Impact of Processing Methods on the Distribution of Mineral Elements in Goat Milk Fractions.

Milk and dairy products are excellent sources of mineral elements, including Ca, P, Mg, Na, K and Zn. The purpose of this study was to determine the effect of non-thermal (homogenization) and thermal (heat treatment) treatments on the distribution of mineral elements in 4 milk fractions: fat, casein, whey protein, and aqueous phase. The study results revealed that the distribution of mineral elements (such as Mg and Fe) in fat fractions is extremely low, while significant mineral elements such as Ca, Zn, Fe, and Cu are mostly dispersed in casein fractions. For non-treated goat milk, Mo is the only element identified in the whey protein fraction, while K and Na are mostly found in the aqueous phase. Mineral element concentrations in fat (K, Zn, etc.) and casein fraction (Fe, Mo, etc.) increased dramatically after homogenization. Homogenization greatly decreased the concentration of mineral elements in the whey protein fraction (Ca, Na, etc.) and aqueous phase (Fe, Cu, etc.). After heat treatment, the element content in the fat fraction and casein fraction increased greatly when compared with raw milk, such as Cu and Mg in the fat fraction, Na and Cu in the whey protein fraction, the concentration of components such as Mg and Na in casein fraction increased considerably. On the other hand, after homogenization, Zn in the aqueous phase decreased substantially, whereas Fe increased significantly. Therefore, both homogenization and heat treatment have an effect on the mineral element distribution in goat milk fractions.

Effect of lactation on the distribution of mineral elements in goat milk.

The distribution of mineral elements in milk is crucial for their absorption and utilization, however, there has been limited attention given to the status of mineral elements in goat milk. In this study, goat milk was collected at 4 lactation periods (1-3, 90, 150, 240 d) and separated into 4 fractions (fat, casein, whey, and aqueous phase). The concentrations of Mg, Ca, Na, K, Zn, Fe, Cu, Mn, Co, Ni, Mo, and Cr in 4 fractions were analyzed using an inductively coupled plasma emission spectrometer. Our findings reveal that Ca, Zn, Fe, Cu, Mn, and Cr exhibit the highest levels in casein, while Mo demonstrates the highest content in whey. Additionally, Mg, Na, K, and Ni display the highest concentrations in the aqueous phase. Specifically, the contents of Ca, Cu and Fe in casein decrease from 1-3 to 150 d of lactation but increase from 150 to 240 d of lactation. Furthermore, the content of Mg in the aqueous phase decreases from 1-3 to 90 d of lactation but increases from 90 to 240 d of lactation. The content of Na and K in the aqueous phase decreases from 1-3 to 150 d of lactation. Notably, the content of Mo in whey increases from 1-3 to 150 d of lactation and decreases from 150 to 240 d. Our research contributes to the advancement of understanding the bioavailability of mineral elements in goat milk.

Identification markers of goat milk adulterated with bovine milk based on proteomics and metabolomics.

This study investigated the differences in proteins and metabolites from goat and bovine milk, and their mixtures, using data-independent-acquisition-based proteomics and metabolomics methods. In the skim milk, relative abundances of secretoglobin family 1D member (SCGB1D), polymeric immunoglobulin receptor, and glycosylation-dependent cell adhesion molecule 1 were increased, with an increase in the amount of 1-100 % bovine milk and served as markers at the 1 % adulteration level. In whey samples, ß-lactoglobulin and α-2-HS-glycoprotein could be used to detect adulteration at the 0.1 % adulteration level, and SCGB1D and zinc-alpha-2-glycoprotein at the 1 % level. The metabolites of uric acid and N-formylkynurenine could be used to detect bovine milk at adulteration levels as low as 1 % based on variable importance at a projection value of > 1.0 and P-value of < 0.05. Our findings suggest novel markers of SCGB1D, uric acid, and N-formylkynurenine that can help to facilitate assessments of goat milk authenticity.

Comparative analysis of changes in whey proteins of goat milk throughout the lactation cycle using quantitative proteomics.

The composition and content of goat milk proteins are affected by many factors and have been extensively studied. However, variation in whey protein composition in goat milk throughout the lactation cycle has not been clarified. In the current study, 15 dairy goats were selected, and milk samples were collected at 1, 3, 30, 90, 150, and 240 d after delivery. Whey proteins were separated and digested and then identified using data-independent acquisition (DIA) and data-dependent acquisition proteomics approaches. Protein profiles identified using DIA were consistent with those of the data-dependent acquisition proteomics approach according to clustering and principal component analyses. Significant differences in the abundance of 238 proteins around the lactation cycle were identified using the DIA approach. Developmental changes of the whey proteome corresponding to lactation stage were revealed: plasminogen, α-2-macroglobulin, and fibronectin levels decreased from d 1 to 240, whereas polymeric immunoglobulin receptor, nucleobindin 2, fatty acid-binding protein 3, and lactoperoxidase increased from d 1 to 240. Protein-protein interaction analysis showed that fibronectin with a higher degree of connectivity is a central node. The findings are of great significance to better understanding the potential role of specific proteins and the mechanism of protein biosynthesis or intercellular transport in the mammary glands related to the physiological changes of dairy goats.

