that was related to the length of MF time. Pasteurized permeates obtained at 10 min of MF and stored at 4 or 10°C showed no growth of BA by d 7 and 3, respectively. Pasteurization of permeates obtained at 30 and 120 min of MF resulted in spore germination of up to 2.42 log₁₀ BA spores/mL. Spore levels decreased over the length of the storage period at 4 or 10°C for the samples obtained at 30 min of MF but not for the samples obtained at 120 min of MF. This study confirms that MF using a 0.8-μm membrane before high-temperature, short-time pasteurization may improve the safety and quality of the fluid milk supply; however, the duration of MF should be limited to prevent spore germination following pasteurization.     

Characterization of Proteinaceous Membrane Foulants and Flux Decline During the Early Stages of Whole Milk Ultrafiltration

Pasteurized whole milk was fractionated with a pilot-scale, plate and frame, ultrafiltration system to study membrane fouling and flux decline. Concentration factor was set at approximately 1.4× to simulate the first stage of a multistage UF system. Proteinaceous membrane foulant was characterized by SDS-PAGE. Distribution of proteins in the foulant was very different from distribution of proteins in milk. Whey proteins, α-lactalbumin and β-lactoglobulin, accounted for 95% of the proteinaceous membrane foulants. Very little casein was identified as membrane foulant.The approximate amount of protein in the membrane foulant was estimated to be .6 g/m² of membrane area. Permeate flux studies indicated that flux decline is severe in the early stages of milk ultrafiltration and is associated with irreversible adsorption of protein on the membrane surface. A threefold difference between the water flux of clean membranes and fouled membranes was attributed to the adsorbed foulant. Identification and characterization of membrane foulants and the mechanism of their interaction with membrane surfaces should lead to the design of more efficient ultrafiltration systems for the dairy industry.     

Production efficiency of micellar casein concentrate using polymeric spiral-wound microfiltration membranes

Most current research has focused on using ceramic microfiltration (MF) membranes for micellar casein concentrate production, but little research has focused on the use of polymeric spiral-wound (SW) MF membranes. A method for the production of a serum protein (SP)-reduced micellar casein concentrate using SW MF was compared with a ceramic MF membrane. Pasteurized (79°C, 18s) skim milk (1,100 kg) was microfiltered at 50°C [about 3 × concentration] using a 0.3-μm polyvinylidene fluoride spiral-wound membrane, bleed-and-feed, 3-stage process, using 2 diafiltration stages, where the retentate was diluted 1:2 with reverse osmosis water. Skim milk, permeate, and retentate were analyzed for SP content, and the reduction of SP from skim milk was determined. Theoretically, 68% of the SP content of skim milk can be removed using a single-stage 3× MF. If 2 subsequent water diafiltration stages are used, an additional 22% and 7% of the SP can be removed, respectively, giving a total SP removal of 97%. Removal of SP greater than 95% has been achieved using a 0.1-μm pore size ceramic uniform transmembrane pressure (UTP) MF membrane after a 3-stage MF with diafiltration process. One stage of MF plus 2 stages of diafiltration of 50°C skim milk using a polyvinylidene fluoride polymeric SW 0.3-μm membrane yielded a total SP reduction of only 70.3% (stages 1, 2, and 3: 38.6, 20.8, and 10.9%, respectively). The SP removal rate for the polymeric SW MF membrane was lower in all 3 stages of processing (stages 1, 2, and 3: 0.05, 0.04, and 0.03 kg/m² per hour, respectively) than that of the comparable ceramic UTP MF membrane (stages 1, 2, and 3: 0.30, 0.11, and 0.06 kg/m² per hour, respectively), indicating that SW MF is less efficient at removing SP from 50°C skim milk than the ceramic UTP system. To estimate the number of steps required for the SW system to reach 95% SP removal, the third-stage SP removal rate (27.4% of the starting material SP content) was used to extrapolate that an additional 5 water diafiltration stages would be necessary, for a total of 8 stages, to remove 95% of the SP from skim milk. The 8-plus stages necessary to remove >95% SP for the SW MF membrane would create more permeate and a lengthier process than required with ceramic membranes.     

