UHT milk contains multiple forms of αS1-casein that undergo degradative changes during storage

Milk proteins are susceptible to chemical changes during processing and storage. We used proteomic tools to analyse bovine αS1-casein in UHT milk. 2-D gels of freshly processed milk αS1-casein was presented as five or more spots due to genetic polymorphism and variable phosphorylation. MS analysis after phosphopeptide enrichment allowed discrimination between phosphorylation states and genetic variants. We identified a new alternatively-spliced isoform with a deletion of exon 17, producing a new C-terminal sequence, K164SQVNSEGLHSYGL177, with a novel phosphorylation site at S174. Storage of UHT milk at elevated temperatures produced additional, more acidic αS1-casein spots on the gels and decreased the resolution of minor forms. MS analysis indicated that non-enzymatic deamidation and loss of the N-terminal dipeptide were the major contributors to the changing spot pattern. These results highlight the important role of storage temperature in the stability of milk proteins and the utility of proteomic techniques for analysis of proteins in food.

It will be succesful to have a commercial relation between France and Colombia focus on the milk sector ?

Globalization has led to a drastic change on the international trade, this has caused that many countries such as France and Colombia find new business partners. This is the raison because I develop the investigation about How these countries can be integrated commercially in the dairy sector? Colombia and France had active trade relations for more than 8 years, but this has been declining for several factors. In the case of France, it has found that Germany is an excellent producer of raw milk, which can supply domestic demand and can export large quantities with a low price to European countries because of its proximity. For this reason, Colombia is a country that can not compete with Germany directly in raw milk, but I concluded that Colombia could become a major competitor with organic milk. This is the result of a research of the way they raise and feed the cows in the two countries, Colombia is generally on open country and Germany is on cow housing. In Colombia, the country found that Venezuela could offer many benefits in the processing of raw milk, and other processes that require high technology. But today Colombia has had several disputes with Venezuela and many Colombian companies are affected in this sector. Additionally, France is one of the largest producers and distributors of processed milk and its derivatives; it launches annually many manufactured products with high technology. At this point, Colombia can take out the best advantage because it can create an strategic alliance with French companies to bring most innovative and processed products such as cheese and yoghurts. The theoretical framework of this thesis consists on the analysis of competitiveness, because it is relevant to see if these two countries are competitive or if one has the comparative advantage over the other. The related authors are: Michael Porter, Adam Smith and David Ricardo. To complete the theoretical part, we found that France has a comparative advantage over Colombia in this sector thanks to its high technology, and Colombia is not currently competitive in the area of raw milk because the milk price is higher and the distance is a barrier, and it has no technology. But this research indicates that Colombia could become a competitive country selling organic milk.

Effect of somatic cell counts on lipolysis, proteolysis and apparent viscosity of UHT milk during storage

In this work, lipolysis, proteolysis and viscosity of ultra-high temperature (UHT) milk containing different somatic cell counts (SCC) were investigated. UHT milks were analysed on days 8, 30, 60, 90 and 120 of storage. Lipolysis as measured by free fatty acids increase, casein degradation and viscosity of UHT milk were not affected by SCC but increased during storage. A negative relationship was observed between SCC and casein as a percentage of true protein on the 120th day of storage, hence indicating that high SCC increases the proteolysis of UHT milk by the end of its shelf life.

Optimization of the rheological properties of probiotic yoghurts supplemented with milk proteins

This study aimed to optimize the rheological properties of probiotic yoghurts supplemented with skimmed milk powder (SMP) whey protein concentrate (WPC) and sodium caseinate (Na-Cn) by using an experimental design type simplex-centroid for mixture modeling It Included seven batches/trials three were supplemented with each type of the dairy protein used three corresponding to the binary mixtures and one to the ternary one in order to increase protein concentration in 1 g 100 g(-1) of final product A control experiment was prepared without supplementing the milk base Processed milk bases were fermented at 42 C until pH 4 5 by using a starter culture blend that consisted of Streptococcus thermophilus Lactobacillus delbrueckii subsp bulgaricus and Bifidobacterium (Humans subsp lactis The kinetics of acidification was followed during the fermentation period as well the physico-chemical analyses enumeration of viable bacteria and theological characteristics of the yoghurts Models were adjusted to the results (kinetic responses counts of viable bacteria and theological parameters) through three regression models (linear quadratic and cubic special) applied to mixtures The results showed that the addition of milk proteins affected slightly acidification profile and counts of S thermophilus and B animal`s subsp lactis but it was significant for L delbrueckii subsp bulgaricus Partially-replacing SMP (45 g/100 g) with WPC or Na-Cn simultaneously enhanced the theological properties of probiotic yoghurts taking into account the kinetics of acidification and enumeration of viable bacteria (C) 2010 Elsevier Ltd All rights reserved

