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Sample records for bovine milk xanthine

  1. Subunit structure of bovine milk xanthine oxidase. Effect of limited cleavage by proteolytic enzymes on activity and structure.

    Science.gov (United States)

    Nagler, L G; Vartanyan, L S

    1976-03-18

    Bovine milk xanthine oxidase (xanthine:oxygen oxidoreductase, EC 1.2.3.2) has been purified by a modified method without the use of proteases, and its structure has been analyzed by polyacrylamide gel electrophoresis. Native xanthine oxidase is found to consist of only two polypeptide chains A with molecular weights of 150 000 each. These chains have NH2-terminal methionine. Limited proteolysis with trypsin, chymotrypsin, or subtilisin at pH 8 did not affect molecular weight and activities of the enzyme while each of the A chains was cleaved under these conditions to three fragments C, E, and F with molecular weights of 92 00, 42 000 and 20 000, respectively. These fragments remained bound to each other and were relatively resistant to subsequent proteolysis. The isolation of xanthine oxidase in the presence of pancreatin as described by Hart et al. (1970, Biochem. J. 116, 851) gives partially digested enzyme composed mainly of chains C, E (Mr 35 000) and a small component (Mr approx. 15 0-0). The action of subtilisin on xanthine oxidase at pH 11 resulted in complete digestion of E chains, FAD separation, and total loss of xanthine:oxygen oxidoreductase activity while xanthine:indophenol oxidoreductase activity was relatively little affected. The residual enzyme has a molecular weight of about 200 000, is composed mainly of two C chains (and may probably contain F and/or proteolytic fragments of low molecular weight), contains molybdenum, and does not contain FAD.

  2. Thermal properties of milk fat, xanthine oxidase, caseins and whey proteins in pulsed electric field-treated bovine whole milk.

    Science.gov (United States)

    Sharma, Pankaj; Oey, Indrawati; Everett, David W

    2016-09-15

    Thermodynamics of milk components (milk fat, xanthine oxidase, caseins and whey proteins) in pulsed electric field (PEF)-treated milk were compared with thermally treated milk (63 °C for 30 min and 73 °C for 15s). PEF treatments were applied at 20 or 26 kV cm(-1) for 34 μs with or without pre-heating of milk (55 °C for 24s), using bipolar square wave pulses in a continuous mode of operation. PEF treatments did not affect the final temperatures of fat melting (Tmelting) or xanthine oxidase denaturation (Tdenaturation), whereas thermal treatments increased both the Tmelting of milk fat and the Tdenaturation for xanthine oxidase by 2-3 °C. Xanthine oxidase denaturation was ∼13% less after PEF treatments compared with the thermal treatments. The enthalpy change (ΔH of denaturation) of whey proteins decreased in the treated-milk, and denaturation increased with the treatment intensity. New endothermic peaks in the calorimetric thermograms of treated milk revealed the formation of complexes due to interactions between MFGM (milk fat globule membrane) proteins and skim milk proteins. Evidence for the adsorption of complexes onto the MFGM surface was obtained from the increase in surface hydrophobicity of proteins, revealing the presence of unfolded hydrophobic regions.

  3. Aldehyde-induced xanthine oxidase activity in raw milk.

    Science.gov (United States)

    Steffensen, Charlotte L; Andersen, Henrik J; Nielsen, Jacob H

    2002-12-04

    In the present study, the aldehyde-induced pro-oxidative activity of xanthine oxidase was followed in an accelerated raw milk system using spin-trap electron spin resonance (ESR) spectroscopy. The aldehydes acetaldehyde, propanal, hexanal, trans-2-hexenal, trans-2-heptenal, trans-2-nonenal, and 3-methyl-2-butenal were all found to initiate radical reactions when added to milk. Formation of superoxide through aldehyde-induced xanthine oxidase activity is suggested as the initial reaction, as all tested aldehydes were shown to trigger superoxide formation in an ultrahigh temperature (UHT) milk model system with added xanthine oxidase. It was found that addition of aldehydes to milk initially increased the ascorbyl radical concentration with a subsequent decay due to ascorbate depletion, which renders the formation of superoxide in milk with added aldehyde. The present study shows for the first time potential acceleration of oxidative events in milk through aldehyde-induced xanthine oxidase activity.

  4. Bovine milk glycome.

    Science.gov (United States)

    Tao, N; DePeters, E J; Freeman, S; German, J B; Grimm, R; Lebrilla, C B

    2008-10-01

    Bovine milk oligosaccharides have several potentially important biological activities including the prevention of pathogen binding to the intestinal epithelial and as nutrients for beneficial bacteria. It has been suggested that milk oligosaccharides are an important source of complex carbohydrates as supplements for the food and the pharmaceutical industries. However, only a small number of structures of bovine milk oligosaccharides (bMO) are known. There have been no systematic studies on bMO. High-performance mass spectrometry and separation methods are used to evaluate bMO, and nearly 40 oligosaccharides are present in bovine milk. Bovine milk oligosaccharides are composed of shorter oligomeric chains than are those in human milk. They are significantly more anionic with nearly 70%, measured abundances, being sialylated. Additionally, bMO are built not only on the lactose core (as are nearly all human milk oligosaccharides), but also on lactose amines. Sialic acid residues include both N-acetyl and N-glycolylneuraminic acid, although the former is significantly more abundant.

  5. Bovine milk exosome proteome

    Science.gov (United States)

    Exosomes are 40-100 nm membrane vesicles of endocytic origin and are found in blood, urine, amniotic fluid, bronchoalveolar lavage (BAL) fluid, as well as human and bovine milk. Exosomes are extracellular organelles important in intracellular communication/signaling, immune function, and biomarkers ...

  6. Proteome profile and biological activity of caprine, bovine and human milk fat globules.

    Science.gov (United States)

    Spertino, Stefano; Cipriani, Valentina; De Angelis, Chiara; Giuffrida, Maria Gabriella; Marsano, Francesco; Cavaletto, Maria

    2012-04-01

    Upon combining bidimensional electrophoresis with monodimensional separation, a more comprehensive analysis of the milk fat globule membrane has been obtained. The proteomic profile of caprine milk fat globules revealed the presence of butyrophilin, lactadherin and perilipin as the major proteins, they were also associated to bovine and human milk fat globule membranes. Xanthine dehydrogenase/oxidase has been detected only in monodimensional gels. Biological activity of milk fat globules has been evaluated in Caco2-cells, as a representative model of the intestinal barrier. The increase of cell viability was indicative of a potential nutraceutical role for the whole milk fat globule, suggesting a possible employment in milk formula preparation.

  7. Bovine milk antibodies for health.

    Science.gov (United States)

    Korhonen, H; Marnila, P; Gill, H S

    2000-11-01

    The immunoglobulins of bovine colostrum provide the major antimicrobial protection against microbial infections and confer a passive immunity to the newborn calf until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunising cows with these pathogens or their antigens. Immune milk products are preparations made of such hyperimmune colostrum or antibodies enriched from it. These preparations can be used to give effective specific protection against different enteric diseases in calves and suckling pigs. Colostral immunoglobulin supplements designed for farm animals are commercially available in many countries. Also, some immune milk products containing specific antibodies against certain pathogens have been launched on the market. A number of clinical studies are currently in progress to evaluate the efficacy of immune milks in the prevention and treatment of various human infections, including those caused by antibiotic resistant bacteria. Bovine colostrum-based immune milk products have proven effective in prophylaxis against various infectious diseases in humans. Good results have been obtained with products targeted against rotavirus, Shigella flexneri, Escherichia coli, Clostridium difficile, Streptococcus mutans, Cryptosporidium parvum and Helicobacter pylori. Some successful attempts have been made to use immune milk in balancing gastrointestinal microbial flora. Immune milk products are promising examples of health-promoting functional foods, or nutraceuticals. This review summarises the recent progress in the development of these products and evaluates their potential as dietary supplements and in clinical nutrition.

  8. Studies on the quantitative structure-activity relationship of the inhibition of xanthine oxidase by azaheterocyclic compounds.

    NARCIS (Netherlands)

    Naeff, H.S.D.

    1990-01-01

    This thesis contains the results of a QSAR analysis of the interaction of bovine milk xanthine oxidase with two azaheterocyclic compounds, namely the 6-arylpteridin- 4-ones and the 8-arylhypoxanthines. Xanthine oxidase has active sites for various substrates. The studies done for this thesis were of

  9. Potential Anticarcinogenic Peptides from Bovine Milk

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    Giacomo Pepe

    2013-01-01

    Full Text Available Bovine milk possesses a protein system constituted by two major families of proteins: caseins (insoluble and whey proteins (soluble. Caseins (αS1, αS2, β, and κ are the predominant phosphoproteins in the milk of ruminants, accounting for about 80% of total protein, while the whey proteins, representing approximately 20% of milk protein fraction, include β-lactoglobulin, α-lactalbumin, immunoglobulins, bovine serum albumin, bovine lactoferrin, and lactoperoxidase, together with other minor components. Different bioactivities have been associated with these proteins. In many cases, caseins and whey proteins act as precursors of bioactive peptides that are released, in the body, by enzymatic proteolysis during gastrointestinal digestion or during food processing. The biologically active peptides are of particular interest in food science and nutrition because they have been shown to play physiological roles, including opioid-like features, as well as immunomodulant, antihypertensive, antimicrobial, antiviral, and antioxidant activities. In recent years, research has focused its attention on the ability of these molecules to provide a prevention against the development of cancer. This paper presents an overview of antitumor activity of caseins and whey proteins and derived peptides.

  10. Fluorometric determination of uric acid in bovine milk

    DEFF Research Database (Denmark)

    Larsen, Torben; Moyes, Kasey

    2010-01-01

    The primary objective of this study is to validate a new fast method for determination of uric acid in milk. The method is based on an enzymatic-fluorometric technique that requires minimal pre-treatment of milk samples. The present determination of uric acid is based on the enzymatic oxidation...... at -18°C for 24 h. Addition of either allopurinol (a xanthine oxidase inhibitor) or dimethylsulfoxide (a solvent for allopurinol) did not affect milk uric acid concentrations (P=0·96) and may indicate that heat treatment before storage and analysis was sufficient to degrade xanthine oxidase activity...... of uric acid to 5-hydroxyisourate via uricase where the liberated hydrogen peroxide reacts with 10-acetyl-3,7-dihydroxyphenoxazine via peroxidase and the fluorescent product, resorufin, is measured fluorometrically. Fresh composite milk samples (n=1,072) were collected from both Jersey (n=38) and Danish...

  11. Genetic analysis of protein composition of bovine milk

    NARCIS (Netherlands)

    Schopen, G.C.B.

    2010-01-01

    This thesis is part of the Dutch Milk Genomics Initiative, and the general aim was to obtain more insight into the genetic background of bovine milk protein composition. Morning milk samples from roughly 2000 cows were analyzed for the six major milk proteins (αS1-casein, αS2-casein, β-casein, κ-cas

  12. The host defense proteome of human and bovine milk.

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    Kasper Hettinga

    Full Text Available Milk is the single source of nutrients for the newborn mammal. The composition of milk of different mammals has been adapted during evolution of the species to fulfill the needs of the offspring. Milk not only provides nutrients, but it also serves as a medium for transfer of host defense components to the offspring. The host defense proteins in the milk of different mammalian species are expected to reveal signatures of evolution. The aim of this study is therefore to study the difference in the host defense proteome of human and bovine milk. We analyzed human and bovine milk using a shot-gun proteomics approach focusing on host defense-related proteins. In total, 268 proteins in human milk and 269 proteins in bovine milk were identified. Of these, 44 from human milk and 51 from bovine milk are related to the host defense system. Of these proteins, 33 were found in both species but with significantly different quantities. High concentrations of proteins involved in the mucosal immune system, immunoglobulin A, CD14, lactoferrin, and lysozyme, were present in human milk. The human newborn is known to be deficient for at least two of these proteins (immunoglobulin A and CD14. On the other hand, antimicrobial proteins (5 cathelicidins and lactoperoxidase were abundant in bovine milk. The high concentration of lactoperoxidase is probably linked to the high amount of thiocyanate in the plant-based diet of cows. This first detailed analysis of host defense proteins in human and bovine milk is an important step in understanding the function of milk in the development of the immune system of these two mammals.

  13. Quantitative milk genomics: estimation of variance components and prediction of fatty acids in bovine milk

    DEFF Research Database (Denmark)

    Krag, Kristian

    The composition of bovine milk fat, used for human consumption, is far from the recommendations for human fat nutrition. The aim of this PhD was to describe the variance components and prediction probabilities of individual fatty acids (FA) in bovine milk, and to evaluate the possibilities...

  14. Structural and functional insights into the catalytic inactivity of the major fraction of buffalo milk xanthine oxidoreductase.

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    Kaustubh S Gadave

    Full Text Available BACKGROUND: Xanthine oxidoreductase (XOR existing in two interconvertible forms, xanthine dehydrogenase (XDH and xanthine oxidase (XO, catabolises xanthine to uric acid that is further broken down to antioxidative agent allantoin. XOR also produces free radicals serving as second messenger and microbicidal agent. Large variation in the XO activity has been observed among various species. Both hypo and hyper activity of XOR leads to pathophysiological conditions. Given the important nutritional role of buffalo milk in human health especially in south Asia, it is crucial to understand the functional properties of buffalo XOR and the underlying structural basis of variations in comparison to other species. METHODS AND FINDINGS: Buffalo XO activity of 0.75 U/mg was almost half of cattle XO activity. Enzymatic efficiency (k cat/K m of 0.11 sec(-1 µM(-1 of buffalo XO was 8-10 times smaller than that of cattle XO. Buffalo XOR also showed lower antibacterial activity than cattle XOR. A CD value (Δε430 nm of 46,000 M(-1 cm(-1 suggested occupancy of 77.4% at Fe/S I centre. Buffalo XOR contained 0.31 molybdenum atom/subunit of which 48% existed in active sulfo form. The active form of XO in buffalo was only 16% in comparison to ∼30% in cattle. Sequencing revealed 97.4% similarity between buffalo and cattle XOR. FAD domain was least conserved, while metal binding domains (Fe/S and Molybdenum were highly conserved. Homology modelling of buffalo XOR showed several variations occurring in clusters, especially close to FAD binding pocket which could affect NAD(+ entry in the FAD centre. The difference in XO activity seems to be originating from cofactor deficiency, especially molybdenum. CONCLUSION: A major fraction of buffalo milk XOR exists in a catalytically inactive form due to high content of demolybdo and desulfo forms. Lower Fe/S content and structural factors might be contributing to lower enzymatic efficiency of buffalo XOR in a minor way.

  15. Genes involved in bovine milk-fat composition

    NARCIS (Netherlands)

    Schennink, A.

    2009-01-01

    The aim of the research described in this thesis was to identify genes that underlie the genetic variation in bovine milk-fat composition. The fat composition of milk samples from approximately 2,000 Dutch Holstein Friesian cows in their first lactation was measured by gas chromatography. Quantita

  16. Bovine milk usage and feeding practices for infants in India

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    Meha Mayuri

    2012-01-01

    Full Text Available Background: Milk and dairy products from animal sources (cow′s milk or buffalo′s milk are an important part of Indian diet and its consumption starts at an early age. Objective: The purpose of this study is to understand the processing, storage and modifications carried out before bovine milk is fed to the infants and compare it with recommendations by Health Care Professionals (HCPs. Materials and Methods: The study involved assessments involving HCPs and mothers of children below 1 year of age through both qualitative and quantitative methods. Feeding recommendations by HCPs were assessed through a quantitative method. Results: The children are commonly initiated on bovine milk post 3 months of age. The milk is not covered while boiling in 70% cases and boiled at suboptimal temperature (moderate: 62% or low: 31%. Half of the families store milk outside refrigerators openly in the kitchen or rooms. The milk is modified in the majority of cases (81% before being fed. Modification of the milk was often done with sugar (85%, followed by water (49%, biscuits, fennel seeds, cardamom or infant cereals before feeding the baby. Addition of water was more prevalent among mothers of infants between 3 and 6 months. HCPs do not advise consumption of bovine milk by infants unless there is no other option available. Conclusion: A clear gap exists between recommended and actual practices for infant feeding in India. There is a huge opportunity to educate mothers on importance of breast feeding and benefits of implementing appropriate processing, storage, and consumption practices of bovine milk.

  17. Characterization of carbohydrate structures of bovine MUC15 and distribution of the mucin in bovine milk

    DEFF Research Database (Denmark)

    Pallesen, Lone Tjener; Pedersen, Lise Refstrup Linnebjerg; Petersen, Torben Ellebæk

    2007-01-01

    -containing fractions as well, such as skim milk and whey. Compositional and structural studies of the carbohydrates of bovine milk MUC15 showed that the glycans are composed of fucose, galactose, mannose, N-acetylgalactosamine, N-acetylglycosamine, and sialic acid. The carbohydrate was shown to constitute 65......The present work reports the characterization of carbohydrate structures and the distribution of the newly identified mucin MUC15, a highly glycosylated protein associated with the bovine milk fat globule membrane (MFGM). Distribution of MUC15 was investigated in various fractions of bovine milk...... by densitometric scanning of Western blots. In raw milk, MUC15 was shown to constitute 0.08% (wt) of the protein and approximately 1.5% (wt) of the MFGM-associated proteins. Surprisingly, this study showed that in addition to the fat-containing fractions, such as MFGM and buttermilk, MUC15 was present in nonfat...

  18. Metabolomic Approaches to Explore Chemical Diversity of Human Breast-Milk, Formula Milk and Bovine Milk.

    Science.gov (United States)

    Qian, Linxi; Zhao, Aihua; Zhang, Yinan; Chen, Tianlu; Zeisel, Steven H; Jia, Wei; Cai, Wei

    2016-12-17

    Although many studies have been conducted on the components present in human breast milk (HM), research on the differences of chemical metabolites between HM, bovine milk (BM) and formula milk (FM) is limited. This study was to explore the chemical diversity of HM, BM and FM by metabolomic approaches. GC-TOFMS and UPLC-QTOFMS were applied to investigate the metabolic compositions in 30 HM samples, 20 FM samples and 20 BM samples. Metabolite profiling identified that most of the non-esterified fatty acids, which reflected the hydrolysis of triglycerides, were much more abundant in HM than those in FM and BM, except for palmitic acid and stearic acid. The levels of tricarboxylic acid (TCA) intermediates were much higher in FM and BM than those in HM. Each type of milk also showed its unique composition of free amino acids and free carbohydrates. In conclusion, higher levels of non-esterified saturated fatty acids with aliphatic tails <16 carbons, monounsaturated fatty acids and polyunsaturated fatty acids and lower levels of TCA intermediates are characteristic of HM, as compared with FM and BM. The content of non-esterified fatty acids may reflect the hydrolysis of triglycerides in different milk types.

  19. Fluorometric determination of free and total isocitrate in bovine milk

    DEFF Research Database (Denmark)

    Larsen, Torben

    2014-01-01

    and cholesterol synthesis. This study describes a new analytical method for determination of free and total isocitrate in bovine milk where time-consuming pretreatment of the sample is not necessary. Methods for estimation of both total isocitrate and free isocitrate are described, the difference being....... Milk samples from Danish Holstein cows (984) contained significantly less isocitrate than milk from Danish Jersey cows (760; i.e., 0.134 vs. 0.211 mmol/L). Isocitrate in milk is correlated to milk protein, fat, and citrate, and it is speculated, based on biochemistry, former studies, and the present...... the esterified or even lactonized isocitrate. On average, 20% (6–27%) of cow milk isocitrate was esterified and free isocitrate correlated significantly with total isocitrate (r = 0.98). The present fluorometric determination correlated well with the traditional spectrophotometric determination of isocitrate...

  20. Expanding the bovine milk proteome through extensive fractionation

    DEFF Research Database (Denmark)

    Nissen, Asger; Bendixen, Emøke; Ingvartsen, Klaus Lønne

    2013-01-01

    sensitivity limit of the 2-dimensional liquid chromatography tandem mass spectrometry analysis. Each fractionation technique resulted in identification of a unique subset of proteins. However, high-speed centrifugation of milk to whey was by far the best method to achieve high and repeatable proteome coverage......Bovine milk is an agricultural product of tremendous value worldwide. It contains proteins, fat, lactose, vitamins, and minerals. It provides nutrition and immunological protection (e.g., in the gastrointestinal tract) to the newborn and young calf. It also forms an important part of human...... nutrition. The repertoire of proteins in milk (i.e., its proteome) is vast and complex. The milk proteome can be described in detail by mass spectrometry-based proteomics. However, the high concentration of dominating proteins in milk reduces mass spectrometry detection sensitivity and limits detection...

  1. Genetic variation in bovine milk fat composition

    NARCIS (Netherlands)

    Stoop, W.M.

    2009-01-01

    In her thesis, Stoop shows that there is considerable genetic variation in milk fat composition, which opens opportunities to improve milk fat composition by selective breeding. Short and medium chain fatty acids had high heritabilities, whereas variation due to herd (mainly feed effects) was modera

  2. Antibody-independent identification of bovine milk-derived peptides in breast-milk.

    Science.gov (United States)

    Picariello, Gianluca; Addeo, Francesco; Ferranti, Pasquale; Nocerino, Rita; Paparo, Lorella; Passariello, Annalisa; Dallas, David C; Robinson, Randall C; Barile, Daniela; Canani, Roberto Berni

    2016-08-10

    Exclusively breast-fed infants can exhibit clear signs of IgE or non IgE-mediated cow's milk allergy. However, the definite characterization of dietary cow's milk proteins (CMP) that survive the maternal digestive tract to be absorbed into the bloodstream and secreted into breast milk remains missing. Herein, we aimed at assessing possible CMP-derived peptides in breast milk. Using high performance liquid chromatography (HPLC)-high resolution mass spectrometry (MS), we compared the peptide fraction of breast milk from 12 donors, among which 6 drank a cup of milk daily and 6 were on a strict dairy-free diet. We identified two bovine β-lactoglobulin (β-Lg, 2 out 6 samples) and one αs1-casein (1 out 6 samples) fragments in breast milk from mothers receiving a cup of bovine milk daily. These CMP-derived fragments, namely β-Lg (f42-54), (f42-57) and αs1-casein (f180-197), were absent in milk from mothers on dairy-free diet. In contrast, neither intact nor hydrolyzed β-Lg was detected by western blot and competitive ELISA in any breast milk sample. Eight additional bovine milk-derived peptides identified by software-assisted MS were most likely false positive. The results of this study demonstrate that CMP-derived peptides rather than intact CMP may sensitize or elicit allergic responses in the neonate through mother's milk. Immunologically active peptides from the maternal diet could be involved in priming the newborn's immune system, driving a tolerogenic response.

  3. Transcriptional profiling of bovine milk using RNA sequencing

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    Wickramasinghe Saumya

    2012-01-01

    Full Text Available Abstract Background Cow milk is a complex bioactive fluid consumed by humans beyond infancy. Even though the chemical and physical properties of cow milk are well characterized, very limited research has been done on characterizing the milk transcriptome. This study performs a comprehensive expression profiling of genes expressed in milk somatic cells of transition (day 15, peak (day 90 and late (day 250 lactation Holstein cows by RNA sequencing. Milk samples were collected from Holstein cows at 15, 90 and 250 days of lactation, and RNA was extracted from the pelleted milk cells. Gene expression analysis was conducted by Illumina RNA sequencing. Sequence reads were assembled and analyzed in CLC Genomics Workbench. Gene Ontology (GO and pathway analysis were performed using the Blast2GO program and GeneGo application of MetaCore program. Results A total of 16,892 genes were expressed in transition lactation, 19,094 genes were expressed in peak lactation and 18,070 genes were expressed in late lactation. Regardless of the lactation stage approximately 9,000 genes showed ubiquitous expression. Genes encoding caseins, whey proteins and enzymes in lactose synthesis pathway showed higher expression in early lactation. The majority of genes in the fat metabolism pathway had high expression in transition and peak lactation milk. Most of the genes encoding for endogenous proteases and enzymes in ubiquitin-proteasome pathway showed higher expression along the course of lactation. Conclusions This is the first study to describe the comprehensive bovine milk transcriptome in Holstein cows. The results revealed that 69% of NCBI Btau 4.0 annotated genes are expressed in bovine milk somatic cells. Most of the genes were ubiquitously expressed in all three stages of lactation. However, a fraction of the milk transcriptome has genes devoted to specific functions unique to the lactation stage. This indicates the ability of milk somatic cells to adapt to different

  4. Expanding the bovine milk proteome through extensive fractionation.

    Science.gov (United States)

    Nissen, Asger; Bendixen, Emøke; Ingvartsen, Klaus Lønne; Røntved, Christine Maria

    2013-01-01

    Bovine milk is an agricultural product of tremendous value worldwide. It contains proteins, fat, lactose, vitamins, and minerals. It provides nutrition and immunological protection (e.g., in the gastrointestinal tract) to the newborn and young calf. It also forms an important part of human nutrition. The repertoire of proteins in milk (i.e., its proteome) is vast and complex. The milk proteome can be described in detail by mass spectrometry-based proteomics. However, the high concentration of dominating proteins in milk reduces mass spectrometry detection sensitivity and limits detection of low abundant proteins. Further, the general health and udder health of the dairy cows delivering the milk may influence the composition of the milk proteome. To gain a more exhaustive and true picture of the milk proteome, we performed an extensive preanalysis fractionation of raw composite milk collected from documented healthy cows in early lactation. Four simple and industrially applicable techniques exploring the physical and chemical properties of milk, including acidification, filtration, and centrifugation, were used for separation of the proteins. This resulted in 5 different fractions, whose content of proteins were compared with the proteins of nonfractionated milk using 2-dimensional liquid chromatography tandem mass spectrometry analysis. To validate the proteome analysis, spectral counts and ELISA were performed on 7 proteins using the ELISA for estimation of the detection sensitivity limit of the 2-dimensional liquid chromatography tandem mass spectrometry analysis. Each fractionation technique resulted in identification of a unique subset of proteins. However, high-speed centrifugation of milk to whey was by far the best method to achieve high and repeatable proteome coverage. The total number of milk proteins initially detected in nonfractionated milk and the fractions were 635 in 2 replicates. Removal of dominant proteins and filtering for redundancy across the

  5. Short communication: Milk ELISA status for bovine leukosis virus infection is not associated with milk production in dairy cows.

    Science.gov (United States)

    Sorge, U S; Lissemore, K; Cantin, R; Kelton, D F

    2011-10-01

    The objective of this study was to assess whether the milk ELISA status for antibodies against bovine leukemia virus was associated with 305-d milk production in Canadian dairy cattle. Test results and test-day production data from 19,785 dairy cows were available for analysis. A linear mixed model was used with the estimated 305-d milk production as the outcome and lactation number, somatic cell count, calving season, days in milk, and breed as fixed effects. Herd nested in province was included as random effect. In conclusion, bovine leukemia virus antibody milk ELISA status was not associated with milk production.

  6. Mycotoxins in Bovine Milk and Dairy Products: A Review.

    Science.gov (United States)

    Becker-Algeri, Tania Aparecida; Castagnaro, Denise; de Bortoli, Kennidy; de Souza, Camila; Drunkler, Deisy Alessandra; Badiale-Furlong, Eliana

    2016-03-01

    This paper presents a literature review of the occurrence of several mycotoxins in bovine milk and dairy products, because it is the main type of milk produced and marketed worldwide. Mycotoxins are produced by different genera of filamentous fungi and present serious health hazards such as carcinogenicity and mutagenicity. Under favorable growth conditions, toxigenic fungi produce mycotoxins which contaminate the lactating cow's feedstuff. During metabolism, these mycotoxins undergo biotransformation and are secreted in milk. Data show that there is a seasonal trend in the levels of mycotoxins in milk, with these being higher in the cold months probably due to the prolonged storage required for the cattle feeds providing favorable conditions for fungal growth. Good agricultural and storage practices are therefore of fundamental importance in the control of toxigenic species and mycotoxins. Although aflatoxins (especially aflatoxin M1 ) are the mycotoxins of greater incidence in milk and dairy products, this review shows that other mycotoxins, such as fumonisin, ochratoxin A, trichothecenes, zearalenone, T-2 toxin, and deoxynivalenol, can also be found in these products. Given that milk is widely consumed and is a source of nutrients, especially in childhood, a thorough investigation of the occurrence of mycotoxins as well the adoption of measures to minimize their contamination of milk is essential.

  7. Milk adulteration: Detection of bovine milk in bulk goat milk produced by smallholders in northeastern Brazil by a duplex PCR assay.

    Science.gov (United States)

    Rodrigues, N P A; Givisiez, P E N; Queiroga, R C R E; Azevedo, P S; Gebreyes, W A; Oliveira, C J B

    2012-05-01

    The aim of this study was to investigate the adulteration of goat milk produced by smallholders in semiarid northeastern Brazil with bovine milk as an adulterant. The study was requested by the association of smallholder producers in the region to investigate and to inhibit adulteration practices as a need to ensure the quality and safety of goat milk. A duplex PCR assay has been developed and standardized. Further validation was performed in 160 fresh bulk goat milk samples. The detection limit of the duplex PCR was 0.5% bovine milk in goat milk and the results indicated that 41.2% of the goat milk presented to market was positive for bovine milk. Making the test available to the association of producers, together with extension activities, have been applied to reduce adulteration in goat milk sold to small-scale dairy plants and to ensure the species origin for goat milk in the state of Paraíba.

  8. Immunochromatographic Lateral-flow test strip for the rapid detection of added bovine rennet whey in milk and milk powder

    NARCIS (Netherlands)

    Martin-Hernandez, C.; Munoz, M.; Daury, C.; Weymuth, H.; Kemmers-Voncken, A.; Corbation, V.; Toribo, T.; Bremer, M.G.E.G.

    2009-01-01

    An immunochromatographic lateral-flow test dipstick test was developed for the fast detection of bovine rennet whey in liquid milk and milk powder. The test is based on the binding of casein glycomacropeptide (cGMP) by two specific anti-bovine ¿-casein monoclonal antibodies and has a visual detectio

  9. Immunoglobulin A in Bovine Milk: A Potential Functional Food?

    Science.gov (United States)

    Cakebread, Julie A; Humphrey, Rex; Hodgkinson, Alison J

    2015-08-26

    Immunoglobulin A (IgA) is an anti-inflammatory antibody that plays a critical role in mucosal immunity. It is found in large quantities in human milk, but there are lower amounts in bovine milk. In humans, IgA plays a significant role in providing protection from environmental pathogens at mucosal surfaces and is a key component for the establishment and maintenance of intestinal homeostasis via innate and adaptive immune mechanisms. To date, many of the dairy-based functional foods are derived from bovine colostrum, targeting the benefits of IgG. IgA has a higher pathogenic binding capacity and greater stability against proteolytic degradation when ingested compared with IgG. This provides IgA-based products greater potential in the functional food market that has yet to be realized.

  10. Biosensor assay for determination of haptoglobin in bovine milk.

    Science.gov (United States)

    Akerstedt, Maria; Björck, Lennart; Persson Waller, Karin; Sternesjö, Ase

    2006-08-01

    Despite more than 30 years of research into mastitis diagnostics, there are few alternatives to the somatic cell count (SCC) in practical use for identification of cows with subclinical mastitis. Mastitis is not only an animal welfare problem, but also affects the yield, composition and technological properties of milk. Hence, dairy cooperatives give farmers a premium quality payment to encourage low SCC although there is no clear scientific data defining the level of SCC in bulk tank milk that is associated with additional benefits in terms of milk quality. Recent research on alternative markers for inflammatory reactions in the lactating cow, e.g. in mastitis, includes investigations of the acute phase protein, haptoglobin (Hp). So far, the content of Hp in milk has mainly been studied in relation to mastitis diagnostics, with little attention given to its importance for milk composition and technological properties. At present, Hp in milk is measured using ELISA, but this technique is not suitable for routine large-scale analysis. In recent years, optical biosensor technology has been used for automated and rapid quantitative analysis of different components in milk, but so far not for analysis of acute phase proteins. The aim of the present study was to develop a rapid and sensitive biosensor method to determine Hp in milk. An affinity sensor assay based on the interaction between Hp and haemoglobin was developed using surface plasmon resonance (SPR) biosensor technology. The assay was used to analyse Hp in composite milk samples from cows without any clinical signs of mastitis and quarter milk samples with a weak to strong reaction in the California Mastitis Test (CMT). A commercial ELISA for determination of Hp in milk was used for comparison. The limit of detection (LOD) of the biosensor assay was determined as 1.1 mg/l. Within-assay and between-day variations were determined both with bulk tank milk spiked with human Hp and with composite milk samples

  11. Separation of plasminogen activator in bovine milk, and its various days after delivery

    OpenAIRE

    堀江, 登

    2000-01-01

    Plasminogen activator was separated by gel-filtration of skimmed milk from bovine until 9^ day after delivery. The separation pattern of plasminogen activator in bovine milk was different from one of human milk. The activities of plasminogen activator in bovine milk were compared among three groups of skimmed milk samples obtained in 1~3day, 4~6day, 7~9day after delivery. The mean value for each group were 3.2IU/ml(l~3day), 3.5IU/ml(4~6day) and 0.3IU/ml(4~9day).

  12. Detecting β-Casein Variation in Bovine Milk

    Directory of Open Access Journals (Sweden)

    Anna Maria Caroli

    2016-01-01

    Full Text Available In bovine species, β-casein (β-CN is characterized by genetic polymorphism. The two most common protein variants are β-CN A2 (the original one and A1, differing from A2 for one amino acid substitution (Pro67 to His67. Several bioactive peptides affecting milk nutritional properties can originate from β-CN. Among them, β-casomorphin-7 (BCM7 ranging from amino acid 60 to 66 can be released more easily from β-CN variants carrying His67 (A1 type instead of Pro67 (A2 type. Nowadays, “A2 milk” is produced in different countries claiming its potential benefits in human health. The aim of this study was to further develop and apply an isoelectric focusing electrophoresis (IEF method to bulk and individual milk samples in order to improve its use for β-CN studies. We succeeded in identifying A2 milk samples correctly and quantifying the percentage of A2, A1, and B variants in bulk samples not derived from A2 milk as well as in individual milk samples. The method allows us to quantify the relative proportion of β-CN variants in whole milk without eliminating whey protein by acid or enzymatic precipitation of caseins. The aim of this study was also to study the different behavior of β-CN and β-lactoglobulin (β-LG in the presence of trichloroacetic acid (TCA. The higher sensitivity of β-CN to TCA allows quantifying β-CN variants after TCA fixation because β-LG is not visible. Monitoring β-CN variation in cattle breeds is important in order to maintain a certain balance between Pro67 and His67 in dairy products. Overall, the debate between A1 and A2 milk needs further investigation.

  13. Modification of the Kjeldahl noncasein nitrogen method to include bovine milk concentrates and milks from other species.

    Science.gov (United States)

    Wojciechowski, Karen L; Barbano, David M

    2015-11-01

    The objective of our research was to modify the current indirect casein method for bovine milk to enable it to be applied to bovine milk, bovine milk concentrates, and milks of other species that contain a protein concentration up to 9% (wt/wt). Our work used a series of bovine milk concentrates from about 3 to 9% protein with the same casein as a percentage of true protein to determine the amount of buffer required and pH of the noncasein nitrogen (NCN) filtrate to achieve consistent estimates of casein and casein as percent of true protein. As the concentration of protein in milk increased (either in bovine milk concentrates or in milks of other species), the amount of buffer needed for the NCN sample preparation method to achieve a filtrate pH of 4.6 increased. In the first part of the study using a series of bovine milk concentrates, it was demonstrated that the method gave more consistent predictions of casein as a percentage of true protein when the final NCN filtrate pH was between 4.5 and 4.6 at 38°C. When the amount of buffer added to the sample was not sufficient (i.e., the filtrate pH was too high), the filtrates were not clear. A polynomial equation was developed for prediction of the amount of acetic acid or sodium acetate buffer required to achieve pH 4.5 to 4.6 for milk protein concentrations from 3 to 9% protein using bovine milk and milk concentrates. When the equation developed using cow milk was applied to goat, sheep, and water buffalo milks, it correctly predicted the volume of reagents needed to achieve a final NCN filtrate pH of 4.6 at 38°C. We also verified as part of this work that the ability to measure NPN content of milk was not influenced by protein content of milk in the range from 3 to 9% protein. The results of this study will be used as the basis for proposed changes in the official methods for measurement of the casein content of milk to expand the scope of the method so it can be used to achieve accurate results for milk

  14. Xanthine urolithiasis

    Directory of Open Access Journals (Sweden)

    Gargah Tahar

    2010-01-01

    Full Text Available Hereditary xanthinuria type I, a defect of purine matabolism, results from a genetic deficiency of xanthine oxidase. It is an uncommon cause of stone formation in children. We report here two children with xanthine urolithiasis. The first patient was an 8-year-old boy who presented with repeated episodes of hematuria evaluated with excretory urography, which demonstrated radio-lucent pelvic stone in the right kidney, causing hydronephrosis. He had pyelolithotomy, and the extracted stone consisted of pure xanthine. Family study revealed an asymptomatic xanthinuria in younger brother. The second patient was a 5-year-old boy who had a 2-week history of abdominal pain and gross hematuria. Conventional excretory intravenous urography showed a non-functioning right kidney. Nephrectomy was performed, and histology revealed end-stage pyelonephritis. The calculi consisted of pure xanthine. In both patients, plasma and urinary concentrations of uric acid were low but xanthine and hypoxanthine concentrations were markedly elevated. Xanthine uroli-thiasis is usually a benign condition, easy to prevent or cure by appropriate alkalinization, forced hydration and restriction of dietary purines. However asymptomatic, and therefore undiagnosed, stones may invade the kidney and urinary tract, resulting in destruction of parenchyma, nephrec-tomy and renal failure.

  15. Genetic variability of fatty acids in bovine milk

    Directory of Open Access Journals (Sweden)

    Soyeurt H.

    2008-01-01

    Full Text Available Fatty acids composition of bovine milk influences the technological properties of butterfat and also presents some potential benefits for human health. Impact of feeding on fat composition is well described in the literature; less information is available about the impact of genetics. Based on few studies, essentially conducted to isolate some feeding effect, the breed seemed to influence the fatty acids composition. The variation in the activity of δ-9 desaturase, key enzyme in the production of monounsaturated fatty acids and conjugated linoleic acids in milk, could explain these differences. Very few studies have been focussing on the estimation of genetic parameters of fatty acids composition. However, the moderate heritability estimates observed by these studies for the major fatty acids could suggest a potential genetic effect.

  16. Analysis of fenbendazole residues in bovine milk by ELISA.

    Science.gov (United States)

    Brandon, David L; Bates, Anne H; Binder, Ronald G; Montague, William C; Whitehand, Linda C; Barker, Steven A

    2002-10-09

    Fenbendazole residues in bovine milk were analyzed by ELISAs using two monoclonal antibodies. One monoclonal antibody (MAb 587) bound the major benzimidazole anthelmintic drugs, including fenbendazole, oxfendazole, and fenbendazole sulfone. The other (MAb 591) was more specific for fenbendazole, with 13% cross-reactivity with the sulfone and no significant binding to the sulfoxide metabolite. The limit of detection of the ELISA method in the milk matrix was 7 ppb for MAb 587 and 3 ppb for MAb 591. Fenbendazole was administered in feed, drench, and paste form to three groups of dairy cattle. Milk was collected immediately before dosing and then every 12 h for 5 days. The ELISA indicated that residue levels varied widely among individual cows in each group. Fenbendazole levels peaked at approximately 12-24 h and declined rapidly thereafter. Metabolites were detected at much higher levels than the parent compound, peaked at approximately 24-36 h, and declined gradually. Residue levels were undetectable by 72 h. The ELISA data correlated well with the total residues determined by chromatographic analysis, but the use of the two separate ELISAs did not afford an advantage over ELISA with the single, broadly reactive MAb 587. The ELISA method could be used to flag high-residue samples in on-site monitoring of fenbendazole in milk and is a potential tool for studying drug pharmacokinetics.

  17. Detection of bovine herpesvirus 4 glycoprotein B and thymidine kinase DNA by PCR assays in bovine milk

    NARCIS (Netherlands)

    Wellenberg, G.J.; Verstraten, E.; Belak, S.; Verschuren, S.B.E.; Rijsewijk, F.A.M.; Peshev, R.; Oirschot, van J.T.

    2001-01-01

    A polymerase chain reaction (PCR) assay was developed to detect bovine herpesvirus 4 (BHV4) glycoprotein B (gB) DNA, and a nested-PCR assay was modified for the detection of BHV4 thymidine kinase (TK) DNA in bovine milk samples. To identify false-negative PCR results, internal control templates were

  18. Inhibitory activity of xanthine oxidase by fractions Crateva adansonii

    Institute of Scientific and Technical Information of China (English)

    Abdullahi A; Kolo MZ; Hamzah RU; Jigam AA; Yahya A; Kabiru AY; Muhammad H; Sakpe S; Adefolalu FS; Isah MC

    2012-01-01

    Objective: To study the inhibitory effect of various extracts from Crateva adansonii (C. adansonii) used traditionally against several inflammatory diseases such as rheumatism, arthritis, and gout, was investigated on purified bovine milk xanthine oxidase (XO) activity. Methods:Xanthine oxidase inhibitory activity was assayed spectrophotometrically and the degree of enzyme inhibition was determined by measuring the increase in absorbance at 295 nm associated with uric acid formation. Enzyme kinetics was carried out using Lineweaver-Burk plots using xanthine as the substrate. Results: Among the fractions tested, the chloroform fraction exhibited highest potency (IC50 20.2±1.6 μg/mL) followed by the petroleum ether (IC50 30.1±2.2 μg/mL), ethyl acetate (IC50 43.9±1.4 μg/mL) and residual (IC50 98.0±3.3 μg/mL) fractions. The IC50 value of allopurinol used, as the standard was 5.7±0.3 μg/mL. Conclusions: Enzyme inhibition mechanism indicated that the mode of inhibition was of a mixed type. Our findings suggest that the therapeutic use of these plants may be due to the observed Xanthine oxidase inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.

  19. Effect of bovine lactoferrin addition to milk in yogurt manufacturing.

    Science.gov (United States)

    Franco, I; Castillo, E; Pérez, M D; Calvo, M; Sánchez, L

    2010-10-01

    The aim of this work was to study the effect of milk supplementation with lactoferrin of different iron saturation on the manufacturing and characteristics of yogurt. Bovine lactoferrin was added at concentrations of 0.5, 1, and 2 mg/mL in the holo (iron saturated) and apo (without iron) forms. Some physicochemical properties, such as pH, concentration of lactic acid, and texture of supplemented yogurts, were determined throughout the shelf-life period storage (28 d) at 4°C. We also evaluated the stability of lactoferrin in supplemented yogurt throughout the storage time. The supplementation of milk with bovine lactoferrin did not greatly affect the physical properties of the yogurt, though apo-lactoferrin slightly delayed the decrease of pH. This could be attributed to the partial inhibition observed on the growth of Streptococcus thermophilus. The integrity and immunoreactive concentration of lactoferrin, determined by Western blotting and noncompetitive ELISA, respectively, remained constant throughout the shelf life of yogurt.

  20. STAT6-dependent collagen synthesis in human fibroblasts is induced by bovine milk

    OpenAIRE

    Stefan Kippenberger; Nadja Zöller; Johannes Kleemann; Jutta Müller; Roland Kaufmann; Matthias Hofmann; August Bernd; Markus Meissner; Eva Valesky

    2015-01-01

    Since the domestication of the urus, 10.000 years ago, mankind utilizes bovine milk for different purposes. Besides usage as a nutrient also the external application of milk on skin has a long tradition going back to at least the ancient Aegypt with Cleopatra VII as a great exponent. In order to test whether milk has impact on skin physiology, cultures of human skin fibroblasts were exposed to commercial bovine milk. Our data show significant induction of proliferation by milk (max. 2,3-fold,...

  1. Bovine milk-derived exosomes for drug delivery.

    Science.gov (United States)

    Munagala, Radha; Aqil, Farrukh; Jeyabalan, Jeyaprakash; Gupta, Ramesh C

    2016-02-01

    Exosomes are biological nanovesicles that are involved in cell-cell communication via the functionally-active cargo (such as miRNA, mRNA, DNA and proteins). Because of their nanosize, exosomes are explored as nanodevices for the development of new therapeutic applications. However, bulk, safe and cost-effective production of exosomes is not available. Here, we show that bovine milk can serve as a scalable source of exosomes that can act as a carrier for chemotherapeutic/chemopreventive agents. Drug-loaded exosomes showed significantly higher efficacy compared to free drug in cell culture studies and against lung tumor xenografts in vivo. Moreover, tumor targeting ligands such as folate increased cancer-cell targeting of the exosomes resulting in enhanced tumor reduction. Milk exosomes exhibited cross-species tolerance with no adverse immune and inflammatory response. Thus, we show the versatility of milk exosomes with respect to the cargo it can carry and ability to achieve tumor targetability. This is the first report to identify a biocompatible and cost-effective means of exosomes to enhance oral bioavailability, improve efficacy and safety of drugs.

  2. Detection and characterisation of Complement protein activity in bovine milk by bactericidal sequestration assay.

    Science.gov (United States)

    Maye, Susan; Stanton, Catherine; Fitzgerald, Gerald F; Kelly, Philip M

    2015-08-01

    While the Complement protein system in human milk is well characterised, there is little information on its presence and activity in bovine milk. Complement forms part of the innate immune system, hence the importance of its contribution during milk ingestion to the overall defences of the neonate. A bactericidal sequestration assay, featuring a Complement sensitive strain, Escherichia coli 0111, originally used to characterise Complement activity in human milk was successfully applied to freshly drawn bovine milk samples, thus, providing an opportunity to compare Complement activities in both human and bovine milks. Although not identical in response, the levels of Complement activity in bovine milk were found to be closely comparable with that of human milk. Differential counts of Esch. coli 0111 after 2 h incubation were 6.20 and 6.06 log CFU/ml, for raw bovine and human milks, respectively - the lower value representing a stronger Complement response. Exposing bovine milk to a range of thermal treatments e.g. 42, 45, 65, 72, 85 or 95 °C for 10 min, progressively inhibited Complement activity by increasing temperature, thus confirming the heat labile nature of this immune protein system. Low level Complement activity was found, however, in 65 and 72 °C heat treated samples and in retailed pasteurised milk which highlights the outer limit to which high temperature, short time (HTST) industrial thermal processes should be applied if retention of activity is a priority. Concentration of Complement in the fat phase was evident following cream separation, and this was also reflected in the further loss of activity recorded in low fat variants of retailed pasteurised milk. Laboratory-based churning of the cream during simulated buttermaking generated an aqueous (buttermilk) phase with higher levels of Complement activity than the fat phase, thus pointing to a likely association with the milk fat globule membrane (MFGM) layer.

  3. A novel isolation strategy for obtaining crude membrane vesicles from bovine skim milk

    DEFF Research Database (Denmark)

    Blans, Kristine; Larsen, Lotte Bach; Wiking, Lars

    2014-01-01

    Bovine milks content of phospholipid membranes have largely been explored in the cream fraction, and known as the milk fat globule membrane that surrounds fat droplets. In skim milk, the population of phospholipid membranes is reported to constitute membrane vesicles with a soluble content known...... components. Here we present a novel strategy for a short, gentle and non-denaturing isolation of skim milk-derived membrane vesicles. Methods: Untreated fresh bovine milk was defatted to remove milk fat globules. The resulting skim milk was subjected to ultracentrifugation. The resulting ochre...... fraction can be obtained from skim milk by ultracentrifugation. Casein micelle remnants as well as smaller protein components in the crude vesicle fraction can be successfully removed by size chromatography. Electron microscopy of the vesicle isolate reveals circular structures with membrane vesicle...

  4. A novel isolation strategy for obtaining crude membrane vesicles from bovine skim milk

    DEFF Research Database (Denmark)

    Blans, Kristine; Larsen, Lotte Bach; Wiking, Lars

    Bovine milks content of phospholipid membranes have largely been explored in the cream fraction, and known as the milk fat globule membrane that surrounds fat droplets. In skim milk, the population of phospholipid membranes is reported to constitute membrane vesicles with a soluble content known...... components. Here we present a novel strategy for a short, gentle and non-denaturing isolation of skim milk-derived membrane vesicles. Methods: Untreated fresh bovine milk was defatted to remove milk fat globules. The resulting skim milk was subjected to ultracentrifugation. The resulting ochre...... fraction can be obtained from skim milk by ultracentrifugation. Casein micelle remnants as well as smaller protein components in the crude vesicle fraction can be successfully removed by size chromatography. Electron microscopy of the vesicle isolate reveals circular structures with membrane vesicle...

  5. Effect of Technological Treatments on Human-Like Leptin Level in Bovine Milk for Human Consumption

    Directory of Open Access Journals (Sweden)

    Damiano Magistrelli

    2014-07-01

    Full Text Available In this experiment, raw milk and commercially available full-cream UHT milk, semi-skimmed UHT milk, skimmed UHT milk, full-cream pasteurized milk, semi-skimmed pasteurized milk and infant formulas for babies between 6 and 12 months of age were analyzed by RIA, with a method using an antibody directed against human leptin and human leptin as reference standard. Raw milk and full-cream UHT milk did not differ for human-like leptin. Leptin content of full-cream pasteurized milk was not different to that of full-cream UHT milk, but it was 14% lower (p < 0.05 than that observed in raw milk. Human-like leptin level of semi-skimmed UHT milk was not different to that of semi-skimmed pasteurized milk, but it was 30% lower (p < 0.0001 than those of full-cream UHT and full-cream pasteurized milks. In skimmed UHT milk, leptin was 40% lower (p < 0.0001 than in full-cream UHT milk. Leptin was correlated (p < 0.001 with lipid content. Leptin level of infant formulas was not different to that of skimmed milks. Results suggest that the heat treatment (pasteurization or UHT is not a modifier of human-like leptin content of edible commercial bovine milks, whereas the skimming process significantly reduces milk leptin level.

  6. Bovine milk osteopontin - Targeting bacterial adhesion for biofilm control

    DEFF Research Database (Denmark)

    Kristensen, Mathilde Frost; Meyer, Rikke Louise; Schlafer, Sebastian

    2016-01-01

    Self-performed mechanical tooth cleaning does usually not result in complete biofilm removal, due to the complex oral anatomy and the strong adhesion of the biofilm to the tooth. Therefore, different supportive measures are employed, most of which aim at the chemical eradication of bacteria...... in dental biofilms. As their bactericidal action impacts the entire oral microflora, agents that inhibit biofilm formation without killing bacteria, such as the bovine milk protein osteopontin, have gained increasing attention. Here, we investigate the adhesion of 8 bacterial species associated with dental...... caries to salivary-coated flow-cells in the presence or absence of osteopontin or the control protein caseinoglycomacropeptide (0.32 mM/L). After 1h of flow (9.45 mm/min) at 35 °C, adhering bacteria were quantified by digital image analysis in a total of 692 bright-field images. Experiments were...

  7. STAT6-Dependent Collagen Synthesis in Human Fibroblasts Is Induced by Bovine Milk.

    Directory of Open Access Journals (Sweden)

    Stefan Kippenberger

    Full Text Available Since the domestication of the urus, 10.000 years ago, mankind utilizes bovine milk for different purposes. Besides usage as a nutrient also the external application of milk on skin has a long tradition going back to at least the ancient Aegypt with Cleopatra VII as a great exponent. In order to test whether milk has impact on skin physiology, cultures of human skin fibroblasts were exposed to commercial bovine milk. Our data show significant induction of proliferation by milk (max. 2,3-fold, EC50: 2,5% milk without toxic effects. Surprisingly, bovine milk was identified as strong inducer of collagen 1A1 synthesis at both, the protein (4-fold, EC50: 0,09% milk and promoter level. Regarding the underlying molecular pathways, we show functional activation of STAT6 in a p44/42 and p38-dependent manner. More upstream, we identified IGF-1 and insulin as key factors responsible for milk-induced collagen synthesis. These findings show that bovine milk contains bioactive molecules that act on human skin cells. Therefore, it is tempting to test the herein introduced concept in treatment of atrophic skin conditions induced e.g. by UV light or corticosteroids.

  8. STAT6-Dependent Collagen Synthesis in Human Fibroblasts Is Induced by Bovine Milk.

    Science.gov (United States)

    Kippenberger, Stefan; Zöller, Nadja; Kleemann, Johannes; Müller, Jutta; Kaufmann, Roland; Hofmann, Matthias; Bernd, August; Meissner, Markus; Valesky, Eva

    2015-01-01

    Since the domestication of the urus, 10.000 years ago, mankind utilizes bovine milk for different purposes. Besides usage as a nutrient also the external application of milk on skin has a long tradition going back to at least the ancient Aegypt with Cleopatra VII as a great exponent. In order to test whether milk has impact on skin physiology, cultures of human skin fibroblasts were exposed to commercial bovine milk. Our data show significant induction of proliferation by milk (max. 2,3-fold, EC50: 2,5% milk) without toxic effects. Surprisingly, bovine milk was identified as strong inducer of collagen 1A1 synthesis at both, the protein (4-fold, EC50: 0,09% milk) and promoter level. Regarding the underlying molecular pathways, we show functional activation of STAT6 in a p44/42 and p38-dependent manner. More upstream, we identified IGF-1 and insulin as key factors responsible for milk-induced collagen synthesis. These findings show that bovine milk contains bioactive molecules that act on human skin cells. Therefore, it is tempting to test the herein introduced concept in treatment of atrophic skin conditions induced e.g. by UV light or corticosteroids.

  9. Effects of bovine milk lactoperoxidase system on some bacteria.

    Science.gov (United States)

    Cankaya, M; Sişecioğlu, M; Bariş, O; Güllüce, M; Ozdemir, H

    2010-01-01

    Bovine lactoperoxidase (LPO) was purified from skimmed milk using amberlite CG-50-H+ resin, CM sephadex C-50 ion-exchange chromatography, and sephadex G-100 gel filtration chromatography. Lactoperoxidase was purified 20.45-fold with a yield of 28.8%. Purity of enzyme checked by sodium dodecyl sulphate-polyacrylamide gel electrophoresis method and a single band was observed. Km was 0.25 mM at 20 degrees C, Vmax value was 7.95 micromol/ml min at 20 degrees C (pH 6.0). Antibacterial study was done by disk diffusion method of Kir-by-Bauer using Mueller-Hinton agar medium with slight modification. Bovine LPO showed high antibacterial activity in 100 mM thiocyanate-100 mM H2O2 medium for some bacteria (Brevibacillus centrosaurus, B. choshinensis, B. lyticum, Cedecea davisae, Chryseobacterium indoltheticum, Clavibacter michiganense pv. insidiosum, Kocuria erythromyxa, K. kristinae, K. rosea, K. varians, Paenibacillus validus, Pseudomonas syringae pv. populans, Ralstonia pickettii, Rhodococcus wratislaviensis, Serratia fonticola, Streptomyces violaceusniger, Vibrio cholerae-nonO1) respectively, and compared with well known antibacterial substances (levofloxacin, netilmicin). LPO system has inhibition effects on all type bacteria and concentration is really important such as LPO-100 mM thiocyanate-100 mM H2O2 system was proposed as an effective agent against many factors causing several diseases.

  10. Effect of Lenient Steam Injection (LSI) heat treatment of bovine milk on the activities of some enzymes, the milk fat globule and pH

    DEFF Research Database (Denmark)

    Dickow, Jonatan A.; Nielsen, Martin Thorup; Hammershøj, Marianne

    2012-01-01

    This study investigated the effects of Lenient Steam Injection (LSI) treatment at temperatures 70–150 C on the enzymatic activities of the indigenous milk enzymes alkaline phosphatase, lactoperoxidase (LPO), xanthine oxidase (XO), lipoprotein lipase (LPL) and plasmin in comparison with two...

  11. Pellet-free isolation of human and bovine milk extracellular vesicles by size-exclusion chromatography

    Science.gov (United States)

    Blans, Kristine; Hansen, Maria S.; Sørensen, Laila V.; Hvam, Michael L.; Howard, Kenneth A.; Möller, Arne; Wiking, Lars; Larsen, Lotte B.; Rasmussen, Jan T.

    2017-01-01

    ABSTRACT Studies have suggested that nanoscale extracellular vesicles (EV) in human and bovine milk carry immune modulatory properties which could provide beneficial health effects to infants. In order to assess the possible health effects of milk EV, it is essential to use isolates of high purity from other more abundant milk structures with well-documented bioactive properties. Furthermore, gentle isolation procedures are important for reducing the risk of generating vesicle artefacts, particularly when EV subpopulations are investigated. In this study, we present two isolation approaches accomplished in three steps based on size-exclusion chromatography (SEC) resulting in effective and reproducible EV isolation from raw milk. The approaches do not require any EV pelleting and can be applied to both human and bovine milk. We show that SEC effectively separates phospholipid membrane vesicles from the primary casein and whey protein components in two differently obtained casein reduced milk fractions, with one of the fractions obtained without the use of ultracentrifugation. Milk EV isolates were enriched in lactadherin, CD9, CD63 and CD81 compared to minimal levels of the EV-marker proteins in other relevant milk fractions such as milk fat globules. Nanoparticle tracking analysis and electron microscopy reveals the presence of heterogeneous sized vesicle structures in milk EV isolates. Lipid analysis by thin layer chromatography shows that EV isolates are devoid of triacylglycerides and presents a phospholipid profile differing from milk fat globules surrounded by epithelial cell plasma membrane. Moreover, the milk EV fractions are enriched in RNA with distinct and diverging profiles from milk fat globules. Collectively, our data supports that successful milk EV isolation can be accomplished in few steps without the use of ultracentrifugation, as the presented isolation approaches based on SEC effectively isolates EV in both human and bovine milk. PMID:28386391

  12. An interactomics overview of the human and bovine milk proteome over lactation

    NARCIS (Netherlands)

    Zhang, Lina; Dijk, van Aalt-Jan; Hettinga, Kasper

    2017-01-01

    Background: Milk is the most important food for growth and development of the neonate, because of its nutrient composition and presence of many bioactive proteins. Differences between human and bovine milk in low abundant proteins have not been extensively studied. To better understand the differ

  13. Proteomic study on the stability of proteins in bovine, camel, and caprine milk sera after processing

    NARCIS (Netherlands)

    Zhang, Lina; Boeren, Sjef; Smits, Marcel; Hooijdonk, van Toon; Vervoort, Jacques; Hettinga, Kasper

    2016-01-01

    Milk proteins have been shown to be very sensitive to processing. This study aims to investigate the changes of the bovine, camel, and caprine milk proteins after freezing, pasteurization (62 °C, 30 min), and spray drying by proteomic techniques, filter-aided sample preparation (FASP) and dimethy

  14. Effect of soy and bovine milks on the dental plaque pH

    Directory of Open Access Journals (Sweden)

    Peyvand Moeiny

    2016-11-01

    (P<0.05. Conclusion: Bovine milk in none of frequent timing dropped its pH below basic pH but also significantly increased the plaque pH above the critical pH. The maximum pH drop for soy milk was in 2minutes after consumption but it never reached below the critical pH.

  15. Cell Infectivity in relation to bovine leukemia virus gp51 and p24 in bovine milk exosomes.

    Directory of Open Access Journals (Sweden)

    Tetsuya Yamada

    Full Text Available Exosomes are small membranous microvesicles (40-100 nm in diameter and are extracellularly released from a wide variety of cells. Exosomes contain microRNA, mRNA, and cellular proteins, which are delivered into recipient cells via these exosomes, and play a role in intercellular communication. In bovine leukemia virus (BLV infection of cattle, although it is thought to be a minor route of infection, BLV can be transmitted to calves via milk. Here, we investigated the association between exosomes and BLV in bovine milk. BLV structural proteins, gp51 (Env and p24 (Gag, were detected in bovine milk exosomes from BLV-infected cattle by Western blot analysis. In cells inoculated with these milk exosomes, BLV DNA was not detected during three serial passages by nested PCR. Purification of exosomes from persistently BLV-infected cells was achieved by immuno-magnetic separation using an antibody against exosomes coupled to magnetic beads. Consistently, BLV gp51 and p24 proteins were detected in purified exosomes. Moreover, reverse transcriptase activity was observed in purified exosomes, meaning that exosomes also contain viral enzyme. However, BLV DNA was not detected in serially passaged cells after inoculation of purified exosomes, indicating that exosomes carrying BLV proteins appeared to be not infectious. These results suggest that BLV proteins are released with milk exosomes and could be transferred into recipient cells of calves via milk exosomes as an alternative route not requiring virus infection. Moreover it is also possible that bovine milk exosomes play a role in clearance of BLV proteins from infected cells.

  16. Raw bovine milk improves gut responses to feeding relative to infant formula in preterm piglets.

    Science.gov (United States)

    Li, Yanqi; Jensen, Mikkel L; Chatterton, Dereck E W; Jensen, Bent B; Thymann, Thomas; Kvistgaard, Anne S; Sangild, Per T

    2014-01-01

    For preterm neonates, the quality of the first milk is crucial for intestinal maturation and resistance to necrotizing enterocolitis (NEC). Among other factors, milk quality is determined by the stage of lactation and processing. We hypothesized that unprocessed mature bovine milk (BM; raw bovine milk) would have less bioactivity than corresponding bovine colostrum (BC) in a preterm pig model, but have improved bioactivity relative to its homogenized, pasteurized, spray-dried equivalent, whole milk powder (WMP), or a bovine milk protein-based infant formula (IF). For 5 days, newborn preterm pigs received parenteral and enteral nutrition consisting of IF (n = 13), BM (n = 13), or BC (n = 14). In a second study, WMP (n = 15) was compared with IF (n = 10) and BM (n = 9). Compared with pigs fed IF, pigs that were fed BM had significantly improved intestinal structure (mucosal weight, villus height) and function (increased nutrient absorption and enzyme activities, decreased gut permeability, nutrient fermentation, and NEC severity). BC further improved these effects relative to BM (lactase activity, lactose absorption, plasma citrulline, and tissue interleukin-8). WMP induced similar effects as BM, except for lactase activity and lactose absorption. In conclusion, the maturational and protective effects on the immature intestine decreased in the order BC>BM>WMP, but all three intact bovine milk diets were markedly better than IF. The stage of lactation (colostrum vs. mature milk) and milk processing (e.g., homogenization, fractionation, pasteurization, spray-drying) are important factors in determining milk quality during the early postnatal period of preterm neonates.

  17. Sialyloligosaccharides in human and bovine milk and in infant formulas: variations with the progression of lactation.

    Science.gov (United States)

    Martín-Sosa, S; Martín, M J; García-Pardo, L A; Hueso, P

    2003-01-01

    Several lines of research support a role for human milk oligosaccharides in the defense of breast-fed infants against pathogens. Some ofthese oligosaccharides contain at least one moiety of sialic acid and are, thus, termed sialyloligosaccharides. These constitute a significant component (>1 g/L) of human milk. It is well established that milk composition varies among species, and previous reports have indicated that one ofthe differences between human and bovine milk is precisely their contents of sialyloligosaccharides. Because most infant formulas are manufactured with bovine milk components, it follows that formula-fed and breast-fed infants ingest dissimilar quantities of these carbohydrate structures. To ascertain these differences and their impact along lactation, the contents of oligosaccharide-bound sialic acids and major sialyloligosaccharides in samples of human and bovine milk (obtained at different lactation stages) were determined. In addition, infant formulas were assayed for their sialyloligosaccharide contents. Seven sialyloligosaccharides were identified in human milk; namely, 3'-sialyl-3-fucosyllactose and sialyllacto-N-tetraoses (a and b+c), the predominant structures at all lactation stages. Five sialyloligosaccharides were identified in bovine milk, of which 6'-sialyllactosamine and 3'-sialyllactose were the most abundant. In addition, sialyloligosaccharides in human and bovine milk decreased along lactation, and infant formulas did not contain significant amounts of sialyloligosaccharides. The results point to the general conclusion that regarding both qualitative and quantitative aspects, milk from humans and cows and infant formulas have different oligosaccharide contents. In this sense, bottle-fed infants are subject to reduced sialyloligosaccharide intake as compared to breast-fed infants.