Clarification of passion fruit juice by microfiltration: Analyses of operating parameters,study of membrane fouling and juice quality

In this study, the performance of two membranes were compared – tubular ceramic and hollow fiber poly(imide) – under transmembrane pressure of 0.5 and 1 bar, for the clarification of passion fruit pulp pre-treated by centrifugation and enzymatic treatment at the concentrations of 150 and 300 ppm. Nutritional and sensorial qualities of the clarified juice obtained were evaluated. Thus, it was possible to observe that the most adequate condition for the clarification of passion fruit pulp was with enzymatic treatment at 150 ppm and its posterior microfiltration at the ceramic tubular membrane of 0.3 μm with transmembrane pressure of 0.5 bar. The fouling mechanism was identified by estimation of model parameters according to a nonlinear regression by Bayesian inference. Analysis of the fouling mechanism results revealed that hollow fiber membrane is controlled by a cake filtration mechanism, and internal pore blocking fouling mechanism controls ceramic tubular membrane.     

Nanofiltration membrane fouling by oppositely charged macromolecules: investigation on flux behavior, foulant mass deposition, and solute rejection

Nanofiltration membrane fouling by oppositely charged polysaccharide (alginate) and protein (lysozyme) was systematically studied. It was found that membrane flux decline in the presence of both lysozyme and alginate was much more severe compared to that when there was only lysozyme or alginate in the feed solution. The flux performance for the mixed foulants was only weakly affected by solution pH and calcium concentration. These effects were likely due to the strong electrostatic attraction between the two oppositely charged foulants. Higher initial flux caused increased foulant deposition, more compact foulant layer, and more severe flux decline. The deposited foulant cake layer had a strong tendency to maintain a constant foulant composition that was independent of the membrane initial flux and only weakly dependent on the relative foulant concentration in feed solution. In contrast, solution chemistry (pH and [Ca2?]) had marked effect on the foulant layer composition, likely due to the resulting changes in the foulant-foulant interaction. The mixed alginate-lysozyme fouling could result in an initial enhancement in salt rejection. However, such initial enhancement was not observed when there was 1 mM calcium present in the feedwater, which may be attributed to the charge neutralization of the foulant layer.     

Shelf life of pasteurized microfiltered milk containing 2% fat

The goal of this research was to produce homogenized milk containing 2% fat with a refrigerated shelf life of 60 to 90 d using minimum high temperature, short time (HTST) pasteurization in combination with other nonthermal processes. Raw skim milk was microfiltered (MF) using a Tetra Alcross MFS-7 pilot plant (Tetra Pak International SA, Pully, Switzerland) equipped with Membralox ceramic membranes (1.4 μm and surface area of 2.31 m²; Pall Corp., East Hills, NY). The unpasteurized MF skim permeate and each of 3 different cream sources were blended together to achieve three 2% fat milks. Each milk was homogenized (first stage: 17 MPa, second stage: 3 MPa) and HTST pasteurized (73.8°C for 15 s). The pasteurized MF skim permeate and the 3 pasteurized homogenized 2% fat milks (made from different fat sources) were stored at 1.7 and 5.7°C and the standard plate count for each milk was determined weekly over 90 d. When the standard plate count was >20,000 cfu/mL, it was considered the end of shelf life for the purpose of this study. Across 4 replicates, a 4.13 log reduction in bacteria was achieved by MF, and a further 0.53 log reduction was achieved by the combination of MF with HTST pasteurization (73.8°C for 15 s), resulting in a 4.66 log reduction in bacteria for the combined process. No containers of MF skim milk that was pasteurized after MF exceeded 20,000 cfu/mL bacteria count during 90 d of storage at 5.7°C. The 3 different approaches used to reduce the initial bacteria and spore count of each cream source used to make the 2% fat milks did not produce any shelf-life advantage over using cold separated raw cream when starting with excellent quality raw whole milk (i.e., low bacteria count). The combination of MF with HTST pasteurization (73.8°C for 15 s), combined with filling and packaging that was protected from microbial contamination, achieved a refrigerated shelf life of 60 to 90 d at both 1.7 and 5.7°C for 2% fat milks.     

Shelf life and quality of skim milk processed by cold microfiltration with a 1.4-μm pore size membrane,with or without heat treatment

