Relationship Between Maternal Age and Macronutrient Content of Colostrum.

BACKGROUND: Little is known about the relationship between maternal age and the macronutrient content of colostrum. RESEARCH AIMS: This study aimed to evaluate the relationship between maternal age and human milk macronutrient content by comparing the concentrations of lactose, proteins, and lipids in the colostrum of women with younger, moderate, and advanced maternal age. METHODS: An observational, cross-sectional study was designed to compare the macronutrient concentrations in the colostrum of women aged < 20 years, 20 to 34 years, and > 34 years (younger, moderate, and advanced maternal age, respectively; n = 33 per group). For each participant, 3 ml of colostrum was collected by manual extraction from the right breast at 10 am, 39-48 hr after delivery, and analyzed using a Miris Human Milk Analyzer. Macronutrient concentrations were compared between the groups using analysis of variance. P < 0.05 was considered significant. RESULTS: Mothers with moderate maternal age had a higher colostrum lipid concentration than those with younger or advanced maternal age (2.3 mg, SD = 1.4 mg vs. 1.5 mg, SD = 1.0 mg vs. 1.6 mg, SD = 0.9 mg, respectively; p = 0.007). Lactose and protein contents in the analyzed samples did not differ among the three study groups. CONCLUSION: This study lends support to the potential variation of lipids in colostrum by maternal age and suggests individual adaptation to the nutritional components of milk to the needs of the infant may be beneficial.

Genome-scale metabolic modeling of the human milk oligosaccharide utilization by Bifidobacterium longum subsp. infantis.

Bifidobacterium longum subsp. infantis is a representative and dominant species in the infant gut and is considered a beneficial microbe. This organism displays multiple adaptations to thrive in the infant gut, regarded as a model for human milk oligosaccharides (HMOs) utilization. These carbohydrates are abundant in breast milk and include different molecules based on lactose. They contain fucose, sialic acid, and N-acetylglucosamine. Bifidobacterium metabolism is complex, and a systems view of relevant metabolic pathways and exchange metabolites during HMO consumption is missing. To address this limitation, a refined genome-scale network reconstruction of this bacterium is presented using a previous reconstruction of B. infantis ATCC 15967 as a template. The latter was expanded based on an extensive revision of genome annotations, current literature, and transcriptomic data integration. The metabolic reconstruction (iLR578) accounted for 578 genes, 1,047 reactions, and 924 metabolites. Starting from this reconstruction, we built context-specific genome-scale metabolic models using RNA-seq data from cultures growing in lactose and three HMOs. The models revealed notable differences in HMO metabolism depending on the functional characteristics of the substrates. Particularly, fucosyl-lactose showed a divergent metabolism due to a fucose moiety. High yields of lactate and acetate were predicted under growth rate maximization in all conditions, whereas formate, ethanol, and 1,2-propanediol were substantially lower. Similar results were also obtained under near-optimal growth on each substrate when varying the empirically observed acetate-to-lactate production ratio. Model predictions displayed reasonable agreement between central carbon metabolism fluxes and expression data across all conditions. Flux coupling analysis revealed additional connections between succinate exchange and arginine and sulfate metabolism and a strong coupling between central carbon reactions and adenine metabolism. More importantly, specific networks of coupled reactions under each carbon source were derived and analyzed. Overall, the presented network reconstruction constitutes a valuable platform for probing the metabolism of this prominent infant gut bifidobacteria.IMPORTANCEThis work presents a detailed reconstruction of the metabolism of Bifidobacterium longum subsp. infantis, a prominent member of the infant gut microbiome, providing a systems view of its metabolism of human milk oligosaccharides.

Evaluation of a novel stationary phase for the separation of lactose and isomers in lactose-free products using high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD).

Lactose intolerance is a widespread condition, which prevents a large number of people from consuming dairy products as a part of their daily diet. It is estimated that an average of 65% of the global population is suffering from lactose intolerance. The global market for 'lactose-free' dairy products is rapidly growing and the criteria for 'lactose-free' labelled products are becoming stricter. To check the lactose contents in these products there is a need for fast, sensitive, and selective analytical method. A method is presented for fast and sensitive determination of lactose and its isomers using High-Performance Anion Exchange Chromatography in combination with Pulsed Amperometric Detection (HPAEC-PAD). The use of a new anion-exchange column, SweetSep™ AEX200, which is a strong anion-exchange column with highly monodisperse 5 µm particles, allowed the separation of all compounds of interest in less than 8 min with high resolution. A variety of dairy products were analyzed to demonstrate the versatility of the method.