  18. Bovine milk as a source of functional oligosaccharides for improving human health.

    Science.gov (United States)

    Zivkovic, Angela M; Barile, Daniela

    2011-05-01

    Human milk oligosaccharides are complex sugars that function as selective growth substrates for specific beneficial bacteria in the gastrointestinal system. Bovine milk is a potentially excellent source of commercially viable analogs of these unique molecules. However, bovine milk has a much lower concentration of these oligosaccharides than human milk, and the majority of the molecules are simpler in structure than those found in human milk. Specific structural characteristics of milk-derived oligosaccharides are crucial to their ability to selectively enrich beneficial bacteria while inhibiting or being less than ideal substrates for undesirable and pathogenic bacteria. Thus, if bovine milk products are to provide human milk-like benefits, it is important to identify specific dairy streams that can be processed commercially and cost-effectively and that can yield specific oligosaccharide compositions that will be beneficial as new food ingredients or supplements to improve human health. Whey streams have the potential to be commercially viable sources of complex oligosaccharides that have the structural resemblance and diversity of the bioactive oligosaccharides in human milk. With further refinements to dairy stream processing techniques and functional testing to identify streams that are particularly suitable for enriching beneficial intestinal bacteria, the future of oligosaccharides isolated from dairy streams as a food category with substantiated health claims is promising.

  19. Confirmatory analysis of firocoxib in bovine milk by rapid resolution liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Dowling, Geraldine; Gallo, Pasquale; Regan, Liam

    2009-02-15

    A rapid method has been developed to analyse for firocoxib (FIRO) residue in bovine milk. Milk samples were extracted with acetonitrile and sample extracts were purified on Evolute ABN solid phase extraction cartridges. Aliquots were analysed by rapid resolution liquid chromatography tandem mass spectrometry (RRLC-MS/MS). The method was validated in bovine milk, according to the criteria defined in Commission Decision 2002/657/EC. The decision limit (CCalpha) was 1.18ng/mL and for the detection capability a (CCbeta) value of 2.02ng/mL was obtained. The measurement uncertainty of the method was 27%. Fortifying bovine milk samples (n=18) in three separate assays, show the accuracy of the method to be between 96 and 105%. The precision of the method, expressed as RSD values for the within-lab reproducibility at the three levels of fortification (5, 7.5 and 10ng/mL) was less than 11% respectively.

  20. Doxorubicin-Induced Gut Toxicity in Piglets fed Bovine Milk and Colostrum

    DEFF Research Database (Denmark)

    Shen, René Liang; Rathe, Mathias; Jiang, Pingping

    2016-01-01

    a single dose of doxorubicin (1 × 75 mg/m) in piglets fed bovine milk enriched with whey protein (BM). In Study 2, responses to doxorubicin treatment were investigated in piglets receiving either seven BC feedings per day (Only-BC, n = 13), four BC feedings (High-BC, n = 13), two BC feedings (Low-BC, n...... milk-fed piglets suggesting that colostrum and a bovine milk diet enriched with whey protein provided similar......OBJECTIVE: Chemotherapy-induced intestinal toxicity is a common adverse effect of cancer treatment. We hypothesized that a milk diet containing bovine colostrum (BC) would reduce intestinal toxicity in doxorubicin-treated piglets. METHODS: Study 1 investigated intestinal parameters nine days after...

  1. Transcriptome profiling of bovine milk oligosaccharide metabolism genes using RNA-sequencing.

    Directory of Open Access Journals (Sweden)

    Saumya Wickramasinghe

    Full Text Available This study examines the genes coding for enzymes involved in bovine milk oligosaccharide metabolism by comparing the oligosaccharide profiles with the expressions of glycosylation-related genes. Fresh milk samples (n = 32 were collected from four Holstein and Jersey cows at days 1, 15, 90 and 250 of lactation and free milk oligosaccharide profiles were analyzed. RNA was extracted from milk somatic cells at days 15 and 250 of lactation (n = 12 and gene expression analysis was conducted by RNA-Sequencing. A list was created of 121 glycosylation-related genes involved in oligosaccharide metabolism pathways in bovine by analyzing the oligosaccharide profiles and performing an extensive literature search. No significant differences were observed in either oligosaccharide profiles or expressions of glycosylation-related genes between Holstein and Jersey cows. The highest concentrations of free oligosaccharides were observed in the colostrum samples and a sharp decrease was observed in the concentration of free oligosaccharides on day 15, followed by progressive decrease on days 90 and 250. Ninety-two glycosylation-related genes were expressed in milk somatic cells. Most of these genes exhibited higher expression in day 250 samples indicating increases in net glycosylation-related metabolism in spite of decreases in free milk oligosaccharides in late lactation milk. Even though fucosylated free oligosaccharides were not identified, gene expression indicated the likely presence of fucosylated oligosaccharides in bovine milk. Fucosidase genes were expressed in milk and a possible explanation for not detecting fucosylated free oligosaccharides is the degradation of large fucosylated free oligosaccharides by the fucosidases. Detailed characterization of enzymes encoded by the 92 glycosylation-related genes identified in this study will provide the basic knowledge for metabolic network analysis of oligosaccharides in mammalian milk. These candidate

  2. Effects of Diets Rich in Flavonoids on the Quality of Bovine Milk

    Directory of Open Access Journals (Sweden)

    Anastasia Cruz Carrillo

    2015-12-01

    Full Text Available Bovine milk is one of the main foods for mammals and, specifically, for humans; therefore, its quality should be optimal and should comply with all safety and nutritional aspects. The objective of this review was to collect updated scientific information on the importance of the presence of flavonoids in bovine milk, regarding their nutritional value and the benefits of these metabolites for the milk and its consumers. Milk decomposition is due to the effect of light, temperature and time on the fat and protein present in it, as well as to lactic fermentation by bacteria. Thus, the control of these three factors or one of them contributes to extend the useful life of milk and other dairy products. Thus, by seeking to delay the rancidification of milk and its by-products, it has been proposed that forage containing antioxidants such as flavonoids should be included in the diet of female cattle, as these, being bioavailable in cattle, also actively appear in the milk in concentrations equivalent to plasma. Additionally, the presence of flavonoids in bovine milk can provide a source of natural antioxidants for the consumer, which is beneficial for the prevention of some pathological processes mediated by free radicals.

  3. Mid-infrared predictions of cheese yield from bovine milk

    OpenAIRE

    Vanlierde, Amélie; Soyeurt, Hélène; Anceau, Christine; Vanden Bossche, sandrine; Dehareng, Frédéric; Pierre DARDENNE; Gengler, Nicolas; Sindic, Marianne; Colinet, Frédéric

    2011-01-01

    Economically, cheese yield (CY) is very important. Todate, empirical or theoretical formulae allow estimating the theoretical CY from milk fat and casein or protein content of milk. It would be interesting to predict CY during milk recording directly without the need to estimate milk components. Through the BlueSel project, 157 milk samples were collected in Wallonia from individual cows and analyzed using a mid-infrared (MIR) MilkoScanFT6000 spectrometer. Individual laboratory cheese yields ...

  4. Milk losses due to bovine tropical theileriosis (Theileria annulata infection) in Algeria

    Institute of Scientific and Technical Information of China (English)

    Ouarda Ayadi; Mohamed Gharbi

    2016-01-01

    The authors studied the impact of tropical theileriosis onset on milk yield decrease in 10 local bred cows in Skikda (Northern Algeria) during 2015 summer season. The milk yield decrease estimated weekly during two months was 2.76 L/day/cow corresponding to 31.92%of the total milk yield. This decrease corresponds to 110.5 Algerian Dinars (1.02 US$)/day/diseased cow. The relative variation of milk yield showed a dramatic decrease from 82.72% to 0.76% at Day 21 then became constant. Further studies are needed to improve these estimations of financial losses due to bovine tropical theileriosis in Algeria.

  5. Pheno- and genotyping of Staphylococcus epidermidis isolated from bovine milk and human skin

    DEFF Research Database (Denmark)

    Thorberg, B. M.; Kuhn, I.; Aarestrup, Frank Møller

    2006-01-01

    The purpose of this study was to improve our knowledge concerning the epidemiology and strain diversity of Staphylococcus epidermidis isolated from bovine milk in commercial dairy herds. A total of 341 S. epidermidis isolates obtained from cows' milk (317), farmers (17) and patients (7) were...... different S. epidermidis types exist in milk samples. Antibiotic resistance patterns matched the SmaI profiles closely in the two herds, but poorly in the routinely collected milk samples. Isolates from herd I showed one to five patterns, depending on the typing method used. Isolates from the milker's skin...

  6. External cavity-quantum cascade laser (EC-QCL) spectroscopy for protein analysis in bovine milk.

    Science.gov (United States)

    Kuligowski, Julia; Schwaighofer, Andreas; Alcaráz, Mirta Raquel; Quintás, Guillermo; Mayer, Helmut; Vento, Máximo; Lendl, Bernhard

    2017-04-22

    The analytical determination of bovine milk proteins is important in food and non-food industrial applications and yet, rather labour-intensive wet-chemical, low-throughput methods have been employed since decades. This work proposes the use of external cavity-quantum cascade laser (EC-QCL) spectroscopy for the simultaneous quantification of the most abundant bovine milk proteins and the total protein content based on the chemical information contained in mid-infrared (IR) spectral features of the amide I band. Mid-IR spectra of protein standard mixtures were used for building partial least squares (PLS) regression models. Protein concentrations in commercial bovine milk samples were calculated after chemometric compensation of the matrix contribution employing science-based calibration (SBC) without sample pre-processing. The use of EC-QCL spectroscopy together with advanced multivariate data analysis allowed the determination of casein, α-lactalbumin, β-lactoglobulin and total protein content within several minutes.

  7. Enzymatic-fluorometric quantification of cholesterol in bovine milk

    DEFF Research Database (Denmark)

    Larsen, Torben

    2012-01-01

    The present paper describes an enzymatic–fluorometric method for the determination of cholesterol in milk and other opaque matrices. The initial step of the method is to liberate chemically and physically bound cholesterol from the milk fat globule membrane by enzymatic action. The method is able...... to discriminate between esterified and free cholesterol in milk. The analysis is cost effective and is developed to work directly on whole, fresh milk thereby eliminating time consuming and tedious pre-treatment procedures of the sample. More than 1000 milk samples were analysed on the day of sampling. The total...... concentration of milk cholesterol ranged from 80 to 756 μM (n = 1068; mean 351 μM). Milk cholesterol was significantly correlated to milk fat concentration as analysed by mid-infra red spectrometry (r = 0.630; n = 853) and by an enzymatic–fluorometric method (triacylglycerol) (r = 0.611; n = 842)....

  8. The Host Defense Proteome of Human and Bovine Milk

    NARCIS (Netherlands)

    Hettinga, K.A.; Valenberg, van H.J.F.; Vries, de S.C.; Boeren, S.; Hooijdonk, van A.C.M.; Arendonk, van J.A.M.; Vervoort, J.J.M.

    2011-01-01

    Milk is the single source of nutrients for the newborn mammal. The composition of milk of different mammals has been adapted during evolution of the species to fulfill the needs of the offspring. Milk not only provides nutrients, but it also serves as a medium for transfer of host defense components

  9. Comparison of the activity of human and bovine milk on two cell lines.

    Science.gov (United States)

    Pocoví, Coloma; Conesa, Celia; Barbana, Chockry; Pérez, María D; Calvo, Miguel; Sánchez, Lourdes

    2009-08-01

    The activity of human milk on cell growth has been evaluated on two cell lines, MDCK and Caco-2. The proportion of human milk samples that reduced by half the growth of MDCK cells was of 36%. This inhibitory activity was associated with casein and not the whey fraction. Great variability was found in the degree of inhibitory activity depending on the milk sample. The susceptibility of Caco-2 cells to milk inhibitory activity was lower than that of MDCK. Bovine milk did not have any effect on cell growth, either as skimmed milk or as whey or casein. Morphology of cells incubated with active human casein showed abnormal features, such as chromatin condensation, reduced cellular volume and apoptotic bodies, and also fragmented DNA, which are all features of apoptosis.

  10. Effect on milk production of vaccination with a bovine herpesvirus 1 gene-deleted vaccine.

    Science.gov (United States)

    Bosch, J C; Frankena, K; van Oirschot, J T

    1997-02-22

    A field trial was conducted to determine the effect of vaccination with an inactivated bovine herpesvirus 1 (BHV1) gE-negative marker vaccine on the milk production of dairy cows. The daily milk yield of 455 cows in six herds was measured electronically from six days before vaccination until 14 days after vaccination. The treatment consisted of two injections with either vaccine or placebo, both at an interval of four weeks. There was a small, but significant (P < 0.05), decrease of about 1.4 litres per cow in milk production after a double vaccination, the negative effect being slightly greater after the second vaccination.

  11. Exploration of bovine milk proteome in colostral and mature whey using an ion-exchange approach.

    Science.gov (United States)

    Le, Anh; Barton, L Douglas; Sanders, Jeff T; Zhang, Qiang

    2011-02-04

    In addition to milk's nutritional role, it contains immunoglobulins (antibodies) and immunoregulatory proteins that are active in the digestive tract of newborns. However, knowledge of the repertoire of milk proteins remains meager. In this work, we report an ion-exchange-based protein fractionation method that allows in-depth exploration of the whey proteome in bovine milk; 293 unique gene products were identified, of which 176 were newly identified in whey. This work also demonstrated qualitatively for the first time the consistency, albeit differing in protein levels, in milk proteome between colostrum and mature milk (3 mo. post calving). Semiquantitative analysis showed a number of up-regulated proteins in colostrum that may provide extra natural defenses for the neonate. Increased understanding of the composition and functions of bovine milk proteins and their potential health benefits may, in the future, play an important role in nutritional and biomedical applications as properly processed cow's milk proteins could potentially confer the same bioactivity as their human counterparts.

  12. Stochastic simulation modeling to determine time to detect Bovine Viral Diarrhea antibodies in bulk tank milk

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Enøe, Claes; Krogh, Kaspar;

    2014-01-01

    A stochastic simulation model was developed to estimate the time from introduction ofBovine Viral Diarrhea Virus (BVDV) in a herd to detection of antibodies in bulk tank milk(BTM) samples using three ELISAs. We assumed that antibodies could be detected, after afixed threshold prevalence...

  13. Gold nanoparticle aggregation-based colorimetric assay for β-casein detection in bovine milk samples.

    Science.gov (United States)

    Li, Y S; Zhou, Y; Meng, X Y; Zhang, Y Y; Song, F; Lu, S Y; Ren, H L; Hu, P; Liu, Z S; Zhang, J H

    2014-11-01

    Traditional Kjeldahl method, used for quality evaluation of bovine milk, has intrinsic defects of time-consuming sample preparation and two analyses to determine the difference between non-protein nitrogen content and total protein nitrogen content. Herein, based upon antibody functionalized gold nanoparticles (AuNPs), we described a colorimetric method for β-casein (β-CN) detection in bovine milk samples. The linear dynamic range and the LOD were 0.08-250 μg mL(-1), and 0.03 μg mL(-1) respectively. In addition, the real content of β-CN in bovine milk was measured by using the developed assay. The results are closely correlated with those from Kjeldahl method. The advantages of β-CN triggered AuNP aggregation-based colorimetric assay are simple signal generation, the high sensitivity and specificity as well as no need of complicated sample preparation, which make it for on-site detection of β-CN in bovine milk samples.

  14. Rapid determination of ampicillin in bovine milk by liquid chromatography with fluorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Ang, C.Y.W.; Luo, Wenhong [National Center for Toxicological Research, Jefferson, AR (United States)

    1997-01-01

    A rapid and sensitive liquid chromatographic (LC) method was developed for the determination of ampicillin residues in raw bovine milk, processed skim milk, and pasteurized, homogenized whole milk with vitamin D. Milk samples were deproteinized with trichloroacetic acid (TCA) and acetonictrile. After centrifugation, the clear supernatant was reacted with formaldehyde and TCA under heat. The major fluorescent derivative of ampicillin was then determined by reversed-phase LC with fluorescence detection. Average recoveries of ampicillin fortified at 5, 10, and 20 ppb (ng/mL) were all >85% with coefficients of variation <10%. Limits of detection ranged from 0.31 to 0.51 ppb and limits of quantitation, from 0.66 to 1.2 ppb. After appropriate validation, this method should be suitable for rapid analysis of milk for ampicillin residues at the tolerance level of 10 ppb. 16 refs., 4 figs., 3 tabs.

  15. Acute phase proteins in bovine milk in an experimental model of Staphylococcus aureus subclinical mastitis

    DEFF Research Database (Denmark)

    Eckersall, P D; Young, F J; Nolan, A M

    2006-01-01

    The objectives were to establish the origin of 2 acute phase proteins in milk during subclinical bovine mastitis and to characterize the relationship between those proteins in milk and blood. Haptoglobin (Hp) and mammary-associated serum amyloid A (M-SAA3) appear in milk during mastitis, whereas Hp...... and serum amyloid A increase in serum during mastitis. The concentrations of these proteins were determined in an experimental model using a field strain of Staphylococcus aureus to induce subclinical mastitis in dairy cows. The expression of mRNA coding for these proteins was assessed and the presence of M......-SAA3 in mammary tissues was determined using immunocytochemistry. Increases of M-SAA3 and Hp in milk occurred within 12 h of Staphylococcus aureus infusion, with peak concentrations occurring 3 d after infusion of the bacteria. The increase of acute phase proteins in milk (15 h) preceded the increase...

  16. Genetic and environmental variation in bovine milk infrared spectra

    NARCIS (Netherlands)

    Wang, Qiuyu; Hulzebosch, Alex; Bovenhuis, Henk

    2016-01-01

    Fourier transform infrared (FTIR) spectroscopy is widely used to determine milk composition. In this study, 1,060 milk infrared wavenumbers ranging from 925 to 5,008 cm-1 of 1,748 Holstein Friesian cows on 371 herds in the Netherlands were available. The extent to which infrared wavenu

  17. Effect of continuous milking on immunoglobulin concentrations in bovine colostrum

    NARCIS (Netherlands)

    Verweij, J.J.; Koets, A.P.; Eisenberg, S.W.F.

    2014-01-01

    Continuous milking is defined as a dairy cattle management system without a planned dry period for cows in late gestation. Continuous milking has been described to reduce health problems common in periparturient cattle, but may affect colostrum immunoglobulin (Ig) concentration and subsequently calf

  18. Toward a dry reagent immunoassay of progesterone in bovine milk

    NARCIS (Netherlands)

    Posthuma-Trumpie, Geertruida Afina

    2008-01-01

    This thesis is aimed at the development of a dry reagent immunoassay of progesterone in cow's milk. Progesterone is a steroid hormone and regulates ovulation in female mammals. The concentration of progesterone in blood and in milk is in accordance with the reproductive cycle of the individual femal

  19. Adhesive properties of predominant bacteria in raw cow's milk to bovine mammary gland epithelial cells.

    Science.gov (United States)

    Hagi, Tatsuro; Sasaki, Keisuke; Aso, Hisashi; Nomura, Masaru

    2013-11-01

    Various bacteria have been found in raw cow's milk, and identifying milk microflora and its functions is critical for maintaining cow health and farm hygiene. Although studies on pathogens and spoilage bacteria in milk have been widely reported, the relationship between milk bacteria, including nonpathogenic bacteria, and the bovine udder is poorly understood. We investigated milk microflora over 1 year using a culture-dependent method and culture-independent analysis by denaturing gradient gel electrophoresis. Among 240 isolates, Lactococcus lactis (81/240) was predominant. The predominant genera were Lactococcus, Stenotrophomonas, Microbacterium, Chryseobacterium, Serratia and Pseudomonas. Among seven strains belonging to these predominant genera, two strains of L. lactis (ssp. lactis and ssp. cremoris) exhibited the highest adherence to bovine mammary gland epithelial cells (BMECs) derived from the bovine udder; 3.4 % of the inoculated bacteria adhered to BMECs. This was followed by Serratia sp. (1.6 %), Microbacterium sp. (0.8 %), Stenotrophomonas maltophilia (0.5 %), Pseudomonas sp. (0.3 %) and Chryseobacterium sp. (0.1 %). The two L. lactis isolates exhibited higher adherence to BMECs than type strains and isolates of various origins.

  20. Concentrations of sialyloligosaccharides in bovine colostrum and milk during the prepartum and early lactation.

    Science.gov (United States)

    Nakamura, T; Kawase, H; Kimura, K; Watanabe, Y; Ohtani, M; Arai, I; Urashima, T

    2003-04-01

    Sialyloligosaccharides and sialylglycoconjugates in colostrum and milk are regarded to be important biological components with respect to be source of brain gangliosides in infant and to be antiinfectional components for the attack by the pathogenic bacteria and virus. Several acidic oligosaccharides have been characterised in both bovine and human milk or colostrum. The sialyloligosaccharide content of human colostrum and milk has been extensively studied, whereas that of cows milk and colostrum has received less attention. In this study, the concentrations of three sialyloligosaccharides of bovine colostrum and milk were determined at various stages during the prepartum and the first 7 d postpartum. The concentration of 3'SL (Neu5Ac(alpha2-3)Gal(beta1-4)Glc) reached a maximum value of 0.85 mg/ml immediately following parturition while the concentrations of 6'SL (Neu5Ac(alpha2-6)Gal(beta1-4)Glc) and 6'SLN (Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc) of 0.14 and 0.12 mg/ml, respectively, were much lower at this initial stage, although these concentration were maximum immediately following parturition. Bovine colostrum, especially that collected immediately after parturition, may be suitable as a source of 3'SL and other sialyloligosaccharides for use as additives by the food or pharmaceutical industries.

  1. A method for routine estimation of vitamin D activity in human and bovine milk.

    Science.gov (United States)

    Parviainen, M T; Koskinen, T; Ala-Houhala, M; Visakorpi, J K

    1984-01-01

    To estimate the antirachitic activity of human and bovine milk, we developed a modern biochemical method for determining vitamin D metabolites in milk. Vitamin D metabolites were assayed from milk whey and from whole milk. Milk whey yielded poor recovery of both endogenous and added vitamin D, suggesting a marked loss of vitamin D activity to milk fat during homogenization and separation of the milk whey. A method for assaying the vitamin D metabolites in whole milk involves 1) lipid extraction, 2) cold methanol and ether precipitation, 3) alkaline backwash to reduce the amount of interfering lipids, 4) an efficient reverse-phase preparative purification, 5) an additional silica purification for vitamin D, 6) an analytical high-performance liquid chromatography, and 7) separate sensitized protein-binding assays for vitamin D and 25-hydroxyvitamin D. The method for whole milk resulted in good recovery of added vitamin D, and levels of assayed metabolites and their calculated antirachitic activity agreed well with earlier reports, that is, about 10-50 IU of vitamin D activity per liter.

  2. Bovine respiratory syncytial virus reinfections and decreased milk yield in dairy cattle.

    Science.gov (United States)

    van der Poel, W H; Mourits, M C; Nielen, M; Frankena, K; Van Oirschot, J T; Schukken, Y H

    1995-09-01

    The influence of Bovine Respiratory Syncytial Virus (BRSV) reinfections on the daily milk yield was studied by evaluating the milk production of 32 BRSV reinfected cows. For the estimation of milk production losses, four lactation curve models were used, including a gamma function, a second degree polynomial, and both of these models with a lag variable. Bovine respiratory syncytial virus reinfections seemed to have only a small effect on the daily milk production. Comparison of the true production with an estimated production according to the gamma function showed that the production for first lactation cows dropped 0.14 kg on average and for cows in their second or later lactation 0.56 kg on average, during 5 consecutive days in the infection period. For the second-degree polynomial model these values were respectively 0.42 kg and 0.80 kg. All calculated average production losses were relatively small and not significant (P > 0.15). The models without lag variable were more suitable than the models with the lag variable to estimate small production losses caused by BRSV reinfections. The power of this study was sufficient to detect a decrease in production of approximately 1-1.5 kg milk per cow per day. It was therefore concluded that BRSV reinfections were not associated with an important loss of milk production.

  3. Limit of detection of Toxocara canis larvae in experimentally contaminated bovine milk

    Directory of Open Access Journals (Sweden)

    Edlayne Larissa Gretter Machado Pereira

    2016-02-01

    Full Text Available This study aimed to evaluate the limit of detection of Toxocara canis larvae in experimentally contaminated commercial bovine milk samples, based on a centrifuge-sedimentation technique. Firstly, bovine milk (whole and skim samples were contaminated with 50 T. canis larvae in order to evaluate the interference of milk fat with the recovery of the larvae. Next, the effects of 10% formalin (100 ?L, ether (100 ?L, and a combination of both solutions on the recovery of the larvae was examined. Thereafter, the limit of detection of the larvae was determined using the solution (from step 2 considered optimal for degreasing the milk sample. Samples were contaminated with aliquots of 1, 5, 10, 25, and 50 larvae. For each milk sample (1.0 mL, 15 repetitions were analysed. The recovery of the larvae from the skim milk samples was higher (p = 0.0031 than that from the whole milk samples. No significant difference (p = 0.5681 was observed with regard to the percentage of recovered larvae when comparing the degreasing solutions. Nevertheless, the formalin-ether combination was more efficient for recovering the larvae (73.1% than ether (71.9%, formalin (67.6%, and pure whole milk (70.0%. Concerning the limit of detection (using formalin-ether, all the samples contaminated with 5, 10, 25, and 50 larvae tested positive (minimum: 62.7%. Of the samples contaminated with a single larva, 66.7% tested positive. These results suggest that the centrifugation-sedimentation technique may be useful for recovering larvae of Toxocara spp. in naturally or experimentally contaminated milk samples obtained from a wide range of animal species.

  4. Determination of ivermectin and moxidecin residues in bovine milk and examination of the effects of these residues on acid fermentation of milk.

    Science.gov (United States)

    Imperiale, F; Sallovitz, J; Lifschitz, A; Lanusse, C

    2002-09-01

    Ivermectin (IVM) and moxidectin (MXD) are broad-spectrum antiparasitic drugs not approved for use in dairy animals, although their use in dairy sheep, goats and cattle nevertheless occurs in many parts of the world. The work reported here describes (1) the application of an HPLC method (including milk samples clean-up and chemical extraction) to quantify IVM and MXD residues in bovine milk, and (2) an assessment of the effect of different IVM and MXD concentrations on bovine milk acid fermentation. The latter was carried out using the 'yoghurt test' to determine the minimum IVM and MXD concentrations affecting milk acid fermentation. The sample clean-up, chemical extraction and the validated HPLC method allowed the quantification of IVM and MXD up to 0.1 ng ml(-1) in milk with acceptable validation coefficients. Drug recoveries from fortified milk samples ranged between 72% (CV = 9.1%) and 75% (CV = 13.3%) for MXD and IVM, respectively. Neither IVM nor MXD affected the acid fermentation of bovine milk. In fact, there was no drug-induced changes on milk acidity even at IVM and MXD concentrations as high as 1000 ng ml(-1). These results indicate that the yoghurt biological test is not suitable to evaluate the presence of milk residues for these antiparasitic compounds. Thus, a highly sensitive HPLC technique is the only reliable method for determining the presence of residual concentrations of IVM and MXD in milk and dairy products to assure consumer safety.

  5. 2d electrophoresis of bovine milk proteins and milk fermented drink

    OpenAIRE

    Damir Mogut; Anna Iwaniak; Monika Hrynkiewicz; Jerzy Dziuba

    2015-01-01

    The aim of the study was the analysis of milk and milk fermented drink proteomes with the use of 2D electrophoresis. The criteria of proteins separation were the values of their isoelectric points (pI) and molecular weights (MW). Our results showed that milk and milk fermented drink proteomes consisted of 118 and 121 spots, respectively. The computer analysis revealed the identity of 95 spots in both proteomes. Non-identical spots indicated the changes resulting from the action of bact...

  6. Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography.

    Science.gov (United States)

    Zhang, Yan; Luo, Jian; Bi, Jingxiu; Wang, Jun; Sun, Lijing; Liu, Yongdong; Zhang, Guifeng; Ma, Guanghui; Su, Zhiguo

    2010-06-04

    Transgenic bovine milk could be a rich source of recombinant human proteins. However, the co-presence of bovine and human homologous proteins can be a challenge for product purification. In this study, the average surface hydrophobicity and electric potential of human alpha-lactalbumin (HLA) and bovine alpha-lactalbumin (BLA) were analyzed and compared through the exposure area calculation of different amino acids. Based on the analysis, calcium independent hydrophobic interaction chromatography was selected for separation of recombinant human alpha-lactalbumin (rHLA) from BLA in transgenic bovine milk. The operating conditions for the best separation of two proteins were predicted by fluorescence data. Three commercially available HIC resins (Butyl Sepharose 4 FF, Octyl Sepharose 4 FF, Phenyl Sepharose 6 FF) were compared. The transgenic milk was skimmed and treated by pH adjustment to remove a large quantity of casein protein. The supernatant was loaded on the hydrophobic interaction chromatographic matrix. The correct elution fraction was further treated with gel filtration chromatography. The overall recovery of rHLA was up to 67.1% with the purity greater than 95%. Circular dichroism spectroscopy (CD) and mass spectrogram (MS) confirmed the native state and glycosylated form of the purified rHLA.

  7. Radioimmunoassay for progesterone in bovine milk; Radioinmunoensayo para progesterona en leche bovina

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz, Miriam [Instituto Superior de Ciencias y Tecnologia Nucleares, La Habana (Cuba)]. E-mail: mirian@fctn.isctn.edu.cu; Figueredo, Nancy; Castillo, Sonia; Pizarro [Centro de Isotopos, La Habana (Cuba)

    2002-07-01

    A system for the measurement of progesterone in bovine milk by radioimmunoassay has been developed and validated. This assay includes an iodine tracer purified by HPLC, the standard prepared in fat-free milk and an antibody anti-progesterone combined with second antibody. The detection limit of the assay is at 0.2 nmol/L calculated from the maximum binding menus two standard deviations and the precision is satisfactory. In the recovery assay was used 4 milk different samples and the result was 98% of recuperation. The progesterone was determinate in milk samples from post-partum animals taking samples three times per week for 40 days. The assay is simple, rapid and possibility the progesterone measurement without sample dilution, distinguish the cyclic changes of this hormone that reflect the ovarian activity in the animals. (author)

  8. Some factors influencing the syneresis of bovine, ovine, and caprine milks.

    Science.gov (United States)

    Calvo, M M; Balcones, E

    2000-08-01

    The influence of the species, fat, curd incubation temperature (25, 30, and 35 degrees C), heat treatment of milk (70 degrees C for 5 or 30 min), and milk pH on the initial volume drained, syneresis rate, and the loss of proteins in drainage was studied. The volume drained as a function of the curd incubation time (up to 60 min) was adjusted to a first-order kinetic reaction. The k values (drained rate) and the initial volume obtained applying the equation were compared to establish the possible influence of the studied factors. In general, for all the factors studied, the syneresis rate of curd from caprine and ovine milk did not differ from those described previously in the literature for the curds from bovine milk. However, for each studied factor the pattern of syneresis rate was significantly different among the species in most of the experiments.

  9. Milk losses due to bovine tropical theileriosis (Theileria annulata infection in Algeria

    Directory of Open Access Journals (Sweden)

    Ouarda Ayadi

    2016-09-01

    Full Text Available The authors studied the impact of tropical theileriosis onset on milk yield decrease in 10 local bred cows in Skikda (Northern Algeria during 2015 summer season. The milk yield decrease estimated weekly during two months was 2.76 L/day/cow corresponding to 31.92% of the total milk yield. This decrease corresponds to 110.5 Algerian Dinars (1.02 US$/day/diseased cow. The relative variation of milk yield showed a dramatic decrease from 82.72% to 0.76% at Day 21 then became constant. Further studies are needed to improve these estimations of financial losses due to bovine tropical theileriosis in Algeria.

  10. The influence of bovine milk high or low in isoflavones on hepatic gene expression in mice

    DEFF Research Database (Denmark)

    Skaanild, Mette Tingleff; Nielsen, Tina Skau

    2012-01-01

    in hepatic gene expression after dietary intake of milk high and low in isoflavones. In addition to pelleted feed female NMRI mice were offered water, water added either 17β-estradiol, equol, Tween 80, and milk high and low in isoflavone content for a week. Gene expression was analyzed using an array q......Isoflavones have generated much attention due to their potential positive effects in various diseases. Phytoestrogens especially equol can be found in bovine milk, as feed ration for dairy cows is comprised of plants containing phytoestrogens. The aim of this study was to analyze the changes......PCR kit. It was revealed that Tween 80 and 17β-estradiol upregulated both phase I and phase II genes to the same extent whereas equol alone, high and low isoflavone milk did not alter the expression of phase I genes but decreased the expression of phase II genes. This study shows that dietary isoflavones...

  11. Detection of Helicobacter pylori in raw bovine milk by fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Angelidis, Apostolos S; Tirodimos, Ilias; Bobos, Mattheos; Kalamaki, Mary S; Papageorgiou, Demetrios K; Arvanitidou, Malamatenia

    2011-12-02

    The transmission pathways of Helicobacter pylori in humans have not been fully elucidated. Research in the last decade has proposed that foodborne transmission, among others, may be a plausible route of human infection. Owing to the organism's fastidious growth characteristics and its ability to convert to viable, yet unculturable states upon exposure to stress conditions, the detection of H. pylori in foods via culture-dependent methods has been proven to be laborious, difficult and in most cases unsuccessful. Hence, nucleic acid-based methods have been proposed as alternative methods but, to date, only PCR-based methods have been reported in the literature. In the current study, fluorescence in situ hybridization (FISH) was used for the detection of H. pylori in raw, bulk-tank bovine milk. After repeated milk centrifugation and washing steps, the bacterial flora of raw milk was subjected to fixation and permeabilization and H. pylori detection was conducted by FISH after hybridization with an H. pylori-specific 16S rRNA-directed fluorescent oligonucleotide probe. Using this protocol, H. pylori was detected in four out of the twenty (20%) raw milk samples examined. The data presented in this manuscript indicate that FISH can serve as an alternative molecular method for screening raw bovine milk for the presence of H. pylori.

  12. Raw bovine milk improves gut responses to feeding relative to infant formula in preterm piglets

    DEFF Research Database (Denmark)

    Li, Yang; Lykke, Mikkel; Chatterton, D E W

    2014-01-01

    height) and function (increased nutrient absorption and enzyme activities, decreased gut permeability, nutrient fermentation, and NEC severity). BC further improved these effects relative to BM (lactase activity, lactose absorption, plasma citrulline, and tissue interleukin-8). WMP induced similar...... effects as BM, except for lactase activity and lactose absorption. In conclusion, the maturational and protective effects on the immature intestine decreased in the order BC>BM>WMP, but all three intact bovine milk diets were markedly better than IF. The stage of lactation (colostrum vs. mature milk...

  13. Effects of mineral content of bovine drinking water: does iron content affect milk quality?

    Science.gov (United States)

    Mann, G R; Duncan, S E; Knowlton, K F; Dietrich, A D; O'Keefe, S F

    2013-01-01

    The composition of water given to dairy cattle is often ignored, yet water is a very important nutrient and plays a major role in milk synthesis. The objective of this study was to study effects of elevated levels of iron in bovine drinking water on milk quality. Ferrous lactate treatments corresponding to 0, 2, 5, and 12.5mg/kg drinking water concentrations were delivered through the abomasum at 10 L/d to 4 lactating dairy cows over 4 periods (1 wk infusion/period) in a Latin square design. On d 6 of infusion, milk was collected, processed (homogenized, pasteurized), and analyzed. Mineral content (Fe, Cu, P, Ca) was measured by inductively coupled plasma mass spectrometry. Oxidative stability of whole processed milk was measured by the thiobarbituric acid reactive substances (TBARS) assay for malondialdehyde (MDA) and sensory analysis (triangle test) within 72 h of processing and after 7d of storage (4°C). Significant sensory differences between processed milks from cows receiving iron and the control infusion were observed. No differences in TBARS (1.46±0.04 mg of MDA/kg) or mineral content (0.22±0.01 mg/kg Fe) were observed. A 2-way interaction (iron treatment by cow) for Ca, Cu, and Fe concentrations was seen. While iron added directly to milk causes changes in oxidation of milk, high levels of iron given to cattle have subtle effects that initially may not be obvious.

  14. 2d electrophoresis of bovine milk proteins and milk fermented drink

    Directory of Open Access Journals (Sweden)

    Damir Mogut

    2015-09-01

    Full Text Available The aim of the study was the analysis of milk and milk fermented drink proteomes with the use of 2D electrophoresis. The criteria of proteins separation were the values of their isoelectric points (pI and molecular weights (MW. Our results showed that milk and milk fermented drink proteomes consisted of 118 and 121 spots, respectively. The computer analysis revealed the identity of 95 spots in both proteomes. Non-identical spots indicated the changes resulting from the action of bacterial proteinases on milk proteins during the production of milk fermented drink. Proteolytic starter cultures applied to produce the milk fermented drink led to a partial hydrolysis of milk proteins to peptides and free amino acid residues. Results obtained led to confirm the suitability of 2D electrophoresis to observe the changes in the proteomes of milk products, as well as other food products after the application of technological processes. The development of the methods of proteomic analysis will let, in the future, eliminate the artefacts, which may limit the possibility of errors during proteins’ identification.

  15. Double-antibody based immunoassay for the detection of β-casein in bovine milk samples.

    Science.gov (United States)

    Zhou, Y; Song, F; Li, Y S; Liu, J Q; Lu, S Y; Ren, H L; Liu, Z S; Zhang, Y Y; Yang, L; Li, Z H; Zhang, J H; Wang, X R

    2013-11-01

    The concentration of casein (CN) is one of the most important parameters for measuring the quality of bovine milk. Traditional approach to CN concentration determination is Kjeldahl, which is an indirect method for determination of total nitrogen content. Here, we described a double-antibody based direct immunoassay for the detection of β-CN in bovine milk samples. Monoclonal antibody (McAb) was used as capture antibody and polyclonal antibody (PcAb) labelled with horseradish peroxidase (HRP) as detection antibody. With the direct immunoassay format, the linear range of the detection was 0.1-10.0 μg mL(-1). The detection limit was 0.04 μg mL(-1). In addition, the concentration of β-CN in real bovine milk samples has been detected by the developed immunoassay. There was a good correlation between the results obtained by the developed technique and Kjeldahl method from commercial samples. Compared to the traditional approach, the advantage of the assay is no need of time-consuming sample pretreatment.

  16. A prospective study of cow's milk allergy in exclusively breast-fed infants. Incidence, pathogenetic role of early inadvertent exposure to cow's milk formula, and characterization of bovine milk protein in human milk

    DEFF Research Database (Denmark)

    Høst, A; Husby, S; Osterballe, O

    1988-01-01

    with ELISA indicated a molecular weight of BLG corresponding to that of monomeric BLG (18 kD). Possibly early inadvertent and occasional exposure to cow's milk proteins may initiate sensitization in predisposed neonates. Subsequent exposure to minute amounts of bovine milk proteins in human milk may act......A cohort of 1,749 newborns in the municipality of Odense were followed prospectively for the development of cow's milk allergy (CMA) during their first year of life. Altogether 39 fulfilled the criteria for CMA (2.2%). Out of the 39 infants, 17 developed symptoms of CMA during breast...... had signs of CMA in the neonatal period. Review of records from the newborn nursery revealed that all 9 infants had been exposed to cow's milk formula in amounts corresponding to approximately 0.4-3.0 g of beta-lactoglobulin (BLG) during the first three days of life. Human milk samples were analyzed...

  17. Mid-infrared prediction of lactoferrin content in bovine milk: potential indicator of mastitis.

    Science.gov (United States)

    Soyeurt, H; Bastin, C; Colinet, F G; Arnould, V M-R; Berry, D P; Wall, E; Dehareng, F; Nguyen, H N; Dardenne, P; Schefers, J; Vandenplas, J; Weigel, K; Coffey, M; Théron, L; Detilleux, J; Reding, E; Gengler, N; McParland, S

    2012-11-01

    Lactoferrin (LTF) is a milk glycoprotein favorably associated with the immune system of dairy cows. Somatic cell count is often used as an indicator of mastitis in dairy cows, but knowledge on the milk LTF content could aid in mastitis detection. An inexpensive, rapid and robust method to predict milk LTF is required. The aim of this study was to develop an equation to quantify the LTF content in bovine milk using mid-infrared (MIR) spectrometry. LTF was quantified by enzyme-linked immunosorbent assay (ELISA), and all milk samples were analyzed by MIR. After discarding samples with a coefficient of variation between 2 ELISA measurements of more than 5% and the spectral outliers, the calibration set consisted of 2499 samples from Belgium (n = 110), Ireland (n = 1658) and Scotland (n = 731). Six statistical methods were evaluated to develop the LTF equation. The best method yielded a cross-validation coefficient of determination for LTF of 0.71 and a cross-validation standard error of 50.55 mg/l of milk. An external validation was undertaken using an additional dataset containing 274 Walloon samples. The validation coefficient of determination was 0.60. To assess the usefulness of the MIR predicted LTF, four logistic regressions using somatic cell score (SCS) and MIR LTF were developed to predict the presence of mastitis. The dataset used to build the logistic regressions consisted of 275 mastitis records and 13 507 MIR data collected in 18 Walloon herds. The LTF and the interaction SCS × LTF effects were significant (P test. In conclusion, this study confirms the possibility to quantify an LTF indicator from milk MIR spectra. It suggests the usefulness of this indicator associated to SCS to detect the presence of mastitis. Moreover, the knowledge of milk LTF could also improve the milk nutritional quality.

  18. Composition and fatty acid distribution of bovine milk phospholipids from processed milk products.

    Science.gov (United States)

    Gallier, Sophie; Gragson, Derek; Cabral, Charles; Jiménez-Flores, Rafael; Everett, David W

    2010-10-13

    The aim of this work was to assess the accuracy of different extraction methods of phospholipids and to measure the effect that processing has on phospholipid composition. Four methods of extracting phospholipids from buttermilk powder were compared to optimize recovery of sphingomyelin. Using the optimal method, the phospholipid profile of four dairy products (raw milk, raw cream, homogenized and pasteurized milk, and buttermilk powder) was determined. A total lipid extraction by the Folch method followed by a solid-phase extraction using the Bitman method was the most efficient technique to recover milk sphingomyelin. Milk processing (churning, centrifuging, homogenization, spray-drying) affected the profile of milk phospholipids, leading to a loss of sphingomyelin and phosphatidylcholine after centrifugation for cream separation. A corresponding decrease in the saturation content of the raw cream phospholipids and a loss of phosphatidylethanolamine after spray-drying to produce buttermilk powder were also observed.

  19. Genetic and environmental variation in bovine milk infrared spectra.

    Science.gov (United States)

    Wang, Qiuyu; Hulzebosch, Alex; Bovenhuis, Henk

    2016-08-01

    Fourier transform infrared (FTIR) spectroscopy is widely used to determine milk composition. In this study, 1,060 milk infrared wavenumbers ranging from 925 to 5,008cm(-1) of 1,748 Holstein Friesian cows on 371 herds in the Netherlands were available. The extent to which infrared wavenumbers are affected by genetic and environmental factors was investigated. Inter-herd heritabilities of 1,060 infrared wavenumbers ranged from 0 to 0.63, indicating that the genetic background of infrared wavenumbers differs considerably. The majority of the wavenumbers have moderate to high inter-herd heritabilities ranging from 0.20 to 0.60. The diacylglycerol O-acyltransferase 1 (DGAT1), stearoyl-CoA desaturase (SCD1), κ-casein (CSN3), and β-lactoglobulin (LGB) polymorphisms are known to have a large effect on milk composition, and therefore we studied the effects of these polymorphisms on infrared wavenumbers. The DGAT1 polymorphism had highly significant effects on many wavenumbers. In contrast, the SCD1 polymorphism did not significantly affect any of the wavenumbers. The SCD1 is known to have a strong effect on the content of C10:1, C12:1, C14:1, and C16:1 fatty acids. Therefore, these results suggest that FTIR spectra contain little direct information on these monounsaturated fatty acids. The CSN3 and LGB polymorphisms had significant effects on a few wavenumbers. Differences between herds explained 10 to 25% of the total variance for most wavenumbers. This suggests that the wavenumbers of milk FTIR spectra are indicative for differences in feeding and management between herds. The wavenumbers between 1,619 and 1,674cm(-1) and between 3,073 and 3,667cm(-1) are strongly influenced by water absorption and usually excluded when setting up prediction equations. However, we found that some of the wavenumbers in the water absorption region are affected by the DGAT1 polymorphism and lactation stage. This suggests that these wavenumbers contain useful information regarding milk

  20. Similar efficacy of human banked milk and bovine colostrum to decrease incidence of necrotizing enterocolitis in preterm piglets

    DEFF Research Database (Denmark)

    Jensen, Michael L.; Sangild, Per Torp; Lykke, Mikkel;

    2013-01-01

    Preterm birth and formula feeding predispose to necrotizing enterocolitis (NEC) in infants. As mother's milk is often absent following preterm delivery, infant formula (IF) and human donor milk (HM) are frequently used as alternatives. We have previously shown that porcine and bovine colostrum (BC...... to IF in stimulating gut structure, function, and NEC resistance in preterm piglets. BC may be a relevant alternative to HM when mother's milk is unavailable during the first week after preterm birth....

  1. Comparative Analysis of the miRNome of Bovine Milk Fat, Whey and Cells.

    Science.gov (United States)

    Li, Ran; Dudemaine, Pier-Luc; Zhao, Xin; Lei, Chuzhao; Ibeagha-Awemu, Eveline Mengwi

    2016-01-01

    alternative non-invasive source of RNA in assessing miRNA activities in bovine mammary gland. Predicted target genes (1802) of 14 highly expressed miRNAs in milk fractions were enriched in fundamental cellular functions, infection, organ and tissue development. Furthermore, some miRNAs were highly enriched (FDR milk whey (3), cells (11) and mammary gland tissue (14) suggesting specific regulatory functions in the various fractions. In conclusion, we have obtained a comprehensive miRNA profile of the different milk fractions using high throughput sequencing. Our comparative analysis showed that miRNAs from milk fat accurately portrayed the miRNome of mammary gland tissue. Functional annotation of the top expressed miRNAs in milk confirmed their critical regulatory roles in mammary gland functions and potentially to milk recipients.

  2. Synthesis and bioevaluation of 2-phenyl-4-methyl-1,3-selenazole-5-carboxylic acids as potent xanthine oxidase inhibitors.

    Science.gov (United States)

    Guan, Qi; Cheng, Zengjin; Ma, Xiaoxue; Wang, Lijie; Feng, Dongjie; Cui, Yuanhang; Bao, Kai; Wu, Lan; Zhang, Weige

    2014-10-06

    A series of 2-phenyl-4-methyl-1,3-selenazole-5-carboxylic acid derivatives (8a-f, 9a-m) were synthesized and evaluated for inhibitory activity against xanthine oxidase in vitro. Structure-activity relationship analyses have also been presented. Most of the target compounds exhibited potency levels in the nanomolar range. Compound 9e emerged as the most potent xanthine oxidase inhibitor (IC50 = 5.5 nM) in comparison to febuxostat (IC50 = 18.6 nM). Steady-state kinetics measurements with the bovine milk enzyme indicated a mixed type inhibition with Ki and Ki' values of 0.9 and 2.3 nM, respectively. A molecular modeling study on compounds 9e was performed to gain an insight into its binding mode with xanthine oxidase, and to provide the basis for further structure-guided design of new non-purine xanthine oxidase inhibitors related with 2-phenyl-4-methyl-1,3-selenazole-5-carboxylic acid scaffold.

  3. Development of a TIRF-based biosensor for sensitive detection of progesterone in bovine milk.

    Science.gov (United States)

    Käppel, Nina D; Pröll, Florian; Gauglitz, Guenter

    2007-04-15

    A total internal reflectance fluorescence (TIRF)-based biosensor for progesterone in bovine milk was developed and tested by measuring the progesterone level in daily milk samples for 25 days, covering a whole estrus cycle. The detection is based on total internal reflectance fluorescence. The assay has been designed as a binding-inhibition test with a progesterone derivative covalently immobilized on the sensor surface and a monoclonal anti-progesterone antibody as biological recognition element. First an existing progesterone assay was optimized by reducing the assay time per measurement, resulting in an assay time of about 5 min and reaching a limit of detection (LOD) of 0.04 ng mL(-1) and a quantification limit (LOQ) of 0.34 ng mL(-1). After calibration the assay was tested by measuring the progesterone level in daily milk samples over several weeks. An estrus cycle of a cow could be measured. As results become available within minutes without any preparation or pre-concentration of the milk samples the fully automated TIRF-based biosensor for progesterone can be used in-line in the milking parlor and thus could be an important tool for reproductive management of dairy cattle detecting heat and predicting pregnancy, which are critical parameters in milk production.

  4. Antibacterial susceptibility of bovine-mastitis pathogens tested directly in milk from infected quarters.

    Science.gov (United States)

    Louhi-Lehtiö, M; Sandholm, M; Myllys, V; Honkanen-Buzalski, T

    1994-04-01

    Antibacterial susceptibilities of bovine-mastitis pathogens were analysed directly in 57 mastitic milk samples without inoculation with exogenous organisms. Aseptically collected milk was mixed with serial dilutions of antibacterials and the growth was observed using 2,3,5-triphenyltetrazolium chloride (TTC) reduction the following day. The results were compared with those obtained by using calibrated bacterial inocula in turbidimetric minimum-inhibitory-concentration (MIC) determination in broth cultures, and in TTC-broth culture-test and TTC-normal milk-test. The results of different methods all correlated positively when the entire data was used. However, taking the direct test in mastitic milk as the 'true' result, the total discrepancies varied from 34.7% to 48.8%. Antibacterial activities of the trimethoprim-sulphadoxine combination, and of spiramycin and ampicillin, decreased significantly when nutrient broth was replaced by milk as the test medium. The efficacy of trimethoprim-sulphadoxine as an antibacterial agent was also dependent on the source of milk.

  5. Study of the protein-bound fraction of calcium, iron, magnesium and zinc in bovine milk

    Science.gov (United States)

    Silva, Fernando V.; Lopes, Gisele S.; Nóbrega, Joaquim A.; Souza, Gilberto B.; Nogueira, Ana Rita A.

    2001-10-01

    Two approaches were used to study the interaction of Ca, Fe, Mg and Zn with bovine milk proteins by inductively coupled plasma optical emission spectrometry (ICPOES). Selective separations in bovine milk samples were accomplished employing an acid protein precipitation using 100 g l -1 trichloroacetic acid (TCA), and an enzymatic protein hydrolysis using 50 g l -1 pepsin (PEP) solution, respectively. The results were compared with total mineral contents determined after microwave-assisted acid digestion. The results obtained by enzymatic and acid precipitation evidenced the different interaction forms of Ca, Fe, Mg and Zn in the system formed by milk components. Iron was not solubilized by the TCA treatment, but was recovered completely after the enzymatic treatment. Quantitative recoveries of Ca, Mg and Zn were obtained using both approaches, showing that these analytes were bound to milk compounds affected by either treatment. Calcium, Mg and Zn are mainly associated with colloidal calcium phosphate and Fe is bound to the backbone of the casein polypeptide chain, cleaved by pepsin enzyme. The proposed approaches could be used to assess the complexity of these chemical interactions.

  6. Electrophoretic and immunological properties of folate-binding protein isolated from bovine milk

    Energy Technology Data Exchange (ETDEWEB)

    Iwai, Kazuo; Tani, Masako; Fushiki, Tohru (Kyoto Univ. (Japan). Faculty of Agriculture)

    1983-07-01

    Changes of the folate-binding protein (FBP) concentration in bovine milk after parturition were investigated. The FBP was highly purified from mature milk by affinity chromatography. The purified FBP showed a single protein band in polyacrylamide gel electrophoresis and was immunologically homogenous in double immunodiffusion. However, in two-dimensional gel electrophoresis, the FBP was separated into several spots in isoelectric focusing in the first dimension, and each spot also showed two molecular weights in SDS-gel electrophoresis in the second dimension. But these FBP molecules were immunologically identical with each other. The neuraminidase treatment obviously diminished the number of isoelectric points of the FBP. Thus, the variety of FBP molecules was at least partially due to the variability of the sialic acid content in the carbohydrate moieties. Moreover, the milk FBP showed species-specificity among mammals immunologically as well as physicochemically.

  7. Organochlorine pesticide residues in bovine milk from organic farms in Chiapas, Mexico.

    Science.gov (United States)

    Gutiérrez, Rey; Ruíz, Jorge Luis; Ortiz, Rutilio; Vega, Salvador; Schettino, Beatriz; Yamazaki, Alberto; de Lourdes Ramírez, María

    2012-10-01

    Thirty six samples of bovine milk were collected from Chiapas State, Mexico between January 2011 and December 2011 with the intention of identifying and quantifying organochlorine pesticide residues in organic farms. The analyses were done using gas chromatography with an electron capture detector (Ni(63)). In general the values found in raw milk were lower than the permissible limit proposed by FAO/WHO/Codex Alimentarius 2006. Average concentrations for alpha + beta HCH were 3.62 ng/g, gamma HCH 0.34 ng/g, heptachlor + epoxide 0.67 ng/g, DDT and isomers 1.53 ng/g, aldrin + dieldrin 0.77 ng/g, and endrin 0.66 ng/g (only present in samples from farm 2). The organic milk from Chiapas has shown low concentrations of pesticide residues in recent years and satisfies international and national regulations for commercialization.

  8. Time resolved bovine host reponse to virulence factors mapped in milk by selected reaction monitoring

    DEFF Research Database (Denmark)

    Bislev, Stine Lønnerup; Kusebauch, Ulrike; Codrea, Marius Cosmin

    TIME RESOLVED BOVINE HOST RESPONSE TO VIRULENCE FACTORS, MAPPED IN MILK BY SELECTED REACTION MONITORING S.L. Bislev1, U. Kusebauch2, M.C. Codrea1, R. Moritz2, C.M. Røntved1, E. Bendixen1 1 Department of Animal Science, Faculty of Science and Technology, Aarhus University, Tjele, Denmark; 2...... Institute for Systems Biology, Seattle, Washington, USA Mastitis is beyond doubt the largest health problem in modern milk production. Many different pathogens can cause infections in the mammary gland, and give rise to severe toll on animal welfare, economic gain as well as on excessive use of antibiotics...... in food production. Rapid diagnostic methods are still not available, and particularly pathogen-specific biomarkers would be highly valuable, as these may allow correct antibiotic treatment to be applied shortly after an udder infection has been observed. Moreover, with automatic milking systems and on...

  9. Comparative evaluation of various solid phases for the development of coated tube assays for the estimation of progesterone in human serum, bovine serum and bovine milk

    Energy Technology Data Exchange (ETDEWEB)

    Karir, Tarveen [Board of Radiation and Isotope Technology (BRIT), BARC Vashi Complex, Department of Atomic Energy (DAE), Navi Mumbai 400 705 (India); Samuel, Grace [Board of Radiation and Isotope Technology (BRIT), BARC Vashi Complex, Department of Atomic Energy (DAE), Navi Mumbai 400 705 (India)], E-mail: grace_samuel1955@rediffmail.com; Sivaprasad, N.; Venkatesh, Meera [Board of Radiation and Isotope Technology (BRIT), BARC Vashi Complex, Department of Atomic Energy (DAE), Navi Mumbai 400 705 (India)

    2009-11-15

    Immobilization of progesterone antibody using three polystyrene surfaces and two progesterone radiotracers for use in the development of a coated tube assay for the evaluation of progesterone levels in human serum, bovine serum and bovine milk was studied. The selection of the solid phase and the tracers were based on the maximum binding, non-specific binding, sensitivity and percentage recovery. Amongst the polystyrene tubes studied, streptavidin coated tubes showed the acceptable assay features such as low non-specific binding (0.5-1.0%), adequate sensitivity (0.13-0.16 ng/ml) and recovery (85-115%) for all the three sample matrices, human serum, bovine serum and bovine milk.

  10. Purification and separation of multiple forms of lactophorin from bovine milk whey and their immunological and electrophoretic properties.

    Science.gov (United States)

    Kanno, C

    1989-04-01

    Lactophorin is designated as a glycoprotein, which is present in bovine milk whey and reacts to the antiserum of the soluble glycoprotein of bovine milk fat globule membrane. The lactophorin was purified by DEAE-cellulose (pH 7.7), Sephadex G-100, and then Bio Gel A-15m from the component-3 fraction of the proteose-peptone fraction of bovine milk whey. The purified lactophorin was separated into seven components by DEAE-cellulose chromatography at pH 8.6. The seven components (LP-1 to -7) of lactophorin were almost homogeneous, but the respective bands were somewhat broad and varied in mobilities on disc electrophoresis. The seven lactophorin components fused completely to the antisoluble glycoprotein of milk fat globule membrane on double immunodiffusion but showed different mobilities of precipitation lines on immunoelectrophoresis. The results indicated that lactophorin consisted of multiple forms but had a common set of antigenic determinant groups against anti-soluble glycoprotein.

  11. The glycemic, insulinemic and plasma amino acid responses to equi-carbohydrate milk meals, a pilot- study of bovine and human milk

    Directory of Open Access Journals (Sweden)

    Gunnerud Ulrika

    2012-10-01

    Full Text Available Abstract Background Dairy proteins, in particular the whey fraction, exert insulinogenic properties and facilitate glycemic regulation through a mechanism involving elevation of certain plasma amino acids, and stimulation of incretins. Human milk is rich in whey protein and has not been investigated in this respect. Method Nine healthy volunteers were served test meals consisting of human milk, bovine milk, reconstituted bovine whey- or casein protein in random order. All test meals contributed with 25g intrinsic or added lactose, and a white wheat bread (WWB meal was used as reference, providing 25g starch. Post-prandial levels in plasma of glucose, insulin, incretins and amino acids were investigated at time intervals for up to 2 h. Results All test meals elicited lower postprandial blood glucose responses, expressed as iAUC 0–120 min compared with the WWB (P  Conclusion This study shows that the glycemic response was significantly lower following all milk/milk protein based test meals, in comparison with WWB. The effect appears to originate from the protein fraction and early phase plasma amino acids and incretins were involved in the insulin secretion. Despite its lower protein content, the human milk was a potent GLP-1 secretagogue and showed insulinogenic properties similar to that seen with reconstituted bovine whey-protein, possibly due to the comparatively high proportion of whey in human milk.

  12. Carryover of bovine leukemia virus antibodies in samples from shared milk meters.

    Science.gov (United States)

    Nekouei, O A; Sanchez, J; Keefe, G P

    2015-08-01

    Screening for infectious diseases of cattle using milk from the dairy herd improvement (DHI) sampling process is very convenient. However, when samples from shared milk meters are used, carryover of antibodies or other diagnostic targets can complicate the interpretation of the diagnostic test results for diseases, including bovine leukosis. The objectives of this study were (1) to assess the potential for carryover of antibodies against bovine leukemia virus (BLV) in milk samples obtained from shared meters, and (2) to determine if adjustment of the diagnostic test cut-off value would improve the test characteristics for meter-collected milk ELISA results. Eight dairy farms were randomly selected from herds with a wide range of BLV prevalence levels in Prince Edward Island, Canada. Within each chosen farm, 2 to 4milk meters were randomly selected. During the routine procedures of DHI sampling, 2 simultaneous milk samples, 1 hand-collected at the beginning of milking (after udder preparation) and the other from the corresponding milk meter, were taken from all lactating cows (n=236) that were milked at the selected meters (n=26). The sequence of cows using each meter was recorded. All samples were tested for BLV antibodies using a commercial indirect ELISA. Antibody carryover potential was assessed in meter-collected samples which were preceded by other cows using the same meters. Applying the hand-collected sample results as our reference standard, a new cut-off was defined for meter-collected samples to optimize the test characteristics. At the standard cut-off value of the diagnostic test, 110 (46.6%) of the hand-collected and 136 (57.6%) of the meter-collected samples were positive. For low-titer cows (e.g., true negatives), the likelihood of antibody carryover significantly increased as the titer of preceding cows increased, whereas this change was not substantial for high-titer cows. The odds of obtaining false diagnoses in meter-positive samples became

  13. Comparison of the human and bovine milk N-glycome via high-performance microfluidic chip liquid chromatography and tandem mass spectrometry.

    Science.gov (United States)

    Nwosu, Charles C; Aldredge, Danielle L; Lee, Hyeyoung; Lerno, Larry A; Zivkovic, Angela M; German, J Bruce; Lebrilla, Carlito B

    2012-05-04

    The isolation of whey proteins from human and bovine milks followed by profiling of their entire N-glycan repertoire is described. Whey proteins resulting from centrifugation and ethanol precipitation of milk were treated with PNGase F to release protein-bound N-glycans. Once released, N-glycans were analyzed via nanoflow liquid chromatography coupled with quadrupole time-of-flight mass spectrometry following chromatographic separation on a porous graphitized carbon chip. In all, 38 N-glycan compositions were observed in the human milk sample while the bovine milk sample revealed 51 N-glycan compositions. These numbers translate to over a hundred compounds when isomers are considered and point to the complexity of the mixture. High mannose, neutral, and sialylated complex/hybrid glycans were observed in both milk sources. Although NeuAc sialylation was observed in both milk samples, the NeuGc residue was only observed in bovine milk and marks a major difference between human and bovine milks. To the best of our knowledge, this study is the first MS based confirmation of NeuGc in milk protein bound glycans as well as the first comprehensive N-glycan profile of bovine milk proteins. Tandem MS was necessary for resolving complications presented by the fact that (NeuGc:Fuc) corresponds to the exact mass of (NeuAc:Hex). Comparison of the relative distribution of the different glycan types in both milk sources was possible via their abundances. While the human milk analysis revealed a 6% high mannose, 57% sialylation, and 75% fucosylation distribution, a 10% high mannose, 68% sialylation, and 31% fucosylation distribution was observed in the bovine milk analysis. Comparison with the free milk oligosaccharides yielded low sialylation and high fucosylation in human, while high sialylation and low fucosylation are found in bovine. The results suggest that high fucosylation is a general trait in human, while high sialylation and low fucosylation are general features of

  14. Effects of phenylalanine and threonine oligopeptides on milk protein synthesis in cultured bovine mammary epithelial cells.

