Short communication: Space allocation in intensive Mediterranean buffalo production influences the profile of functional biomolecules in milk and dairy products

The aim of the present study was to determine if space allocation influenced the concentration of biomolecules in buffalo milk and dairy products. Intensively housed buffaloes (n = 96) were randomly assigned to 2 groups according to days in milk, parity, and milk yield: group S10 had a space allocation of 10 m² per buffalo and group S15 had a space allocation of 15 m² per buffalo. Individual milk yield was recorded daily. Twice a month, a bulk milk sample was collected for each group, as well as whey, ricotta, and mozzarella cheese, to assess cheese yield and to conduct HPLC-electrospray ionization-tandem mass spectrometry, milk antioxidant activity, and cell viability analyses. We tested milk extracts from the 2 groups in vitro to evaluate their efficacy in counteracting endothelial oxidative damage induced by high glucose. We evaluated reproductive function in 28 buffaloes from each group using the Ovsynch-timed artificial insemination program. We observed no differences in milk quantity or quality in terms of fat, protein, or lactose, and reproductive function did not differ between the 2 groups. Compared with group S10, group S15 had higher concentrations of carnitine (56.7 ± 1.1 vs. 39.8 ± 0.7 mg/L in milk and 40.9 ± 0.8 vs. 31.7 ± 0.7 mg/L in whey), acetyl-l-carnitine (51.9 ± 0.3 vs. 39.7 ± 0.7 mg/L in milk and 41.1 ± 1.7 vs. 28.7 ± 2.6 mg/L in whey), propionyl-l-carnitine (34.8 ± 1.0 vs. 21.0 ± 0.9 mg/L in milk and 26.9 ± 0.8 vs. 17.6 ± 1.2 mg/L in whey), glycine betaine (23.1 ± 1.9 vs. 13.5 ± 1.6 mg/L in milk and 10.7 ± 0.4 vs. 7.9 ± 0.5 mg/L in whey), and δ-valerobetaine (24.2 ± 0.5 vs. 16.7 ± 0.5 mg/L in milk and 22.0 ± 0.9 vs. 15.5 ± 0.7 mg/L in whey). Group S15 also had higher total antioxidant activity than group S10 (56.7 ± 1.9 vs. 46.4 ± 1.13 mM Trolox equivalents). Co-incubation of high-glucose-treated endothelial cells with milk extracts from group S15 improved cell viability compared with cells treated with high glucose only; it also reduced intracellular lipid peroxidation (144.3 ± 0.4 vs. 177.5 ± 1.9%), reactive oxygen species (141.3 ± 0.9 vs. 189.3 ± 4.7 optical density units), and cytokine release (tumor necrosis factor-α, IL-1β, IL-6). Greater space allocation was associated with higher levels of biomolecules in buffalo milk. This could have been the result of improved welfare in buffaloes that were allocated more space.     

Short communication: Prediction of milk coagulation and acidity traits in Mediterranean buffalo milk using Fourier-transform mid-infrared spectroscopy

Milk coagulation and acidity traits are important factors to inform the cheesemaking process. Those traits have been deeply studied in bovine milk, whereas scarce information is available for buffalo milk. However, the dairy industry is interested in a method to determine milk coagulation and acidity features quickly and in a cost-effective manner, which could be provided by Fourier-transform mid-infrared (FT-MIR) spectroscopy. The aim of this study was to evaluate the potential of FT-MIR to predict coagulation and acidity traits of Mediterranean buffalo milk. A total of 654 records from 36 herds located in central Italy with information on milk yield, somatic cell score, milk chemical composition, milk acidity [pH, titratable acidity (TA)], and milk coagulation properties (rennet coagulation time, curd firming time, and curd firmness) were available for statistical analysis. Reference measures of milk acidity and coagulation properties were matched with milk spectral information, and FT-MIR prediction models were built using partial least squares regression. The data set was divided into a calibration set (75%) and a validation set (25%). The capacity of FT-MIR spectroscopy to correctly classify milk samples based on their renneting ability was evaluated by a canonical discriminant analysis. Average values for milk coagulation traits were 13.32 min, 3.24 min, and 39.27 mm for rennet coagulation time, curd firming time, and curd firmness, respectively. Milk acidity traits averaged 6.66 (pH) and 7.22 Soxhlet-Henkel degrees/100 mL (TA). All milk coagulation and acidity traits, except for pH, had high variability (17 to 46%). Prediction models of coagulation traits were moderately to scarcely accurate, whereas the coefficients of determination of external validation were 0.76 and 0.66 for pH and TA, respectively. Canonical discriminant analysis indicated that information on milk coagulating ability is present in the MIR spectra, and the model correctly classified as noncoagulating the 91.57 and 67.86% of milk samples in the calibration and validation sets, respectively. In conclusion, our results can be relevant to the dairy industry to classify buffalo milk samples before processing.     