Characterization of lyophilized equine colostrum.

Foals require maternal colostrum in the first hours of life to prevent failure of transfer of passive immunity (FTIP). Innovative storage methods such as lyophilization may enable conservation of colostrum immunoglobulins by a differentiated process of dehydration. The current study aimed to compare the quality of equine colostrum after freezing and after the lyophilization process. Thirty-one pregnant Quarter Horse mares were used. The IgG concentration of frozen and lyophilized colostrum was determined by simple radial immunodiffusion (SRID) and Brix refractometry. The physical-chemical composition (pH, total protein (TP), fat, lactose, salts, total solids (TS), and density) of the samples was evaluated and the lyophilized colostrum reconstitution test was performed. There were no significant differences (P > 0.05) in the variables IgG, fat, lactose, salts, TS, density, and pH between samples measured before and after lyophilization. There was a significant difference (P < 0.05) between the Brix average and the TP of the frozen and lyophilized colostrum samples. Lyophilization resulted in a small reduction (6.55%) in the IgG concentration measured by SRID. A strong positive correlation was observed between colostrum density and IgG concentration by SRID (r = 0.76) and between Brix and IgG concentration by SRID (r = 0.77). In the reconstitution test, the lyophilized colostrum was easily rehydrated in water, with full dilution, and remained stable. Lyophilization could be an alternative for the conservation of mare colostrum, since it is a very efficient process for retaining the physicochemical characteristics of the product, with minimal loss, particularly of IgG.

Milk beverage base with lactose removed with ultrafiltration: Effect of fat and protein concentration on sensory and physical properties.

Our objectives were to determine the effect of fat (skim to whole milk) and protein (3.4%-10.5%) concentration on the sensory and physical properties of milk beverage base that had lactose and other low molecular components removed by ultrafiltration (UF). In experiment 1, a matrix of 16 treatments was produced to achieve 4 levels of lactose removal (0%, 30%, 70%, and 97%) at each of 4 fat levels (skim, 1%, 2%, and whole milk). In experiment 2, a matrix of 12 treatments was produced to achieve 4 levels of lactose removal (0%, 30%, 70%, and 97%) at each of 3 protein concentrations (3.4%, 6.5%, and 10.5% protein). Physical and sensory properties of these products were determined. Removal of >95% of milk lactose by UF required a diafiltration volume of approximately 3 times the milk volume. Lactose and low molecular weight solute removal increased whiteness across the range from skim to whole milk while decreasing viscosity and making milk flavor blander. In addition, lactose (and other low molecular weight solute) removal by UF decreased titratable acidity by more than 50% and increased milk pH at 20°C to >7.0. Future work on milk and milk-based beverages with lactose removed by UF needs to focus on interaction of the remaining milk solids with added flavorings, changing casein to whey protein ratio before removal of lactose by UF, and the effect of lactose and low molecular weight solute removal on heat stability, particularly for neutral-pH, shelf-stable milk-based beverages.

Porungo cheese whey: a new substrate to produce ß-galactosidase.

The bioconversion of porungo cheese whey to produce ß-galactosidase in batch system was studied. The whey released after curd cutting and precipitation during porungo cheese production was collected in borosilicate flasks. Two strains of Kluyveromyces marxianus, CCT 4086 and CBS 6556, and whey supplementation with different nitrogen sources were evaluated. Different temperatures (30 °C and 37 °C) and pH values (5.0 to 7.0) were investigated to establish the best conditions for enzyme production. The highest enzymatic activity was obtained by K. marxianus CCT 4086 in porungo cheese whey supplemented with yeast extract (16.73 U mL-1). K. marxianus CCT 4086 produced superior ß-galactosidase activity when compared to CBS 6556 for all media tested (ranging from 11.69 to 14.40 U mL-1). Highest ß-galactosidase activity was reached under conditions of pH 7.0 and 30 °C using K. marxianus CCT 4086 in the better media composition. The lowest enzymatic activity was observed at 37 °C for all pH values tested (10.69 U mL-1 to 13.94 U mL-1) and a highest ß-galactosidase activity was reached in pH 7.0 for both two temperatures (11.42 to 15.93 U mL-1). Porungo cheese whey shows potential for industrial ß-galactosidase production by microbial fermentation.

Association between hyperketolactia and production in early-lactating dairy cows.