    Science.gov (United States)

    Zhou, M M; Wu, Y M; Liu, H Y; Liu, J X

    2015-04-01

    This study was conducted to investigate the effects of phenylalanine (Phe) and threonine (Thr) oligopeptides on αs1 casein gene expression and milk protein synthesis in bovine mammary epithelial cells. Primary mammary epithelial cells were obtained from Holstein dairy cows and incubated in Dulbecco's modified Eagle's medium-F12 medium (DMEM/F12) containing lactogenic hormones (prolactin and glucocorticoids). Free Phe (117 μg/ml) was substituted partly with peptide-bound Phe (phenylalanylphenylalanine, phenylalanyl threonine, threonyl-phenylalanyl-phenylalanine) in the experimental media. After incubation with experimental medium, cells were collected for gene expression analysis and medium was collected for milk protein or amino acid determination. The results showed that peptide-bound Phe at 10% (11.7 μg/ml) significantly enhanced αs1 casein gene expression and milk protein synthesis as compared with equivalent amount of free Phe. When 10% Phe was replaced by phenylalanylphenylalanine, the disappearance of most essential amino acids increased significantly, and gene expression of peptide transporter 2 and some amino acid transporters was significantly enhanced. These results indicate that the Phe and Thr oligopeptides are important for milk protein synthesis, and peptide-bound amino acids could be utilised more efficiently in milk protein synthesis than the equivalent amount of free amino acids.

  15. Identification of flunixin glucuronide and depletion of flunixin and its marker residue in bovine milk.

    Science.gov (United States)

    Jedziniak, P; Olejnik, M; Szprengier-Juszkiewicz, T; Smulski, S; Kaczmarowski, M; Żmudzki, J

    2013-12-01

    Residues of flunixin [and its marker residue 5-hydroxyflunixin (5OHFLU)] were determined in milk from cows that intravenously received therapeutic doses of the drug. The samples were collected during each milking (every 12 h) for six consecutive days, and concentrations of flunixin and its metabolites were determined by the method with and without enzymatic hydrolysis (beta-glucuronidase). The highest flunixin concentration in milk was observed 12 h after dosing (2.4 ± 1.42 μg/kg, mean ± SD). Flunixin concentrations in the samples determined with enzymatic hydrolysis were significantly higher (P flunixin glucuronide to the milk. Additionally, unambiguous identification of flunixin glucuronide in the bovine milk was performed with linear ion-trap mass spectrometry. The 5OHFLU concentrations analyzed without enzymatic hydrolysis (22.3 ± 16.04 μg/kg) were similar to this obtained with enzymatic hydrolysis. Flunixin and 5OHFLU concentrations dropped below the limits of detection at 48 h after last dosing.

  16. Proteome analysis of functionally differentiated bovine (Bos indicus) mammary epithelial cells isolated from milk

    KAUST Repository

    Janjanam, Jagadeesh

    2013-10-01

    Mammary gland is made up of a branching network of ducts that end in alveoli. Terminally differentiated mammary epithelial cells (MECs) constitute the innermost layer of aveoli. They are milk-secreting cuboidal cells that secrete milk proteins during lactation. Little is known about the expression profile of proteins in the metabolically active MECs during lactation or their functional role in the lactation process. In the present investigation, we have reported the proteome map of MECs in lactating cows using 2DE MALDI-TOF/TOF MS and 1D-Gel-LC-MS/MS. MECs were isolated from milk using immunomagnetic beads and confirmed by RT-PCR and Western blotting. The 1D-Gel-LC-MS/MS and 2DE-MS/MS based approaches led to identification of 431 and 134 proteins, respectively, with a total of 497 unique proteins. Proteins identified in this study were clustered into functional groups using bioinformatics tools. Pathway analysis of the identified proteins revealed 28 pathways (p < 0.05) providing evidence for involvement of various proteins in lactation function. This study further provides experimental evidence for the presence of many proteins that have been predicted in annotated bovine genome. The data generated further provide a set of bovine MEC-specific proteins that will help the researchers to understand the molecular events taking place during lactation. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Screening and determination of sulphonamide residues in bovine milk samples using a flow injection system.

    Science.gov (United States)

    Fernandes, Flávio Cesar Bedatty; Silva, Aline Santana; Rufino, José Luiz; Pezza, Helena Redigolo; Pezza, Leonardo

    2015-01-01

    A new, simple, rapid and sensitive flow injection spectrophotometric method was developed for the screening and determination of sulphonamides in bovine milk samples. The method is based on the condensation of sulphathiazole, sulphamethazine, and sulphadimethoxine with p-dimethylaminobenzaldehyde (p-DAB) in acid medium, in the presence of sodium dodecyl sulphate (SDS), producing a yellow compound (λmax=465 nm). Optimisation of the experimental parameters was performed using a multivariate methodology. The linear range was 90-500 μg/L and the limits of detection and quantification were in the ranges 25-29 μg/L and 84-88 μg/L, respectively. The procedure was applied for the determination of sulphonamide antibiotics in bovine milk samples submitted to a prior extraction procedure based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) methodology. Recoveries of 60.5-70.5% were achieved for milk samples spiked with 0.09, 0.1, 0.2, and 0.3 μg/g of each sulphonamide.

  18. Determination of steroid hormones in bovine milk by LC-MS/MS and their levels in Swiss Holstein cow milk.

    Science.gov (United States)

    Goyon, Alexandre; Cai, Julia Zhenzhen; Kraehenbuehl, Karin; Hartmann, Christoph; Shao, Bing; Mottier, Pascal

    2016-05-01

    Synthetic and natural steroid hormones have attracted some attention in recent years as endocrine active substances (EAS) that interact or interfere with the endocrine system. Endogenous hormones occur naturally in food of animal origin, among which bovine milk represents an important source. This study was conducted to determine the occurrence of steroid hormones (oestrogens, androgens, progestogens and glucocorticoids) in cow's milk samples from three farms in Switzerland. An isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of 12 hormones in milk. Some hormonal levels from individual cows showed large variations. The average levels of the hormones analysed (17α-estradiol = 31 ng kg(-)(1), 17β-estradiol = 6 ng kg(-)(1), estrone = 159 ng kg(-)(1), 4-androstenedione = 684 ng kg(-)(1), progesterone = 15486 ng kg(-)(1), 17-hydroxyprogesterone = 214 ng kg(-)(1), cortisone = 112 ng kg(-)(1), and cortisol = 235 ng kg(-)(1)) were comparable with literature data. Estriol, testosterone and androstenediols were not detected at their respective limit of quantification. No significant differences of hormonal content among milk from cows at different lactation/calving numbers were evidenced, except for progesterone and 4-androstenedione. Due to confounding parameters linked to the physiological stage of the animal, like pregnancy and gestational stage (pregnancy trimester), the causal correlation between the variation of the levels for these two hormones and the lactation/calving number could not be unambiguously demonstrated.

  19. Selective extraction and determination of fluoroquinolones in bovine milk samples with montmorillonite magnetic molecularly imprinted polymers and capillary electrophoresis.

    Science.gov (United States)

    Wang, Hongwu; Liu, Yanqing; Wei, Shoulian; Yao, Su; Zhang, Jiali; Huang, Huichang

    2016-01-01

    A sensitive and selective method for separating fluoroquinolones (FQs) from bovine milk samples was successfully developed using montmorillonite magnetic molecularly imprinted polymers (MMMIPs) as adsorbents. MMMIPs were prepared using montmorillonite as carrier, fleroxacin (FLE) as template molecule, and Fe3O4 magnetite as magnetic component. MMMIPs possessed high adsorption capacity of 46.3 mg g(-1) for FLE. A rapid and convenient magnetic solid-phase extraction procedure coupled with capillary electrophoresis was established with MMMIPs as adsorbents for simultaneous and selective extraction of four FQs in bovine milk samples. Limits of detection ranged between 12.9 and 18.8 μg L(-1), and the RSDs were between 1.8% and 8.6%. The proposed method was successfully applied to spike bovine milk samples with recoveries of 92.7%-108.6%.

  20. The Biodiversity of the Microbiota Producing Heat-Resistant Enzymes Responsible for Spoilage in Processed Bovine Milk and Dairy Products.

    Science.gov (United States)

    Machado, Solimar G; Baglinière, François; Marchand, Sophie; Van Coillie, Els; Vanetti, Maria C D; De Block, Jan; Heyndrickx, Marc

    2017-01-01

    Raw bovine milk is highly nutritious as well as pH-neutral, providing the ideal conditions for microbial growth. The microbiota of raw milk is diverse and originates from several sources of contamination including the external udder surface, milking equipment, air, water, feed, grass, feces, and soil. Many bacterial and fungal species can be found in raw milk. The autochthonous microbiota of raw milk immediately after milking generally comprises lactic acid bacteria such as Lactococcus, Lactobacillus, Streptococcus, and Leuconostoc species, which are technologically important for the dairy industry, although they do occasionally cause spoilage of dairy products. Differences in milking practices and storage conditions on each continent, country and region result in variable microbial population structures in raw milk. Raw milk is usually stored at cold temperatures, e.g., about 4°C before processing to reduce the growth of most bacteria. However, psychrotrophic bacteria can proliferate and contribute to spoilage of ultra-high temperature (UHT) treated and sterilized milk and other dairy products with a long shelf life due to their ability to produce extracellular heat resistant enzymes such as peptidases and lipases. Worldwide, species of Pseudomonas, with the ability to produce these spoilage enzymes, are the most common contaminants isolated from cold raw milk although other genera such as Serratia are also reported as important milk spoilers, while for others more research is needed on the heat resistance of the spoilage enzymes produced. The residual activity of extracellular enzymes after high heat treatment may lead to technological problems (off flavors, physico-chemical instability) during the shelf life of milk and dairy products. This review covers the contamination patterns of cold raw milk in several parts of the world, the growth potential of psychrotrophic bacteria, their ability to produce extracellular heat-resistant enzymes and the consequences for

  1. The Biodiversity of the Microbiota Producing Heat-Resistant Enzymes Responsible for Spoilage in Processed Bovine Milk and Dairy Products

    Science.gov (United States)

    Machado, Solimar G.; Baglinière, François; Marchand, Sophie; Van Coillie, Els; Vanetti, Maria C. D.; De Block, Jan; Heyndrickx, Marc

    2017-01-01

    Raw bovine milk is highly nutritious as well as pH-neutral, providing the ideal conditions for microbial growth. The microbiota of raw milk is diverse and originates from several sources of contamination including the external udder surface, milking equipment, air, water, feed, grass, feces, and soil. Many bacterial and fungal species can be found in raw milk. The autochthonous microbiota of raw milk immediately after milking generally comprises lactic acid bacteria such as Lactococcus, Lactobacillus, Streptococcus, and Leuconostoc species, which are technologically important for the dairy industry, although they do occasionally cause spoilage of dairy products. Differences in milking practices and storage conditions on each continent, country and region result in variable microbial population structures in raw milk. Raw milk is usually stored at cold temperatures, e.g., about 4°C before processing to reduce the growth of most bacteria. However, psychrotrophic bacteria can proliferate and contribute to spoilage of ultra-high temperature (UHT) treated and sterilized milk and other dairy products with a long shelf life due to their ability to produce extracellular heat resistant enzymes such as peptidases and lipases. Worldwide, species of Pseudomonas, with the ability to produce these spoilage enzymes, are the most common contaminants isolated from cold raw milk although other genera such as Serratia are also reported as important milk spoilers, while for others more research is needed on the heat resistance of the spoilage enzymes produced. The residual activity of extracellular enzymes after high heat treatment may lead to technological problems (off flavors, physico-chemical instability) during the shelf life of milk and dairy products. This review covers the contamination patterns of cold raw milk in several parts of the world, the growth potential of psychrotrophic bacteria, their ability to produce extracellular heat-resistant enzymes and the consequences for

  2. The Influence of Bovine Milk High or Low in Isoflavones on Hepatic Gene Expression in Mice

    Directory of Open Access Journals (Sweden)

    Mette T. Skaanild

    2010-01-01

    Full Text Available Isoflavones have generated much attention due to their potential positive effects in various diseases. Phytoestrogens especially equol can be found in bovine milk, as feed ration for dairy cows is comprised of plants containing phytoestrogens. The aim of this study was to analyze the changes in hepatic gene expression after dietary intake of milk high and low in isoflavones. In addition to pelleted feed female NMRI mice were offered water, water added either 17-estradiol, equol, Tween 80, and milk high and low in isoflavone content for a week. Gene expression was analyzed using an array qPCR kit. It was revealed that Tween 80 and 17-estradiol upregulated both phase I and phase II genes to the same extent whereas equol alone, high and low isoflavone milk did not alter the expression of phase I genes but decreased the expression of phase II genes. This study shows that dietary isoflavones can regulate the transcription of especially phase II liver enzymes which potentially could give rise to an increase in reactive oxygen metabolites that may contribute to the development of cancer.

  3. Effect of forage on the content of phyto-oestrogens in bovine milk

    DEFF Research Database (Denmark)

    Andersen, Charlotte; Weisbjerg, Martin Riis; Hansen-Møller, Jens

    2009-01-01

    ) affects the concentration of phyto-oestrogens in bovine milk. Sixteen Danish Holstein cows were assigned to a 4 × 4 latin square design with four cows per treatment per period of 3 weeks. The four treatment diets were lucerne silage (LS), 2/3 lucerne silage and 1/3 maize silage (2/3LS), 1/3 lucerne silage...... and 2/3 maize silage (1/3LS) and GCS. Milk was collected at the end of each period and feed samples on day 6, 13 and 20 in each experimental period. Milk and pooled feed samples were analysed for the concentration of isoflavones, coumestans and lignans. The content of isoflavones was higher......Phyto-oestrogens are believed to have a range of beneficial effects on predominant Western diseases. A few studies on phyto-oestrogens in milk exist and show that the composition can be affected by feeding. Therefore, the aim was to study how feeding of lucerne and grass/clover silages (GCSs...

  4. INFLUENCE OF EUROPEAN FUNDS ON THE SECTOR OF BOVINE MILK AND MEAT IN ROMANIA IN THE PERIOD 2007-2013

    Directory of Open Access Journals (Sweden)

    Andrei-Marius SANDU

    2014-12-01

    Full Text Available This study aims to analyze the bovine meat and milk sector for the period 2007-2013. In the analyzed period, Romania has benefited from EU funding through the National Rural Development Programme 2007-2013. In this programme there were measures that addressed exclusively to the animal husbandry sector in Romania. This paper presents the results of the analysis on bovine production of meat, milk and livestock in Romania, but also on the price and impact that the European Funds implementation had on them.

  5. Detection of antibiotic residues in bovine milk by a voltammetric electronic tongue system.

    Science.gov (United States)

    Wei, Zhenbo; Wang, Jun

    2011-05-23

    A voltammetric electronic tongue (VE-tongue) was developed to detect antibiotic residues in bovine milk. Six antibiotics (Chloramphenicol, Erythromycin, Kanamycin sulfate, Neomycin sulfate, Streptomycin sulfate and Tetracycline HCl) spiked at four different concentration levels (0.5, 1, 1.5 and 2 maximum residue limits (MRLs)) were classified based on VE-tongue by two pattern recognition methods: principal component analysis (PCA) and discriminant function analysis (DFA). The VE-tongue was composed of five working electrodes (gold, silver, platinum, palladium, and titanium) positioned in a standard three-electrode configuration. The Multi-frequency large amplitude pulse voltammetry (MLAPV) which consisted of four segments (1 Hz, 10 Hz, 100 Hz and 1000 Hz) was applied as potential waveform. The six antibiotics at the MRLs could not be separated from bovine milk completely by PCA, but all the samples were demarcated clearly by DFA. Three regression models: Principal Component Regression Analysis (PCR), Partial Least Squares Regression (PLSR), and Least Squares-Support Vector Machines (LS-SVM) were used for concentrations of antibiotics prediction. All the regression models performed well, and PCR had the most stable results.

  6. Detection of antibiotic residues in bovine milk by a voltammetric electronic tongue system

    Energy Technology Data Exchange (ETDEWEB)

    Wei Zhenbo [Zhejiang University, Department of Bio-Systems Engineering, 268 Kaixuan Road, Hangzhou, Zhejiang 310029 (China); Wang Jun, E-mail: jwang@zju.edu.cn [Zhejiang University, Department of Bio-Systems Engineering, 268 Kaixuan Road, Hangzhou, Zhejiang 310029 (China)

    2011-05-23

    A voltammetric electronic tongue (VE-tongue) was developed to detect antibiotic residues in bovine milk. Six antibiotics (Chloramphenicol, Erythromycin, Kanamycin sulfate, Neomycin sulfate, Streptomycin sulfate and Tetracycline HCl) spiked at four different concentration levels (0.5, 1, 1.5 and 2 maximum residue limits (MRLs)) were classified based on VE-tongue by two pattern recognition methods: principal component analysis (PCA) and discriminant function analysis (DFA). The VE-tongue was composed of five working electrodes (gold, silver, platinum, palladium, and titanium) positioned in a standard three-electrode configuration. The Multi-frequency large amplitude pulse voltammetry (MLAPV) which consisted of four segments (1 Hz, 10 Hz, 100 Hz and 1000 Hz) was applied as potential waveform. The six antibiotics at the MRLs could not be separated from bovine milk completely by PCA, but all the samples were demarcated clearly by DFA. Three regression models: Principal Component Regression Analysis (PCR), Partial Least Squares Regression (PLSR), and Least Squares-Support Vector Machines (LS-SVM) were used for concentrations of antibiotics prediction. All the regression models performed well, and PCR had the most stable results.

  7. Specificity and Effector Functions of Human RSV-Specific IgG from Bovine Milk.

    Directory of Open Access Journals (Sweden)

    Gerco den Hartog

    Full Text Available BACKGROUND: Respiratory syncytial virus (RSV infection is the second most important cause of death in the first year of life, and early RSV infections are associated with the development of asthma. Breastfeeding and serum IgG have been shown to protect against RSV infection. Yet, many infants depend on bovine milk-based nutrition, which at present lacks intact immunoglobulins. OBJECTIVE: To investigate whether IgG purified from bovine milk (bIgG can modulate immune responses against human RSV. METHODS: ELISAs were performed to analyse binding of bIgG to human respiratory pathogens. bIgG or hRSV was coated to plates to assess dose-dependent binding of bIgG to human Fcγ receptors (FcγR or bIgG-mediated binding of myeloid cells to hRSV respectively. S. Epidermidis and RSV were used to test bIgG-mediated binding and internalisation of pathogens by myeloid cells. Finally, the ability of bIgG to neutralise infection of HEp2 cells by hRSV was evaluated. RESULTS: bIgG recognised human RSV, influenza haemagglutinin and Haemophilus influenza. bIgG bound to FcγRII on neutrophils, monocytes and macrophages, but not to FcγRI and FcγRIII, and could bind simultaneously to hRSV and human FcγRII on neutrophils. In addition, human neutrophils and dendritic cells internalised pathogens that were opsonised with bIgG. Finally, bIgG could prevent infection of HEp2 cells by hRSV. CONCLUSIONS: The data presented here show that bIgG binds to hRSV and other human respiratory pathogens and induces effector functions through binding to human FcγRII on phagocytes. Thus bovine IgG may contribute to immune protection against RSV.

  8. COMPARISON OF CHILDREN'S FOLLOW-ON INSTANT POWDERED COW'S MILK FORMULA, BUFFALO MILK FORMULA AND CHICKEN-BASED FORMULA ON ENAMEL MICROHARDNESS OF BOVINE TEETH IN VITRO.

    Science.gov (United States)

    Vongsavan, Kadkao; Rirattanapong, Praphasri; Surarit, Rudee

    2016-03-01

    Dental caries are a major public health problem worldwide. The aim of this study was to compare the effects of children's follow-on instant powdered cow's milk formula, buffalo milk formula and a chicken-based formula on microhardness of bovine enamel with artificial caries-like lesions. Forty bovine teeth were each placed in acrylic blocks and the enamel surfaces were polished to create flat 5 x 5 millimeter surfaces. The teeth surfaces were then demineralized using 0.1M lactic acid (pH 4.5) to achieve an enamel microhardness of 35-65 Vickers Hardness Numbers (VHN). All specimens were then randomly allocated into one of 4 groups (n=10/group). For remineralization, each group was soaked in a different kind of milk formula for 2 hours at 37°C except group 1 which was a negative control (artificial saliva) group. Group 2 was soaked in Murrah™ buffalo milk formula (a positive control ), group 3 in S-26-Promil-Gold™ (cow's milk formula) and group 4 in a chicken-based formula (Siriraj Hospital, Mahidol University). The microhardness of the specimens was then measured again. Data were analyzed using a one-way ANOVA and paired t-test with a 95% confidence interval. After exposure to the formula, the mean VHN for each study group was significantly higher (paired t-test, p 0.05). In conclusion, S-26-Promil-Gold™ follow-on cow milk formula, Murrah™ buffalo milk formula and the chicken-based formula all increased bovine enamel microhardness after soaking for 2 hours.

  9. Biological Activities of Extracellular Vesicles and Their Cargos from Bovine and Human Milk in Humans and Implications for Infants.

    Science.gov (United States)

    Zempleni, Janos; Aguilar-Lozano, Ana; Sadri, Mahrou; Sukreet, Sonal; Manca, Sonia; Wu, Di; Zhou, Fang; Mutai, Ezra

    2017-01-01

    Extracellular vesicles (EVs) in milk harbor a variety of compounds, including lipids, proteins, noncoding RNAs, and mRNAs. Among the various classes of EVs, exosomes are of particular interest, because cargo sorting in exosomes is a regulated, nonrandom process and exosomes play essential roles in cell-to-cell communication. Encapsulation in exosomes confers protection against enzymatic and nonenzymatic degradation of cargos and provides a pathway for cellular uptake of cargos by endocytosis of exosomes. Compelling evidence suggests that exosomes in bovine milk are transported by intestinal cells, vascular endothelial cells, and macrophages in human and rodent cell cultures, and bovine-milk exosomes are delivered to peripheral tissues in mice. Evidence also suggests that cargos in bovine-milk exosomes, in particular RNAs, are delivered to circulating immune cells in humans. Some microRNAs and mRNAs in bovine-milk exosomes may regulate the expression of human genes and be translated into protein, respectively. Some exosome cargos are quantitatively minor in the diet compared with endogenous synthesis. However, noncanonical pathways have been identified through which low concentrations of dietary microRNAs may alter gene expression, such as the accumulation of exosomes in the immune cell microenvironment and the binding of microRNAs to Toll-like receptors. Phenotypes observed in infant-feeding studies include higher Mental Developmental Index, Psychomotor Development Index, and Preschool Language Scale-3 scores in breastfed infants than in those fed various formulas. In mice, supplementation with plant-derived MIR-2911 improved the antiviral response compared with controls. Porcine-milk exosomes promote the proliferation of intestinal cells in mice. This article discusses the above-mentioned advances in research concerning milk exosomes and their cargos in human nutrition. Implications for infant nutrition are emphasized, where permitted, but data in infants are

  10. Use of SDS-PAGE in Detecting Bovine Milk Content in Adulterated Xinjiang Non-bovine Milk%SDS-PAGE电泳在鉴别用牛乳掺伪的新疆特种乳中的应用

    Institute of Scientific and Technical Information of China (English)

    阿力木·吾布力; 杨洁; 苏力坦·阿巴百克力

    2016-01-01

    Objective] The aim was to use SDS-PAGE to detect adulterated non-bovine milk and measure bovine milk content and confirm the lowest adulteration ratio.[Method]By using different ratio of(1∶1, 1∶4, 1∶10, 1∶20, 1∶100, 1∶200 and 1∶400(Vbovine milk/Vnon-bo-vine milk) )adulterated non-bovine milk, whey protein was isolated and SDS-PAGE was conducted to locate the indicating protein then use gray analysis to detect adulteration and calculate bovine milk content.[Result] The SDS-PAGE can efficiently separate each animal’s whey protein as well as bovine whey protein from adulterated whey.[Conclusion] The lowest adulterating ratio can be detected with SDS-PAGE as 1∶10 (Vbovine milk/Vnon-bovine milk).%[目的]采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳( SDS-PAGE)鉴别一定比例掺伪的特种乳中所含的牛乳成分及确定最低检测掺伪比例。[方法]按一定的比例(1∶1、1∶4、1∶10、1∶20、1∶100、1∶200和1∶400(V牛乳/V特种乳))掺伪,离心分离乳清蛋白,SDS-PAGE电泳,确定掺伪差异性蛋白并进行灰度分析。[结果] SDS-PAGE可以快速、有效地分离各乳乳清蛋白以及掺伪的牛乳乳清蛋白。[结论] SDS-PAGE鉴别掺伪的最低比例为1∶10(V牛乳/V特种乳)。

  11. Bovine and soybean milk bioactive compounds: Effects on inflammatory response of human intestinal Caco-2 cells.

    Science.gov (United States)

    Calvello, Rosa; Aresta, Antonella; Trapani, Adriana; Zambonin, Carlo; Cianciulli, Antonia; Salvatore, Rosaria; Clodoveo, Maria Lisa; Corbo, Filomena; Franchini, Carlo; Panaro, Maria Antonietta

    2016-11-01

    In this study the effects of commercial bovine and soybean milks and their bioactive compounds, namely genistein, daidzein and equol, on the inflammatory responses induced by lipopolysaccharide (LPS) treatment of human intestinal Caco-2 cells were examined, in terms of nitric oxide (NO) release and inducible nitric oxide synthetase (iNOS) expression. Both milks and their bioactive compounds significantly inhibited, dose-dependently, the expression of iNOS mRNA and protein, resulting in a decreased NO production. The NF-κB activation in LPS-stimulated intestinal cells was also examined. In all cases we observed that cell pre-treatment before LPS activation inhibited the IkB phosphorylation. Accordingly, quantification of bioactive compounds by solid phase microextraction coupled with liquid chromatography has shown that they were absorbed, metabolized and released by Caco-2 cells in culture media. In conclusion, we demonstrated that milks and compounds tested are able to reduce LPS-induced inflammatory responses from intestinal cells, interfering with NF-kB dependent molecular mechanisms.

  12. Tudor-SN Regulates Milk Synthesis and Proliferation of Bovine Mammary Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Jinxia Ao

    2015-12-01

    Full Text Available Tudor staphylococcal nuclease (Tudor-SN is a highly conserved and ubiquitously expressed multifunctional protein, related to multiple and diverse cell type- and species-specific cellular processes. Studies have shown that Tudor-SN is mainly expressed in secretory cells, however knowledge of its role is limited. In our previous work, we found that the protein level of Tudor-SN was upregulated in the nucleus of bovine mammary epithelial cells (BMEC. In this study, we assessed the role of Tudor-SN in milk synthesis and cell proliferation of BMEC. We exploited gene overexpression and silencing methods, and found that Tudor-SN positively regulates milk synthesis and proliferation via Stat5a activation. Both amino acids (methionine and estrogen triggered NFκB1 to bind to the gene promoters of Tudor-SN and Stat5a, and this enhanced the protein level and nuclear localization of Tudor-SN and p-Stat5a. Taken together, these results suggest the key role of Tudor-SN in the transcriptional regulation of milk synthesis and proliferation of BMEC under the stimulation of amino acids and hormones.

  13. Transcobalamin derived from bovine milk stimulates apical uptake of vitamin B12 into human intestinal epithelial cells.

    Science.gov (United States)

    Hine, Brad; Boggs, Irina; Green, Ralph; Miller, Joshua W; Hovey, Russell C; Humphrey, Rex; Wheeler, Thomas T

    2014-11-01

    Intestinal uptake of vitamin B12 (hereafter B12) is impaired in a significant proportion of the human population. This impairment is due to inherited or acquired defects in the expression or function of proteins involved in the binding of diet-derived B12 and its uptake into intestinal cells. Bovine milk is an abundant source of bioavailable B12 wherein it is complexed with transcobalamin. In humans, transcobalamin functions primarily as a circulatory protein, which binds B12 following its absorption and delivers it to peripheral tissues via its cognate receptor, CD320. In the current study, the transcobalamin-B12 complex was purified from cows' milk and its ability to stimulate uptake of B12 into cultured bovine, mouse and human cell lines was assessed. Bovine milk-derived transcobalamin-B12 complex was absorbed by all cell types tested, suggesting that the uptake mechanism is conserved across species. Furthermore, the complex stimulated the uptake of B12 via the apical surface of differentiated Caco-2 human intestinal epithelial cells. These findings suggest the presence of an alternative transcobalamin-mediated uptake pathway for B12 in the human intestine other than that mediated by the gastric glycoprotein, intrinsic factor. Our findings highlight the potential for transcobalamin-B12 complex derived from bovine milk to be used as a natural bioavailable alternative to orally administered free B12 to overcome B12 malabsorption.

  14. Longitudinal metagenomic profiling of bovine milk to assess the impact of intramammary treatment using a third-generation cephalosporin

    NARCIS (Netherlands)

    Ganda, Erika K.; Bisinotto, Rafael S.; Lima, Svetlana F.; Kronauer, Kristina; Decter, Dean H.; Oikonomou, Georgios; Schukken, Ynte H.; Bicalho, Rodrigo C.

    2016-01-01

    Antimicrobial usage in food animals has a direct impact on human health, and approximately 80% of the antibiotics prescribed in the dairy industry are used to treat bovine mastitis. Here we provide a longitudinal description of the changes in the microbiome of milk that are associated with mastitis

  15. Technical note: Identification of Prototheca species from bovine milk samples by PCR-single strand conformation polymorphism.

    Science.gov (United States)

    Cremonesi, P; Pozzi, F; Ricchi, M; Castiglioni, B; Luini, M; Chessa, S

    2012-12-01

    We report the development of a PCR-single strand conformation polymorphism (SSCP) method to identify Prototheca spp. responsible for bovine mastitis: P. zopfii and P. blaschkeae. The method was set up using reference strains belonging to P. zopfii genotype 1, P. zopfii genotype 2, and P. blaschkeae as target species and P. stagnora, and P. ulmea as negative controls. The assay was applied on 50 isolates of Prototheca spp. isolated from bovine mastitic milk or bulk-tank milk samples, and all isolates were identified as P. zopfii genotype 2. We conclude that the described PCR-SSCP approach is accurate, inexpensive, and highly suitable for the identification of P. zopfii genotype 2 on field isolates but also directly on milk, if preceded by a specific DNA extraction method.

  16. Rapid Screening of Bovine Milk Oligosaccharides in a Whey Permeate Product and Domestic Animal Milks by Accurate Mass Database and Tandem Mass Spectral Library.

    Science.gov (United States)

    Lee, Hyeyoung; Cuthbertson, Daniel J; Otter, Don E; Barile, Daniela

    2016-08-17

    A bovine milk oligosaccharide (BMO) library, prepared from cow colostrum, with 34 structures was generated and used to rapidly screen oligosaccharides in domestic animal milks and a whey permeate powder. The novel library was entered into a custom Personal Compound Database and Library (PCDL) and included accurate mass, retention time, and tandem mass spectra. Oligosaccharides in minute-sized samples were separated using nanoliquid chromatography (nanoLC) coupled to a high resolution and sensitive quadrupole-Time of Flight (Q-ToF) MS system. Using the PCDL, 18 oligosaccharides were found in a BMO-enriched product obtained from whey permeate processing. The usefulness of the analytical system and BMO library was further validated using milks from domestic sheep and buffaloes. Through BMO PCDL searching, 15 and 13 oligosaccharides in the BMO library were assigned in sheep and buffalo milks, respectively, thus demonstrating significant overlap between oligosaccharides in bovine (cow and buffalo) and ovine (sheep) milks. This method was shown to be an efficient, reliable, and rapid tool to identify oligosaccharide structures using automated spectral matching.

  17. Effect of bovine ABCG2 polymorphism Y581S SNP on secretion into milk of enterolactone, riboflavin and uric acid.

    Science.gov (United States)

    Otero, J A; Miguel, V; González-Lobato, L; García-Villalba, R; Espín, J C; Prieto, J G; Merino, G; Álvarez, A I

    2016-02-01

    The ATP-binding cassette transporter G2/breast cancer resistance protein (ABCG2/BCRP) is an efflux protein involved in the bioavailability and milk secretion of endogenous and exogenous compounds, actively affecting milk composition. A limited number of physiological substrates have been identified. However, no studies have reported the specific effect of this polymorphism on the secretion into milk of compounds implicated in milk quality such as vitamins or endogenous compounds. The bovine ABCG2 Y581S polymorphism is described as a gain-of-function polymorphism that increases milk secretion and decreases plasma levels of its substrates. This work aims to study the impact of Y581S polymorphism on plasma disposition and milk secretion of compounds such as riboflavin (vitamin B2), enterolactone, a microbiota-derived metabolite from the dietary lignan secoisolariciresinol and uric acid. In vitro transport of these compounds was assessed in MDCK-II cells overexpressing the bovine ABCG2 (WT-bABCG2) and its Y581S variant (Y581S-bABCG2). Plasma and milk levels were obtained from Y/Y homozygous and Y/S heterozygous cows. The results show that riboflavin was more efficiently transported in vitro by the Y581S variant, although no differences were noted in vivo. Both uric acid and enterolactone were substrates in vitro of the bovine ABCG2 variants and were actively secreted into milk with a two-fold increase in the milk/plasma ratio for Y/S with respect to Y/Y cows. The in vitro ABCG2-mediated transport of the drug mitoxantrone, as a model substrate, was inhibited by enterolactone in both variants, suggesting the possible in vivo use of this enterolignan to reduce ABCG2-mediated milk drug transfer in cows. The Y581S variant was inhibited to a lesser extent probably due to its higher transport capacity. All these findings point to a significant role of the ABCG2 Y581S polymorphism in the milk disposition of enterolactone and the endogenous molecules riboflavin and uric acid

  18. Bovine beta-lactoglobulin in human milk from atopic and non-atopic mothers. Relationship to maternal intake of homogenized and unhomogenized milk

    DEFF Research Database (Denmark)

    Høst, A; Husby, S; Hansen, L G;

    1990-01-01

    ). In a cross-over design the atopic and non-atopic mothers alternated their intake of milk between homogenized and unhomogenized milk each week. On day 7, in each week, consecutive milk samples were taken before and 4, 8, 12 and 24 hr after a single ingestion of 500 ml of homogenized or unhomogenized milk....... Detectable amounts of BLG (0.9-150 micrograms/l, median value 4.2 micrograms/l) were measured in 19/20 of the mothers (95%), in 9 of 10 atopic mothers and in all 10 of 10 non-atopic mothers. No correlation was found between the type of milk preparation (homogenized or unhomogenized) and the presence of BLG......Human milk samples (n = 300) were collected during a 3-week period from 10 healthy mothers and from 10 atopic mothers, all with healthy, solely breast-fed infants. The milk samples were analysed by an enzyme-linked immunosorbent assay (ELISA) for the content of bovine beta-lactoglobulin (BLG...

  19. Prediction of individual milk proteins including free amino acids in bovine milk using mid-infrared spectroscopy and their correlations with milk processing characteristics.

    Science.gov (United States)

    McDermott, A; Visentin, G; De Marchi, M; Berry, D P; Fenelon, M A; O'Connor, P M; Kenny, O A; McParland, S

    2016-04-01

    The aim of this study was to evaluate the effectiveness of mid-infrared spectroscopy in predicting milk protein and free amino acid (FAA) composition in bovine milk. Milk samples were collected from 7 Irish research herds and represented cows from a range of breeds, parities, and stages of lactation. Mid-infrared spectral data in the range of 900 to 5,000 cm(-1) were available for 730 milk samples; gold standard methods were used to quantify individual protein fractions and FAA of these samples with a view to predicting these gold standard protein fractions and FAA levels with available mid-infrared spectroscopy data. Separate prediction equations were developed for each trait using partial least squares regression; accuracy of prediction was assessed using both cross validation on a calibration data set (n=400 to 591 samples) and external validation on an independent data set (n=143 to 294 samples). The accuracy of prediction in external validation was the same irrespective of whether undertaken on the entire external validation data set or just within the Holstein-Friesian breed. The strongest coefficient of correlation obtained for protein fractions in external validation was 0.74, 0.69, and 0.67 for total casein, total β-lactoglobulin, and β-casein, respectively. Total proteins (i.e., total casein, total whey, and total lactoglobulin) were predicted with greater accuracy then their respective component traits; prediction accuracy using the infrared spectrum was superior to prediction using just milk protein concentration. Weak to moderate prediction accuracies were observed for FAA. The greatest coefficient of correlation in both cross validation and external validation was for Gly (0.75), indicating a moderate accuracy of prediction. Overall, the FAA prediction models overpredicted the gold standard values. Near-unity correlations existed between total casein and β-casein irrespective of whether the traits were based on the gold standard (0.92) or mid

  20. Determination of 11 quinolones in bovine milk using immunoaffinity stir bar sorptive microextraction and liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Yao, Kai; Zhang, Wei; Yang, Linyan; Gong, Jianfang; Li, Liuan; Jin, Tianming; Li, Cun

    2015-10-15

    A sensitive, selective and reproducible immunoaffinity stir bar sorptive microextraction (SBSME) coupled with liquid chromatography-fluorescence method for determination of 11 quinolones (QNs) in bovine milk was developed and validated. It is first report of a broad-specificity monoclonal antibody to QNs that has been immobilized to glass bar for preparation of a re-usable immunoaffinity stir bar. Analytes were extracted by placing stir bar in milk and shaking on a rotary shaker for 30min at 30rpm, followed by liquid chromatography and fluorescence detection. The newly developed method has limits of detection for each QN from 0.05 to 0.1ng/g with intra-day and inter-day precision ranging from 3.2 to 11.9% and from 5.2 to 12.5%, respectively. This allowed us to quantitatively analyze drugs in bovine milk with the advantage of significantly simplified sample preparation. The proposed method was successfully applied to the bovine milk samples analyses with QNs, demonstrating its rare application in animal food safety analysis.

  1. Bovine kappa-casein gene polymorphism and its association with milk production traits

    Directory of Open Access Journals (Sweden)

    Satyanarayana Rachagani

    2008-12-01

    Full Text Available Point mutations in exon IV of the bovine κ-casein (CSN3 gene determine two allelic variants, A and B. These variants were distinguished by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP analysis in the indigenous Sahiwal and Tharparkar cattle breeds. DNA samples (252 Sahiwal and 56 Tharparkar were analyzed for allelic variants of the CSN3 gene. Polymorphism was detected by digestion of PCR-amplified products with HindIII, HhaI and HaeIII restriction enzymes, followed by separation on 3% agarose gels, and resolved by ethidium bromide staining. Allele A of the κ-casein gene occurred at a higher frequency than allele B, in both Sahiwal and Tharparkar breeds. The genotypic frequencies of AA, AB, and BB in the Sahiwal and Tharparkar breeds were 0.758, 0.230 and 0.012, and 0.0.732, 0.250 and 0.018, respectively. The frequencies of alleles A and B in the Sahiwal and Tharparkar breeds were 0.873 and 0.127, and 0.857 and 0.143, respectively. Genotype BB of the kappa-casein gene had more influence on the monthly milk yield, 305-days milk yield, monthly solids-not-fat (SNF yield, and monthly protein yield, in the Sahiwal cattle.

  2. Caseins from bovine colostrum and milk strongly bind piscidin-1, an antimicrobial peptide from fish.

    Science.gov (United States)

    Kütt, Mary-Liis; Stagsted, Jan

    2014-09-01

    A model system of bovine colostrum and piscidin, a fish-derived antimicrobial peptide, was developed to study potential interactions of antimicrobial peptides in colostrum. We did not detect any antimicrobial activity of colostrum using the radial plate diffusion assay; in fact colostrum completely abrogated activity of added piscidin. This could not be explained by degradation of piscidin by colostrum, which was less than ten percent. We found that colostrum even protected piscidin against degradation by added proteases. We further observed that colostrum and milk rapidly quenched the fluorescence of fluorescein-piscidin but not that of fluorescein. This effect was not seen with BSA and the specific quenching of fluorescein-piscidin by colostrum was saturably inhibited with unlabeled piscidin. Size exclusion chromatography indicated that fluorescein-piscidin bound to casein micelles with no apparent binding to IgG or whey proteins. Further, addition of pure caseins was able to quench fluorescence of fluorescein-piscidin and to inhibit the antimicrobial activity of piscidin. The interaction between caseins and piscidin could be dissociated by guanidine hydrochloride and recovered piscidin had antimicrobial activity against bacteria. Based on our results we propose that caseins could be carriers for antimicrobial peptides in colostrum and milk.

  3. Application of an acid proteinase from Monascus purpureus to reduce antigenicity of bovine milk whey protein.

    Science.gov (United States)

    Lakshman, P L Nilantha; Tachibana, Shinjiro; Toyama, Hirohide; Taira, Toki; Suganuma, Toshihiko; Suntornsuk, Worapot; Yasuda, Masaaki

    2011-09-01

    An acid proteinase from Monascus purpureus No. 3403, MpuAP, was previously purified and some characterized in our laboratory (Agric Biol Chem 48:1637-1639, 1984). However, further information about this enzyme is lacking. In this study, we investigated MpuAP's comprehensive substrate specificity, storage stability, and prospects for reducing antigenicity of whey proteins for application in the food industry. MpuAP hydrolyzed primarily five peptide bonds, Gln(4)-His(5), His(10)-Leu(11), Ala(14)-Leu(15), Gly(23)-Phe(24) and Phe(24)-Phe(25) in the oxidized insulin B-chain. The lyophilized form of the enzyme was well preserved at 30-40°C for 7 days without stabilizers. To investigate the possibility of reducing the antigenicity of the milk whey protein, enzymatic hydrolysates of the whey protein were evaluated by inhibition ELISA. Out of the three main components of whey protein, casein and α-lactalbumin were efficiently degraded by MpuAP. The sequential reaction of MpuAP and trypsin against the whey protein successfully degraded casein, α-lactalbumin and β-lactoglobulin with the highest degree of hydrolysis. As a result, the hydrolysates obtained by using the MpuAP-trypsin combination showed the lowest antigenicity compared with the single application of pepsin, trypsin or pepsin-trypsin combination. Therefore, the overall result suggested that the storage-stable MpuAP and trypsin combination will be a productive approach for making hypoallergic bovine milk whey protein hydrolysates.

  4. Enterotoxin genes in coagulase-negative and coagulase-positive staphylococci isolated from bovine milk.

    Science.gov (United States)

    de Freitas Guimarães, Felipe; Nóbrega, Diego Borin; Richini-Pereira, Virginia Bodelão; Marson, Pâmela Merlo; de Figueiredo Pantoja, José Carlos; Langoni, Helio

    2013-05-01

    The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 10 Brazilian dairy herds and study their capability to produce enterotoxins. Herds were selected based on size and use of milking technology, and farms were visited once during the study. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20 mL) was collected from all CMT-positive quarters. Identification of Staphylococcus spp. was performed using conventional microbiology, and PCR was used to determine the presence of enterotoxin-encoding genes (sea, seb, sec, and sed). Of the 1,318 CMT-positive milk samples, Staphylococcus spp. were isolated from 263 (19.9%). Of these isolates, 135 (51%) were coagulase-positive staphylococci (CPS) and 128 (49%) were coagulase-negative staphylococci (CNS). Eighteen different species of CNS were isolated, among which S. warneri, S. epidermidis and S. hyicus were the most frequent. The distribution of Staphylococcus species was different among herds: S. epidermidis was found in 8 herds, S. warneri was found in 7 herds, and S. hyicus in 6 herds. Some of the CNS species (S. saprophyticus ssp. saprophyticus, S. auricularis, S. capitis, and S. chromogenes) were isolated in only one of the farms. Genes related to production of enterotoxins were found in 66% (n=85) of all CNS and in 35% of the CPS isolates. For both CNS and CPS isolates, the most frequently identified enterotoxin genes were sea, seb, and sec; the prevalence of sea differed between CPS (9.5%) and CNS (35.1%) isolates. Staphylococcus warneri isolates showed a greater percentage of sea than seb, sec, or sed, whereas S. hyicus isolates showed a greater percentage of sea than sec. Over 60% of CNS belonged to 3 major species, which carried 62.2 to 81.3% of the enterotoxin genes. The high prevalence highlights the potential for food poisoning caused by these species. For

  5. A proteomics study of colostrum and milk from the two major small ruminant dairy breeds from the Canary Islands: a bovine milk comparison perspective.

    Science.gov (United States)

    Hernández-Castellano, Lorenzo E; Almeida, André M; Renaut, Jenny; Argüello, Anastasio; Castro, Noemí

    2016-08-01

    Colostrum and milk feeding are key factors for the newborn ruminant survival, affecting the future performance of the animal. Nowadays, there is an increasing interest in the potential of feeding newborn ruminants (mainly goat kids and lambs) with colostrum and milk from other more productive ruminant species (mainly cows). Although some studies regarding differences between colostrum and milk from these three species have been performed, herein we conduct for the first time a comparison using a proteomics 2-Dimensional Electrophoresis gel-based approach between these three ruminant species. In this study colostrum and milk samples from six Holstein cows, six Canarian sheep and six Majorera goats were used to determine the chemical composition, immunoglobulin G (IgG) and M (IgM) concentrations and proteomics profiles. Results showed that in general sheep colostrum and milk contained higher fat, protein and lactose percentages compared to bovine and goat samples. Additionally, no differences in the IgG or IgM concentrations were found among any of the three studied species, with the exception of sheep colostrum that showed the highest IgM concentration. With reference to the proteomics-based approach, some high abundant proteins such as serum albumin precursor, beta-caseins or different immunoglobulins components were found in colostrum, milk or even both. Nevertheless, differences in other proteins with immune function such as serotransferrin or lactoperoxidase were detected. This study shows that despite the similar immunoglobulin concentrations in colostrum and milk from the three studied species, differences in several immune components can be detected when these samples are studied using a proteomics approach. Finally, this study also provides a base for future investigation in colostrum and milk proteomics and metabolomics.

  6. LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples.

    Science.gov (United States)

    Dong, Xue; Zhou, Shiyue; Mechref, Yehia

    2016-06-01

    Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. The main component in milk oligosaccharides is free oligosaccharides. Since the proteins in milk are highly glycosylated, N-glycans in milk also play an import role. In this study, we investigated the permethylated free oligosaccharides and N-glycans extracted from bovine, goat, and human milks using LC-MS/MS. Quantitation profiles of free oligosaccharides and N-glycans were reported. The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. Human milk had more complex free oligosaccharides structures than the other two milk samples. Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) were associated with whey proteins extracted from bovine, goat, and human milk samples, respectively. Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. Permethylation of the glycan structures facilitated the interpretation of MS/MS. For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. This feature resulted in the identification of several isomers.

  7. Direct screening of tetracyclines in water and bovine milk using room temperature phosphorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Traviesa-Alvarez, J.M. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain); Costa-Fernandez, J.M. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain); Pereiro, R. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain); Sanz-Medel, A. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain)]. E-mail: asm@uniovi.es

    2007-04-18

    A fast and simple flow-through optosensor was designed and characterized for the direct screening of four tetracycline (TCC) antibiotics (tetracycline, oxytetracycline, chlortetracycline and doxycycline) in water and bovine milk samples. The proposed optosensor provides rapid binary yes/no overall responses, being appropriate for the screening of this family of antibiotics above or below a pre-set concentration threshold. The experimental set-up is based on a flow-injection manifold coupled on-line to a phosphorescence detector. Aliquots of the samples are pretreated with Eu(III) to form room temperature phosphorescent metal chelates and injected in the flow manifold. Those chelates are then on-line retained on a conventional flow-cell (packed with polymeric Amberlite XAD-4 particles) which is placed inside the cell holder of the phosphorimeter. After the emission is registered, the antibiotic-metal complexes are eluted from the packed resin with 1 M HCl (for milk samples a second regeneration step, using methanol, should be performed). A sample throughput of about 20 samples per hour was obtained. Optimum experimental conditions include a pH 9, a Eu(III) concentration of 2 x 10{sup -4} M and 8 mM sodium sulphite as chemical deoxygenant. The phosphorescence emitted by the europium-TCC complexes was measured at 394 and 617 nm for excitation and emission wavelengths, respectively. The unreliability region, given by the probability of false positives and false negatives, respectively (set at 5% in both cases) was in the range between 0.2 and 11.6 nM for detection of tetracyclines in water samples (at a cut-off level of 4 nM) and in the range between 165 and 238 nM for detection of tetracyclines in milk (cut-off level fixed at the normative EU level of 200 nM). Finally, the applicability of the proposed screening optosensor was tested for the reliable control of tetracyclines in contaminated and uncontaminated water and milk samples.

  8. Purification of bovine milk lactoperoxidase and investigation of antibacterial properties at different thiocyanate mediated.

    Science.gov (United States)

    Uğuz, M T; Ozdemir, H

    2005-01-01

    Bovine lactoperoxidase (LPO) was purified with amberlite CG 50 H+ resin, CM sephadex C-50 ion-exchange chromatography, and sephadex G-100 gel filtration chromatography from skim milk. The activity of lactoperoxidase was measured by using 2.2-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) diammonium salt (ABTS) as a choromogenic substrate at pH 6.0. Purification degree for the purified enzyme was controlled with SDS-PAGE and Rz value (A412/A280). Rz value for the purified LPO was 0.8. Km value at pH 6.0 at 20 degrees C for the LPO was 0.20 mM. Vmax value was 7.87 micromol/ml min at pH 6.0 at 20 degrees C. Bovine LPO showed high antibacterial activity in 100 mM thiocyanate--100 mM H2O2 medium for some pathogenic bacteria, such as Aeromonas hydrophila ATCC 7966, Micrococcus luteus LA 2971, Mycobacterium smegmatis RUT, Bacillus subtilis IMG 22, Pseudomonas pyocyanea, Bacillus subtilis var. niger ATCC 10, Pseudomonas aeruginosa ATCC 27853, Enterococcus faecalis ATCC 15753, Bacillus brevis FMC3, Klebsiella pneumoniae FMC 5, Corynebacterium xerosis UC 9165, Bacillus cereus EU, Bacillus megaterium NRS, Yersinia enterocolytica, Listeria monocytogenes scoot A, Bacillus megaterium EU, Bacillus megaterium DSM32, Klebsiella oxytocica, Staphylococcus aerogenes, Streptococcus faecalis, Mycobacterium smegmatis CCM 2067 and compared with well known antibacterial substances such as penicilline, ampicilline, amoxicillin-clavulanate and ceftriaxon. The LPO--100 mM thiocyanate--100 mM H2O2 system was purposed as an effective agent against many of the diseases causing organisms in human and animals.

  9. Prevalence and Antimicrobial Susceptibility Profiles of Pathogen Isolated from Bovine Mastitis Milk in Transylvania, Romania

    Directory of Open Access Journals (Sweden)

    Cosmina Bouari

    2016-11-01

    Full Text Available Mastitis in cows, one of the most common and economically important infectious diseases of dairy cattle, all over the world, with significant impact due to economic losses, occurs when the udder becomes inflamed because the leukocytes are released into the mammary gland usually in response to bacteria invasion of the teat canal. The main objective of this study was to evaluate the in vitro antimicrobial susceptibility of bacteria isolated from milk in order to design specific control programs for bovine mastitis in this area. A total of 204 milk samples aseptically collected both from farms and private owners were processed during May 2014 and March 2016 within the Microbiology Laboratory of the Faculty of Veterinary Cluj-Napoca, Romania. The microbiological examination was carried out by inoculation on blood agar and MacConkey medium. After the overnight incubation in aerobic conditions, the identification of the isolates was performed using microscopic, cultural and biochemical methods. Biochemical identification was based on API 20 Biomerieux system. Susceptibility to antibiotics was evaluated using Kirby Bauer disk diffusion method on Mueller-Hinton agar; the antibiotics were represented by Amoxicillin and Clavulanic Acid, Ceftiofur, Florfenicol, Mastidiscs, Enrofloxacin, Penicillin and Tetracycline. Staphylococcus spp. was the most common isolated pathogen, in 54.9% of the specimens, followed by Streptococcus spp. in 20.1%, Escherichia coli in 10.78%, Klebsiella spp. in 8.34%, Bacillus spp. in 5.88%. The most frequent associations were represented by staphylococci-streptococci in 62.7% of the samples, followed by streptococci-bacillus in 19.8% of the samples. The most important etiological agents identified were Staphylococcus aureus, S uberis, Streptococcus agalactiae, and Escherichia coli. Antimicrobial susceptibility test for the total isolates revealed good sensitivity to Enrofloxacin, Mastidiscs and Amoxicillin and Clavulanic Acid

  10. Sequence-based Association Analysis Reveals an MGST1 eQTL with Pleiotropic Effects on Bovine Milk Composition

    Science.gov (United States)

    Littlejohn, Mathew D.; Tiplady, Kathryn; Fink, Tania A.; Lehnert, Klaus; Lopdell, Thomas; Johnson, Thomas; Couldrey, Christine; Keehan, Mike; Sherlock, Richard G.; Harland, Chad; Scott, Andrew; Snell, Russell G.; Davis, Stephen R.; Spelman, Richard J.

    2016-01-01

    The mammary gland is a prolific lipogenic organ, synthesising copious amounts of triglycerides for secretion into milk. The fat content of milk varies widely both between and within species, and recent independent genome-wide association studies have highlighted a milk fat percentage quantitative trait locus (QTL) of large effect on bovine chromosome 5. Although both EPS8 and MGST1 have been proposed to underlie these signals, the causative status of these genes has not been functionally confirmed. To investigate this QTL in detail, we report genome sequence-based imputation and association mapping in a population of 64,244 taurine cattle. This analysis reveals a cluster of 17 non-coding variants spanning MGST1 that are highly associated with milk fat percentage, and a range of other milk composition traits. Further, we exploit a high-depth mammary RNA sequence dataset to conduct expression QTL (eQTL) mapping in 375 lactating cows, revealing a strong MGST1 eQTL underpinning these effects. These data demonstrate the utility of DNA and RNA sequence-based association mapping, and implicate MGST1, a gene with no obvious mechanistic relationship to milk composition regulation, as causally involved in these processes. PMID:27146958

  11. Simultaneous Determination of TetracyclinesResidues in Bovine Milk Samples by Solid Phase Extraction and HPLC-FL Method

    Directory of Open Access Journals (Sweden)

    Mehra Mesgari Abbasi

    2011-06-01

    Full Text Available Introduction:Tetracyclines (TCs are widely used in animal husbandry and their residues in milk may resultinharmful effects on human. The aim of this study was to investigate the presence of TCs residues in various bovine milk samples from local markets of Ardabil, Iran. Methods:One hundred and fourteen pasteurized, sterilized and raw milk samples were collected from markets of Ardabil. Tetracycline, Oxytetracycline and Chlortetracycline (TCs residues extraction carried out by Solid Phase Extraction method. Determination of TCs residues were performed by high performance liquid chromatography (HPLC method using Fluorescence detector.Results: The mean of total TCs residues in all samples (114 samples was 97.6 ±16.9ng/g and that of pasteurized, sterilized and raw milk samples were 87.1 ± 17.7, 112.0 ± 57.3 and 154.0 ± 66.3ng/g respectively. Twenty five point four percent of the all samples, and24.4%, 30% and 28.6% of the pasteurized, sterilized and raw milk samples, respectively had higher TCs residues than the recommended maximum levels (100ng/g. Conclusion:This study indicates the presence of tetracycline residues more than allowed amount. Regulatory authorities should ensure proper withdrawal period before milking the animals and definite supervisions are necessary on application of these drugs.

  12. Development of a loop-mediated isothermal amplification assay for the detection of Streptococcus agalactiae in bovine milk.

    Science.gov (United States)

    Bosward, Katrina L; House, John K; Deveridge, Amber; Mathews, Karen; Sheehy, Paul A

    2016-03-01

    Streptococcus agalactiae is a well-characterized bovine mastitis pathogen that is known to be highly contagious and capable of spreading rapidly in affected dairy herds. Loop-mediated isothermal amplification (LAMP) is a novel molecular diagnostic method that has the capability to provide rapid, cost-effective screening for pathogens to support on-farm disease control and eradication programs. In the current study, a LAMP test was developed to detect S. agalactiae in milk. The assay was validated on a bank of existing clinical mastitis milk samples that had previously been identified as S. agalactiae positive via traditional microbiological culture techniques and PCR. The LAMP assay was conducted on bacterial colonies and DNA extracted from milk in tube- and plate-based formats using multiple detection platforms. The 1-h assay conducted at 64 °C exhibited repeatability (coefficient of variation) of 2.07% (tube) and 8.3% (plate), sensitivity to ~20 pg of extracted DNA/reaction, and specificity against a panel of known bacterial mastitis pathogens. Of the 109 known S. agalactiae isolates assessed by LAMP directly from bacterial cells in culture, 108 were identified as positive, in accordance with PCR analysis. The LAMP analysis from the corresponding milk samples indicated that 104 of these milks exhibited a positive amplification curve. Although exhibiting some limitations, this assay provides an opportunity for rapid screening of milk samples to facilitate on-farm management of this pathogen.

  13. Short communication: Genetic correlation of bovine leukosis incidence with somatic cell score and milk yield in a US Holstein population.

    Science.gov (United States)

    Abdalla, E A; Weigel, K A; Byrem, T M; Rosa, G J M

    2016-03-01

    Bovine leukosis (BL) is a retroviral disease caused by the bovine leukosis virus (BLV), which affects only cattle. Dairy cows positive for BL produce less milk and have more days open than cows negative for BL. In addition, the virus also affects the immune system and causes weaker response to vaccines. Heritability estimates of BL incidence have been reported for Jersey and Holstein populations at about 0.08, indicating an important genetic component that can potentially be exploited to reduce the prevalence of the disease. However, before BL is used in selection programs, it is important to study its genetic associations with other economically important traits such that correlated responses to selection can be predicted. Hence, this study aimed to estimate the genetic correlations of BL with milk yield (MY) and with somatic cell score (SCS). Data of a commercial assay (ELISA) used to detect BLV antibodies in milk samples were obtained from Antel BioSystems (Lansing, MI). The data included continuous milk ELISA scores and binary milk ELISA results for 11,554 cows from 112 dairy herds across 16 US states. Continuous and binary milk ELISA were analyzed with linear and threshold models, respectively, together with MY and SCS using multitrait animal models. Genetic correlations (posterior means ± standard deviations) between BL incidence and MY were 0.17 ± 0.077 and 0.14 ± 0.076 using ELISA scores and results, respectively; with SCS, such estimates were 0.20 ± 0.081 and 0.17 ± 0.079, respectively. In summary, the results indicate that selection for higher MY may lead to increased BLV prevalence in dairy herds, but that the inclusion of BL (or SCS as an indicator trait) in selection indexes may help attenuate this problem.

  14. The effect of bovine milk lactoferrin on human breast cancer cell lines.

    Science.gov (United States)

    Duarte, D C; Nicolau, A; Teixeira, J A; Rodrigues, L R

    2011-01-01

    The evidence that biologically active food components are key environmental factors affecting the incidence of many chronic diseases is overwhelming. However, the full extent of such components in our diet is unknown, as is our understanding of their mechanisms of action. Beyond the interaction of these food components with the gut and intestinal immune functions, whey proteins such as lactoferrin are being tested as anticancer agents. Lactoferrin is an iron-binding protein that has been reported to inhibit several types of cancer. In the present work, the effects of bovine milk lactoferrin on human breast cancer HS578T and T47D cells were studied. The cells were either untreated or treated with lactoferrin concentrations ranging from 0.125 to 125 μM. Lactoferrin decreased the cell viability of HS578T and T47D by 47 and 54%, respectively, and increased apoptosis about 2-fold for both cell lines. Proliferation rates decreased by 40.3 and 63.9% for HS578T and T47D, respectively. For the T47D line, cell migration decreased in the presence of the protein. Although the mechanisms of action are not fully known, the results gathered in this work suggest that lactoferrin interferes with some of the most important steps involved in cancer development.

  15. Micellar nanotubes dispersed electrokinetic chromatography for the simultaneous determination of antibiotics in bovine milk.

    Science.gov (United States)

    Springer, Valeria H; Lista, Adriana G

    2012-07-01

    A method to determine four antibiotics for veterinary use (ciprofloxacin, enrofloxacin, florfenicol, and chloramphenicol) of different families (fluoroquinolones and amphenicols) in bovine milk was developed. The determination of the analytes was carried out using micellar electrokinetic capillary chromatography (MEKC) with a common sodium borate-SDS buffer solution containing single-walled carbon nanotubes (SWCNTs). In this way, a great improvement in the electrophoretic resolution and the separation efficiency was achieved compared to MEKC. An online reverse electrode polarity-stacking mode (REPSM) was carried out to enhance sensitivity. This step was performed in only 2 min and it allowed a stacked percentage of 103. That means that all the amount of injected analytes is effectively stacked. When this stacking procedure was combined with an off-line preconcentration step, based on SPE, analytes could be detected in lower concentration than the established maximum residue limits (MRLs). The LODs for the four compounds were between 6.8 and 13.8 μg L(-1) and the RSD values were between 1.1% and 6.6%. The whole method was applied to spiked real samples with acceptable precision and satisfactory recoveries.

  16. Detection of Mycobacterium tuberculosis and Mycobacterium avium Complexes by Real-Time PCR in Bovine Milk from Brazilian Dairy Farms.

    Science.gov (United States)

    Bezerra, André Vinícius Andrade; Dos Reis, Emily Marques; Rodrigues, Rogério Oliveira; Cenci, Alexander; Cerva, Cristine; Mayer, Fabiana Quoos

    2015-05-01

    Foodborne diseases are a public health problem worldwide. The consumption of contaminated raw milk has been recognized as a major cause of transmission of bovine tuberculosis to humans. Other mycobacteria that may be present in raw milk and may cause diseases are those belonging to the Mycobacterium avium complex. In this study, molecular biology tools were applied to investigate raw milk contamination with Mycobacterium spp. in family dairy farms from Rio Grande do Sul, southern Brazil. Furthermore, different variables related to the source of the milk, herd characteristics, and management were evaluated for their effect on milk contamination. Five hundred and two samples were analyzed, of which 354 were from the Northwest region (102 farms with samples from 93 bulk tanks and 261 animals) and 148 from the South region of the state (22 farms with samples from 23 bulk tanks and 125 animals). Among them, 10 (1.99%) and 7 (1.39%) were positive for Mycobacterium tuberculosis (9 confirmed as Mycobacterium bovis) and M. avium complexes, respectively. There was no difference in the frequencies of positive samples between the regions or the sample sources. Of the positive samples, 4 were collected from a bulk tank (1 positive for M. avium and 3 for M. tuberculosis). Moreover, 1 sample was positive concomitantly for M. tuberculosis and M. avium complexes. On risk analysis, no variable was associated with raw milk contamination by M. tuberculosis complex species. However, washing the udders of all animals and drying them with paper towels were weakly classified as risk factors for M. avium contamination. Positive samples were obtained from both animals and bulk tanks, which emphasizes the importance of tuberculosis control programs and provides evidence that milk monitoring can be used as a control practice. Moreover, the findings of this study reinforce the need for awareness of the problems of raw milk consumption among the general population.

  17. Effects of candidate gene polymorphisms on the detailed fatty acids profile determined by gas chromatography in bovine milk.