Developmental changes in proteins of casein micelles in goat milk using data-independent acquisition-based proteomics methods during the lactation cycle.

Casein micelles (CM) play an important role in milk secretion, stability, and processing. The composition and content of milk proteins are affected by physiological factors, which have been widely investigated. However, the variation in CM proteins in goat milk throughout the lactation cycle has yet to be fully clarified. In the current study, milk samples were collected at d 1, 3, 30, 90, 150, and 240 of lactation from 15 dairy goats. The size of CM was determined using laser light scattering, and CM proteins were separated, digested, and identified using data-independent acquisition (DIA) and data-dependent acquisition (DDA)-based proteomics approaches. According to clustering and principal component analysis, protein profiles identified using DIA were similar to those identified using the DDA approach. Significant differences in the abundance of 115 proteins during the lactation cycle were identified using the DIA approach. Developmental changes in the CM proteome corresponding to lactation stages were revealed: levels of lecithin cholesterol acyltransferase, folate receptor α, and prominin 2 increased from 1 to 240 d, whereas levels of growth/differentiation factor 8, peptidoglycan-recognition protein, and 45 kDa calcium-binding protein decreased in the same period. In addition, lipoprotein lipase, glycoprotein IIIb, and α-lactalbumin levels increased from 1 to 90 d and then decreased to 240 d, which is consistent with the change in CM size. Protein-protein interaction analysis showed that fibronectin, albumin, and apolipoprotein E interacted more with other proteins at the central node. These findings indicate that changes in the CM proteome during lactation could be related to requirements of newborn development, as well as mammary gland development, and may thus contribute to elucidating the physical and chemical properties of CM.

Distribution and variation in proteins of casein micellar fractions response to heat-treatment from five dairy species.

Casein micelles (CMs) contribute to the physicochemical properties and stability of milk. However, how the proteome of CMs changes following heat treatment has not been elucidated. Here, changes in the proteins of CMs in samples of Holstein, buffalo, yak, goat, and camel milk following heat treatment were investigated using a LC-MS/MS approach. According to the hierarchical clustering results, Holstein, yak, and buffalo milk samples had similar CMs protein components, followed by goat and camel milk samples. Changes in lipoprotein lipase and α-lactalbumin in CMs were dependent on the intensity of heat treatment and were similar among the studied species, whereas changes in κ-casein, lactoferrin, and apolipoprotein A-I differed among different types of milk. These results provide information on the distribution and variations of the proteomes of CMs following heat treatment, which will assist in the identification of proteins that are dissociated and attached to CMs from different dairy species during heat treatment.

Prevalence, Drug Resistance, and Virulence Genes of Potential Pathogenic Bacteria in Pasteurized Milk of Chinese Fresh Milk Bar.

Fresh Milk Bar (FMB), an emerging dairy retail franchise, is used to instantly produce and sell pasteurized milk and other dairy products in China. However, the quality and safety of pasteurized milk in FMB have received little attention. The objective of this study was to investigate the prevalence, antimicrobial resistance, and virulence genes of Escherichia coli, Staphylococcus aureus, and Streptococcus in 205 pasteurized milk samples collected from FMBs in China. Four (2.0%) isolates of E. coli, seven (3.4%) isolates of S. aureus, and three (1.5%) isolates of Streptococcus agalactiae were isolated and identified. The E. coli isolates were resistant to amikacin (100%), streptomycin (50%), and tetracycline (50%). Their detected resistance genes include aac(3)-III (75%), blaTEM (25%), aadA (25%), aac(3)-II (25%), catI (25%), and qnrB (25%). The S. aureus isolates were mainly resistant to penicillin G (71.4%), trimethoprim-sulfamethoxazole (71.4%), kanamycin (57.1%), gentamicin (57.1%), amikacin (57.1%), and clindamycin (57.1%). blaZ (42.9%), mecA (28.6%), ermB (14.3%), and ermC (14.3%) were detected as their resistance genes. The Streptococcus strains were mainly resistant to tetracycline (66.7%) and contained the resistance genes pbp2b (33.3%) and tetM (33.3%). The virulence genes eae and stx2 were only found in one E. coli strain (25%), sec was detected in two S. aureus strains (28.6%), and bca was detected in one S. agalactiae strain (33.3%). The results of this study indicate that bacteria with drug resistance and virulence genes isolated from the pasteurized milk of FMB are a potential risk to consumers' health.