Study aims were to investigate associations of hyperketolactia (HYKL) status of Holstein dairy cows between 6 and 60 d in milk (DIM), defined by milk acetone (mACE) and ß-hydroxybutyrate (mBHB) content, with daily milk yield and composition. Milk samples (∼5.0 million) were collected over a 5-yr period (2014-2019) within the milk recording system in Poland. Concentrations of mACE and mBHB determined by Fourier-transform infrared spectroscopy were used to categorize samples into 4 ketolactia groups. Based on threshold values of ≥0.15 mmol/L mACE and ≥0.10 mmol/L mBHB, ketolactia groups were normoketolactia (NKL; mACE <0.15 mmol/L and mBHB <0.10 mmol/L), BHB hyperketolactia (HYKLBHB; mACE <0.15 mmol/L and mBHB ≥0.10 mmol/L), ACE hyperketolactia (HYKLACE; mACE ≥0.15 mmol/L and mBHB <0.10 mmol/L), and ACE and BHB hyperketolactia (HYKLACEBHB; mACE ≥0.15 mmol/L and mBHB ≥0.10 mmol/L). To investigate ketolactia association with production outcomes, a linear model was developed, including ketolactia group, DIM, parity, their interactions, year-season as fixed effects, and random effects of herd and cow. Among all milk samples, 31.2% were classified as HYKL, and of these, 52.6%, 39.6%, and 7.8% were HYKLACEBHB, HYKLBHB, and HYKLACE, respectively. Ketolactia groups differed for all traits studied in all parities and DIM. Among HYKL groups, lowest milk yield was found in HYKLACEBHB cows, except for 6 to 30 DIM in first- and second-lactation cows. Milk yield of HYKLBHB cows was higher than that of NKL cows until 20 to 30 DIM, and then it was lower than NKL cows. Milk yield of HYKLACE cows was mostly lower than NKL cows. Energy-corrected milk (ECM) yield of HYKLACEBHB cows was higher than that of NKL cows until 30 to 35 DIM for second lactation and third lactation or greater, and in the whole study period for first lactation. The yield of ECM for HYKLBHB cows was mostly higher than that of NKL cows, whereas HYKLACE cows had higher ECM than NKL cows until 15 to 25 DIM and then was lower for the HYKLACE group. Milk composition differed among HYKL groups. Highest milk fat (MF) and lowest milk lactose (ML) contents were observed in HYKLACEBHB cows. Cows in HYKLACEBHB and HYKLBHB groups had higher MF and lower milk protein (MP; except in 6-8 DIM in first lactation) and ML content than NKL cows. Milk fat content was higher in HYKLACE than NKL cows in first lactation and during the first 30 to 40 DIM in older cows. Lactose content was lower in HYKLACE than in NKL cows within 30 to 40 DIM; afterward it was higher in NKL cows. Lower MP content was found in HYKLACE than in NKL cows, except during 6 to 9 DIM for cows in first lactation and third lactation or greater. In conclusion, HYKL is associated with altered milk production in all parities, but a range of these negative relations depends on ketone status addressing both ACE and BHB contents. Further research is needed to ascertain underpinning biochemical defects of HYKL from elevated ACE, alone or in combination with BHB, during early lactation.

Imputation of missing milk Fourier transform mid-infrared spectra using existing milk spectral databases: A strategy to improve the reliability of breeding values and predictive models.

