RESUMO
Milk that does not coagulate after rennet addition, also called noncoagulating (NC) milk, is unwanted in cheese production due to prolonged processing time. Amounts of whey and casein proteins, genetic variants, as well as posttranslational modifications (PTM) of proteins are all contributing factors in rennet-induced coagulation of milk. In this study, we conducted a wide-ranging investigation of milk proteins in milk samples from 616 Swedish Red dairy cattle using liquid chromatography-high resolution mass spectrometry. Relative concentration of proteins, genetic variants, and PTM were compared between NC milk and coagulating milk. The PTM investigated were phosphorylation of caseins and glycosylation of κ-casein. Several genetic variants and PTM were found, including rare phosphorylation variants of the αS-caseins. Genetic variants were found to effect the expressed amount of different proteins. Further, the effect of protein amounts and PTM on a binary NC milk trait was modeled using a generalized linear model. The model showed that NC milk significantly correlated with higher relative concentrations of α-lactalbumin and ß-casein and lower relative concentrations of ß-lactoglobulin and κ-casein. Regarding PTM of caseins, an effect on NC milk from a lower relative concentration of αS1-casein with 8 phosphate groups were found, even though an effect from total relative concentration of αS1-casein was not found. This study has provided insights into protein variants and PTM important for NC milk to improve this undesirable property.
Assuntos
Proteínas do Leite/metabolismo , Leite/química , Processamento de Proteína Pós-Traducional , Animais , Caseínas/química , Bovinos , Cromatografia Líquida , Quimosina/química , Feminino , Genótipo , Lactalbumina/metabolismo , Lactoglobulinas/metabolismo , Espectrometria de Massas , Fosforilação , SuéciaRESUMO
The rennet-induced coagulation ability of milk is important in cheese production. For Swedish Red Dairy Cattle (RDC), this ability is reduced because of a high prevalence of noncoagulating (NC) milk. In this study, we simultaneously combined genetic parameters for NC milk, milk coagulation properties, milk composition, physical traits, and milk protein composition. Our aim was to estimate heritability and genetic and phenotypic correlations for NC milk and 24 traits (milk coagulation properties, milk composition, physical traits, and milk protein composition). Phenotypes and â¼7,000 SNP genotypes were available for all 600 Swedish RDC. The genotypes were imputed from â¼7,000 SNP to 50,000 SNP. Variance components and genetic parameters were estimated with an animal model. In Swedish RDC, a moderate heritability estimate of 0.28 was found for NC milk. For the other 24 traits, heritability estimates ranged from 0.12 to 0.77 (standard errors from 0.08 to 0.18). A total of 300 phenotypic and genetic correlations were estimated. For phenotypic and genetic correlations, 172 and 95 were significant, respectively. In general, most traits showing significant genetic correlations also showed significant phenotypic correlations. In this study, phenotypic and genetic correlations with NC milk suggest that many correlations between traits exist, making it difficult to predict the real consequences on the composition of milk, if selective breeding is applied on NC milk. We speculate that some of these consequences may lead to changes in the composition of milk, most likely affecting its physical and organoleptic properties. However, our results suggest that κ-casein could be used as an indicator trait to predict the occurrence of NC milk at the herd level.
Assuntos
Bovinos/genética , Quimosina/genética , Proteínas do Leite/química , Leite/química , Animais , Caseínas/química , Caseínas/genética , Bovinos/fisiologia , Queijo , Quimosina/química , Feminino , Genótipo , Proteínas do Leite/genética , Fenótipo , SuéciaRESUMO
The production of fermented milk products has increased worldwide during the last decade and is expected to continue to increase during the coming decade. The quality of these products may be optimized through breeding practices; however, the relations between cow genetics and technological properties of acid milk gels are not fully known. Therefore, the aim of this study was to identify chromosomal regions affecting acid-induced coagulation properties and possible candidate genes. Skim milk samples from 377 Swedish Red cows were rheologically analyzed for acid-induced coagulation properties using low-amplitude oscillation measurements. The resulting traits, including gel strength, coagulation time, and yield stress, were used to conduct a genome-wide association study. Single nucleotide polymorphisms (SNP) were identified using the BovineHD SNPChip (Illumina Inc., San Diego, CA), resulting in almost 621,000 segregating markers. The genome was scanned for putative quantitative trait loci (QTL) regions, haplotypes based on highly associated SNP were inferred, and the additive genetic effects of haplotypes within each QTL region were analyzed using mixed models. A total of 8 genomic regions were identified, with large effects of the significant haplotype explaining between 4.8 and 9.8% of the phenotypic variance of the studied traits. One major QTL was identified to overlap between gel strength and yield stress, the QTL identified with the most significant SNP closest to the gene coding for κ-casein (CSN3). In addition, a chromosome-wide significant region affecting yield stress on BTA 11 was identified to be colocated with PAEP, coding for ß-lactoglobulin. Furthermore, the coagulation properties of the genetic variants within the 2 genes were compared with the coagulation properties identified by the patterns of the haplotypes within the regions, and it was discovered that the haplotypes were more diverse and in one case slightly better at explaining the phenotypic variance. Besides these significant QTL comprising the 2 milk proteins, 3 additional genes are proposed as possible candidates, namely RAB22A, CDH13, and STAT1, and all have previously been found to be expressed in the mammary gland. To our knowledge, this is the first attempt to map QTL regions for acid-induced coagulation properties.