Detection of buffalo milk adulteration with cow milk by capillary electrophoresis analysis

The addition of cow milk during the production of buffalo mozzarella is a common fraud in dairy industries because of the lower price and greater availability of cow milk throughout the year. The aim of this study was to develop a new, rapid, and robust capillary electrophoresis method for detecting and quantifying cow milk in buffalo milk by exploiting cow α-lactalbumin as a marker of adulteration. In particular, a linear calibration curve was generated, using a training set of calibrators consisting of 7 series of 17 buffalo/bovine whey mixtures, obtained after casein precipitation, with increasing percentages of cow whey. The capillary electrophoresis method showed high linearity (R² = 0.968), repeatability [relative standard deviation (RSD) = 2.11, 3.02, 4.38, and 1.18%, respectively for 5, 10, 20, and 50% of buffalo/bovine whey mixtures], and intermediate precision (RSD = 2.18, 2.49, 5.09, and 3.19%, respectively, for 5, 10, 20, and 50% buffalo/bovine whey mixtures). Moreover, the minimum amount of detectable fraudulent cow milk was 1%, and the limit of quantification was 3.1%.     

Short communication: Phenotypic characterization of total antioxidant activity of buffalo,goat, and sheep milk

Free radicals are reactive and unstable waste molecules produced by cells, responsible of damages and alteration on DNA, proteins, and fat. The daily intake of antioxidant compounds, acting against free radicals and their detrimental effects, is essential for human health. Milk contains several compounds with antioxidant activity, and the sum of their reducing potential blocking free radicals development is defined as total antioxidant activity (TAA). This novel trait has been described in literature both in individual and bulk cow milk, but there are no reports from other dairy species. Therefore, the present study aimed to investigate phenotypic variation of TAA in individual samples of buffalo (n = 105), goat (n = 112), and sheep (n = 198) milk. Total antioxidant activity was measured through a reference spectrophotometric method, and expressed as millimoles per liter of Trolox equivalents (TE). The greatest TAA was observed in sheep milk, averaging 7.78 mmol/L of TE and showing also the broadest phenotypic variation expressed as coefficient of variation (13.98%). Significantly lower TAA values were observed for buffalo (7.35 mmol/L of TE) and goat (6.80 mmol/L of TE) milk, with coefficients of variation of 8.18 and 8.47%, respectively. Total antioxidant activity exhibited weak correlations with milk yield and chemical composition. Phenotypic values of TAA presented in this study will be used to assess the ability of mid-infrared spectroscopy to predict this new trait and thus to collect data at the population level.     

Short communication: Stabilization of milk proteins at pH 5.5 using pectic polysaccharides derived from potato tubers

Potato pectin has unique molecular characteristics that differentiate it from commercially available pectins sourced from citrus peels or apple pomace, including a higher degree of branching and a higher acetyl content. The objective of this study was to evaluate the ability of potato pectin to stabilize milk proteins at an acidic pH above their isoelectric point, pH 5.5, at which no citrus- or apple-derived pectins are functional. Potato pectin was extracted from raw potato tubers by heating at pH 4.5 and 120°C for 30 min after removing starch solubilized using a dilute HCl solution adjusted to pH 2. The potato pectin was found to have a galacturonic acid content of 17.31 ± 3.29% (wt/wt) and a degree of acetylation of 20.20 ± 0.12%. A portion of the potato pectin was deacetylated by heating it in an alkaline condition. The deacetylation resulted in a galacturonic acid content of 19.12 ± 4.64% (wt/wt) and a degree of acetylation of 3.03 ± 0.03%. Particle size distributions in acidified milk drink (AMD) samples adjusted to pH 5.5 demonstrated that the acetylated and deacetylated potato pectins were capable of inhibiting the aggregation of milk proteins to the largest degree at a pectin concentration of 1.0 and 0.25% (wt/wt), respectively. Pectin molecules that were not bound to milk proteins in these AMD samples were quantified after centrifugally separating milk proteins and pectin bound to them from the serum. We found that, for the acetylated and deacetylated potato pectins, all or approximately half of the pectin molecules were bound to milk proteins at a pectin concentration of 0.25 or 1.0% (wt/wt), respectively. These results suggest that the presence of acetyl groups is a critical factor that determines how potato pectin molecules bind electrostatically to milk protein surfaces, form 3-dimensional structures there, and function as a stabilizer. The present results demonstrate that potato pectin can stabilize milk proteins at pH 5.5 and potentially enable the development of novel AMD products with improved functionality for casein-containing products with moderately acidic pH profiles.     