Hazards in non-pasteurized milk on retail sale in Brazil: prevalence of Salmonella spp, Listeria monocytogenes and chemical residues

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nonculture-based identification of bacteria in milk by protein fingerprinting

Traditional methods for bacterial identification include Gram staining, culturing, and biochemical assays for phenotypic characterization of the causative organism. These methods can be time-consuming because they require in vitro cultivation of the microorganisms. Recently, however, it has become possible to obtain chemical profiles for lipids, peptides, and proteins that are present in an intact organism, particularly now that new developments have been made for the efficient ionization of biomolecules. MS has therefore become the state-of-the-art technology for microorganism identification in microbiological clinical diagnosis. Here, we introduce an innovative sample preparation method for nonculture-based identification of bacteria in milk. The technique detects characteristic profiles of intact proteins (mostly ribosomal) with the recently introduced MALDI SepsityperTM Kit followed by MALDI-MS. In combination with a dedicated bioinformatics software tool for databank matching, the method allows for almost real-time and reliable genus and species identification. We demonstrate the sensitivity of this protocol by experimentally contaminating pasteurized and homogenized whole milk samples with bacterial loads of 10(3)-10(8) colony-forming units (cfu) of laboratory strains of Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus. For milk samples contaminated with a lower bacterial load (104 cfu mL-1), bacterial identification could be performed after initial incubation at 37 degrees C for 4 h. The sensitivity of the method may be influenced by the bacterial species and count, and therefore, it must be optimized for the specific application. The proposed use of protein markers for nonculture-based bacterial identification allows for high-throughput detection of pathogens present in milk samples. This method could therefore be useful in the veterinary practice and in the dairy industry, such as for the diagnosis of subclinical mastitis and for the sanitary monitoring of raw and processed milk products.

Comparison of texture of yogurt made from conventionally treated milk and UHT milk fortified with low-heat skim milk powder

The textures of yogurt made from ultra-high temperature (UHT) treated and conventionally treated milks at high total solids were investigated. The yogurt premixes, fortified with low-heat skim milk powder to 16%, 18%, and 20% total solids, were UHT processed at 143 degreesC for 6 s and heated at 85 degreesC for 30 min using the conventional method. The onset of gelation was delayed in the UHT-processed milk compared with conventionally heated milk. During fermentation, the viscosity of yogurt made, from UHT-treated milk at 20% total solids was close to that of yogurt made from conventionally treated milk with 16% total solids. However, after storage for greater than or equal to1 d, the yogurt made from UHT-treated milk had lower viscosity and gel strength than the yogurt made from conventionally treated milk. The solids level had no influence on yogurt culture growth.

Solid phase microextraction of stale flavour volatiles from the headspace of UHT milk

Solid phase microextraction (SPME) offers a solvent-free and less labour-intensive alternative to traditional flavour isolation techniques. In this instance, SPME was optimised for the extraction of 17 stale flavour volatiles (C3-11,13 methyl ketones and C4-10 saturated aldehydes) from the headspace of full-cream ultrahigh-temperature (UHT)-processed milk. A comparison of relative extraction efficiencies was made using three fibre coatings, three extraction times and three extraction temperatures. Linearity of calibration curves, limits of detection and repeatability (coefficients of variation) were also used in determining the optimum extraction conditions. A 2 cm fibre coating of 50130 gm divinylbenzene/Carboxen/polydimethylsiloxane in conjunction with a 15 min extraction at 40 degrees C were chosen as the final optimum conditions. This method can be used as an objective tool for monitoring the flavour quality of UHT milk during storage. (c) 2005 Society of Chemical Industry.

Stale flavour volatiles in Australian commercial UHT milk during storage

Methyl ketones, aldehydes and free saturated fatty acids were measured in the headspace of samples of two indirectly processed and two directly processed Australian commercial UHT milks during room temperature storage for 16 weeks. The analytes were isolated using headspace solid phase microextraction and analysed by gas chromatography coupled with flame ionisation detection. All methyl ketones and aldehydes increased during storage, With free saturated fatty acids exhibiting little change. On average, the total methyl ketone and aldehyde concentrations in the indirectly processed UHT milks were higher than those in the directly processed samples. A strong correlation was found between the concentration of methyl ketones and various heat indices (furosine, lactulose and undenatured whey proteins) in the milk samples.

