Effects of somatic cell counts in milk on physical and chemical characteristics of yoghurt

The quality of plain stirred yoghurt produced from whole milk with somatic cell counts (SCC) at low (147,000 cells mL⁻¹), intermediate (434,000 cells mL⁻¹) and high (1,943,000 cells mL⁻¹) levels was examined. Each milk treatment was obtained from selected cows, according to its SCC status and milk composition. Yoghurt samples were analysed on days 1, 10, 20 and 30 after production. Analyses included pH, acidity, fat, lipolysis (expressed as free fatty acids, FFA), proteolysis and apparent viscosity. Viscosity of high SCC yoghurt was higher (P<0.05) than the low SCC yoghurt on days 10, 20 and 30 of storage. High SCC yoghurt also had higher FFA content (P<0.05). SCC did not affect pH, acidity, fat content and proteolysis of the yoghurt (P>0.05). Results indicate that SCC in milk increases the lipolysis in the resulting yoghurt during storage for 30 d.     

Milk with different somatic cells counts and the physicochemical,microbiological characteristics and fatty acid profile of pasteurised milk cream: is there an association?

In this study, raw cow milk containing somatic cells counts (SCC) at mean levels of 39 000 cells mL^?1 (low), 349 000 cells mL^?1 (intermediate) and 1 297 000 cells mL^?1 (high) was used for the production of pasteurised cream. Physicochemical (pH, fat and fatty acid profile) and microbiological analyses (mesophilic and psychrotrophs) were performed in the obtained creams during 30 days of refrigerated storage at 5 °C ± 2. No interactions (P > 0.05) were found between SCC, storage time and the physicochemical and microbiological variables studied. Fatty acid profile was similar among the SCC creams, except for oleic acid (C18:1), which decreased (P < 0.05) in intermediate and high SCC creams. Considering the technological aspect, our findings suggest that milk cream manufacture can be an interesting option for the use of high SCC milk.     

Somatic cell counts,chemical composition and coagulation properties of goat and sheep bulk tank milk

This study focused on the need for bulk milk tank somatic cell count (BMTSCC) thresholds and cut-off levels indicating a decrease in milk quality that consequently influences product quantity and quality. First, 226 ewes and 231 goat bulk tank milk samples were collected from different Israeli herds and coagulation properties were determined. Second, soft cheese was produced. No correlation of coagulation properties was found with BMTSCC for sheep milk up to 3264 × 10³ and goat milk up to 6452 × 10³ cells mL⁻¹. Coagulation properties of goat milk with cell count higher than the latter resulted in a significant decrease in curd firmness. For breeds and management system in Israel, 2500 × 10³ cells mL⁻¹ is suggested as the cut-off level for sheep and 3500 × 10³ cell mL⁻¹ for goats. The cell count cut-off level and milk price according to BMTSCC should be tested and then determined for every breed and management and final dairy product.     

Chemical composition,protein fraction and fatty acid profile of donkey milk during lactation

The chemical composition, fatty acid and protein fraction of donkey milk samples from domestic (Arcadian) breed were assessed from the 30th day to the 210th day of lactation. Total fat, proteins and minerals (Ca, Mg, K and P) were affected by the stage of lactation. Fatty acids were significantly influenced, with a decrease of 31.4% in saturated fatty acids, an increase of 53.8% in unsaturated fatty acids, a concomitant elevation of polyunsaturated fatty acids and a low n-6/n-3 ratio (2:1). Analysis by sodium dodecylsulphate-polyacrylamide gel electrophoresis found, on average, 42.8% casein and 57.2% whey protein. The concentration of lysozyme was between 1.20 and 2.54 mg mL⁻¹, while the highest values were detected by the lysoplate method. Finally, low microbial content and somatic cells were also found in donkey milk with overall average log 4.4 cfu mL⁻¹ and log 4.8 cells mL⁻¹, respectively.     