Short communication: Detection of undeclared presence of bovine milk in buffalo yogurt

The authenticity of buffalo (Bubalus bubalis) dairy products is a focal issue, considering the increasing demand for buffalo milk products. Therefore, the aim of this study was to investigate the undeclared presence of bovine (Bos taurus) milk in buffalo yogurt, to understand which risk factors might make the product vulnerable to fraud. Real-time PCR assay showed the undeclared presence of bovine DNA in addition to buffalo DNA in 18 of 72 samples. Given the widespread lack of data on the presence of undeclared milk species in buffalo dairy products, the study provides a significant insight into the incidence of fraud in the buffalo dairy field. The data from this study could help improve the analysis of food safety risks along the buffalo milk supply chain and in the dairy processing industry, perceived as being highly vulnerable to food fraud, and prioritize target areas for food policy making to steer and enforce European food fraud regulations.     

Short communication: Parapoxvirus and Orthopoxvirus coinfection in milk of naturally infected cows

Several studies have shown the occurrence of poxvirus infections associated with exanthematic lesions in cattle from many Brazilian states. Coinfection between viruses belonging to 2 genera, Orthopoxvirus (OPXV) and Parapoxvirus (PPV), was already identified from the lesions of affected cows and humans. The DNA and infectious viral particles of Vaccinia virus, an OPXV, have been detected in milk of naturally and experimentally infected cows. However, to date no reports have described the detection of Pseudocowpox virus, a PPV, in milk. Thus, we investigated the presence of PPV and OPXV in milk samples obtained from dairy cows from a Brazilian region with exanthematic disease outbreaks. From 2011 to 2014, 6 dairy farms with exanthematic disease outbreaks involving dairy cows, calves, and humans were visited. Twelve crusts of cows' teat lesions and 60 milk samples were collected. The crusts and milk samples were analyzed by PCR to detect OPXV or PPV DNA. According to the analyzed crusts, we detected PPV infection in 4 of the 6 visited farms, from which we investigated the PPV contamination in milk. From the 40 milk samples tested, PPV DNA was detected in 12 samples. Of these milk samples, 8 were positive for both PPV and OPXV. This is the first report of PPV DNA detection in milk samples from affected cows, indicating that the virus may be present in milk and potentially contaminating dairy products associated or not with OPXV. In addition to the lesions caused by direct contact, the presence of 2 or more poxvirus species in milk showed that the effect of zoonotic exanthematic diseases on public health and animal husbandry is relevant and cannot be overlooked.     

Short communication: Physicochemical features and microbial community of milk kefir using a potential probiotic Saccharomyces cerevisiae KU200284

The aim of this study was to analyze the β-glucan contents, physicochemical features, and microbial communities in milk kefir prepared using Saccharomyces cerevisiae KU200284 isolated from cucumber jangajji, a fermented vegetable commonly eaten in Korean. Three types of milk kefir were manufactured, with (1) activated kefir grain, (2) activated kefir grain with commercial S. cerevisiae BOF, and (3) activated kefir grain with S. cerevisiae KU200284. β-Glucan contents of milk kefir using kefir grain and kefir grain with S. cerevisiae strains BOF and KU200284 were 8.29, 8.59, and 8.57%, respectively. The pH, titratable acidity, viscosity, Brix level, and alcohol contents of milk kefir using kefir grain with S. cerevisiae strains were acceptable compared with milk kefir using only kefir grain. In milk kefir produced using kefir grains and S. cerevisiae strains, 16S rRNA reads showed representative strains of Lactobacillus kefiranofaciens (>72% relative abundance) and Acetobacter fabarum (>16% relative abundance). In particular, milk kefir using kefir grain with S. cerevisiae KU200284 had the highest relative abundance of L. kefiranofaciens. In addition, the internal transcribed sequence (ITS) rRNA reads in tested milk kefir showed representative strains of Kluyveromyces marxianus (>52% relative abundance) and Saccharomyces cerevisiae (>16% relative abundance). In contrast, milk kefir using S. cerevisiae strains had higher relative abundance of S. cerevisiae (>37%). The β-glucan production, physicochemical properties, and microbial community profiling indicate that S. cerevisiae KU200284 could be used in functional dairy products as a starter culture.     