Assuntos
Bovinos/genética , Géis/química , Leite/química , Reologia , Animais , Cruzamento , Caseínas/genética , Mapeamento Cromossômico/veterinária , Produtos Fermentados do Leite/química , Feminino , Variação Genética , Estudo de Associação Genômica Ampla/veterinária , Genótipo , Haplótipos/genética , Lactoglobulinas/genética , Proteínas do Leite/metabolismo , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Locos de Características Quantitativas/genética , ViscosidadeRESUMO
Optimizing cheese yield and quality is of central importance to cheese manufacturing. The yield is associated with the time it takes before the gel has an optimal consistency for further processing, and it is well known that gel formation differs between individual milk samples. By identifying genomic regions affecting traits related to rennet-induced gelation, the aim of this study was to identify potential candidate genes affecting these traits. Hence, rennet-induced gelation, including rennet coagulation time, gel strength, and yield stress, was measured in skim milk samples collected from 379 animals of the Swedish Red breed using low-amplitude oscillation measurements. All animals had genotypes for almost 621,000 segregating single nucleotide polymorphisms (SNP), identified using the Bovine HD SNPChip (Illumina Inc., San Diego, CA). The genome was scanned for associations, haplotypes based on SNP sets comprising highly associated SNP were inferred, and the effects of the 2 most common haplotypes within each region were analyzed using mixed models. Even though the number of animals was relatively small, a total of 21 regions were identified, with 4 regions showing association with more than one trait. A major quantitative trait locus for all traits was identified around the casein cluster explaining between 9.3 to 15.2% of the phenotypic variation of the different traits. In addition, 3 other possible candidate genes were identified; that is, UDP-N-acetyl-α-d-galactosamine:polypeptide N-acetylgalactosaminyl-transferase 1 (GALNT1), playing a role in O-glycosylation of κ-casein, and 2 cathepsins, CTSZ and CTSC, possibly involved in proteolysis of milk proteins. We have shown that other genes than the casein genes themselves may be involved in the regulation of gelation traits. However, additional analysis is needed to confirm these results. To our knowledge, this is the first study identifying quantitative trait loci affecting rennet-induced gelation of skim milk through a high-density genome-wide association study.
Assuntos
Bovinos/genética , Quimosina , Géis/química , Leite/química , Reologia , Animais , Cruzamento , Caseínas/genética , Queijo , Fenômenos Químicos , Mapeamento Cromossômico/veterinária , Feminino , Genoma , Estudo de Associação Genômica Ampla , Genótipo , Haplótipos/genética , Lactoglobulinas/genética , Leite/metabolismo , Proteínas do Leite/metabolismo , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Locos de Características Quantitativas/genética , Reprodutibilidade dos Testes , ViscosidadeRESUMO
Milk coagulation is an important processing trait, being the basis for production of both cheese and fermented products. There is interest in including technological properties of these products in the breeding goal for dairy cattle. The aim of the present study was therefore to estimate genetic parameters for milk coagulation properties, including both rennet- and acid-induced coagulation, in Swedish Red dairy cattle using genomic relationships. Morning milk samples and blood samples were collected from 395 Swedish Red cows that were selected to be as genetically unrelated as possible. Using a rheometer, milk samples were analyzed for rennet- and acid-induced coagulation properties, including gel strength (G'), coagulation time, and yield stress (YS). In addition to the technological traits, milk composition was analyzed. A binary trait was created to reflect that milk samples that had not coagulated 40min after rennet addition were considered noncoagulating milk. The cows were genotyped by using the Illumina BovineHD BeadChip (Illumina Inc., San Diego, CA). Almost 600,000 markers remained after quality control and were used to construct a matrix of genomic relationships among the cows. Multivariate models including fixed effects of herd, lactation stage, and parity were fitted using the ASReml software to obtain estimates of heritabilities and genetic and phenotypic correlations. Heritability estimates (h(2)) for G' and YS in rennet and acid gels were found to be high (h(2)=0.38-0.62) and the genetic correlations between rennet-induced and acid-induced coagulation properties were weak but favorable, with the exception of YSrennet with G'acid and YSacid, both of which were strong. The high heritability (h(2)=0.45) for milk coagulating ability expressed as a binary trait suggests that noncoagulation could be eliminated through breeding. Additionally, the results indicated that the current breeding objective could increase the frequency of noncoagulating milk and lead to deterioration of acid-induced coagulation through unfavorable genetic associations with protein content (0.38) and milk yield (-0.61 to -0.71), respectively. The outcome of this study suggests that by including more detailed compositional traits genetically associated with milk coagulation or by including milk coagulation properties directly within the breeding goal, it appears possible to breed cows that produce milk better suited for production of cheese and fermented products.
