Influence of protein concentration and coagulation temperature on rennet-induced gelation characteristics and curd microstructure

This study characterized the coagulation properties and defined the cutting window (CW; time between storage modulus values of 35 and 70 Pa) using rheometry for milk standardized to 4, 5, or 6% protein and set at 28, 32, or 36°C. Milks were standardized to a protein-to-fat ratio of approximately 1 by blending ultrafiltration retentate, skim milk, and whole milk. The internal curd microstructure for selected curd samples was analyzed with transmission electron microscopy and scanning electron microscopy. Lowering the coagulation temperature caused longer rennet coagulation time and time to reach storage modulus of 35 Pa, translating into a wider CW. It also led to a lower maximum curd-firming rate (MCFR) with lower firmness at 40 min at a given protein level. Increasing protein levels resulted in the opposite effect, although without an effect on rennet coagulation time at a given temperature. On coagulation at 28°C, milk with 5% protein resulted in a similar MCFR (～4 Pa/min) and CW (～8.25 min) compared with milk with 4% protein at 32°C, which reflects more standard conditions, whereas increasing milk to 6% protein resulted in more than doubling of the curd-firming rate (MCFR = 9.20 Pa/min) and a shorter CW (4.60 min). Gels set at 28°C had lower levels of rearrangement of protein network after 40 min compared with those set at 36°C. Protein levels, on the other hand, had no influence on the levels of protein network rearrangement, as indicated by loss tangent values. The internal structure of curd particles, as investigated by both scanning electron microscopy and transmission electron microscopy, appeared to have less cross-linking and smaller casein aggregates when coagulated at 28°C compared with 36°C, whereas varying protein levels did not show a marked effect on aggregate formation. Overall, this study showed a marked interactive effect between coagulation temperature and protein standardization of milk on coagulation properties, which subsequently requires adjustment of the CW during cheesemaking. Lowering of the coagulation temperature greatly altered the curd microstructure, with a tendency for less syneresis during cutting. Further research is required to quantify the changes in syneresis and in fat and protein losses to whey due to changes in the microstructure of curd particles arising from the different coagulation conditions applied to the protein-fortified milk.     

Effects of technological settings on yield,curd, whey,and cheese composition during the cheese-making process from raw sheep milk in small rural dairies: Emphasis on cutting and cooking conditions

The technological conditions of cheese-making affect cheese yield and compound losses in the whey, especially the processes of cutting and cooking. Although significant compositional and functional differences have been reported among animal species, there is a lack of studies on the effects of cheese-making technology on cheese yield and losses for sheep milk. Thus, we examined the cheese-making settings in 8 small rural dairies working with raw sheep milk and their effects on cheese yield and compound losses in whey during the cheese-production season. Actual cheese yield varied in 2 to 3 kg of cheese/100 kg of milk among dairies due to the cheese-making conditions, particularly the duration of cutting and cooking and the final cooking temperature. The combination of the conditions used during cutting and cooking, especially, determined fat losses in the whey. Fat losses were increased with high-speed and short cutting time settings together with high stirring speed and long duration of cooking. Additionally, cheese-makers should adapt the cutting and cooking conditions to the seasonal variations of milk composition, especially during early summer, when fat losses in the whey are higher. Our results suggest that it could be useful to use approximately 10 to 15 min of cutting time and moderate cooking speed and duration. The data reported in this study may assist the improvement of the cheese-making process in small rural dairies using sheep milk, where facilities are limited and the role of the cheese-maker is crucial.     