    Science.gov (United States)

    Pegolo, S; Cecchinato, A; Mele, M; Conte, G; Schiavon, S; Bittante, G

    2016-06-01

    Association analyses between candidate genes and bovine milk fatty acids can improve our understanding of genetic variation in milk fatty acid profiles and reveal potential opportunities to tailor milk fat composition through selection strategies. In this work, we investigated the association of 51 single nucleotide polymorphisms (SNP) selected from 37 candidate genes using a functional and positional approach, with 47 fatty acids, 9 fatty acid groups, and 5 Δ(9)-desaturation indices in milk samples from Brown Swiss cows. Individual milk samples were collected from 1,158 Italian Brown Swiss cows, and gas chromatography was used to obtain detailed milk fatty acid compositions. A GoldenGate assay system (Illumina, San Diego, CA) was used to perform genotype 96 selected SNP located in 54 genes across 22 chromosomes. In total, 51 polymorphic SNP in 37 candidate genes were retained for the association analysis. A Bayesian linear animal model was used to estimate the contribution of each SNP. A total of 129 tests indicated relevant additive effects between a given SNP and a single fatty acid trait; 38 SNP belonging to 30 genes were relevant for a total of 57 fatty acid traits. Most of the studied fatty acid traits (~81%) were relevantly associated with multiple SNP. Relevantly associated SNP were mainly found in genes related to fat metabolism, linked to or contained in previously identified quantitative trait loci for fat yield or content, or associated with genes previously identified in association analyses with milk fatty acid profiles in other cow breeds. The most representative candidate genes were LEP, PRL, STAT5A, CCL3, ACACA, GHR, ADRB2, LPIN1, STAT1, FABP4, and CSN2. In particular, relevant associations with SNP located on bovine chromosome 19 (BTA19) were found. Two candidate genes on BTA19 (CCL3 and ACACA) were relevantly associated with de novo short- and medium-chain fatty acids, likely explaining the high heritability values found for these fatty acids

  18. Iron and ferritin levels in the serum and milk of bovine leukemia virus-infected dairy cows

    Directory of Open Access Journals (Sweden)

    KOICHI eORINO

    2015-05-01

    Full Text Available Iron metabolism was examined in 15 bovine leukemia virus (BLV -infected dairy cows (2.6-7.8 years old. BLV infection was detected by measuring serum antibody titer against BLV virus antigen (gp51. The anti-BLV antibody titers of the BLV-infected cows were significantly higher in serum than in milk; a single serum-positive animal lacked detectable anti-BLV antibodies in its milk. Iron and ferritin concentrations also were significantly higher in serum than in milk. Although most of the BLV-infected dairy cows had past or present anamneses (such as inflammatory diseases, including intramammary infection, the milk ferritin concentrations of the infected cows were significantly lower than those of normal cows; serum ferritin concentrations did not differ significantly between these two groups. The anti-BLV antibody titers in milk samples showed significant correlation with serum iron concentrations. These results suggest that BLV infection affects iron homeostasis through iron metabolism in the dairy cow mammary gland.

  19. Effects of rapeseed and soybean oil dietary supplementation on bovine fat metabolism, fatty acid composition and cholesterol levels in milk.

    Science.gov (United States)

    Altenhofer, Christian; Spornraft, Melanie; Kienberger, Hermine; Rychlik, Michael; Herrmann, Julia; Meyer, Heinrich H D; Viturro, Enrique

    2014-02-01

    The main goal of this experiment was to study the effect of milk fat depression, induced by supplementing diet with plant oils, on the bovine fat metabolism, with special interest in cholesterol levels. For this purpose 39 cows were divided in three groups and fed different rations: a control group (C) without any oil supplementation and two groups with soybean oil (SO) or rapeseed oil (RO) added to the partial mixed ration (PMR). A decrease in milk fat percentage was observed in both oil feedings with a higher decrease of -1·14 % with SO than RO with -0·98 % compared with the physiological (-0·15 %) decline in the C group. There was no significant change in protein and lactose yield. The daily milk cholesterol yield was lower in both oil rations than in control ration, while the blood cholesterol level showed an opposite variation. The milk fatty acid pattern showed a highly significant decrease of over 10 % in the amount of saturated fatty acids (SFA) in both oil feedings and a highly significant increase in mono (MUFA) and poly (PUFA) unsaturated fatty acids, conjugated linoleic acids (CLA) included. The results of this experiment suggest that the feeding of oil supplements has a high impact on milk fat composition and its significance for human health, by decreasing fats with a potentially negative effect (SFA and cholesterol) while simultaneously increasing others with positive (MUFA, PUFA, CLA).

  20. Evaluation of milk yield losses associated with Salmonella antibodies in bulk tank milk in bovine dairy herds

    DEFF Research Database (Denmark)

    Nielsen, T D; Green, L E; Kudahl, Anne Margrethe Braad;

    2012-01-01

    The effect of Salmonella on milk production is not well established in cattle. The objective of this study was to investigate whether introduction of Salmonella into dairy cattle herds was associated with reduced milk yield and determine the duration of any such effect. Longitudinal data from 2005...... through 2009 were used, with data from 12 mo before until 18 mo after the estimated date of infection. Twenty-eight case herds were selected based on an increase in the level of Salmonella-specific antibodies in bulk-tank milk from ... was randomly allocated to the control herds. Hierarchical mixed effect models with the outcome test-day yield of energy-corrected milk (ECM)/cow were used to investigate daily milk yield before and after the estimated herd infection date for cows in parities 1, 2, and 3+. Control herds were used to evaluate...

  1. Rapid Profiling of Bovine and Human Milk Gangliosides by Matrix-Assisted Laser Desorption/Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry.

    Science.gov (United States)

    Lee, Hyeyoung; An, Hyun Joo; Lerno, Larry A; German, J Bruce; Lebrilla, Carlito B

    2011-08-15

    Gangliosides are anionic glycosphingolipids widely distributed in vertebrate tissues and fluids. Their structural and quantitative expression patterns depend on phylogeny and are distinct down to the species level. In milk, gangliosides are exclusively associated with the milk fat globule membrane. They may participate in diverse biological processes but more specifically to host-pathogen interactions. However, due to the molecular complexities, the analysis needs extensive sample preparation, chromatographic separation, and even chemical reaction, which makes the process very complex and time-consuming. Here, we describe a rapid profiling method for bovine and human milk gangliosides employing matrix-assisted desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). Prior to the analyses of biological samples, milk ganglioside standards GM3 and GD3 fractions were first analyzed in order to validate this method. High mass accuracy and high resolution obtained from MALDI FTICR MS allow for the confident assignment of chain length and degree of unsaturation of the ceramide. For the structural elucidation, tandem mass spectrometry (MS/MS), specifically as collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) were employed. Complex ganglioside mixtures from bovine and human milk were further analyzed with this method. The samples were prepared by two consecutive chloroform/methanol extraction and solid phase extraction. We observed a number of differences between bovine milk and human milk. The common gangliosides in bovine and human milk are NeuAc-NeuAc-Hex-Hex-Cer (GD3) and NeuAc-Hex-Hex-Cer (GM3); whereas, the ion intensities of ganglioside species are different between two milk samples. Kendrick mass defect plot yields grouping of ganglioside peaks according to their structural similarities. Gangliosides were further probed by tandem MS to confirm the compositional and structural assignments

  2. Comparison of culture and a multiplex probe PCR for identifying Mycoplasma species in bovine milk, semen and swab samples

    Science.gov (United States)

    Parker, Alysia M.; House, John K.; Hazelton, Mark S.; Bosward, Katrina L.; Sheehy, Paul A.

    2017-01-01

    Mycoplasma spp. are a major cause of mastitis, arthritis and pneumonia in cattle, and have been associated with reproductive disorders in cows. While culture is the traditional method of identification the use of PCR has become more common. Several investigators have developed PCR protocols to detect M. bovis in milk, yet few studies have evaluated other sample types or other important Mycoplasma species. Therefore the objective of this study was to develop a multiplex PCR assay to detect M. bovis, M. californicum and M. bovigenitalium, and evaluate its analytical performance against traditional culture of bovine milk, semen and swab samples. The PCR specificity was determined and the limit of detection evaluated in spiked milk, semen and swabs. The PCR was then compared to culture on 474 field samples from individual milk, bulk tank milk (BTM), semen and swab (vaginal, preputial, nose and eye) samples. Specificity analysis produced appropriate amplification for all M. bovis, M. californicum and M. bovigenitalium isolates. Amplification was not seen for any of the other Mollicutes or eubacterial isolates. The limit of detection of the PCR was best in milk, followed by semen and swabs. When all three Mycoplasma species were present in a sample, the limit of detection increased. When comparing culture and PCR, overall there was no significant difference in the proportion of culture and PCR positive samples. Culture could detect significantly more positive swab samples. No significant differences were identified for semen, individual milk or BTM samples. PCR identified five samples with two species present. Culture followed by 16S-23S rRNA sequencing did not enable identification of more than one species. Therefore, the superior method for identification of M. bovis, M. californicum and M. bovigenitalium may be dependent on the sample type being analysed, and whether the identification of multiple target species is required. PMID:28264012

  3. Proteomics data in support of the quantification of the changes of bovine milk proteins during mammary gland involution

    Directory of Open Access Journals (Sweden)

    Irina Boggs

    2016-09-01

    Full Text Available Here we provide data from three proteomics techniques; two-dimensional electrophoresis (2-DE followed by identification of selected spots using PSD MALDI-TOF MS/MS, one-dimensional gel electrophoresis followed by LC-MS/MS analysis of gel slices (GeLC and dimethyl isotopic labelling of tryptic peptides followed by Orbitrap MS/MS (DML, to quantify the changes in the repertoire of bovine milk proteins that occurs after drying off. We analysed skim milk and whey sampled at day 0 and either day 3 or day 8 after drying off. These analyses identified 45 spots by MALDI-TOF, 51 proteins by GeLC and 161 proteins by DML, for which the detailed data work-up is presented as three Excel files. The data supplied in this article supports the accompanying publication “Changes in the repertoire of bovine milk proteins during mammary involution” (Boggs et al., 2015 [1]. Data are available via ProteomeXchange with identifiers ProteomeXchange: PXD003110 and ProteomeXchange: PXD003011.

  4. Effects of bovine mammary gland biopsy and increased milking frequency on post-procedure udder health, histology, and milk yield.

    Science.gov (United States)

    Lima, J A M; Ruas, J R M; Vasconcelos, A C; Silper, B F; Lana, A M Q; Gheller, V A; Saturnino, H M; Reis, R B; Coelho, S G

    2016-05-01

    Sixteen cows in early lactation were randomly distributed into two groups in order to evaluate the effects of mammary biopsies and increased milking frequency on tissue characteristics, post-biopsy udder health and histology. One group was milked twice a day (2×) starting on the 2nd day after calving, until 28 days in milk (DIM). The other group was milked four times a day (4×) from two to 21 DIM, and twice a day (2×) from 22 to 28 DIM. On days 2, 7, 14, 21, and 28 postpartum, one fragment of secretory tissue was collected from one mammary quarter at a time. Collections were alternated between the four mammary quarters per collection day. A total of 80 mammary tissue samples were collected. Qualitative and quantitative analyses of the tissues were conducted by histologic examination. Animal health was assessed by observation of feed intake behavior immediately after biopsy, and weight and body condition score before and one week after biopsy. Udder health was assessed daily from calving to 60 DIM with California Mastitis Test (CMT) and by noting alterations in the milk such as blood, milk clots, blood clots, clinical signs of mastitis. Milk composition and somatic cell count (SCC) were analyzed before and after the biopsies. Milk production was evaluated before biopsy, on the day of biopsy, and after the biopsy. An average of 10 fields at 40× magnification was obtained from each sample. There were no evident changes in mammary morphology as a result of milking two or four times/day at any of the evaluated time points. Biopsy wounds healed rapidly without infection. Intramammary bleeding and CMT alterations were observed in 96% and 75% of the biopsied mammary quarters, respectively. Clinical mastitis was diagnosed in 12% of the biopsied quarters. Different milking frequencies had no effect on the frequency and duration of post-biopsy alterations. Milk production decreased after biopsies done on days 2 for 2× and 4× groups, but it returned to pre-biopsy values

  5. Bovine milk proteome: Quantitative changes in normal milk exosomes, milk fat globule membranes and whey proteomes resulting from Staphylococcus aureus mastitis

    Science.gov (United States)

    Knowledge of milk protein composition/expression in healthy cows and cows with mastitis will provide information important for the dairy food industry, mammary biology and immune function in the mammary gland. To facilitate maximum protein discovery, milk was fractioned into whey, milk fat globule ...

  6. Magnetic bead and gold nanoparticle probes based immunoassay for β-casein detection in bovine milk samples.

    Science.gov (United States)

    Li, Y S; Meng, X Y; Zhou, Y; Zhang, Y Y; Meng, X M; Yang, L; Hu, P; Lu, S Y; Ren, H L; Liu, Z S; Wang, X R

    2015-04-15

    In this work, a double-probe based immunoassay was developed for rapid and sensitive determination of β-casein in bovine milk samples. In the method, magnetic beads (MBs), employed as supports for the immobilization of anti-β-casein polyclonal antibody (PAb), were used as the capture probe. Colloidal gold nanoparticles (AuNPs), employed as a bridge for loading anti-β-casein monoclonal antibody (McAb) and horseradish peroxidase (HRP), were used as the amplification probe. The presence of β-casein causes the sandwich structures of MBs-PAb-β-casein-McAb-AuNPs through the interaction between β-casein and the anti-β-casein antibodies. The HRP, used as an enzymatic-amplified tracer, can catalytically oxidize the substrate 3,3',5,5'-tetramethylbenzidine (TMB), generating optical signals that are proportional to the quantity of β-casein. The linear range of the immunoassay was from 6.5 to 1520ngmL(-1). The limit of detection (LOD) was 4.8ngmL(-1) which was 700 times lower than that of MBs-antibody-HRP based immunoassay and 6-7 times lower than that from the microplate-antibody-HRP based assay. The recoveries of β-casein from bovine milk samples were from 95.0% to 104.3% that had a good correlation coefficient (R(2)=0.9956) with those obtained by an official standard Kjeldahl method. For higher sensitivity, simple sample pretreatment and shorter time requirement of the antigen-antibody reaction, the developed immunoassay demonstrated the viability for detection of β-casein in bovine milk samples.

  7. Survival rate of campylobacter coli strains in sterile buffalo and bovine milk Tasas de sobrevida de Campylobacter coli en leche de búfalo y de bovino

    Directory of Open Access Journals (Sweden)

    A. TRESIERRA-AYALA

    2001-01-01

    Full Text Available The survival rate of five Campylobacter coli strains, isolated from bovine faeces (3 and from buffalo faeces (2, was studied in sterile buffalo and bovine milk kept at 4°C under aerobic conditions. All strains lost their viability substantially more rapidly in buffalo milk than in bovine milkSe determinó, en leche de bovino y de búfalo autoclavadas y mantenidas a 4ºC bajo condiciones de aerobiosis, la sobrevida de cinco cepas de Campylobacter coli aisladas de fecas de bovino (3 y de búfalo (2. Todas las cepas perdieron su viabilidad más rápidamente en leche de búfalo que en leche bovina

  8. Evidence of penetration of the bovine teat duct by Escherichia coli in the interval between milkings.

    Science.gov (United States)

    Bramley, A J; Godinho, K S; Grindal, R J

    1981-10-01

    In 3 consecutive experiments, each using 20 cows, the application of Escherichia coli to teat ends after milking led to high rates of intramammary infection. These infections were not prevented by disinfection of the teats before milking, by the installation of shields in the short milk tubes of th milking cluster or by the use of an individual quarter milking cluster. Rates of infection were significantly lower when teat contamination was applied 1 h before milking compared to contamination applied immediately after milking. These data suggest that penetration of te teat duct by the E. coli occurred in the period between contamination and milking. Seventy four percent of infections occurred in hindquarters and there were variations in the susceptibility of cows to infection.

  9. Chemical changes in bovine milk fat globule membrane caused by heat treatment and homogenization of whole milk.

    Science.gov (United States)

    Lee, Sung Je; Sherbon, John W

    2002-11-01

    The effects of heat treatment and homogenization of whole milk on chemical changes in the milk fat globule membrane (MFGM) were investigated. Heating at 80 degrees C for 3-18 min caused an incorporation of whey proteins, especially beta-lactoglobulin (beta-Ig), into MFGM, thus increasing the protein content of the membrane and decreasing the lipid. SDS-PAGE showed that membrane glycoproteins, such as PAS-6 and PAS-7, had disappeared or were weakly stained in the gel due to heating of the milk. Heating also decreased free sulphydryl (SH) groups in the MFGM and increased disulphide (SS) groups, suggesting that incorporation of beta-Ig might be due to association with membrane proteins via disulphide bonds. In contrast, homogenization caused an adsorption of caseins to the MFGM but no binding of whey proteins to the MFGM without heating. Binding of caseins and whey proteins and loss of membrane proteins were not significantly different between milk samples that were homogenized before and after heating. Viscosity of whole milk was increased when milk was treated with both homogenization and heating.

  10. In vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on Prototheca zopfii strains isolated from bovine milk.

    Science.gov (United States)

    Salerno, Tatiana; Ribeiro, Márcio Garcia; Langoni, Hélio; Siqueira, Amanda Keller; Costa, Elizabeth Oliveira da; Melville, Priscilla Anne; Bueno, Válter Ferreira Félix; Yamamura, Aline Artioli Machado; Roesler, Uwe; da Silva, Aristeu Vieira

    2010-04-01

    Prototheca zopfii has been considered one of the most important causes of environmental mastitis in Brazil. These algae are refractory to conventional therapy and cause great damage to the mammary gland. The present study evaluated the in vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on 27 P. zopfii strains isolated from the milk of cattle. Low concentrations of sodium hypochlorite (0.0390625-0.15625%) and iodine (0.15625-0.625%) were effective against the isolates. These antiseptics may be recommended for hygiene routines, pre and postdipping and cauterization of bovine mammary glands infected by P. zopfii.

  11. Effect of dairy farming system, herd, season, parity, and days in milk on modeling of the coagulation, curd firming, and syneresis of bovine milk.

    Science.gov (United States)

    Bittante, G; Cipolat-Gotet, C; Malchiodi, F; Sturaro, E; Tagliapietra, F; Schiavon, S; Cecchinato, A

    2015-04-01

    The objectives of this study were to characterize the variation in curd firmness model parameters obtained from coagulating bovine milk samples, and to investigate the effects of the dairy system, season, individual farm, and factors related to individual cows (days in milk and parity). Individual milk samples (n = 1,264) were collected during the evening milking of 85 farms representing different environments and farming systems in the northeastern Italian Alps. The dairy herds were classified into 4 farming system categories: traditional system with tied animals (29 herds), modern dairy systems with traditional feeding based on hay and compound feed (30 herds), modern dairy system with total mixed ration (TMR) that included silage as a large proportion of the diet (9 herds), and modern dairy system with silage-free TMR (17 herds). Milk samples were analyzed for milk composition and coagulation properties, and parameters were modeled using curd firmness measures (CFt) collected every 15 s from a lacto-dynamographic analysis of 90 min. When compared with traditional milk coagulation properties (MCP), the curd firming measures showed greater variability and yielded a more accurate description of the milk coagulation process: the model converged for 93.1% of the milk samples, allowing estimation of 4 CFt parameters and 2 derived traits [maximum CF (CF(max)) and time from rennet addition to CF(max) (t(max))] for each sample. The milk samples whose CFt equations did not converge showed longer rennet coagulation times obtained from the model (RCT(eq)) and higher somatic cell score, and came from less-productive cows. Among the sources of variation tested for the CFt parameters, dairy herd system yielded the greatest differences for the contrast between the traditional farm and the 3 modern farms, with the latter showing earlier coagulation and greater instant syneresis rate constant (k(SR)). The use of TMR yielded a greater tmax because of a higher instant curd

  12. Whole genome scan to detect quantitative trait loci for bovine milk protein composition

    NARCIS (Netherlands)

    Schopen, G.C.B.; Koks, P.D.; Arendonk, van J.A.M.; Bovenhuis, H.; Visker, M.H.P.W.

    2009-01-01

    The objective of this study was to perform a whole genome scan to detect quantitative trait loci (QTL) for milk protein composition in 849 Holstein–Friesian cows originating from seven sires. One morning milk sample was analysed for the major milk proteins using capillary zone electrophoresis. A gen

  13. Molecular screening of bovine raw milk for the presence of Shiga toxin-producing Escherichia coli (STEC on dairy farms

    Directory of Open Access Journals (Sweden)

    Tatiane Vendramin

    2014-09-01

    Full Text Available Milkborne transmission of Shiga toxin- producing Escherichia coli (STEC has raised considerable concern due to recent outbreaks worldwide and poses a threat to public health. The aim of this study was to develop a sensitive and specific multiplex PCR assay to detect the presence of STEC in bovine raw milk. To identify E. coli (ATCC 25922 contamination, the gene uspA was used, and PCR sensitivity and specificity were accessed by testing diluted samples ranging from 2 to 2.0 × 10(6 CFU/mL. To detect STEC, the stx1 and stx2 genes were selected as targets. After reaction standardization, the multiplex assay was tested in raw milk collected from 101 cows on dairy farms. PCR assay for E. coli detection had a specificity of 100% and sensitivity of 79% (P<0.0001, with a lower detection limit of 2 CFU/mL. Multiplex PCR assay had 100% sensitivity for E. coli positive raw milk samples, and 31.1% were contaminated with STEC, 28.3% of stx2, and 1.9% of stx1. The multiplex PCR assay described in the present study can be employed to identify and screen E. coli harboring stx1 and stx2 genes in raw milk on dairy farms and in industries.

  14. Determination of creatinine, uric and ascorbic acid in bovine milk and orange juice by hydrophilic interaction HPLC.

    Science.gov (United States)

    Zuo, Ruiting; Zhou, Si; Zuo, Yuegang; Deng, Yiwei

    2015-09-01

    Creatinine (Cr), uric (UA) and ascorbic acid (AA) are common constituents in human fluids. Their abnormal concentrations in human fluids are associated with various diseases. Thus, apart from the endogenous formation in human body, it is also important to examine their sources from food products. In this study, a rapid and accurate HILIC method was developed for simultaneous determination of Cr, UA and AA in bovine milk and orange juice. Milk samples were pretreated by protein precipitation, centrifugation and filtration, followed by HPLC separation and quantification using a Waters Spherisorb S5NH2 column. The developed method has been successfully applied to determine the concentration of UA, AA and Cr in milk and fruit juice samples. The milk samples tested were found to contain UA and creatinine in the concentration range of 24.1-86.0 and 5.07-11.2 μg mL(-1), respectively. The orange juices contain AA over 212 μg mL(-1).

  15. A multiresidue liquid chromatographic/tandem mass spectrometric method for the detection and quantitation of 15 nonsteroidal anti-inflammatory drugs (NSAIDs) in bovine meat and milk.

    Science.gov (United States)

    van Pamel, Els; Daeseleire, Els

    2015-06-01

    This study concerns a validated liquid chromatographic/tandem mass spectrometric (LC-MS/MS) multiresidue method for the simultaneous detection, identification, and quantitation of 15 nonsteroidal anti-inflammatory drugs (NSAIDs) in bovine meat and milk. The NSAIDs considered are carprofen, diclofenac, flufenamic acid, flunixin (5-hydroxyflunixin as marker metabolite in milk), ketoprofen, mefenamic acid, meloxicam, 4-methylaminoantipyrine (marker metabolite of metamizole in meat and milk), naproxen, niflumic acid, phenylbutazone (and metabolite oxyphenbutazone), ramifenazone, salicylic acid, and tolfenamic acid. These compounds were chosen as representatives of different chemical subclasses of NSAIDs. Flunixin-d3, diclofenac-d4, 4-aminoantipyrine-d3, and phenylbutazone-d10 were used as internal standards. Performance characteristics were validated according to the Commission Decision 2002/657/EC (Off J Eur Communities, L221: 8-36). Recovery percentages varied between 81 and 114% for bovine meat and between 79 and 118% for milk. Repeatability percentages were within the range of 1-12% for meat and between 1 and 17% for milk, whereas the intralaboratory reproducibility varied between 3 and 19% for meat and between 3 and 23% for milk. The decision limit and the detection capability for bovine meat were within the range of 0.5-579 μg kg(-1)and 0.6-642 μg kg(-1), respectively. Those for milk were within the range of 0.12-55 μg kg(-1) and 0.14-61 μg kg(-1), respectively. The methods developed were successfully applied for proficiency test samples and routine samples analyzed in the laboratory. The methodology concerns fast, user-friendly, and sensitive methods, which can be easily extended for other compounds and matrices. In general, such multiresidue methods contribute to the reduction of human exposure to these veterinary drug residues by consumption of contaminated bovine-derived products such as meat and milk.

  16. Quantification of individual fatty acids in bovine milk by infrared spectroscopy and chemometrics: understanding predictions of highly collinear reference variables.

    Science.gov (United States)

    Eskildsen, C E; Rasmussen, M A; Engelsen, S B; Larsen, L B; Poulsen, N A; Skov, T

    2014-12-01

    Predicting individual fatty acids (FA) in bovine milk from Fourier transform infrared (FT-IR) measurements is desirable. However, such predictions may rely on covariance structures among individual FA and total fat content. These covariance structures may change with factors such as breed and feed, among others. The aim of this study was to estimate how spectral variation associated with total fat content and breed contributes to predictions of individual FA. This study comprised 890 bovine milk samples from 2 breeds (455 Holstein and 435 Jersey). Holstein samples were collected from 20 Danish dairy herds from October to December 2009; Jersey samples were collected from 22 Danish dairy herds from February to April 2010. All samples were from conventional herds and taken while cows were housed. Moreover, in a spiking experiment, FA (C14:0, C16:0, and C18:1 cis-9) were added (spiked) to a background of commercial skim milk to determine whether signals specific to those individual FA could be obtained from the FT-IR measurements. This study demonstrated that variation associated with total fat content and breed was responsible for successful FT-IR-based predictions of FA in the raw milk samples. This was confirmed in the spiking experiment, which showed that signals specific to individual FA could not be identified in FT-IR measurements when several FA were present in the same mixture. Hence, predicted concentrations of individual FA in milk rely on covariance structures with total fat content rather than absorption bands directly associated with individual FA. If covariance structures between FA and total fat used to calibrate partial least squares (PLS) models are not conserved in future samples, these samples will show incorrect and biased FA predictions. This was demonstrated by using samples of one breed to calibrate and samples of the other breed to validate PLS models for individual FA. The 2 breeds had different covariance structures between individual FA and

  17. Association between antibody status to bovine herpesvirus 1 and quality of milk in dairy herds in Poland.

    Science.gov (United States)

    Rola, J G; Larska, M; Grzeszuk, M; Rola, J

    2015-02-01

    Bovine herpesvirus 1 (BoHV1) is one of the most important pathogens of cattle; however, its effect on somatic cell count and milk components is not completely understood. The aim of the current study was to examine the effect of BoHV1 infection on quality of bovine bulk tank milk (BTM). A total of 1,790 individual blood samples collected at 28 dairy farms were used to determine the BoHV1 infection status of the herds with ELISA tests. The quality parameters of milk were evaluated by instrumental methods with BTM samples collected at monthly intervals from May 2011 to May 2012. The statistical analysis was performed to study the associations between BoHV1 herd status, quality of BTM, and herd-specific parameters. The risk factors influencing bulk milk somatic cell count (BMSCC) were estimated using the multivariable mixed-effects maximum likelihood regression model. The true prevalences of BoHV1 infection at the animal and herd levels were 49.3 and 64.6%, respectively. The average BMSCC differed significantly between the herds grouped accordingly to their BoHV1 infection status. Interestingly, the highest BMSCC was observed in the vaccinated herds (240.3×10(3) cells/mL). Additionally, the BoHV1 herd status had a significant effect on the fat content of BTM. The largest herds that were investigated had a BoHV1 seroprevalence over 30%. The herd status was considerably influenced by the numbers of cows in the herds. Besides, no significant differences in total bacterial count or protein content in milk from BoHV1-infected und uninfected herds were observed. An increase in BMSCC was observed during summer compared with the winter months regardless of the BoHV1 status of the herds. In the final multivariable regression model, the main risk factors associated with BMSCC were BoHV1 herd status, the percentage of BoHV1 infected animals in a herd, the number of cows in a herd, and the season. Our study suggests that BoHV1 infection may influence BMSCC levels, which are key

  18. Identification of Candida species isolated from bovine mastitic milk and their in vitro hemolytic activity in Western Turkey.

    Science.gov (United States)

    Seker, Esra

    2010-04-01

    In this study, identification of 207 Candida isolates, previously isolated from mastitic bovine quarter milk samples at the level of genus, was made using API 20 C AUX system. The most frequently isolated species were Candida krusei (34.8%), followed by Candida rugosa (16.4%), Candida kefyr (12.6%), Candida albicans (10.1%), and Candida tropicalis (9.2%). Less common isolates were Candida zeylanoides (5.8%), Candida parapsilosis (4.3%), Candida guilliermondii (3.4%), Candida famata (1.9%), and Candida glabrata (1.5%). Additionally, in vitro hemolytic activity of all Candida strains were also examined in the present study. C. krusei (72 isolates), C. kefyr (26), C. albicans (21), C. tropicalis (19), C. zeylanoides (12), and C. glabrata (3) demonstrated both alpha and beta hemolysis at 48-h postinoculation. Only alpha hemolysis was detected in C. rugosa (34), C. guilliermondii (7), and C. famata (4), while C. parapsilosis (9) did not show any hemolytic activity after incubation for 72 h. Statistically significant difference (P hemolytic activities of Candida strains. The hemolytic activities of C. zeylanoides, C. albicans and C. kefyr were higher than other strains. This is the first study to describe variable hemolysis types exhibited by different Candida strains isolated from bovine mastitic milk in Turkey.

  19. Predicting bovine milk fat composition using infrared spectroscopy based on milk samples collected in winter and summer

    NARCIS (Netherlands)

    Rutten, M.J.M.; Bovenhuis, H.; Hettinga, K.A.; Valenberg, van H.J.F.; Arendonk, van J.A.M.

    2009-01-01

    It has recently been shown that Fourier transform infrared spectroscopy has potential for the prediction of detailed milk fat composition, even based on a limited number of observations. Therefore, there seems to be an opportunity for improvement by means of using more observations. The objective of

  20. Milk losses due to bovine tropical theileriosis (Theileria annulata infection) in Algeria

    OpenAIRE

    Ouarda Ayadi; Mohamed Gharbi; Mohamed Cherif Benchikh Elfegoun

    2016-01-01

    The authors studied the impact of tropical theileriosis onset on milk yield decrease in 10 local bred cows in Skikda (Northern Algeria) during 2015 summer season. The milk yield decrease estimated weekly during two months was 2.76 L/day/cow corresponding to 31.92% of the total milk yield. This decrease corresponds to 110.5 Algerian Dinars (1.02 US$)/day/diseased cow. The relative variation of milk yield showed a dramatic decrease from 82.72% to 0.76% at Day 21 then became constant. Further st...

  1. Some nutritional effects of folate-binding protein in bovine milk on the bioavailability of folate to rats

    Energy Technology Data Exchange (ETDEWEB)

    Tani, M.; Iwai, K.

    1984-04-01

    The excretions of folate compounds into both the urine and bile were investigated in rats after the administration of pteroylglutamic acid (PteGlu) with or without the folate-binding protein (FBP) prepared from bovine milk. When the sample solution, containing either free or bound (/sup 3/H)PteGlu (i.e., bound to the FBP from milk), was delivered to rats intragastrically via oral intubation, the amounts of (/sup 3/H)PteGlu excreted into the feces did not change. On the other hand, the urinary excretion of /sup 3/H-labeled folate compounds, especially (/sup 3/H)5-methyltetrahydrofolic acid (5-CH/sub 3/-H/sub 4/PteGlu), after the administration of bound (/sup 3/H)PteGlu was significantly lower (P less than 0.01) than that after the administration of free (/sup 3/H)PteGlu. The urinary excretion of (/sup 3/H)5-CH/sub 3/-H/sub 4/PteGlu was directly proportional to the initial amount of free (/sup 3/H)PteGlu administered. The similar effect of FBP was also observed when the biliary excretion of /sup 3/H-labeled folate compounds was investigated in situ. Furthermore, the incorporation of (/sup 3/H)PteGlu into folate-requiring intestinal microorganisms was considerably reduced when it was bound to FBP. These results suggest that milk FBP has some nutritional effects on the bioavailability of folate in vivo.

  2. A simple, rapid and reliable enzyme-linked immunosorbent assay for the detection of bovine virus diarrhoea virus (BVDV) specific antibodies in cattle serum, plasma and bulk milk

    NARCIS (Netherlands)

    Kramps, J.A.; Maanen, van C.; Wetering, van de G.; Stienstra, G.; Quak, S.; Brinkhof, J.; Ronsholt, L.; Nylin, B.

    1997-01-01

    To detect Bovine Virus Diarrhoea Virus (BVDV)-specific antibodies in cattle serum, plasma and bulk milk, a simple, reliable and rapid blocking ELISA ("Ceditest") has been developed using two monoclonal antibodies ("WB112" and "WB103") directed to different highly conserved epitopes on the non-struct

  3. Comparative proteomic exploration of whey proteins in human and bovine colostrum and mature milk using iTRAQ-coupled LC-MS/MS.

    Science.gov (United States)

    Yang, Mei; Cao, Xueyan; Wu, Rina; Liu, Biao; Ye, Wenhui; Yue, Xiqing; Wu, Junrui

    2017-02-20

    Whey, an essential source of dietary nutrients, is widely used in dairy foods for infants. A total of 584 whey proteins in human and bovine colostrum and mature milk were identified and quantified by the isobaric tag for relative and absolute quantification (iTRAQ) proteomic method. The 424 differentially expressed whey proteins were identified and analyzed according to gene ontology (GO) annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway, and multivariate statistical analysis. Biological processes principally involved biological regulation and response to stimulus. Major cellular components were extracellular region part and extracellular space. The most prevalent molecular function was protein binding. Twenty immune-related proteins and 13 proteins related to enzyme regulatory activity were differentially expressed in human and bovine milk. Differentially expressed whey proteins participated in many KEGG pathways, including major complement and coagulation cascades and in phagosomes. Whey proteins show obvious differences in expression in human and bovine colostrum and mature milk, with consequences for biological function. The results here increase our understanding of different whey proteomes, which could provide useful information for the development and manufacture of dairy products and nutrient food for infants. The advanced iTRAQ proteomic approach was used to analyze differentially expressed whey proteins in human and bovine colostrum and mature milk.

  4. Diagnostic performance of an indirect enzyme-linked immunosorbent assay (ELISA) to detect bovine leukemia virus antibodies in bulk-tank milk samples.

    Science.gov (United States)

    Nekouei, Omid; Durocher, Jean; Keefe, Greg

    2016-07-01

    This study assessed the diagnostic performance of a commercial ELISA for detecting bovine leukemia virus antibodies in bulk-tank milk samples from eastern Canada. Sensitivity and specificity of the test were estimated at 97.2% and 100%, respectively. The test was recommended as a cost-efficient tool for large-scale screening programs.

  5. Kinetics of local and systemic isoforms of serum amyloid A in bovine mastitic milk

    NARCIS (Netherlands)

    Jacobsen, S.; Niewold, T.A.; Kornalijnslijper, E.; Toussaint, M.J.M.; Gruys, E.

    2005-01-01

    The aim of the present study was to characterise the serum amyloid A (SAA) response to intramammary inoculation of Escherichia coli and to examine the distribution of hepatically and extrahepatically produced SAA isoforms in plasma and milk from cows with mastitis. Milk and plasma SAA concentrations

  6. MicroRNA expression profiles of bovine milk exosomes in response to Staphylococcus aureus infection

    Science.gov (United States)

    Background: Milk exosomes are a rich source of microRNAs (miRNAs) that are protected from degradation. Ingestion of milk and subsequent absorption of miRNAs into recipient cells by endocytosis may play a role in the regulation of neonatal innate and adaptive immunity. In contrast, the miRNA content ...

  7. Effect of total proteose-peptone content on the variability of bovine milk foaming property

    Directory of Open Access Journals (Sweden)

    Arianna Buccioni

    2013-02-01

    Full Text Available Several authors demonstrated a strong linkage between proteose-peptones content and foaming properties of cow milk; this is of great interest for Italian dairy industries to create a new line of fresh milk characterized by a particular foaming property and, hence, particularly appreciate in catering industry. The aim of this trial was to quantify the relation between total concentration of proteose-peptones and the entity of foaming attitude in cow fresh milk. Ninety samples of raw bulk milk were analysed for proteose-peptones content, plasmin activity, fatty acid profile and foaming attitude. A negative relation was found among proteose-peptones percentage and foaming attitude which decreased with the increase of plasmin activity and somatic cell content in milk.

  8. Oral Administration of Bovine Milk from Cows Hyperimmunized with Intestinal Bacterin Stimulates Lamina Propria T Lymphocytes to Produce Th1-Biased Cytokines in Mice

    Directory of Open Access Journals (Sweden)

    Yuanyuan Wang

    2014-03-01

    Full Text Available The goal of this study was to examine the effects of oral administration of bovine milk from cows hyperimmunized with a proprietary bacterin (immune milk “Sustaina” on mucosal immunity in the intestine of adult mice. C57BL/6 mice were orally given immune or control milk for two weeks, and then lymphocyte population and the cytokine production in lamina propria of colon in normal mice and mice induced colitis by dextran sulphate sodium (DSS were detected. We found that the levels of IFN-γ and IL-10 increased, but the levels of IL-17A and IL-4, decreased in lamina propria of colon in immune milk-fed mice as compared with those in control milk-fed mice. Interestingly, oral administration of immune milk partially improved the acute colitis induced by DSS. The levels of TNF-α and IFN-γ increased, but IL-6, IL-17A and IL-4 decreased in lamina propria (LP of colon in immune milk-fed mice with DSS-induced colitis. Our results suggest that immune milk may stimulate CD4+ T cells to polarize towards a Th1 type response, but contrarily suppress Th17 and Th2 cells responses in large intestinal LP of mice. The results indicate that this kind of immune milk has is able to promote the maintainance of intestinal homeostasis and enhance protection against infection, and could alleviate the symptoms of acute colitis in mice.

  9. Effectiveness of mid-infrared spectroscopy to predict the color of bovine milk and the relationship between milk color and traditional milk quality traits.

    Science.gov (United States)

    McDermott, A; Visentin, G; McParland, S; Berry, D P; Fenelon, M A; De Marchi, M

    2016-05-01

    The color of milk affects the subsequent color features of the resulting dairy products; milk color is also related to milk fat concentration. The objective of the present study was to quantify the ability of mid-infrared spectroscopy (MIRS) to predict color-related traits in milk samples and to estimate the correlations between these color-related characteristics and traditional milk quality traits. Mid-infrared spectral data were available on 601 milk samples from 529 cows, all of which had corresponding gold standard milk color measures determined using a Chroma Meter (Konica Minolta Sensing Europe, Nieuwegein, the Netherlands); milk color was expressed using the CIELAB uniform color space. Separate prediction equations were developed for each of the 3 color parameters (L*=lightness, a*=greenness, b*=yellowness) using partial least squares regression. Accuracy of prediction was determined using both cross validation on a calibration data set (n=422 to 457 samples) and external validation on a data set of 144 to 152 samples. Moderate accuracy of prediction was achieved for the b* index (coefficient of correlation for external validation=0.72), although poor predictive ability was obtained for both a* and L* indices (coefficient of correlation for external validation of 0.30 and 0.55, respectively). The linear regression coefficient of the gold standard values on the respective MIRS-predicted values of a*, L*, and b* was 0.81, 0.88, and 0.96, respectively; only the regression coefficient on L* was different from 1. The mean bias of prediction (i.e., the average difference between the MIRS-predicted values and gold standard values in external validation) was not different from zero for any of 3 parameters evaluated. A moderate correlation (0.56) existed between the MIRS-predicted L* and b* indices, both of which were weakly correlated with the a* index. Milk fat, protein, and casein were moderately correlated with both the gold standard and MIRS-predicted values

  10. Development of a single nucleotide polymorphism genotyping microarray platform for the identification of bovine milk protein genetic polymorphisms.

    Science.gov (United States)

    Chessa, S; Chiatti, F; Ceriotti, G; Caroli, A; Consolandi, C; Pagnacco, G; Castiglioni, B

    2007-01-01

    The objective of this study was to develop and validate a fast method for typing the main mutations of bovine milk protein genes by using microarray technology. An approach based on the ligation detection reaction (LDR) and a universal array (UA) was used. Polymorphisms in both the coding and noncoding sequences of alpha(S1)-casein, beta-casein, kappa-casein, and beta-lactoglobulin genes were considered because of their well-known effects on milk composition and cheese production. A total of 22 polymorphic sites, corresponding to 21 different variants, were included in the diagnostic microarray. First, a multiplex PCR was developed to amplify all the DNA target sequences simultaneously. Second, the LDR-UA assay was implemented. The method was validated by analyzing 100 Italian Friesian DNA samples, which were also genotyped by conventional methods both at the protein level by means of milk isoelectrofocusing and at the molecular level using PCR-RFLP and PCR-single strand conformation polymorphism techniques. The genotypes obtained using the LDR-UA approach were in full agreement with those obtained by the conventional analyses. An important result of the LDR-UA assay was a more accurate genotyping of the different milk protein alleles than was found with conventional typing methods. At the kappa-casein gene, in fact, 4 samples were heterozygous (3 reference samples and 1 validation sample) for an allele coding for Thr(136) and Ala(148). This variant, which can be considered as the wild type of the genus Bos, is not usually identifiable by the conventional typing methods used. The multiplex PCR-LDR-UA approach developed provides for an accurate, inexpensive, and high-throughput assay that does not exhibit false positive or false negative signals, thus making it highly suitable for animal genotyping.

  11. Occurrence of flunixin residues in bovine milk samples from the USA.

    Science.gov (United States)

    Kissell, L W; Baynes, R E; Riviere, J E; Smith, G W

    2013-01-01

    5-Hydroxy-flunixin concentrations in milk samples were quantified by two commercially available screening assays--CHARM® and enzyme-linked immunoabsorbant assay (ELISA)--to determine whether any concentrations could be detected above the tolerance limit of 2 ng g⁻¹ from different regions in the United States. Milk samples came from large tanker trucks hauling milk to processing plants, and had already been screened for antibiotics. Positive results for flunixin residues based on a screening assay were confirmed by ultra-HPLC with mass spectrometric detection. Of the 500 milk samples analysed in this study, one sample was found to have a 5-hydroxy-flunixin concentration greater than the tolerance limit. The results of this study indicate that flunixin residues in milk are possible. Regulatory agencies should be aware that such residues can occur, and should consider incorporating or expanding flunixin screening tests as part of routine drug monitoring in milk. Larger studies are needed to determine the true prevalence of flunixin residues in milk from other regions in the United States as well as different countries.

  12. Interleukin-6 in quarter milk as a further prediction marker for bovine subclinical mastitis.

    Science.gov (United States)

    Sakemi, Yoko; Tamura, Yutaka; Hagiwara, Katsuro

    2011-02-01

    Subclinical mastitis (SM) is a major concern in the dairy industry because it causes economic losses and can lead to clinical mastitis. The mechanisms of the onset and progression of SM are not fully understood, and a new procedure for the detection and appropriate prediction of SM leading to clinical mastitis is necessary for dairy cow management. Inflammatory cytokines such as interleukin (IL)-6 are known to be present in inflamed mammary glands at the onset of mastitis, and IL-6 concentrations correlate with the levels of inflammation. In this study, the detection of IL-6 was examined for the evaluation for the future prediction of SM in 77 quarter milk samples from 20 cows. IL-6 concentrations in quarter milk were measured by sandwich ELISA, and the data were compared with milk somatic cell count (SCC) levels to diagnose SM. Average IL-6 concentration was significantly higher in SM quarter milk (207·0 ± 441·6 pg/ml) than in healthy control quarter milk (12·6 ± 33·4 pg/ml, P<0·01). Results of the cross-tabulation table show that SM prediction accuracy based on IL-6 concentration was almost equal or superior to SM prediction accuracy based on SCC. The detection of IL-6 in milk indicated SM earlier than did the detection of elevated SCC. Thus, the detection of IL-6 in milk could be a future prediction marker for SM.

  13. Predicting within-herd prevalence of infection with bovine leukemia virus using bulk-tank milk antibody levels.

    Science.gov (United States)

    Nekouei, Omid; Stryhn, Henrik; VanLeeuwen, John; Kelton, David; Hanna, Paul; Keefe, Greg

    2015-11-01

    Enzootic bovine leukosis (EBL) is an economically important infection of dairy cattle caused by bovine leukemia virus (BLV). Estimating the prevalence of BLV within dairy herds is a fundamental step towards pursuing efficient control programs. The objectives of this study were: (1) to determine the prevalence of BLV infection at the herd level using a bulk-tank milk (BTM) antibody ELISA in the Maritime region of Canada (3 provinces); and (2) to develop appropriate statistical models for predicting within-herd prevalence of BLV infection using BTM antibody ELISA titers. During 2013, three monthly BTM samples were collected from all dairy farms in the Maritime region of Canada (n=623) and tested for BLV milk antibodies using a commercial indirect ELISA. Based on the mean of the 3 BTM titers, 15 strata of herds (5 per province) were defined. From each stratum, 6 herds were randomly selected for a total of 90 farms. Within every selected herd, an additional BTM sample was taken (round 4), approximately 2 months after the third round. On the same day of BTM sampling, all cows that contributed milk to the fourth BTM sample were individually tested for BLV milk antibodies (n=6111) to estimate the true within-herd prevalence for the 90 herds. The association between true within-herd prevalence of BLV and means of various combinations of the BTM titers was assessed using linear regression models, adjusting for the stratified random sampling design. Herd level prevalence of BLV in the region was 90.8%. In the individual testing, 30.4% of cows were positive. True within-herd prevalences ranged from 0 to 94%. All linear regression models were able to predict the true within-herd prevalence of BLV reasonably well (R(2)>0.69). Predictions from the models were particularly accurate for low-to-medium spectrums of the BTM titers. In general, as a greater number of the four repeated BTM titers were incorporated in the models, narrower confidence intervals around the prediction lines

  14. A comparison of the determination of the rennet coagulation properties of bovine milk

    Directory of Open Access Journals (Sweden)

    Roman Pytel

    2016-07-01

    Full Text Available The aim of the work was compared of two different methods (the visual method and the nephelo turbidimetry method for determination of rennet coagulation time. It was observed the effect of heat treatment of milk; types of rennet and addition of different amount of CaCl2 into the pasteurized milk. It was used two different chymosin rennet. For the visual method was milk sample (100 mL equilibrated at 35 °C, 1 mL of rennet was added into milk and was measured the time required for the first visible flakes (visual method. For the determination rennet coagulation time by nephelo-turbidimetry was removed part of milk with rennet and placed into nephelo-turbidimetry. Milk had a titratable acidity in the range from 6.5 to 7.0 °SH, average pH of milk was 6.68. Dry matter content was in range from 12.351 to 13.142%. The average content of protein by Kjeldahl was 3.14%, fat by Gerber 4.34%, lactose by polarimetry 4.68% and calcium content 1.1%. The pasteurized milk had the worst rennet coagulation time about 32 s compared to the raw milk. The difference coagulation time between milk with addition of 20 µL CaCl2 and 40 µL CaCl2 was in range 21 s to 26 s by visual method. The difference coagulation time between milk with addition of 20 µL CaCl2 and 40 µL CaCl2 was 15 s by nephelo-turbidimetry method. There occurred statistically non-significant differences in most of the measurements, comparing the visual and the nephelo-turbidimetric method. The heat treatment, addition of CaCl2 and using of different rennet had an influence on the curd category. It was obtained, that using nephelo-turbidimetry shown objective results for measuring the rennet coagulation time contrary the subjective visual method. Further, the results obtained by nephelo-turbidimetry are accurate and determined with the lower variation.

  15. Kinetics of local and systemic isoforms of serum amyloid A in bovine mastitic milk

    DEFF Research Database (Denmark)

    Jacobsen, Stine; Niewold, T.A.; Kornalijnslijper, E.

    2005-01-01

    The aim of the present study was to characterise the serum amyloid A (SAA) response to intramammary inoculation of Escherichia coli and to examine the distribution of hepatically and extrahepatically pruduced SAA isoforms in plasma and milk fra cows with mastitis.......The aim of the present study was to characterise the serum amyloid A (SAA) response to intramammary inoculation of Escherichia coli and to examine the distribution of hepatically and extrahepatically pruduced SAA isoforms in plasma and milk fra cows with mastitis....

  16. Analysis on codon usage frequency in bovine milk protein%牛乳蛋白的遗传密码子使用频率分析

    Institute of Scientific and Technical Information of China (English)

    刘晶; 于浩; 杨润军; 鲍永华; 李俊雅; 戴蕴平; 赵志辉

    2011-01-01

    研究牛乳蛋白不同氨基酸的密码子使用频率对利用乳腺生物反应器表达目标蛋白过程中密码子的优化具有重要意义.本试验运用ENBOSS的CHIPS和SMS中的密码子使用工具对7种牛乳蛋白编码的基因进行了分析,并将这7个编码序列拼接在一起进行了乳蛋白全基因组的密码子偏爱性研究,同时与绵羊、猪及小鼠的乳蛋白密码子偏爱性进行了比较.结果表明:牛乳蛋白的CHIPS分析Nc值为51.867,αs1-CN、αs2-2-CN、β-CN、κ-CN、α-LB、β-IG、LTF的Nc值分别为52.005、49.967、48.746、58.113、55.514、36.274、47.646,编码牛乳蛋白氨基酸的密码子出现频率较均一;牛乳蛋白V、A、R、I、T、L、Q、P等氨基酸的密码子使用频率与其他物种有较大差异.牛乳蛋白的密码予偏爱性与绵羊最为接近.%In order to analyze bovine milk protein codon preference,7 coding sequences and the whole genome of bovine milk proteins were analyzed by CHIPS of EMBOSS and codon usage of SMS. The results were compared with codon usage of sheep,pig,and mouse milk protein. The results showed that the Nc value of all milk proteins in bovine and αs1- CN,αs2- CN,β- CN,κ- CN,α-LB,β-LG,LTF are 52. 005,49. 967,48. 746,58. 113,55. 514,36. 274,47. 646, respectively. Alternative codons for V, A, R, I, T, L, Q, P amino acids in bovine milk protein are distinctly different comparing with those of other species. It could be concluded that Nc value demonstrated that the codon usage frequency of bovine milk protein was comparatively uniform. Codon preference of bovine milk protein was the most close to that of sheep.

  17. Genetic parameters of saturated and monounsaturated fatty acid content and the ratio of saturated to unsaturated fatty acids in bovine milk.

    Science.gov (United States)

    Soyeurt, H; Dardenne, P; Dehareng, F; Bastin, C; Gengler, N

    2008-09-01

    Fatty acid composition influences the nutritional quality of milk and the technological properties of butter. Using a prediction of fatty acid (FA) contents by mid-infrared (MIR) spectrometry, a large amount of data concerning the FA profile in bovine milk was collected. The large number of records permitted consideration of more complex models than those used in previous studies. The aim of the current study was to estimate the effects of season and stage of lactation as well as genetic parameters of saturated (SAT) and monounsaturated (MONO) fatty acid contents in bovine milk and milk fat, and the ratio of SAT to unsaturated fatty acids (UNSAT) that reflect the hardness of butter (SAT:UNSAT), using 7 multiple-trait, random-regression test-day models. The relationship between these FA traits with common production traits was also studied. The data set contained 100,841 test-day records of 11,626 Holstein primiparous cows. The seasonal effect was studied based on unadjusted means. These results confirmed that milk fat produced during spring and summer had greater UNSAT content compared with winter (63.13 vs. 68.94% of SAT in fat, on average). The effect of stage of lactation on FA profile was studied using the same methodology. Holstein cows in early first lactation produced a lower content of SAT in their milk fat. Variance components were estimated using a Bayesian method via Gibbs sampling. Heritability of SAT in milk (0.42) was greater than heritability of SAT in milk fat (0.24). Estimates of heritability for MONO were also different in milk and fat (0.14 vs. 0.27). Heritability of SAT:UNSAT was moderate (0.27). For all of these traits, the heritability estimates and the genetic and phenotypic correlations varied through the lactation.

  18. Whole genome scan to detect quantitative trait loci for bovine milk protein composition.

    Science.gov (United States)

    Schopen, G C B; Koks, P D; van Arendonk, J A M; Bovenhuis, H; Visker, M H P W

    2009-08-01

    The objective of this study was to perform a whole genome scan to detect quantitative trait loci (QTL) for milk protein composition in 849 Holstein-Friesian cows originating from seven sires. One morning milk sample was analysed for the major milk proteins using capillary zone electrophoresis. A genetic map was constructed with 1341 single nucleotide polymorphisms, covering 2829 centimorgans (cM) and 95% of the cattle genome. The chromosomal regions most significantly related to milk protein composition (P(genome) casein, alpha(S2)-casein, beta-casein and kappa-casein. The QTL on BTA11 was found at 124 cM, and affected beta-lactoglobulin, and the QTL on BTA14 was found at 0 cM, and affected protein percentage. The proportion of phenotypic variance explained by the QTL was 3.6% for beta-casein and 7.9% for kappa-casein on BTA6, 28.3% for beta-lactoglobulin on BTA11, and 8.6% for protein percentage on BTA14. The QTL affecting alpha(S2)-casein on BTA6 and 17 showed a significant interaction. We investigated the extent to which the detected QTL affecting milk protein composition could be explained by known polymorphisms in beta-casein, kappa-casein, beta-lactoglobulin and DGAT1 genes. Correction for these polymorphisms decreased the proportion of phenotypic variance explained by the QTL previously found on BTA6, 11 and 14. Thus, several significant QTL affecting milk protein composition were found, of which some QTL could partially be explained by polymorphisms in milk protein genes.

  19. Polymorphisms in bovine immune genes and their associations with somatic cell count and milk production in dairy cattle

    Directory of Open Access Journals (Sweden)

    Magee David A

    2010-11-01

    Full Text Available Abstract Background Mastitis, an inflammation of the mammary gland, is a major source of economic loss on dairy farms. The aim of this study was to quantify the associations between two previously identified polymorphisms in the bovine toll-like receptor 2 (TLR2 and chemokine receptor 1 (CXCR1 genes and mammary health indictor traits in (a 246 lactating dairy cow contemporaries representing five breeds from one research farm and (b 848 Holstein-Friesian bulls that represent a large proportion of the Irish dairy germplasm. To expand the study, a further 14 polymorphisms in immune genes were included for association studies in the bull population. Results TLR4-2021 associated (P SERPINA1 haplotype with superior genetic merit for milk protein yield and milk fat percentage (P Conclusion Of the sixteen polymorphisms in seven immune genes genotyped, just CXCR1-777 tended to associate with SCS, albeit only in the on-farm study. The lack of an association between the polymorphisms with SCS in the Holstein-Friesian data set would question the potential importance of these variants in selection for improved mastitis resistance in the Holstein-Friesian cow.

  20. Epidemiological characteristics of bovine herpesvirus 1 infections determined by bulk milk testing of all Dutch dairy herds.

    Science.gov (United States)

    Van Wuijckhuise, L; Bosch, J; Franken, P; Frankena, K; Elbers, A R

    1998-02-21

    Samples of bulk milk were taken from all 33,636 Dutch dairy herds in November 1994 and tested for the presence of bovine herpesvirus 1 (BHV-1) antibodies with a gB-blocking ELISA. Sixteen per cent of the herds had a negative BHV-1 status in the bulk milk. Farms with only dairy cows were 1.9 times more likely to have a negative or weakly positive BHV-1 status than herds which also had beef/veal animals. Farms in areas containing less than one herd/km2 were 1.5 times more likely to have a negative or weakly positive BHV-1 status than herds in areas with more than three herds/km2. Differences in numbers of animals per unit area were not significantly associated with BHV-1 status. The probability of herds having a negative or weakly positive BHV-1 status decreased linearly with herd size by a factor of 1.2 per 10 animals. The purchase of stock was significantly associated with a negative or weakly positive BHV-1 status, but there was an interaction between farm type and purchase of stock. For farms with both dairy and beef/veal animals there was a weak association between the purchase of stock and BHV-1 status. For pure dairy herds the probability of having a negative or weakly positive BHV-1 status decreased linearly with the numbers of purchased stock by a factor of 1.3 per 10 animals purchased.

  1. Bovine milk oligosaccharides decrease gut permeability and improve inflammation and microbial dysbiosis in diet-induced obese mice.

    Science.gov (United States)

    Boudry, Gaëlle; Hamilton, M Kristina; Chichlowski, Maciej; Wickramasinghe, Saumya; Barile, Daniela; Kalanetra, Karen M; Mills, David A; Raybould, Helen E

    2017-01-25

    Obesity is characterized by altered gut homeostasis, including dysbiosis and increased gut permeability closely linked to the development of metabolic disorders. Milk oligosaccharides are complex sugars that selectively enhance the growth of specific beneficial bacteria in the gastrointestinal tract and could be used as prebiotics. The aim of the study was to demonstrate the effects of bovine milk oligosaccharides (BMO) and Bifidobacterium longum ssp. infantis (B. infantis) on restoring diet-induced obesity intestinal microbiota and barrier function defects in mice. Male C57/BL6 mice were fed a Western diet (WD, 40% fat/kcal) or normal chow (C, 14% fat/kcal) for 7 wk. During the final 2 wk of the study, the diet of a subgroup of WD-fed mice was supplemented with BMO (7% wt/wt). Weekly gavage of B. infantis was performed in all mice starting at wk 3, yet B. infantis could not be detected in any luminal contents when mice were killed. Supplementation of the WD with BMO normalized the cecal and colonic microbiota with increased abundance of Lactobacillus compared with both WD and C mice and restoration of Allobaculum and Ruminococcus levels to that of C mice. The BMO supplementation reduced WD-induced increase in paracellular and transcellular flux in the large intestine as well as mRNA levels of the inflammatory marker tumor necrosis factor α. In conclusion, BMO are promising prebiotics to modulate gut microbiota and intestinal barrier function for enhanced health.

  2. Occurrence of enterotoxin genes and macrorestriction analysis of Staphylococcus aureus isolated from bovine mastitis and bulk-tank milk samples in Italy. An epidemiological study

    Directory of Open Access Journals (Sweden)

    Roberto Rosmini

    2010-01-01

    Full Text Available The goal of the study was to genotypically compare S. aureus isolates from mastitis milk and raw milk to identify therelation between strains and to assess the enterotoxigenicity of the isolates. Eighty-three Staphylococcus aureus isolatesrecovered from cows and bulk tank milk of five farms in northern Italy were compared genotypically. The genes for theenterotoxins A, D, G and I, but not for B, C, E and H and the toxic shock syndrome toxin 1 (TSST-1, were detected byPCR amplification. Macrorestriction analysis with the restrictions enzyme SmaI revealed 14 pulsed-field gel electrophoresispatterns. These were in part different from each other only in a few fragments and thus displayed a closeclonal relation. The results of the present investigation showed that identical or closely related clones seemed to beresponsible for the cases of bovine mastitis in the farms investigated and partly responsible for contamination of bulktank milk.

  3. The association of low-molecular-weight hydrophobic compounds with native casein micelles in bovine milk.

    Science.gov (United States)

    Cheema, M; Mohan, M S; Campagna, S R; Jurat-Fuentes, J L; Harte, F M

    2015-08-01

    The agreed biological function of the casein micelles in milk is to carry minerals (calcium, magnesium, and phosphorus) from mother to young along with amino acids for growth and development. Recently, native and modified casein micelles were used as encapsulating and delivery agents for various hydrophobic low-molecular-weight probes. The ability of modified casein micelles to bind certain probes may derive from the binding affinity of native casein micelles. Hence, a study with milk from single cows was conducted to further elucidate the association of hydrophobic molecules into native casein micelles and further understand their biological function. Hydrophobic and hydrophilic extraction followed by ultraperformance liquid chromatography-high resolution mass spectrometry analysis were performed over protein fractions obtained from size exclusion fractionation of raw skim milk. Hydrophobic compounds, including phosphatidylcholine, lyso-phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin, showed strong association exclusively to casein micelles as compared with whey proteins, whereas hydrophilic compounds did not display any preference for their association among milk proteins. Further analysis using liquid chromatography-tandem mass spectrometry detected 42 compounds associated solely with the casein-micelles fraction. Mass fragments in tandem mass spectrometry identified 4 of these compounds as phosphatidylcholine with fatty acid composition of 16:0/18:1, 14:0/16:0, 16:0/16:0, and 18:1/18:0. These results support that transporting low-molecular-weight hydrophobic molecules is also a biological function of the casein micelles in milk.

  4. Fatty Acid Content of Bovine Milkfat From Raw Milk to Yoghurt

    Directory of Open Access Journals (Sweden)

    O. O. Santos Junior

    2012-01-01

    Full Text Available Problem statement: The present study aimed to study the evolution of fatty acid content, focusing on rumenic acid content, from raw milk to yoghurt processed from this milk. Approach: Milk samples were collected in a dairy plant in the northwest of Parana State weekly in January 2011 (Brazilian summer. It processed one truck load of 26,000 L of refrigerated type-C (whole standardized milk with a minimum of 3% fat milk per day, mostly from the city of Lobato, Parana, produced mainly by Gir (Bos indicus cattle raised on stargrass (Cynodon nlenfuensis var. nlenfuensis pasture. Results: Saturated Fatty Acid (SFA were the most abundant, particularly palmitic (16:0, stearic (18:0 and myristic (14:0. Among the Monounsaturated Fatty Acid (MUFA, Polyunsaturated Fatty Acid (PUFA and trans fatty acid, oleic acid (18:1n-9, linoleic acid (18:2n-6, elaidic acid (t9-18:1 and c9, t11-18:2 (rumenic acid predominated. It was detected significant differences (p-1 of lipids, decreasing to 6.22±0.20 after pasteurization and to 5.41±0.18 mg g-1 in yoghurt. Conclusion/Recommendations: It is demonstrated that pasteurization and yoghurt making affect the CLA contents.