Assuntos
Bovinos/genética , Quimosina/genética , Leite/química , Animais , Cruzamento , Caseínas/genética , Bovinos/fisiologia , Quimosina/metabolismo , Feminino , Genótipo , Lactação/genética , Leite/metabolismo , Paridade , Fenótipo , Gravidez , SuéciaRESUMO
In selecting cows for higher milk yields and milk quality, it is important to understand how these traits are affected by the bovine genome. The major milk proteins exhibit genetic polymorphism and these genetic variants can serve as markers for milk composition, milk production traits, and technological properties of milk. The aim of this study was to investigate the relationships between casein (CN) genetic variants and detailed protein composition in Swedish and Danish dairy milk. Milk and DNA samples were collected from approximately 400 individual cows each of 3 Scandinavian dairy breeds: Swedish Red (SR), Danish Holstein (DH), and Danish Jersey (DJ). The protein profile with relative concentrations of α-lactalbumin, ß-lactoglobulin, and α(S1)-, α(S2)-, κ-, and ß-CN was determined for each milk sample using capillary zone electrophoresis. The genetic variants of the α(S1)- (CSN1S1), ß- (CSN2), and κ-CN (CSN3) genes for each cow were determined using TaqMan SNP genotyping assays (Applied Biosystems, Foster City, CA). Univariate statistical models were used to evaluate the effects of composite genetic variants, α(S1)-ß-κ-CN, on the protein profile. The 3 studied Scandinavian breeds differed from each other regarding CN genotypes, with DH and SR having similar genotype frequencies, whereas the genotype frequencies in DJ differed from the other 2 breeds. The similarities in genotype frequencies of SR and DH and differences compared with DJ were also seen in milk production traits, gross milk composition, and protein profile. Frequencies of the most common composite α(S1)-ß-κ-CN genotype BB/A(2)A(2)/AA were 30% in DH and 15% in SR, and cows that had this genotype gave milk with lower relative concentrations of κ- and ß-CN and higher relative concentrations of αS-CN, than the majority of the other composite genotypes in SR and DH. The effect of composite genotypes on relative concentrations of the milk proteins was not as pronounced in DJ. The present work suggests that a higher frequency of BB/A(1)A(2)/AB, together with a decrease in BB/A(2)A(2)/AA, could have positive effects on DH and SR milk regarding, for example, the processing of cheese.
Assuntos
Caseínas/genética , Bovinos/genética , Proteínas do Leite/genética , Leite/química , Polimorfismo Genético , Animais , Caseínas/metabolismo , Bovinos/metabolismo , Dinamarca , Feminino , Genótipo , Lactalbumina/genética , Lactoglobulinas/genética , Lactoglobulinas/metabolismo , Proteínas do Leite/análise , Proteínas do Leite/metabolismo , Especificidade da Espécie , SuéciaRESUMO
Treatment of Caco-2 cells with the peptide lactoferricin(4-14), results in reduction of the growth rate by prolongation of the S phase of the cell cycle. Lactoferricin(1-25) is formed in the gut by cleavage from lactoferrin and the bioactive amino acids are found within lactoferricin(4-14). Our hypothesis is that the reduction of the rate of S phase progression may result in increased DNA repair. To test this hypothesis, Caco-2 cells were subjected to UV light that caused DNA lesions and then the cells were grown in the absence or presence of 2.0 µM lactoferricin(4-14). Evaluation of DNA strand breaks using the comet assay showed that lactoferricin(4-14) treatment indeed resulted in a reduction of comets showing damaged DNA. In the search for a mechanism, we have investigated the levels of several proteins involved in cell cycle regulation, DNA replication, and apoptosis using Western blot. Lactoferricin(4-14) treatment resulted in an increased expression of flap endonuclease-1 pointing to increased DNA synthesis activity. Lactoferricin(4-14) treatment decreased the expression of the proapoptotic protein B-cell lymphoma 2-associated X protein (or Bax), indicating decreased cell death. As we have found previously, lactoferricin(4-14) treatment reduced the expression of cyclin E involved in the G(1)/S transition. Immunofluorescence microscopy showed that a lower γ-H2AX expression in lactoferricin(4-14)-treated cells, pointing to more efficient DNA repair. Thus, altogether our data show that lactoferricin(4-14) treatment has beneficial effects.