HPLC of proteose peptones for evaluating ageing of packaged pasteurized milk

The proteolysis of casein (CN) occurring in packaged pasteurized milk (PM) during refrigerated storage was studied with relation to hygienic and microbiological characteristics of starting raw milk. Six batches of raw milk having standard plate count (SPC) from 1.5×10⁴ to 2.5×10⁵ cfu mL⁻¹ and somatic cell count (SCC) from 1.6×10⁵ to 4.4×10⁵ units mL⁻¹ were pasteurized (73 °C for 15 s), packaged and stored at 4 °C for 12 days. Capillary zone electrophoresis of CN showed breakdown of β-CN in all PM samples during storage. An HPLC method for monitoring proteose peptones (PP) formation was developed. Level of PP in PM samples increased, with keeping time from 667–789 to 947–1383 mg L⁻¹ and PP formation was significantly (P<0.05) related to SCC of starting raw milk. Electrospray ionization–mass spectrometry showed that PP were mainly represented by PP-5 from either A¹ or A² variants of β-CN. Five commercial samples of PM were analysed for PP formation during 14-day storage at 4 °C. Commercial samples prepared by microfiltration process or bactofugation combined with pasteurization showed the slowest formation of PP. The effect of storage temperature on PP formation was evaluated by keeping a conventional PM sample at either 8 or 12 °C for 12 days. Proteolysis of all major CNs upon action of plasmin and bacterial proteinases was observed under these conditions. PP level thus proves to be a reliable analytical index for evaluating the ageing of packaged PM during refrigerated storage.     

Somatic cell recovery by microfiltration technologies: A novel strategy to study the actual impact of somatic cells on cheese matrix

The actual impact of the somatic cells in the dairy technology is still ill-defined, because the increase in milk somatic cell count, usually correlated with mastitis factors, impairs the raw milk composition, through mainly unwanted proteolysis and lipolysis. This study used microfiltration technologies for recovering high quantity of somatic cells and for clarifying their role in cheese quality. Three series of Swiss-type cheeses were manufactured by adding 0 (control), 4 × 10⁵ and 9 × 10⁵ somatic cells mL⁻¹. These cells were traced for the first time during the cheese making process by using adapted flow cytometry and real-time quantitative PCR. Proteolysis and lipolysis indices were measured throughout ripening time. Only a weak increase in lipolysis (+28%) and proteolysis (+8%) was observed in the highest somatic cell count cheese, despite 73% of the cells trapped within the cheeses. Our approach gives a new view of somatic cell role in cheese milk alteration.     

Physicochemical traits and sensory quality of commercial butter produced in the Azores

In the volcanic islands of the Azores archipelago, pasture is available the entire year and affects the composition of bovine milk fat. The composition of cows' milk butter manufactured in this region was evaluated. All brands showed excellent microbiological and oxidative quality. The fatty acid profile range presented a low n-6/n-3 ratio (1.81–3.38), low atherogenic (2.86–3.11) and thrombogenicity (3.27–3.60) indices and reduced cholesterol content (136–143 mg 100 g⁻¹). In addition, Azorean butters have a high content of conjugated linoleic acid (0.44–0.64 mg 100 g⁻¹), β-carotene (0.12–0.17 mg 100 g⁻¹) and α-tocopherol (1.40–2.20 mg 100 g⁻¹). Sensory analysis revealed high scores for appearance, consistency and flavour of all brands. These results indicate that Azorean butters produced from cows’ milk based on grass-feeding have a potentially healthier fat content and a desirable flavour that, associated with its “natural image”, may be promoted for product differentiation.     

Impact of β-glucan on debittering,bioaccessibility and storage stability of skim milk fortified with shrimp oil nanoliposomes

Shrimp oil was encapsulated in nanoliposomes and fortified into skim milk. Shrimp oil nanoliposomes (SONL) were thermodynamically stable when added into skim milk at 10 mL 100 mL⁻¹. Mild bitterness in fortified skim milk caused by the SONL was masked by adding β-glucan at various levels (0.05–0.2 g 100 mL⁻¹). With the addition of SONL, fortified skim milk appeared more reddish in colour due to the presence of astaxanthin. Addition of β-glucan resulted in the increase in viscosity of the fortified milk by forming network of junction zones. During the storage of skim milk fortified with SONL and 0.1 g 100 mL⁻¹ β-glucan at 4 °C for 15 days, no major quality changes took place. Simulated in vitro digestion studies revealed that 45.41 g 100 g⁻¹ eicosapentaenoic acid (EPA) and 48.86 g 100 g⁻¹ docosahexaenoic acid (DHA) from shrimp oil were bioaccessible for absorption in the gut after digestion.     

