Pulsed electric field pre-treatment for enhanced bacterial survival after drying: Effect of carrier matrix and strain variability

Previous studies suggest that increased intracellular trehalose concentrations can result in increased robustness of probiotics and starter cultures after processing. We evaluated whether pulsed electric field (PEF) pre-treatment, resulting in increased intracellular trehalose concentrations, can lead to enhanced survival during spray drying and subsequent storage and what the effect of the carrier matrix during drying herein is on survival. The applied PEF pre-treatment resulted in increased survival of Lactobacillus plantarum WCFS1 after spray drying and subsequent storage in reconstituted skim milk (RSM), though not in the other evaluated carrier matrices. The same PEF pre-treatment in lactose instead of trehalose electroporation medium resulted also in an increased survival after spray drying in RSM. Further analysis of intracellular trehalose and lactose concentrations revealed that survival after spray drying cannot only be explained by intracellular trehalose and/or lactose concentrations. Experiments with other bacterial strains indicated that strain variability cannot be neglected when designing a process aimed at enhanced bacterial survival after processing. Overall, PEF pre-treatment is promising for enhancing survival of bacteria after drying and storage, though further understanding of the applicability is required for industrial application.Industrial relevance textProbiotics and starter cultures are very common in food industry. Survival of these cultures during drying processes is essential for their application. A pulsed electric field pre-treatment resulting in increased bacterial survival after spray drying and powder storage may contribute to more energy efficient production processes of dried bacterial cultures.     

The effect of heat treatment on intrinsic and fortified selenium levels in cow's milk

The effects of two heat processing methods (pasteurisation and spray drying) routinely used in the processing of cow's milk and the production of infant formula powder on the selenium (Se) content of liquid milk, milk fortified with sodium selenite and sodium selenate were studied. Pasteurisation reduced intrinsic Se and selenate levels by 7.9% and 6.2% at p < 0.05 level and selenite levels by 7.0% at p > 0.05 level. Se losses following spray drying were 44.8% (p < 0.001), 11.4% (p < 0.01) and 10.0% (p < 0.01) for intrinsic selenium, selenite and selenate fortified milk, respectively. Total Se losses from unprocessed milk following processing (pasteurisation and spray drying) were 49.2% (p < 0.001), 17.6% (p < 0.001) and 15.6% (p < 0.001) for intrinsic selenium, selenite and selenate fortified milk, respectively.     

Proteomic study on the stability of proteins in bovine,camel, and caprine milk sera after processing

Milk proteins have been shown to be very sensitive to processing. This study aims to investigate the changes of the bovine, camel, and caprine milk proteins after freezing, pasteurization (62 °C, 30 min), and spray drying by proteomic techniques, filter-aided sample preparation (FASP) and dimethyl labeling followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). A total of 129, 125, and 74 proteins were quantified in bovine, camel, and caprine milk sera, respectively. The milk serum protein content decreased significantly after freezing, pasteurization, or spray drying, which can be ascribed to the removal of protein aggregates by the pH adjustment and ultracentrifugation during sample preparation. Some of the immune-related proteins were heat-sensitive, such as lactoferrin (LTF), glycosylation-dependent cell adhesion molecule 1 (GLYCAM1), and lactadherin (MFGE8), with losses of approximately 25% to 85% after pasteurization and 85% to 95% after spray drying. α-Lactalbumin (LALBA), osteopontin (SPP1), and whey acidic protein (WAP) were relatively heat stable with losses of 10% to 50% after pasteurization and 25% to 85% after spray drying. The increase of some individual proteins in concentration after freezing may be caused by the proteins originating from damaged milk fat globules and somatic cells. GLYCAM1 decreased significantly after pasteurization in bovine and camel milk but this protein is relatively stable in caprine milk. In conclusion, milk proteins changed differently in concentration after different processing steps, as well as among different species.     

Comparative study on the microstructure and functional properties of casein in goat milk processed by different methods

This work compared the thermal stability, microstructure and functional properties of casein (CN) in goat milk processed by different methods, including homogenisation, low-temperature long-time (LTLT), high-temperature short-time (HTST), ultrapasteurisation (UP) and ultrahigh temperature (UHT) treatment. The greatest development of CN-whey protein complexes was observed in LTLT goat milk and the least in HTST goat milk. The CN micelles in LTLT, UP and UHT milk were more obviously changed than the CN micelles in HTST milk, and the random coil content increased significantly after the homogenisation, UP and UHT treatments. The homogenisation had a significant effect on the functional properties of CN. In addition, the most obvious changes in the properties of CN were observed in LTLT and UHT goat milk. The results suggested that homogenisation, pasteurisation and UHT can cause effective changes in the functional properties of CN in goat milk that facilitate its application in the milk industry.     