The use of milk Fourier transform mid-infrared (FT-MIR) spectrometry to develop management and breeding tools for dairy farmers and industry is growing and supported by the availability of numerous new predicted phenotypes to assess the nutritional quality of milk and its technological properties, but also the animal health and welfare status and its environmental fingerprint. For genetic evaluations, having a long-term and representative spectral dairy herd improvement (DHI) database improves the reliabilities of estimated breeding values (EBV) from these phenotypes. Unfortunately, most of the time, the raw spectral data used to generate these estimations are not stored. Moreover, many reference measurements of those phenotypes, needed during the FT-MIR calibration step, are available from past research activities but lack spectra records. So, it is impossible to use them to improve the FT-MIR models. Consequently, there is a strong interest in imputing those missing spectra. The innovative objective of this study was to use the existing large spectral DHI database to estimate missing spectra by selecting probable spectra using, as the match criteria, common dairy traits recorded for a long time by DHI organizations. We tested 4 match criteria combinations. Combination 1 required to have equal fat and protein contents between the sample for which a spectrum was to be estimated and the reference samples in the DHI database. Combination 2 also required an equal urea content. Combination 3 requested equal fat, protein, and lactose contents. Finally, combination 4 included all criteria. When more than one spectrum was found during the search, their average was the estimated spectrum for the query sample. Concretely, this study estimated missing spectra for 1,700 samples using 2,000,000 spectral DHI records. For assessing the effect of this spectral estimation on the prediction quality, FT-MIR equations were used to predict 11 phenotypes, selected as their quantification used different FT-MIR regions. They were related to the milk fat and mineral composition, lactoferrin content, quantity of eructed methane, body weight (BW), and dry matter intake. The accuracy between predictions obtained from actual and estimated spectra was evaluated by calculating the mean absolute error (MAE). The criteria in the fourth and second combinations were too strict to estimate a spectrum for most samples. Indeed, for many samples, no spectra with the same values for those matching criteria was found. The third match criteria combination had a poorer prediction performance for all studied traits and spectral absorptions than the first combination due to fewer matched samples available to compute the missing spectrum. By allowing a range for matching lactose content (±0.1 g/dL milk), we showed that this new combination increased the number of selected samples to compute missing spectra and predict better the infrared absorption at different wavenumbers, especially those related to the lactose quantification. The prediction performance was further improved by performing queries on the entire Walloon DHI spectral database (6,625,570 spectra), and it varied among the studied phenotypes. Without considering the traits used for the matching, the best predictions were obtained for the content of saturated fatty acids (MAE = 0.15 g/dL milk) and BW (MAE = 12.80 kg). Yet, the predictions for the unsaturated fatty acids were less accurate (MAE = 0.13 and 0.018 g/dL milk for monounsaturated and polyunsaturated fatty acids), likely because of the poorer predictions of spectral regions related to long-chain fatty acids. Similarly, poorer predictions were observed for the amount of methane eructed by dairy cows (MAE = 47.02 g/d), likely because it is not directly related to fat content or composition. Prediction accuracies for the remaining traits were also low. In conclusion, we observed that increasing the number of relevant matching criteria helps improve the quality of FT-MIR predicted phenotypes and the number of spectra used during the search. So, it would be of great interest to test in the future the suitability of the developed methodology with large-scale international spectral databases to improve the reliability of EBV from these FT-MIR-based phenotypes and the robustness of FT-MIR predictive models.

Measuring ß-galactosidase activity in opaque dairy solutions under optimum conditions for galactooligosaccharide synthesis by isothermal titration calorimetry.

The dairy industry uses enzymes to make cheese, alter product flavor, and eliminate lactose. The activities of these enzymes have been measured in clear buffered solutions, but because of the limitations of spectrophotometric methods, enzyme activities have not been measured in opaque or colored dairy products where they are used. Isothermal titration calorimetry (ITC) can be used to determine reaction kinetics in opaque and colored solutions by measuring the heat rate (thermal power) from enzyme-catalyzed reactions as a function of time. This study used ITC to measure ß-galactosidase activity in opaque solutions of milk, sweet whey, sweet whey permeate, acid whey, and acid whey permeate with 2 ß-galactosidase (Enzyme Commission number 3.2.1.23) isozymes derived from Aspergillus oryzae and Kluyveromyces lactis. The components of the dairy fluids alter the enzyme kinetics and reaction thermodynamics, and the reactions catalyzed by the 2 homologues differ as shown by differing thermodynamic profiles. The study demonstrates that ITC can be used to measure enzyme activity in opaque and colored dairy fluids and identify reactions by their thermodynamic properties.

Detection of two SNPs of the LIPE gene in Holstein-Friesian cows with divergent milk production.

The LIPE gene (lipase E, hormone-sensitive type), also known as hormone-sensitive lipase, acts as a primary regulator of lipid metabolism during lactation in cows. We studied a total of two hundred Holstein-Friesian cows and performed sequencing analysis that revealed two synonymous nucleotide changes within the LIPE gene: a transition change, c.276 T > C in exon 2 (g.50631651 T > C; position 351 of GenBank: ON638900) and a transversion change, c.219C > A in exon 6 (g.50635369C > A; position 1070 of GenBank: ON638901). The observed genotypes were TC and CC for the c.276 T > C SNP and CC and CA for the c.219C > A SNP. Notably, the heterozygous TC genotype of the T351C SNP exhibited a significant association with high milk yield. Furthermore, the T351C SNP displayed significant associations with various milk parameters, including temperature, freezing point, density and the percentages of fat, protein, lactose, solids and solids-not-fat, with the homozygous CC genotype showing higher values. The c.219C > A SNP also demonstrated a significant association with milk composition, with heterozygous genotypes (CA) exhibiting higher percentages of fat, protein, and lactose compared to homozygous genotypes (CC). This effect was consistent among both high and low milk producers for fat and lactose percentages, while high milk producers exhibited a higher protein percentage than low milk producers. These findings highlight the importance of considering the detected SNPs in marker-assisted selection and breeding programs for the identification of high milk-producing Holstein-Friesian cows and potentially other breeds. Moreover, this study strongly supports the fundamental role of the LIPE gene in milk production and composition in lactating animals.