Effect of intramammary infection with non-aureus staphylococci in early lactation in dairy heifers on quarter somatic cell count and quarter milk yield during the first 4 months of lactation

A longitudinal study was conducted to assess to what extent intramammary infection (IMI) with non-aureus staphylococci (NAS) within the first 4 d after calving in dairy heifers affects quarter milk yield (qMY) and quarter milk somatic cell count (qSCC) during the first 4 mo of lactation. In total, 324 quarters from 82 Holstein Friesian heifers from 3 commercial dairy herds equipped with an automatic milking system were included and followed from calving up to 4 mo in lactation. The automatic milking system allowed us to precisely determine the daily qMY. A milk sample from each quarter was collected in early lactation (between 1 and 4 d in milk) for bacteriological culturing and measurement of the qSCC. Subsequently, milk samples were taken on a biweekly basis for measurement of the qSCC. The milk prolactin level in early lactation was measured, and the relation with NAS IMI was determined. Overall, NAS IMI in early lactation caused only a slight but significant increase in qSCC compared with milk from noninfected quarters during the first 4 mo in lactation, whereas no significant difference in daily qMY was present between NAS-infected and noninfected quarters. The milk prolactin level in early lactation did not differ between NAS-infected and noninfected quarters either. Our data suggest that IMI with NAS (as a group) present shortly after calving do not have an adverse effect on later production. The milk prolactin concentrations were not dissimilar between NAS-infected and noninfected quarters and thus cannot explain why NAS-infected quarters do not produce less than noninfected quarters.     

Sources of variation in milk flow characteristics at udder and quarter levels

The aim of this study was to describe and analyze effects of parity, stage of lactation, milkability (3 groups of cows with differing peak flow rates), time of milking, and quarter position on milk production and milk flow measures at udder and quarter levels. Particular emphasis was put on changes to the decline phase and in duration of overmilking. More than 75,800 quarter milk flow curves and more than 19,300 udder milk flow curves obtained from 38 cows throughout lactation were analyzed. Stage of lactation significantly influenced all studied variables at both udder and quarter levels. At the quarter level, the duration of decline phase and the decline ratio (decline phase as a percentage of milking time) decreased from mo 1 to 2 and then gradually increased as lactation advanced. In contrast, at the udder level, duration of decline phase decreased throughout lactation but beginning at mo 2, the decline ratio increased as lactation advanced. The duration of the overmilking phase of quarters increased from mo 1 to 3 and then decreased in the course of lactation. Parity did not influence peak and average flow rates, the duration of increase phase, or the decline ratio at either udder or quarter levels. All milk flow measures were higher during morning milking except the duration of increase and decline phases at the quarter level and the duration of increase phase at the udder level. Milk yield and the duration of increase phase were not affected by milkability at either level. Quarters from udders with high milkability had longest duration of decline phase and the shortest overmilking phase. Milkability did not influence duration of the decline phase at the udder level. Quarter position influenced all measured variables of milk yield and milk flow. Rear quarters had significantly higher milk yield, longer time of milking, higher peak, and higher average flow rates than front quarters. Front quarters had shorter duration of increase and decline phases than rear quarters. The duration of the overmilking phase was almost double for front quarters. There were also differences in measured flow rates between left or right quarters on respective front or rear positions. Measured characteristics reported in this study may be important in setting default parameters in automated milking systems.     

基于双向电泳和质谱联用技术的水牛乳源酪蛋白的研究

利用双向电泳和基质辅助激光解吸电离飞行时间质谱(MALDI-MS)联用技术对水牛乳酪蛋白与其他乳源蛋白差异性进行了研究。根据Image Master 2D Platinum图像分析软件对不同乳源酪蛋白的双向电泳(2-DE)图谱进行蛋白斑点的匹配分析,获得21个存在于水牛奶中,主要分布在低丰度蛋白区的差异蛋白点,经质谱分析,得到4个属于水牛奶酪蛋白的主要组分,另外发现两个与水牛奶中的蛋白有较高同源性的新组分。     

Short communication: Correlations between udder morphology, milk yield, and milking ability with different milking frequencies in dairy goats