Occurrence, Antimicrobial Resistance Patterns, and Genetic Characterization of Staphylococcus aureus Isolated from Raw Milk in the Dairy Farms over Two Seasons in China.

This study investigated the occurrence and resistance rates of Staphylococcus aureus strains isolated from raw milk in the dairy farms over two seasons (spring and autumn) and across four regions that included 11 provinces in China. In total, 750 raw milk samples from the 405 dairy farms were collected. Fifteen antimicrobial agents were tested for antimicrobial resistance via disk diffusion tests, and PCR tests were performed to identify drug resistance genes of S. aureus isolates. Out of 750 samples, 276 (36.8%) were positive for S. aureus, with 150 (41.1%) being positive in spring and 126 (32.7%) being positive in autumn. The occurrence rate of S. aureus in northeastern China (45%) was higher than that in western China (33%) and southern China (31.9%), respectively, and the rate significantly (p < 0.05) differed from those of western China and southern China. Of 276 isolates, 261 (94.6%) strains were resistant to more than 1 antimicrobial drug, and 193 (69.9%) strains were multidrug resistant. The blaZ (46.3%), dfrG (35.5%), and tetM (27.2%) genes were detected at a high frequency in the S. aureus strains. Our data revealed a variation (p < 0.05) in the resistance patterns in the different regions and across the two seasons. The occurrence and drug resistance rates of S. aureus isolated from raw milk obtained from dairy farms may still cause severe problems in China.

Survey of Aflatoxin M1 in Commercial Liquid Milk Products in China.

ABSTRACT: The objective of this study was to investigate the occurrence of aflatoxin M1 (AFM1) in Chinese liquid milk products. A total of 190 liquid milk samples, including 168 ultrahigh-temperature-treated milk samples and 22 pasteurized milk (PM) samples, were collected in August 2019. A screening assay with the Charm rapid test kit and a confirmation method with high-performance liquid chromatography were used for AFM1 analysis. Nine (4.74%) samples were screened positive, of which 5 (2.11%) samples were confirmed with concentration levels of 0.022 to 0.049 µg/kg. The AFM1 levels confirmed were all below the maximum residue levels set by China, the European Union, the United States, and the Codex Alimentarius Commission. The detection rate of AFM1 in domestic milk samples was 3.39%, while no AFM1 was detected in samples of imported milk. The prevalences of AFM1 detected in three groups of brands were as follows: group I, the major brands of China, 2.70%; group II, the local city domestic brands, 4.55%; and group III, the brands imported into China, 0. The detection rate of AFM1 was significantly higher in PM samples (9.09%) than in ultrahigh-temperature-treated samples (1.19%) (P < 0.05). Although the residue level of AFM1 did not exceed the maximum residue levels in any of the samples, the higher detection rate in local Chinese brands, especially in PM samples, deserves the attention of the Chinese government and consumers.

Comparative proteomic characterization of bovine milk containing ß-casein variants A1A1 and A2A2, and their heterozygote A1A2.

BACKGROUND: Genetic variants of ß-casein are cosnidered to affect the components of milk. However, limited data are available on the bovine protein components correlated with ß-casein variants at the proteome level. In the present study, cows producing milk containing ß-casein variants (A1A1 and A2A2) and their heterozygote (A1A2) were identified using a high-resolution melting method, and milk samples were collected and tested. Comparative analyses of casein micelles, whey and milk fat globule membrane fractions in each milk variant were performed using a label-free proteomics approach. RESULTS: The results obtained showed that ceruloplasmin and cathelicidin-2 were the most abundant proteins in milk containing variant A1A1; lactoferrin and CD5 molecule-like were the most abundant proteins in milk containing variant A2A2; and selenoprotein P and osteopontin were the most abundant proteins in milk containing heterozygote A1A2. Differences in protein components in milk containing the different ß-casein variants were visualized using hierarchical clustering, and profiles were separated using principal components analysis. The differentially expressed proteins in milk containing A1A1, A2A2 or A1A2 were predominantly involved in response to stress and defense response according to their Gene Ontology annotations. CONCLUSION: Our findings provide new insights into differentially expressed milk proteins corresponding to the presence of different ß-casein variants. This knowledge will help determine their potential biological functions in dairy products and the effects on human health. © 2020 Society of Chemical Industry.

A Survey of 61 Veterinary Drug Residues in Commercial Liquid Milk Products in China.