Survival of probiotics encapsulated in chitosan-coated alginate beads in yoghurt from UHT- and conventionally treated milk during storage

Survival of the microencapsulated probiotics, Lactobacillus acidophilus 547, Bifidobacterium bifidum ATCC 1994, and Lactobacillus casei 01, in stirred yoghurt from UHT- and conventionally treated milk during low temperature storage was investigated. The probiotic cells both as free cells and microencapsulated cells (in alginate beads coated with chitosan) were added into 20 g/100 g total solids stirred yoghurt from UHT-treated milk and 16 g/100 g total solids yoghurt from conventionally treated milk after 3.5 h of fermentation. The products were kept at 4 degrees C for 4 weeks. The survival of encapsulated probiotic bacteria was higher than free cells by approximately 1 log cycle. The number of probiotic bacteria was maintained above the recommended therapeutic minimum (10(7) cfu g(-1)) throughout the storage except for R bifidum. The viabilities of probiotic bacteria in yoghurts from both UHT- and conventionally treated milks were not significantly (P > 0.05) different. (c) 2004 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

An approach to identifying factors affecting milk protein concentration in dairy cattle

Milk protein production can be influenced by several factors, including environment, disease status, parity, stage of lactation, breed, genetic merit and the nutritional status of the animal (DePeters and Cant 1992). A combination of, or an interaction between, these factors can significantly affect milk protein production. Our study aims to identify the main factors affecting milk protein concentration in dairy cattle in the south-east Queensland region.

Avaliação, identificação e atividade enzimática de bactérias psicotróficas presentes no leite cru refrigerado

Extended storage of refrigerated milk can lead to reduced quality of raw and processed milk, which is a consequence of the growth and metabolic activities of psychrotrophic bacteria, able to grow under 7oC or lower temperatures. Although most of these microorganisms are destroyed by heat treatment, some have the potential to produce termoresistant proteolytic and lipolytic enzymes that can survive even UHT processing and reduce the processed products quality. Recently, the IN 51 determineds that milk should be refrigerated and stored at the farm what increased the importance of this group of microorganisms. In this work, psychrotrophic bacteria were isolated from 20 communitarian bulk tanks and 23 individual bulk tanks from dairy farms located at Zona da Mata region of Minas Gerais State and from southeastern Rio de Janeiro. Selected milk dilutions were plated on standard agar and after incubation for 10 days at 7oC, five colonies were isolated, firstly using nutrient agar and after using McConkey agar for 24 hours at 21oC. The isolates were identified by morphology, Gram stain method, catalase production, fermentative/oxidative metabolism and by API 20E, API 20NE, API Staph, API Coryne or API 50 CH (BioMerieux). In order to ensure reproductibility, API was repeated for 50% of the isolates. Species identification was considered when APILAB indexes reached 75% or higher. 309 strains were isolated, 250 Gram negative and 59 Gram positive. 250 Gram negative isolates were identified as: Acinetobacter spp. (39), Aeromonas spp. (07), A. Hydrophila (16), A. sobria (1), A. caviae (1), Alcaligenes feacalis (1), Burkholderia cepacia (12), Chryseomonas luteola (3), Enterobacter sp. (1), Ewingella americana(6), Hafnia alvei (7), Klebsiella sp. (1), Klebsiella oxytoca (10), Yersinia spp. (2), Methylobacterium mesophilicum (1), Moraxella spp. (4), Pantoea spp. (16), Pasteurella sp. (1), Pseudomonas spp. (10), P. fluorescens (94), P. putida (3), Serratia spp. (3), Sphigomonas paucomobilis (1). Five isolates kept unidentified. Pseudomonas was the predominant bacteria found (43%) and P. fluorescens the predominant species (37.6%), in accordance with previous reports. Qualitative analysis of proteolytic and lipolytic activity was based on halo formation using caseinate agar and tributirina agar during 72 hours at 21oC and during 10 days at 4°C, 10oC and 7°C. Among 250 Gram negative bacteria found, 104 were identified as Pseudomonas spp. and 60,57% of this group showed proteolytic and lipolytic acitivities over all four studied temperatures. 20% of Acinetobacter, Aeromonas, Alcaligenes, Burkholderia, Chryseomonas, Methylobacterium, Moraxella presented only lipolytic activity. Some isolates presented enzymatic activity in one or more studied temperatures. Among Gram positive bacteria, 30.51% were proteolytic and lipolytic at 10oC, 8.47% were proteolytic at 7oC, 10oC, and 21oC, 8.47% were proteolytic at all studied temperatures (4oC, 7oC, 10oC and 21oC) and 3.38% were proteolytic only at 21oC. At 4oC, only one isolate showed proteolytic activity and six isolates were lipolytic. In relation to Gram negative microorganisms, 4% were proteolytic and lipolytic at 7oC, 10oC and 21oC, 10% were proteolytic at 10oC and 4.4% were lipolytic at 4oC, 7oC, 10oC and 21oC, while 6.4% of all isolates were proteolytic and lipolytic at 10oC and 21oC as well as lipolytic at 4oC and 7oC. These findings are in accordance with previous researches that pointed out Pseudomonas as the predominant psycrotrophic flora in stored refrigerated raw milk