Assuntos
Antineoplásicos/farmacologia , Células CACO-2/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Lactoferrina/farmacologia , Fragmentos de Peptídeos/farmacologia , Western Blotting , Linhagem Celular Tumoral , Ensaio Cometa/métodos , Reparo do DNA/efeitos dos fármacos , Humanos , Fase S/efeitos dos fármacos , Raios Ultravioleta/efeitos adversosRESUMO
Cheese production has increased worldwide during the last decade and is expected to increase within the coming decade as well. Despite this, the relations between cow genetics and cheese characteristics are not fully known. The aim of this study was to determine if polymorphisms in the leptin (LEP), leptin receptor (LEPR), and acyl-coenzyme A:diacylglycerol acyltransferase 1 (DGAT1) genes as well as genetic variants of ß-casein (ß-CN), κ-CN, and ß-lactoglobulin (ß-LG) affect technological properties important for cheese production and, hence, could act as genetic makers for cheese quality. Individual milk samples from the Swedish Red and the Swedish Holstein breeds were analyzed for sizes of CN micelles and fat globules as well as rennet-induced gel strength, gelation time, and yield stress. Model cheeses were produced to study yield, hardness, and pH of the cheeses. The A1457G, A252T, A59V, and C963T single nucleotide polymorphisms (SNP) were analyzed on the LEP gene, the T945M SNP on the LEPR gene, and the Nt984+8(A-G) SNP on the DGAT1 gene. In addition, genetic variants of ß-CN, κ-CN, and ß-LG were determined. The results indicate that technological properties were influenced by the LEPR(T945M) polymorphism, which had an association with gel strength, yield stress, and cheese hardness (T > C). However, also LEP(A252T) was shown to affect gel strength (T > A), whereas the LEP(A59V) had an effect on fat globule size (T > C). For the milk protein genes, favorable effects were found for the A and B variants of ß-LG and κ-CN, respectively, on gel strength, gelation time, and yield stress. In addition, the B variant of κ-CN was shown to be associated with smaller CN micelles than the A variant. Thus, the results demonstrate potential genetic markers for cheese characteristics. However, milk composition traits also affected the obtained results, thus making it necessary to thoroughly assess the different aspects regarding the influence of gene effects on cheese characteristics before directly selecting for certain alleles or genetic variants to improve the processing and quality of cheese.
Assuntos
Queijo/análise , Diacilglicerol O-Aciltransferase/genética , Leptina/genética , Proteínas do Leite/genética , Polimorfismo Genético , Receptores para Leptina/genética , Animais , Caseínas/genética , Marcadores Genéticos , Lactoglobulinas/genéticaRESUMO
Coagulation properties of milk are altered by elevated somatic cell count (SCC), partly due to increased proteolytic and lipolytic activity in the milk and, thereby, degradation of protein and fat during storage. Milk is commonly stored on the farm at cooling conditions for up to 2 d before transport to the dairy for processing. This study evaluated the effects of storage on milk with altered composition due to high SCC and the effects of exclusion of milk from individual udder quarters with high SCC on milk composition, proteolysis, and coagulation properties. Udder-quarter milk and cow-composite milk samples from 13 cows having at least 1 quarter with SCC above 100,000 cells/mL were collected on 1 occasion. In addition, commingled milk from only healthy quarters (<100,000 cells/mL) of each cow was collected, representing a cow sample where milk with elevated SCC was excluded. The milk samples were analyzed for total protein content; protein content in the whey fraction; casein, fat, and lactose contents; SCC; proteolysis; curd yield; coagulation time; and total bacterial count, on the day of sampling and after 2 and 5 d of storage at +4°C. In addition to SCC, duration of storage and total bacterial count had an effect on milk quality. The content of total protein, fat and protein contents in the whey fraction, and curd yield were found to have different storage characteristics depending on the level of SCC at udder-quarter level. The exclusion of milk from udder quarters with elevated SCC decreased the content of total protein and protein content in the whey fraction and increased the content of lactose at cow level. However, the effect of separating milk at udder-quarter level needs to be further studied at bulk tank level to evaluate the effect on overall total milk quality.