Effects of somatic cell counts on the physicochemical and rheological properties of yoghurt made from sheep’s milk

In the present work, yoghurts were made from sheep’s milk with two different somatic cell count (SCC), at low (200 000 cells mL^?1) and high (750 000 cells mL^?1) levels. The characteristics of the final product were analysed for pH, acidity, protein, total solids, fat, syneresis, water holding capacity (WHC) and apparent viscosity. Samples were analysed on days 1, 7 and 14 after production of yoghurts. The SCC had no significant effect either on the acidity or pH of the yoghurt at 24 h (P > 0.05) but a significant effect (P < 0.05) was observed at 168 h. No effects of SCC were observed on total solids and fat content of the yoghurt after 24 and 168 h. High SCC (HSCC) yoghurt had higher protein content (P < 0.05). The yoghurt with the highest SCC had the highest level of syneresis. Viscosity of HSCC yoghurt was higher than that of the low SCC yoghurt on days 1, 7 and 14 of storage. The flow properties also showed that the low SCC yoghurt was softer than that from milk with high content in somatic cells.     

Validation of the sensitive and accurate quantitation of the fatty acid distribution in bovine milk

A method for the precise analysis of the complex mixture of fatty acids in milk has been developed and validated. The triacylglycerol of nonanoic acid was applied as the internal standard (ISTD) for absolute quantification. Milk lipids were extracted by miniaturised ultrasonication and methylated with trimethylsulfonium hydroxide. Resulting fatty acid methyl esters were determined by gas chromatography with flame ionisation detection giving excellent resolution, including separation of several 18:1 isomers. The low quantitation limit (0.01 mg mL⁻¹ milk) indicates that the sensitivity of the method is sufficient to quantify up to 50 fatty acids, from 4:0 to 23:0. Measurements of precision provided excellent results for different bovine milk samples of different fat content (coefficient of variance: 1.9% and 9.8% for intra- and interday precision, respectively). Recovery averaged 108 ± 3.5%. Evaluation of methods for determining the total fat content showed that gravimetry is no longer needed when using the ISTD.     

Quantification of major camel milk proteins by capillary electrophoresis

Proteins from dromedary camel milk (CM) produced in Europe were separated and quantified by capillary electrophoresis (CE). CE analysis showed that camel milk lacks β-lactoglobulin and consists of high concentration of α-lactalbumin (2.01 ± 0.02 mg mL⁻¹), lactoferrin (1.74 ± 0.06 mg mL⁻¹) and serum albumin (0.46 ± 0.01 mg mL⁻¹). Among caseins, the concentration of β-casein (12.78 ± 0.92 mg mL⁻¹) was found the highest followed by α-casein (2.89 ± 0.29 mg mL⁻¹) while κ-casein represented only minor amount (1.67 ± 0.01 mg mL⁻¹). These results were in agreement with sodium dodecyl sulphate-polyacrylamide gel electrophoresis patterns. Overall, CE offers a quick and reliable method for the determination of major CM proteins, which may be responsible for the many nutritional and health properties of CM.     

Addition of milk fat globule membrane as an ingredient of infant formulas for resembling the polar lipids of human milk

Polar lipid (PL) contents in human milk (HM) from two different geographic zones in Spain (central and coastal) were determined. These PLs were also analysed in several infant formulas (IFs), three of which contained milk fat globule membrane (MFGM), an ingredient used to resemble the PL profile of HM. Total PL in HM decreased significantly (p < 0.05) from transitional milk (48.62 mg 100 mL⁻¹) to 6 months (28.66 mg 100 mL⁻¹). In HM, sphingomyelin was the most abundant PL, followed by phosphatidylethanolamine; in IFs the most abundant PL was phosphatidylethanolamine. Only IFs with MFGM (54.79–58.07 mg 100 mL⁻¹) could supply the total and individual PL content present in all lactation periods, with the exception of sphingomyelin, which did not match the content in transitional milk. PL intake by infants fed HM or IFs was determined to be 96–306 and 152–575 mg day⁻¹, respectively.     