Physical properties of goat milk powder with soy lecithin added before spray drying

In this study, physical characteristics of goat milk powder produced with the addition of soy lecithin at levels of 0 (control), 0.4, 0.8 and 1.0 g lecithin/100 g of total solids in concentrated milk before the spray drying process were investigated. Goat milk was pasteurised, concentrated at 40% of total solids, spray dried and packed in plastic bags under vaccum conditions. Lecithin addition decreased the wetting time of milk powders, although no influence was observed on dispersibility, water sorption, water activity and particle size distribution of the powders. Powders with higher levels of lecithin showed significantly lower brightness, with a greater intensity of yellow colour. It was concluded that lecithin addition before spray drying process at the minimal proportion in concentrated milk of 0.4 g lecithin/100 g of total solids in concentrated milk is useful for achieving more rapid wetting time of goat milk powder.     

The significance of critical processing steps in the production of dried egg albumen powder on gel textural and foaming properties

The purpose of the present work was to study the variation in egg albumen functional properties as a function of seven selected processing steps from the initially raw liquid albumen to the final dried egg albumen powder. Albumen samples in six replicates were analysed for dry matter, pH, glucose, triglycerides, albumen gel properties (ie textural stress and Hencky strain, water‐ and protein‐binding capacity and gel colour L*, a*, b*), foaming properties (ie overrun, stability against drainage and bubble breakdown) and surface tension. The gel texture and water‐holding capacity significantly decreased during the processing steps from raw albumen through storage, centrifugation and ion exchange, whereas the final dry‐pasteurisation resulted in increased gel properties. Covalently linked protein polymers formed during the dry‐pasteurisation, as revealed by SDS‐PAGE, may explain this improvement. The foam overrun increased twofold during the three final steps of ultrafiltration, spray‐drying and dry‐pasteurisation compared with the raw albumen; however, the foam stability decreased, ie drainage and foam volume breakdown rates increased. The surface pressure increase was positively correlated with the foam overrun. This paper reveals at which processing steps the control of functional properties of egg albumen powder can take place. Copyright © 2004 Society of Chemical Industry     

Computational fluid dynamics studies on pasteurisation of canned milk

Pasteurisation is a vital unit operation to inactivate the spoilage organisms and enzymes present in the milk. Computational fluid dynamics (CFD) modelling can be used as a tool to determine the temperature distribution pattern inside canned milk during thermal processing. In this study, a CFD model was developed for the pasteurisation process of canned milk at 74°C and 85°C and validated with the experimental measurements. Moreover, the effect of can rotation (5, 50 and 100 rpm) on processing time and pasteurisation value was investigated. This study predicted rotation of can with 5 rpm was more effective than stationary positioned can during pasteurisation.     

Behaviour of homogenized fat globules during the spray drying of whole milk

The changes in milk fat globule size and fat globule surface proteins of both low-preheated and high-preheated concentrated milks, which were homogenized at low or high pressure prior to spray drying using a disc atomization drier, were examined. The average fat globule size (d₃₂) of the spray-dried milk powders was smaller than that of the corresponding concentrates, but a small proportion of very large globules (4–80 μm) was also formed during spray drying. As a consequence, total surface protein (mg protein g⁻¹ fat) increased due to the adsorption of casein micelles at the fat globule surface during spray drying. Confocal micrographs of the powders showed some apparent spreading of the fat on the surface of the powder particles, particularly when the concentrates were homogenized at low pressure. These results indicate disruption of the milk fat globules during spray drying, which consequently causes changes in the fat globule surface protein layer.     