Hot topic: microstructure quantification by scanning electron microscopy and image analysis of goat cheese curd

Five microstructural parameters of goat cheese curd (number of pores, their area and perimeter, strand thickness, and porosity) were studied by scanning electron microscopy and image analysis. Image analysis was used to characterize and quantify differences in all parameters and to provide a procedure for the measurement of strand thickness. The micrographs provided visual evidence of differences in the protein matrix and were quantified by image analysis at 3 production times: 34 ± 1 min (cutting), 154 ± 6 min (before molding), and 293 ± 35 min (after pressing). The data showed that this procedure is an adequate tool for quantifying differences in the parameters analyzed in industrial samples despite their natural heterogeneity. The procedure was reproducible and repetitive for the first 2 production times because no significant intragroup differences were observed. Significant differences were found when comparing the values of the microstructure parameters analyzed at 34 ± 1 min and those corresponding to 154 ± 6 min and 293 ± 35 min, but no significant differences between samples analyzed at 154 ± 6 min and 293 ± 35 min were found. All microstructure parameters analyzed were related at a significance level of at least 95%. This procedure enables the characterization of the microstructure of industrial goat cheese curd.     

Gastric digestion of milk protein ingredients: Study using an in vitro dynamic model

The coagulation behavior and the kinetics of protein hydrolysis of skim milk powder, milk protein concentrate (MPC), calcium-depleted MPC, sodium caseinate, whey protein isolate (WPI), and heated (90°C, 20 min) WPI under gastric conditions were examined using an advanced dynamic digestion model (i.e., a human gastric simulator). During gastric digestion, these protein ingredients exhibited various pH profiles as a function of the digestion time. Skim milk powder and MPC, which contained casein micelles, formed cohesive, ball-like curds with a dense structure after 10 min of digestion; these curds did not disintegrate over 220 min of digestion. Partly calcium-depleted MPC and sodium caseinate, which lacked an intact casein micellar structure, formed curds at approximately 40 min, and a loose, fragmented curd structure was observed after 220 min of digestion. In contrast, no curds were formed in either WPI or heated WPI after 220 min of digestion. In addition, the hydrolysis rates and the compositions of the digesta released from the human gastric simulator were different for the various protein ingredients, as detected by sodium dodecyl sulfate-PAGE. Skim milk powder and MPC exhibited slower hydrolysis rates than calcium-depleted MPC and sodium caseinate. The most rapid hydrolysis occurred in the WPI (with and without heating). This was attributed to the formation of different structured curds under gastric conditions. The results offer novel insights about the coagulation kinetics of proteins from different milk protein ingredients, highlighting the critical role of the food matrix in affecting the course of protein digestion.     

牛乳的凝固特性及其影响因素研究进展

本文综述了近年来有关牛乳凝固特性的研究进展,重点针对影响牛乳凝固能力的遗传和非遗传因素的研究进展进行了分析论述,包括不同来源牛乳的凝固特性、乳蛋白组分和工艺条件等对牛乳凝固能力的影响。旨在厘清影响牛乳凝固能力的主要因素。     

Effect of cutting time, temperature, and calcium on curd moisture, whey fat losses, and curd yield by response surface methodology

Response surface methodology was used to study the effect of temperature, cutting time, and calcium chloride addition level on curd moisture content, whey fat losses, and curd yield. Coagulation and syneresis were continuously monitored using 2 optical sensors detecting light backscatter. The effect of the factors on the sensors’ response was also examined. Retention of fat during cheese making was found to be a function of cutting time and temperature, whereas curd yield was found to be a function of those 2 factors and the level of calcium chloride addition. The main effect of temperature on curd moisture was to increase the rate at which whey was expelled. Temperature and calcium chloride addition level were also found to affect the light backscatter profile during coagulation whereas the light backscatter profile during syneresis was a function of temperature and cutting time. The results of this study suggest that there is an optimum firmness at which the gel should be cut to achieve maximum retention of fat and an optimum curd moisture content to maximize product yield and quality. It was determined that to maximize curd yield and quality, it is necessary to maximize firmness while avoiding rapid coarsening of the gel network and microsyneresis. These results could contribute to the optimization of the cheese-making process.     