Assuntos
Indústria de Laticínios , Manipulação de Alimentos , Leite/química , Leite/citologia , Animais , Gorduras/análise , Lactose/análise , Leite/microbiologia , Proteínas do Leite/análise , Proteólise , Fatores de TempoRESUMO
The main objective of this study was to investigate whether the α-lactalbumin (α-LA) content of bulk milk is related with some known inflammatory markers and milk quality traits. An additional objective was to study whether combining α-LA, haptoglobin (Hp), and serum amyloid A (SAA) in an acute phase index (API) could be useful as an alternative marker for bulk milk quality. For the dairy industry, it is of great importance to receive high quality bulk milk for manufacture of liquid milk and dairy products. The somatic cell count (SCC) is currently used as an indirect marker for bulk milk quality, but because it is somewhat insensitive and unspecific, interest exists in alternative markers. Bulk milk samples were analyzed for α-LA, SCC, polymorphonuclear leukocyte count, Hp, SAA, fat, lactose, total protein and casein contents, casein number, protein composition, proteolysis, and coagulating properties. No significant differences were found in SCC, polymorphonuclear leukocyte count, Hp, or SAA between milk samples containing low, medium, or high concentrations of α-LA. Differences between α-LA groups were, however, found in some milk quality traits because high α-LA concentration was related to low concentrations of α(S1)-, α(S2)-, and ß-caseins and high concentrations of lactose and ß-lactoglobulin. A high API was related to low lactose content and casein number. Samples with high SCC contained less casein and had a lower casein number than milk with a low SCC, and proteolysis was significantly higher in high SCC milk than in low SCC milk. Neither α-LA nor API proved to be a better marker than SCC for the quality traits investigated, and α-LA was not considered to be a useful inflammatory marker in bulk milk.
Assuntos
Lactalbumina/análise , Mastite Bovina/diagnóstico , Leite/química , Animais , Biomarcadores/análise , Bovinos , Contagem de Células , Feminino , Haptoglobinas/análise , Leite/normas , Controle de Qualidade , Proteína Amiloide A Sérica/análiseRESUMO
Bovine milk is associated with improved health and reduced risk of several diseases, among them cancer. Milk is a complex mixture of known and unknown components. The components and the mechanisms that contribute to the cancer-preventive effects are largely unknown. We set out to find new peptides in milk and identified ubiquitin (Ub) using matrix-assisted laser desorption ionization-time of flight mass spectrometry and Western blot. Using quantitative Western blot, we estimated the Ub concentration to be about 0.003 micromol/L in milk. We then decided to investigate the effect of treating human colon cancer CaCo-2 cells with Ub, using higher concentrations than in milk. CaCo-2 cells treated with 0.02 to 2.0 micromol/L Ub showed significantly decreased proliferation compared with untreated control cells. A higher growth inhibitory effect than in CaCo-2 cells was found in the neuroblastoma cell line SH-SY5Y treated with 0.02 to 0.2 micromol/L Ub. A bromodeoxyuridine DNA flow cytometric method was used to study cell cycle kinetics in Ub-treated CaCo-2 cells. The data point toward a prolongation of the G(1) phase. The levels of several cell cycle regulatory proteins were affected. Our data point to Ub possibly being one of the components in milk reducing the risk of cancer.
Assuntos
Inibidores do Crescimento/farmacologia , Leite/química , Ubiquitina/farmacologia , Animais , Células CACO-2 , Bovinos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Inibidores do Crescimento/análise , Humanos , Ubiquitina/análiseRESUMO
Automatic in-line measurement of milk composition and milk yield could be a useful tool in management of the dairy herd. Data on milk components and milk yield provide information on milk quality alterations and cow health status but are also useful in planning feeding and breeding. In automatic milking systems, udder quarters are milked individually, enabling analysis and recording at the udder-quarter level. Frequent records of components require knowledge about day-to-day variations. A component with greater day-to-day variation needs more frequent sampling when used as a diagnostic tool and for management decisions. Earlier studies have described the day-to-day variations in milk components for cow composite milk, but with the quarter milking technique and the possible sampling at the udder-quarter level, knowledge about day-to-day variations at the udder-quarter level is needed. In this study, udder-quarter and cow composite milk samples were collected from 42 consecutive milkings of 10 cows during 21 d. Milk yield was recorded and the milk was analyzed for total protein, whey protein, casein, fat, lactose, and somatic cell count. The results showed that the day-to-day variations and mean values for 4 healthy udder quarters within a cow were similar. In addition, different milk components had different levels of day-to-day variation, the least variation being found in lactose (0.9%) and the greatest in fat (7.7%). This suggests that repeated milk sampling and analysis at the udder-quarter level can be used to detect alterations in composition and cow health and would, thus, be helpful in the management of the dairy herd.