Recovery of lactose from simulated delactosed whey permeate by a low-temperature crystallization process.

Delactosed whey permeate is the mother liquor/by-product of lactose manufacture, but it still contains around 20 wt% lactose. The high mineral content, stickiness, and hygroscopic behavior prevent further recovery of lactose in the manufacturing process. Therefore, its use is currently limited to low-value applications such as cattle feed, and more often it is seen as waste. This study investigates a new separation technique operating at sub-zero conditions. At low temperature, precipitation of calcium phosphate is expected to be reduced and the lower solubility at sub-zero temperature makes it possible to recover a large portion of the lactose. We found that lactose could be crystallized at sub-zero conditions. The crystals had a tomahawk morphology and an average size of 23 and 31 µm. In the first 24 h, the amount of calcium phosphate precipitated was limited, whereas the lactose concentration was already close to saturation. The overall rate of crystallization was increased compared with the crystals recovered from a pure lactose solution. Mutarotation was rate limiting in the pure system but it did not limit the crystallization of lactose from delactosed whey permeate. This resulted in faster crystallization; after 24 h the yield was 85%.

Unveiling the attributes of rabbit milk.

Increasing the knowledge of rabbit milk can help in breeding practice to solve issues considering the health and growth of rabbit kits. The goal of the study was to perform a broad physicochemical analysis of rabbit milk and examine the effect of the reproductive status of the females on daily milk yield and milk attributes. The study was conducted on a commercial rabbit farm and included three consecutive lactations of Hycole does. It has been observed that the daily milk production increased from the 2nd till the 14th day of lactation when does produced almost 300 g of milk daily. The day of lactation caused a significant variation in the content of total solids, solids-not-fat, total protein, casein, lactose, C18: 2, C18: 3, Somatic Cell Count, and pH. The percentage of fat globules categorised according to their diameter changed with the ongoing lactation as well, and the diameter increased from 5 to 7 µm. The percentage of small milk fat globules decreased with lactation day, causing a possible decrease in the digestions rates of milk. Pregnancy had a negative impact on milk production, kits growth performance, and the content of total protein, solids-not-fat, and lactose in milk. Therefore, we can speculate about the negative impact of overlapping lactations and pregnancies on rabbit kits, as their growth is dependent on milk production and composition.

Calcium dynamics and associated temporal patterns of milk constituents in early-lactation multiparous Holsteins.

At the onset of lactation, calcium (Ca) homeostasis is challenged. For the transitioning dairy cow, inadequate responses to this challenge may result in subclinical hypocalcemia at some point in the postpartum period. It has been proposed that dynamics of blood Ca and the timing of subclinical hypocalcemia allow cows to be classified into 4 Ca dynamic groups by assessing serum total Ca concentrations (tCa) at 1 and 4 days in milk (DIM). These differing dynamics are associated with different risks of adverse health events and suboptimal production. Our prospective cohort study aimed to characterize the temporal patterns of milk constituents in cows with differing Ca dynamics to investigate the potential of Fourier-transform infrared spectroscopic (FTIR) analysis of milk as a diagnostic tool for identifying cows with unfavorable Ca dynamics. We sampled the blood of 343 multiparous Holsteins on a single dairy in Cayuga County, New York, at 1 and 4 DIM and classified these cows into Ca dynamic groups using threshold concentrations of tCa (1 DIM: tCa <1.98 mmol/L; 4 DIM: tCa <2.22 mmol/L) derived from receiver operating characteristic curve analysis based on epidemiologically relevant health and production outcomes. We also collected proportional milk samples from each of these cows from 3 to 10 DIM for FTIR analysis of milk constituents. Through this analysis we estimated the milk constituent levels of anhydrous lactose (g/100 g of milk and g/milking), true protein (g/100 g of milk and g/milking), fat (g/100 g of milk and g/milking), milk urea nitrogen (mg/100 g of milk), fatty acid (FA) groups including de novo, mixed origin, and preformed FA measured in grams/100 g of milk, by relative percentage, and grams/milking, as well as energy-related metabolites including ketone bodies and milk-predicted blood nonesterified FA. Individual milk constituents were compared among groups at each time point and over the entire sample period using linear regression models. Overall, we found differences among the constituent profiles of Ca dynamic groups at approximately every time point and over the entire sample period. The 2 at-risk groups of cows did not differ from each other at more than one time point for any constituent, however prominent differences existed between the milk of normocalcemic cows and the milk of the other Ca dynamic groups with respect to FA. Over the entire sample period, lactose and protein yield (g/milking) were lower in the milk of at-risk cows than in the milk of the other Ca dynamic groups. In addition, milk yield per milking followed patterns consistent with previous Ca dynamic group research. Though our use of a single farm does limit the general applicability of these findings, our conclusions provide evidence that FTIR may be a useful method for discriminating between cows with different Ca dynamics at time points that may be relevant in the optimization of management or development of clinical intervention strategies.