Tinerfeñ a breed goats were assigned to 2 experimental herds and milked once (n = 28) or twice (n = 24) daily to study correlations between udder morphology, milk yield, and milking ability during the middle stage of the first lactation. Pearson correlation coefficients were significantly higher between yield and measures of udder globulousness (udder volume, r = 0.79 and r = 0.59; perimeter of insertion of the udder, r = 0.47 and r = 0.37; distance between teats, r = 0.77 and r = 0.28, for goats milked once and twice daily, respectively) than for length parameters (cistern floor distance, r = 0.40 and r = −0.29; udder depth, r = −0.20 and r = 0.20). The globulousness of the udder was correlated with easier milking ability, as shown by milk fractioning (r = 0.49 to 0.70) and milk flow measures (r = 0.32 to 0.49). The results showed that the globulousness of the udder is more important than length measurements in assessing milk yield and milking ability.     

水牛乳中可培养乳酸菌多样性分析

采用传统分离培养方法,从三品杂交生水牛奶混合样品中,分离出105株乳酸菌,通过形态、生理生化、API细菌鉴定系统及16S rDNA基因序列分析方法对各菌株属种进行鉴定。16S rRNA序列分析结果显示,105株菌共分为5个属8个种,呈现较为丰富的乳酸菌多样性,具体数量分布为乳酸乳球菌21株,植物乳杆菌19株,格氏乳球菌17株,乳明串珠菌13株,食窦魏斯氏菌11株,肠膜明串珠菌8株,类肠膜魏斯氏菌6株,嗜热链球菌5株,糊精乳杆菌5株。由此可知,水牛乳中可培养乳酸菌优势菌群的主次关系为：乳酸乳球菌（Lactococcus lactis）＞植物乳杆菌（Lactobacillus plantaru）＞格氏乳球菌（Lactococcus garvieae）＞乳明串珠菌（Leuconostoc lactis）＞食窦魏斯氏菌（Weissella cibaria）,此为后续开发水牛乳中优势乳酸菌资源提供了良好的理论基础。     

Short communication: Validation of a novel milk progesterone-based tool to monitor luteolysis in dairy cows using cost-effective,on-farm measured data

The progesterone (P4) monitoring algorithm using synergistic control (PMASC) uses luteal dynamics to identify fertility events in dairy cows. This algorithm employs a combination of mathematical functions describing the increasing and decreasing P4 concentrations during the development and regression of the corpus luteum and a statistical control chart that allows identification of luteolysis. The mathematical model combines sigmoidal functions from which the cycle characteristics can be calculated. Both the moment at which luteolysis is detected and confirmed by PMASC, as well as the model features themselves, can be used to inform the farmer on the fertility status of the cows.     

Genome-wide association studies to identify quantitative trait loci affecting milk production traits in water buffalo

Water buffalo is the second largest resource of milk supply around the world, and it is well known for its distinctive milk quality in terms of fat, protein, lactose, vitamin, and mineral contents. Understanding the genetic architecture of milk production traits is important for future improvement by the buffalo breeding industry. The advance of genome-wide association studies (GWAS) provides an opportunity to identify potential genetic variants affecting important economical traits. In the present study, GWAS was performed for 489 buffaloes with 1,424 lactation records using the 90K Affymetrix Buffalo SNP Array (Affymetrix/Thermo Fisher Scientific, Santa Clara, CA). Collectively, 4 candidate single nucleotide polymorphisms (SNP) in 2 genomic regions were found to associate with buffalo milk production traits. One region affecting milk fat and protein percentage was located on the equivalent of Bos taurus autosome (BTA)3, spanning 43.3 to 43.8 Mb, which harbored the most likely candidate genes MFSD14A, SLC35A3, and PALMD. The other region on the equivalent of BTA14 at 66.5 to 67.0 Mb contained candidate genes RGS22 and VPS13B and influenced buffalo total milk yield, fat yield, and protein yield. Interestingly, both of the regions were reported to have quantitative trait loci affecting milk performance in dairy cattle. Furthermore, we suggest that buffaloes with the C allele at AX-85148558 and AX-85073877 loci and the G allele at AX-85106096 locus can be selected to improve milk fat yield in this buffalo-breeding program. Meanwhile, the G allele at AX-85063131 locus can be used as the favorable allele for improving milk protein percentage. Genomic prediction showed that the reliability of genomic estimated breeding values (GEBV) of 6 milk production traits ranged from 0.06 to 0.22, and the correlation between estimated breeding values and GEBV ranged from 0.23 to 0.35. These findings provide useful information to understand the genetic basis of buffalo milk properties and may play a role in accelerating buffalo breeding programs using genomic approaches.     