Assuntos
Bovinos/fisiologia , Indústria de Laticínios/métodos , Leite/química , Leite/metabolismo , Animais , Caseínas/análise , Contagem de Células/veterinária , Feminino , Lactação , Lactose/análise , Lipídeos/análise , Proteínas do Leite/análise , Fatores de Tempo , Proteínas do Soro do LeiteRESUMO
The economic output of the dairy industry is to a great extent dependent on the processing of milk into other milk-based products such as cheese. The yield and quality of cheese are dependent on both the composition and technological properties of milk. The objective of this study was to evaluate the importance and effects of casein (CN) micelle size and milk composition on milk gelation characteristics in order to evaluate the possibilities for enhancing gelation properties through breeding. Milk was collected on 4 sampling occasions at the farm level in winter and summer from dairy cows with high genetic merit, classified as elite dairy cows, of the Swedish Red and Swedish Holstein breeds. Comparisons were made with milk from a Swedish Red herd, a Swedish Holstein herd, and a Swedish dairy processor. Properties of CN micelles, such as their native and rennet-induced CN micelle size and their zeta-potential, were analyzed by photon correlation spectroscopy, and rennet-induced gelation characteristics, including gel strength, gelation time, and frequency sweeps, were determined. Milk parameters of the protein, lipid, and carbohydrate profiles as well as minerals were used to obtain correlations with native CN micelle size and gelation characteristics. Milk pH and protein, CN, and lactose contents were found to affect milk gelation. Smaller native CN micelles were shown to form stronger gels when poorly coagulating milk was excluded from the correlation analysis. In addition, milk pH correlated positively, whereas Mg and K correlated negatively with native CN micellar size. The milk from the elite dairy cows was shown to have good gelation characteristics. Furthermore, genetic progress in relation to CN micelle size was found for these cows as a correlated response to selection for the Swedish breeding objective if optimizing for milk gelation characteristics. The results indicate that selection for smaller native CN micelles and lower milk pH through breeding would enhance gelation properties and may thus improve the initial step in the processing of cheese.
Assuntos
Cruzamento , Caseínas/análise , Bovinos/fisiologia , Indústria de Laticínios/métodos , Leite/química , Animais , Bovinos/genética , Indústria de Laticínios/economia , Feminino , Géis , Lactação/genética , Lactação/metabolismo , Micelas , Estações do Ano , Seleção GenéticaRESUMO
Food components modify the risk of cancer at a large number of sites but the mechanism of action is unknown. In the present investigation, we studied the effect of the peptide lactoferricin derived from bovine milk lactoferrin on human colon cancer CaCo-2 cells. The cells were either untreated or treated with 2.0, 0.2, or 0.02 microM lactoferricin. Cell cycle kinetics were investigated with a bromodeoxyuridine DNA flow cytometric method. The results show that lactoferricin treatment slightly but significantly prolonged the S phase of the cell cycle. Lactoferricin treatment lowered the level of cyclin E1, a protein involved in the regulation of genes required for G(1)/S transition and consequently for efficient S phase progression. The slight prolongation of the S phase resulted in a reduction of cell proliferation, which became more apparent after a long treatment time.
Assuntos
Antibacterianos/farmacologia , Lactoferrina/farmacologia , Antibacterianos/toxicidade , Células CACO-2 , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Lactoferrina/toxicidade , Fase S/efeitos dos fármacos , Fatores de TempoRESUMO
The objective of this work was to find regressions between minor milk proteins or protein fragments in the casein or sweet whey fraction and cheese yield because the effect of major milk proteins was evaluated in a previous study. Proteomic methods involving 2-dimensional gel electrophoresis and mass spectrometry in combination with multivariate data analysis were used to study the effect of variations in milk protein composition in chymosin separated whey and casein fractions on cheese yield. By mass spectrometry, a range of proteins significant for the cheese yield was identified. Among others, a C-terminal fragment of beta-casein had a positive effect on the cheese yield expressed as grams of cheese per 100 g of milk, whereas several other minor fragments of beta-, alpha(s1)-, and alpha(s2)-casein had positive effects on the transfer of protein from milk to cheese. However, the individual effect of each identified protein was relatively low. Therefore, further studies of the relations between different proteins/peptides in the rennet casein or sweet whey fractions and cheese yield are needed for advanced understanding and prediction of cheese yield.