  5. 母乳和牛乳中乳脂肪球膜蛋白质的差异分析%Analysis of Differences in Milk Fat Globule Membrane Proteins between Breast Milk and Bovine Milk

    Institute of Scientific and Technical Information of China (English)

    景萌娜; 姜铁民; 刘斌; 张咚咚; 魏京华; 王品; 李菊芳; 陈历俊

    2016-01-01

    In order to compare the difference in milk fat globule membrane (MFGM) proteins between breast milk and bovine milk, MFGM were extracted from milk by organic solvent extraction and lysed in SDS-containing lysis buffer for analysis by liquid chromatography and mass spectrometry (LC-MS) after the removal of SDS using PierceTM detergent removal spin column. The results were filtered according to the number of unique peptide≥ 1. A total of 863 and 454 proteins were identiifed separately in breast milk and bovine milk, including 175 proteins in common, and 688 and 279 proteins unique for breast milk and bovine milk, respectively. Gene Ontology (GO) analysis of 688 speciifc MFGM proteins in human milk showed that they were mainly distributed in cell part, cell organelle, and cell membrane, and involved in biological pathways such as metabolic processes, biological regulation, response to stimulus, immune system process, reproduction, and so on. These proteins also participated in many molecular functions, such as catalytic activity, binding, transport activity and enzyme regulation activity. In addition, these breast milk MFGM proteins were involved in some metabolic pathways, such as aminoacyl-tRNA biosynthesis, pathogenicEscherichia coli infection, fatty acid metabolism, propanoate metabolism, steroid biosynthesis, lysine degradation, which were not shown in bovine milk. The MFGM proteins in breast milk and bovine milk had obvious differences. Although some compositions and functions were overlapped, in terms of abundance and metabolic pathways, the MFGM proteins in bovine milk could not be replaced by those in breast milk.%为比较乳脂肪球膜(milk fat globule membrane,MFGM)蛋白在母乳和牛乳中的差异,采用有机试剂提取法,从母乳和牛乳中提取分离出MFGM蛋白,经皮尔斯TM去垢小柱去除

  6. Mid-infrared prediction of bovine milk fatty acids across multiple breeds, production systems, and countries.

    Science.gov (United States)

    Soyeurt, H; Dehareng, F; Gengler, N; McParland, S; Wall, E; Berry, D P; Coffey, M; Dardenne, P

    2011-04-01

    Increasing consumer concern exists over the relationship between food composition and human health. Because of the known effects of fatty acids on human health, the development of a quick, inexpensive, and accurate method to directly quantify the fatty acid (FA) composition in milk would be valuable for milk processors to develop a payment system for milk pertinent to their customer requirements and for farmers to adapt their feeding systems and breeding strategies accordingly. The aim of this study was (1) to confirm the ability of mid-infrared spectrometry (MIR) to quantify individual FA content in milk by using an innovative procedure of sampling (i.e., samples were collected from cows belonging to different breeds, different countries, and in different production systems); (2) to compare 6 mathematical methods to develop robust calibration equations for predicting the contents of individual FA in milk; and (3) to test interest in using the FA equations developed in milk as basis to predict FA content in fat without corrections for the slope and the bias of the developed equations. In total, 517 samples selected based on their spectral variability in 3 countries (Belgium, Ireland, and United Kingdom) from various breeds, cows, and production systems were analyzed by gas chromatography (GC). The samples presenting the largest spectral variability were used to calibrate the prediction of FA by MIR. The remaining samples were used to externally validate the 28 FA equations developed. The 6 methods were (1) partial least squares regression (PLS); (2) PLS+repeatability file (REP); (3) first derivative of spectral data+PLS; (4) first derivative+REP+PLS; (5) second derivative of spectral data+PLS; and (6) second derivative+REP+PLS. Methods were compared on the basis of the cross-validation coefficient of determination (R2cv), the ratio of standard deviation of GC values to the standard error of cross-validation (RPD), and the validation coefficient of determination (R2

  7. Arsenic concentration in water and bovine milk in Cordoba, Argentina. Preliminary results.

    Science.gov (United States)

    Pérez-Carrera, Alejo; Fernández-Cirelli, Alicia

    2005-02-01

    The Chaco Pampean Plain of central Argentina constitutes one of the largest regions of high arsenic (As) groundwaters known, covering around 1 x 10(6) km2 (Smedley & Kinniburg, 2002; Farías et al. 2004). The high-As groundwaters are from Quaternary deposits of loess (mainly silt) with intermixed rhyolitic or dacitic volcanic ash (Nicolli et al. 1989, Smedley et al. 1998,2002). Early in the last century an endemic disease due to contamination of drinking water with arsenic was recognised. This disease is called HACRE (Hidroarsenicismo Crónico Regional Endémico, Chronic Endemic Regional Hydroarsenism) and is connected with a particular type of skin cancer (Astolfi et al. 1981). One of the most affected region is the province of Cordoba, where Nicolli et al. (1989) reported As concentrations that exceed the maximun level permitted for drinking water of 50 microg/l for 82% of the groundwater samples (n=60) of a study area comprising approximately 10000 km2. The southeast of Cordoba is an important milk production zone in Argentina, where dairy product consumption is up to 192 equivalent milk l/inhabitant/year. As a secretion of the mammary gland, milk can carry numerous xenobiotic substances, which constitute a technological risk factor for dairy products and above all for the health of the consumer (Licata et al. 2004). Nevertheless no studies on the incidence of high-As livestock drinking water in livestock health and its transfer to milk have been performed in Argentina. The aim of the present study was the determination of arsenic content in livestock drinking water and milk from dairy farms located in an area of high-As groundwaters, to analyse the relation between As uptake through water and its transfer to milk.

  8. Prediction of bovine milk true protein content by mid-infrared spectroscopy

    OpenAIRE

    Botaro,Bruno Garcia; CORTINHAS, Cristina Simões; Mestieri, Lucinéia; Machado,Paulo Fernando; Santos, Marcos Veiga dos

    2011-01-01

    The aim of this study was to estimate the concentration of milk true protein (TP) by mid-infrared absorbance method (MIR) in samples from bulk tank of dairy herds, and to determine the correlation between the results of TP of milk determined by Kjeldahl and MIR. Forty nine dairy herds were selected (17 Holstein, 6 Jersey and 26 Girolando) for monthly collections of samples from bulk tanks during the period of one year (284 samples). Fat, lactose, crude protein and total solids were firstly de...

  9. Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    OpenAIRE

    Xia, Xiaojing; Che, Yanyi; Gao, Yuanyuan; Zhao, Shuang; Ao, Changjin; Yang, Hongjian; Liu, Juxiong; Liu, Guowen; Han, Wenyu; Wang, Yuping; Lei, Liancheng

    2016-01-01

    During the lactation cycle of the bovine mammary gland, autophagy is induced in bovine mammary epithelial cells (BMECs) as a cellular homeostasis and survival mechanism. Interferon gamma (IFN-γ) is an important antiproliferative and apoptogenic factor that has been shown to induce autophagy in multiple cell lines in vitro. However, it remains unclear whether IFN-γ can induce autophagy and whether autophagy affects milk synthesis in BMECs. To understand whether IFN-γ affects milk synthesis, we...

  10. Bovine colostrum improves neonatal growth, digestive function, and gut immunity relative to donor human milk and infant formula in preterm pigs

    DEFF Research Database (Denmark)

    Rasmussen, Stine Ostenfeldt; Martin, Lena; Østergaard, Mette Viberg;

    2016-01-01

    Mother's own milk is the optimal first diet for preterm infants, but donor human milk (DM) or infant formula (IF) is used when supply is limited. We hypothesized that a gradual introduction of bovine colostrum (BC) or DM improves gut maturation, relative to IF during the first 11 days after preterm...... DM and BC pigs. BC, and to some degree DM, are superior to preterm IF in stimulating gut maturation and body growth, using a gradual advancement of enteral feeding volume over the first 11 days after preterm birth in piglets. Whether the same is true in preterm infants remains to be tested....

  11. Proteomic analysis of differentially expressed proteins in bovine milk during experimentally induced Escherichia coli mastitis

    Science.gov (United States)

    The objectives of the current study were to profile changes in protein composition using 2-dimensional gel electrophoresis (2D-GE) on whey samples from a group of 8 cows prior to and 18 hours after infection with Escherichia coli, and to identify differentially expressed milk proteins by peptide seq...

  12. Bovine chromosomal regions affecting rheological traits in rennet-induced skim milk gels

    DEFF Research Database (Denmark)

    Gregersen, Vivi Raundahl; Gustavsson, F; Glantz, M

    2015-01-01

    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...

  13. Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine milk samples

    NARCIS (Netherlands)

    Sampimon, O.C.; Lam, T.G.J.M.; Mevius, D.J.; Schukken, Y.H.; Zadoks, R.N.

    2011-01-01

    The aim of this study was to examine whether antimicrobial resistance profiles of coagulase-negative Staphylococcus (CNS) species isolated from milk of dairy cows differed between bacterial species, and to compare results obtained by phenotypic and genotypic profiling of resistance to penicillin, ox

  14. High concentration of human lactoferrin in milk of rhLf-transgenic cows relieves signs of bovine experimental Staphylococcus chromogenes intramammary infection.

    Science.gov (United States)

    Simojoki, Heli; Hyvönen, Paula; Orro, Toomas; Pyörälä, Satu

    2010-08-15

    Six transgenic cows producing recombinant human lactoferrin (rhLf) in their milk and five normal cows at the same lactation stage were experimentally infected with Staphylococcus chromogenes to study the effect of a high concentration of lactoferrin in milk. Coagulase-negative staphylococci such as S. chromogenes have become very common as agents causing mild or subclinical mastitis. All transgenic cows became infected but showed no clinical signs, unlike the control cows, which developed mild clinical mastitis. Transgenic cows eliminated bacteria faster from the quarters than did the controls. Local clinical signs were milder, and the inflammatory reaction assessed by NAGase activity in the milk and by the concentration of milk amyloid A was lower in the transgenic cows. The mild response probably reflected the rapid elimination of bacteria. The milk concentration of rhLf remained constant throughout the study period, but the total concentration of bovine lactoferrin in the milk peaked in both groups at 46h post-challenge. Three cows, all in the control group, exhibited systemic acute phase response as increased concentrations of serum amyloid A in the blood circulation. Transgenic cows with a high concentration of human lactoferrin in their milk seemed to be protected from clinical disease and from prolonged inflammatory reaction, but not from experimental intramammary infection induced by S. chromogenes.

  15. KONSENTRASI MINIMUM SISTEM LAKTOPEROKSIDASE UNTUK MENEKAN PERTUMBUHAN Escherichia coli PADA SUSU SAPI SEGAR [Minimum Inhibitory Concentration of Lactoperoxidase System Against Escherichia coli in Fresh Bovine Milk

    Directory of Open Access Journals (Sweden)

    Bagus Fitriansyah

    2015-07-01

    Full Text Available Fresh milk production in Central Java has increased every year resulting in the requirement of the rapid and good process for fresh milk handling to keep the quality against bacteria. Lactoperoxidase system (LPOS was well documented as a strategy to extend the shelf life of fresh milk but it hasn’t been known yet for the minimum level of lactoperoxidase (LPO utilization to activate LPOS in the milk. The purpose of this study was to analyze minimum concentration of LPO and its components by calculating minimum inhibitory concentration (MIC against Escherichia coli and total bacteria in fresh milk. LPO was obtained from bovine whey and its purity was analyzed using SDS-PAGE. To activate the LPO performance, two substates namely H2O2 and KSCN were used with various concentrations. The MIC was analyzed 3 times using 3M™ Petrifilm™ Aerobic Count Plates. The result showed that 3.11 unit/mL LPO, 0.03 mM subs-trates have been found as MIC of LPOS against bacteria and Escherichia coli in milk. This result might provide benefit to the utilization of minimal LPOS in fresh milk to safe the amount of LPO.

  16. Hot topic: Bovine milk samples yielding negative or nonspecific results in bacterial culturing--the possible role of PCR-single strand conformation polymorphism in mastitis diagnosis.

    Science.gov (United States)

    Schwaiger, K; Wimmer, M; Huber-Schlenstedt, R; Fehlings, K; Hölzel, C S; Bauer, J

    2012-01-01

    A large proportion of mastitis milk samples yield negative or nonspecific results (i.e., no mastitis pathogen can be identified) in bacterial culturing. Therefore, the culture-independent PCR-single strand conformation polymorphism method was applied to the investigation of bovine mastitis milk samples. In addition to the known mastitis pathogens, the method was suitable for the detection of fastidious bacteria such as Mycoplasma spp., which are often missed by conventional culturing methods. The detection of Helcococcus ovis in 4 samples might indicate an involvement of this species in pathogenesis of bovine mastitis. In conclusion, PCR-single-strand conformation polymorphism is a promising tool for gaining new insights into the bacteriological etiology of mastitis.

  17. Optimization and characterization of an in vitro bovine mammary cell culture system to study regulation of milk protein synthesis and mammary differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Talhouk, R.S.

    1988-01-01

    A long term bovine mammary cell culture system that maintains normal mammary cell function was established and optimized to study milk protein synthesis and secretion and mammary differentiation. This culture system used bovine mammary acini isolated from developing or lactating mammary gland by enzymatic dissociation, and cryopreserved until thawed and plated for growth in vitro for these studies. Cells in M199 with lactogenic hormones {plus minus} fetal calf serum (FCS) were cultured on plastic, 100ul and 500ul type I collagen, and Matrigel, or embedded within type I collagen. Cell morphology, cell number, and total TCA-precipitable {sup 35}S-labelled proteins were monitored. Milk protein ({alpha}{sub s,1}-casein, lactoferrin (LF), {alpha}-lactalbumin, and {beta}-lactoglobulin) secretion and intracellular levels were determined by an ELISA assay.

  18. Application of Proteomics in Bovine Milk Research%蛋白质组学技术在乳品研究中的应用

    Institute of Scientific and Technical Information of China (English)

    芦晶; 刘鹭; 张书文; 吕加平

    2013-01-01

    Milk is an important nutritional food. Identification and application of its nutritional composition has been plagued by methodological limitations. With the development of proteomics technology, we have deeper understanding on protein composition and properties of milk. This paper introduces the steps of proteomics technology, reviews the research progress and applications in the bovine milk research, including casein proteomics, whey protein proteomics and milk fat globule membrane protein proteomics. It will help to fully understand the mechanism of milk synthesis and secretion, increase the milk production and utilization of bovine milk as source of nutrients.%牛乳是重要的营养食品,其营养组分的鉴定和应用已受到传统方法的限制。通过蛋白质组学技术的发展,对牛乳中蛋白质的组成和性质有了更深入的认识。本文综述了蛋白质组学技术的步骤及其在乳品研究中的应用进展,包括酪蛋白蛋白质组学、乳清蛋白质组学和乳脂肪球膜蛋白质组学的研究。为充分理解牛乳分泌机理,有效控制牛乳生产,提高牛乳营养价值及利用提供理论依据。

  19. Prediction of bovine milk technological traits from mid-infrared spectroscopy analysis in dairy cows.

    Science.gov (United States)

    Visentin, G; McDermott, A; McParland, S; Berry, D P; Kenny, O A; Brodkorb, A; Fenelon, M A; De Marchi, M

    2015-09-01

    Rapid, cost-effective monitoring of milk technological traits is a significant challenge for dairy industries specialized in cheese manufacturing. The objective of the present study was to investigate the ability of mid-infrared spectroscopy to predict rennet coagulation time, curd-firming time, curd firmness at 30 and 60min after rennet addition, heat coagulation time, casein micelle size, and pH in cow milk samples, and to quantify associations between these milk technological traits and conventional milk quality traits. Samples (n=713) were collected from 605 cows from multiple herds; the samples represented multiple breeds, stages of lactation, parities, and milking times. Reference analyses were undertaken in accordance with standardized methods, and mid-infrared spectra in the range of 900 to 5,000cm(-1) were available for all samples. Prediction models were developed using partial least squares regression, and prediction accuracy was based on both cross and external validation. The proportion of variance explained by the prediction models in external validation was greatest for pH (71%), followed by rennet coagulation time (55%) and milk heat coagulation time (46%). Models to predict curd firmness 60min from rennet addition and casein micelle size, however, were poor, explaining only 25 and 13%, respectively, of the total variance in each trait within external validation. On average, all prediction models tended to be unbiased. The linear regression coefficient of the reference value on the predicted value varied from 0.17 (casein micelle size regression model) to 0.83 (pH regression model) but all differed from 1. The ratio performance deviation of 1.07 (casein micelle size prediction model) to 1.79 (pH prediction model) for all prediction models in the external validation was <2, suggesting that none of the prediction models could be used for analytical purposes. With the exception of casein micelle size and curd firmness at 60min after rennet addition

  20. Development and validation of an ultra high performance liquid chromatography tandem mass spectrometry method for simultaneous determination of sulfonamides, quinolones and benzimidazoles in bovine milk.

    Science.gov (United States)

    Hou, Xiao-Lin; Chen, Guo; Zhu, Li; Yang, Ting; Zhao, Jian; Wang, Lei; Wu, Yin-Liang

    2014-07-01

    A simple, sensitive and reliable analytical method was developed for the simultaneous determination of 38 veterinary drugs (18 sulfonamides, 11 quinolones and 9 benzimidazoles) and 8 metabolites of benzimidazoles in bovine milk by ultra high performance liquid chromatography-positive electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS). Samples were extracted with acidified acetonitrile, cleaned up with Oasis(®) MCX cartridges, and analyzed by LC-MS/MS on an Acquity UPLC(®) BEH C18 column with gradient elution. The method allows such multi-analyte measurements within a 13min runtime while the specificity is ensured through the MRM acquisition mode. The method was validated according to the European Commission Decision 2002/657/EC determining specificity, decision limit (CCα), detection capability (CCβ), recovery, precision, linearity and stability. For compounds which have MRLs in bovine milk, the CCα values fall into a range from 11 to 115μg/kg, and the CCβ values fall within a range of 12-125μg/kg. For compounds which have not MRLs in bovine milk, the CCα values fall into a range from 0.01 to 0.08μg/kg, and the CCβ values fall within a range of 0.02-0.11μg/kg. The mean recoveries of the 46 analytes were between 87 and 119%. The calculated RSD values of repeatability and within-laboratory reproducibility experiments were below 11% and 15% for the 46 compounds, respectively. The method was demonstrated to be suitable for the simultaneous determination of sulfonamides, quinolones and benzimidazoles in bovine milk.

  1. Prevalence of Bovine Brucellosis in Organized Dairy Farms, Using Milk ELISA, in Quetta City, Balochistan, Pakistan

    Directory of Open Access Journals (Sweden)

    Muhammad Shafee

    2011-01-01

    Full Text Available A total of 200 milk samples from cattle (=86 and buffalo (=114 were evaluated using milk ring test (MRT and indirect enzyme-linked immunosorbent assay (i-ELISA. The overall prevalence was found to be 3% and 8.5% in cattle and buffaloes using MRT and i-ELISA, respectively. The prevalence was 4.6% and 1.7% in cattle and buffalo using MRT, respectively, while i-ELISA exhibited 20% and 0% in cattle and buffalo, respectively. The prevalence was higher in government dairy farm, compared to privately owned dairy farm. This paper points out an alarming situation in the target area with respect to the public health significance.

  2. Prevalence of bovine brucellosis in organized dairy farms, using milk ELISA, in quetta city, balochistan, pakistan.

    Science.gov (United States)

    Shafee, Muhammad; Rabbani, Masood; Sheikh, Ali Ahmad; Ahmad, Mansoor Din; Razzaq, Abdul

    2011-01-24

    A total of 200 milk samples from cattle (n = 86) and buffalo (n = 114) were evaluated using milk ring test (MRT) and indirect enzyme-linked immunosorbent assay (i-ELISA). The overall prevalence was found to be 3% and 8.5% in cattle and buffaloes using MRT and i-ELISA, respectively. The prevalence was 4.6% and 1.7% in cattle and buffalo using MRT, respectively, while i-ELISA exhibited 20% and 0% in cattle and buffalo, respectively. The prevalence was higher in government dairy farm, compared to privately owned dairy farm. This paper points out an alarming situation in the target area with respect to the public health significance.

  3. Simultaneous determination of β-lactam antibiotics and β-lactamase inhibitors in bovine milk by ultra performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Li, Nasi; Feng, Feng; Yang, Bingcheng; Jiang, Pingping; Chu, Xiaogang

    2014-01-15

    An ultra performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method has been developed for the simultaneous determination of four β-lactam antibiotics (amoxicillin, ampicillin, cefotaxime, and cefoperazone) and two β-lactamase inhibitors (tazobactam, sulbactam) in bovine milk. The analytes were extracted with water from bovine milk and purified with Oasis HLB solid phase extraction (SPE) cartridges. The analytes were determined in less than 3min by UPLC-MS/MS in positive and negative electrospray ionization (ESI) modes, separately. The method was linear over the range of 1-100μg/L for tazobactam, sulbactam, ampicillin, and cefoperazone, and 2-100μg/L for amoxicillin and cefotaxime. The recoveries for all six analytes in bovine milk ranged from 82.5 to 98.3%. The limits of detection and the limits of quantitation were 0.1-0.2μg/L and 0.3-0.5μg/L, respectively. The intra- and inter-day precisions were less than 6% for each compound.

  4. Sensitivity and Selectivity on Aptamer-Based Assay: The Determination of Tetracycline Residue in Bovine Milk

    Directory of Open Access Journals (Sweden)

    Sohee Jeong

    2012-01-01

    Full Text Available A competitive enzyme-linked aptamer assay (ELAA to detect tetracycline in milk was performed by using two different aptamers individually; one is 76 mer-DNA aptamer and the other is 57 mer-RNA aptamer. The best optimum condition was obtained without monovalent ion, Na+ and also by adding no Mg2+ ion in the assay buffer, along with RT incubation. The optimized ELAA showed a good sensitivity (LOD of 2.10 × 10−8 M with a wide dynamic range (3.16 × 10−8 M ~ 3.16 × 10−4 M. In addition, the average R.S.D. across all data points of the curve was less than 2.5% with good recoveries (~101.8% from the milk media. Thus, this method provides a good tool to monitor tetracycline in milk from MRLs’ point of view. However, this ELAA method was not superior to the ELISA method in terms of specificity. This paper describes that it does not always give better sensitivity and specificity in assays even though aptamers have several advantages over antibodies and have been known to be good binders for binding assays.

  5. Rapid confirmatory analysis of non-steroidal anti-inflammatory drugs in bovine milk by rapid resolution liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Dowling, Geraldine; Gallo, Pasquale; Malone, Edward; Regan, Liam

    2009-11-13

    A rapid method has been developed to analyse carprofen (CPF), diclofenac (DCF), mefenamic acid (MFN), niflumic acid (NIFLU), naproxen (NAP), oxyphenylbutazone (OXYPHEN), phenylbutazone (PBZ) and suxibuzone (SUXI) residues in bovine milk. Milk samples are extracted with acetonitrile and sample extracts were purified on Evolute ABN solid phase extraction cartridges. Aliquots were analysed by rapid resolution liquid chromatography tandem mass spectrometry (RRLC-MS/MS) with a runtime of 6.5 min. The method was validated in bovine milk, according to the criteria defined in Commission Decision 2002/657/EC. CCalpha values of 0.46, 1.08, 0.92, 1.26, 1.29, 2.12, 0.55 and 2.86 ng mL(-1) were determined for CPF, DCF, MFN, NIFLU, NAP, OXYPHEN, PBZ and SUXI, respectively. CCbeta values of 0.79, 1.85, 1.56, 2.15, 2.19, 3.62, 0.94 and 4.87 ng mL(-1) were determined for CPF, DCF, MFN, NIFLU, NAP, OXYPHEN, PBZ and SUXI, respectively. The measurement uncertainty of the method was estimated at 9, 28, 28, 45, 46, 45, 10 and 39% for CPF, DCF, MFN, NIFLU, NAP, OXYPHEN, PBZ and SUXI. Fortifying bovine milk samples (n=18) in three separate assays, show the accuracy of the method to be between 82 and 108%. The precision of the method, expressed as RSD values for the within-lab reproducibility at the three levels of fortification (5, 7.5 and 10 ng mL(-1)) was less than 16%, respectively. The advantage of the method is that low ng mL(-1) levels can be detected and quantitatively confirmed rapidly in milk and that 3 batches of samples can be analysed within a single day using RRLC-MS/MS with a runtime of 6.5 min.

  6. Features of the milk whey protein partitioning in polyethyleneglycol-sodium citrate aqueous two-phase systems with the goal of isolating human alpha-1 antitrypsin expressed in bovine milk.

    Science.gov (United States)

    Boaglio, Andrea; Bassani, Georgina; Picó, Guillermo; Nerli, Bibiana

    2006-06-06

    Partitioning behaviour of the bovine whey proteins (bovine serum albumin, alpha-lactoalbumin and beta-lactoglobulin) and human alpha-1 antitrypsin in aqueous two-phase systems prepared with polyethyleneglycol (molecular masses: 1000, 1450 and 3350)-sodium citrate was analysed at pH 5.2, 6.2 and 8.2. Alpha lactoalbumin concentrated in the polyethyleneglycol rich-phase, while beta-lactoglobulin, bovine serum albumin and alpha-1 antitrypsin showed affinity for the citrate rich-phase. In aqueous two-phase systems of high medium pH and high polyethyleneglycol molecular mass the protein partitioning equilibrium is displaced to the citrate rich-phase. The polyethyleneglycol 1450-pH 5.2 system with a top/bottom phase-volume ratio of 3 showed to have the best capability of recovering the alpha-1 antitrypsin from a mixture prepared with natural milk whey and human alpha-1 antitrypsin. The recovery of this protein in the bottom phase was of 90% and the purity of the obtained product was of 98%. The method appears to be suitable as a starting point to isolate other human proteins expressed in transgenic bovine milk.

  7. Camel and bovine chymosin

    DEFF Research Database (Denmark)

    Langholm Jensen, Jesper; Mølgaard, Anne; Navarro Poulsen, Jens Christian;

    2013-01-01

    Bovine and camel chymosin are aspartic peptidases that are used industrially in cheese production. They cleave the Phe105-Met106 bond of the milk protein κ-casein, releasing its predominantly negatively charged C-terminus, which leads to the separation of the milk into curds and whey. Despite...... having 85% sequence identity, camel chymosin shows a 70% higher milk-clotting activity than bovine chymosin towards bovine milk. The activities, structures, thermal stabilities and glycosylation patterns of bovine and camel chymosin obtained by fermentation in Aspergillus niger have been examined...... interactions arising from variation in the surface charges and the greater malleability both in domain movements and substrate binding contribute to the better milk-clotting activity of camel chymosin towards bovine milk....

  8. Expression of CD14 and toll-like receptors 2 and 4 by milk neutrophils in bovine mammary glands infected with Corynebacterium bovis

    Directory of Open Access Journals (Sweden)

    Maiara G. Blagitz

    2015-01-01

    Full Text Available This study evaluated the expression of CD14, toll-like receptor (TLR 2 and TLR4 on the surface of milk neutrophils in bovine mammary glands infected with Corynebacterium bovis. Here, we used 23 culture-negative control quarters with no abnormal secretion on the strip cup test and milk somatic cell count lower than 1x105 cells/mL, and 14 C. bovis infected quarters. The identification of neutrophils, as well as, the percentage of neutrophils that expressed CD14, TLR2 and TLR4 were analyzed by flow cytometry using monoclonal antibodies. The present study encountered no significant difference in the percentages of milk neutrophils that expressed TLR2 and TLR4 or in the expression of TLR4 by milk neutrophils. Conversely, a lower median fluorescence intensity of TLR2 in milk neutrophils was observed in C. bovis-infected quarters. The percentage of neutrophils that expressed CD14 and the median fluorescence intensity of CD14 in milk neutrophils was also lower in C. bovis-infected quarters.

  9. Simultaneous determination of nitroimidazoles, benzimidazoles, and chloramphenicol components in bovine milk by ultra-high performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Wang, Yuanyuan; Li, Xiaowei; Zhang, Zhiwen; Ding, Shuangyang; Jiang, Haiyang; Li, Jiancheng; Shen, Jianzhong; Xia, Xi

    2016-02-01

    A sensitive, confirmatory ultra-high performance liquid chromatography-tandem mass spectrometric method was developed and validated to detect 23 veterinary drugs and metabolites (nitroimidazoles, benzimidazoles, and chloramphenicol components) in bovine milk. Compounds of interest were sequentially extracted from milk with acetonitrile and basified acetonitrile using sodium chloride to induce liquid-liquid partition. The extract was purified on a mixed mode solid-phase extraction cartridge. Using rapid polarity switching in electrospray ionization, a single injection was capable of detecting both positively and negatively charged analytes in a 9 min chromatography run time. Recoveries based on matrix-matched calibrations and isotope labeled internal standards for milk ranged from 51.7% to 101.8%. The detection limits and quantitation limits of the analytical method were found to be within the range of 2-20 ng/kg and 5-50 ng/kg, respectively. The recommended method is simple, specific, and reliable for the routine monitoring of nitroimidazoles, benzimidazoles, and chloramphenicol components in bovine milk samples.

  10. Effects of Cooling and Supplemental Bovine Somatotropin on Milk Production relating to Body Glucose Metabolism and Utilization of Glucose by the Mammary Gland in Crossbred Holstein Cattle

    Directory of Open Access Journals (Sweden)

    Siravit Sitprija

    2010-01-01

    Full Text Available Problem statement: The low milk yield and shorter persistency of lactation of dairy cattle is the major problem for the dairy practices in the tropics. High environmental temperatures and rapid decline of plasma growth hormone level can influence milk production. Regulation of the milk yield of animals is mainly based on the mechanisms governing the quantity of glucose extracted by the mammary gland for lactose biosynthetic pathways. The mechanism(s underlying the effects of cooling and supplemental bovine somatotropin on milk production relating to body glucose metabolism and intracellular metabolism of glucose in the mammary gland of crossbred Holstein cattle in the tropics have not been investigated to date. Approach: Ten crossbred 87.5% Holstein cows were divided into two groups of five animals each. Animals were housed in Normal Shade barn (NS as non-cooled cows and cows in the second group were housed in barn which was equipped with a two Misty-Fan cooling system (MF as cooled cows. Supplementation of recombinant bovine Somatotropin (rbST (POSILAC, 500 mg per cow were performed in both groups to study body glucose metabolism and the utilization of glucose in the mammary gland using a continuous infusion of [3-3H] glucose and [U- 14C] glucose as markers in early, mid and late stages of lactation. Results: Milk yield significantly increased in both groups during supplemental rbST with a high level of mammary blood flow. Body glucose turnover rates were not significant different between cooled and non-cooled cows whether supplemental rbST or not. The glucose taken up by the mammary gland of both non-cooled and cooled cows increased flux through the lactose synthesis and the pentose cycle pathway with significant increases in NADPH formation for fatty acid synthesis during rbST supplementation. The utilization of glucose carbon incorporation into milk appeared to increase in milk lactose and milk triacylglycerol but not for

  11. Probability of detecting antibodies to bovine herpesvirus 1 in bulk milk after the introduction of a positive animal on to a negative farm.

    Science.gov (United States)

    Frankena, K; Franken, P; Vandehoek, J; Koskamp, G; Kramps, J A

    1997-01-25

    The purpose of this study was to assess the probability that the introduction of one or more bovine herpesvirus 1 (BHV-1)-seropositive animals would result in the bulk milk of a clean herd becoming BHV-1-positive. Probability calculations (stochastic and deterministic) were based on the distribution of the log(titre) of 828 positive animals and the daily milk production of the herds and of the individual cows. They showed that the probability in average sized herds of 45 dairy cows is only between 10 and 25 per cent and that even in small herds of 25 cows the introduction of a positive animal would go undetected in the majority of cases. It is concluded that if the bulk milk has become BHV-1-positive it is most likely that the infection has spread.

  12. Electrical field assisted matrix solid phase dispersion as a powerful tool to improve the extraction efficiency and clean-up of fluoroquinolones in bovine milk.

    Science.gov (United States)

    da Silva, Mariana Cristina; Orlando, Ricardo Mathias; Faria, Adriana Ferreira

    2016-08-26

    This work presents a new method by electrical matrix solid phase dispersion for the extraction and clean-up of marbofloxacin, ofloxacin, norfloxacin, ciprofloxacin, enrofloxacin, difloxacin and sarafloxacin in bovine milk. Composition and pH of the eluent, applied electrical potential and polarity were optimized by experimental designs. The combination of the chromatographic and electrophoretic mechanisms allowed the extraction and clean-up in one step with low organic solvent consumption, high extraction throughput and elution automation. Linearity, precision, trueness and limit of quantification were evaluated and provided values in accordance with other methods recently developed for the analysis of fluoroquinolones in milk. This technique proved to be promising for the extraction and clean-up of ionizable analytes in different milk matrices.

  13. Challenges for bovine viral diarrhoea virus antibody detection in bulk milk by antibody enzyme-linked immunosorbent assays due to changes in milk production levels

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Enøe, Claes; Stockmarr, Anders

    2015-01-01

    successfully been used to eradicate BVD in Sweden. Data (2003–2010) on changes in median herd size and milk production levels, occurrence of viremic animals and bulk milk surveillance were analysed. Additionally, the Danish blocking ELISA and the SVANOVIR ELISA were compared analyzing milk and serum samples....... The prevalence of antibody positive milking cows that could be detected by each test was estimated, by diluting positive individual milk samples and making artificial milk pools. Results: During the study period, the median herd size increased from 74 (2003) to 127 cows (2010), while the prevalence of BVDV...... infected herds decreased from 0.51 to 0.02 %. The daily milk yield contribution of a single seropositive cow to the entire daily bulk milk was reduced from 1.61 % in 2003 to 0.95 % in 2010 due to the increased herd size. It was observed that antibody levels in bulk milk decreased at national level...

  14. Evaluation of a chemiluminescence method for measuring alkaline phosphatase activity in whole milk of multiple species and bovine dairy drinks: interlaboratory study.

    Science.gov (United States)

    Salter, Robert S; Fitchen, John

    2006-01-01

    Alkaline phosphatase (ALP) is a ubiquitous enzyme in milk with time-temperature destruction similar to that of certain pathogens destroyed in pasteurization. Measurement of ALP to indicate proper pasteurization is a common practice. Recently the public health level for ALP was decreased to 350 mU/L, a level below the sensitivity of older colorimetric ALP methods. This study was conducted within the structure of the International Dairy Federation and the International Organization for Standardization to evaluate the reproducibility of the chemiluminescence method (Charm PasLite) for ALP at 50, 100, 350, and 500 mU/L in whole milk of multiple species to meet new regulations in the United States and proposed regulations in the European Union (EU). Fifteen laboratories from 8 countries evaluated bovine, goat, sheep, and buffalo milk, bovine skim milk, 20% fat cream, and 2% fat chocolate milk. At ALP levels of 350 and 500 mU/L, the average relative standard deviation for repeatability (RSDr) was 7.5%, and the average relative standard deviation of reproducibility was (RSDR) 15%. For ALP at 100 and 50 mU/L, the average RSDr values were 10.5 and 12.6%, respectively, and the average RSDR values were 18 and 25%, respectively. The limit of detection was 20 mU/L. Results are comparable to those obtained with other enzymatic photo-activated system methods such as the fluorometric method. Results indicate that the method is suitable for measuring ALP in the milk of multiple species and in dairy drinks at U.S. and proposed EU levels.

  15. Assessment of an extraction protocol to detect the major mastitis-causing pathogens in bovine milk.

    Science.gov (United States)

    Cressier, B; Bissonnette, N

    2011-05-01

    Despite all efforts to control its spread, mastitis remains the most costly disease for dairy farmers worldwide. One key component of better control of this disease is identification of the causative bacterial agent during udder infections in cows. Mastitis is complex, however, given the diversity of pathogens that must be identified. Development of a rapid and efficient bacterial species identification tool is thus necessary. This study was conducted to demonstrate the feasibility of bacterial DNA extraction for the automated molecular detection of major mastitis-causing pathogens directly in milk samples to complement traditional microbiological identification. Extraction and detection procedures were designed and optimized to achieve detection in a respectable time frame, at a reasonable cost, and with a high throughput capacity. The following species were identified: Staphylococcus aureus, Escherichia coli, Streptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae, and Klebsiella spp. (including Klebsiella oxytoca and Klebsiella pneumoniae). The detection procedure includes specific genomic DNA amplification by multiplex PCR for each species, separation by capillary electrophoresis, and laser-assisted automated detection. The specificity of the primers was assessed with a panel of bacteria representing mastitis-negative control species. The extraction protocol comprised multiple steps, starting with centrifugation for fat removal, followed by heating in the presence of a cation exchange resin to trap divalent ions. The analytical sensitivity was 100 cfu/mL for milk samples spiked with Staph. aureus, Strep. dysgalactiae, and E. coli, with a tendency for K. pneumoniae. The detection limit was 500 cfu/mL for Strep. uberis and Strep. agalactiae. The overall diagnostic sensitivity (95.4%) and specificity (97.3%) were determined in a double-blind randomized assay by processing 172 clinical milk samples with microbiological characterization as the

  16. Influences of different thermal processings in milk, bovine meat and frog protein structure.

    Science.gov (United States)

    Coura Oliveira, Tatiana; Lopes Lima, Samuel; Bressan, Josefina

    2013-01-01

    Several studies have associated the digestibility of proteins to its imunogenic potential. Though, it was objectified to evaluate the impact of the thermal processing with high and low temperatures on the proteins structure of three types of foods, by means of the digestibility in vitro and electroforesis en gel de poliacrilamida. The pasteurize was observed in such a way, firing 95 ºC during 15 minutes, how much freeze dried causes qualitative and quantitative modifications of constituent proteins of the food. The most sensible proteins to the increasing thermal processing order were beef, frog meat, and the last, cow milk.

  17. Functional properties of butter oil made from bovine milk with experimentally altered fat composition.

    Science.gov (United States)

    Ortiz-Gonzalez, G; Jimenez-Flores, R; Bremmer, D R; Clark, J H; DePeters, E J; Schmidt, S J; Drackley, J K

    2007-11-01

    Modification of milk fat composition might be desirable to alter manufacturing characteristics or produce low saturated fat dairy products that more closely meet consumer dietary preferences. The aim of this research was to evaluate functional properties of butter oil obtained from milks with fat composition modified by altering the profile of long-chain fatty acids (FA) absorbed from the small intestine of cows. A control and 5 mixtures of long-chain free FA were infused into the abomasum of lactating dairy cows in a 6 x 6 Latin square design with 21-d periods. Treatments were 1) control (no FA infused), 2) mostly saturated FA (C16:C18 = 0.72), 3) low-linoleic palm FA (C16:C18 = 0.85), 4) palm FA (C16:C18 = 0.72), 5) soy FA (C16:C18 = 0.10), and 6) high-palmitic soy FA (C16:C18 = 0.68). All treatments included meat solubles and Tween 80 as emulsifiers. Solid fat content (from 0 to 40 degrees C), melting point, and force at fracture were determined in butter oil. Milk fat from cows infused with palm FA (treatment 4) exhibited functionality equal to or better than control butter oil. Infusion with palm FA increased amounts of triglyceride (TG) fractions with 48, 52, and 54 carbon numbers but decreased TG with 32, 34, 36, and 42 carbon numbers. Infusion with soy FA increased TG with 26, 38, 40, 52, and 54 carbon numbers but decreased TG with 34, 42, and 46 carbons. Infusion of the mostly saturated FA increased TG with 38, 50, 52, and 54 carbon numbers but decreased TG with 32, 34, and 42 carbon numbers. These TG groups were consistently correlated with functional properties of butter oils from different treatments. The content of palmitic acid is important for maintaining functionality in the presence of increased polyunsaturated FA. The composition of milk fat may be able to be optimized through nutritional manipulation of diets for dairy cows if the optimal composition of FA and TG is defined for a particular dairy product.

  18. Effects of a Bovine Lactoferrin Formulation from Cow’s Milk on Menstrual Distress in Volunteers: A Randomized, Crossover Study

    Directory of Open Access Journals (Sweden)

    Hiroshi M. Ueno

    2016-05-01

    Full Text Available Dysmenorrhea is a highly prevalent complaint and highly undiagnosed gynecologic condition. Dairy products have a potential in the management of menstrual distress, and bovine lactoferrin can help the subjective dysphoria associated with dysmenorrhea. In the present study, we aimed to investigate the effects of a lactoferrin formulation isolated from cow’s milk on menstrual symptoms in volunteers. A double-blind, randomized, placebo-controlled, crossover study of the iron-lactoferrin complex (FeLf was performed in thirty-five healthy Japanese women. Participants received the 150 mg FeLf (per day or placebo from day ten of the luteal phase to day four of the follicular phase. The Moos Menstrual Distress Questionnaire (MDQ was measured for menstrual distress, and heart rate variability was measured as an index of autonomic nerve balance during menses. A visual analog scale for menstrual pain, and a verbal rating scale for quality of life during the first three days of menstruation were measured. The MDQ score for the automatic nervous system subscale was lower and the parasympathetic nervous system activity was greater in FeLf than in placebo for intention-to-treat or per-protocol populations. The other variables were not different between the groups. No treatment-related side effects were observed during the study. The results indicate that FeLf can provide a beneficial effect on the psychological symptoms in women affected by menstrual distress.

  19. Shiga Toxin-Producing Escherichia coli Isolated from Bovine Mastitic Milk: Serogroups, Virulence Factors, and Antibiotic Resistance Properties

    Science.gov (United States)

    Momtaz, Hassan; Safarpoor Dehkordi, Farhad; Taktaz, Taghi; Rezvani, Amir; Yarali, Sajad

    2012-01-01

    The aim of this study was to detect the virulence factors, serogroups, and antibiotic resistance properties of Shiga toxin-producing Escherichia coli, by using 268 bovine mastitic milk samples which were diagnosed using California Mastitis Test. After E. coli identification, PCR assays were developed for detection of different virulence genes, serogroups, and antibiotic resistance genes of Escherichia coli. The antibiotic resistance pattern was studied using disk diffusion method. Out of 268 samples, 73 (27.23%) were positive for Escherichia coli, and, out of 73 positive samples, 15 (20.54%) were O26 and 11 (15.06%) were O157 so they were the highest while O111 was not detected in any sample so it was the lowest serogroup. Out of 73 STEC strains, 11 (15.06%) and 36 (49.31%) were EHEC and AEEC, respectively. All of the EHEC strains had stx1, eaeA, and ehly, virulence genes, while in AEEC strains stx1 had the highest prevalence (77.77%), followed by eaeA (55.55%). Totally, aadA1 (65.95%) had the highest while blaSHV (6.38%) had the lowest prevalence of antibiotic resistance genes. The disk diffusion method showed that the STEC strains had the highest resistance to penicillin (100%), followed by tetracycline (57.44%), while resistance to cephalothin (6.38%) was the lowest. PMID:23213293

  20. Concentrated bovine milk whey active proteins facilitate osteogenesis through activation of the JNK-ATF4 pathway.

    Science.gov (United States)

    Tsuji-Naito, Kentaro; Jack, Ralph W

    2012-01-01

    Concentrated fractions of low molecular weight whey proteins (1-30 kDa), that is concentrated bovine milk whey active proteins (CBP), have been found to enhance bone formation in both in vivo and clinical studies, but the underlying mechanisms are poorly understood. In this study, we found that CBP promoted osteoblastic differentiation in normal human osteoblasts, and determined the involvement of the c-jun NH2-terminal kinase (JNK)-activating transcription factor 4 (ATF4) pathway. We observed that alkaline phosphatase activity and mineralization were significantly induced by CBP treatment. In addition, mRNA expression of ATF4 was intensely elevated in CBP-treated osteoblasts, indicating that the late-phase events of differentiation were promoted. We found that CBP activated the phosphorylation of JNK and extracellular signal-regulated kinase (ERK). Furthermore, pathway analyses using the various signaling pathway-specific inhibitors revealed that JNK activation, but not ERK activation, is essential for CBP-induced mineralization and ATF4 expression. Our results indicate that the JNK-mediated ATF4 pathway is required for CBP-promotive osteogenesis.

  1. HPLC Analysis of α-lactalbumin and β-lactoglobulin in Bovine Milk with C4 and C18 Column

    Institute of Scientific and Technical Information of China (English)

    Kong Xiao-yu; Wang Jing; Tang Yan-jun; Li Dan-dan; Zhang Nan-nan; Jiang Jin-dou; Liu Ning

    2012-01-01

    Reversed-phase high-performance liquid chromatography (RP-HPLC) method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column (250 mm×4.6 mm×5μm) and Shiseido CAPCELL PAK SG 300 C18 column (250 mm× 4.6 mm×5 μm) were used in the experiment. Phase A was composed of 0.1% (V/V) trifluoroacetic acid (TFA) in ultrapure water and Phase B (organic phase) was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.

  2. Effectiveness of convective drying to conserve indigenous yeasts with high volatile profile isolated from algerian fermented raw bovine milk (Rayeb

    Directory of Open Access Journals (Sweden)

    Latifa HAMOUDI-BELARBI

    2016-01-01

    Full Text Available Abstract Yeasts Candida tropicalis, Yarrowia lipolytica, Wickerhamomyces anomalus, Issatchenkia orientalis, Kluyveromyces marxianus, Saprochaete suaveolens and Trichosporon coremiiforme were isolated and identified by physiological, biochemical tests with API 20C AUX system and molecular methods by restriction fragment analysis of PCR-amplified 28S-rRNA from Algerian fermented raw bovine milk (Rayeb. Selected yeasts S. suaveolens, I. orientalis, K. marxianus and W. anomalus produced esters and higher esters which can exert a pertinent influence on the sensory characteristics of Rayeb. Viability of S. suaveolens and W. anomalus using three methods of drying (freeze-drying, convective drying, and spray-drying and during 4 months of storage at 4 °C and 25 °C in the darkness was studied. Immediately after each drying method, high survival was obtained using freeze-drying followed by convective drying in rice cakes and spray-drying respectively. During storage at 4 °C, convective drying provided better survival of yeast cultures of S. suaveolens and W. anomalus than freeze-drying. At 25 °C of storage, convective and freeze-dried yeast cultures showed no significant loss of viable cells up to 2 months of storage. Spray-dried yeast cultures had the greatest loss of viable count during the 3 months of storage at 25 °C.

  3. Investigation of the migration of triclabendazole residues to milk products manufactured from bovine milk, and stability therein, following lactating cow treatment.

    Science.gov (United States)

    Power, C; Danaher, M; Sayers, R; O'Brien, B; Clancy, C; Furey, A; Jordan, K

    2013-10-01

    Triclabendazole (TCB) is a flukicide used in the treatment of liver fluke in cattle; however, its use is currently prohibited in lactating dairy cows. In this study, following administration of 10% Fasinex (triclabendazole, Novartis Animal Health UK Ltd., Camberley, UK) the milk of 6 animals was used to manufacture dairy products, to ascertain if TCB residues in milk migrate into dairy products. The detection limit of the ultra-high-performance liquid chromatography-tandem mass spectrometry method used was 0.67 μg/kg. The highest concentrations of TCB residue measured, within the individual cow milk yield, was 1,529 ± 244 µg/kg (n=6), on d 2 posttreatment. Days 2 and 23 posttreatment represented high and low residue concentrations, respectively. At each of these 2 time points, the milk was pooled into 2 independent aliquots and refrigerated. Milk products, including cheese, butter, and skim milk powder were manufactured using pasteurized and unpasteurized milk from each aliquot. The results for high residue milks demonstrated that TCB residues concentrated in the cheese by a factor of 5 (5,372 vs. 918 µg/kg for cheese vs. milk) compared with the starting milk. Residue concentrations are the sum of TCB and its metabolites, expressed as keto-TCB. Residues were concentrated in the butter by a factor of 9 (9,177 vs. 1,082 μg/kg for butter vs. milk) compared with the starting milk. For milk, which was separated to skim milk and cream fractions, the residues were concentrated in the cream. Once skim milk powder was manufactured from the skim milk fraction, the residue in powder was concentrated 15-fold compared with the starting skim milk (7,252 vs. 423 µg/kg for powder vs. skim milk), despite the high temperature (185 °C) required during powder manufacture. For products manufactured from milk with low residue concentrations at d 23 posttreatment, TCB residues were detected in butter, cheese, and skim milk powder, even though there was no detectable residue in the

  4. The bovine ATP-binding cassette transporter ABCG2 Tyr581Ser single-nucleotide polymorphism increases milk secretion of the fluoroquinolone danofloxacin.

    Science.gov (United States)

    Otero, Jon A; Real, Rebeca; de la Fuente, Álvaro; Prieto, Julio G; Marqués, Margarita; Álvarez, Ana I; Merino, Gracia

    2013-03-01

    The bovine adenosine triphosphate-binding cassette transporter G2 (ABCG2/breast cancer resistance protein) polymorphism Tyr581Ser (Y581S) has recently been shown to increase in vitro transepithelial transport of antibiotics. Since this transporter has been extensively related to the active secretion of drugs into milk, the potential in vivo effect of this polymorphism on secretion of xenobiotics in livestock could have striking consequences for milk production, the dairy industry, and public health. Our purpose was to study the in vivo effect of this polymorphism on the secretion of danofloxacin, a widely used veterinary antibiotic, into milk. Danofloxacin (1.25 mg/kg) was administered to six Y/Y 581 homozygous and six Y/S 581 heterozygous lactating cows, and plasma and milk samples were collected and analyzed by high-performance liquid chromatography. No differences were found in the pharmacokinetic parameters of danofloxacin in plasma between the two groups of animals. In contrast, Y/S heterozygous cows showed a 2-fold increase in danofloxacin levels in milk. In addition, the pharmacokinetic elimination parameters, mean residence time and elimination half-life, were significantly lower in the milk of the animals carrying the Y/S polymorphism. These in vivo results are in agreement with our previously published in vitro data, which showed a greater capacity of the S581 variant in accumulation assays, and demonstrate, for the first time, an important effect of the Y581S single-nucleotide polymorphism on antibiotic secretion into cow milk. These findings could be extended to other ABCG2 substrates, and may be relevant for the treatment of mastitis and for the design of accurate and novel strategies to handle milk residues.

  5. N2 Gas Flushing Alleviates the Loss of Bacterial Diversity and Inhibits Psychrotrophic Pseudomonas during the Cold Storage of Bovine Raw Milk.

    Science.gov (United States)

    Gschwendtner, Silvia; Alatossava, Tapani; Kublik, Susanne; Fuka, Mirna Mrkonjić; Schloter, Michael; Munsch-Alatossava, Patricia

    2016-01-01

    The quality and safety of raw milk still remains a worldwide challenge. Culture-dependent methods indicated that the continuous N2 gas-flushing of raw milk reduced the bacterial growth during cold storage by up to four orders of magnitude, compared to cold storage alone. This study investigated the influence of N2 gas-flushing on bacterial diversity in bovine raw-milk samples, that were either cold stored at 6°C or additionally flushed with pure N2 for up to one week. Next-generation sequencing (NGS) of the V1-V2 hypervariable regions of 16S rRNA genes, derived from amplified cDNA, which was obtained from RNA directly isolated from raw-milk samples, was performed. The reads, which were clustered into 2448 operational taxonomic units (OTUs), were phylogenetically classified. Our data revealed a drastic reduction in the diversity of OTUs in raw milk during cold storage at 6°C at 97% similarity level; but, the N2-flushing treatment alleviated this reduction and substantially limited the loss of bacterial diversity during the same cold-storage period. Compared to cold-stored milk, the initial raw-milk samples contained less Proteobacteria (mainly Pseudomonadaceae, Moraxellaceae and Enterobacteriaceae) but more Firmicutes (mainly Ruminococcaceaea, Lachnospiraceae and Oscillospiraceaea) and Bacteroidetes (mainly Bacteroidales). Significant differences between cold-stored and additionally N2-flushed milk were mainly related to higher levels of Pseudomononadaceae (including the genera Pseudomonas and Acinetobacter) in cold-stored milk samples; furthermore, rare taxa were better preserved by the N2 gas flushing compared to the cold storage alone. No major changes in bacterial composition with time were found regarding the distribution of the major 9 OTUs, that dominated the Pseudomonas genus in N2-flushed or non-flushed milk samples, other than an intriguing predominance of bacteria related to P. veronii. Overall, this study established that neither bacteria causing milk

  6. The occurrence of noncoagulating milk and the association of bovine milk coagulation properties with genetic variants of the caseins in 3 Scandinavian dairy breeds

    DEFF Research Database (Denmark)

    Poulsen, Nina Aagaard; Bertelsen, Henriette Pasgaard; Jensen, Hanne Bak

    2013-01-01

    Substantial variation in milk coagulation properties has been observed among dairy cows. Consequently, raw milk from individual cows and breeds exhibits distinct coagulation capacities that potentially affect the technological properties and milk processing into cheese. This variation is largely...... breeds, which more or less reflected the single gene results. Overall, our results strongly suggest that milk coagulation is under the influence of additive genetic variation. Optimal milk for future cheese production can be ensured by monitoring the frequency of unfavorable variants and thus preventing...... an increase in the number of cows producing milk with impaired coagulation. Selective breeding for variants associated with superior milk coagulation can potentially increase raw milk quality and cheese yield in all 3 Scandinavian breeds...

  7. Diurnal differences in milk composition and its influence on in vitro growth of Staphylococcus aureus and Escherichia coli in bovine quarter milk

    NARCIS (Netherlands)

    Eisenberg, S.W.F.; Boerhout, E.M.; Ravesloot, L.; Daemen, A.J.J.M.; Benedictus, L.; Rutten, V.P.M.G.; Koets, A.P.

    2016-01-01

    In experimental intramammary inoculation studies, it has been observed that mastitis susceptibility is influenced, among others, by cow factors. To identify milk characteristics leading to these differences, quarter milk samples of morning and evening milk were collected and analyzed for their co

  8. Effect of Supplementation with Antioxidants on the Quality of Bovine Milk and Meat Production

    Directory of Open Access Journals (Sweden)

    Cristina Castillo

    2013-01-01

    Full Text Available From a clinical point of view, oxidative stress (OS is considered the primary cause of numerous metabolic processes in transition cow. Thus, the addition of antioxidants has been considered a palliative or preventive treatment. But beyond the clinical perspective, antioxidant supplementation provides an added value to the product obtained being either milk or meat. This paper reviews the beneficial aspects that provide antioxidant supplementation on quality of both products and that fit into the new concept that the consumer has a functional and healthy food. Our approach is from a veterinary standpoint, by reviewing the studies conducted to date and the new perspectives that are interesting and need to be studied in the following years. One of the highlights is that sustainable farming, one in which production is combined with animal health, also impacts positively on the quality of the final products, with beneficial antioxidant properties to human health.

  9. Effect of supplementation with antioxidants on the quality of bovine milk and meat production.

    Science.gov (United States)

    Castillo, Cristina; Pereira, Víctor; Abuelo, Ángel; Hernández, Joaquín

    2013-01-01

    From a clinical point of view, oxidative stress (OS) is considered the primary cause of numerous metabolic processes in transition cow. Thus, the addition of antioxidants has been considered a palliative or preventive treatment. But beyond the clinical perspective, antioxidant supplementation provides an added value to the product obtained being either milk or meat. This paper reviews the beneficial aspects that provide antioxidant supplementation on quality of both products and that fit into the new concept that the consumer has a functional and healthy food. Our approach is from a veterinary standpoint, by reviewing the studies conducted to date and the new perspectives that are interesting and need to be studied in the following years. One of the highlights is that sustainable farming, one in which production is combined with animal health, also impacts positively on the quality of the final products, with beneficial antioxidant properties to human health.

  10. Effect of insoluble calcium concentration on endogenous syneresis rate in rennet-coagulated bovine milk.

    Science.gov (United States)

    Choi, J; Horne, D S; Lucey, J A

    2015-09-01

    The rennet coagulation of milk has been extensively studied. Mathematical modeling of the gelation process has been performed, mainly for the purpose of predicting the gel point. Rheological profiles of rennet gels during aging (long reaction times) have indicated that the gel stiffness (modulus) attains a maximum and thereafter decreases. We wanted to model this type of behavior and used the Carlson model, which includes terms for the proteolysis of κ-casein hairs (creating active sites) and the crosslinking of these activated sites. To account for the observed decrease in the gel modulus with time, we modified the Carlson model by adding an exponential decay term, which we ascribe to endogenous syneresis. We believe that this decay (i.e., syneresis rate) would likely be influenced by the mobility of bonds within casein micelles (in gels as indicated by the rheological loss tangent parameter). To modify the internal structural bonding of casein micelles, reconstituted skim milk was acidified to pH values 6.4, 6.0, 5.8, 5.6, and 5.4, or EDTA was added to milk at concentrations of 0, 2, 4, and 6mM, and the final pH values of EDTA-treated samples were subsequently adjusted to pH 6.0. These treatments were then used to prepare rennet gel samples that were monitored by dynamic low amplitude oscillatory rheometry. When the modified Carlson model was fitted to the actual experimental storage modulus values of each sample, it fitted the data reasonably well (especially the pH trial data). As the pH values of milk decreased, the modulus values at infinite reaction time (G'∞) increased; however, G'∞ decreased with an increase in the EDTA concentration. In the pH trial, the rate constants for the proteolysis of κ-casein hairs and the crosslinking of these activated sites exhibited a maximum at pH 5.6 and 6.0, respectively. The rate constant for endogenous syneresis increased at pH values syneresis was significantly positively correlated (r≥0.96) with the loss tangent

  11. Screening of bovine milk samples for sub-clinical mastitis and antibiogram of bacterial isolates

    Directory of Open Access Journals (Sweden)

    Harini H. and Sumathi B.R.

    Full Text Available The study was undertaken to find out the incidence of subclinical mastitis (SCM and to assess the antibiotic sensitivity pattern of the causative organisms in lactating cows in and around Kanakapura taluk, Ramanagara district of Karnataka state. The prevalence of subclinical mastitis was assessed by the results of 3 different screening tests and bacteriological evaluation was done for the milk samples that were found positive. The predominant bacterial isolates recovered were Staphylococcus aureus (58% and Escherichia coli (23.5% followed by Staphylococcus epidermidis (8%, Streptococcus sp. (5.5%, Klebsiella sp. (3% and Bacillus sp. (2%. The in vitro antibiogram studies of bacterial isolates revealed higher sensitivity for ciprofloxacin (89%, ofloxacin (85%, enrofloxacin (82%, gentamicin (80% and chloramphenicol (75%, resistant to colistin, neomycin, streptomycin, penicillin and tetracycline. [Vet. World 2011; 4(8.000: 358-359

  12. Xanthine oxidase biosensor for monitoring meat spoilage

    Science.gov (United States)

    Vanegas, D. C.; Gomes, C.; McLamore, E. S.

    2014-05-01

    In this study, we have designed an electrochemical biosensor for real-time detection of specific biomarkers of bacterial metabolism related to meat spoilage (hypoxanthine and xanthine). The selective biosensor was developed by assembling a `sandwich' of nanomaterials and enzymes on a platinum-iridium electrode (1.6 mm tip diameter). The materials deposited on the sensor tip include amorphous platinum nanoclusters (i.e. Pt black), reduced graphene oxide, nanoceria, and xanthine oxidase. Xanthine oxidase was encapsulated in laponite hydrogel and used for the biorecognition of hypoxanthine and xanthine (two molecules involved in the rotting of meat by spoilage microorganisms). The developed biosensor demonstrated good electrochemical performance toward xanthine with sensitivity of 2.14 +/- 1.48 μA/mM, response time of 5.2 +/- 1.5 sec, lower detection limit of 150 +/- 39 nM, and retained at least 88% of its activity after 7 days of continuous use.

  13. The comparative studies of proteins of bovine and goat milk%牛羊乳蛋白组分比较研究

    Institute of Scientific and Technical Information of China (English)

    宋宏新; 张小苗; 薛海燕

    2012-01-01

    Goat milk, which hailed as "The king of milk" by international nutrition academic, has been gradually accepted as daily life's nutritional supplements by people. This paper reviewed the differences of composition of protein (mainly the caseins and whey proteins), composition of amino acids and variants between proteins of goat milk and bovine milk, and the differences of casein micelle, as well, which provides a theoretical basis for the testing and processing of goat milk.%羊乳被国际营养学界誉为“奶中之王”,逐渐被人们列为日常生活的营养保健佳品.该文对羊乳和牛乳的蛋白组分(主要是酪蛋白和乳清蛋白)含量、氨基酸组成及变异体等方面的差异进行了综述,并且对两者酪蛋白胶束的差异进行了比较,为羊乳检验和加工提供理论依据.

  14. Development of an LC-MS/MS method to quantify sex hormones in bovine milk and influence of pregnancy in their levels.

    Science.gov (United States)

    Regal, P; Cepeda, A; Fente, C

    2012-01-01

    Hormones work in harmony in the body, and this status must be maintained to avoid metabolic disequilibrium and the subsequent illness. Besides, it has been reported that exogenous steroids (presence in the environment and food products) influence the development of several important illnesses in humans. Endogenous steroid hormones in food of animal origin are unavoidable as they occur naturally in these products. The presence of hormones in food has been connected with several human health problems. Bovine milk contains considerable quantities of hormones and it is of particular concern. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, based on hydroxylamine derivatisation, has been developed and validated for the quantification of six sex hormones in milk [pregnenolone (P₅), progesterone (P₄), estrone (E₁), testosterone (T), androstenedione (A) and dehydroepiandrosterone (DHEA)]. This method has been applied to real raw milk samples and the existence of differences between milk from pregnant and non-pregnant cows has been statistically confirmed. Basing on a revision of existing published data, it could be concluded that maximum daily intakes for hormones are not reached through milk ingestion. Although dairy products are an important source of hormones, other products of animal origin must be considered as well for intake calculations.

  15. Diurnal differences in milk composition and its influence on in vitro growth of Staphylococcus aureus and Escherichia coli in bovine quarter milk.

    Science.gov (United States)

    Eisenberg, S W F; Boerhout, E M; Ravesloot, L; Daemen, A J J M; Benedictus, L; Rutten, V P M G; Koets, A P

    2016-07-01

    In experimental intramammary inoculation studies, it has been observed that mastitis susceptibility is influenced, among others, by cow factors. To identify milk characteristics leading to these differences, quarter milk samples of morning and evening milk were collected and analyzed for their composition (protein, fat, lactose, urea, lactoferrin, lactoperoxidase, and β-lactoglobulin concentrations), somatic cell count, and antibodies against Staphylococcus aureus. Furthermore, in vitro growth of S. aureus and Escherichia coli in fresh quarter milk samples was determined. All measured parameters differed significantly between quarters and also between morning and evening milk with the exception of lactose levels. In addition, quantitative growth of S. aureus and E. coli was significantly different in morning milk compared with evening milk. Mixed model analysis revealed that replication of S. aureus was negatively associated with the presence of fat, S. aureus-specific IgG1 antibodies, contamination of the milk sample and morning milk. Replication of E. coli was negatively associated with fat concentrations, and positively associated with morning milk. The significant difference between morning and evening milk supports the theory that changes in milk composition influence bacterial growth. Although all determined milk components differed significantly between quarters and in time no significant association with bacterial growth could be identified with the exception of fat for both studied species and IgG1 titers for S. aureus. The negative association of fat with bacterial growth was assumed to occur due to activation of lipolysis by milk handling and can most likely be neglected for in vivo relevance. The fact that S. aureus-specific IgG1 titers were negatively associated with S. aureus growth in vitro encourages the ongoing effort to develop a vaccine against S. aureus-induced mastitis.

  16. Effects of cell culture techniques on gene expression and cholesterol efflux in primary bovine mammary epithelial cells derived from milk and tissue.

    Science.gov (United States)

    Sorg, D; Potzel, A; Beck, M; Meyer, H H D; Viturro, E; Kliem, H

    2012-10-01

    Primary bovine mammary epithelial cells (pbMEC) are often used in cell culture to study metabolic and inflammatory processes in the udder of dairy cows. The most common source is udder tissue from biopsy or after slaughter. However, it is also possible to culture them from milk, which is non-invasive, repeatable and yields less contamination with fibroblasts. Generally, not much is known about the influence of cell origin and cell culture techniques such as cryopreservation on pbMEC functionality. Cells were extracted from milk and udder tissue to evaluate if milk-derived pbMEC are a suitable alternative to tissue-derived pbMEC and to test what influence cryopreservation has. The cells were cultivated for three passages and stored in liquid nitrogen. The relative gene expression of the five target genes kappa-casein, lingual antimicrobial peptide (LAP), lactoferrin, lysozyme (LYZ1) and the prolactin receptor normalised with keratin 8 showed a tendency to decrease in the tissue cultures, but not in the milk-derived cultures, suggesting a greater influence of the cultivation process on tissue-derived cells, freezing lowered expression levels in both cultures. Overall expression of LAP and LYZ1 tended to be higher in milk cells. Cholesterol efflux was measured to compare passages one to seven in milk-derived cells. Passage number did not alter the efflux rate (p ≤ 0.05). We showed for the first time that the extraction of pbMEC from milk can be a suitable alternative to tissue extraction.