ABSTRACT: To investigate the drug residue status in commercial liquid milk products in China, 190 samples, including ultrahigh temperature milk (n = 168) and pasteurized milk (n = 22) samples, were collected in 2019. Milk samples were analyzed for the presence of any of the 61 veterinary drugs in them by using a screening assay combined with an ultraperformance liquid chromatography-tandem mass spectrometry analysis. Ten (5.26%) samples were found positive for ß-lactams, tetracyclines, and aminoglycosides, and six (3.16%) samples were confirmed residual for penicillin G (n = 6; 3.16%), tetracycline (n = 1; 0.53%), and oxytetracycline (n = 1; 0.53%), with the maximum concentration of 2.85, 40.64, and 12.35 µg kg-1, respectively. Drug residue detection rate in group II (4.55%; the local city domestic brands) was higher than that in group I (2.70%; the major brands of China) and group III (2.78%; the imported brands into China) and higher in domestic samples (3.39%) than that in imported samples (2.78%), and higher in pasteurized milk samples (9.09%) than in ultrahigh temperature milk samples (2.38%). All drug residue levels were far below the regulated maximum residue limits. However, based on some veterinary drug residues detected in the samples, there is a potential veterinary drug risk in liquid milk products in the Chinese market, and this situation deserves the attention of governments and consumers.

Analysis of Veterinary Drug Residues in Pasteurized Milk Samples in Chinese Milk Bars.

ABSTRACT: The milk bar is an emerging style of retail business that mainly produces pasteurized milk (PM) and other dairy products on-site in many large cities of the People's Republic of China. To date, no data about veterinary drug residues in PM samples produced from milk bars have been reported. The objective of this study was to investigate the safety of PM from a total of 182 PM samples collected from milk bars from 10 provincial capital cities and to analyze the residues of seven classes of 61 veterinary drugs. First, the chemical components were screened with test kits, and then the positive samples were further confirmed by ultraperformance liquid chromatography-tandem mass spectrometry. The results showed that 15 (8.24%) samples were screened positive for veterinary drugs, and six drugs in 11 (6.04%) samples were confirmed. The veterinary drugs detected were penicillin G (2.20%), tetracycline (1.10%), tylosin (1.10%), amoxicillin (0.55%), oxytetracycline (0.55%), and gentamicin (0.55%), with maximum residue levels of 3.4, 11.9, 28.2, 3.0, 26.9, and 63.5 µg kg-1, respectively. Veterinary drug residues were detected as positive in 7 of 10 cities, with the highest detection rate as 14.29% in Urumqi. No positive samples were found in the cities of Nanjing, Tianjin, and Nanning. All detected drug levels were far below the maximum residue levels regulated by China, the European Union, and the Codex Alimentarius Commission. This suggests that the overall veterinary drug residues in PM in milk bars reached the safety code of the country. However, potential risks still exist, and continuous attention should be paid to guarantee the safety of this milk product in the future.

Elimination kinetics of ceftiofur hydrochloride in milk after an 8-day extended intramammary administration in healthy and infected cows.

Ceftiofur hydrochloride (CEF) is occasionally used for the intramammary (IMM) treatment of mastitis. This extralabel manner could result in a drug-residue violation of the milk. The objective of this study was to determine the elimination kinetics of IMM CEF in lactating dairy cattle. The pharmacokinetic profile of CEF after repeated IMM administration in nine healthy cows and nine Staphylococcus aureus infected cows was investigated, alongside determining the MICs of Staph. aureus field strains. The MIC 90 value for CEF in Staph. aureus field strains (n = 31) was 0.25 µg/mL. The t >MIC CEF values for low- production quarters were longer than those for high- and mid- production quarters. The results showed that ceftiofur was detected in milk up to 108 h after the last infusion in both healthy and infected cows. Cows with low milk production eliminate IMM drugs more slowly than cows with higher production. Our findings suggest that this extralabel use is not encouraged and a prudent use is recommended for mastitis therapy. The use of CEF should be reserved for infections where susceptibility tests indicate its efficacy and when alternatives are not available.

Survey of Veterinary Drug Residues in Raw Milk in Hebei Province, China.

The objective of this study was to investigate the occurrence of veterinary drug residues in raw milk from Hebei, the second-largest dairy production province in the People's Republic of China. A total of 192 raw milk samples were collected from 64 milk stations in seven districts. Twenty-eight veterinary drug residues were analyzed by ultraperformance liquid chromatography with tandem mass spectrometry based on a China National Standard. Raw milk samples with multiple residues of veterinary drugs were not found in the present study. Residues of four veterinary drugs, penicillin G, sulfacetamide, trimethoprim, and lincomycin, were detected in 12 (6.25%) raw milk samples, with detection ratios of 1.04, 0.52, 3.13, and 1.56%, respectively. All veterinary drug residues detected were under the maximum residue levels as regulated by China, the European Union, the United States, and the Codex Alimentarius Commission. In general, raw milk from Hebei province was considered relatively safe for human consumption because of the low prevalence of veterinary drug residues. However, stringent control measurements for veterinary drug residues in raw milk are required because some veterinary drugs were detected in milk from some areas of Hebei province.