Modeling of coagulation,curd firming,and syneresis of goat milk from 6 breeds

Traditional milk coagulation properties are used to predict the suitability of milk for cheese-making. In bovine and ovine species, the introduction of the concept of curd firming over time, continuously recorded by a lactodynamograph during prolonged tests, provides additional information about milk coagulation, curd-firming, and syneresis processes. The aims of present study were (1) to test the adaptability of a 4-parameter curd-firming model in the assessment of goat milk (also comparing published data of other species); (2) to describe variability of coagulation, curd firming, and syneresis processes among individual goat milk samples; (3) to quantify the effects of farm and animal factors (breed, parity, and stage of lactation); and (4) to compare 6 goat breeds for their model parameters. Milk samples from 1,272 goats reared in 35 farms were collected. Goats were of 6 breeds: Saanen and Camosciata delle Alpi for the Alpine type; and Murciano-Granadina, Maltese, Sarda, and Sarda Primitiva for the Mediterranean type. During a lactodynamographic analysis (60 min), 240 measures of curd firmness (mm) were recorded for each milk sample. The modeling of curd firming allowed us to achieve the rennet coagulation time estimated on the basis of all the data points (min); the curd firming and the curd syneresis instant rate constants; the asymptotical potential value of curd firming; the actual maximum curd firmness; and the time at which the curd firming maximum level is attained. Modeling parameter data were analyzed using a linear mixed model. Comparison with other dairy species showed several differences: goat milk coagulated later than sheep but earlier than bovine, and curd firming and curd syneresis instant rate constants were greater in small ruminants. Modeling parameters of goat milk were mostly affected by the farm effect (37% of the total variance, on average) compared with the results found for bovine and ovine samples, and this was probably attributable to the marked differences among goat farming systems. Small differences were demonstrated between Alpine and Mediterranean breeds, but the time of maximum curd firmness was lower in Murciano-Granadina compared with Maltese, Sarda, and Sarda Primitiva. Sarda and Sarda Primitiva were very similar and exhibited the most favorable coagulation properties of milk. For almost all the model parameters, the direct effect of breed was increased after correction for milk yield and composition. In conclusion, this approach allowed us to fully depict the effects of the different factors on coagulation of goat milk, and clarified the different renneting pattern among goat breeds, and with other species. Results could be used for the valorization of goat dairy products, also when these are linked to particular local breeds, and to stimulate further studies about relationships between coagulation and cheese-making traits.     

Ripening-related changes in Serra da Estrela cheese: a stereological study

Despite its relevance to sensory features and to fundamental explanation of the changes observed throughout cheese ripening, microstructural studies of specialty cheeses have lagged far behind those of industrialized cheeses. Hence, the purpose of this study was to pinpoint microstructural differences in the gel network of traditional Serra da Estrela cheese throughout ripening, using 2-dimensional image analysis, and to unfold correlations of such microstructural indicators with classical bulk chemical and textural parameters. Hence, samples were taken throughout the ripening period, following a nested design; uniform thin sections were systematically observed via light microscopy (LM, 200 ×) and transmission electron microscopy (TEM, 4,400 ×), and computer-assisted quantitative analysis of digital images was comprehensively performed following standard stereological methodology. Fresh cheeses exhibited the highest porosity and ratio of surface area to volume. Significant negative correlations were found between microstructural parameters and proteolysis indicators. Light microscopy images suggested that rearrangements exist, up to 21 d, of the cheese matrix that leave porosity and pores unchanged, whereas TEM images indicated a significant decrease in number of pores within the same time frame, especially those above 1 × 10⁻² μm² in area. The larger pores, chiefly with cross-sectional areas above 40 μm², were less represented by the end of ripening—and likely explain the observed significant decrease of cheese porosity without a change in number of pores. Field viewing significantly affected the microstructural parameter values, whereas section viewing affected significantly only LM-based ones. Categorical principal component analysis between the 2 types of microstructural data sets was performed, and permitted discrimination of each stage of ripening. Multiple linear regression analysis indicated that the variables associated with the nitrogen fraction were well predicted by stereological-based parameters (R² ≥ 0.96). Therefore, our findings demonstrate the potential of image analysis to monitor microstructure throughout ripening, and that the microstructure revealed by LM reflects more clearly cheese aging than that revealed by TEM.     