  17. Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells.

    Science.gov (United States)

    Xia, Xiaojing; Che, Yanyi; Gao, Yuanyuan; Zhao, Shuang; Ao, Changjin; Yang, Hongjian; Liu, Juxiong; Liu, Guowen; Han, Wenyu; Wang, Yuping; Lei, Liancheng

    2016-05-31

    During the lactation cycle of the bovine mammary gland, autophagy is induced in bovine mammary epithelial cells (BMECs) as a cellular homeostasis and survival mechanism. Interferon gamma (IFN-γ) is an important antiproliferative and apoptogenic factor that has been shown to induce autophagy in multiple cell lines in vitro. However, it remains unclear whether IFN-γ can induce autophagy and whether autophagy affects milk synthesis in BMECs. To understand whether IFN-γ affects milk synthesis, we isolated and purified primary BMECs and investigated the effect of IFN-γ on milk synthesis in primary BMECs in vitro. The results showed that IFN-γ significantly inhibits milk synthesis and that autophagy was clearly induced in primary BMECs in vitro within 24 h. Interestingly, autophagy was observed following IFN-γ treatment, and the inhibition of autophagy can improve milk protein and milk fat synthesis. Conversely, upregulation of autophagy decreased milk synthesis. Furthermore, mechanistic analysis confirmed that IFN-γ mediated autophagy by depleting arginine and inhibiting the general control nonderepressible-2 kinase (GCN2)/eukaryotic initiation factor 2α (eIF2α) signaling pathway in BMECs. Then, it was found that arginine supplementation could attenuate IFN-γ-induced autophagy and recover milk synthesis to some extent. These findings may not only provide a novel measure for preventing the IFN-γ-induced decrease in milk quality but also a useful therapeutic approach for IFN-γ-associated breast diseases in other animals and humans.

  18. Purification of MUC1 from bovine milk-fat globules and characterization of a corresponding full-length cDNA clone

    DEFF Research Database (Denmark)

    Pallesen, Lone Tjener; Andersen, Mikkel Holmen; Nielsen, Rune L.;

    2001-01-01

    The highly glycosylated protein MUC1 was purified from bovine milk-fat globule membranes by a procedure involving detergent extraction, ion-exchange chromatography and reverse-phase chromatography. The identity of the purified mucin protein was confirmed by N-terminal sequencing and partial amino...... amino acid sequence demonstrated structural features characteristic for mucins, including an extracellular tandem repeat region with 11 partially conserved repeats (20 amino acids each), a membrane-proximal SEA module, a transmembrane domain, and a cytoplasmic C-terminal region. Monosaccharide...

  19. Perspective on calf and mammary gland development through changes in the bovine milk proteome over a complete lactation

    NARCIS (Netherlands)

    Zhang, L.; Boeren, J.A.; Hageman, J.A.; Hooijdonk, van A.C.M.; Vervoort, J.J.M.; Hettinga, K.A.

    2015-01-01

    Milk contains all the nutrients for the growth and development of the neonate. However, milk composition is not constant during lactation. To study the changes of the milk proteome over lactation, filter-aided sample preparation combined with dimethyl labeling followed by liquid chromatography tande

  20. Deep Sequencing and Screening of Differentially Expressed MicroRNAs Related to Milk Fat Metabolism in Bovine Primary Mammary Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Binglei Shen

    2016-02-01

    Full Text Available Milk fat is a key factor affecting milk quality and is also a major trait targeted in dairy cow breeding. To determine how the synthesis and the metabolism of lipids in bovine milk is regulated at the miRNA level, primary mammary epithelial cells (pMEC derived from two Chinese Holstein dairy cows that produced extreme differences in milk fat percentage were cultured by the method of tissue nubbles culture. Small RNA libraries were constructed from each of the two pMEC groups, and Solexa sequencing and bioinformatics analysis were then used to determine the abundance of miRNAs and their differential expression pattern between pMECs. Target genes and functional prediction of differentially expressed miRNAs by Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes analysis illustrated their roles in milk fat metabolism. Results show that a total of 292 known miRNAs and 116 novel miRNAs were detected in both pMECs. Identification of known and novel miRNA candidates demonstrated the feasibility and sensitivity of sequencing at the cellular level. Additionally, 97 miRNAs were significantly differentially expressed between the pMECs. Finally, three miRNAs including bta-miR-33a, bta-miR-152 and bta-miR-224 whose predicted target genes were annotated to the pathway of lipid metabolism were screened and verified by real-time qPCR and Western-blotting experiments. This study is the first comparative profiling of the miRNA transcriptome in pMECs that produce different milk fat content.

  1. Bovine milk-based formula leads to early maturation-like morphological, immunological, and functional changes in the jejunum of neonatal piglets.

    Science.gov (United States)

    Pieper, R; Scharek-Tedin, L; Zetzsche, A; Röhe, I; Kröger, S; Vahjen, W; Zentek, J

    2016-03-01

    Artificial rearing and formula feeding is coming more into the focus due to increasing litter sizes and limited nursing capacity of sows. The formula composition is important to effectively support the development of the gut and prevent intestinal dysfunction in neonatal piglets. In this study, newborn piglets ( = 8 per group) were fed a bovine milk-based formula (FO), containing skimmed milk and whey as the sole protein and carbohydrate sources, or were suckled by the sow (sow milk [SM]). After 2 wk, tissue from the jejunum was analyzed for structural (i.e., morphometry) and functional (i.e., disaccharidase activity, glucose transport, permeability toward macromolecules, and immune cell presence) changes and concomitant expression of related genes. Formula-fed piglets had more liquid feces ( < 0.05) over the entire experimental period. Although FO contained twice as much lactose (46% on a DM basis) as SM (21%) and no maltose or starch, the lactase activity was lower ( < 0.05) and glucose transport capacity was higher ( < 0.05) in FO-fed pigs. The relative proportion of intraepithelial natural killer cells and proinflammatory cytokine gene expression (, , and ) was higher in FO-fed pigs ( < 0.05). Piglets fed FO had deeper crypts, larger villus area, and higher expression of caspase 3 and proliferating cell nuclear antigen ( < 0.05). Epithelial permeability toward fluorescein isothiocyanate-dextran was higher and expression of claudin-4 was lower in FO-fed piglets ( < 0.05). The data suggest an early response to bovine milk-based compounds in the FO accompanied with early onset of functional maturation and impaired barrier function. Whether lactose, absence of species-specific protective factors, or antigenicity of foreign proteins lead to to the observed intestinal reactions requires further clarification.

  2. Research of nitroxynil residues in bovine milk following a single administration in the dry period by ultra-performance liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Claudia Chirollo

    2013-10-01

    Full Text Available Nitroxynil (NIT is a halogenated phenol used to control fascioliasis in cattle and sheep. The Commission Regulation EU No 37/2010 has established maximum residue limits for NIT in bovine and ovine muscle (400 μg kg−1, fat (200 μg kg−1, liver (20 μg kg−1 and kidney (400 μg kg−1, and more recently in bovine and ovine milk (20 μg kg−1. Thirty-five pregnant dairy cows were treated in this study with nitroxynil (340 mg/mL solution for injection at the recommended dose of 10 mg/kg body weight at the start of the dry period, i.e. 53 to 74 days before the expected calving. Calving occurred between 43 days and 79 days after treatment. The concentrations of NIT in the milk were monitored for up to 120 days after calving. NIT residues were extracted using acetonitrile; magnesium sulfate and sodium chloride were added to induce liquid-liquid partitioning and purified by dispersive solid phase extraction for clean-up. NIT was detected by ultra high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS in negative ionization mode. The highest concentrations of this drug were found in two animals at the first milking, 48 and 53 day post treatment with levels of 362 and 657 μg kg–1, respectively. NIT residues were below the limit of detection of the method (0.24 μg/kg–1 between 67 and 106 day post-treatment. Following calving, residues rapidly depleted in animals and were non-detectable from 10 to 38 days post-calving. In particular, in all animals milk resulted compliant (<20 μg/kg−1 three days post partum.

  3. 中国北方人初乳、牛初乳、牛常乳、牛血中胰岛素样生长因子-1和神经生长因子含量的比较%Comparison of IGF-1 and NGF in human colostrum, bovine colostrum,bovine milk and bovine blood serum in northern China

    Institute of Scientific and Technical Information of China (English)

    王洋; 生庆海; 张玉梅; 赵艾; 贾梦; 薛勇; 王培玉

    2011-01-01

    Objective To quantify and compare the levels of insulin-like growth factor 1 ( IGF-1 ) and nerve growth factor (NGF) in human colostrum, bovine colostrum, bovine milk and bovine blood serum. Methods Radioimmunoassay was used in the quantification of IGF-1 and NGF. Data were analyzed by using SPSS 13. 0. Results The levels of IGF-1 in human colostrum, bovine colostrum, bovine milk and bovine blood serum were 26. 91, 38.40, 20. 14 and 37.35 μg/L,respectively. The levels of NGF in human colostrum, bovine colostrum, milk and bovine blood serum were 300. 47,69.82, 110. 37 and 9. 63 ng/L, respectively. The differences between the levels of IGF-1 in human colostrum and bovine colostrum were not significant, but the levels of IGF-1 in bovine colostrum were obviously higher than that in bovine milk (P <0. 05), and there was no significant difference between the levels of IGF-1 in bovine milk and bovine blood serum.The level of NGF in human colostrum was obviously higher than that in bovine colostrum ( P < 0.05 ). There was no significant difference between the levels of NGF in bovine colostrum and bovine milk. The level of NGF in bovine blood serum was obviously lower than that in bovine milk ( P < 0. 05 ). Conclusion The level of IGF-1 in human colostrum was not significantly different from that in bovine colostrum, but the level of NGF in human colostrum was much higher than that in bovine colostrum.%目的 检测人初乳、集中饲养的新西兰进口荷斯坦乳牛的初乳、常乳和血液中胰岛素样生长因子-1(IGF-1)和神经生长因子(NGF)的含量,比较人初乳和牛初乳、牛初乳和牛常乳以及牛乳和同期采集的血液中IGF-1、NGF的含量.方法 采用放射免疫试剂盒测定人初乳、牛初乳、牛常乳和血中IGF-1、NGF的含量,使用SPSS 13.0进行统计分析.结果 本研究中测定的人初乳中IGF-1的含量是26.91μg/L,荷斯坦乳牛的初乳、常乳和血清中IGF-1的含量分别是38.40、20.14和37.35

  4. Research on Determination Method of Flunixin Residue in Bovine Milk and Milk Powder by HPLC-UV%牛奶和奶粉中氟胺烟酸残留量测定方法的研究

    Institute of Scientific and Technical Information of China (English)

    张晨; 苏海滨; 陶迎春; 赵静; 吴鹏飞; 杨晓旎; 蒋玉新; 宋文斌

    2013-01-01

    The method for the determination of flunixin residues in aquatic products is studied by HPLC-UV.Bovine milk is digested using β-glucuronidase.The enzyme digestion was extracted using acetonitrile,cleaned up with SPE column.The solution was analyzed by LC-UV using external standard method.The detection limit of this method is 5μg/kg for bovine milk and 40mg/kg for milk power.The results showed a good linear (r=0.9996) in the concentration range of 2.5-160.0μg/kg,the average recoveries for spiked standards (5.0μg/kg-40μg/kg)and(40.0μg/kg-100.0μg/kg) were 86.86%-100.32%,the relative standard deviations were 1.986%-2.295% in bovine milk and milk powder.%研究了牛奶和奶粉中氟胺烟酸残留量的液相色谱-紫外分析方法.样品在酸性条件下酶解用乙腈提取后,经固相革取柱净化,用配有紫外检测器的高效液相色谱测定,外标法定量.方法的最低检出限牛奶为5mg/kg,奶粉为40mg/kg;在2.5~160.0mg/kg浓度范围内,线性关系良好,相关系数r=0.9996,添加水平牛奶5.0~ 40mg/kg时,奶粉40.0~100mg/kg时,回收率在86.86%~100.32%,相对标准偏差(RSD)为1.986%~2.295%.

  5. Determination of polychlorinated biphenyls in ocean water and bovine milk using crosslinked polymeric ionic liquid sorbent coatings by solid-phase microextraction.

    Science.gov (United States)

    Joshi, Manishkumar D; Ho, Tien D; Cole, William T S; Anderson, Jared L

    2014-01-01

    Crosslinked polymeric ionic liquid (PIL)-based sorbent coatings were employed in the extraction of 21 polychlorinated biphenyls (PCBs) from ocean water and bovine milk using solid-phase microextraction (SPME). The extraction temperature, time, and concentration of sodium chloride added to the matrix were optimized in order to determine the best extraction conditions for the extraction of PCBs. The analytical performance of the crosslinked PIL-based SPME fibers was compared with a commercial 7 µm polydimethylsiloxane (PDMS) fiber using gas chromatography (GC) employing an electron capture detector (ECD) and mass spectrometric detection (MS). Higher sensitivities for PCBs were achieved using PIL-based fibers when compared to PDMS fiber due to the incorporation of benzyl moieties into the PIL structures. The limits of detection (LOD) for all PCBs were determined to be in the low ng L(-1) range using the three studied coatings. Recovery studies were performed for PCBs in ocean water and bovine milk to validate the applicability of the current SPME method.

  6. Quantitative determination of corticosteroids in bovine milk using mixed-mode polymeric strong cation exchange solid-phase extraction and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Tölgyesi, Adám; Tölgyesi, László; Sharma, Virender K; Sohn, Mary; Fekete, Jeno

    2010-12-01

    A new method was developed to identify and quantify corticosteroids (prednisolone, methylprednisone, flumetasone, dexamethasone, and methylprednisolone) in raw bovine milk by liquid chromatography-tandem mass spectrometry (LC-MS/MS) utilizing mixed-mode polymeric strong cation exchange and reversed-phase (MCX) solid-phase extraction (SPE) to reduce ion effects in a multimode ion (MMI) source. The main advantage of this method over other commonly used methods includes the use of a single SPE cartridge with a low volume for sample preparation and fast separation on the HPLC system with reduced ion suppression. This study is the first to report the determination of methylprednisone, a metabolite of methylprednisolone, in bovine milk. This method was validated in accordance with the European Union (EU) Commission Decision 2002/657/EC. The recoveries vary between 90% and 105%. The within-laboratory reproducibility (precision) is less than 30%. The decision limits and detection capabilities were calculated along with LODs, which ranged from 0.02 to 0.07 microg/kg. The method was further enhanced by its successful adaptation to other LC-MS/MS systems equipped with the newly developed ion source, Agilent Jet Stream (AJS). After optimization of the AJS ion source and MS parameters, even lower LOD values were achieved (0.001-0.006 microg/kg) for the corticosteroids. Analytical results obtained with the AJS were characterized by an enhanced area response and similar noise level comparable to those obtained with conventional orthogonal atmospheric ionization (API).

  7. Transglutaminase inhibitor from milk

    NARCIS (Netherlands)

    Jong, G.A.H. de; Wijngaards, G.; Koppelman, S.J.

    2003-01-01

    Cross-linking experiments of skimmed bovine milk with bacterial transglutaminase isolated from Streptoverticillium mobaraense showed only some degree of formation of high-molecular-weight casein polymers. Studies on the nature of this phenomenon revealed that bovine milk contains an inhibitor of tra

  8. Antimicrobial Susceptibility Patterns of Environmental Streptococci Recovered from Bovine Milk Samples in the Maritime Provinces of Canada

    Science.gov (United States)

    Cameron, Marguerite; Saab, Matthew; Heider, Luke; McClure, J Trenton; Rodriguez-Lecompte, Juan Carlos; Sanchez, Javier

    2016-01-01

    Determination of antimicrobial susceptibility of bovine mastitis pathogens is important for guiding antimicrobial treatment decisions and for the detection of emerging resistance. Environmental streptococci are ubiquitous in the farm environment and are a frequent cause of mastitis in dairy cows. The aim of the study was to determine patterns of antimicrobial susceptibility among species of environmental streptococci isolated from dairy cows in the Maritime Provinces of Canada. The collection consisted of 192 isolates identified in milk samples collected from 177 cows originating from 18 dairy herds. Results were aggregated into: (1) Streptococcus uberis (n = 70), (2) Streptococcus dysgalactiae (n = 28), (3) other Streptococci spp. (n = 35), (4), Lactococcus spp. (n = 32), and (5) Enterococcus spp. (n = 27). Minimum inhibitory concentrations (MICs) were determined using the Sensititre microdilution system and mastitis plate format. Multilevel logistic regression models were used to analyze the data, with antimicrobial susceptibility as the outcome. The proportion of susceptible S. uberis ranged from 23% (for penicillin) to 99% (for penicillin/novobiocin), with a median of 82%. All S. dysgalactiae were susceptible to all antimicrobials except for penicillin (93% susceptible) and tetracycline (18% susceptible). The range of susceptibility for other Streptococcus spp. was 43% (for tetracycline) to 100%, with a median percent susceptibility of 92%. Lactococcus spp. isolates displayed percent susceptibilities ranging from 0% (for penicillin) to 97% (for erythromycin), median 75%. For the antimicrobials tested, the minimum inhibitory concentrations were higher for Enterococcus spp. than for the other species. According to the multilevel models, there was a significant interaction between antimicrobial and bacterial species, indicating that susceptibility against a particular antimicrobial varied among the species of environmental streptococci and vice

  9. Effects of glucose availability on expression of the key genes involved in synthesis of milk fat, lactose and glucose metabolism in bovine mammary epithelial cells.

    Directory of Open Access Journals (Sweden)

    Hongyun Liu

    Full Text Available As the main precursor for lactose synthesis, large amounts of glucose are required by lactating dairy cows. Milk yield greatly depends on mammary lactose synthesis due to its osmoregulatory property for mammary uptake of water. Thus, glucose availability to the mammary gland could be a potential regulator of milk production. In the present study, the effect of glucose availability on expression of the key genes involved in synthesis of milk fat, lactose and glucose metabolism in vitro was investigated. Bovine mammary epithelial cells (BMEC were treated for 12 h with various concentrations of glucose (2.5, 5, 10 or 20 mmol/L. The higher concentrations of glucose (10-20 mmol/L did not affect the mRNA expression of acetyl-CoA carboxylase, diacyl glycerol acyl transferase, glycerol-3 phosphate acyl transferase and α-lactalbumin, whereas fatty acid synthase, sterol regulatory element binding protein-1 and beta-1, 4-galactosyl transferase mRNA expression increased at 10 mmol/L and then decreased at 20 mmol/L. The content of lactose synthase increased with increasing concentration of glucose, with addition of highest value at 20 mmol/L of glucose. Moreover, the increased glucose concentration stimulated the activities of pyruvate kinase and glucose-6-phosphate dehydrogenase, and elevated the energy status of the BMEC. Therefore, it was deduced that after increasing glucose availability, the extra absorbed glucose was partitioned to entering the synthesis of milk fat and lactose by the regulation of the mRNA expression of key genes, promoting glucose metabolism by glycolysis and pentose phosphate pathway as well as energy status. These results indicated that the sufficient availability of glucose in BMEC may promote glucose metabolism, and affect the synthesis of milk composition.

  10. Pérdidas económicas ocasionadas por la mastitis bovina en la industria lechera - Economic causalties inflicted by the bovine mastitis in the milk industry

    Directory of Open Access Journals (Sweden)

    Bedolla, CC;

    2008-04-01

    Full Text Available ResumenEl objetivo del presente trabajo fue hacer una revisión bibliográfica sobre las pérdidas económicas que ocasiona la mastitis bovina en la industria lechera en el Mundo, en México y Michoacán. La mastitis es considerada la enfermedad infecciosa más costosa de las vacas lecheras debido a que induce a una disminución en la producción del 4 al 30% de leche y baja su calidad, además de incrementar los costos del cuidado de la salud del hato y un desecho prematuro de animales genéticamente mejorados. En el documento, se hace una descripción de los conceptos concernientes a la mastitis bovina, su clasificación, características de la enfermedad tanto en su forma de presentación clínica y subclínica en el ganado bovino lechero, así mismo se hace referencia a las pérdidas económicas que ésta ocasiona tanto en la industria lechera a nivel mundial, como en México y en el Estado deMichoacán; finalmente, se aborda lo relacionado a los costos de prevención, tratamiento y control de la enfermedad. Se concluye que las pérdidas económicas que ocasiona la mastitis bovina a la industria lechera son considerables, esto en base a diversos estudios reportados por diferentes autores.summaryThe objective of the present paper was to make a revision of literature on the economic causalties that the bovine mastitis in the milk industry in the World inflicts, in Mexico and Michoacan. The mastitis is considered infectious the disease more expensive of the milk cows because it induces to a diminution in the production from the 4 to 30% of low milk and his quality, besides to increase the costs of the care of the health of the cattle ranch and a premature animal remainder genetically improved. In the document, a description of the concepts concerning the bovine mastitis, its classification,characteristics of the disease as much in its form of clinical and subclinical presentation in the milk bovine cattle becomes, also reference to the economic

  11. A New ELISA Using the ANANAS Technology Showing High Sensitivity to diagnose the Bovine Rhinotracheitis from Individual Sera to Pooled Milk.

    Science.gov (United States)

    Casarin, Elisabetta; Lucchese, Laura; Grazioli, Santina; Facchin, Sonia; Realdon, Nicola; Brocchi, Emiliana; Morpurgo, Margherita; Nardelli, Stefano

    2016-01-01

    Diagnostic tests for veterinary surveillance programs should be efficient, easy to use and, possibly, economical. In this context, classic Enzyme linked ImmunoSorbent Assay (ELISA) remains the most common analytical platform employed for serological analyses. The analysis of pooled samples instead of individual ones is a common procedure that permits to certify, with one single test, entire herds as "disease-free". However, diagnostic tests for pooled samples need to be particularly sensitive, especially when the levels of disease markers are low, as in the case of anti-BoHV1 antibodies in milk as markers of Infectious Bovine Rhinotracheitis (IBR) disease. The avidin-nucleic-acid-nanoassembly (ANANAS) is a novel kind of signal amplification platform for immunodiagnostics based on colloidal poly-avidin nanoparticles that, using model analytes, was shown to strongly increase ELISA test performance as compared to monomeric avidin. Here, for the first time, we applied the ANANAS reagent integration in a real diagnostic context. The monoclonal 1G10 anti-bovine IgG1 antibody was biotinylated and integrated with the ANANAS reagents for indirect IBR diagnosis from pooled milk mimicking tank samples from herds with IBR prevalence between 1 to 8%. The sensitivity and specificity of the ANANAS integrated method was compared to that of a classic test based on the same 1G10 antibody directly linked to horseradish peroxidase, and a commercial IDEXX kit recently introduced in the market. ANANAS integration increased by 5-fold the sensitivity of the 1G10 mAb-based conventional ELISA without loosing specificity. When compared to the commercial kit, the 1G10-ANANAS integrated method was capable to detect the presence of anti-BHV1 antibodies from bulk milk of gE antibody positive animals with 2-fold higher sensitivity and similar specificity. The results demonstrate the potentials of this new amplification technology, which permits improving current classic ELISA sensitivity limits

  12. A New ELISA Using the ANANAS Technology Showing High Sensitivity to diagnose the Bovine Rhinotracheitis from Individual Sera to Pooled Milk.

    Directory of Open Access Journals (Sweden)

    Elisabetta Casarin

    Full Text Available Diagnostic tests for veterinary surveillance programs should be efficient, easy to use and, possibly, economical. In this context, classic Enzyme linked ImmunoSorbent Assay (ELISA remains the most common analytical platform employed for serological analyses. The analysis of pooled samples instead of individual ones is a common procedure that permits to certify, with one single test, entire herds as "disease-free". However, diagnostic tests for pooled samples need to be particularly sensitive, especially when the levels of disease markers are low, as in the case of anti-BoHV1 antibodies in milk as markers of Infectious Bovine Rhinotracheitis (IBR disease. The avidin-nucleic-acid-nanoassembly (ANANAS is a novel kind of signal amplification platform for immunodiagnostics based on colloidal poly-avidin nanoparticles that, using model analytes, was shown to strongly increase ELISA test performance as compared to monomeric avidin. Here, for the first time, we applied the ANANAS reagent integration in a real diagnostic context. The monoclonal 1G10 anti-bovine IgG1 antibody was biotinylated and integrated with the ANANAS reagents for indirect IBR diagnosis from pooled milk mimicking tank samples from herds with IBR prevalence between 1 to 8%. The sensitivity and specificity of the ANANAS integrated method was compared to that of a classic test based on the same 1G10 antibody directly linked to horseradish peroxidase, and a commercial IDEXX kit recently introduced in the market. ANANAS integration increased by 5-fold the sensitivity of the 1G10 mAb-based conventional ELISA without loosing specificity. When compared to the commercial kit, the 1G10-ANANAS integrated method was capable to detect the presence of anti-BHV1 antibodies from bulk milk of gE antibody positive animals with 2-fold higher sensitivity and similar specificity. The results demonstrate the potentials of this new amplification technology, which permits improving current classic ELISA

  13. Quantification of bovine milk protein composition and coagulation properties using infrared spectroscopy and chemometrics: A result of collinearity among reference variables.

    Science.gov (United States)

    Eskildsen, C E; Skov, T; Hansen, M S; Larsen, L B; Poulsen, N A

    2016-10-01

    Predicting protein fractions and coagulation properties in bovine milk using Fourier transform infrared (FT-IR) measurements is desirable. However, such predictions may rely on correlations with total protein content. The aim of this study was to show how correlations between total protein content, protein fractions, and coagulation properties are responsible for the successful prediction of protein fractions and rennet-induced coagulation properties in milk samples. This study comprised 832 bovine milk samples from 2 breeds (426 Holstein and 406 Jersey). Holstein samples were collected from 20 Danish dairy herds from October to December 2009; Jersey samples were collected from 22 Danish dairy herds from February to April 2010. All samples were from conventional herds and taken while cows were housed. The results showed that κ-CN, αS1-CN, αS1-CN with 8 phosphorylated groups attached (αS1-CN 8P), and curd firming rate could be predicted from FT-IR measurements of the milk samples (with coefficients of determination between 0.66 and 0.71). However, the success of these FT-IR-based predictions was based on indirect relationships with total protein content. Hence, the FT-IR predictions relied on covariance structures with total protein content rather than absorption bands directly associated with the protein fractions and coagulation properties. If covariance structures between the protein fractions, coagulation properties, and total protein content used to calibrate partial least squares models were not conserved in future samples, these samples would show incorrect predictions of the protein fractions and coagulation properties. We demonstrated this using samples from 1 breed to calibrate and samples from the other breed to validate partial least squares models for β-CN. The 2 breeds had different covariance structures between β-CN and total protein content, and the validation samples yielded incorrect predictions. This finding may limit the usefulness of FT

  14. A triad of highly divergent polymeric immunoglobulin receptor (PIGR haplotypes with major effect on IgA concentration in bovine milk.

    Directory of Open Access Journals (Sweden)

    Sarah Berry

    Full Text Available The aim of this study was to determine a genetic basis for IgA concentration in milk of Bos taurus. We used a Holstein-Friesian x Jersey F2 crossbred pedigree to undertake a genome-wide search for QTL influencing IgA concentration and yield in colostrum and milk. We identified a single genome-wide significant QTL on chromosome 16, maximising at 4.8 Mbp. The polymeric immunoglobulin receptor gene (PIGR was within the confidence interval of the QTL. In addition, mRNA expression analysis revealed a liver PIGR expression QTL mapping to the same locus as the IgA quantitative trait locus. Sequencing and subsequent genotyping of the PIGR gene revealed three divergent haplotypes that explained the variance of both the IgA QTL and the PIGR expression QTL. Genetic selection based on these markers will facilitate the production of bovine herds producing milk with higher concentrations of IgA.

  15. Residues investigation of fluoroquinolones and sulphonamides and their metabolites in bovine milk by quantification and confirmation using ultra-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Meng, Zhe; Shi, Zhihong; Liang, Shuxuan; Dong, Xinfeng; Li, Hui; Sun, Hanwen

    2015-05-01

    A novel UPLC-MS/MS method for quantification and confirmation of eight fluoroquinolones, five sulphonamides (SAs) and four acetyled metabolites in milk is developed. Their main fragmentation pathways were presented. The limits of quantification were in the range of 0.01-0.29 μg kg(-1) for FQs and 0.13-1.68 μg kg(-1) for SAs. Mean recoveries ranging from 61% to 115% were achieved at spiked level of 0.5-100 μg kg(-1). The intra- and inter-day precisions were in the range of 6.3-10.7% and 9.0-13.4%, respectively. This novel approach has high speed and sensitivity, and can ensure the multiclass and multiresidue analysis of bovine milk at low μg kg(-1) level. The content of each analyte residue is much lower than the maximum residue limits. Some residues in milk were confirmed in according to the Commission Decision 657/2002/EC.

  16. Bovine milk intolerance in celiac disease is related to IgA reactivity to alpha- and beta-caseins.

    Science.gov (United States)

    Cabrera-Chávez, Francisco; de la Barca, Ana María Calderón

    2009-06-01

    Celiac disease is an autoimmune disease triggered mainly by ingestion of wheat gluten proteins. However, some other dietary proteins, such as those of cow's milk, induce celiac disease-like symptoms in some patients with celiac disease. Different approaches have been done to detect the component responsible for this problem, including the possibility of gluten peptides present in cow's milk.

  17. Computational analysis of bovine milk exosomal miRNAs profiles derived from uninfected and Streptococcus uberis infected mammary gland

    Science.gov (United States)

    The dairy cattle industry in the U.S. contributes an estimated 7 billion dollars to the agribusiness economy. Bacterial infections that cause disease like mastitis, affect health of the lactating mammary gland, and negatively impacts milk production and milk quality, costing producers an estimated 2...

  18. In vitro anti-inflammatory and xanthine oxidase inhibitory activity of Tephrosia purpurea shoot extract.

    Science.gov (United States)

    Nile, Shivraj H; Khobragade, Chandrahasy N

    2011-10-01

    The methanolic extract of Tephrosia purpurea (Leguminosae) shoots was evaluated in-vitro for its anti-inflammatory and xanthine oxidase inhibitory activity. Anti-inflammatory activity was measured by the Diene-conjugate, HET-CAM and beta-glucuronidase methods. The enzyme inhibitory activity was tested against isolated cow milk xanthine oxidase. The average anti-inflammatory activity of T. purpurea shoot extract in the concentration range of 1-2 microg/mL in the reacting system revealed significant anti-inflammatory activities, which, as recorded by the Diene-conjugate, HET-CAM and beta-glucuronidase assay methods, were 45.4, 10.5, and 70.5%, respectively. Screening of the xanthine oxidase inhibitory activity of the extract in terms of kinetic parameters revealed a mixed type of inhibition, wherein the Km and Vmax values in the presence of 25 to 100 microg/mL shoot extract was 0.20 mM/mL and 0.035, 0.026, 0.023 and 0.020 microg/min, while, for the positive control, the Km and Vmax values were 0.21 mM/mL and 0.043 microg/min, respectively. These findings suggest that T. purpurea shoot extract may possess constituents with good medicinal properties that could be exploited to treat the diseases associated with oxidative stress, xanthine oxidase enzyme activity and inflammation.

  19. Associations of a polymorphic AP-2 binding site in the 5'-flanking region of the bovine beta-lactoglobulin gene with milk proteins.

    Science.gov (United States)

    Kuss, A W; Gogol, J; Geidermann, H

    2003-06-01

    Studies on a polymorphic position (R10) in an Activator-Protein-2 (AP-2) binding site of the bovine beta-Lactoglobulin (beta-Lg) gene promoter region and quantitative traits of individual milk proteins were based on material from 79 German Holstein Friesian (HF) and 61 Simmental (Sm) cows. At least four milk samples per cow were analyzed with alkaline Urea-PAGE in combination with densitometry for quantification of individual milk proteins. The two alleles of the R10 single nucleotide polymorphism (SNP) carry either G or C in position -435 bp of the beta-Lg promoter region. G- and C-alleles were found in Sm with nearly equal frequencies, while in HF the C-allele frequency was higher (0.73) than that of the G-allele. In both breeds, the R10 G-homozygotes had higher (P beta-Lg secreted per day and proportion of beta-Lg in milk protein compared with the C-homozygotes. A similar association was found for alpha-lactalbumin, whereas the relative proportions and daily secreted amounts of caseins (alphaS1, beta, kappa) showed lower values in beta-Lg R10 G-homozygotes. A positive association (P beta-Lg locus to a candidate gene for this trait. The association between the SNP in the AP-2 binding site of the beta-Lg gene and its gene product can be explained as the result of differences in protein binding activity, and, therefore, allele specific differences in gene expression. Thus, our study clearly links a DNA polymorphism of molecular function very closely with in vivo expression parameters of the same locus.

  20. Enterotoxin Gene Profile and Molecular Characterization of Staphylococcus aureus Isolates from Bovine Bulk Milk and Milk Products of Tigray Region, Northern Ethiopia.

    Science.gov (United States)

    Tarekgne, Enquebaher K; Skjerdal, Taran; Skeie, Siv; Rudi, Knut; Porcellato, Davide; Félix, Benjamin; Narvhus, Judith A

    2016-08-01

    Staphylococcal food poisoning (SFP) is an important foodborne disease worldwide, and milk and milk products are commonly associated with SFP outbreaks. The objectives of this study were to investigate the distribution of staphylococcal enterotoxin (se) genes in Staphylococcus aureus from raw cow's milk and milk products and to assess their genetic background with the spa typing method. Of the 549 samples (297 bulk milk and 162 milk product samples) collected from Tigray region, Northern Ethiopia, 160 (29.1%) were positive for S. aureus, of which 82 (51%) were found to harbor se genes by a modified multiplex PCR. Nine se genes were identified: sea (n = 12), seb (n = 3), sec (n = 3), sed(n = 4), seg (n = 49), seh (n = 2), sei (n = 40), sej (n = 1), and tsst-1 (n = 24). The classical type of genes accounted for 27%. Of the 82 enterotoxigenic isolates, 41.5 and 12.4% harbored two or more se genes, respectively. The highest gene association was observed between sei and seg, whereas sea and seb were always found together with the new types of se genes. Altogether, 18 genotypes of toxin genes were identified, and 33% of the samples contained > 5 log CFU ml(-1) S. aureus. spa typing identified 22 spa types and three novel spa sequences, which showed the high genetic diversity of the isolates. No apparent relationship was observed between spa type and se genes. Of the 25 spa types, 13 (52%) were from raw milk, 3 (12%) from milk products, and 9 (36%) from both types of sample. Types t314 (20.7%,n = 17), t458 (18.3%, n = 15), and t6218 (9.8%, n= 8) were the most common spa types identified and were widely distributed in three of the eight studied localities. This is the first study from the Tigray region to report the high distribution of enterotoxigenic S. aureus with a diversified genetic background from dairy food. The study may provide valuable data for microbial food safety risk assessment, molecular epidemiology, and phylogenetic studies of S. aureus in Ethiopia.

  1. Genetic analysis of the Fourier-transform infrared spectra of bovine milk with emphasis on individual wavelengths related to specific chemical bonds.

    Science.gov (United States)

    Bittante, G; Cecchinato, A

    2013-09-01

    Fourier-transform infrared (FTIR) spectra are used to predict the fat, protein, casein, and lactose contents of milk. These estimates are currently used to predict the individual estimated breeding values of animals. The objective of the present study was to estimate the genetic variation and heritabilities of the milk transmittance spectrum at each individual FTIR wave. Milk was sampled once per cow from a total of 1,064 Italian Brown Swiss cows from 30 herds, sired by 50 artificial insemination sires. The FTIR spectra of all samples were collected within 3 h of sampling from 25 mL of milk. The obtained spectral range comprised wavenumbers 5,000 to 930×cm(-1), corresponding to wavelengths 2.00 to 10.76 μm and frequencies from 149.9 to 27.9 THz, for a total of 1,056 waves. These were acquired using a MilkoScan FT120 FTIR interferometer (Foss Electric A/S, Hillerød, Denmark). Each spectral data point was treated as a single trait and analyzed using an animal model REML method. The results indicated that the transmittance of the bovine milk FTIR spectrum was heritable for most individual waves in the wavenumber interval from 5,000 to 930×cm(-1). Moreover, the transmittance of contiguous FTIR waves was much more highly correlated in terms of the average value and phenotypic variation, compared with genetic variation. In the present study, we characterized 5 regions of the FTIR spectrum that were relevant to the analysis of milk; 2 regions, one in the transition area between the short-wavelength infrared (SWIR) and mid-wavelength infrared (MWIR) divisions of the electromagnetic spectrum (SWIR-MWIR region) and another very short region in the MWIR division (MWIR-2 region), were characterized by very high phenotypic variability in the transmittance of individual milk samples within each wave. This was caused by the absorption peaks of water, which can mask the effects of other important milk components. These regions also showed high genetic variability in

  2. Real-time polymerase chain reaction-based identification of bacteria in milk samples from bovine clinical mastitis with no growth in conventional culturing.

    Science.gov (United States)

    Taponen, S; Salmikivi, L; Simojoki, H; Koskinen, M T; Pyörälä, S

    2009-06-01

    In more than 30% of milk samples from clinical and subclinical bovine mastitis, bacteria fail to grow even after 48 h of conventional culture. The "no-growth" samples are problematic for mastitis laboratories, veterinarians, and dairy producers. This study provides the first investigation of the bacteriological etiology of such samples, using a real-time PCR-based commercial reagent kit. The assay targets the DNA of the 11 most common bacterial species or groups in mastitis and the staphylococcal blaZ gene (responsible for penicillin resistance) and can identify and quantify bacterial cells even if dead or growth-inhibited. A study was made of 79 mastitic milk samples with no-growth bacteria in conventional culture, originating from cows with clinical mastitis. Of the 79 samples, 34 (43%) were positive for 1 (32 samples) or 2 (2 samples) of the target bacteria. The positive findings included 11 Staphylococcus spp. (staphylococci other than Staphylococcus aureus), 10 Streptococcus uberis, 2 Streptococcus dysgalactiae, 6 Corynebacterium bovis, 3 Staph. aureus, 1 Escherichia coli, 1 Enterococcus, and 1 Arcanobacterium pyogenes. The positive samples contained as many as 10(3) to 10(7) bacterial genome copies per milliliter of milk. This study demonstrates that in nearly half of the clinical mastitis cases in which conventional culture failed to detect bacteria, mastitis pathogens were still present, often in substantial quantities. The clearly elevated N-acetyl-beta-d-glucosaminidase activity values of the milk samples, together with clinical signs of the infected cows and quarters, confirmed the diagnosis of clinical mastitis and indicated that real-time, PCR-based bacterial findings are able to reveal bacteriological etiology. We conclude that all common mastitis bacteria can occur in large quantities in clinical mastitis samples that exhibit no growth in conventional culture, and that the real-time PCR assay is a useful tool for bacteriological diagnosis of such

  3. The glycemic, insulinemic and plasma amino acid responses to equi-carbohydrate milk meals, a pilot- study of bovine and human milk

    DEFF Research Database (Denmark)

    Gunnerud, Ulrika; Holst, Jens Juul; Östman, Elin

    2012-01-01

    Dairy proteins, in particular the whey fraction, exert insulinogenic properties and facilitate glycemic regulation through a mechanism involving elevation of certain plasma amino acids, and stimulation of incretins. Human milk is rich in whey protein and has not been investigated in this respect....

  4. Bayesian estimation of sensitivity and specificity of Coxiella burnetii antibody ELISA tests in bovine blood and milk

    DEFF Research Database (Denmark)

    Paul, Suman; Toft, Nils; Agerholm, Jørgen S.;

    2013-01-01

    Serological tests for Coxiella burnetii (the causative agent of Q fever) antibodies are usually based on enzyme linked immunosorbent assay (ELISA) although this method is not thoroughly evaluated. The objective of this study was to determine the sensitivity and specificity of an ELISA for detection...... of the ELISA methods on milk and blood were equal at 0.99. No conditional dependence was observed between the specificity estimates of the two test methods. However, the sensitivity estimates of both tests were significantly reduced when conditional covariances ≥40 were used. Collection of milk samples from...... to positive (S/P) cut-off of 40 for both blood and milk ELISAs. At this cut-off, sensitivity of milk ELISA was 0.86 (95% posterior credibility interval [PCI] [0.76; 0.96]). This was slightly but insignificantly higher than sensitivity of blood ELISA (0.84; 95% PCI [0.75; 0.93]). The specificity estimates...

  5. 9 CFR 94.16 - Milk and milk products.

    Science.gov (United States)

    2010-01-01

    ... products, including dry whole milk, nonfat dry milk, dried whey, dried buttermilk, and formulations which... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Milk and milk products. 94.16 Section... VESICULAR DISEASE, AND BOVINE SPONGIFORM ENCEPHALOPATHY: PROHIBITED AND RESTRICTED IMPORTATIONS § 94.16...

  6. Determination of Quinolones and Nonsteroidal anti-Inflammatory Agents in Animal Tissues and Bovine Milk by Microwave-assisted Extraction High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    ZHOU Xi-Liu; DING Lan; JIN Hai-Yan; LIU Miao; CHENG Jian-Hua; WU Xiu-Feng; ZHAI Yu-Juan; SUN Yan-Tao; ZHANG Han-Qi; YU Yong; WANG Xiu-Pin

    2008-01-01

    A rapid,specific microwave-assisted extraction high performance liquid chromatography is described for assaying three quinolones(fleroxacin,lomefloxacin and sparfloxacin)and two nonsteroidal anti-inflammatory agents(ketoprofen and ibuprofen)in samples of sheep liver,bovine muscle and milk.The optimal microwave-assisted extraction conditions such as extraction temperature(40 ℃),extraction time(6 min),solvent volume(10 mL)and solvent(acetonitrile)were determined by an orthogonal experiment.Recoveries were 60.0%-107% in the concentration range 0.25-0.75 μg·g with good precision(< 11%)from three varieties of spiked animal samples.

  7. Complete characterization of posttranslational modification sites in the bovine milk protein PP3 by tandem mass spectrometry with electron capture dissociation as the last stage

    DEFF Research Database (Denmark)

    Kjeldsen, Frank; Haselmann, Kim F; Budnik, Bogdan A

    2003-01-01

    the PTM site. Chromatographic peak analysis continues until full sequence coverage is obtained, after which the molecular mass is reconstructed and compared with the measured value. An agreement indicates that the PTM characterization was complete. This procedure applied to the bovine milk PP3 protein......A comprehensive approach to protein identification and determination of sites of posttranslational modifications (PTMs) in heavily modified proteins was tested. In this approach, termed "reconstructed molecular mass analysis" (REMMA), the molecular mass distribution of the intact protein...... is measured first, which reveals the extent and heterogeneity of modifications. Then the protein is digested with one or several enzymes, with peptides separated by reversed-phase HPLC, and analyzed by Fourier transform mass spectrometry (FTMS). Vibrational excitation (collisional or infrared) or electron...

  8. In silico analysis of amino acid biosynthesis and proteolysis in Lactobacillus delbrueckii subsp. bulgaricus 2038 and the implications for bovine milk fermentation.

    Science.gov (United States)

    Zheng, Huajun; Liu, Enuo; Hao, Pei; Konno, Tomonobu; Oda, Munehiro; Ji, Zai-Si

    2012-08-01

    The amino acid biosynthesis pathway and proteolytic system of Lactobacillus delbrueckii subsp. bulgaricus 2038 (L. bulgaricus 2038), a mainstay of large-scale yogurt production, were modeled based on its genomic sequence. L. bulgaricus 2038 retains more potential for amino acid synthesis and a more powerful proteolytic system than other L. bulgaricus strains, but favors amino acid uptake over de novo synthesis. Free amino acids and peptides in bovine milk provide the main nitrogen sources; whey is more important than casein for L. bulgaricus during fermentation. Free amino acids are imported by amino acid permeases and by ABC-type transport systems whereas exogenous oligopeptides are imported by ABC-type proteins only. Histidine is neither synthesized nor imported singly, which might explain why L. bulgaricus cannot grow in synthetic media.

  9. 牛奶和奶粉中维吉尼霉素残留量的液相色谱——串联质谱测定%Determination of Virginiamycin Residue in Bovine Milk and Milk Powder by LC-MS-MS

    Institute of Scientific and Technical Information of China (English)

    赵静; 李广仁; 乔德福; 苏海滨; 陶迎春; 杨晓旎; 张晨; 宋文斌

    2013-01-01

    The virginiamycin M1 residues in bovine milk and milk powder were determined with LC-MS-MS.This method solved the determination of virginiamycin M1 residues in bovine milk and milk powder.The calibration curve of cirginiamycin M1 showed good linearity in the range of 0.125μg/kg-8.00μg/kg with r=0.9998.The detection limit was 0.25μg/kg for bovine milk and 2.00μg/kg for milk powder.Spiked with standard at four levels(n=10),the samples had the recoveries of 62.04%-111.65% for bovine milk and 85.49%-106.44% for milk powder,the relative standard deviation was 8.455%-20.03% for bovine milk and 2.467%-3.014% for milk powder.%建立了牛奶和奶粉中维吉尼霉素virginiamycin M1残留量的液相色谱——串联质谱测定方法.结果表明,维吉尼霉素M1在0.125μg/kg~8.00μg/kg范围内,具有良好的线性关系,相关系数r=0.9998.牛奶中维吉尼霉素M1方法检出限为0.25μg/kg,奶粉中维吉尼霉素M1方法检出限为2.00μg/kg.在4个添加水平上进行回收实验(n=10),牛奶平均回收率为62.04%~~ 111.65%,相对标准偏差在8.455%~20.03%之间;奶粉平均回收率为85.49%~106.44%,相对标准偏差在2.467%~3.014%之间.

  10. Molecularly imprinted polymer as in-line concentrator in capillary electrophoresis coupled with mass spectrometry for the determination of quinolones in bovine milk samples.

    Science.gov (United States)

    Moreno-González, David; Lara, Francisco J; Gámiz-Gracia, Laura; García-Campaña, Ana M

    2014-09-19

    In this work molecularly imprinted polymers have been evaluated as sorbent for the construction of an in-line solid phase extraction analyte concentrator in capillary electrophoresis coupled with mass spectrometry for the determination of the eight regulated veterinary quinolones in bovine milk samples. Different parameters affecting the analyte concentrator performance, such as sample pH, volume and composition of the elution plug and injection time, were studied. Sample volumes of 22μL (2bar for 15min) were loaded on the MISPE microcartridge and the retained analytes were eluted by injecting a plug of MeOH/H2O/NH3 (60/37/3 by volume) for 125s at 50mbar (60nL). The proposed method is simple for the monitoring of these antibiotic residues in milk samples, allowing the direct injection of the samples with minimum sample pretreatment, achieving limits of detection between 3.8 and 4.7μgkg(-1) and unequivocal identification of the compounds working in tandem mass spectrometry. Recoveries ranging from 70.0 to 102.3% were obtained and satisfactory intra-day and inter-day RSDs were achieved (≤12% and 15% respectively). Reproducibility among different constructed analyte concentrators showed RSD≤11%.

  11. Development and validation of a loop mediated isothermal amplification (LAMP) assay for the detection of Staphylococcus aureus in bovine mastitis milk samples.

    Science.gov (United States)

    Sheet, O H; Grabowski, N T; Klein, G; Abdulmawjood, A

    2016-10-01

    Staphylococcus (S.) aureus is one of the most important animal pathogens causing bovine mastitis. Also, it is a major human pathogen that may produce a variety of toxins which cause staphylococcal food poisoning. In the present study a LAMP assay based on gene nuc to identify S. aureus was developed and validated. The specificity of the LAMP assay was confirmed by using 70 S. aureus isolates and 21 non-S. aureus strains. The optimal temperature-time combination to amplify gene nuc successfully was 65 °C and 30 min. The analytical sensitivity of the developed LAMP assay was 0.26 pg of S. aureus DNA per reaction. The limit of detection evaluated with milk spiked with S. aureus was 9 × 10(2) CFU mL(-1). The final results of this assay were available within less than 2 h. The present study showed that the LAMP assay based on gene nuc appeared to be rapid and simple, and could also be used to identify S. aureus isolates from mastitis milk of dairy cows.

  12. High-throughput method for the determination of residues of β-lactam antibiotics in bovine milk by LC-MS/MS.

    Science.gov (United States)

    Jank, Louise; Martins, Magda Targa; Arsand, Juliana Bazzan; Hoff, Rodrigo Barcellos; Barreto, Fabiano; Pizzolato, Tânia Mara

    2015-01-01

    This study describes the development and validation procedures for scope extension of a method for the determination of β-lactam antibiotic residues (ampicillin, amoxicillin, penicillin G, penicillin V, oxacillin, cloxacillin, dicloxacillin, nafcillin, ceftiofur, cefquinome, cefoperazone, cephapirine, cefalexin and cephalonium) in bovine milk. Sample preparation was performed by liquid-liquid extraction (LLE) followed by two clean-up steps, including low temperature purification (LTP) and a solid phase dispersion clean-up. Extracts were analysed using a liquid chromatography-electrospray-tandem mass spectrometry system (LC-ESI-MS/MS). Chromatographic separation was performed in a C18 column, using methanol and water (both with 0.1% of formic acid) as mobile phase. Method validation was performed according to the criteria of Commission Decision 2002/657/EC. Main validation parameters such as linearity, limit of detection, decision limit (CCα), detection capability (CCβ), accuracy, and repeatability were determined and were shown to be adequate. The method was applied to real samples (more than 250) and two milk samples had levels above maximum residues limits (MRLs) for cloxacillin - CLX and cefapirin - CFAP.

  13. Milk supplemented with immune colostrum: protection against rotavirus diarrhea and modulatory effect on the systemic and mucosal antibody responses in calves experimentally challenged with bovine rotavirus.

    Science.gov (United States)

    Parreño, V; Marcoppido, G; Vega, C; Garaicoechea, L; Rodriguez, D; Saif, L; Fernández, F

    2010-07-01

    Group A bovine rotavirus (BRV) is the major cause of neonatal calf diarrhea worldwide. As a preventive strategy, we evaluated the protection and immunomodulation in two groups of BRV-inoculated calves. All calves received control colostrum (CC; VN=65,536; IgG(1)=16,384) prior to gut closure followed by the milk supplemented with immune colostrum (VN=1,048,576; IgG(1)=262,144), twice a day, for 14 days. Calves received milk supplemented with 0.8% immune colostrum [(Gp 1) VN=16,384; IgG(1)=4096] or milk supplemented with 0.4% immune colostrum [(Gp 2) VN=1024; IgG(1)=1024]. Calves receiving CC or colostrum deprived calves (CD) fed antibody (Ab) free milk served as controls (Gp 3 and 4). Calves were inoculated with virulent BRV IND at 2 days of age. Group 1 calves (milk IgG(1) 4096) showed 80% protection against BRV diarrhea and significantly reduced virus shedding. At 21 post-inoculation days (PID), the antibody secreting cell (ASC) responses of Gp 1 calves were limited mainly to duodenal and jejunal lamina propria (LP) with limited or no responses in systemic sites (spleen and PBL) and mesenteric lymph nodes. The profile of serum and fecal Ab responses as well as the ASC responses was also modulated by the presence of passive IgG(1) Abs and probably other colostrum components, toward higher titers of IgA Ab in serum and feces and a greater number of IgA ASC in the proximal intestine, reflecting positive modulation by colostrum toward this isotype associated with optimal protection of the intestinal mucosa. After challenge, at PID 21, all calves in Gp 1 and 2 were fully protected against diarrhea and only 1 of 5 calves in Gp 1 shed virus asymptomatically, indicating that the passive Ab treatment for 14 days was effective in protecting most of the animals after a first and a second virus exposure. The final outcome was a positive modulation of the mucosal immune responses and a high protection rate against diarrhea and virus shedding during the period of peak

  14. VASOACTIVE INTESTINAL POLYPEPTIDE PREVENTS INJURY OF PULMONARY VASCULAR PERMEABILITY DUE TO XANTHINE WITH XANTHINE OXIDASE

    Institute of Scientific and Technical Information of China (English)

    林耀广

    1995-01-01

    Hyperpermeability is a crux of pathogenesis of sudden lung edema in many pulmooary disorders,espe=cially in acute lung injury and adult respiratory distress syndrome(ARDS). Using our modified method for assessment of pulmonary vascular permeability,we observed the effects of xanthine with xanthine oxi-dase (X-XO) perfused in rat pulmonary artery and the protection of vasoactive intestinal polypeptide (VIP) against the injury of pulmonary vascular permesbility.After addition of xanthine oxidase in the perfusate reservoir containing xanthine,125I-albumin leak index (125IALI) was remarkably increased while peak airway pressure (Paw) was not significantly increased,and perfusion pressure of pulmonary artery (Ppa) and lung wet/dry weight ratio (W/D) were only slightly increased.Xanthine plus xanthine oxi-dase also increased thromboxane B2(TX B2) and 6-keto-prostaglandin F1α(6-keto-PGF1α) in the perfusat-e.Treatment with VIP obviously reduced or totally prevented all signs of injury.Simultaneously,VIP also diminished or abolished the associated generation of arachidonate products.The results indicated that VIP bas potent protective activity against injury of pulmonary vasculat permeability and may be a physiological modulator of inflammatory damage to vaseular eodothelium associated with toxic oxygen metabolites.

  15. Bovine Chymosin: A Computational Study of Recognition and Binding of Bovine κ-Casein

    DEFF Research Database (Denmark)

    Palmer, David S.; Christensen, Anders Uhrenholt; Sørensen, Jesper

    2010-01-01

    Bovine chymosin is an aspartic protease that selectively cleaves the milk protein κ-casein. The enzyme is widely used to promote milk clotting in cheese manufacturing. We have developed models of residues 97-112 of bovine κ-casein complexed with bovine chymosin, using ligand docking, conformational...

  16. Viral infections and bovine mastitis: a review

    NARCIS (Netherlands)

    Wellenberg, G.J.; Poel, van der W.H.M.; Oirschot, van J.T.

    2002-01-01

    This review deals with the role of viruses in the aetiology of bovine mastitis. Bovine herpesvirus 1, bovine herpesvirus 4, foot-and-mouth disease virus, and parainfluenza 3 virus have been isolated from milk from cows with clinical mastitis. Intramammary inoculations of bovine herpesvirus 1 or para

  17. Development of intramammary delivery systems containing lasalocid for the treatment of bovine mastitis: impact of solubility improvement on safety, efficacy, and milk distribution in dairy cattle

    Directory of Open Access Journals (Sweden)

    Wang W

    2015-01-01

    Full Text Available Wen Wang,1 Yunmei Song,1 Kiro Petrovski,2 Patricia Eats,2 Darren J Trott,2 Hui San Wong,2 Stephen W Page,3 Jeanette Perry,2 Sanjay Garg11School of Pharmacy and Medical Science, University of South Australia, Adelaide, SA, Australia; 2School of Animal and Veterinary Sciences, University of Adelaide, Adelaide, SA, Australia; 3Luoda Pharma Pty Ltd, Caringbah, NSW, AustraliaBackground: Mastitis is a major disease of dairy cattle. Given the recent emergence of methicillin-resistant Staphylococcus aureus as a cause of bovine mastitis, new intramammary (IMA treatments are urgently required. Lasalocid, a member of the polyether ionophore class of antimicrobial agents, has not been previously administered to cows by the IMA route and has favorable characteristics for development as a mastitis treatment. This study aimed to develop an IMA drug delivery system (IMDS of lasalocid for the treatment of bovine mastitis.Methods: Minimum inhibitory concentrations (MICs were determined applying the procedures recommended by the Clinical and Laboratory Standards Institute. Solid dispersions (SDs of lasalocid were prepared and characterized using differential scanning calorimetry and Fourier transform infrared spectroscopy. IMDSs containing lasalocid of micronized, nano-sized, or as SD form were tested for their IMA safety in cows. Therapeutic efficacy of lasalocid IMDSs was tested in a bovine model involving experimental IMA challenge with the mastitis pathogen Streptococcus uberis.Results: Lasalocid demonstrated antimicrobial activity against the major Gram-positive mastitis pathogens including S. aureus (MIC range 0.5–8 µg/mL. The solubility test confirmed limited, ion-strength-dependent water solubility of lasalocid. A kinetic solubility study showed that SDs effectively enhanced water solubility of lasalocid (21–35-fold. Polyvinylpyrrolidone (PVP-lasalocid SD caused minimum mammary irritation in treated cows and exhibited faster distribution in milk than

  18. Oral Microbiota in Infants Fed a Formula Supplemented with Bovine Milk Fat Globule Membranes - A Randomized Controlled Trial

    Science.gov (United States)

    Timby, Niklas; Domellöf, Magnus; Holgerson, Pernilla Lif; West, Christina E.; Lönnerdal, Bo; Hernell, Olle; Johansson, Ingegerd

    2017-01-01

    Background In a recent study, supplementation of infant formula with milk fat globule membranes (MFGM) decreased the incidence of otitis media in infants effects on the oral microbiome. Moraxella catarrhalis was less prevalent in infants fed EF than in those fed SF and may be associated with the decrease in otitis media seen in the same group. PMID:28099499

  19. Verification of fresh grass feeding, pasture grazing and organic farming by FTIR spectroscopy analysis of bovine milk

    NARCIS (Netherlands)

    Capuano, E.; Rademaker, J.; Bijgaart, van den H.; Ruth, van S.M.

    2014-01-01

    In the present study, a total of 116 tank milk samples were collected from 30 farms located in The Netherlands and analysed by Fourier-transform infrared (FTIR) spectroscopy. Samples were collected in April, May and June 2011 and in February 2012. The samples differed in the time spent by the cows o

  20. Detection and Enumeration of Streptococcus agalactiae from Bovine Milk Samples by Real-Time Polymerase Chain Reaction.

    Science.gov (United States)

    de Carvalho, Nara Ladeira; Gonçalves, Juliano Leonel; Botaro, Bruno Garcia; Silva, Luis Felipe de Prada E; dos Santos, Marcos Veiga

    2015-09-01

    The aim of this study was to evaluate the use of real-time polymerase chain reaction (qPCR) combined with DNA extraction directly from composite milk and bulk tank samples for detection and enumeration of Streptococcus agalactiae (SAG) causing subclinical mastitis. Dilutions of sterile reconstituted skim milk inoculated with SAG ATCC 13813 were used to establish a standard curve (cfu/mL) for the qPCR assay targeting SAG. The analytical sensitivity and repeatability of the qPCR assay were determined. Bulk tank (BTM; n = 38) and composite milk samples (CM; n = 26) collected from lactating cows with positive isolation of SAG were submitted to the qPCR protocol and SAG plate counting, with results from both methods compared. Amplification of DNA was not possible in two out of 64 samples, indicating that qPCR was able to detect SAG in 96 and 97% of BTM and CM samples, respectively. The inter-assay coefficient of variation was <5%, showing that the technique had adequate repeatability. The qPCR protocol can be a high-throughput and rapid diagnostic assay to accurately detect SAG from BTM and CM samples compared with conventional microbiological culture method. However, the evaluated qPCR protocol is not accurate for enumerating SAG in milk samples, probably due to quantification of DNA of non-viable cells.

  1. Analysis of relationship between bovine lymphocyte antigen DRB3.2 alleles, somatic cell count and milk traits in Iranian Holstein population.

    Science.gov (United States)

    Pashmi, M; Qanbari, S; Ghorashi, S A; Sharifi, A R; Simianer, H

    2009-08-01

    The major histocompatibility complex (MHC) is a gene complex closely linked to the vertebrate immune system due to its importance in antigen recognition and immune response to pathogens. To improve our understanding of the MHC and disease resistance in dairy cattle, we gathered 5119 test day records of somatic cell count (SCC) and performance traits of 262 Holstein dairy cows to determine whether the DRB region of the MHC contains alleles that are associated with elevated SCC, milk yield, protein and fat percent of milk. To this purpose, genotyping of animals for DRB3 gene was investigated by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) assay. A two-step PCR was carried out so as to amplify a 284 base-pair fragment of exon 2 of the target gene. Second PCR products were treated with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Twenty-eight BoLA-DRB3 alleles were identified including one novel allele (*40). The results in general are in good accordance with allele frequencies of Holstein cattle populations reported by previous studies. Analyses of associations were modeled based on repeated measurement anova and generalized logistic linear methods for production traits and SCC data, respectively. The results of this study showed a significant relationship between the elevated SCC reflecting an increased probability of occurrence to subclinical mastitis and DRB3.2 allele *8 (p < 0.03). The results also revealed significant positive relationships of alleles*22 (p < 0.01) and allele*11 (p < 0.05) with milk fat percent as well as of alleles*24 (p < 0.03) and *22 (p < 0.05) with protein percent. The present study failed to find any association between milk yield and tested alleles. Because of the lack of consistency among results of similar studies, we suggest further investigations to determine the precise nature of these associations with the high polymorphic bovine MHC region to be performed based on haplotypes.

  2. NMR-based milk metabolomics

    DEFF Research Database (Denmark)

    Sundekilde, Ulrik; Larsen, Lotte Bach; Bertram, Hanne Christine S.

    2013-01-01

    Milk is a key component in infant nutrition worldwide and, in the Western parts of the world, also in adult nutrition. Milk of bovine origin is both consumed fresh and processed into a variety of dairy products including cheese, fermented milk products, and infant formula. The nutritional quality...... and processing capabilities of bovine milk is closely associated to milk composition. Metabolomics is ideal in the study of the low-molecular-weight compounds in milk, and this review focuses on the recent nuclear magnetic resonance (NMR)-based metabolomics trends in milk research, including applications linking...... the milk metabolite profiling with nutritional aspects, and applications which aim to link the milk metabolite profile to various technological qualities of milk. The metabolite profiling studies encompass the identification of novel metabolites, which potentially can be used as biomarkers or as bioactive...

  3. A quantitative assessment of the risk of transmission of bovine spongiform encephalopathy by tallow-based calf milk-replacer.

    Science.gov (United States)

    Paisley, Larry G; Hostrup-Pedersen, Julie

    2004-04-30

    A Monte Carlo simulation model was constructed to assess the risk of BSE transmission to calves by calf milk-replacer (CMR). We assumed that any BSE infectivity in the CMR would be associated with the allowable levels of impurities in tallow used to manufacture the milk-replacer. Simulations used three different levels of impurities, six different distributions of the BSE infectivity titers of CNS tissues and with and without inclusion of specified risk material (SRM). Our results suggest that tallow-based CMR could have been responsible for some BSE infections in nearly all simulations. The reduction in the allowable impurities in tallow and the exclusion of SRM have greatly reduced--but have not eliminated--the risk of BSE transmission by CMR The results of the simulations are associated with much uncertainty.

  4. A quantitative assessment of the risk of transmission of bovine spongiform encephalopathy by tallow-based calf milk-replacer

    DEFF Research Database (Denmark)

    Paisley, Larry; Hostrup-Pedersen, J.