Immunomodulatory activities of Lactobacillus rhamnosus ZDY114 and donkey milk in BALB/c mice

The immunomodulatory activity of Lactobacillus rhamnosus ZDY114 and donkey milk in BALB/c mice was evaluated by assessing the splenic lymphocyte transformation, haemolytic complement activity, carbon clearance ability and natural killer cell activity. Results showed donkey milk (5 g kg⁻¹) in combination with L. rhamnosus ZDY114 (5 × 10⁷ cfu mL⁻¹) exhibited a significant increase in splenic lymphocyte proliferation, carbon granule engulfing ability and natural killer cell activity when compared with donkey milk or L. rhamnosus ZDY114 alone (p < 0.05). An elevated response in serum haemolytic activity was only observed when compared with L. rhamnosus ZDY114. In conclusion, donkey milk (5 g kg⁻¹) in combination with L. rhamnosus ZDY114 (5 × 10⁷ cfu mL⁻¹) was able to enhance specific immune functions.     

Volatile organic compound profiles of yoghurt produced from cow's milk with different somatic cell counts

The present study aimed to investigate the effect of somatic cell count (SCC) of raw cow's milk on the volatile organic compound (VOC) profiles of yoghurt. Test yoghurt samples were produced from three batches of cow's milk with low, medium and high SCC, respectively. The VOCs were determined by gas chromatography–mass spectrometry (GC–MS) analysis. A lower diacetyl and acetoin content and a higher content of 2‐heptanone, and butanoic and hexanoic acids were established in the yoghurt samples from batches with high SCC of above 1 000 000 cells/cm³. The increased SCC of cow's milk had a negative effect on the volatile organic compound profiles of yoghurt.     

Contribution of proteolytic activity associated with somatic cells in milk to cheese ripening

The effect of proteolytic enzymes from somatic cells on cheese quality was studied. In preliminary experiments, milk and two sodium caseinate systems (pH 6.5 and pH 5.2, the latter in the presence of 5% NaCl) were used as substrates to investigate the proteolytic activity of somatic cells recovered from mastitic milk. Urea-polyacrylamide gel electrophoretograms of hydrolysates suggested that somatic cell extracts contributed directly to proteolysis both in buffer and in milk, but that such activity was reduced by batch pasteurisation (63 °C for 30 min). Sodium caseinate was readily hydrolysed by somatic cell extracts; hydrolysis of α_s1-casein was greater at pH 5.2 and increased with level of somatic cells, suggesting that somatic cells contain proteolytic enzymes which are more active at acidic pH values. Subsequently, miniature Cheddar-type cheeses were made from batches of milk to which somatic cells were added (at levels of levels of 3×10⁵ or 6×10⁵ cells mL⁻¹), either before or after pasteurisation. Proteolysis during ripening of cheese (as measured by levels of pH 4.6-soluble nitrogen) increased with somatic cell addition, although this effect was reduced by pasteurisation after cell addition. Somatic cells may also have directly influenced cheese moisture content, which has been established as a principal indicator of quality of Cheddar-type cheese. Proteolytic enzymes of somatic cells from milk were shown to contribute directly to proteolysis in milk and cheese.     

Antihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk

A total of 231 microorganisms were isolated from raw cow milk samples and the angiotensin-converting enzyme-inhibitory (ACEI) activity of the resultant fermented milk produced with the isolated microorganisms was assayed. Forty-six of these microorganisms were selected on the basis of high ACEI activity. Four Enterococcus faecalis strains stood out as producers of fermented milk with potent ACEI activity (IC₅₀ (the protein concentration that inhibits 50% of ACE activity): 34–59 μg mL⁻¹). Single doses (5 mL kg⁻¹) of the whey fraction obtained from these fermented milk samples were administered to spontaneously hypertensive rats (SHR) and to normotensive Wistar-Kyoto (WKY) rats in order to investigate their possible antihypertensive activity. Highly significant decreases in the systolic blood pressure (SBP) and in the diastolic blood pressure (DBP) were observed when the fermented milk was administered to SHR. Nevertheless, the fermented milk did not modify the SBP and the DBP of the WKY rats. Raw cow milk is an excellent source of wild lactic acid bacteria able to produce fermented milk with antihypertensive activity and antihypertensive activity of milk fermented by Enterococcus faecalis strains was associated with peptides different from Ile-Pro-Pro and Val-Pro-Pro.     