基于休闲豆干的热量成分检测及检测模型校正

以市售休闲豆干为原料,分别采用凯式定氮法、快速水分测定法和索氏提取法测定其蛋白质、水分和脂肪含量,再使用热量成分检测仪分别测定其含量,运用数学分析法对其检测模型进行校正,得到相应的校正模型。所得校正后的热量成分检测仪模型分别为:y_p=1.15x,y_w=0.85x,y_f=0.65x,模型适用于蛋白质含量、水分含量和脂肪含量范围分别为22.80~27.30,36.12~51.59,7.42~14.80g/100g的休闲豆干的校正。经校正后蛋白质含量平均绝对误差减小了1.44,平均误差率减小了6.23%;水分含量平均绝对误差减小了6.77,平均误差率减小了14.71%;脂肪含量平均绝对误差减小了4.49,平均误差率减小了45.74%,校正后热量成分检测仪能够快速、准确地检测休闲豆干中蛋白质、水分、脂肪的含量,为休闲豆干蛋白质、水分、脂肪含量的快速检测提供了新途径。     

Kinetics of Formation and Physicochemical Characterization of Thermally-Induced β-Lactoglobulin Aggregates

Abstract: The kinetics of heat denaturation and aggregation for β-lactoglobulin dispersions (5% w/v) were studied at 3 pHs (6, 6.4, and 6.8) and at a heating temperature of 80 °C. Protein aggregates were characterized for hydrodynamic diameter, microstructure, and molecular weight by means of dynamic light scattering, transmission electron microscopy, and polyacrylamide gel electrophoresis, respectively. Concentration of native β-lactoglobulin decreased with holding time and with a decrease in the pH. Apparent rate constants were calculated for β-lactoglobulin denaturation applying the general kinetic equation solved for a reaction order of 1.5. Values of the apparent reaction rate constant k = 7.5, 6.3 and 5.6 × 10⁻³ s⁻¹ were found for pH 6, 6.4, and 6.8, respectively. Decreasing the pH of the dispersions produced higher aggregate sizes. After a holding time of 900 s, average hydrodynamic diameters for β-lactoglobulin aggregates at pH 6, 6.4, and 6.8 were 96, 49, and 42 nm, respectively. These results were confirmed by transmission electron microscopy images, where a shift in the size and morphology of aggregates was found, from large and spherical at pH 6 to smaller and linear aggregates at pH 6.8. β-Lactoglobulin formed disulfide-linked intermediates (dimers, trimers, tetramers) and so on) which then formed high molecular weight aggregates. From the results obtained by DLS, TEM, and SDS-PAGE a mechanism for β-lactoglobulin aggregation was proposed. This study shows that heat treatment can be used to produce protein aggregates with different sizes and morphologies to be utilized as ingredients in foods.     

Goat farm variability affects milk Fourier-transform infrared spectra used for predicting coagulation properties