枯草芽孢杆菌Prob1822复合抗热保护剂的研究

以枯草芽孢杆菌（Bacillus subtilis）Prob1822为研究对象,以热诱激处理（55 ℃热处理10 min）稳定期菌体存活率为评价指标,通过单因素试验和响应面法研究枯草芽孢杆菌Prob1822复合抗热保护剂的组成。结果表明,单一抗热保护剂以海藻糖、蔗糖及脱脂奶粉抗热保护效果较好;利用响应面法优化复合抗热保护剂配方为海藻糖9.0%、蔗糖5.0%和脱脂奶粉6.8%。在此最优条件下,菌体存活率为（95.24±0.84）%。糖类与蛋白质联用作为复合抗热保护剂比单一保护剂抗热保护效果更好,可减轻喷雾干燥对菌体亚细胞结构及生物大分子损伤,实现枯草芽孢杆菌的抗热保护。     

海藻糖对纤维素酶的干燥保护作用

纤维素酶液中加入不同剂量的海藻糖作为干燥保护剂,喷雾干燥后纤维素酶活力的回收率由空白对照的63.7%提高到89.0%;硅藻土吸附酶液经50℃干燥后,空白对照的酶活力回收率为60.5%,海藻糖组的回收率为65.8%;90℃10min造粒后,纤维素酶活力的回收率由空白对照的62.4%提高到92.7%。造粒的纤维素酶置50℃保存240h后,空白对照的剩余酶活力为70.5%,而海藻糖组的酶活力仍保持在81.2%。结果表明,海藻糖在纤维素酶的干燥和保存过程中有较好的保护效果。     

牛羊乳酪蛋白制备及电泳分析比较

采采用新鲜和复原的牛羊乳为原料,等电点沉淀,通过洗涤、干燥等步骤分别制得牛乳和羊乳酪蛋白,用凯氏定氮法对其进行定量检测,应用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS—PAGE）分析比较牛羊乳酪蛋白组分差异。结果表明：牛乳酪蛋白得率为86．75％,羊乳酪蛋白得率为90．55％,高含量的酪蛋白主要集中在电泳图谱的中分子量组,可分为ds,CN、as2-CN、β-CN和K—CN,牛羊乳as2-CN分子量羊乳大于牛乳,牛乳a-CN含量比较多,羊乳β—CN含量比较多,鲜乳与复原乳全蛋白主要组分在电泳图谱中除酪蛋白差别外,上端的高分子量组清蛋白区羊乳IgG重链的分子量比牛乳小。应用蛋白质电泳分析技术可以区分羊乳和牛乳的蛋白质组分,等电点沉淀法制备酪蛋白的方法简单,易于操作。     

Effects of polymerized goat milk whey protein on physicochemical properties and microstructure of recombined goat milk yogurt

Goat milk whey protein concentrates were manufactured by microfiltration (MF) and ultrafiltration (UF). When MF retentate blended with cream, which could be used as a starting material in yogurt making. The objective of this study was to prepare goat milk whey protein concentrates by membrane separation technology and to investigate the effects of polymerized goat milk whey protein (PGWP) on the physicochemical properties and microstructure of recombined goat milk yogurt. A 3-stage MF study was conducted to separate whey protein from casein in skim milk with 0.1-µm ceramic membrane. The MF permeate was ultrafiltered using a 10 kDa cut-off membrane to 10-fold, followed by 3 step diafiltration. The ultrafiltration-diafiltration-treated whey was electrodialyzed to remove 85% of salt, and to obtain goat milk whey protein concentrates with 80.99% protein content (wt/wt, dry basis). Recombined goat milk yogurt was prepared by mixing cream and MF retentate, and PGWP was used as main thickening agent. Compared with the recombined goat milk yogurt without PGWP, the yogurt with 0.50% PGWP had desirable viscosity and low level of syneresis. There was no significant difference in chemical composition and pH between the recombined goat milk yogurt with PGWP and control (without PGWP). Viscosity of all the yogurt samples decreased during the study. There was a slight but not significant decrease in pH during storage. Bifidobacterium and Lactobacillus acidophilus in yogurt samples remained above 10⁶ cfu/g during 8-wk storage. Scanning electron microscopy of the recombined goat milk yogurt with PGWP displayed a compact protein network. Results indicated that PGWP prepared directly from raw milk may be a novel protein-based thickening agent for authentic goat milk yogurt making.     

Spray‐drying encapsulation of citral in sucrose or trehalose matrices: physicochemical and sensory characteristics

The potential of the disaccharide trehalose as carrier for producing spray‐dried citral was examined. Some physical and sensory characteristics of citral encapsulated in matrices containing either trehalose or sucrose and maltodextrin (MD) were analysed. They included water sorption, glass transition temperatures (T_g), nuclear magnetic resonance (NMR) relaxation and citral retention during spray‐drying. The glassy state at room temperature (25 °C) was maintained as far as 33% relative humidities (RH) for the spray‐dried trehalose formulation and 43% RH for trehalose–MD; however, for sucrose–MD and for sucrose formulations, the glassy state was limited to RHs lower than 33% and 22%, respectively. Sensory evaluation and gas chromatographic analysis indicated that citral retention after spray‐drying was similar for matrices containing either trehalose or sucrose. However, trehalose formulations had improved physical stability as compared to sucrose.     