Hydration dominates dielectric spectroscopy-based high accuracy quantification of lactose content in cow's milk.

BACKGROUND: Lactose is a critical factor in the quality of milk and dairy products. Achieving high accuracy and rapid detection of lactose content in cow's milk remains a challenge. Dielectric spectroscopy has emerged as a promising tool for detecting food components. We explored the effect of lactose content on the dielectric spectra of cow's milk and we propose a rapid analytical method for the quantitative determination of lactose content in cow's milk with high accuracy based on dielectric spectra. RESULTS: We obtained the dielectric spectra of 316 cow's milk samples in the frequency range 20-4500 MHz and noticed a strong negative correlation between the lactose content and the value of the dielectric loss factor (ε″) below 1500 MHz. Lactose does not affect cow's milk dielectric properties by excluded volume effect, but dominates the effect on the dielectric properties of cow's milk by hydration. The support vector regression model based on the variable importance in projection has the best prediction performance for lactose content. Its root-mean-square error of prediction set and residual prediction deviation is 0.29 g kg-1 and 6.968, respectively, and its prediction performance is better than that of the currently reported near-infrared (NIR) method and other methods. CONCLUSION: Despite the weak polarity of lactose molecules, its hydration is a significant factor affecting the dielectric properties of milk. The present study provides a basis for high accuracy and rapid quantitative detection of lactose in cow's milk based on dielectric spectra. © 2023 Society of Chemical Industry.

Lactose and Galactose Content in Spanish Cheeses: Usefulness in the Dietary Treatment of Patients with Galactosaemia.

In galactosaemia, a strict galactose-free diet is necessary to prevent or resolve acute symptoms in infants. However, because the body produces up to 10 times more galactose than is found in a galactose-restricted diet, excessively restrictive diets should be avoided in children and adults to prevent nutritional deficiencies. Since cheese is a nutritional source of the calcium necessary for bone health, the latest international guidelines on the management of classical galactosaemia (2017) allow the consumption of cured cheeses with less than 25 mg of galactose/100 g and recommend that each country verifies the adequacy of the cheeses, since most mature cheeses do not always have a lower galactose content. In total, 32 cheese samples were purchased (19 Spanish and 13 international cheeses), and their lactose and galactose contents were analysed using ion chromatography with pulsed amperometric detection (IC-PAD). Five Spanish cheeses contained less than 25 mg of galactose/100 g: García Baquero semi-cured cheese; Hacendado, Gran Reserva and Mahón cured cheeses; and García Baquero Reserva 12-month cured cheese. In addition, eight international cheeses were confirmed as suitable: Comté, Gouda, Gruyere, Maasdam, Parmigiano, Edam, Emmental, and some samples of Cheddar. In addition to the well-known low-galactose Swiss and Dutch cheeses, according to the current results, five Spanish cheeses can be safely consumed. The greater availability of types of cheese favours better bone health in patients with galactosaemia.

Patterns of Fourier-transform infrared estimated milk constituents in early lactation Holstein cows on a single New York State dairy.