Driven by the large amount of goat milk destined for cheese production, and to pioneer the goat cheese industry, the objective of this study was to assess the effect of farm in predicting goat milk-coagulation and curd-firmness traits via Fourier-transform infrared spectroscopy. Spectra from 452 Sarda goats belonging to 14 farms in central and southeast Sardinia (Italy) were collected. A Bayesian linear regression model was used, estimating all spectral wavelengths' effects simultaneously. Three traditional milk-coagulation properties [rennet coagulation time (min), time to curd firmness of 20 mm (min), and curd firmness 30 min after rennet addition (mm)] and 3 curd-firmness measures modeled over time [rennet coagulation time estimated according to curd firmness change over time (RCT_eq), instant curd-firming rate constant, and asymptotical curd firmness] were considered. A stratified cross validation (SCV) was assigned, evaluating each farm separately (validation set; VAL) and keeping the remaining farms to train (calibration set) the statistical model. Moreover, a SCV, where 20% of the goats randomly taken (10 replicates per farm) from the VAL farm entered the calibration set, was also considered (SCV₈₀). To assess model performance, coefficient of determination (R²_VAL) and the root mean squared error of validation were recorded. The R²_VAL varied between 0.14 and 0.45 (instant curd-firming rate constant and RCT_eq, respectively), albeit the standard deviation was approximating half of the mean for all the traits. Although average results of the 2 SCV procedures were similar, in SCV₈₀, the maximum R²_VAL increased at about 15% across traits, with the highest observed for time to curd firmness of 20 mm (20%) and the lowest for RCT_eq (6%). Further investigation evidenced important variability among farms, with R²_VAL for some of them being close to 0. Our work outlined the importance of considering the effect of farm when developing Fourier-transform infrared spectroscopy prediction equations for coagulation and curd-firmness traits in goats.     

Modeling rennet coagulation time and curd firmness of milk

Milk coagulation properties (MCP) are traditionally expressed using rennet coagulation time (RCT), time to curd firmness (CF) of 20 mm (k₂₀), and CF 30 min after enzyme addition (a₃₀) values, all of which are single-point measures taken from the output of computerized renneting meters, such as the Formagraph. Thus, traditional MCP use only some of the available information. Moreover, because of the worldwide spreading of breeds such as the Holstein-Friesian, characterized by late-coagulating milk, it happens often that some samples do not coagulate at all, that a₃₀ is strongly and negatively related to RCT, and that k₂₀ is not measurable. The aim of the present work was to model CF as a function of time (CF_t, mm) over a 30-min interval. The model tested was CFt=CFP×(1−e−kCF×(t−RCT)), where CF_P (mm) is the potential asymptotical CF at an infinite time, k_CF (min⁻¹) is the curd firming rate constant, and RCT is measured in minutes. The CF_t model was initially applied to data of milk of each of 105 Brown Swiss cows from 7 herds, each sampled once (trial 1). Four samples did not coagulate within 30 min. Eighty-seven of the 101 individual equations obtained fit the CF data of milk samples very well, even though the samples differed in composition, and were produced by cows of different ages and days in milk, reared on different farms (coefficient of determination >0.99; average residual standard deviation = 0.21 mm). Samples with a very late RCT (slowly coagulating samples) yielded so few observational data points that curve parameters could not be precisely estimated. The repeatability of CF_t equation parameters was estimated using data obtained from 5 replicates of each of 2 samples of bulk milk from 5 Holstein-Friesian cows analyzed every day for 5 consecutive days (trial 2). Repeatability of RCT was better than that of the other 2 parameters. Moreover, traditional MCP values (RCT, a₃₀, and k₂₀) can be obtained from the individual CF_t equations, using all available information. The MCP estimated from equations were very similar to the single-point measures yielded by the computerized renneting meter (coefficient of determination >0.97), but repeatability was slightly better. The model allowed the estimation of k₂₀ for samples with a very late coagulation or with very slow curd firming. Finally, the 3 novel parameters used to assess different milk samples were less interdependent than are the traditional measures, and their practical and scientific utility requires further study.     