Short communication: Relationship of blood DNA methylation rate and milk performance in dairy cows

The objective of this study was to investigate the global methylation rate in blood DNA and its relationship with lactation performance. A total of 196 mid-lactation dairy cows were fed the same diet under the same management. Milk yield was recorded and blood samples were collected from the jugular vein before morning feeding. The blood global DNA methylation rates were quantified using a methylation quantification kit. Overall, the average blood global DNA methylation rate of all cows was 12.4%. When DNA methylation rates were compared between cows with high (n = 40; 37.0 to 42.0 kg/d) and low (n = 33; 24.0 to 30.0 kg/d) milk yield, DNA methylation rates in the lower-yield cows (14.1 ± 0.7%) were significantly higher than those in the higher-yield animals (11.6 ± 0.7%). Our results indicated an association of milk and protein yields with global DNA methylation rates in lactating dairy cows. However, further research is needed to determine whether this association reflects the true influence of epigenetic mechanisms on yield or whether other factors, such as different proportions of blood cell types in high- and low-yielding cows, affect apparent global DNA methylation levels.     

Short communication: Detection of human Torque teno virus in the milk of water buffaloes (Bubalus bubalis)

Forty-four raw milk and 15 serum samples from 44 healthy water buffaloes reared in Caserta, southern Italy, the most important region in Europe for buffalo breeding, were examined to evaluate the presence of Torque teno viruses (TTV) using molecular tools. Furthermore, 8 pooled pasteurized milk samples (from dairy factories having excellent sanitary conditions) and 6 Mozzarella cheese samples were also tested. Four of the cheese samples were commercial Mozzarella cheese; the remaining 2 were prepared with TTV-containing milk. Human TTV were detected and confirmed by sequencing in 7 samples of milk (approximately 16%). No TTV were found in serum, pooled pasteurized milk, or Mozzarella cheese samples. The samples of Mozzarella cheese prepared with TTV-containing milk did not show any presence of TTV, which provides evidence that standard methodological procedures to prepare Mozzarella cheese seem to affect viral structure, making this food fit for human consumption. The 7 TTV species from water buffaloes were identified as genotypes corresponding to the tth31 (3 cases), sle 1981, sle 2031, and NLC030 (2 cases each) human isolates. Although cross-species infection may occur, detection of TTV DNA in milk but not in serum led us to believe that its presence could be due to human contamination rather than a true infection. Finally, the mode of transmission of TTV has not been determined. Contaminated of the food chain with TTV may be a potential risk for human health, representing one of the multiple routes of infection.     

The prevalence and characterization of Bacillus cereus isolated from raw and pasteurized buffalo milk in southwestern China

Bacillus cereus is an important food-borne pathogenic bacteria and a putrid microorganism in the dairy industry. Raw and pasteurized buffalo milk play important roles in the dairy market in southwestern China. However, the reports on the prevalence and characterization of B. cereus strains isolated from the above sources are lacking. In this study, 150 raw buffalo milk samples and 300 pasteurized buffalo milk samples were collected from 3 provinces in southwestern China. The genotype, virulence gene distribution, antibiotic resistance, and biofilm-forming ability of isolates were analyzed. Ninety-six B. cereus strains were isolated and identified: 50 isolates (33.3%) from buffalo raw milk and 46 isolates (15.3%) from pasteurized buffalo milk. These strains were classified into 41 sequence types (ST) and 5 groups, of which ST857 was the predominant ST. The detection rates of virulence genes nheABC cluster, hblACD cluster, cytK, bceT, entFM, hlyII, and cesB were 89.6%, 13.5%, 64.6%, 71.9%, 84.4%, 62.5%, and 6.25%, respectively. The antimicrobial susceptibility testing showed that more than 90% of the isolates were susceptible to gentamicin, chloramphenicol, ciprofloxacin, erythromycin, vancomycin, and tetracycline, as well as resistant to ampicillin, cefepime, oxacillin, and rifampin. The results of biomass biofilm evaluation of the isolates on the stainless-steel tube showed that the optical density values at a wavelength of 595 nm of all strains in group I were greater than 1, with the strongest overall biofilm-forming ability among 5 groups, and the overall biofilm-forming ability of group III was the weakest. There was a relationship between the biofilm-forming ability and phylogenetic relationship of B. cereus strains. Taken together, our findings are the first to report the contamination situation and characterization of B. cereus isolated from raw and pasteurized buffalo milk in southwestern China as well as indicate the potential risk posed by this pathogen to dairy industry and public health.