Cows undergo immense physiological stress to produce milk during early lactation. Monitoring early lactation milk through Fourier-transform infrared (FTIR) spectroscopy might offer an understanding of which cows transition successfully. Daily patterns of milk constituents in early lactation have yet to be reported continuously, and the study objective was to initially describe these patterns for cows of varying parity groups from 3 through 10 d postpartum, piloted on a single dairy. We enrolled 1,024 Holstein cows from a commercial dairy farm in Cayuga County, New York, in an observational study, with a total of 306 parity 1 cows, 274 parity 2 cows, and 444 parity ≥3 cows. Cows were sampled once daily, Monday through Friday, via proportional milk samplers, and milk was stored at 4°C until analysis using FTIR. Estimated constituents included anhydrous lactose, true protein, and fat (g/100 g of milk); relative % (rel%) of total fatty acids (FA) and concentration (g/100 g of milk) of de novo, mixed, and preformed FA; individual fatty acids C16:0, C18:0, and C18:1 cis-9 (g/100 g of milk); milk urea nitrogen (MUN; mg/100 g of milk); and milk acetone (mACE), milk ß-hydroxybutyrate (mBHB), and milk-predicted blood nonesterified fatty acids (mpbNEFA) (all expressed in mmol/L). Differences between parity groups were assessed using repeated-measures ANOVA. Milk yield per milking differed over time between 3 and 10 DIM and averaged 8.7, 13.3, and 13.3 kg for parity 1, 2, and ≥3 cows, respectively. Parity differences were found for % anhydrous lactose, % fat, and preformed FA (g/100 g of milk). Parity differed across DIM for % true protein, de novo FA (rel% and g/100 g of milk), mixed FA (rel% and g/100 g of milk), preformed FA rel%, C16:0, C18:0, C18:1 cis-9, MUN, mACE, mBHB, and mpbNEFA. Parity 1 cows had less true protein and greater fat percentages than parity 2 and ≥3 cows (% true protein: 3.52, 3.76, 3.81; % fat: 5.55, 4.69, 4.95, for parity 1, 2, ≥3, respectively). De novo and mixed FA rel% were reduced and preformed FA rel% were increased in primiparous compared with parity 2 and ≥3 cows. The increase in preformed FA rel% in primiparous cows agreed with milk markers of energy deficit, such that mpbNEFA, mBHB, and mACE were greatest in parity 1 cows followed by parity ≥3 cows, with parity 2 cows having the lowest concentrations. When measuring milk constituents with FTIR, these results suggest it is critical to account for parity for the majority of estimated milk constituents. We acknowledge the limitation that this study was conducted on a single farm; however, if FTIR technology is to be used as a method of identifying cows maladapted to lactation, understanding variations in early lactation milk constituents is a crucial first step in the practical adoption of this technology.

Purification and Characterization of a Novel D-Galactose Binding Lectin from Seeds of Meizotropis buteiformis.

Plant lectins are carbohydrate-binding proteins that are ubiquitously found in almost all plant species and have different structures and functions depending on the sources. Purifying lectins from their plant sources and determining their sugar specificity become an important goal for evaluating their potential biomedical applications. Here, we report the affinity purification of a Dgalactose specific lectin from the seeds of Meizotropis buteiformis Voigt., and its physicochemical parameters, and LC-MS/MS (tandem mass spectrometry) analysis. Isolation and purification of this lectin were performed by simple successive steps of lectin extraction, ammonium sulphate fractionation, and affinity chromatography using lactose-linked Sepharose-4B chromatography column. The affinity-purified lectin has a native molecular weight of 75 kDa and is found to be a heterodimer (molecular weight of 36 and 38 kDa). The LC-MS/MS results suggested that the purified lectin had not been reported earlier. AIM: The main aim of the present study is to find out the novelty and characteristics of a lectin purified from the plant Meizotropis buteiformis. BACKGROUND: Lectins are proteins that possess the ability to specifically bind glycans of glycoconjugates. Plants are considered rich sources of lectins and the determination of sugar specificity of a purified plant lectin is an important aspect in order to evaluate its potential area of application. In the present study, a novel D-Galactose specific lectin is purified from Meizotropis buteiformis through affinity chromatography and examined for its various physical and biochemical characteristics. OBJECTIVE: The objective of the present study is to purify a novel lectin up to its homogeneity from the seeds of Meizotropis buteiformis and characterization of its various physical and biochemical properties. METHODS: The lectin was purified by simple successive steps of lectin extraction, ammonium sulphate fractionation, and affinity chromatography. Activity of the purified lectin was determined by hemagglutination assay. Some physicochemical parameters of the purified protein were also determined along with identification of protein by LC-MS/MS and the spectra analysis using Mascot sequence matching software (Matrix Science) with the NCBI database. RESULTS: From the current investigation, it was found that the purified lectin has a native molecular weight of 75 kDa. Among the various sugars and sugar derivatives tested, lactose and D-galactose were found to be potent inhibitors of its activity. Its optimum pH range was found to be from 6.5 to 7.5 and also it exhibited full activity at a temperature from 0ºC to 50ºC. The purified lectin does not show any effects on its activities for metal ions tested. The protein view report of the LC-MS/MS result analysis showed a 50% sequence similarity with that of the lectin beta-chain of the Butea monosperma. CONCLUSION: In the present study, a novel D-Galactose specific lectin is purified from Meizotropis buteiformis by affinity chromatography using Sepharose 4B. The purified lectin is found to be heterodimeric and metal ion independent. The LC-MS/MS results suggested that the purified lectin has not been reported earlier.

