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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.

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 t...

BACKGROUND Genetic variants of β-casein are cosnidered to affect the components of milk. However, limited data are available on the bovine protein components correlated with β-casein variants at the proteome level. In the present study, cows producing milk containing β-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 β-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 β-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.

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.

The study was conducted to investigate the effects of protein supplement (soybean meal and wheat bran) on the growth performance and blood parameters of native lamb. A total number of twelve indigenous lambs aged 10 months were randomly distributed into four dietary groups having 3 replications. First group (T1) was fed control diet, second group (T2) was provided 50 g protein supplement + green grass, third group (T3) was provided 75 g protein supplement + green grass and fourth group (T4) was fed 100 g protein supplement + green grass. The results showed that body weight gain was similarly significantly higher (P

This study was aimed to evaluate the effects of using different levels of olive (Olea europaea) leaf extract on fresh and preserved mutton meatballs. Meatballs were divided into four different groups and treated as T0 (0), T1 (0.1), T2 (0.2) and T3 (0.3%), respectively based on olive leaf extract supplementation. Days of intervals of experiment were 0, 5, 10 days. Samples were preserved at 4˚C for up to 10 days. Different types of analysis such as, sensory (color, flavor, juiciness and overall acceptability), proximate composition (CP %), physicochemical (pH), biochemical (POV, FFA and TBARS) and microbiological (TVC, TCC and TYMC) were determined. Color, flavor and acceptability reduced significantly (p

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. HIGHLIGHTS

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.

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.

Objectives: The objective was to find the differences in the prevalence and resistance of Escherischia coli isolated from raw milk samples from mastitis cases in four regions of China. Methods: A total of 750 bovine raw milk samples from mastitis cases were collected from four regions of China over two seasons. Antimicrobial resistance against 29 antimicrobial agents was determined, and 27 drug-resistant genes were tested. Results: Eighty-three strains (11.1%) of E. coli were isolated and identified. No significant differences in the number of E. coli isolates were observed between the two sampling seasons in the same regions (P > 0.05). However, a significant difference in E. coli prevalence was found among the four different regions (P

Streptococcus is a major mastitis-causing pathogen in dairy cows. To investigate the prevalence, antimicrobial resistance and virulence gene of Streptococcus in mastitic milk, a total of 735 mastitic raw milk samples from dairy cows in 11 provinces of China were collected and tested. Antimicrobial resistance of Streptococcus isolates was determined by disc diffusion against 8 classes 29 antimicrobial agents, and Streptococcus resistant genes and virulence genes were determined by PCR and agarose gel electrophoresis. A total of 64 (8.71%) isolates of Streptococcus were isolated and identified using biochemical profiling, including 22 isolates of Streptococcus agalactiae, 13 isolates of Streptococcus dysgalactiae, and 29 isolates of Streptococcus uberis. Out of 64 resistant Streptococcus isolates, all isolates (100%) were resistant to 3 or more antimicrobials. The most frequency (n = 18, 28.12%) of the isolates were multi-resistant to 5–7 antimicrobials and the highest multi-resistant number was 29 (n = 1, 1.56%). Streptococcus isolates had the highest resistance rate to tetracycline (98.44%) and oxacillin (98.44%), followed by penicillin G (96.88%) and doxycycline (96.88%), and the lowest resistance was observed with respect to ciprofloxacin (1.56%). A total of 16 antimicrobials resistance genes with 25 combination patterns were detected in the isolates. The gene combination of Sul1/Sul2/Sul3 + gyrA/parC + cat1/cat2 was the most common pattern (12.5%). The correlation between resistant phenotypes and resistance genes in Streptococcs was 35.87%. A total of 7 virulence genes were detected and 59 (92.19%) isolates harbored at least one gene. Twenty-four classes of gene patterns were found in the isolates and the patterns of bca (9.38%) and cfb (9.38%) were the most prevalent form. In conclusion, the issue of drug resistance of Streptococcus is still a great concern in cattle health in China.

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.

A selective and rapid method has been developed to determine, simultaneously, 38 veterinary antibiotic residues in raw milk by ultra-high-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). One milliliter of raw milk was diluted with 0.5 mL water and 3 mL acetonitrile, then purified using an Oasis HLB cartridge. The eluates were evaporated by nitrogen drying and then reconstituted to 4 mL with water/acetonitrile (8:1) before being injected into the UPLC–MS/MS system. The results indicated recoveries of 68–118% for 14 β-lactams, 79–118% for eight quinolones, 71–106% for eight sulfonamides, 76–116% for four tetracyclines, 78–106% for three macrolides, and 88–103% for one lincosamides, with coefficients of variation less than 15% for intraday and interday precisions. The limit of quantification for all antibiotics was 0.03–10 μg kg −1 . This methodology was then applied to field-collected real raw milk samples and trace levels of four antibiotics were detected.