Assuntos
Caseínas/análise , Queijo/análise , Quimosina/metabolismo , Proteínas do Leite/análise , Animais , Eletroforese em Gel Bidimensional , Análise dos Mínimos Quadrados , Espectrometria de Massas , Leite/química , Análise Multivariada , Proteômica , Proteínas do Soro do LeiteRESUMO
The objective of this study was to evaluate the effect of variations in milk protein composition on milk clotting properties and cheese yield. Milk was collected from 134 dairy cows of Swedish Red and White, Swedish Holstein, and Danish Holstein-Friesian breed at 3 sampling occasions. Concentrations of alphaS1-, beta-, and kappa-casein (CN), alpha-lactalbumin, and beta-lactoglobulin (LG) A and B were determined by reversed phase liquid chromatography. Cows of Swedish breeds were genotyped for genetic variants of beta- and kappa-CN. Model cheeses were produced from individual skimmed milk samples and the milk clotting properties were evaluated. More than 30% of the samples were poorly coagulating or noncoagulating, resulting in weak or no coagulum, respectively. Poorly and noncoagulating samples were associated with a low concentration of kappa-CN and a low proportion of kappa-CN in relation to total CN analyzed. Furthermore, the kappa-CN concentration was higher in milk from cows with the AB genotype than the AA genotype of kappa-CN. The concentrations of alphaS1-, beta-, and kappa-CN and of beta-LG B were found to be significant for the cheese yield, expressed as grams of cheese per one hundred grams of milk. The ratio of CN to total protein analyzed and the beta-LG B concentration positively affected cheese yield, expressed as grams of dry cheese solids per one hundred grams of milk protein, whereas beta-LG A had a negative effect. Cheese-making properties could be improved by selecting milk with high concentrations of alphaS1-, beta-, and kappa-CN, with high kappa-CN in relation to total CN and milk that contains beta-LG B.
Assuntos
Caseínas/genética , Queijo , Proteínas do Leite/química , Animais , Caseínas/análise , Bovinos , Queijo/normas , Indústria de Laticínios , Feminino , Genótipo , Lactalbumina/análise , Lactoglobulinas/análise , Análise dos Mínimos Quadrados , Leite/química , Proteínas do Leite/genética , Fatores de TempoRESUMO
OBJECTIVE: To assess the absorption of dietary selenium in humans, especially of milk selenium. DESIGN: : 1-day meal studies in subjects with ileostomy. SETTING: Hospital outpatient clinics. SUBJECTS: Three subjects in the pilot study and nine subjects in the main study (eight men/ four women). INTERVENTION: Different beverages, 1 l/day, were given in addition to basal diets (soft drink, 1 week; low-fat milk, 3 weeks; fermented low-fat milk, 3 weeks and soft drink, 1 week). Ileostomy effluents were collected during the last 2 days in each of the four periods. RESULTS: On days when the subjects were given 1 l of low-fat milk, the estimated fractional absorption of total dietary selenium was 65.5 (2.3)% (mean (s.d.), n=18), which was similar to the value when fermented low-fat milk was given (64.1 (3.2)%). However, both the calculated amount of milk selenium absorbed (10.9 (2.4) vs 9.4 (1.7) microg selenium) and its fractional absorption (73.3 (16.1) vs 64.1 (11.2)%, n=18) were significantly higher for milk than for fermented milk. CONCLUSIONS: Selenium from milk and other sources is well absorbed in subjects with ileostomy. The real absorption may be even higher than the values shown.