    2004-01-01

    A Monte Carlo simulation model was constructed to assess the risk of BSE transmission to calves by calf milk-replacer (CMR). We assumed that any BSE infectivity in the CMR would be associated with the allowable levels of impurities in tallow used to manufacture the milk-replacer. Simulations used...... three different levels of impurities, six different distributions of the BSE infectivity titers of CNS tissues and with and without inclusion of specified risk material (SRM). Our results suggest that tallow-based CMR could have been responsible for some BSE infections in nearly all simulations....... The reduction in the allowable impurities in tallow and the exclusion of SRM have greatly reduced-but have not eliminated-the risk of BSE transmission by CMR The results of the simulations are associated with much uncertainty....

  5. Fabrication of amperometric xanthine biosensors based on direct chemistry of xanthine oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Gao Yansheng; Shen Chunping [Department of Chemistry, Soochow University, Suzhou, Jiangsu 215123 (China); Di Junwei, E-mail: djw@suda.edu.cn [Department of Chemistry, Soochow University, Suzhou, Jiangsu 215123 (China); Tu Yifeng [Department of Chemistry, Soochow University, Suzhou, Jiangsu 215123 (China)

    2009-08-31

    The construction of amperometric xanthine biosensor by immobilization of xanthine oxidase (XOD) on the multi-wall carbon nanotubes (CNTs) modified glassy carbon (GC) electrode surface was investigated. The direct chemistry of XOD was accomplished and the formal potential was about - 0.465 V (vs SCE). The heterogeneous electron transfer rate constant was evaluated to be 2.0 {+-} 0.3 s{sup -1}. The xanthine biosensor based on XOD entrapped in silica sol-gel (SG) thin film on CNTs-modified GC electrode surface was also investigated. The XOD still maintains its activity to xanthine. The amperometric response to xanthine showed a linear relation in the range from 0.2 {mu}M to 10 {mu}M and a detection limit of 0.1 {mu}M (S/N = 3). The enzyme electrode retained 95% of its initial activity after 90 days of storage. The sensor exhibited high sensitivity, rapid response and good long-term stability.

  6. COMPARISON OF DIFFERENT SORTING OF FATTY ACID IN BOVINE MILK IN RELATION TO BODY CONDITION OF CZECH FLECKVIEH DAIRY COWS

    Directory of Open Access Journals (Sweden)

    Jaromir DUCHACEK

    2013-09-01

    Full Text Available Fatty acids in milk are one of the most important components of milk. The aim of this study was to determinate relationships between groups of fatty acids and body condition score change in Czech Fleckvieh cows. Fatty acids were classification along its length of chain and its source of creation. To analysis was included total of 50 Czech Fleckvieh cows with different order of lactation. During the first 4 weeks of lactation, milk samples were collected at a weekly interval and body condition score was assessed. Statistical analyses were performed using Microsoft Office Excel and the procedures MEANS and GLM of SAS 9.1. During the first four week of lactation, the proportions of short- and medium-chain fatty acids as well as de novo synthesised fatty acids increased. Moreover the cows with a greater body condition score change mobilized storage depot fat more intensively, which resulted in higher proportions of dietary and depot fatty acids since week 1 of lactation, and long-chain fatty acids since week 2 of lactation. On the contrary, the animals with only a small body condition score change exhibited high proportions of short- and medium-fatty acids as well as de novo synthesised fatty acids in the most part of the period analysed. This indicates abut negative energy balance in early part of lactation and its compensation. The results also confirm the relationships between different groups of fatty acids, body condition score and negative energy balance. As more accurate for evaluation of intensity of negative energy balance was confirmed rather classification along source of creation. These results further emphasize importance the monitoring of body condition, milk composition and good herd management in first part of lactation.

  7. Simultaneous quantification of carotenoids, retinol, and tocopherols in forage, bovine plasma, and milk: validation of a novel UPLC method

    Energy Technology Data Exchange (ETDEWEB)

    Chauveau-Duriot, B.; Doreau, M.; Noziere, P.; Graulet, B. [UR1213 Research Unit on Herbivores, INRA, Saint Genes Champanelle (France)

    2010-05-15

    Simultaneous quantification of various liposoluble micronutrients is not a new area of interest since these compounds participate in the nutritional quality of feeds that is largely explored in human, and also in animal diet. However, the development of related methods is still under concern, especially when the carotenoid composition is complex such as in forage given to ruminants or in lipid-rich matrices like milk. In this paper, an original method for simultaneous extraction and quantification of all carotenoids, vitamins E, and A in milk was proposed. Moreover, a new UPLC method allowing simultaneous determination of carotenoids and vitamins A and E in forage, plasma and milk, and separation of 23 peaks of carotenoids in forage was described. This UPLC method using a HSS T3 column and a gradient solvent system was compared to a previously published reverse-phase HPLC using two C18 columns in series and an isocratic solvent system. The UPLC method gave similar concentrations of carotenoids and vitamins A and E than the HPLC method. Moreover, UPLC allowed a better resolution for xanthophylls, especially lutein and zeaxanthin, for the three isomers of {beta}-carotene (all-E-, 9Z- and 13Z-) and for vitamins A, an equal or better sensitivity according to gradient, and a better reproducibility of peak areas and retention times, but did not reduce the time required for analysis. (orig.)

  8. Low-fat Gouda cheese made from bovine milk-olive oil emulsion: physicochemical and sensory attributes.

    Science.gov (United States)

    Felfoul, Imène; Bornaz, Salwa; Baccouche, Aroua; Sahli, Ali; Attia, Hamadi

    2015-10-01

    The objective of this paper is to study the effect of milk-fat replacement on Gouda cheeses composition, lipolysis and sensory characteristics. A Gouda cheese-like product was prepared from the substitution of milk fat with emulsified olive oil. For comparison, the low-fat variant without fat replacers and the full-fat cheese were also studied. Milk samples are initially pasteurized at 72 °C for 3 s, cooled to 35 °C, and added with 0.016 g L(-1) of lactic ferments and 0.30 mL L(-1) of microbial rennet. Total solids content was lower in cheeses containing fat replacers than in full and low-fat control cheeses due to the higher water-binding capacity of fat replacers. Free fatty acids rates were the highest in the case of reduced fat cheese-like product. The full-fat cheese showed a significantly higher overall impression score than all low-fat products.

  9. Electronic spectra and structures of some biologically important xanthines

    Science.gov (United States)

    Shukla, M. K.; Mishra, P. C.

    1994-08-01

    Electronic absorption and fluorescence spectra of aqueous solutions of xanthine, caffeine, theophylline and theobromine have been studied at different pH. The observed spectra have been interpreted in terms of neutral and ionic forms of the molecules with the help of molecular orbital calculations. At neutral and acidic pH, the spectra can be assigned to the corresponding most stable neutral forms, with the exception that the fluorescence of xanthine at acidic pH appears to originate from the lowest singlet excited state of a cation of the molecule. At alkaline pH, xanthine and theophylline exist mainly as their monoanions. In xanthine and theophylline at alkaline pH, fluorescence originates from the lowest singlet excited state of the corresponding anion. However, in caffeine and theobromine, even at alkaline pH, fluorescence belongs to the neutral species. On the whole, the properties of xanthine are quite different from those of the methyl xanthines.

  10. Transformation of serum-susceptible Escherichia coli O111 with p16Slux plasmid to allow for real-time monitoring of complement-based inactivation of bacterial growth in bovine milk.

    Science.gov (United States)

    Maye, S; Stanton, C; Fitzgerald, G F; Kelly, P M

    2016-01-01

    Complement activity has only recently been characterized in raw bovine milk. However, the activity of this component of the innate immune system was found to diminish as milk was subjected to heat or partitioning during cream separation. Detection of complement in milk relies on a bactericidal assay. This assay exploits the specific growth susceptibility of Escherichia coli O111 to the presence of complement. Practical application of the assay was demonstrated when a reduction in complement activity was recorded in the case of pasteurized and reduced-fat milks. This presented an opportunity to improve the functionality of the bactericidal assay by incorporating bioluminescence capability into the target organism. Following some adaptation, the strain was transformed by correctly integrating the p16Slux plasmid. Growth properties of the transformed strain of E. coli O111 were unaffected by the modification. The efficacy of the strain adaptation was correlated using the LINEST function analysis [r=0.966; standard error of prediction (SEy)=0.957] bioluminescence with that of bactericidal assay total plate counts within the range of 7.5 to 9.2 log cfu/mL using a combination of raw and processed milk samples. Importantly, the transformed E. coli O111 p16Slux strain could be identified in milk and broth samples using bioluminescence measurement, thus enabling the bactericidal assay-viability test to be monitored in real time throughout incubation.

  11. A retrospective evaluation of a Bovine Herpesvirus-1 (BHV-1) antibody ELISA on bulk-tank milk samples for classification of the BHV-1 status of Danish dairy herds

    DEFF Research Database (Denmark)

    Nylin, Britta; Strøger, Ulla; Rønsholt, Leif

    2000-01-01

    and relative specificity were 82 and 100%, respectively. The herd-level relative sensitivity depended on the within-herd prevalence of seropositive cows and the cut-off value in the assay, but not on the time interval (up to 90 days) between the collection of the bulk-tank milk sample and the individual serum......Bulk-tank milk samples analysed in a Bovine Herpesvirus-1 (BHV-1) blocking ELISA are still in use in the Danish BHV-1 programme as a tool to classify dairy herds as BHV-1 infected or BHV-1 free herds. in this retrospective study, we used data from the Danish BHV-1 eradication campaign to evaluate...... samples. The BHV-1 blocking ELISA on bulk-tank milk could detect seropositive herds (few), with prevalence proportions as low as one seropositive cow out of 70 cows....

  12. 用食品BDI-GS体系综合评价牛奶的营养健康效应%Comprehensive evaluation on nutrition and health of bovine milk using food BDI-GS system

    Institute of Scientific and Technical Information of China (English)

    龙伟; 沈秀; 王德芝; 王浩; 王晓光; 白佳利; 周则卫

    2013-01-01

    In this issue,study functions and safety of pure bovine milk through mice trial,establish a novel comprehensive system and method to evaluate nutrition,health-promotion and safety of food.The ICR male mice during growth were used in the trial,the experimental mice were feed with low nutrient maize and blending bovine milk feeds for 12 days,display and evaluate the effects of bovine milk with animal experiment data through observing and statistics the mice body weights,organ weights,organ indexes and its effect Benefit-damage Indexes (BDI).The study demonstrated that bovine milk possesses comprehensive and overall nutrition functions,certain health-promoting and foodtherapy functions for diabetes mellitus,and attenuating obesity effect,but some adverse response to bone mass.Bovine milk intake fits for nutrition of growing and nourishing for children and adolescents,nevertheless,drinking for a long period in large amount is not suitable for preventing adults and olds osteoporosis.The evaluation system has intuition,concrete,overall and comprehension features.This method can be used to evaluate nutrition,health-promotion and safety for various foods.%通过建立的食品功效安全BDI-GS评价体系和低营养小鼠模型方法综合评价纯牛奶的营养及健康效应.实验用生长期ICR雄性小鼠,通过12 d低营养玉米饲料和5%,10%及15%纯牛奶掺和饲料喂养,考察统计小鼠体质量、脏器质量、系数及其效应的损益指数(BDI)等指标,用动物实验数据评价牛奶功效.结果表明,牛奶的营养功效综合、全面;牛奶对II型糖尿病有一定的预防保健功效及控制体重(减肥)作用,但对骨质健康影响轻微.牛奶适合儿童、青少年生长发育饮用,适量饮用对中老年健康有益,而长期大量饮用对中老年防治骨质疏松不利.

  13. Preclinical investigations of a medium-chain triglyceride:fish oil emulsion. I. Effects of bovine milk lipoprotein lipase on lipid composition.

    Science.gov (United States)

    Carpentier, Yvon A; Dupont, Isabelle; Portois, Laurence; Malaisse, Willy J

    2006-10-01

    The bolus intravenous injection of a novel medium-chain triglyceride:fish oil emulsion (MCT:FO, 8:2, w:w) was recently found to increase within 60 min the leucocyte and platelet phospholipid content of long-chain polyunsaturated omega3 fatty acids. The present report deals with the effects of bovine milk lipoprotein lipase on the lipid composition of this emulsion. The results are compared to those obtained with either a pure fish oil emulsion or a medium-chain triglyceride: long-chain triglyceride:fish oil emulsion (MLF, 5:4:1). Emphasis is placed on i) differences in the fate of distinct fatty acids initially present in the triglycerides, di glycerides and phospholipids, ii) the generation of unesterified fatty acids relative to their initial content in each emulsion, and iii) the time course for these various events. The comparison between the three emulsions under consideration also provides information relevant to their respective sensitivity to lipoprotein lipase and suitability in terms of the generation of distinct unesterified fatty acids, including long-chain polyunsaturated omega3 fatty acids. Furthermore, attention is drawn to the greater efficiency for the hydrolysis of fatty acids from diglycerides as compared to triglycerides and a transient increase in the paired C8:0/C10:0 ratio in the diglycerides generated from the MCT:FO or MLF emulsion. The present study thus affords novel information relevant to the possible use of the MCT:FO emulsion in human subjects.

  14. Enterococcus faecalis strains from food, environmental, and clinical origin produce ACE-inhibitory peptides and other bioactive peptides during growth in bovine skim milk.

    Science.gov (United States)

    Gútiez, Loreto; Gómez-Sala, Beatriz; Recio, Isidra; del Campo, Rosa; Cintas, Luis M; Herranz, Carmen; Hernández, Pablo E

    2013-08-16

    Enterococcus faecalis isolates from food and environmental origin were evaluated for their angiotensin-converting enzyme (ACE)-inhibitory activity (ACE-IA) after growth in bovine skim milk (BSM). Most (90% active) but not all (10% inactive) E. faecalis strains produced BSM-derived hydrolysates with high ACE-IA. Known ACE-inhibitory peptides (ACE-IP) and an antioxidant peptide were identified in the E. faecalis hydrolysates by reversed-phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC-MS/MS). Antimicrobial activity against Pediococcus damnosus CECT4797 and Listeria ivanovii CECT913 was also observed in the E. faecalis hydrolysates. The incidence of virulence factors in the E. faecalis strains with ACE-IA and producers of ACE-IP was variable but less virulence factors were observed in the food and environmental strains than in the clinical reference strains. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) based analysis demonstrated that food and environmental E. faecalis strains were genetically different from those of clinical origin. When evaluated, most E. faecalis strains of clinical origin also originated BSM-derived hydrolysates with high ACE-IA due to the production of ACE-IP. Accordingly, the results of this work suggest that most E. faecalis strains of food, environmental and clinical origin produce BSM-derived bioactive peptides with human health connotations and potential biotechnological applications.

  15. β-lactam antibiotics residues analysis in bovine milk by LC-ESI-MS/MS: a simple and fast liquid-liquid extraction method.

    Science.gov (United States)

    Jank, L; Hoff, R B; Tarouco, P C; Barreto, F; Pizzolato, T M

    2012-01-01

    This study presents the development and validation of a simple method for the detection and quantification of six β-lactam antibiotics residues (ceftiofur, penicillin G, penicillin V, oxacillin, cloxacillin and dicloxacillin) in bovine milk using a fast liquid-liquid extraction (LLE) for sample preparation, followed by liquid chromatography-electrospray-tandem mass spectrometry (LC-MS/MS). LLE consisted of the addition of acetonitrile to the sample, followed by addition of sodium chloride, centrifugation and direct injection of an aliquot into the LC-MS/MS system. Separation was performed in a C(18) column, using acetonitrile and water, both with 0.1% of formic acid, as mobile phase. Method validation was performed according to the criteria of Commission Decision 2002/657/EC. Limits of detection ranged from 0.4 (penicillin G and penicillin V) to 10.0 ng ml(-1) (ceftiofur), and linearity was achieved. The decision limit (CCα), detection capability (CCβ), accuracy, inter- and intra-day repeatability of the method are reported.

  16. Calcium availability in bovine milk with treatment with gamma rays; Disponibilidade de calcio em leite bovino tratado por raios gama

    Energy Technology Data Exchange (ETDEWEB)

    Leonardi, Ana Lucia; Canniatti-Brazaca, Solange Guidolin, E-mail: leonardi@esalq.usp.b, E-mail: sgcbraza@esalq.usp.b [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Agroindustria, Alimentos e Nutricao; Arthur, Valter, E-mail: arthur@cena.usp.b [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil)

    2011-01-15

    Milk is an important food because it is rich in calcium and proteins. The objective of this research was to evaluate the effect of gamma radiation from {sup 60}Co on the calcium availability and sensorial aspects. The milk samples were packed in bags similar to those used for commercial products. The samples were submitted to gamma radiation at doses of 0.1, 0.5, 1.0, 2.0 and 3.0 kGy and stored at 4 deg C with evaluations on the 1st, 4th, 7th, and 10th days of storage. The calcium amount and centesimal composition were evaluated at the beginning of the study. Calcium dialysis in vitro was performed at different times of storage. Sensorial analysis was conducted on two doses with a more stable pH under the lowest radiation dose and the second lowest radiation dose, 0.5 and 1.0 kGy, respectively. Data analysis was conducted using the Tukey test (5%). Physicochemical analysis showed an increase in calcium availability, and the most adequate dose found was 1 kGy due to its highest pH stability in the period of 10 days, but the sensorial analysis showed that the samples were not well accepted by the testers. (author)

  17. Bovine Herpesvirus 4 infections and bovine mastitis

    NARCIS (Netherlands)

    Wellenberg, Gerardus Johannus

    2002-01-01

    Mastitis is an often occurring disease in dairy cattle with an enormous economic impact for milk producers worldwide. Despite intensive research, which is historically based on the detection of bacterial udder pathogens, still around 20-35% of clinical cases of bovine mastitis have an unknown aetiol

  18. A multidimensional high performance liquid chromatography method coupled with amperometric detection using a boron-doped diamond electrode for the simultaneous determination of sulfamethoxazole and trimethoprim in bovine milk

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, Leonardo S.; Moraes, Marcela C. de; Rocha-Filho, Romeu C.; Fatibello-Filho, Orlando [Departamento de Quimica, Universidade Federal de Sao Carlos, C.P. 676, 13560-970 Sao Carlos - SP (Brazil); Cass, Quezia B., E-mail: quezia@pesquisador.cnpq.br [Departamento de Quimica, Universidade Federal de Sao Carlos, C.P. 676, 13560-970 Sao Carlos - SP (Brazil)

    2009-11-10

    The development and validation of a multidimensional HPLC method using an on-line clean-up column coupled with amperometric detection employing a boron-doped diamond (BDD) electrode for the simultaneous determination of sulfamethoxazole (SMX) and trimethoprim (TMP) in bovine milk are presented. Aliquots of pre-prepared skim-milk samples were directly injected into a RAM octyl-BSA column in order to remove proteins that otherwise would interfere with milk analysis. After exclusion of the milk proteins, SMX and TMP were transferred to the analytical column (an octyl column) and the separation of the compounds from one another and from other endogenous milk components was achieved. SMX and TMP were detected amperometrically at 1.25 V vs. Ag/AgCl (3.0 mol L{sup -1} KCl). Results with good linearity in the concentration ranges 50-800 and 25-400 {mu}g L{sup -1} for SMX and TMP, respectively, were obtained and no fouling of the BDD electrode was observed within the experimental period of several hours. The intra- and inter-assay coefficients of variation were less than 10% for both drugs and the obtained LOD values for SMX and TMP were 25.0 and 15.0 {mu}g L{sup -1}, respectively.

  19. Development of a HS-SPME-GC-MS/MS Method for the Quantitation of Thymol and Carvacrol in Bovine Matrices and To Determine Residue Depletion in Milk and Tissues.

    Science.gov (United States)

    Armorini, Sara; Yeatts, James E; Mullen, Keena A E; Mason, Sharon E; Mehmeti, Elmira; Anderson, Kevin L; Washburn, Steve P; Baynes, Ronald E

    2016-10-11

    Thymol and carvacrol may be present in several phytoceutical products but there are no well-defined methods to measure these compounds in meat and milk from treated animals. U.S. regulatory authorities deem their presence as an adulteration of food. A rapid and sensitive HS-SPME-GC-MS/MS method was developed for the detection of thymol and carvacrol in bovine milk, plasma, liver, kidney, and fat. Inter- and intraday precision values were all less than 15.7 and 20.2% for thymol and carvacrol, respectively. The accuracy was in ranges of 69.9-111.8% for thymol and 74.0-119.2% for carvacrol. With the exception of fat tissue, stability studies showed that both compounds are stable over a 2 month period. A pilot pharmacokinetic study was conducted to evaluate the developed analytical method and to provide initial estimates of thymol and carvacrol depletion in plasma, milk, and several tissues. Treatment of lactating dairy cattle with phytoceutical products containing these substances resulted in low but measurable residue levels at 96 h for liver and 36 h for milk with very short apparent plasma and milk half-lives (<3.0 h).

  20. Detection of cow milk adulteration in yak milk by ELISA.

    Science.gov (United States)

    Ren, Q R; Zhang, H; Guo, H Y; Jiang, L; Tian, M; Ren, F Z

    2014-10-01

    In the current study, a simple, sensitive, and specific ELISA assay using a high-affinity anti-bovine β-casein monoclonal antibody was developed for the rapid detection of cow milk in adulterated yak milk. The developed ELISA was highly specific and could be applied to detect bovine β-casein (10-8,000 μg/mL) and cow milk (1:1,300 to 1:2 dilution) in yak milk. Cross-reactivity was detection limit was 1% (vol/vol) cow milk in yak milk. Different treatments, including heating, acidification, and rennet addition, did not interfere with the assay. Moreover, the results were highly reproducible (coefficient of variation detected no significant differences between known and estimated values. Therefore, this assay is appropriate for the routine analysis of yak milk adulterated with cow milk.

  1. The effect of the number of observations used for Fourier transform infrared model calibration for bovine milk fat composition on the estimated genetic parameters of the predicted data.

    Science.gov (United States)

    Rutten, M J M; Bovenhuis, H; van Arendonk, J A M

    2010-10-01

    Fourier transform infrared spectroscopy is a suitable method to determine bovine milk fat composition. However, the determination of fat composition by gas chromatography, required for calibration of the infrared prediction model, is expensive and labor intensive. It has recently been shown that the number of calibration samples is strongly related to the model's validation r(2) (i.e., accuracy of prediction). However, the effect of the number of calibration samples used, and therefore validation r(2), on the estimated genetic parameters of data predicted using the model needs to be established. To this end, 235 calibration data subsets of different sizes were sampled: n=100, n=250, n=500, and n=1,000 calibration samples. Subsequently, these data subsets were used to calibrate fat composition prediction models for 2 specific fatty acids: C16:0 and C18u (where u=unsaturated). Next, genetic parameters were estimated on predicted fat composition data for these fatty acids. Strong relationships between the number of calibration samples and validation r(2), as well as strong genetic correlations were found. However, the use of n=100 calibration samples resulted in a broad range of validation r(2) values and genetic correlations. Subsequent increases of the number of calibration samples resulted in narrowing patterns for validation r(2) as well as genetic correlations. The use of n=1,000 calibration samples resulted in estimated genetic correlations varying within a range of 0.10 around the average, which seems acceptable. Genetic analyses for the human health-related fatty acids C14:0, C16:0, and C18u, and the ratio of saturated fatty acids to unsaturated fatty acids showed that replacing observations on fat composition determined by gas chromatography by predictions based on infrared spectra reduced the potential genetic gain to 98, 86, 96, and 99% for the 4 fatty acid traits, respectively, in dairy breeding schemes where progeny testing is practiced. We conclude that

  2. Hydrolysis characteristics of bovine milk fat and monoacid triglycerides mediated by pregastric lipase from goats and kids.

    Science.gov (United States)

    Lai, D T; MacKenzie, A D; O'Connor, C J; Turner, K W

    1997-10-01

    Commercial extracts from oro-pharyngeal tissues of goats and kids have been used as the source of pregastric lipase and have been processed to yield partially purified samples of the primary pregastric lipase. The activity of these lipases against tributyrylglycerol has been determined over a range of pH and temperatures. Optimum pH conditions for pregastric lipase ranged from pH 5.6 to 6.5 for goats and from pH 5.5 to 6.2 for kids, respectively; the optimum temperature ranged from 43 to 60 degrees C. Optima for kid lipase extended slightly below pH 5.5 and higher than 60 degrees C; which were the limits of the test conditions. The enzymes were also used as catalysts for the hydrolysis of monoacid triglycerides (C4:0 to C12:0) at 40 degrees C and pH 6.5; activity was maximum against tributyrylglycerol (C4:0). Values for the Michaelis-Menten constant, increased as carbon chain length of the carboxylic moiety on the triglycerides increased, but values were identical for pregastric lipases of both goats and kids. Anhydrous milk fat was hydrolyzed by the commercial extracts of pregastric lipases of goats and kids, and the resulting profiles for free fatty acids were very similar to one another and to the corresponding profile for a commercial sample of Parmesan cheese. There appear to be no significant differences in activity between the enzyme preparations from goats and kids.

  3. Study of fodder, urine and bovine milk in marginal region of the Rio das Vellhas, Curvelo - Minas Gerais, Brazil; Estudo de forragem, urina e leite bovino na regiao ribeirinha do Rio da Velhas, Curvelo - Minas Gerais

    Energy Technology Data Exchange (ETDEWEB)

    Veado, Maria Adelaide R.V.; Oliveira, Arno H. de [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Dept. de Engenharia Nuclear]. E-mail: adelaide@nuclear.ufmg.br; heeren@nuclear.ufmg.br; fer@nuclear.ufmg.br; Veado, Julio Cesar C.; Melo, Marilia M. [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Escola de Veterinaria. Dept. de Clinica e Cirurgia Veterinarias]. E-mail: cambraia@vet.ufmg.br; Menezes, Maria Angela de B.C [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN), Belo Horizonte, MG (Brazil). Setor de Radioquimica. Supervisao de Reator e Radioanalise]. E-mail: menezes@urano.cdtn.br

    2000-07-01

    The parametric technique k{sub 0} of neutronic activation was used for determination of elements in samples of bovine urine and milk and fodder plant, of the region of farming production of Curvelo - Minas Gerais, Brazil. The samples had been collected in farms next to the margins to the Rio das Velhas that periodically suffer flooding in the rainy station. Clinical veterinarians studies disclose that many animals created in this region present illnesses with nervous symptomatology, suggestive of a picture of poisoning, however without definite diagnosis. These pathologies frequently are observed after the flooding. In this work had been determined the elements, Cr, Co, Zn among others. The results gotten in the different samples of urine and milk of the studied animals, as well as in the fodder plants, suggest that it has influence of loaded metals for waters of the high course of the basin river of Velhas.

  4. [Cow's milk protein allergy through human milk].

    Science.gov (United States)

    Denis, M; Loras-Duclaux, I; Lachaux, A

    2012-03-01

    Cow's milk protein allergy (CMPA) is the first allergy that affects infants. In this population, the incidence rate reaches 7.5%. The multiplicity and aspecificity of the symptoms makes its diagnosis sometimes complicated, especially in the delayed type (gastrointestinal, dermatological, and cutaneous). CMPA symptoms can develop in exclusively breastfed infants with an incidence rate of 0.5%. It, therefore, raises questions about sensitization to cow's milk proteins through breast milk. Transfer of native bovine proteins such as β-lactoglobulin into the breast milk is controversial: some authors have found bovine proteins in human milk but others point to cross-reactivity between human milk proteins and cow's milk proteins. However, it seems that a small percentage of dietary proteins can resist digestion and become potentially allergenic. Moreover, some authors suspect the transfer of some of these dietary proteins from the maternal bloodstream to breast milk, but the mechanisms governing sensitization are still being studied. Theoretically, CMPA diagnosis is based on clinical observations, prick-test or patch-test results, and cow's milk-specific IgE antibody concentration. A positive food challenge test usually confirms the diagnosis. No laboratory test is available to make a certain diagnosis, but the detection of eosinophil cationic protein (ECP) in the mother's milk, for example, seems to be advantageous since it is linked to CMA. Excluding cow's milk from the mother's diet is the only cure when she still wants to breastfeed. Usually, cow's milk proteins are reintroduced after 6 months of exclusion. Indeed, the prognosis for infants is very good: 80% acquire a tolerance before the age of 3 or 4 years. Mothers should not avoid dairy products during pregnancy and breastfeeding as preventive measures against allergy.

  5. Distinct composition of bovine milk from Jersey and Holstein-Friesian cows with good, poor or non-coagulation properties as reflected in protein genetic variants and isoforms

    DEFF Research Database (Denmark)

    Jensen, Hanne Bak; Poulsen, Nina Aagaard; Andersen, Kell Kleiner

    2012-01-01

    of minerals (Ca, P, Mg) were identified in poorly coagulating and noncoagulating milk in comparison with milk with good coagulation properties. Liquid chromatography/electrospray ionization-mass spectrometry revealed the presence of a great variety of genetic variants of the major milk proteins, namely, αS1...

  6. 营养素调控奶牛乳蛋白合成的研究进展%Recent Advance in Research on the Regulatory Effect of Nutrients on Bovine Milk Protein Synthesis

    Institute of Scientific and Technical Information of China (English)

    毕微微; 高学军; 林叶; 李庆章

    2012-01-01

    乳蛋白含有机体几乎所有的必需氨基酸,有较高的营养价值。随着蛋白质营养研究的深入,如何提高乳蛋白的产量,改善乳品质已成为当今研究的热点。本文从营养素角度,对碳水化合物、氨基酸及小肽三方面对奶牛乳蛋白合成调控的影响做综述。%Milk proteins contain nearly all the essential amino acids for the human body,which have great nutritional value.Today,how to increase milk protein production and improve milk quality has become the focus of increasingly intensive studies on protein nutrition.This paper provides a review of the regulatory effect of nutrients on bovine milk protein synthesis.

  7. 77 FR 15847 - Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products

    Science.gov (United States)

    2012-03-16

    ... been fed ruminant protein, other than milk protein, during their lifetime; The bovines from which the... from animals that are not known to have been fed ruminant protein, other than milk protein, during... community is that the agent is an abnormal form of a normal protein known as cellular prion protein. The...

  8. Cow's milk proteins in human milk.

    Science.gov (United States)

    Coscia, A; Orrù, S; Di Nicola, P; Giuliani, F; Rovelli, I; Peila, C; Martano, C; Chiale, F; Bertino, E

    2012-01-01

    Cow's milk proteins (CMPs) are among the best characterized food allergens. Cow's milk contains more than twenty five different proteins, but only whey proteins alpha-lactalbumin, beta-lactoglobulin, bovine serum albumin (BSA), and lactoferrin, as well as the four caseins, have been identified as allergens. Aim of this study was to investigate by proteomics techniques cow's milk allergens in human colostrum of term and preterm newborns' mothers, not previously detected, in order to understand if such allergens could be cause of sensitization during lactation. Term colostrum samples from 62 healthy mothers and preterm colostrum samples from 11 healthy mothers were collected for this purpose. The most relevant finding was the detection of the intact bovine alpha-S1-casein in both term and preterm colostrum. Using this method, which allows direct proteins identification, beta-lactoglobulin was not detected in any of colostrum samples. According to our results bovine alpha 1 casein that is considered a major cow's milk allergen is readily secreted in human milk: further investigations are needed in order to clarify if alpha-1-casein has a major role in sensitization or tolerance to cow's milk of exclusively breastfed predisposed infants.

  9. Evaluation of a commercial enzyme-linked immunosorbent assay for the diagnosis of bovine tuberculosis from milk samples from dairy cows.

    Science.gov (United States)

    Buddle, Bryce M; Wilson, Tania; Luo, Dongwen; Voges, Hinrich; Linscott, Richard; Martel, Edmond; Lawrence, John C; Neill, Mark A

    2013-12-01

    Milk samples from dairy cows provide a ready source of material for measuring antibody responses to Mycobacterium bovis antigens. In this study, we evaluated the IDEXX enzyme-linked immunosorbent assay (ELISA) for the measurement of antibody responses to M. bovis antigens MPB70 and MPB83 in milk samples from New Zealand cattle. Test sensitivities for individual milk and serum samples were assessed in samples collected from 44 M. bovis-infected cows, and test specificities were assessed in milk samples collected from 356 cows from tuberculosis (TB)-free herds. Milk vat samples were collected from 505 herds from regions with relatively high or low prevalences of infection. The ELISA had a sensitivity of 50% and a specificity of 97.5% for milk samples, and the test sensitivities for milk and serum samples were the same. Dilution of the positive test milk samples in milk from noninfected cows at 1/10, 1/20, and 1/50 dilutions reduced the proportions of positive responses to 13/21, 9/21, and 4/21, respectively. Small differences were observed in the ELISA responses of milk samples from individual TB-free cows collected at different times during lactation. No significant differences were detected in the ELISA responses of milk vat samples collected from infected and noninfected herds. This study shows that milk samples can be substituted for serum samples for screening individual cows for M. bovis infection, and pooling of milk samples from 10 to 20 animals can result in a reduction in the sensitivity by approximately 50%. However, screening of milk vat samples is unlikely to be useful in countries with low prevalences of M. bovis in cattle and large herd sizes.

  10. Método eletroforético rápido para detecção da adulteração do leite caprino com leite bovino Fast electrophoretic detection method of adulteration of caprine milk by bovine milk

    Directory of Open Access Journals (Sweden)

    A.S. Egito

    2006-10-01

    Full Text Available Avaliaram-se os métodos de eletroforese em gel de poliacrilamida (PAGE em presença de uréia (uréia-PAGE e dodecil sulfato de sódio (SDS-PAGE para identificar a adulteração do leite de cabra pela adição do leite de vaca. Um método foi otimizado para preparação do caseinato de sódio em poucos minutos para análise eletroforética. Uréia-PAGE foi o método mais apropriado para identificação desse tipo de fraude, em decorrência da presença da caseína alfas1 com migração mais rápida no leite bovino. A presença da alfas1-caseína bovina foi detectada a partir da adição de 2,5% de leite de vaca utilizando uréia-PAGE. O limite de detecção, a repetibilidade, o tempo para execução indicaram que esse método pode ser aplicado como rotina no controle de qualidade do leite de cabra recebido pelas indústrias de processamento.Polyacrylamide gel electrophoresis (PAGE in presence of urea (urea-PAGE or sodium dodecyl sulfate (SDS-PAGE was evaluated to detect the presence of cow milk added to goat milk. A method was optimized to prepare sodium caseinate from milk in few minutes. After that, the sodium caseinate was analyzed by PAGE. The urea-PAGE was the most appropriated method to identify adulteration as caprine and bovine alphas1-caseins displayed different migration rates. When cow milk was added to goat milk at different proportions, the presence of bovine alphas1-casein was detected in the mixture by urea-PAGE for a minimal proportion of 2.5% of cow milk added to goat milk. The good sensitivity, the repeatability and the short time for execution indicate that the described method will be able to be routinely applied for the quality control of goat milk in dairy industry.

  11. Kinetic study on the inhibition of xanthine oxidase by extracts from two selected Algerian plants traditionally used for the treatment of inflammatory diseases.

    Science.gov (United States)

    Berboucha, Meriem; Ayouni, Karima; Atmani, Dina; Atmani, Djebbar; Benboubetra, Mustapha

    2010-08-01

    In order to further understand and assess the validity of herbal medicine, we investigated the potential inhibitory effect of various extracts from Fraxinus angustifolia and Pistacia lentiscus, two plants used traditionally in Algeria against several inflammatory diseases such as rheumatism, arthritis, and gout, on purified bovine milk xanthine oxidase (XO) activity. The total phenolic contents of the leaves and bark of F. angustifolia and the leaves and seeds of P. lentiscus were estimated. P. lentiscus aqueous fractions from hexane and chloroform extractions and F. angustifolia aqueous fraction from ethyl acetate extraction inhibited XO activity by 72.74 +/- 2.63% (50% inhibitory concentration [IC(50)] = 27.52 microg/mL), 68.97 +/- 3.89% (IC(50) = 42.46 microg/mL) and 53.92 +/- 3.17% (IC(50) = 58.84 mmicroug/mL), respectively, at 100 microg/mL, compared to that of reference drug, allopurinol (98.18% [IC(50) = 6.34 microg/mL]). Moreover, at a concentration of 50 microg/mL, both P. lentiscus extracts showed inhibition rates higher than 50%. F. angustifolia leaf extracts showed only mild inhibition. Lineweaver-Burk analysis showed that the inhibitory activity exerted by F. angustifolia bark aqueous extract and P. lentiscus aqueous extracts is of mixed type, whereas the leaf extracts from F. angustifolia inhibited XO noncompetitively. Positive correlations were established between XO inhibition and total phenols (r = 0.89) and flavonoids (r = 0.93) for P. lentiscus and with total phenols (r = 0.72) and tannins (r = 0.54) for F. angustifolia. Our findings suggest that the therapeutic use of these plants may be due to the observed XO inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.

  12. Xanthine oxidase in human skeletal muscle following eccentric exercise

    DEFF Research Database (Denmark)

    Hellsten, Ylva; Frandsen, Ulrik; Orthenblad, N.

    1997-01-01

    1. The present study tested the hypothesis that the level of xanthine oxidase is elevated in injured human skeletal muscle in association with inflammatory events. Seven male subjects performed five bouts of strenuous one-legged eccentric exercise. Muscle biopsies from both the exercised and the ......1. The present study tested the hypothesis that the level of xanthine oxidase is elevated in injured human skeletal muscle in association with inflammatory events. Seven male subjects performed five bouts of strenuous one-legged eccentric exercise. Muscle biopsies from both the exercised...... the increase in xanthine oxidase in the muscle there were no detectable changes in the levels of muscle malondialdehyde or in plasma antioxidant capacity up to 4 days post-exercise. 5. It is concluded that eccentric exercise leads to an increased level of xanthine oxidase in human muscle and that the increase...

  13. Effect of Processing Temperature on Salts and Casein Colloid in Bovine Milk%热处理对牛乳中盐类组分及酪蛋白胶体特性的影响

    Institute of Scientific and Technical Information of China (English)

    王辉; 孙琦; 刘鹭; 张书文; 梁霄; 吕加平

    2012-01-01

    As important components of milk,salts and casein colloid play a key role in maintaining the stability of milk system.In order to explore the effect of processing temperature on salts and casein colloid in bovine milk,we measured indicators of milk quality at different temperatures.The results showed that processing temperature had a significant impact on the form and content of salts in milk(P0.05).As the temperature increased,the soluble calcium and soluble phosphorus were transferred to the colloidal phase;milk casein colloidal particle size,surface potential,hydration and other properties also revealed an obvious change due to the denaturation of whey protein.Therefore,there is a close relationship of processing temperature with salts and casein colloid in milk,which is important for the quality of milk.%为研究热处理对牛乳中盐类组分和酪蛋白胶体特性的影响,对不同温度处理后的牛乳进行相关指标的测定。结果表明:不同温度处理对牛乳中主要盐类的存在形态及含量有着显著影响(P〈0.05),随着温度的升高,溶解相的钙和磷逐渐向胶体相转变;同时,受乳清蛋白变性的影响,牛乳中酪蛋白胶体粒径、表面电势、水合作用等特性也发生了显著变化。因此,加工温度与牛乳中的盐类组分及酪蛋白胶体特性有着密切关联,是造成牛乳品质变化的一个重要因素。

  14. NMR-Based Milk Metabolomics

    Directory of Open Access Journals (Sweden)

    Hanne C. Bertram

    2013-04-01

    Full Text Available Milk is a key component in infant nutrition worldwide and, in the Western parts of the world, also in adult nutrition. Milk of bovine origin is both consumed fresh and processed into a variety of dairy products including cheese, fermented milk products, and infant formula. The nutritional quality and processing capabilities of bovine milk is closely associated to milk composition. Metabolomics is ideal in the study of the low-molecular-weight compounds in milk, and this review focuses on the recent nuclear magnetic resonance (NMR-based metabolomics trends in milk research, including applications linking the milk metabolite profiling with nutritional aspects, and applications which aim to link the milk metabolite profile to various technological qualities of milk. The metabolite profiling studies encompass the identification of novel metabolites, which potentially can be used as biomarkers or as bioactive compounds. Furthermore, metabolomics applications elucidating how the differential regulated genes affects milk composition are also reported. This review will highlight the recent advances in NMR-based metabolomics on milk, as well as give a brief summary of when NMR spectroscopy can be useful for gaining a better understanding of how milk composition is linked to nutritional or quality traits.

  15. A gene-based high-resolution comparative radiation hybrid map as a framework for genome sequence assembly of a bovine chromosome 6 region associated with QTL for growth, body composition, and milk performance traits

    Directory of Open Access Journals (Sweden)

    Laurent Pascal

    2006-03-01

    Full Text Available Abstract Background A number of different quantitative trait loci (QTL for various phenotypic traits, including milk production, functional, and conformation traits in dairy cattle as well as growth and body composition traits in meat cattle, have been mapped consistently in the middle region of bovine chromosome 6 (BTA6. Dense genetic and physical maps and, ultimately, a fully annotated genome sequence as well as their mutual connections are required to efficiently identify genes and gene variants responsible for genetic variation of phenotypic traits. A comprehensive high-resolution gene-rich map linking densely spaced bovine markers and genes to the annotated human genome sequence is required as a framework to facilitate this approach for the region on BTA6 carrying the QTL. Results Therefore, we constructed a high-resolution radiation hybrid (RH map for the QTL containing chromosomal region of BTA6. This new RH map with a total of 234 loci including 115 genes and ESTs displays a substantial increase in loci density compared to existing physical BTA6 maps. Screening the available bovine genome sequence resources, a total of 73 loci could be assigned to sequence contigs, which were already identified as specific for BTA6. For 43 loci, corresponding sequence contigs, which were not yet placed on the bovine genome assembly, were identified. In addition, the improved potential of this high-resolution RH map for BTA6 with respect to comparative mapping was demonstrated. Mapping a large number of genes on BTA6 and cross-referencing them with map locations in corresponding syntenic multi-species chromosome segments (human, mouse, rat, dog, chicken achieved a refined accurate alignment of conserved segments and evolutionary breakpoints across the species included. Conclusion The gene-anchored high-resolution RH map (1 locus/300 kb for the targeted region of BTA6 presented here will provide a valuable platform to guide high-quality assembling and

  16. Fatty acid composition, fat soluble vitamin concentrations and oxidative stability in bovine milk produced on two pastures with different botanical composition

    DEFF Research Database (Denmark)

    Adler, S A; Dahl, A V; Jensen, Søren Krogh

    2013-01-01

    that no artificial fertilizers or herbicides were applied. The SP was representative for pastures, which are frequently, i.e. at least every third year, renewed by soil tillage and seeding, whereas LP was representative for pastures, which are less frequently renewed. The SP contained mainly meadow fescue (Festuca...... liquid chromatography, and oxidative stability in a light-exposure experiment by measuring the formation of hydroperoxides and by front-face fluorescence spectroscopy. Pasture type had no effect on milk yield, milk gross composition, and only minor effects on milk FA composition. Milk from SP had higher...

  17. Problemática del arsénico en la llanura sudeste de la provincia de Córdoba: Biotransferencia a leche bovina Arsenic in the southeast plains of Cordoba province, Argentine: Biotransference to bovine milk

    Directory of Open Access Journals (Sweden)

    A Pérez Carrera

    2007-12-01

    Full Text Available La transferencia de elementos traza desde matrices ambientales a la cadena agroalimentaria es un fenómeno que está siendo estudiado en distintas regiones del mundo. Sin embargo, la información acerca de los niveles en alimentos, particularmente la leche, es escasa. En nuestro país, un elemento de particular relevancia es el arsénico (As, por su ocurrencia natural en aguas subterráneas en una amplia zona de nuestro territorio. En este trabajo se determinaron las concentraciones de As en agua de bebida animal, suelo, forraje y leche en establecimientos lecheros del sudeste de la provincia de Córdoba. Los niveles de As determinados en agua de bebida, en particular en aquellos establecimientos que extraían el agua de la capa freática, sugerían la posibilidad de transferencia de As a la leche bovina. Los niveles en suelo y forraje estuvieron dentro de los rangos de valores normales para regiones de abundancia natural de As. Las concentraciones de As determinadas en muestras de leche no revestían toxicidad para consumo humano. Los resultados obtenidos permitieron estimar un factor de biotransferencia (FBT de As a leche bovina a partir de la ingesta de este elemento por parte de los animales, en las condiciones productivas y edafoclimáticas de nuestro país.Trace element transference from environmental matrices to the agricultural food chain is an actual phenomenon that is being analyzed in different regions of the world. Nevertheless, knowledge about trace element content in food, particularly in milk, is scarce. In Argentine, an element of great concern is arsenic due to its natural occurrence in groundwater in large areas of the country. The aim of the present work was determine arsenic content in livestock drinking water and milk from dairy farms located in the southeast of the Cordoba province. Arsenic concentrations in all collected phreatic samples suggested the possibility of transference to bovine milk. Arsenic levels in soils and

  18. Effect of bovine colostrum feeding in comparison with milk replacer and natural feeding on the immune responses and colonisation of enterotoxigenic Escherichia coli in the intestinal tissue of piglets

    DEFF Research Database (Denmark)

    Sugiharto, Sugiharto; Poulsen, Ann-Sofie Riis; Canibe, Nuria;

    2015-01-01

    (23-d-old) were allocated to one of the following four groups: (1) killed at the beginning of the experiment (Base); (2) separated from the sow and fed BC (BC-fed); (3) separated from the sow and fed a MR (MR-fed); (4) kept with the sow (Sow-Milk). Blood was sampled on days 1 and 8, and faecal samples......The present study investigated the effect of feeding bovine colostrum (BC) to piglets in comparison with feeding a milk replacer (MR) and conventional rearing by the sow on the intestinal immune system and number of enterotoxigenic Escherichia coli (ETEC) colonising the intestinal tissue. Piglets...... were collected on days 1, 3, 5 and 8. On day 8, piglets were killed and gastrointestinal digesta and intestinal segments were collected. The frequency of diarrhoea was found to be higher (P≤ 0·019) in MR-fed piglets than in BC-fed and Sow-Milk piglets. Piglets from the MR-fed group had the lowest...

  19. Effect of fresh grass feeding, pasture grazing and organic/biodynamic farming on bovine milk triglyceride profile and implications for authentication

    NARCIS (Netherlands)

    Capuano, E.; Boerrigter-Eenling, G.R.; Eigersma, A.; Ruth, van S.M.

    2014-01-01

    In the present study, 113 tank milk samples were collected from 30 farms located in the Netherlands and analyzed for their triacylglycerol (TAG) and cholesterol profiles. Significant differences in the TAG profile between winter and spring–early summer milk were observed. The differences between mil

  20. Genome-wide scan for bovine milk-fat composition. I. Quantitative trait loci for short- and medium-chain fatty acids

    NARCIS (Netherlands)

    Stoop, W.M.; Schennink, A.; Visker, M.H.P.W.; Mullaart, E.; Arendonk, van J.A.M.; Bovenhuis, H.

    2009-01-01

    A genome-wide scan was performed to identify quantitative trait loci (QTL) for short- and medium-chain fatty acids (expressed in wt/wt %). Milk samples were available from 1,905 cows from 398 commercial herds in the Netherlands, and milk-fat composition was measured by gas chromatography. DNA was av

  1. Short communication: Genome-wide scan for bovine milk-fat composition. II. Quantitative trait loci for long-chain fatty acids

    NARCIS (Netherlands)

    Schennink, A.; Stoop, W.M.; Visker, M.H.P.W.; Poel, van der J.J.; Bovenhuis, H.; Arendonk, van J.A.M.

    2009-01-01

    We present the results of a genome-wide scan to identify quantitative trait loci (QTL) that contribute to genetic variation in long-chain milk fatty acids. Milk-fat composition phenotypes were available on 1,905 Dutch Holstein-Friesian cows. A total of 849 cows and their 7 sires were genotyped for 1

  2. Effect of carryover and presampling procedures on the results of real-time PCR used for diagnosis of bovine intramammary infections with Streptococcus agalactiae at routine milk recordings.

    Science.gov (United States)

    Mahmmod, Yasser S; Mweu, Marshal M; Nielsen, Søren S; Katholm, Jørgen; Klaas, Ilka C

    2014-03-01

    The use of PCR tests as diagnostics for intramammary infections (IMI) based on composite milk samples collected in a non-sterile manner at milk recordings is increasing. Carryover of sample material between cows and non-aseptic PCR sampling may be incriminated for misclassification of IMI with Streptococcus agalactiae (S. agalactiae) in dairy herds with conventional milking parlours. Misclassification may result in unnecessary costs for treatment and culling. The objectives of this study were to (1) determine the effect of carryover on PCR-positivity for S. agalactiae at different PCR cycle threshold (Ct) cut-offs by estimating the between-cow correlation while accounting for the milking order, and (2) evaluate the effect of aseptic presampling procedures (PSP) on PCR-positivity at the different Ct-value cut-offs. The study was conducted in four herds with conventional milking parlours at routine milk recordings. Following the farmers' routine pre-milking preparation, 411 of 794 cows were randomly selected for the PSP treatment. These procedures included removing the first streams of milk and 70% alcohol teat disinfection. Composite milk samples were then collected from all cows and tested using PCR. Data on milking order were used to estimate the correlation between consecutively milked cows in each milking unit. Factors associated with the PCR-positivity for S. agalactiae were analyzed using generalized estimating equations assuming a binomially-distributed outcome with a logit link function. Presampling procedures were only significant using cut-off 37. A first-order autoregressive correlation structure provided the best correlation between consecutively milked cows. The correlation was 13%, 11%, 9% at cut-offs <40, 37, and 34, respectively. PSP did not reduce the odds of cows being PCR-positive for S. agalactiae. In conclusion, carryover and non-aseptic sampling affected the PCR results and should therefore be considered when samples from routine milk

  3. Natural variation in casein composition of milk

    NARCIS (Netherlands)

    Bijl, E.

    2014-01-01

    Bovine milk contains 3-4 % protein and almost 80% of the milk protein fraction consist of four caseins; αs1-casein, β-casein, αs2-casein and κ-casein. Most of the caseins in milk are assembled in casein micelles, which consist of several thousands of indi

  4. 奶牛乳腺上皮细胞不同代数对乳蛋白基因表达的影响%Different Generations of Bovine Mammary Epithelial Cells Impacting on Gene Expression of Milk Protein

    Institute of Scientific and Technical Information of China (English)

    韩亚南; 侯先志; 考桂兰; 王秀美; 杨金丽; 高爱武; 杨银芬; 赵青; 高民

    2012-01-01

    The aim of this study was to research milk protein gene expression—αsl-casein, β-casein, κ-casein of lactating bovine mammary epithelial cells between different generations; Using the real—time PCR method to detect milk protein gene expression of lactating bovine mammary tissue, the cells of primary generation (P0) derived from the bovine mammary tissue, the first generation (Pi), the second generation (P2), the third generation (P3), the fourth generation (P4) and the fifth generation (P3); The results showed that expression of three target-genes was at different levels between the cells of primary generation and passages. The expression in the primary cells was significantly higher than P1-P5 cells (P<0.05). Whereas P1-P5 cells had no significant difference (P>0.05). Moreover, from Po to P5, the expression of αsl-casein, β-casein, K-casein was gradually decreasing. So, P2-P5 BMECs could be used as a trial material.%为了研究泌乳期奶牛乳腺上皮细胞(bovine mammary epithelial cells,BMECs)不同代数乳蛋白基因αs1-casein,β-casein,κ-casein的表达.利用real-time PCR方法分别对泌乳期奶牛乳腺组织,以及由乳腺组织分离得到的细胞原代(P0),1代(P1),2代(P2),3代(P3),4代(P4),5代(P5)中的乳蛋白基因表达状况进行检测.结果表明:αs1-casein,β-casein,κ-casein在泌乳期奶牛乳腺上皮细胞原代及传代细胞中有不同程度的表达.其中,原代中的表达量显著高于P1~P5(P<0.05),而P1~P5之间的表达量无显著差异(P>0.05).并且随着传代次数的增加,基因表达量呈现逐渐下降的趋势.说明P2~P5 BMECs可以作为基础理论问题研究的试验材料而被应用.

  5. Filter-aided sample preparation with dimethyl labeling to identify and quantify milk fat globule membrane proteins.

    NARCIS (Netherlands)

    Lu, J.; Boeren, J.A.; Vries, de S.C.; Valenberg, van H.J.F.; Vervoort, J.J.M.; Hettinga, K.A.

    2011-01-01

    Bovine milk is a major nutrient source in many countries and it is produced at an industrial scale. Milk is a complex mixture of proteins, fats, carbohydrates, vitamins and minerals. The composition of the bovine milk samples can vary depending on the genetic makeup of the bovine species as well as

  6. Investigation of Brucella Infection in Milk Collected from Burdur Province

    OpenAIRE

    TÜRÜTOĞLU, Hülya

    2014-01-01

    In this study, we aimed to investigate the regional profile of bovine and ovine Brucellosis in Burdur by isolating Brucella spp. in milk and by detecting antibodies to Brucellae in milk samples. For this purpose, 404 milk samples from 101 cows and 226 milk samples from 113 ewes located in 14 different areas of Burdur were collected. The samples were examined by bacteriological and serological methods. Brucella spp. were not isolated from the milk samples. Positive results by the milk ring te...

  7. Relationship between the metabolite profile and technological properties of bovine milk from two dairy breeds elucidated by NMR-based metabolomics.

    Science.gov (United States)

    Sundekilde, Ulrik Kræmer; Frederiksen, Pernille Dorthea; Clausen, Morten Rahr; Larsen, Lotte Bach; Bertram, Hanne Christine

    2011-07-13

    The aim of the present study was to investigate the relationship between the metabolite profile of milk and important technological properties by using nuclear magnetic resonance (NMR)-based metabolomics. The metabolomics approach was introduced for the metabolic profiling of a set of milk samples from two dairy breeds representing a wide span in coagulation properties. The milk metabolite profiles obtained by proton and carbon NMR spectroscopy could be correlated to breed and, more interestingly, also with the coagulation profile, as established by traditional methods by using principal component analysis (PCA). The metabolites responsible for the separation into breed could mainly be ascribed to carnitine and lactose, whereas the metabolites varying in the samples with respect to coagulation properties included citrate, choline, carnitine, and lactose. The results found in the present study demonstrated a promising potential of NMR-based metabolomics for a rapid analysis and classification of milk samples, both of which are useful for the dairy industry.

  8. The biology of milk synthesis from a proteomics perspective

    NARCIS (Netherlands)

    Lu, J.

    2013-01-01

    Large variation in bovine milk composition of Dutch Holstein cows has been observed. The factors influencing the milk synthesis and secretion process in the mammary gland and the variations in this process lead to variation in milk composition. The understanding of milk synthesis was improved during

  9. Xanthine oxidoreductase and its inhibitors: relevance for gout.

    Science.gov (United States)

    Day, Richard O; Kamel, Bishoy; Kannangara, Diluk R W; Williams, Kenneth M; Graham, Garry G

    2016-12-01

    Xanthine oxidoreductase (XOR) is the rate-limiting enzyme in purine catabolism and converts hypoxanthine to xanthine, and xanthine into uric acid. When concentrations of uric acid exceed its biochemical saturation point, crystals of uric acid, in the form of monosodium urate, emerge and can predispose an individual to gout, the commonest form of inflammatory arthritis in men aged over 40 years. XOR inhibitors are primarily used in the treatment of gout, reducing the formation of uric acid and thereby, preventing the formation of monosodium urate crystals. Allopurinol is established as first-line therapy for gout; a newer alternative, febuxostat, is used in patients unable to tolerate allopurinol. This review provides an overview of gout, a detailed analysis of the structure and function of XOR, discussion on the pharmacokinetics and pharmacodynamics of XOR inhibitors-allopurinol and febuxostat, and the relevance of XOR in common comorbidities of gout.

  10. Monitoring the freezing point of buffalo milk

    Directory of Open Access Journals (Sweden)

    Antonella Pesce

    2016-05-01

    Full Text Available The aim of this study was to evaluate the basic freezing point of buffalo milk. Bulk milk samples were collected from buffalo and cattle farms in Caserta area from 2008 to 2014. The analysis involved a total of 1886 buffalo milk samples and 1711 bovine milk samples. These were also tested for fat, protein and lactose contents by means of infrared spectrometry. The freezing point was determined by means of a thermistor cryoscope. Data underwent statistical analysis. Our research showed an average freezing point of -0.528°C for buffalo milk and - 0.522°C for bovine milk. Given the lack of data on the freezing point of buffalo milk, our study provides the first indication of a basic freezing point of the milk of this species in Italy.

  11. Milk: the new sports drink? A Review

    OpenAIRE

    Roy Brian D

    2008-01-01

    Abstract There has been growing interest in the potential use of bovine milk as an exercise beverage, especially during recovery from resistance training and endurance sports. Based on the limited research, milk appears to be an effective post-resistance exercise beverage that results in favourable acute alterations in protein metabolism. Milk consumption acutely increases muscle protein synthesis, leading to an improved net muscle protein balance. Furthermore, when post-exercise milk consump...

  12. Entrainment in solution of an oscillating NADH oxidase activity from the bovine milk fat globule membrane with a temperature-compensated period length suggestive of an ultradian time-keeping (clock) function

    Science.gov (United States)

    Morre, D. James; Lawler, Juliana; Wang, Sui; Keenan, Thomas W.; Morre, Dorothy M.

    2002-01-01

    Entrainment in solution of an oscillating activity with a temperature compensated period of 24 min is described for a NADH oxidase (NOX) activity of the bovine milk fat globule membrane, a derivative of the mammary epithelial cell plasma membrane. The period of 24 min remained unchanged at 17 degrees C, 27 degrees C and 37 degrees C whereas the amplitude approximately doubled with each 10 degree C rise in temperature (Q(10)congruent with 2). The periodicity was observed with both intact milk fat globule membranes and with detergent-solubilized membranes, demonstrating that the oscillations did not require an association with membranes. The periodicity was not the result of instrument variation or of chemical interactions among reactants in solution. Preparations with different periodicities entrained (autosynchronized) when mixed. Upon mixing, the preparations exhibited two oscillatory patterns but eventually a single pattern representing the mean of the farthest separated maxima of the two preparations analyzed separately emerged. The cell surface NOX protein is the first reported example of an entrainable biochemical entity with a temperature-compensated periodicity potentially capable of functioning as an ultradian or circadian clock driver.

  13. Comparative 2D-DIGE Proteomic Analysis of Bovine Mammary Epithelial Cells during Lactation Reveals Protein Signatures for Lactation Persistency and Milk Yield

    Science.gov (United States)

    Janjanam, Jagadeesh; Singh, Surender; Jena, Manoj K.; Varshney, Nishant; Kola, Srujana; Kumar, Sudarshan; Kaushik, Jai K.; Grover, Sunita; Dang, Ajay K.; Mukesh, Manishi; Prakash, B. S.; Mohanty, Ashok K.

    2014-01-01

    Mammary gland is made up of a branching network of ducts that end with alveoli which surrounds the lumen. These alveolar mammary epithelial cells (MEC) reflect the milk producing ability of farm animals. In this study, we have used 2D-DIGE and mass spectrometry to identify the protein changes in MEC during immediate early, peak and late stages of lactation and also compared differentially expressed proteins in MEC isolated from milk of high and low milk producing cows. We have identified 41 differentially expressed proteins during lactation stages and 22 proteins in high and low milk yielding cows. Bioinformatics analysis showed that a majority of the differentially expressed proteins are associated in metabolic process, catalytic and binding activity. The differentially expressed proteins were mapped to the available biological pathways and networks involved in lactation. The proteins up-regulated during late stage of lactation are associated with NF-κB stress induced signaling pathways and whereas Akt, PI3K and p38/MAPK signaling pathways are associated with high milk production mediated through insulin hormone signaling. PMID:25111801

  14. Bovine milk proteome in the first 9 days: protein interactions in maturation of the immune and digestive system of the newborn.

    Directory of Open Access Journals (Sweden)

    Lina Zhang

    Full Text Available In order to better understand the milk proteome and its changes from colostrum to mature milk, samples taken at seven time points in the first 9 days from 4 individual cows were analyzed using proteomic techniques. Both the similarity in changes from day 0 to day 9 in the quantitative milk proteome, and the differences in specific protein abundance, were observed among four cows. One third of the quantified proteins showed a significant decrease in concentration over the first 9 days after calving, especially in the immune proteins (as much as 40 fold. Three relative high abundant enzymes (XDH, LPL, and RNASE1 and cell division and proliferation protein (CREG1 may be involved in the maturation of the gastro-intestinal tract. In addition, high correlations between proteins involved in complement and blood coagulation cascades illustrates the complex nature of biological interrelationships between milk proteins. The linear decrease of protease inhibitors and proteins involved in innate and adaptive immune system implies a protective role for protease inhibitor against degradation. In conclusion, the results found in this study not only improve our understanding of the role of colostrum in both host defense and development of the newborn calf but also provides guidance for the improvement of infant formula through better understanding of the complex interactions between milk proteins.

  15. Role of xanthine oxidase, lactoperoxidase, and NO in the innate immune system of mammary secretion during active involution in dairy cows: manipulation with casein hydrolyzates.

    Science.gov (United States)

    Silanikove, Nissim; Shapiro, Fira; Shamay, Avi; Leitner, Gabriel

    2005-05-01

    The aims of this study were to test whether xanthine oxidase, lactoperoxidase, and NO are components of the innate immune system of mammary secretion during active involution in dairy cows, and whether the innate immune system is activated by casein hydrolysates. Our laboratory has shown recently that infusion of CNH into mammary glands induced involution and was associated with earlier increases in the concentrations of components of the innate immune system. Intact casein is inactive and served as control. Half of the glands of 8 Holstein cows scheduled for dry off (approximately 60 days before parturition) were injected for 3 days with a single dose of casein hydrolyzates and the contralateral glands with a single dose of intact casein with the same concentration. Involution elicited marked increases in xanthine oxidase and lactoperoxidase activities, and accumulation of urate and nitrate. NO and H(2)O(2) were constantly produced in the mammary gland secretion. Nitrite formed either by autooxidation of NO or by conversion of nitrate to nitrite by xanthine oxidase was converted into the powerful nitric dioxide radical by lactoperoxidase and H(2)O(2) that is derived from the metabolism of xanthine oxidase. Nitric dioxide is most likely responsible for the formation of nitrosothiols on thiol-bearing groups, which allows an extended NO presence in mammary secretion. Nitrite is effectively converted to nitrate, which accumulated in the range of approximately 25 microM -1 mM from the start of the experiment to the complete involution of glands. The mammary secretion in all glands was bactericidal and bacteriostatic during established involution, and this appeared sooner and more acutely in glands treated with casein hydrolyzates, within 8 to 24 h. It is concluded that xanthine oxidase, lactoperoxidase, and NO are components of the mammary innate immune system that form bactericidal and bacteriostatic activities in mammary secretions. The innate immune system play a

  16. Mastitis Bovina: Resistencia a antibióticos de cepas de Staphylococcus aureus asiladas de leche (Bovine Mastitis: Antimicrobial resistance of Staphylococcus aureus strains isolated from milk

    Directory of Open Access Journals (Sweden)

    Pellegrino, MS

    2011-07-01

    Full Text Available ResumenLa mastitis bovina es considerada la enfermedad infecciosa del ganado lechero de mayor impacto económico mundial, siendo Staphylococcus aureus el principal agente patógeno en muchos países.SummaryBovine mastitis is a frequent cause of economic loss in worldwide dairy herds, being Staphylococcus aureus the main etiological agent in many countries.