The objective of this study was to evaluate the effectiveness of cold microfiltration (MF), alone or in combination with heat treatment, in extending the shelf life of skim milk. Raw skim milk underwent MF at 6 ± 1°C with a ceramic membrane of 1.4-μm pore size, at a transmembrane pressure of 75.8 kPa and a crossflow velocity of 7 m/s. Samples of raw skim milk; MF skim milk; high-temperature, short-time (HTST)-pasteurized milk; and MF+HTST-pasteurized skim milk were stored at 6°C for 92 d. During the shelf-life study, the total bacterial count and degree of proteolysis were evaluated weekly. The study was replicated 3 times. Cold MF was very effective in reducing the microbial load in skim milk, and an average of 3.4 log reduction in vegetative bacteria was obtained. Although HTST pasteurization reduced the bacterial load by ～2 log, the MF+HTST process resulted in near complete elimination of vegetative microflora, with a total of ～4 log reduction. A 9-member sensory panel found no significant differences between skim milk samples processed with or without MF. The MF+HTST skim milk had only minor microbial growth after 92 d at 6°C, but its proteolytic shelf life was limited by plasmin activity. A reduction of plasmin activity and a slower rate of proteolysis were obtained by increasing the heat treatment temperature to 85°C. The results of this study can be used to make decisions regarding processing strategies that lead to increased skim milk shelf life.     

Performance assessment of membrane distillation for skim milk and whey processing

Membrane distillation is an emerging membrane process based on evaporation of a volatile solvent. One of its often stated advantages is the low flux sensitivity toward concentration of the processed fluid, in contrast to reverse osmosis. In the present paper, we looked at 2 high-solids applications of the dairy industry: skim milk and whey. Performance was assessed under various hydrodynamic conditions to investigate the feasibility of fouling mitigation by changing the operating parameters and to compare performance to widespread membrane filtration processes. Whereas filtration processes are hydraulic pressure driven, membrane distillation uses vapor pressure from heat to drive separation and, therefore, operating parameters have a different bearing on the process. Experimental and calculated results identified factors influencing heat and mass transfer under various operating conditions using polytetrafluoroethylene flat-sheet membranes. Linear velocity was found to influence performance during skim milk processing but not during whey processing. Lower feed and higher permeate temperature was found to reduce fouling in the processing of both dairy solutions. Concentration of skim milk and whey by membrane distillation has potential, as it showed high rejection (>99%) of all dairy components and can operate using low electrical energy and pressures (<10 kPa). At higher cross-flow velocities (around 0.141 m/s), fluxes were comparable to those found with reverse osmosis, achieving a sustainable flux of approximately 12 kg/h·m² for skim milk of 20% dry matter concentration and approximately 20 kg/h·m² after 18 h of operation with whey at 20% dry matter concentration.     

Analysis of the fouling mechanisms during cross-flow ultrafiltration of apple juice

The purpose of this work was to study the fouling mechanisms of a Carbosep^® M8 membrane during the cross-flow ultrafiltration of apple juice. A new fouling model has been developed that simultaneously considers membrane blocking within the pores, at the pore mouths and by cake formation at the membrane surface. Membrane fouling by apple juice was due to internal pore blocking as well as cake formation. When operating ultrafiltration at a transmembrane pressure of 150 kPa and a cross-flow velocity of 7 m/s, fouling was minimal with a gradual decrease of the relative contribution of cake formation; however, transmembrane pressure still exceeds critical pressure. The fouling model predicts no cake formation at a cross-flow velocity of 7.4 m/s and a transmembrane pressure of 150 kPa or at a cross-flow velocity of 7.0 m/s and a transmembrane pressure of 120 kPa. Under these conditions, internal membrane blocking would be the only mechanism responsible for the decrease of permeate flux.     

Pilot-scale ceramic membrane filtration of skim milk for the production of a protein base ingredient for use in infant milk formula

The protein composition of bovine skim milk was modified using pilot scale membrane filtration to produce a whey protein-dominant ingredient with a casein profile closer to human milk. Bovine skim milk was processed at low (8.9 °C) or high (50 °C) temperature using ceramic microfiltration (MF) membranes (0.1 μm mean pore diameter). The resulting permeate stream was concentrated using polyethersulfone ultrafiltration (UF) membranes (10 kDa cut-off). The protein profile of MF and UF retentate streams were determined using reversed phase-high performance liquid chromatography and polyacrylamide gel electrophoresis. Permeate from the cold MF process (8.9 °C) had a casein:whey protein ratio of ∼35:65 with no α_S- or κ-casein present, compared with a casein:whey protein ratio of ∼10:90 at 50 °C. This study has demonstrated the application of cold membrane filtration (8.9 °C) at pilot scale to produce a dairy ingredient with a protein profile closer to that of human milk.     