Assuntos
Gorduras na Dieta , Ileostomia , Leite/metabolismo , Selênio/farmacocinética , Adulto , Animais , Disponibilidade Biológica , Produtos Fermentados do Leite/metabolismo , Dieta , Feminino , Humanos , Absorção Intestinal , Masculino , Pessoa de Meia-Idade , Leite/química , Projetos Piloto , Selênio/administração & dosagem , SuéciaRESUMO
Much emphasis has been put on evaluating alterations in milk composition caused by clinical and subclinical mastitis. However, little is known about changes in milk composition during subclinical mastitis in individual udder quarters with a low-to-moderate increase in milk somatic cell count (SCC). This information is needed to decide whether milk from individual udder quarters with a moderate-to-high increase in milk SCC should be separated or not. The aim of this study was to determine how milk composition in separate udder quarters is affected when cow composite milk has low or moderately increased SCC levels. Udder quarter and cow composite milk samples were collected from 17 cows on one occasion. Milk yield was registered and samples were analyzed for SCC, fat, total protein, whey proteins, lactose, citric acid, non-protein nitrogen (NPN), lactoferrin, protein profile, free fatty acids (FFAs), lactate dehydrogenase (LDH), proteolysis, sodium and potassium. Bacteriological samples were collected twice from all four quarters of all cows. The cows were divided into three groups depending on their SCC at udder quarter level. The first group comprised healthy cows with four udder quarters with low SCC, <50 000 cells/ml; composition was equal when opposite rear and front quarters were compared. In the second and the third groups, cows had one udder quarter with 101 000 cells/ml < SCC < 600 000 cells/ml and SCC > 700 000 cells/ml, respectively. The remaining udder quarters of these cows had low SCC (<100 000 cells/ml). Despite the relatively low average cow composite SCC = 100 000 cells/ml of Group 2, milk from affected udder quarters exhibited lower casein number, content of lactose and ß-casein (ß-CN), while the content of whey protein, sodium, LDH and α-lactoalbumin (α-la) were higher compared to healthy opposite quarters. In addition to these changes, milk from affected udder quarters in Group 3 also exhibited lower values of potassium and αs1-casein (αs1-CN) and higher values of lactoferrin when compared to milk from opposite healthy quarters. This indicates that even when the SCC in cow composite milk is low, there might exist individual quarters for which milk composition is changed and milk quality impaired.
RESUMO
Automatic milking systems have made possible the separation of high- and low-quality milk at the udder quarter level during the milking process. The aim of this study was to investigate the composition and yield of milk from individual udder quarters to determine whether deteriorated milk composition occurs in udders that are assumed to be healthy and whether quarters with high-quality milk are found in udders with high milk somatic cell count (SCC). Milk samples were collected on one occasion from 90 cows at udder quarter level and cow composite level. The milk was analyzed for content of total protein, whey protein, casein, fat, lactose, citric acid and SCC; milk yield was registered. The cows were divided into three groups depending on the SCC of their composite milk. Cows in group 1, cow composite SCC < 100 000 cells/ml, were assumed to have healthy udders. However, instances of increased SCC and decreased milk quality were discovered in one or more udder quarters of approximately 30% of the group. Cows in group 2, cow composite SCC of 100 000 to 300 000 cells/ml, and group 3, cow composite SCC > 300 000 cells/ml, were assumed to have affected udders. However, the majority of these cows had one or more udder quarters in which increased SCC and deteriorated milk quality were not detected. Calculations of bulk-tank milk values, when separation of milk from affected udder quarters was performed, indicate that SCC changes to a much greater degree compared to the other milk components. These results show that milk from affected udder quarters suffers compositional changes, but calculations of simulated separation indicate that the compositional changes in bulk-tank milk are small. The effect of separation of milk from individual udder quarters on bulk-tank milk needs to be further studied.
RESUMO
Lipid auto-oxidation in milk is affected by a complex interplay of pro- and antioxidants. Several of these compounds are also important nutrients in the human diet and may have other physiological effects in the gastrointestinal tract and other tissues. Among antioxidative enzymes superoxide dismutase catalyses the dismutation of superoxide anion to hydrogen peroxide. The degradation of hydrogen peroxide can be catalysed by catalase and the selenoprotein glutathione peroxidase. The latter enzyme can also degrade lipid peroxides. Lactoferrin may have an important role by binding pro-oxidative iron ions. The occurrence of different forms of these antioxidative proteins in milk and available data on their functional role are reviewed. More remains to be learnt of individual compounds and as an example the potential role of seleno compounds in milk is virtually unknown. Antioxidative vitamins in milk can provide an important contribution to the daily dietary intake. Moreover vitamin E and carotenoids act as fat-soluble antioxidants, e.g. in the milk fat globule membrane, which is regarded as a major site of auto-oxidation. Vitamin C is an important water-soluble antioxidant and interacts in a complex manner with iron and fat-soluble antioxidants. The concentrations of these compounds in milk are affected by cow feeding rations and milk storage conditions. Since milk contains a number of antioxidants many reactions are possible and the specific function of each antioxidant cannot easily be defined. There are indications that other compounds may have antioxidative function and measurement of total antioxidative capacity should be a useful tool in evaluating their relative roles.