Miniaturized open-source 2LabsToGo screening of lactose-free dairy products and saccharide-containing foods.

The open-source 2LabsToGo system is the only one in its nature. It combines in one miniaturized instrument all relevant steps normally performed in a chemical and biological laboratory. For the first time, the applicability of the 2LabsToGo system was studied for screening 17 food products. As examples, saccharides were analyzed in eight products of different matrix complexity, and the absence of lactose was studied in nine lactose-free dairy products. Derivatization including homogeneous reagent application and plate heating via the 2LabsToGo system was explored for saccharide detection, and its performance was investigated. The visual detection sensitivity of lactose was comparable to previous studies. The precision of lactose in milk matrix (%RSD 4.6%) as well as the coefficient of determination of the calibration function (0.9995) were highly satisfying. The obtained lactose content of milk (4.5%) was plausible. Screening eight saccharide-containing food samples showed the saccharides in agreement with the expectations for the respective food product. The lactose content of nine different lactose-free dairy products was proven to be below the 0.1% lactose limit value. As proof-of-principle and for verification, these screening results obtained with the miniaturized 2LabsToGo system were reproduced using conventional state-of-the-art instrumentation, which led to the same results. However, instrumental costs were comparably low for the 2LabsToGo system. The application of the new 2LabsToGo system was successfully shown for saccharide screening, which is attractive to the field of quality control or official food control.

Spectroscopic analysis of the effect of glycation on casein structure and aggregation and its dependence on lactose concentration.

In order to study the effect of glycation on the molecular structure of casein (CN), three groups of simulated milk powders with different lactose (Lac) contents, CN:Lac = 1:1.5, 1:2, and 1:2.5 (m:m), were constructed. The glycation level and the structural changes of CN were analyzed by multiple spectra. The results showed that the interaction between Lac and CN molecules was based on the dynamic quenching of collision. A higher quenching constant, KSV, means that the 1:1.5 group showed more distinct glycation levels. In addition, CN glycation reduced the aggregation of casein micelles caused by heating treatment and repaired the structural damage of CN induced during the heating process. There was no positive correlation found between Lac content and protein glycation; thus, decreasing the Lac content of milk powder is not likely to be an effective method to prolong storage stability.

Diagnostic milk biomarkers for predicting the metabolic health status of dairy cattle during early lactation.

Data on metabolic profiles of blood sampled at d 3, 6, 9, and 21 in lactation from 117 lactations (99 cows) were used for unsupervised k-means clustering. Blood metabolic parameters included ß-hydroxybutyrate (BHB), nonesterified fatty acids, glucose, insulin-like growth factor-1 (IGF-1) and insulin. Clustering relied on the average and range of the 5 blood parameters of all 4 sampling days. The clusters were labeled as imbalanced (n = 42) and balanced (n = 72) metabolic status based on the values of the blood parameters. Various random forest models were built to predict the metabolic cluster of cows during early lactation from the milk composition. All the models were evaluated using a leave-group-out cross-validation, meaning data from a single cow were always present in either train or test data to avoid any data leakage. Features were either milk fatty acids (MFA) determined by gas chromatography (MFA [GC]) or features that could be determined during a routine dairy herd improvement (DHI) analysis, such as concentration of fat, protein, lactose, fat/protein ratio, urea, and somatic cell count (determined and reported routinely in DHI registrations), either or not in combination with MFA and BHB determined by mid-infrared (MIR), denoted as MFA [MIR] and BHB [MIR], respectively, which are routinely analyzed but not routinely reported in DHI registrations yet. Models solely based on fat, protein, lactose, fat/protein ratio, urea and somatic cell count (i.e., DHI model) were characterized by the lowest predictive performance [area under the receiver operating characteristic curve (AUCROC) = 0.69]. The combination of the features of the DHI model with BHB [MIR] and MFA [MIR] powerfully increased the predictive performance (AUCROC = 0.81). The model based on the detailed MFA profile determined by GC analysis did not outperform (AUCROC = 0.81) the model using the DHI-features in combination with BHB [MIR] and MFA [MIR]. Predictions solely based on samples at d 3 were characterized by lower performance (AUCROC DHI + BHB [MIR] + MFA [MIR] model at d 3: 0.75; AUCROC MFA [GC] model at d 3: 0.73). High predictive performance was found using samples from d 9 and 21. To conclude, overall, the DHI + BHB [MIR] + MFA [MIR] model allowed to predict metabolic status during early lactation. Accordingly, these parameters show potential for routine prediction of metabolic status.