Effect of skim milk treated with high hydrostatic pressure on permeate flux and fouling during ultrafiltration

Ultrafiltration (UF) is largely used in the dairy industry to generate milk and whey protein concentrate for standardization of milk or production of dairy ingredients. Recently, it was demonstrated that high hydrostatic pressure (HHP) extended the shelf life of milk and improved rennet coagulation and cheese yield. Pressurization also modified casein micelle size distribution and promoted aggregation of whey proteins. These changes are likely to affect UF performance. Consequently, this study determined the effect of skim milk pressurization (300 and 600 MPa, 5 min) on UF performance in terms of permeate flux decline and fouling. The effect of HHP on milk proteins was first studied and UF was performed in total recycle mode at different transmembrane pressures to determine optimal UF operational parameters and to evaluate the effect of pressurization on critical and limiting fluxes. Ultrafiltration was also performed in concentration mode at a transmembrane pressure of 345 kPa for 130 or 140 min to evaluate the decline of permeate flux and to determine fouling resistances. It was observed that average casein micelle size decreased by 32 and 38%, whereas β-lactoglobulin denaturation reached 30 and 70% at 300 and 600 MPa, respectively. These results were directly related to UF performance because initial permeate fluxes in total recycle mode decreased by 25% at 300 and 600 MPa compared with nonpressurized milk, critical flux, and limiting flux, which were lower during UF of milk treated with HHP. During UF in concentration mode, initial permeate fluxes were 30% lower at 300 and 600 MPa compared with the control, but the total flux decline was higher for nonpressurized milk (62%) compared with pressure-treated milk (30%). Fouling resistances were similar, whatever the treatment, except at 600 MPa where irreversible fouling was higher. Characterization of the fouling layer showed that caseins and β-lactoglobulin were mainly involved in membrane fouling after UF of pressure-treated milk. Our results demonstrate that HHP treatment of skim milk drastically decreased UF performance.     

Efficiency of serum protein removal from skim milk with ceramic and polymeric membranes at 50°C

Raw milk (2,710 kg) was separated at 4°C, the skim milk was pasteurized (72°C, 16 s), split into 3 batches, and microfiltered using pilot-scale ceramic uniform transmembrane pressure (UTP; Membralox model EP1940GL0.1μA, 0.1 μm alumina, Pall Corp., East Hills, NY), ceramic graded permeability (GP; Membralox model EP1940GL0.1μAGP1020, 0.1 μm alumina, Pall Corp.), and polymeric spiral-wound (SW; model FG7838-OS0x-S, 0.3 μm polyvinylidene fluoride, Parker-Hannifin, Process Advanced Filtration Division, Tell City, IN) membranes. There were differences in flux among ceramic UTP, ceramic GP, and polymeric SW microfiltration membranes (54.08, 71.79, and 16.21 kg/m² per hour, respectively) when processing skim milk at 50°C in a continuous bleed-and-feed 3× process. These differences in flux among the membranes would influence the amount of membrane surface area required to process a given volume of milk in a given time. Further work is needed to determine if these differences in flux are maintained over longer processing times. The true protein contents of the microfiltration permeates from UTP and GP membranes were higher than from SW membranes (0.57, 0.56, and 0.38%, respectively). Sodium-dodecyl-sulfate-PAGE gels for permeates revealed a higher casein proportion in GP and SW permeate than in UTP permeate, with the highest passage of casein through the GP membrane under the operational conditions used in this study. The slight cloudiness of the permeates produced using the GP and SW systems may have been due to the presence of a small amount of casein, which may present an obstacle in their use in applications when clarity is an important functional characteristic. More β-lactoglobulin passed through the ceramic membranes than through the polymeric membrane. The efficiency of removal of serum proteins in a continuous bleed-and-feed 3× process at 50°C was 64.40% for UTP, 61.04% for GP, and 38.62% for SW microfiltration membranes. The SW polymeric membranes had a much higher rejection of serum proteins than did the ceramic membranes, consistent with the sodium-dodecyl-sulfate PAGE data. Multiple stages and diafiltration would be required to produce a 60 to 65% serum protein reduced micellar casein concentrate with SW membranes, whereas only one stage would be needed for the ceramic membranes used in this study.     

A comparison between ceramic and polymeric membrane systems for casein concentrate manufacture

Skim milk microfiltration (MF) is a well established dairy processing operation to remove whey proteins. This is classically completed using ceramic membranes under the uniform transmembrane pressure (UTMP) concept. However, there is a growing interest in the use of spiral wound polymeric membranes for this purpose due to their lower capital cost. In this work we show that polymeric and ceramic membranes provide comparable performance during skim milk MF when operated at comparable levels of transmembrane pressure (TMP), shear stress and temperature. Operation is strongly affected by both TMP and temperature but is less affected by crossflow velocity under the conditions utilised here. However, while polymeric membranes offer similar performance, it will be operationally difficult to retain the necessarily low TMP at the industrial scale, due to the lack of a comparable UTMP configuration.     