Exploring the protective effects of calcium-containing carrier against drying-induced cellular injuries of probiotics using single droplet drying technique

Protective carriers that encapsulate probiotics in spray drying could improve the survival ratio of dried cells through different mechanisms. Unveiling the protective mechanism of each carrier will contribute to a rational design of high performance carrier formulation. This study utilized single droplet drying (SDD) technique to investigate the effects of calcium cation in varied carrier formulation. Inactivation histories of Lactobacillus rhamnosus GG (LGG) in different carriers were compared, and cellular injury history of probiotics during droplet drying was studied for the first time. Adding 1 mM CaCl₂ to lactose carrier protected cell viability, mitigated cellular injuries, and enhanced regrowth capability as drying progressed, demonstrating the positive effect of Ca^2 + with possible mechanism of stabilizing sub-cellular structures. At later drying stages, cell survival in Lac/Ca carrier was increased by 0.5–1.5 log on selective media compared to lactose carrier. Supplementing calcium-binding agents lowered the protective effect, shortening the initiation of rapid cell inactivation down to 120 s of drying. Adding CaCl₂ to trehalose carrier barely improved cell survival, indicating that the protective effect could be influenced by carrier formulation. Pure trehalose carrier exerted excellent protection on LGG, supporting cells to regrow in liquid rich medium even after 180 s of drying. The protection of trehalose may stem from stabilization of sub-cellular structures, which possibly overlap the effect of Ca^2 +. The findings suggested that high performance carrier formulation might be developed by combining carrier materials with different protective mechanisms, for maximizing the survival of active dry probiotics in industrial spray drying operation.     

Proteomics and microstructure profiling of goat milk protein after homogenization

This study investigated the protein changes in goat milk during the homogenization process using label-free quantification. We quantified 310 and 315 proteins in the control group (CG) and homogenized group (HG), respectively, and 16 proteins were significantly different between the 2 groups. For HG, the goat milk protein particle sizes were smaller and more evenly distributed and exhibited an increase in the regular arrangement of the secondary structures. Proteomics analysis verified that xanthine dehydrogenase and asparaginase-like 1 expression in CG were higher than in HG, whereas the opposite was observed for fructose-bisphosphate aldolase, κ-casein, and β-casein. Significant changes were found in the homogenization-treated goat milk proteome that were related to goat milk glycolysis/gluconeogenesis metabolism. This work provides updated information on the current proteome characteristics of homogenized goat milk, which may be important for applying the protein component of goat milk to human nutrition and health.     

Effect of extended storage on microbiological quality, somatic cell count, and composition of raw goat milk on a farm

Dairy goat herds in the United States generally are small, widely scattered, and distant from processing facilities. Unlike the situation for cow milk, it is not cost-effective to collect goat milk everyday or every other day. In some areas, goat milk is collected only once each week, which is in violation of regulations specified in the Pasteurized Milk Ordinance for grade A milk. This study was conducted to determine the effect of up to 7 days of refrigerated bulk tank storage on composition, somatic cell count (SCC), pH, and microbiological quality of goat milk. Duplicate farm bulk tank samples were taken daily after the morning milking for seven consecutive days each month during the lactation season. Samples were analyzed immediately for all variables except free fatty acids. There were no significant changes (P > 0.05) detected in milk fat, protein, lactose, nonfat solids, SCC, or pH during extended storage, although significant effects of stage of lactation (P < 0.05) were observed. The mean standard plate count (SPC) increased to 1.8 x 10(5) CFU/ml after 6 days of storage, exceeding the grade A limit (i.e., 1.0 x 10(5) CFU/ml). The mean psychrotrophic bacteria count increased steadily to 1.5 x 10(4) CFU/ml after 6 days of storage, whereas the mean coliform count was approximately 500 CFU/ml for the first 3 days and less than 2500 CFU/ml throughout the 7 days of storage. No significant changes (P > 0.05) in the concentrations of free fatty acids, except for butyric and caprylic acids, were observed during milk storage. When stored under refrigerated and sanitary conditions, goat milk in farm bulk tanks met the grade A criteria for both SPC and SCC during 5 days of storage but was of low quality thereafter because of the growth of psychrotrophic bacteria.     