Effect of high hydrostatic pressure processing on rheological and textural properties of probiotic low-fat yogurt fermented by different starter cultures

The effect of milk processing on rheological and textural properties of probiotic low-fat yogurt (fermented by two different starter cultures) was studied. Skim milk fortified with skim milk powder was subjected to three treatments: (1) thermal treatment at 85C for 30 min; (2) high hydrostatic pressure (HHP) at 676 MPa for 5 min; and (3) combined treatments of HHP (676 MPa for 5 min) and heat (85C for 30 min). The processed milk was fermented using two different starter cultures containing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus and Bifidobacterium longum at inoculation rates of 0.1 and 0.2%. Rheological parameters were determined and a texture profile analysis was carried out. Yogurts presented different rheological behaviors according to the treatment used, which could be attributed to structural phenomena. The combined HHP and heat treatment of milks resulted in yogurt gels with higher consistency index values than gels obtained from thermally treated milk. The type of starter culture and inoculation rate, providing different fermentation pathways, also affected the consistency index and textural properties significantly. The combined HHP and heat treatment of milks before fermentation, and an inoculation rate of 0.1% (for both cultures), led to desirable rheological and textural properties in yogurt, which presented a creamy and thick consistency that does not require the addition of stabilizers.

High hydrostatic pressure processing on microstructure of probiotic low-fat yogurt

The effect of milk processing on the microstructure of probiotic low-fat yogurt was studied. Skim milk fortified with skim milk powder was subjected to three treatments prior to innoculation: thermal treatment at 85 degrees C for 30 min, high hydrostatic pressure at 676 MPa for 5 min, and combined treatments of high hydrostatic pressure (HHP) and heat. The processed milk was then fermented by using two different starter cultures containing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Bifidobacterium longum. The microstructure of heat-treated milk yogurt had fewer interconnected chains of irregularly shaped casein micelles, forming a network that enclosed the void spaces. on the other hand, microstructure of HHP yogurt had more interconnected clusters of densely aggregated protein of reduced particle size, with an appearance more spherical in shape, exhibiting a smoother more regular surface and presenting more uniform size distribution. The combined HHP and heat milk treatments led to compact yogurt gels with increasingly larger casein micelle clusters interspaced by void spaces, and exhibited a high degree of cross-linking. The rounded micelles tended to fuse and form small irregular aggregates in association with clumps of dense amorphous material, which resulted in improved gel texture and viscosity. (C) 2007 Elsevier Ltd. All rights reserved.

Determinação de GMP e CMP* no leite por métodos espectrofotométrico (ANSM) e cromatográfico (HPLC) - parâmetros metodológicos (*glicomacropeptídeo e caseinomacropeptídeo)

Glycomacropeptide is a glycosilated fraction of bovine kappa-casein that remains soluble when milk is clotted by rennin. Determinations of milk sialic acid content are useful because its concentration reflects the amount of free GMP of milk. In normal milk these amounts are very low, 12 to 16 times lower than in sweet whey. Therefore, its determination may be applied to verify possible frauds with whey addictions, since it works as a fingerprint. With the description of a new spectrophotometric method for determination of free GMP (ANSM) occurred a simplification of procedures, being faster than others (HPLC method), without loss of accuracy. However, due to variations of glycosilation in kappa-casein between animals, during the lactation period, due to mastitis and yet due to proteolysis on milk, it was necessary to know these variations to interpret correctly the analytical results. It was analyzed 1,703 samples of producer's raw milk and 1,189 samples of processed milk (HTST and UHT). The results showed that normal milk from herd (producer's milk) have only small amounts of free GMP, with A470nm = 0.232±0.088 or 3.89±1.25 mg of sialic acid/L. The upper limit of this distribution was A = 0.496; thus every bigger value may represent a problem, being outside of normal distribution.