Efficient removal of spores from skim milk using cold microfiltration: Spore size and surface property considerations

Bacterial spores present in milk can cause quality and shelf-life issues for dairy products. The objectives of this study were to evaluate the effectiveness of microfiltration (MF) in removing Bacillus licheniformis and Geobacillus sp. spores from skim milk using membranes with pore sizes of 1.4 and 1.2 µm, and to investigate the role of spore surface properties in MF removal. Cell sizes were determined by scanning electron microscopy, surface charge by zeta potential analysis, and surface hydrophobicity by contact angle measurements. Commercially pasteurized skim milk was inoculated with a spore suspension at about 10⁶ cfu/mL, and then processed by MF using ceramic membranes at 6°C, a cross-flow velocity of 4.1 m/s, and transmembrane pressure of 69 to 74 kPa. Total aerobic plate and spore counts in the milk were determined before and after MF. All processing runs and surface and product analyses were conducted in triplicate, and data were analyzed statistically. For the same strain, spores were shorter and wider than vegetative cells, averaging 1.37 to 1.59 µm in length and 0.64 to 0.81 µm in width. Reduction of B. licheniformis spores significantly increased with a reduction in MF pore size, from 2.17 log for 1.4-µm pore size, to 4.57 log for 1.2-µm pore size. Both pore sizes resulted in almost complete removal of Geobacillus sp. spores. All spores and the ceramic membrane had a negative surface charge at milk pH, indicating an electrostatic repulsion between them. Bacillus licheniformis spores were hydrophilic, whereas Geobacillus sp. spores were hydrophobic. Consequently, Geobacillus sp. spores had a tendency to cluster in skim milk, preventing their passage even through the 1.4-µm MF membrane, whereas some B. licheniformis spores could still pass through a 1.2-µm membrane. This study demonstrates that efficient removal of spores from skim milk by cold MF may require a smaller membrane pore size than required for removal of vegetative cells of the same species, and that cell surface properties may interfere with the outcome of filtration as would be anticipated based on size alone.     

Microbial inactivation and shelf life comparison of 'cold' hurdle processing with pulsed electric fields and microfiltration, and conventional thermal pasteurisation in skim milk

Thermal pasteurisation (TP) is the established food technology for commercial processing of milk. However, degradation of valuable nutrients in milk and its sensory characteristics occurs during TP due to substantial heat exposure. Pulsed electric fields (PEF) and microfiltration (MF) both represent emerging food processing technologies allowing gentle milk preservation at lower temperatures and shorter treatment times for similar, or better, microbial inactivation and shelf stability when applied in a hurdle approach compared to TP. Incubated raw milk was used as an inoculum for the enrichment of skim milk with native microorganisms before PEF, MF, and TP processing. Inoculated milk was PEF-processed at electric field strengths between 16 and 42 kV/cm for treatment times from 612 to 2105 μs; accounting for energy densities between 407 and 815 kJ/L, while MF was applied with a transmembrane flux of 660 L/h m². Milk was TP-treated at 75 °C for 24 s. Comparing PEF, MF, and TP for the reduction of the native microbial load in milk led to a 4.6 log₁₀ CFU/mL reduction in count for TP, which was similar to 3.7 log₁₀ CFU/mL obtained by MF (P ≥ 0.05), and more effective than the 2.5 log₁₀ CFU/mL inactivation achieved by PEF inactivation (at 815 kJ/L (P < 0.05)). Combined processing with MF followed by PEF (MF/PEF) produced a 4.1 (at 407 and 632 kJ/L), 4.4 (at 668 kJ/L) and 4.8 (at 815 kJ/L) log₁₀ CFU/mL reduction in count of the milk microorganisms, which was comparable to that of TP (P ≥ 0.05). Reversed processing (PEF/MF) achieved comparable reductions of 4.9, 5.3 and 5.7 log₁₀ CFU/mL (at 407, 632 and 668 kJ/L, respectively (P ≥ 0.05)) and a higher inactivation of 7.1 log₁₀ (at 815 kJ/mL (P < 0.05)) in milk than for TP. Microbial shelf life of PEF/MF-treated (815 kJ/L) and TP-treated milk stored at 4 °C was analysed over 35 days for total aerobic; enterobacteria; yeasts and moulds; lactobacilli; psychrotroph; thermoduric psychrotroph, mesophilic, and thermophilic; and staphylococci counts. For both PEF/MF and TP-treated milk an overall shelf stability of 7 days was observed based on total aerobic counts (P ≥ 0.05). Milk hurdle processing with PEF/MF at its most effective treatment parameters produced greater microbial inactivation and overall similar shelf stability at lower processing temperatures compared to TP. With higher field strength, shorter treatment time, larger energy density, and rising temperature the efficacy of PEF/MF increased contrary to MF/PEF. Thus, PEF/MF represents a potential alternative for ‘cold’ pasteurisation of milk with improved quality.     