Effects of pasteurisation and high-pressure processing on vitamin C,tocopherols and fatty acids in mature human milk

Holder pasteurisation has traditionally been used in human milk banks to inactivate the pathogenic microorganisms and part of the commensal flora that are potentially present in milk from donors. However, this process results in variable loss of nutrients. In the search for an alternative to pasteurisation, we assessed the ability of high-pressure processing (HPP) to maintain the fatty acid, vitamin C and vitamin E contents of human milk and compared this process with Holder pasteurisation. Fatty acid proportions in milk, as well as levels of delta-, gamma-, and alpha-tocopherols did not vary with any of the treatments. Total vitamin C and ascorbic acid levels were maintained after HPP. However, after pasteurisation, total vitamin C and ascorbic acid were about 20% and 16% lower, respectively, than in untreated samples. These results suggest that HPP merits further investigation as a potential alternative to Holder pasteurisation for treating donor human milk.     

Microencapsulation of Lactobacillus paracasei by spray freeze drying

This study evaluates the implementation of a new process: spray freeze drying (SFD), to produce dry micro-capsules of Lactobacillus paracasei with high viability. The study concentrated on determining the survival of the cells, encapsulated in a matrix of maltodextrin and trehalose. SFD was compared with the conventional bulk freeze drying (BFD). Overall it was shown that SFD is a successful method to generate dry micro-capsules of probiotic cells with high viability (>60%). The spraying stage did not affect the viability of the bacteria. In the freezing stage, high osmotic pressures originated by elevated trehalose concentrations, helped preserving the cells viability. It was also found that the lower the maltodextrin molecular weight, the larger the beads volume and solids concentration, the higher is the bacteria survival during the freezing and drying stages. In the drying stage, trehalose concentration was also the critical factor that increased final probiotic viability.     

Physicochemical and functional properties of goat milk whey protein and casein obtained during different lactation stages

During lactation, goat milk contains colostrum, transitional milk, mature milk, and end milk. The protein present in goat milk during different lactation periods has different characteristics. This study aimed to characterize the protein profile of goat milk samples obtained at different lactation stages and to identify changes in the physicochemical and functional properties of whey protein and casein from goat milk collected at 1, 3, 15, 100, and 200 d after calving. The results demonstrated that the lactation period had a great influence on the physicochemical and functional properties of goat milk whey protein and casein, especially the protein properties of colostrum on the first day after delivery. The denaturation temperature, hydrophobicity, and turbidity of whey protein were significantly higher on the first day postpartum than at other lactation periods. Correspondingly, the colostrum whey protein also had better functional properties, such as emulsification, oil holding capacity, and foaming properties on the first day postpartum than at other lactation periods. For casein, the turbidity, particle size, water holding capacity, and foaming properties on the first day after delivery were significantly higher than those at other lactation periods, whereas the denaturation temperature, oil holding capacity, and emulsification followed the opposite trend. For both whey protein and casein, the 2 indicators of emulsifying properties, namely, emulsifying activity index and the emulsion stability, also followed an opposite trend relative to lactation stage, whereas the changes in foaming capacity with the lactation period were completely consistent with the change of foaming stability. These findings could provide useful information for the use of goat milk whey protein and casein obtained during different lactation stages in the dairy industry.     

Tracking Amazonian cheese microbial diversity: Development of an original,sustainable, and robust starter by freeze drying/spray drying

Marajó cheese made with raw buffalo milk in the Amazon region of Brazil can be considered a good source of wild lactic acid bacteria strains with unexplored and promising characteristics. The aim of this study was to develop a potential probiotic starter culture for industrial applications using freeze drying and spray drying. A decrease in the survival rates of freeze-dried samples compared with spray-dried samples was noted. The spray-dried cultures remained approximately 10⁹ cfu·g⁻¹, whereas the freeze-dried samples showed 10⁷ cfu·g⁻¹ after 60 d of storage at 4°C. All of the spray-dried samples showed a greater ability to decrease the pH in 10% skim milk over 24 h compared with the freeze-dried samples. The spray-dried samples showed a greater resistance to acidic conditions and to the presence of bile salts. In addition, under heat stress conditions, reduction was under 2 log cycles in all samples. Although the survival rate was similar among the evaluated samples after drying, the technological performance for skim milk showed some differences. This study may direct further investigations into how to preserve lactic acid bacteria probiotics to produce spray-dried starters that have a high number of viable cells that can then be used for industrial applications in a cost-effective way.