Development of an immunomagnetic separation–propidium monoazide–polymerase chain reaction assay with internal amplification control for rapid and sensitive detection of viable Escherichia coli O157:H7 in milk

A rapid, reproducible, sensitive and specific combination assay was developed for Escherichia coli O157:H7 comprising sequential immunomagnetic separation (IMS), followed by propidium monoazide (PMA), exclusion by viable cells, internal amplification control (IAC) and polymerase chain reaction (PCR). IMS was used to improve sensitivity, reduce detection time and eliminate false positives. PMA was used to detect viable cells, and IAC was incorporated into the system to eliminate the inhibitors of PCR from the food matrix. In the presence of inhibitors, IAC produced no signal, thereby eliminating false negative results. The results indicated that the IMS cell capture efficiency was about 90% at a concentration of 1 × 10⁶ cfu mL⁻¹ with a detection limit of 2.1 × 10² cfu mL⁻¹ for pure culture. In an unoptimised system, IMS–PMA–PCR with IAC showed a detection limit of 5 × 10³ cfu mL⁻¹ in spiked milk over a 240 min assay, faster than conventional methods of detection.     

Chemiluminescence imaging immunoassay for multiple aminoglycoside antibiotics in cow milk

A simple and rapid multiplexed direct competitive chemiluminescent (CL) imaging immunoassay had been developed for the simultaneous detection of kanamycin (KAN) and streptomycin (STR), which were widely used against bacterial infections in animals. To achieve the multiplexed detection of the two targets, a microarray format based on nitrocellulose membranes (NCMs) has been designed. Then, the peroxidase activity of the captured HRP-labelled immunocomplex in each channel was measured by an enhanced luminol-based chemiluminescent solution using a cooled low-light CCD with high resolution. For qualitative analysis, the limit of detection (LOD) was 0.5 ng mL⁻¹ for KAN and 3.13 ng mL⁻¹ for STR by the naked eye. Through the CL data collected, KAN and STR could be detected quantitatively at the LOD of 0.03 and 0.33 ng mL⁻¹, respectively. This method can be utilised as a screening test to evaluate the presence of antibiotics in milk samples.     

Fate of enterotoxigenic Staphylococcus aureus in the presence of nisin-producing Lactococcus lactis strain during manufacture of Jben,a Moroccan traditional fresh cheese

The inhibitory effect of nisin-producing Lactococcus lactis subsp. lactis UL730 on the growth of enterotoxigenic Staphylococcus aureus J10 during manufacture of Jben, a Moroccan traditional fresh cheese prepared from recombined milk, was investigated.With an inoculum level of 10³ cfu mL⁻¹, S. aureus was absent in Jben four days after inoculation when the nisin-producing lactococcus was used as lactic starter. In contrast, it survived after that period, when the starter was non-nisin-producing. No staphylococcal thermonuclease was detected in all Jben samples made from milk inoculated with S. aureus at the level of 10³ cfu mL⁻¹.With a higher inoculum of 10⁵ cfu mL⁻¹, S. aureus was still present in Jben after manufacture and persisted during the storage of the product for 3 days in the laboratory, even when the starter used was nisin-producing. Staphylococcal thermonuclease and type C enterotoxin were detected in all Jben samples made from milk inoculated with 10⁵ cfu mL⁻¹. Thermonuclease and enterotoxin were already produced in the coagulum, at 24 h after milk inoculation with S. aureus.     

1H NMR and Raman spectroscopy of oils and extracts obtained from organic and conventional goji berries: yield,fatty acids,carotenoids and biological activities

The yields of extraction, fatty acid and carotenoid compositions, antioxidant and antimicrobial activities of organic and conventional Lycium barbarum oils and extracts were investigated. Different methods were used in the extraction: Soxhlet, maceration and Bligh & Dyer. Sample characterization was performed by proton nuclear magnetic resonance and Raman spectroscopy. Soxhlet presented a higher yield for organic (4.58%) and conventional (2.83%) fruit. The Goji samples showed a high content of unsaturated fatty acids (UFA) for organic (78.29–84.72%) and conventional (70.74–80.20%) fruit. The main phytochemicals identified in the samples were linoleic acid and zeaxanthin dipalmitate. The maceration method was the most efficient in the extraction of carotenoids with high antioxidant (2.23–40.94 mmolTE 100 g⁻¹) and antimicrobial (3.12–200 mg mL⁻¹) activities. The organic goji samples showed higher yield, UFA, carotenoids and biological activities and these results indicate that this fruit is suitable for cosmetic, pharmaceutical and food applications.