Gelation of casein micelles in ��-casein reduced milk prepared using membrane filtration

Pasteurized skim milk was subjected to membrane filtration using a molecular weight cut-off of 80 kDa and a plate and frame pilot scale system at temperatures below 10 °C. Via this process, transmission of whey proteins and ??-casein through the membrane was achieved. The milk was concentrated to two times (based on volume reduction), and whey protein-free permeate was added to return to the original volume fraction of casein micelles in milk. This diafiltration process was carried out four times, and the retentate obtained was nearly free of whey proteins and with approximately 20% of ??-casein removed. The same membrane filtration was also carried out at 25 °C to achieve transmission of whey protein but not of ??-casein, and to obtain whey protein-depleted milk without depletion of ??-casein.The gelling behaviour of these samples, reconstituted to the original casein volume fraction, was examined using rheology and diffusing wave spectroscopy. When compared to the original skim milk it was found that there were no statistically significant differences in gelation behaviour during acidification, but differences were noted in gelation time and final stiffness modulus for samples undergoing renneting. These differences were attributed mostly to the changes in ionic composition, as when the serum composition of the retentates was re-equilibrated against the original skim milk by dialysis; the gelation behaviour of the samples was comparable to that of skim milk. The results clearly indicate the importance of the milk's overall ionic balance in the early stages of aggregation of rennet-induced gelation of milk.     

Processing factors that influence casein and serum protein separation by microfiltration

Our objective was to demonstrate the effect of various processing factors on the performance of a microfiltration system designed to process skim milk and separate casein (CN) from serum proteins (SP). A mathematical model of a skim milk microfiltration process was developed with 3 stages plus an additional fourth finishing stage to standardize the retentate to 9% true protein (TP) and allow calculation of yield of a liquid 9% TP micellar CN concentrate (MCC) and milk SP isolate (MSPI; 90% SP on a dry basis). The model was used to predict the effect of 5 factors: 1) skim milk composition, 2) heat treatment of skim milk, 3) concentration factor (CF) and diafiltration factor (DF), 4) control of CF and DF, and 5) SP rejection by the membrane on retentate and permeate composition, SP removal, and MCC and MSPI yield. When skim milk TP concentration increased from 3.2 to 3.8%, the TP concentration in the third stage retentate increased from 7.92 to 9.40%, the yield of MCC from 1,000 kg of skim milk increased from 293 to 348 kg, and the yield of MSPI increased from 6.24 to 7.38 kg. Increased heat treatment (72.9 to 85.2°C) of skim milk caused the apparent CN as a percentage of TP content of skim milk as measured by Kjeldahl analysis to increase from 81.97 to 85.94% and the yield of MSPI decreased from 6.24 to 4.86 kg, whereas the third stage cumulative percentage SP removal decreased from 96.96 to 70.08%. A CF and DF of 2× gave a third stage retentate TP concentration of 5.38% compared with 13.13% for a CF and DF of 5×, with the third stage cumulative SP removal increasing from 88.66 to 99.47%. Variation in control of the balance between CF and DF (instead of an equal CF and DF) caused either a progressive increase or decrease in TP concentration in the retentate across stages depending on whether CF was greater than DF (increasing TP in retentate) or CF was less than DF (decreasing TP in retentate). An increased rejection of SP by the membrane from an SP removal factor of 1 to 0.6 caused a reduction in MSPI yield from 6.24 to 5.19 kg/1,000 kg of skim milk, and third stage cumulative SP removal decreased from 96.96 to 79.74%. Within the ranges of the 5 factors studied, the TP content of the third stage retentate was most strongly affected by the target CF and DF and variation in skim milk composition. Cumulative percentage SP removal was most strongly affected by the heat treatment of skim milk, the SP removal factor, and the target CF and DF. The MCC yield was most strongly affected by initial skim milk composition. Yield of MSPI was strongly affected by skim milk composition, whereas the heat treatment of milk and SP removal factor also had a large effect.