3种乳源酪蛋白粒径及胶束结构的差异性

通过纳米粒度分析仪和扫描电子显微镜,分别对水牛乳、牛乳及羊乳中的酪蛋白颗粒直径大小分布情况及酪蛋白胶束结构进行研究。结果表明：水牛乳、牛乳及羊乳中酪蛋白的粒径分布及胶束结构方面存在明显的差异。水牛乳酪蛋白平均颗粒直径为182.3nm,酪蛋白颗粒互相连接成较细长的胶束,胶束之间交联成网络状;牛乳酪蛋白平均颗粒直径为207.4nm,酪蛋白颗粒聚集成直径较大的胶束;羊乳酪蛋白平均颗粒直径为173.8nm,酪蛋白颗粒仅能够形成较短的胶束,也不能交联成网络状。     

关于腐乳的感官品评方法

腐乳是中华民族独特的调味品，介绍了腐乳品评的基本要求，品评标准，打分标准。     

影响卷烟端部落丝量的因素研究

通过卷制结构不同的烟丝、测试通过卷接设备的烟丝结构、调整设备参数及喂丝方式等实验，发现烟丝的结构和耐加工性能是影响卷烟端部落丝量的主要因素，卷接设备的参数调整、喂丝方式是影响卷烟端部落丝量的次要因素。     

Breed of cow and herd productivity affect milk composition and modeling of coagulation,curd firming,and syneresis

Milk coagulation properties (MCP) have been widely investigated in the past using milk collected from different cattle breeds and herds. However, to our knowledge, no previous studies have assessed MCP in individual milk samples from several multi-breed herds characterized by either high or low milk productivity, thereby allowing the effects of herd and cow breed to be evaluated independently. Multi-breed herds (n = 41) were classified into 2 categories based on milk productivity (high vs. low), defined according to the average milk net energy yielded daily by lactating cows. Milk samples were taken from 1,508 cows of 6 different breeds: 3 specialized dairy (Holstein-Friesian, Brown Swiss, Jersey) and 3 dual-purpose (Simmental, Rendena, Alpine Grey) breeds, and analyzed in duplicate (3,016 tests) using 2 lactodynamographs to obtain 240 curd firming (CF) measurements over 60 min (1 every 15 s) for each duplicate. The 5 traditional single-point MCP (RCT, k₂₀, a₃₀, a₄₅, and a₆₀) were yielded directly by the instrument from the available CF measures. All 240 CF measures of each replicate were also used to estimate 4 individual equation parameters: RCT estimated according to curd firm change over time modeling (RCT_eq), asymptotic potential curd firmness (CF_P), curd firming instant rate constant (k_CF), and syneresis instant rate constant (k_SR) and 2 derived traits: maximum curd firmness achieved within 45 min (CF_max) and time at achievement of CF_max (t_max) by curvilinear regression using a nonlinear procedure. Results showed that the effect of herd-date on traditional and modeled MCP was modest, ranging from 6.1% of total variance for k₂₀ to 10.7% for RCT, whereas individual animal variance was the highest, ranging from 32.0% for t_max to 82.5% for RCT_eq. The repeatability of MCP was high (>80%) for all traits except those associated with the last part of the lactodynamographic curve (i.e., a₆₀, k_SR, k_CF, and t_max: 57 to 71%). Reproducibility, taking into account the effect of instrument, was equal to or slightly lower than repeatability. Milk samples collected in farms characterized by high productivity exhibited delayed coagulation (RCT_eq: 18.6 vs. 16.3 min) but greater potential curd firmness (CF_P: 76.8 vs. 71.9 mm) compared with milk samples collected from low-productivity herds. Parity and days in milk influenced almost all MCP. Large differences in all MCP traits were observed among breeds, both between specialized and dual-purpose breeds and within these 2 groups of breeds, even after adjusting for milk quality and yield. Milk quality and MCP of samples from Jersey cows, and coagulation time of samples from Rendena cows were better than in milk from Holstein-Friesian cows, and intermediate results were found with the other breeds of Alpine origin. The results of this study, taking into account the intrinsic limitation of this technique, show that the effects of breed on traditional and modeled MCP are much greater than the effects of herd productivity class, parity, and DIM. Moreover, the variance in individual animals is much greater than the variance in individual herds within herd productivity class. It seems that improvement in MCP depends more on genetics (e.g., breed, selection) than on environmental and management factors.