  17. Mastitis Bovina: Resistencia a antibióticos de cepas de Staphylococcus aureus asiladas de leche (Bovine Mastitis: Antimicrobial resistance of Staphylococcus aureus strains isolated from milk)

    OpenAIRE

    Pellegrino, MS; Frola, ID; Odierno, LM; Bogni, CI

    2011-01-01

    ResumenLa mastitis bovina es considerada la enfermedad infecciosa del ganado lechero de mayor impacto económico mundial, siendo Staphylococcus aureus el principal agente patógeno en muchos países.SummaryBovine mastitis is a frequent cause of economic loss in worldwide dairy herds, being Staphylococcus aureus the main etiological agent in many countries.

  18. Feeding bovine milks with low or high IgA levels is associated with altered re-establishment of murine intestinal microbiota after antibiotic treatment

    Science.gov (United States)

    Young, Wayne; Cakebread, Julie A.; Haigh, Brendan J.

    2016-01-01

    Antibiotics are a vital and commonly used therapeutic tool, but their use also results in profound changes in the intestinal microbiota that can, in turn, have significant health consequences. Understanding how the microbiota recovers after antibiotic treatment will help to devise strategies for mitigating the adverse effects of antibiotics. Using a mouse model, we have characterized the changes occurring in the intestinal microbiota immediately after five days exposure to ampicillin, and then at three and fourteen days thereafter. During the fourteen day period of antibiotic recovery, groups of mice were fed either water, cows’ milk containing high levels of IgA, or cows’ milk containing low levels of IgA as their sole source of liquid. Effects on microbiota of feeding milks for 14 days were also assessed in groups of mice that had no ampicillin exposure. Changes in microbiota were measured by high throughput sequencing of the V4 to V6 variable regions of the 16S ribosomal RNA gene. As expected, exposure to ampicillin led to profound changes to the types and abundance of bacteria present, along with a loss of diversity. At 14 days following antibiotic exposure, mice fed water had recovered microbiota compositions similar to that prior to antibiotics. However, feeding High-IgA milk to mice that has been exposed to antibiotics was associated with altered microbiota compositions, including increased relative abundance of Lactobacillus and Barnesiella compared to the start of the study. Mice exposed to antibiotics then fed Low-IgA milk also showed increased Barnesiella at day 14. Mice without antibiotic perturbation, showed no change in their microbiota after 14 days of milk feeding. Overall, these findings add to a knowledge platform for optimizing intestinal function after treatment with antibiotics in the human population. PMID:27703861

  19. Feeding bovine milks with low or high IgA levels is associated with altered re-establishment of murine intestinal microbiota after antibiotic treatment

    Directory of Open Access Journals (Sweden)

    Alison J. Hodgkinson

    2016-09-01

    Full Text Available Antibiotics are a vital and commonly used therapeutic tool, but their use also results in profound changes in the intestinal microbiota that can, in turn, have significant health consequences. Understanding how the microbiota recovers after antibiotic treatment will help to devise strategies for mitigating the adverse effects of antibiotics. Using a mouse model, we have characterized the changes occurring in the intestinal microbiota immediately after five days exposure to ampicillin, and then at three and fourteen days thereafter. During the fourteen day period of antibiotic recovery, groups of mice were fed either water, cows’ milk containing high levels of IgA, or cows’ milk containing low levels of IgA as their sole source of liquid. Effects on microbiota of feeding milks for 14 days were also assessed in groups of mice that had no ampicillin exposure. Changes in microbiota were measured by high throughput sequencing of the V4 to V6 variable regions of the 16S ribosomal RNA gene. As expected, exposure to ampicillin led to profound changes to the types and abundance of bacteria present, along with a loss of diversity. At 14 days following antibiotic exposure, mice fed water had recovered microbiota compositions similar to that prior to antibiotics. However, feeding High-IgA milk to mice that has been exposed to antibiotics was associated with altered microbiota compositions, including increased relative abundance of Lactobacillus and Barnesiella compared to the start of the study. Mice exposed to antibiotics then fed Low-IgA milk also showed increased Barnesiella at day 14. Mice without antibiotic perturbation, showed no change in their microbiota after 14 days of milk feeding. Overall, these findings add to a knowledge platform for optimizing intestinal function after treatment with antibiotics in the human population.

  20. A proteomic perspective on the changes in milk proteins due to high somatic cell count

    NARCIS (Netherlands)

    Zhang, L.; Boeren, J.A.; Hooijdonk, van A.C.M.; Vervoort, J.J.M.; Hettinga, K.A.

    2015-01-01

    Although cows with subclinical mastitis have no difference in the appearance of their milk, milk composition and milk quality are altered because of the inflammation. To know the changes in milk quality with different somatic cell count (SCC) levels, 5 pooled bovine milk samples with SCC from 105 to

  1. Caracterización de un proceso operativo de análisis de calidad higiénica y sanitaria de la leche bovina cruda, con equipos automatizados BactoScan FC 50 y CombiFoss FT+ 200, en un laboratorio de calidad de leche (Characterization of the operative process for the analysis the hygienic and sanitary quality the raw bovine milk, with automated equipment BactoScan FC 50 and CombiFoss FT+ 200, in a quality milk laboratory .

    Directory of Open Access Journals (Sweden)

    Gaona Yira

    2011-04-01

    Full Text Available ResumenLa caracterización del proceso operativo de análisis de calidad higiénica y sanitaria de la leche bovina cruda con los equipos automatizados BactoScanFC50 y CombiFossFT+200, se realizó de forma separada para las fase preanalítica, analítica y postanalítica del proceso, para cada una de ellas se presentan las condiciones y especificaciones requeridas en equipos, personal, documentación , procedimientos, infraestructura y ambiente de trabajo que permiten la transformación de las muestras de leche bovina cruda desde su ingreso al laboratorio de calidad de leche, hasta la emisión de un resultado final de calidad para el usuario solicitante del servicio de análisis de calidad higiénica y sanitaria de la leche.SummaryThe characterization the operative process analysis the hygienic and sanitary quality the raw bovine milk with the automated equipment BactoScanFC50 and CombiFossFT+200, was carried out separate form for the phase preanalytic, analytic and postanalytic the process, for each one them the conditions and specifications are presented equired in equipments, personal, documentation, procedures, infrastructure and atmosphere work that allow the transformation the samples raw bovine milk from its entrance to the laboratory quality milk, until the emission final result quality for the userapplicant the service analysis the hygienic and sanitary quality milk.

  2. The use of Fourier-transform infrared spectroscopy to predict cheese yield and nutrient recovery or whey loss traits from unprocessed bovine milk samples.

    Science.gov (United States)

    Ferragina, A; Cipolat-Gotet, C; Cecchinato, A; Bittante, G

    2013-01-01

    Cheese yield is an important technological trait in the dairy industry in many countries. The aim of this study was to evaluate the effectiveness of Fourier-transform infrared (FTIR) spectral analysis of fresh unprocessed milk samples for predicting cheese yield and nutrient recovery traits. A total of 1,264 model cheeses were obtained from 1,500-mL milk samples collected from individual Brown Swiss cows. Individual measurements of 7 new cheese yield-related traits were obtained from the laboratory cheese-making procedure, including the fresh cheese yield, total solid cheese yield, and the water retained in curd, all as a percentage of the processed milk, and nutrient recovery (fat, protein, total solids, and energy) in the curd as a percentage of the same nutrient contained in the milk. All individual milk samples were analyzed using a MilkoScan FT6000 over the spectral range from 5,000 to 900 wavenumber × cm(-1). Two spectral acquisitions were carried out for each sample and the results were averaged before data analysis. Different chemometric models were fitted and compared with the aim of improving the accuracy of the calibration equations for predicting these traits. The most accurate predictions were obtained for total solid cheese yield and fresh cheese yield, which exhibited coefficients of determination between the predicted and measured values in cross-validation (1-VR) of 0.95 and 0.83, respectively. A less favorable result was obtained for water retained in curd (1-VR=0.65). Promising results were obtained for recovered protein (1-VR=0.81), total solids (1-VR=0.86), and energy (1-VR=0.76), whereas recovered fat exhibited a low accuracy (1-VR=0.41). As FTIR spectroscopy is a rapid, cheap, high-throughput technique that is already used to collect standard milk recording data, these FTIR calibrations for cheese yield and nutrient recovery highlight additional potential applications of the technique in the dairy industry, especially for monitoring cheese

  3. Inhibitory activity of xanthine oxidase by fractions Crateva adansonii

    Directory of Open Access Journals (Sweden)

    A Abdullahi

    2012-01-01

    Conclusions: Enzyme inhibition mechanism indicated that the mode of inhibition was of a mixed type. Our findings suggest that the therapeutic use of these plants may be due to the observed Xanthine oxidase inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.

  4. Antioxidant effect of naturally occurring xanthines on the oxidative damage of DNA bases; Effet antioxydant de xanthines naturelles sur le dommage oxydant des bases de l`ADN

    Energy Technology Data Exchange (ETDEWEB)

    Vieira, A.J.S.C.; Telo, J.P.; Pereira, H.F.; Patrocinio, P.F. [Instituto Superior Tecnico, Lisbon (Portugal); Dias, R.M.B. [Instituto Tecnologico e Nuclear, Sacavem codex (Portugal). Dept. de Quimica

    1999-01-01

    The repair of the oxidised radicals of adenine and guanosine by several naturally occurring xanthines was studied. Each pair of DNA purine/xanthine was made to react with the sulphate radical and the decrease of the concentration of both compounds was measured by HPLC as a function of irradiation time. The results show that xanthine efficiently prevents the oxidation of the two DNA purines. Theophylline and para-xanthine repair the oxidizes radical of adenine but not the one from guanosine. Theobromine and caffeine to do not show any protecting effect. An order of the oxidation potentials of all the purines studied is proposed. (authors) 10 refs.

  5. Comments on "Detection of Toxoplasma gondii in raw caprine, ovine, bovine, and camel milk using cell cultivation, cat bioassay, capture ELISA, and PCR methods in Iran"

    Science.gov (United States)

    Toxoplasmosis is a parasitic infection of humans and animals and it continues to be public health and food safety issue. Among the many ways this infection is acquired, the consumption of unpasteurized goat cheese and milk has been suggested as a risk factor for toxoplasmosis in humans. Although pas...

  6. Efficiency of N2 gas flushing compared to the lactoperoxidase system at controlling bacterial growth in bovine raw milk stored at mild temperatures

    Directory of Open Access Journals (Sweden)

    Patricia eMunsch-Alatossava

    2016-06-01

    Full Text Available To prevent excessive bacterial growth in raw milk, the FAO recommends two options: either cold storage or activation of the lactoperoxidase system (LPs/ HT in milk with the addition of two chemical preservatives, hydrogen peroxide (H and thiocyanate (T. N2 gas flushing of raw milk has shown great potential to control bacterial growth in a temperature range of 6 to 12°C without promoting undesired side effects. Here, the effect of N2 gas (N was tested as a single treatment and in combination with the lactoperoxidase system (NHT on seven raw milk samples stored at 15 or 25°C. For the ratio defined as bacterial counts from a certain treatment/counts on the corresponding control, a classical Analyse of Variance (ANOVA was performed, followed by mean comparison with the Ryan-Einot-Gabriel-Welsch multiple range test (REGWQ. Altogether, the growth inhibition was slightly but significantly higher at 25°C than at 15°C. Except for one sample, all ratios were lower for HT than for N alone; however, these differences were not judged to be significant for five samples by the REGWQ test; in the remaining two samples, N was more effective than HT in one case and less effective in the other case. This study shows that N2 gas flushing, which inhibited bacterial growth in raw milk at 15 and 25°C for 24 and 12 h, respectively, could constitute an alternative to LPs where no cold storage facilities exist, especially as a replacement for adulterating substances.

  7. Efficiency of N2 Gas Flushing Compared to the Lactoperoxidase System at Controlling Bacterial Growth in Bovine Raw Milk Stored at Mild Temperatures

    Science.gov (United States)

    Munsch-Alatossava, Patricia; Quintyn, Romanie; De Man, Ingrid; Alatossava, Tapani; Gauchi, Jean-Pierrre

    2016-01-01

    To prevent excessive bacterial growth in raw milk, the FAO recommends two options: either cold storage or activation of the lactoperoxidase system (LPs/HT) in milk with the addition of two chemical preservatives, hydrogen peroxide (H) and thiocyanate (T). N2 gas flushing of raw milk has shown great potential to control bacterial growth in a temperature range of 6–12°C without promoting undesired side effects. Here, the effect of N2 gas (N) was tested as a single treatment and in combination with the lactoperoxidase system (NHT) on seven raw milk samples stored at 15 or 25°C. For the ratio defined as bacterial counts from a certain treatment/counts on the corresponding control, a classical Analyse of Variance (ANOVA) was performed, followed by mean comparison with the Ryan-Einot-Gabriel-Welsch multiple range test (REGWQ). Altogether, the growth inhibition was slightly but significantly higher at 25°C than at 15°C. Except for one sample, all ratios were lower for HT than for N alone; however, these differences were not judged to be significant for five samples by the REGWQ test; in the remaining two samples, N was more effective than HT in one case and less effective in the other case. This study shows that N2 gas flushing, which inhibited bacterial growth in raw milk at 15 and 25°C for 24 and 12 h, respectively, could constitute an alternative to LPs where no cold storage facilities exist, especially as a replacement for adulterating substances. PMID:27313575

  8. Human Milk Fortification.

    Science.gov (United States)

    Simmer, Karen

    2015-01-01

    Human milk is the feed of choice for preterm infants. However, human milk does not provide enough nutrition, especially protein, for preterm infants to achieve target growth rates similar to those in utero (15-20 g/kg per day). Fortifiers for human milk, manufactured from bovine milk, are commercially available and routinely used for patients born milk fortifier that is manufactured from donor human milk is available in some developed countries and may confer some clinical benefits, including a reduction in necrotizing enterocolitis. Fortification can be added in a standardized protocol as per manufacturers' instructions. Human milk composition can be analyzed and fortification individualized to take into account the large variation from mother to mother. Alternatively, fortification can be increased in a stepwise manner based on assumed composition while monitoring blood urea levels for safety. The current aim is to prevent preterm infants dropping percentiles and falling below the 10th percentile at 36 weeks' corrected gestational age or discharge home. More data are required on how best to fortify human milk for preterm infants to achieve optimal growth, development and health outcomes in the long term. There is an urgent need for well-designed and informed randomized clinical trials in this vulnerable preterm population.

  9. Proteins of human milk. I. Identification of major components

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, N.G.; Powers, M.T.; Tollaksen, S.L.

    1982-04-01

    Traditionally, human milk proteins are identified largely by reference to bovine milk. Hence, to identify the major proteins in human milk, we subjected human and bovine milk, in parallel, to high-resolution two-dimensional electrophoresis. Isoelectric precipitation at pH 4.6 was our criterion for distinguishing whey proteins from those of the casein complex. The ..cap alpha..- and..beta..-caseins were identified on the basis of relative abundance, relative molecular mass, and relative isoelectric points. No protein disappeared from ISO-DALT patterns of human milk after rennin treatment, and no new protein comparable to bovine para K-casein appeared in the BASO-DALT patterns; this suggests that K-casein is absent from human milk. The proteins identified in human milk patterns include the ..cap alpha.. and ..beta.. casein families, lactalbumin, albumin, transferrin, IgA, and lactoferrin. Numerous additional proteins seen in patterns for human milk remain to be identified.

  10. Human Milk Banking-Facts and Issues to Resolve

    OpenAIRE

    Willemijn E. Corpeleijn; Vermeulen, Marijn J.; Ineke van Vliet; I. Caroline Kruger; van Goudoever, Johannes B.

    2010-01-01

    The number of human milk banks is increasing worldwide. Although the beneficial effects of feeding premature infants with their mother’s milk are well documented, less is known about the effects of feeding these infants with pasteurized donor milk. We propose a randomized trial comparing the effects of a 100% human milk-based diet (human milk supplemented with a human milk-derived fortifier) and a diet (partially) based on bovine milk. In theory, human milk has a beneficial effect on various ...

  11. Mechanism of porcine liver xanthine oxidoreductase mediated N-oxide reduction of cyadox as revealed by docking and mutagenesis studies.

    Directory of Open Access Journals (Sweden)

    Chigang Chen

    Full Text Available Xanthine oxidoreductase (XOR is a cytoplasmic molybdenum-containing oxidoreductase, catalyzing both endogenous purines and exogenous compounds. It is suggested that XOR in porcine hepatocytes catalyzes the N-oxide reduction of quinoxaline 1,4-di-N-oxides (QdNOs. To elucidate the molecular mechanism underlying this metabolism, the cDNA of porcine XOR was cloned and heterologously expressed in Spodoptera frugiperda insect cells. The bovine XOR, showing sequence identity of 91% to porcine XOR, was employed as template for homology modeling. By docking cyadox, a representative compound of QdNOs, into porcine XOR model, eight amino acid residues, Gly47, Asn352, Ser360, Arg427, Asp430, Asp431, Ser1227 and Lys1230, were located at distances of less than 4Å to cyadox. Site-directed mutagenesis was performed to analyze their catalytic functions. Compared with wild type porcine XOR, G47A, S360P, D431A, S1227A, and K1230A displayed altered kinetic parameters in cyadox reduction, similarly to that in xanthine oxidation, indicating these mutations influenced electron-donating process of xanthine before subsequent electron transfer to cyadox to fulfill the N-oxide reduction. Differently, R427E and D430H, both located in the 424-434 loop, exhibited a much lower K(m and a decreased V(max respectively in cyadox reduction. Arg427 may be related to the substrate binding of porcine XOR to cyadox, and Asp430 is suggested to be involved in the transfer of electron to cyadox. This study initially reveals the possible catalytic mechanism of porcine XOR in cyadox metabolism, providing with novel insights into the structure-function relationship of XOR in the reduction of exogenous di-N-oxides.

  12. Treatment of Bovine Milk by Horseradish Peroxidase and Its Impact on the Quality of Set-yoghurt%牛乳的过氧化物酶处理与凝固型酸奶品质的变化

    Institute of Scientific and Technical Information of China (English)

    温研; 赵新淮

    2011-01-01

    利用辣根过氧化物酶和阿魏酸处理牛乳并制备凝固型酸奶,采用质构仪和流变仪分别对不同酸奶样品的质构和流变特性进行对比研究.研究结果表明,辣根过氧化物酶、阿魏酸和明胶的添加对酸奶样品的主要成分组成没有影响,但对酸奶质构和流变特性的影响较大.与对照样相比,添加明胶、添加酶和阿魏酸、添加酶酸奶样品的硬度分别增加38.03%、34.07%和17.08%,黏度分别增加45.13%、40.85%和28.36%,而乳清析出率分别降低12.02%、6.28%和3.41%.同时,酸奶的表观黏度、触变性和粘弹性也都增加,增幅最大者为添加明胶酸奶样品,其次是添加酶和阿魏酸的酸奶样品,增幅最小的是添加酶的酸奶样品.研究结果显示利用辣根过氧化物酶、阿魏酸处理牛乳可以改善凝固型酸奶的品质.%Bovine milk was treated with horseradish peroxidase or/and ferulic acid, and used to prepare set-yoghurt.The textural and rheological properties of the yoghurt samples prepared were measured with Texture Profile Analyzer and Rheometer and compared to that of control.The results showed that treatment of bovine milk with horseradish peroxidase or/and ferulic acid did not impact the main compositions of the yoghurt samples, but clear influence on the texture and rheology.Compared to that of control, the hardness or the adhesiveness of the yoghurt samples with gelatin addition, horseradish peroxidase treatment, or horseradish peroxidase plus ferulic acid treatment had an increase of 38.03, 34.07 and 17.08% , or 45.13 , 40.85 and 28.36% , respectively, while the syneresis of these samples had a decrease of 12.02, 6.28 and 3.41% , respectively.Also, the apparent viscosities, thixotropy and viscoelastic behaviors of these samples were in the order of the yoghurt with gelatin addition, the yoghurt with horseradish peroxidase plus ferulic acid treatment, the yoghurt samples with horseradish peroxidase treatment

  13. Genetic and environmental relationships of different measures of individual cheese yield and curd nutrients recovery with coagulation properties of bovine milk.

    Science.gov (United States)

    Cecchinato, A; Bittante, G

    2016-03-01

    The aim of this study was to elucidate the relationships between various cheesemaking-related traits, namely the well-known traditional milk coagulation properties (MCP), the new curd firming and syneresis traits, the cheese yield, and the curd nutrient recoveries or whey losses (all measured at the individual level). Data were obtained from 1,167 Brown Swiss cows reared in 85 herds. A 2-L milk sample was collected once from each animal and assessed for 10 phenotypes related to changes in curd firmness (CF) over time, plus 7 cheesemaking traits. The CF-related traits included 4 traditional single-point lactodynamographic properties [rennet coagulation time (RCT, min); time to a CF of 20mm, min; and the CF 30 and 45 min after rennet addition (a30 and a45, respectively)], 4 parameters used to model the 360 CF data recorded over time for each milk sample [the potential asymptotic CF at infinite time (CFP, mm); the CF instant rate constant, % × min(-1); the syneresis instant rate constant, % × min(-1); and the RCT obtained from modeling individual samples], and 2 traits calculated from individual equations [the maximum CF(CFmax, mm); and the time at CFmax, min]. The cheesemaking traits included 3 cheese yield traits (weights of the fresh curd, curd solids and curd moisture as percent of the weights of the processed milk) and 4 milk nutrient recoveries in the curd (calculated as the percent ratios between a given nutrient in the curd versus that in the processed milk). Bayesian methodology-based multivariate analyses were used to estimate the phenotypic, additive genetic, herd/date, and residual relationships between the aforementioned traits, whereas statistical inferences were based on the marginal posterior distributions of the parameters of concern. The a45, CFP, and CFmax traits were genetically associated with all of the percent cheese yield traits (the additive genetic correlations varied from 0.752 to 0.855 for a45; 0.496 to 0.583 for CFP; and 0.750 to 0

  14. Construction of transgenic mice producing CAT to milk by co-injection of two overlapping fragments of bovine αs1-casein-CAT gene

    Institute of Scientific and Technical Information of China (English)

    劳为德; 刘伟; 成国祥; 徐少甫; 成勇

    1996-01-01

    Two αs1-casein/chloramphenicol acetyltransferase (CAT) gene constructs overlapping by 3.0kb were constructed and co-injected into murine zygotes. In 9 of 10 lines of transgenic mice obtained, based on analysis of structure and expression of the transgene, accurate extrachromosomal homologous recombination (ECR) between the two overlapping DNA fragments was found. Different levels of CAT activity were detected in milk from these lines. The highest CAT activity was about 25-50μg/mL milk. In some mice. CAT activity was found in salvia gland, thymus and spleen extracts. The high frequency and accuracy of ECR reported here will be applicable in the experimental manipulation for generation of relatively large transgene.

  15. Xanthine Biosensor Based on Didodecyldimethylammonium Bromide Modified Pyrolytic Graphite Electrode

    Institute of Scientific and Technical Information of China (English)

    TANG,Ji-Lin(唐纪琳); HAN,Xiao-Jun(韩晓军); HUANG,Wei-Min(黄卫民); WANG,Er-Kang(汪尔康)

    2002-01-01

    The vesicle of didodecyldimethylammonium bromide (DDAB)which contained tetrathiafulvalene (TTF) was mixed with xanthine oxidase, and the mixture was cast on the pyrolytic graphite electrode. The lipid films were used to supply a biological environment resembling biomembrane on the surface of the electrode. TTF was used as a mediator because of its high electron-transfer efficiency. A novel xanthine biosensor based on cast DDAB film was developed. The effects of pH and operating potential were explored for optimum analytical performance by using the amperometric method. The response time of the biosensor was less than 10 s. The detection limit of the biosensor was 3.2 × 10-7 mol/L and the liner range was from 4 × 10-7 mol/L to 2.4 × 10-6 mol/L.

  16. Selective bronchodilators from 1-(5'-oxohexyl)xanthines.

    Science.gov (United States)

    Miyamoto, K I; Sakai, R; Yamamoto, Y; Konno, K; Sanae, F; Hasegawa, T; Takagi, K

    1992-11-01

    A series of twenty one 1-(5'-oxohexyl)xanthines substituted with alkyl chains at the N3 and N7 positions of the xanthine nucleus were prepared and their relaxant activity in guinea-pig isolated tracheal muscle and positive chronotropic activity in isolated right atrium of guinea-pig were compared. The tracheal relaxant activities were markedly increased with alkyl chain length at the N3 position, but decreased by the N7 alkylation. The positive chronotropic activities in the right atrium were increased by introduction of an n-propyl group at the N3 position but decreased by substitution of longer alkyl chains, and the action on the heart was diminished by N7 substitution. The activities of compounds on cAMP-phosphodiesterase (PDE) and binding of [3H]8-cyclopentyl-1,3-dipropylxanthine were measured in the homogenate of tracheal muscle and the membrane preparation of cerebral cortex, respectively. No relationship among tracheal muscle relaxant activity, cAMP-PDE inhibitory activity and adenosine antagonism of these xanthines was observed, and other action mechanisms should be considered for their relaxant activities. This study indicated that N3 alkylation is important for the selectivity for tracheal muscle, while the introduction of long alkyl chains such as n-butyl and n-pentyl groups at the N3 and N7 positions diminished the potency for the right atrium in guinea-pigs. 3-n-Pentyl- and 7-methyl-3-n-pentyl-1-(5'-oxohexyl)xanthines showed much higher bronchoselectivity than oxpentifylline and theophylline.

  17. CONDUCTIVITY OF DONKEY MILK

    Directory of Open Access Journals (Sweden)

    F. Conte

    2009-06-01

    Full Text Available The electrical conductivity (EC of milk is considered as one of the most important parameters which supports the diagnosis of mastitis in cows.Milk ions have a considerable influence on EC and their concentrations vary depending on animal species, season, lactation stage, etc. Some components of milk can change the EC, e.g. lactose. A negative correlation between EC values and the concentration of lactose is noticed, as a consequence of the inverse relation between this disaccharide and the chlorine content in milk. Fat and casein contents exert some influence on the EC, too. This study provides preliminary results on the physiological EC values in donkey milk and aims to highlight any correlation with some of its chemical-physical parameters and Somatic Cell Count (SCC. Mean EC value in donkey milk was found to be 3.57 mS. Statistically significant correlations were found between EC and SCC (r = 0.57 , p < 0.01 and between EC and (r = 0.30 , p < 0.05. The EC and lactose were not correlated although a reduction of EC was often observed when the lactose content increased, as reported in the literature for bovine milk. According to the EC can be considered as a reliable parameter to identify any breast disorder, taking into account the physiological factors that influence EC.

  18. Milk: the new sports drink? A Review.

    Science.gov (United States)

    Roy, Brian D

    2008-01-01

    There has been growing interest in the potential use of bovine milk as an exercise beverage, especially during recovery from resistance training and endurance sports. Based on the limited research, milk appears to be an effective post-resistance exercise beverage that results in favourable acute alterations in protein metabolism. Milk consumption acutely increases muscle protein synthesis, leading to an improved net muscle protein balance. Furthermore, when post-exercise milk consumption is combined with resistance training (12 weeks minimum), greater increases in muscle hypertrophy and lean mass have been observed. Although research with milk is limited, there is some evidence to suggest that milk may be an effective post-exercise beverage for endurance activities. Low-fat milk has been shown to be as effective, if not more effective, than commercially available sports drinks as a rehydration beverage. Milk represents a more nutrient dense beverage choice for individuals who partake in strength and endurance activities, compared to traditional sports drinks. Bovine low-fat fluid milk is a safe and effective post exercise beverage for most individuals, except for those who are lactose intolerant. Further research is warranted to better delineate the possible applications and efficacy of bovine milk in the field of sports nutrition.

  19. Milk: the new sports drink? A Review

    Directory of Open Access Journals (Sweden)

    Roy Brian D

    2008-10-01

    Full Text Available Abstract There has been growing interest in the potential use of bovine milk as an exercise beverage, especially during recovery from resistance training and endurance sports. Based on the limited research, milk appears to be an effective post-resistance exercise beverage that results in favourable acute alterations in protein metabolism. Milk consumption acutely increases muscle protein synthesis, leading to an improved net muscle protein balance. Furthermore, when post-exercise milk consumption is combined with resistance training (12 weeks minimum, greater increases in muscle hypertrophy and lean mass have been observed. Although research with milk is limited, there is some evidence to suggest that milk may be an effective post-exercise beverage for endurance activities. Low-fat milk has been shown to be as effective, if not more effective, than commercially available sports drinks as a rehydration beverage. Milk represents a more nutrient dense beverage choice for individuals who partake in strength and endurance activities, compared to traditional sports drinks. Bovine low-fat fluid milk is a safe and effective post exercise beverage for most individuals, except for those who are lactose intolerant. Further research is warranted to better delineate the possible applications and efficacy of bovine milk in the field of sports nutrition.

  20. Inhibition of Xanthine Oxidase Activity by Gnaphalium Affine Extract

    Institute of Scientific and Technical Information of China (English)

    Wei-qing Lin; Jian-xiang Xie; Xiao-mu Wu; Lin Yang; Hai-dong Wang

    2014-01-01

    Objective To evaluate the inhibitory effect of Gnaphalium affine extracts on xanthine oxidase (XO) activity in vitro and to analyze the mechanism of this effect. Methods In this in vitro study, Kinetic measurements were performed in 4 different inhibitor concentrations and 5 different xanthine concentrations (60, 100, 200, 300, 400 μmol/L). Dixon and Lineweaver-Burk plot analysis were used to determine Ki values and the inhibition mode for the compounds isolated from Gnaphalium affine extract. Results Four potent xanthine oxidase inhibitors were found in 95% ethanolic (v/v) Gnaphalium affine extract. Among them, the flavone Eupatilin exhibited the strongest inhibitory effect on XO with a inhibition constant (Ki) of 0.37μmol/L, lower than the Ki of allopurinol (4.56 mol/L), a known synthetic XO inhibitor. Apigenin (Ki of 0.56μmol/L, a proportion of 0.0053‰in Gnaphalium affine), luteolin (Ki of 2.63 μmol/L, 0.0032‰ in Gnaphalium affine) and 5-hydroxy-6,7,3’,4’-tetramethoxyflavone (Ki of 3.15μmol/L, 0.0043‰ in Gnaphalium affine) also contributed to the inhibitory effect of Gnaphalium affine extract on XO activity. Conclusions These results suggest that the use of Gnaphalium affine in the treatment of gout could be attributed to its inhibitory effect on XO. This study provides a rational basis for the traditional use of Gnaphalium affine against gout.

  1. Effects of alkyl substituents of xanthine on phosphodiesterase isoenzymes.

    Science.gov (United States)

    Miyamoto, K; Sakai, R; Kurita, M; Ohmae, S; Sanae, F; Sawanishi, H; Hasegawa, T; Takagi, K

    1995-03-01

    The structure-activity relationships of a series of alkylxanthine derivatives were investigated. The partition coefficient of alkylxanthines enlarged with an elongation of the alkyl chain at the 1-, 3-, or 7-position of xanthine. There was a mild correlation between the apparent partition coefficient and the tracheal relaxant activity or the inhibitory activity on phosphodiesterase (PDE) IV isoenzyme, while the tracheal relaxant activity closely correlated with the PDE IV inhibitory activity. Regarding substituents at different positions, the alkylation at the 3-position increased the inhibitory activity on every PDE isoenzyme. The alkylation at the 1-position potentiated the inhibitory activity on PDE IV with the alkyl chain length, but decreased the activities on other PDE isoenzymes. The alkylation at the 7-position was characteristic in its decrease in inhibitory activity on PDE III. These results suggested that the potency of the inhibitory activity of xanthine derivatives on PDE isoenzymes is not dependent simply upon their hydrophobicity but upon change in the affinity for the active sites on PDE isoenzymes by the introduction of the alkyl group at particular positions of the xanthine skeleton.

  2. Effects of alkyl substitutions of xanthine skeleton on bronchodilation.

    Science.gov (United States)

    Sakai, R; Konno, K; Yamamoto, Y; Sanae, F; Takagi, K; Hasegawa, T; Iwasaki, N; Kakiuchi, M; Kato, H; Miyamoto, K

    1992-10-30

    Structure-activity relationships in a series of 1,3,7-trialkyl-xanthine were studied with guinea pigs. Relaxant actions in the tracheal muscle were increased with alkyl chain length at the 1- and 3-positions of the xanthine skeleton, but decreased by alkylation at the 7-position. Positive chronotropic actions in the right atrium were potentiated with 3-alkyl chain length but tended to decrease with 1-alkylation and diminish by 7-substitution. Consequently, while the 1- and 3-substitutions were equally important for the tracheal smooth muscle relaxation, the substitution at the 1-position was more important than the 3-substitution for bronchoselectivity. The 7-alkylation may be significant to cancel heart stimulation. There were good correlations between the smooth muscle relaxant action and the cyclic AMP-PDE inhibitory activity in 3-substituents and the affinity for adenosine (A1) receptors in 1-, 3-, and 7-substituents. This suggests that not only the cyclic AMP-PDE inhibitory activity but also the adenosine antagonistic activity is important in the bronchodilatory effects of alkylxanthines. Among these xanthine derivatives, 1-butyl-3-propylxanthine and its 7-methylated derivative showed high bronchoselectivity in the in vitro and in vivo experiments compared to theophylline and enprofylline and may be new candidates for bronchodilator.

  3. Xanthine Oxidoreductase-Derived Reactive Species: Physiological and Pathological Effects.

    Science.gov (United States)

    Battelli, Maria Giulia; Polito, Letizia; Bortolotti, Massimo; Bolognesi, Andrea

    2016-01-01

    Xanthine oxidoreductase (XOR) is the enzyme that catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid and is widely distributed among species. In addition to this housekeeping function, mammalian XOR is a physiological source of superoxide ion, hydrogen peroxide, and nitric oxide, which can function as second messengers in the activation of various pathways. This review intends to address the physiological and pathological roles of XOR-derived oxidant molecules. The cytocidal action of XOR products has been claimed in relation to tissue damage, in particular damage induced by hypoxia and ischemia. Attempts to exploit this activity to eliminate unwanted cells via the construction of conjugates have also been reported. Moreover, different aspects of XOR activity related to phlogosis, endothelial activation, leukocyte activation, and vascular tone regulation, have been taken into consideration. Finally, the positive and negative outcomes concerning cancer pathology have been analyzed because XOR products may induce mutagenesis, cell proliferation, and tumor progression, but they are also associated with apoptosis and cell differentiation. In conclusion, XOR activity generates free radicals and other oxidant reactive species that may result in either harmful or beneficial outcomes.

  4. Xanthine Oxidoreductase-Derived Reactive Species: Physiological and Pathological Effects

    Directory of Open Access Journals (Sweden)

    Maria Giulia Battelli

    2016-01-01

    Full Text Available Xanthine oxidoreductase (XOR is the enzyme that catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid and is widely distributed among species. In addition to this housekeeping function, mammalian XOR is a physiological source of superoxide ion, hydrogen peroxide, and nitric oxide, which can function as second messengers in the activation of various pathways. This review intends to address the physiological and pathological roles of XOR-derived oxidant molecules. The cytocidal action of XOR products has been claimed in relation to tissue damage, in particular damage induced by hypoxia and ischemia. Attempts to exploit this activity to eliminate unwanted cells via the construction of conjugates have also been reported. Moreover, different aspects of XOR activity related to phlogosis, endothelial activation, leukocyte activation, and vascular tone regulation, have been taken into consideration. Finally, the positive and negative outcomes concerning cancer pathology have been analyzed because XOR products may induce mutagenesis, cell proliferation, and tumor progression, but they are also associated with apoptosis and cell differentiation. In conclusion, XOR activity generates free radicals and other oxidant reactive species that may result in either harmful or beneficial outcomes.

  5. Development of Fourier transform mid-infrared calibrations to predict acetone, β-hydroxybutyrate, and citrate contents in bovine milk through a European dairy network.

    Science.gov (United States)

    Grelet, C; Bastin, C; Gelé, M; Davière, J-B; Johan, M; Werner, A; Reding, R; Fernandez Pierna, J A; Colinet, F G; Dardenne, P; Gengler, N; Soyeurt, H; Dehareng, F

    2016-06-01

    To manage negative energy balance and ketosis in dairy farms, rapid and cost-effective detection is needed. Among the milk biomarkers that could be useful for this purpose, acetone and β-hydroxybutyrate (BHB) have been proved as molecules of interest regarding ketosis and citrate was recently identified as an early indicator of negative energy balance. Because Fourier transform mid-infrared spectrometry can provide rapid and cost-effective predictions of milk composition, the objective of this study was to evaluate the ability of this technology to predict these biomarkers in milk. Milk samples were collected in commercial and experimental farms in Luxembourg, France, and Germany. Acetone, BHB, and citrate contents were determined by flow injection analysis. Milk mid-infrared spectra were recorded and standardized for all samples. After edits, a total of 548 samples were used in the calibration and validation data sets for acetone, 558 for BHB, and 506 for citrate. Acetone content ranged from 0.020 to 3.355mmol/L with an average of 0.103mmol/L; BHB content ranged from 0.045 to 1.596mmol/L with an average of 0.215mmol/L; and citrate content ranged from 3.88 to 16.12mmol/L with an average of 9.04mmol/L. Acetone and BHB contents were log-transformed and a part of the samples with low values was randomly excluded to approach a normal distribution. The 3 edited data sets were then randomly divided into a calibration data set (3/4 of the samples) and a validation data set (1/4 of the samples). Prediction equations were developed using partial least square regression. The coefficient of determination (R(2)) of cross-validation was 0.73 for acetone, 0.71 for BHB, and 0.90 for citrate with root mean square error of 0.248, 0.109, and 0.70mmol/L, respectively. Finally, the external validation was performed and R(2) obtained were 0.67 for acetone, 0.63 for BHB, and 0.86 for citrate, with respective root mean square error of validation of 0.196, 0.083, and 0.76mmol/L. Although

  6. Multiresidue analysis of cephalosporin antibiotics in bovine milk based on molecularly imprinted polymer extraction followed by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Baeza, A N; Urraca, J L; Chamorro, R; Orellana, G; Castellari, M; Moreno-Bondi, M C

    2016-11-25

    This work reports the preparation of molecularly imprinted polymers (MIPs) selective to cephalosporin (CF) antibiotics, and their application as molecularly imprinted solid-phase extraction (MISPE) sorbents for the determination of these antimicrobials in milk samples. Several functional monomers and cross-linkers have been screened to select the best combination that provides high selectivity for the simultaneous multiresidue extraction of cefthiofur (THIO), cefazolin (AZO), cefquinome (QUI), cephapirin (API), cephalexin (ALE) and cephalonium (ALO) from the samples. The novel MIPs were prepared by a non-covalent imprinting approach in the form of spherical microparticles using the synthetic surrogate molecule sodium 7-(2-biphenylylcarboxamido)-3-methyl-3-cepheme-4-carboxylate, N-3,5-bis(trifluoromethyl)phenyl-N'-4-vinylphenyl urea (VPU) as functional monomer, and divinylbenzene (DVB) as crosslinking agent in a 1:2:20 molar ratio. The optimized MISPE method allowed the extraction of the target antimicrobials from raw cow milk samples using a selective washing with 5mL methanol/2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (HEPES) buffer (0.1M, pH 7.5) (2:98, v/v) to remove the non-specifically retained compounds, followed by elution with 1mL of trifluoroacetic acid (TFA) in methanol (0.1:99.9, v/v). The extracts have been analysed by UHPLC-MS/MS and the analytical method has been validated according to EU guideline 2002/657/EC. The limits of quantification (S/N=10) were in the 1.7-12.5μgkg(-1) range, well below the maximum residue limits (MRLs) currently established for the quantified cephalosporins in milk samples. The developed MIP allows mutiresidual determination of the six cephalosporin antibiotics mentioned above, significantly broadening the application to food analysis of MISPE methods.

  7. The reductive half-reaction of xanthine dehydrogenase from Rhodobacter capsulatus: the role of Glu232 in catalysis.

    Science.gov (United States)

    Hall, James; Reschke, Stefan; Cao, Hongnan; Leimkühler, Silke; Hille, Russ

    2014-11-14

    The kinetic properties of an E232Q variant of the xanthine dehydrogenase from Rhodobacter capsulatus have been examined to ascertain whether Glu(232) in wild-type enzyme is protonated or unprotonated in the course of catalysis at neutral pH. We find that kred, the limiting rate constant for reduction at high [xanthine], is significantly compromised in the variant, a result that is inconsistent with Glu(232) being neutral in the active site of the wild-type enzyme. A comparison of the pH dependence of both kred and kred/Kd from reductive half-reaction experiments between wild-type and enzyme and the E232Q variant suggests that the ionized Glu(232) of wild-type enzyme plays an important role in catalysis by discriminating against the monoanionic form of substrate, effectively increasing the pKa of substrate by two pH units and ensuring that at physiological pH the neutral form of substrate predominates in the Michaelis complex. A kinetic isotope study of the wild-type R. capsulatus enzyme indicates that, as previously determined for the bovine and chicken enzymes, product release is principally rate-limiting in catalysis. The disparity in rate constants for the chemical step of the reaction and product release, however, is not as great in the bacterial enzyme as compared with the vertebrate forms. The results indicate that the bacterial and bovine enzymes catalyze the chemical step of the reaction to the same degree and that the faster turnover observed with the bacterial enzyme is due to a faster rate constant for product release than is seen with the vertebrate enzyme.

  8. 反相高效液相色谱法测定牛奶中的主要蛋白质%Determination of Major Bovine Milk Proteins by Reversed Phase High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    王娟; 张庆合; 王志华; 李红梅

    2009-01-01

    A RP-HPLC method for the quantification of the six major bovine milk proteins (κ-casein (CN),α_(s2)-CN,α_(s1)-CN,β-CN,Whey,immunoglobulins (Igg) ) is described. Separation and quantification were achieved by a reversed phase analytical column (Agilent Zorbax 300SB-C_8,250 mm×4.6 mm ,5μm) and the gradient elution solvents of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in acetonitrile at a flow rate of 0. 8 mL/min. Column temperature was set at 45℃ and the sample was monitored with photodiode array detector at 214 nm. A linear relationship( r >0. 999) between the concentrations of proteins and peak areas was observed over the concentration range. Recoveries of six target proteins spiked in milk were form 74. 8% to 132.5%. Nine kinds of milks of different brands were analyzed,and the difference of the concentration and relative ration of κ-CN,α_(s2)-CN,α_(s1)-CN,β-CN and Whey were found.%建立了测定牛奶中的主要蛋白质的反相高效液相色谱法,对前处理方法进行了优化,采用Agilent Zorbax 300SB-C_8(250 mm×4.6 mm,5 μm)色谱柱,三氟乙酸-水-乙腈流动相梯度洗脱,214 nm检测,柱温45 ℃,外标法定量.测定κ-CN,α_(s2)-CN,α_(s1)-CN,β-CN,Whey和Igg的线性关系良好,相关系数均大于0 999,加标回收率在74.8%~132.5%之间.大豆蛋白质不影响分离与检测.采用本方法分析了9种牛奶样品中上述蛋白质的含量与总量,结果表明,本方法测定结果准确可靠.不同样品中4种酪蛋白与乳清蛋白的比例基本接近,但是不同品牌之间存在一定差异.

  9. Ocorrência de mastite bovina em fazendas produtoras de leite B no estado de São Paulo Occurrence of bovine mastitis in grade B milk farms in the state of São Paulo

    Directory of Open Access Journals (Sweden)

    L.F. Laranja

    1994-12-01

    Full Text Available Durante o período compreendido entre os meses de Março/1991 e Fevereiro /1992 foi realizado um experimento com o objetivo de avaliar a prevalência da mastite bovina em 07 fazendas produtoras de leite tipo B de diferentes regiões do estado de São Paulo. Foram analisadas 1683 vacas que deram origem a 7695 resultados do CMT e das quais foram coletadas 983 amostras de leite para realização do exame microbiológico. A análise do CMT indicou 47,0% de vacas com CMT negativo, 15,1% CMT + e 37,5% CMT ++/+ + + . Os escores do CMT foram distribuidos segundo o número de lactação (1ª lactação, 2ª lactação e 3ª ou + lactações e segundo o estágio de lactação (1-30 dias de lactação, 31-90 dias, 91-250 dias e > 250 dias. A análise dos dados demonstrou que houve um efeito significativo (p From March/1991 to February/1992 1,683 cows from seven grade "B" milk producing dairy farms were used in a study on the prevalence of bovine mastitis. GMT tests (7,695 were 47.0% negative; 15.1% " +" and 37.5% "++/+ + +". CMT scores were grouped by number of lactation (1st, 2nd, 3rd or more and by stage of lactation (1-30 days in milk; 31-90 days; 91-250 and > 250 days. Both stage of lactation and number of lactation had significant effect on % of cows CMT "++/+ + + ". Microbiological results from 983 milk samples were 50.97% positive. From the positive samples were isolated Staphylococcus sp (44.6%, Corynebacteríum sp (15.0%, Streptococcus sp (8.2%, Yeast/Fungi (5.4%. Bacillus sp (4.4%, E. coli (3.2% and Klebsiella sp (0.2%. Considering both pure and mixed culture isolation, Staphylococcus sp represents 55.6%, Corynebacteríum sp, 18.6% and Streptococcus sp, 15.0%.

  10. Effects of breed and casein genetic variants on protein profile in milk from Swedish Red, Danish Holstein, and Danish Jersey

    DEFF Research Database (Denmark)

    Gustavsson, Frida; Buitenhuis, Albert Johannes; Johansson, M

    2014-01-01

    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 techn......, for example, the processing of cheese...

  11. Genetic and biochemical evidence that recombinant Enterococcus spp. strains expressing gelatinase (GelE) produce bovine milk-derived hydrolysates with high angiotensin converting enzyme-inhibitory activity (ACE-IA).

    Science.gov (United States)

    Gútiez, Loreto; Borrero, Juan; Jiménez, Juan J; Gómez-Sala, Beatriz; Recio, Isidra; Cintas, Luis M; Herranz, Carmen; Hernández, Pablo E

    2014-06-18

    In this work, genes encoding gelatinase (gelE) and serine proteinase (sprE), two extracellular proteases produced by Enterococcus faecalis DBH18, were cloned in the protein expression vector pMG36c, containing the constitutive P32 promoter, generating the recombinant plasmids pCG, pCSP, and pCGSP encoding gelE, sprE, and gelE-sprE, respectively. Transformation of noncaseinolytic E. faecalis P36, E. faecalis JH2-2, E. faecium AR24, and E. hirae AR14 strains with these plasmids permitted detection of caseinolytic activity only in the strains transformed with pCG or pCGSP. Complementation of a deletion (knockout) mutant of E. faecalis V583 for production of gelatinase (GelE) with pCG unequivocally supported that gelE is responsible for the caseinolytic activity of the transformed strain grown in bovine skim milk (BSM). RP-HPLC-MS/MS analysis of hydrolysates of transformed Enterococcus spp. strains grown in BSM permitted the identification of 38 major peptide fragments including peptides with previously reported angiotensin converting enzyme-inhibitory activity (ACE-IA), antihypertensive activity, and antioxidant activity.

  12. Genetic variation in genes affecting milk composition and quality

    DEFF Research Database (Denmark)

    Bertelsen, Henriette Pasgaard

    In the past decade major advances in next generation sequencing technologies have provided new opportuneties for the detection of genetic variation. Combining the knowlegde of genetic variation with phenotypic distributions provides considerable possibilites for detection of candidate genes....... In addition, exploring genetic variation related to the major milk proteins of bovine milk indntified genetic variations with possitive effects on milk coagulation...

  13. Presença de resíduos de antibióticos em leite bovino pasteurizado Presence of antimicrobial residues in commercialized pasteurized bovine milk

    Directory of Open Access Journals (Sweden)

    Christina Maria Queiroz de Jesus Morais

    2010-05-01

    Full Text Available O objetivo deste trabalho foi avaliar a presença de resíduos de antibióticos no leite bovino pasteurizado comercializado na cidade do Rio de Janeiro e propor um programa de monitoramento à Vigilância Sanitária Municipal com base nesses dados. Durante o período de abril a agosto de 2006 foram analisadas 57 amostras de leite através de ensaios imunoenzimáticos para análise qualitativa dos betalactâmicos, tetraciclinas e estreptomicina/diidroestreptomicina na forma de kits. Os resultados evidenciaram que os antimicrobianos continuam sendo utilizados no tratamento do gado leiteiro.The aim of this work was to evaluate the antimicrobial residues present in pasteurized milk commercialized in the city of Rio de Janeiro Brazil and to suggest a monitoring program based on these data. During the period from April to August 2006, 57 milk samples were analyzed through and enzyme immunoassays for the qualitative analysis of beta-lactams, tetracyclines, and streptomycin/dihydrostreptomycin test Kits. The results showed that the studied antibiotics are have been used for the treatment of the dairy cattle.

  14. 响应面法优化牛乳蛋白的水解工艺研究%Response Surface Methodology Optimization of Bovine Milk Protein Hydrolysis

    Institute of Scientific and Technical Information of China (English)

    付莉; 岳喜庆; 赵凯

    2012-01-01

    为降低牛乳蛋白中的致敏蛋白含量,制备婴儿配方乳.利用蛋白酶水解牛乳蛋白,在筛选出最佳酶解用酶的基础上,以胰蛋白酶浓度、酶解时间、酶解温度为影响因子,在单因素试验结果的基础上,应用Centure-Composite Design中心组合方法进行三因素三水平的实验设计,以牛乳蛋白水解度为响应值,运用响应面法对水解条件进行进一步优化.结果表明:酶浓度、酶解时间对牛乳蛋白水解度影响显著;牛乳蛋白质水解的最佳工艺为:以胰蛋白酶为水解用酶,水解温度为54.65℃,酶添加量为0.79%,酶解时间为47.01 min.回归方程预测牛乳蛋白水解度理论值可达到20.78%;3次验证实验的平均水解度20.74%,与预测值相对误差为0.2%.说明利用响应面实验设计研究出胰蛋白酶水解牛乳蛋白最佳工艺,为婴儿配方乳的生产奠定了基础.%The aim was to reduce allergenic protein content in milk proteins for producing infant formula milk. We hydrolyzed milk protein based on screened best enzyme and based on trypsin concentration, hydrolysis time, hydrolysis temperature viewed as influence factors. We used single factor experiments and Centure-composite Design Center combination method of three factors and three levels experimental design to further optimize hydrolysis condition. The effect of enzyme concentration and hydrolysis time were significant. The optimum hydrolysis conditions were optimized as following: enzyme concentration 0.79%, hydrolysis temperature 54.65℃ and time 47.01 min. Regression equations predicting milk protein hydrolysis degree theory value could reached 20.78%, the average degree of hydrolysis of three times experiment could reached 20.74%, compared with predictive value the relative error was 0.2%. We successfully developed the optimum hydrolysis conditions of trypsin by response surface experimental design which laid a foundation on infant formula milk production.

  15. 牛乳蛋白抗原性研究现状概述%Research on antigenic proteins of bovine milk

    Institute of Scientific and Technical Information of China (English)

    宋宏新; 潘洁; 薛海燕

    2011-01-01

    Milk protein is an important indicator of the nutritional value of dairy products. The paper summarizes the major component, conformation,characteristic and the antigenicity of cow's protein. Specificity, interaction and the detection limitation of different determination were compared. In addition, the advantage and disadvantage of,enzyme linked immunosorbent assay were discussed.%乳蛋白是衡量乳制品营养价值的重要指标.文中介绍了牛乳蛋白的种类、结构、性质和抗原性强弱,比较了不同蛋白抗原的免疫抗体的特异性、交叉性、采用不同检测方法的检测限等.对酶联免疫吸附测定乳蛋白的优缺点进行了阐述,并展望了其应用前景.

  16. Potential estimation of titratable acidity in cow milk using mid-infrared spectrometry

    OpenAIRE

    Colinet, Frédéric; Soyeurt, Hélène; Anceau, Christine; Vanlierde, Amélie; Keyen, Nicolas; Pierre DARDENNE; Gengler, Nicolas; Sindic, Marianne

    2010-01-01

    Milk coagulation has a direct effect on cheese yield. Several factors influence the milk coagulation kinetics. In addition to calcium and milk protein concentrations, titratable acidity influences all the phases of milk coagulation. The objective of this research was to study the feasibility of prediction of titratable acidity directly in bovine milk using mid-infrared spectrometry. In order to maximize the variability in the measurements of titratable acidity, milk samples were collected on ...

  17. Subunit and whole molecule specificity of the anti-bovine casein immune response in recent onset psychosis and schizophrenia

    NARCIS (Netherlands)

    Severance, E.G.; Dickerson, F.B.; Halling, M.; Krivogorsky, B.; Haile, L.; Yang, S.; Stallings, C.R.; Origoni, A.E.; Bossis, I.; Xiao, J.; Dupont, D.; Haasnoot, W.; Yolken, R.H.

    2010-01-01

    Previous studies show increased antibody levels to bovine casein in some individuals with schizophrenia. The immunogenicity of specific domains of bovine casein varies among people with milk sensitivities and thus could vary among different neuropsychiatric disorders. Using ELISAs and immunoblotting

  18. [Forms of xanthine oxidoreductase in the tissues of Japanese quail].

    Science.gov (United States)

    Jankela, J; Baranovská, M; Antalíková, J

    1993-01-01

    The Japanese quail tissues--liver, kidney and pancreas were analysed for the presence of forms of xanthine oxidoreductase utilised cofactors NAD+, molecular oxygen or artificial acceptor--methylene blue, as well as for the validity of correlation between enzymatic activity and diet protein content. Four groups of animals with the experimental diets, the formulae of which are given in Tab. I, and control group with a commercial mash were fed for ten days. For enzyme preparation, the rough purification of cytoplasmic fraction with subsequent dialysis was used. The xanthine oxidoreductase utilised NAD+ (XOR-NAD) was detected in all examined tissues (Fig. 1), whereby the correlation of enzymatic activity with diet protein content was shown only in liver, according our previous findings (Jankela; 1978; Baranovská and Gazo, 1990). The values in liver and pancreas of animals fed a commercial mash were somewhat out of the range of linearity, probably because of the presence of nonprotein substances in mash, which affected the XOR activity in these organs (Jankela, 1992). The XOR utilised O2 (Fig. 2) was only detected in liver and kidney with certain activity in animals fed free protein diet. The percentage of this enzyme form was below 18% of the total activity (Fig. 5). The xanthine oxidoreductase utilised methylene blue (XOR-MM) was detected in liver, kidney and pancreas (Fig. 3). The correlation of enzymatic activity with diet protein content was linear in liver and kidney. The percentage of XOR-MM activity was very high, it amounted to 55% of the total activity (Fig. 4).(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Bioactive compounds of inhibiting xanthine oxidase from Selaginella labordei.

    Science.gov (United States)

    Tan, Wen-Jie; Xu, Jia-Cheng; Li, Li; Chen, Ke-Li

    2009-01-01

    Four flavone compounds were isolated from the effective fractions inhibiting xanthine oxidase (XOD) of the medicinal plant Selaginella labordei with anti-virus activity, and the structures were elucidated as 4'-methylether robustaflavone (1), robustaflavone (2), eriodictyol (3) and amentoflavone (4). The 50% inhibitory concentration (IC(50)) of the three compounds of inhibiting XOD were 61.0, 0.199, 16.0 and 32.0 mg L(-1), respectively. All of these compounds were isolated from the species for the first time, and eriodictyol was found from Selaginellaceae for the first time. Among these compounds, robustaflavone has been reported as an effective compound against the hepatitis B virus.

  20. Hydroxychavicol: a potent xanthine oxidase inhibitor obtained from the leaves of betel, Piper betle.

    Science.gov (United States)

    Murata, Kazuya; Nakao, Kikuyo; Hirata, Noriko; Namba, Kensuke; Nomi, Takao; Kitamura, Yoshihisa; Moriyama, Kenzo; Shintani, Takahiro; Iinuma, Munekazu; Matsuda, Hideaki

    2009-07-01

    The screening of Piperaceous plants for xanthine oxidase inhibitory activity revealed that the extract of the leaves of Piper betle possesses potent activity. Activity-guided purification led us to obtain hydroxychavicol as an active principle. Hydroxychavicol is a more potent xanthine oxidase inhibitor than allopurinol, which is clinically used for the treatment of hyperuricemia.

  1. Structure and Function of Bovine and Camel Chymosin

    DEFF Research Database (Denmark)

    Jensen, Jesper Langholm

    The central step in cheese making is the separation of milk into curd and whey. This can be done enzymatically by hydrolysis of the Phe105-Met106 bond or nearby bonds in bovine κ-casein, which releases its hydrophilic C-terminal leading to coagulation of the milk. The preferred enzyme...... for this action is bovine chymosin from the calf's stomach, as it has a high activity towards the Phe105-Met106 bond, and a low activity towards other bonds in the milk proteins, as the latter can lead to loss of protein in the whey and release of peptides with a bitter taste. Chymosin was isolated from camel...... this difference through the study of the structures of bovine and camel chymosin, and preparation of catalytically inactive enzymes in complex with substrate. Their milk-clotting activities was determined using the traditional assay on skimmed milk, and a fluorescence resonance energy transfer (FRET) assay...

  2. Proteomics of the milk fat globule membrane from Camelus dromedarius.

    Science.gov (United States)

    Saadaoui, Besma; Henry, Céline; Khorchani, Touhami; Mars, Mohamed; Martin, Patrice; Cebo, Christelle

    2013-04-01

    Camel milk has been widely characterized with regards to casein and whey proteins. However, in camelids, almost nothing is known about the milk fat globule membrane (MFGM), the membrane surrounding fat globules in milk. The purpose of this study was thus to identify MFGM proteins from Camelus dromedarius milk. Major MFGM proteins (namely, fatty acid synthase, xanthine oxidase, butyrophilin, lactadherin, and adipophilin) already evidenced in cow milk were identified in camel milk using MS. In addition, a 1D-LC-MS/MS approach led us to identify 322 functional groups of proteins associated with the camel MFGM. Dromedary MFGM proteins were then classified into functional categories using DAVID (the Database for Annotation, Visualization, and Integrated Discovery) bioinformatics resources. More than 50% of MFGM proteins from camel milk were found to be integral membrane proteins (mostly belonging to the plasma membrane), or proteins associated to the membrane. Enriched GO terms associated with MFGM proteins from camel milk were protein transport (p-value = 1.73 × 10(-14)), translation (p-value = 1.08 × 10(-11)), lipid biosynthetic process (p-value = 6.72 × 10(-10)), hexose metabolic process (p-value = 1.89 × 10(-04)), and actin cytoskeleton organization (p-value = 2.72 × 10(-04)). These findings will help to contribute to a better characterization of camel milk. Identified MFGM proteins from camel milk may also provide new insight into lipid droplet formation in the mammary epithelial cell.

  3. Synthesis and Biological Evaluation of Novel Aryl-2H-pyrazole Derivatives as Potent Non-purine Xanthine Oxidase Inhibitors.

    Science.gov (United States)

    Sun, Zhi-Gang; Zhou, Xiao-Jing; Zhu, Ming-Li; Ding, Wen-Ze; Li, Zhen; Zhu, Hai-Liang

    2015-01-01

    A series of aryl-2H-pyrazole derivatives were synthesized and evaluated for inhibitory activity against xanthine oxidase in vitro as potent xanthine oxidase inhibitors. Among them, 2 aryl-2H-pyrazole derivatives showed significant inhibitory activities against xanthine oxidase. Compound 19 emerged as the most potent xanthine oxidase inhibitor (IC50=9.8 µM) in comparison with allopurinol (IC50=9.5 µM). The docking study revealed that compound 19 might have strong interactions with the active site of xanthine oxidase. This compound is thus a new candidate for further development for the treatment of gout.

  4. In silico docking studies and in vitro xanthine oxidase inhibitory activity of commercially available terpenoids

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    MUTHUSWAMY UMAMAHESWARI

    2012-11-01

    Full Text Available Objective Xanthine oxidase is a highly versatile enzyme that is widely distributed among different species. The hydroxylation of purines is catalysed by xanthine oxidase and especially the conversion of xanthine to uric acid. Xanthine oxidase inhibitors are much useful, since they possess lesser side effects compared to uricosuric and anti-inflammatory agents. The present study deals with in silico and in vitro xanthine oxidase inhibitory analysis of commercially available terpenoids (bisabolol, β-caryophyllene, limonene, and α- terpinene. Methods Molecular docking studies were performed using AutoDock 4.2 and in vitro xanthine oxidase inhibitory activity was carried out using xanthine as the substrate. In addition, enzyme kinetics was performed using Lineweaver Burkplot analysis. Allopurinol, a known xanthine oxidase inhibitor was used as the standard. Results The results revealed that bisabolol exhibited a lowest binding energy value of about -7.33 kcal/mol. All other compounds showed binding energy values ranging between -7.33 to -5.87 kcal/mol which was less than the standard (-4.78 kcal/mol. In the xanthine oxidase assay, IC50 value of bisabolol was found to be 34.70 µg/ml, whereas that of allopurinol was 8.48 µg/ml. All the remaining compounds exhibited IC50 values ranging between 34.70 to 68.45 µg/ml.  In the enzyme kinetic studies, bisabolol, β-caryophyllene showed non competitive and Limonene, α- terpinene and allopurinol showed competitive type of enzyme inhibition. Conclusion It can be concluded that terpenoids could be a promising remedy for the treatment of gout and related inflammatory disorders. Further in vivo studies are required to develop potential compounds with lesser side effects.

  5. Avaliação da sensibilidade da cultura de leite do tanque para isolamento de agentes contagiosos da mastite bovina Evaluation of the sensitivity of bulk tank milk cultures for the isolation of contagious bovine mastitis pathogens

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    Maria Aparecida V. P. Brito

    1998-01-01

    ,7% para os quartos mamários. S. aureus foi isolado de todas três amostras do tanque dos rebanhos A, B e D. Somente a terceira amostra do rebanho C foi positiva para S. aureus. S agalactiae foi recuperado de todas as amostras do rebanho D, duas do rebanho C e de uma do rebanho A. Todas as amostras do tanque dos rebanhos A, B, C e D apresentaram contaminação com coliformes e somente uma das amostras coletadas na plataforma de recepção da indústria foi negativa para coliformes. Leveduras foram isoladas de 16 amostras coletadas na indústria e de todas amostras do tanque dos rebanhos A, B, C e D. Não foram isolados coliformes ou leveduras dos quartos mamários dos animais destes rebanhos, sugerindo que ocorreu contaminação do leite durante ou após a ordenha, provavelmente devido a deficiências nos processos de limpeza e higienização. A análise dos resultados das culturas do leite do tanque mostrou que o exame foi específico para detectar os patógenos contagiosos da mastite. A sensibilidade do teste aumentou quando se examinaram mais de duas amostras consecutivas.Samples of bulk tank milk from 33 herds were collected at the dairy processing plant and cultured, as a means of detecting specific (contagious bovine mastitis pathogens. Somatic cell counts (SCC were made on a Fossomatic 90. Two and three weekly consecutive samples were obtained from 13 and 12 herds, respectively. Only one sample was examined from eight herds. Three daily consecutive samples of bulk milk and individual quarter samples from all lactating cows from four herds (A, B, C and D were also examined. Milk from individual quarters were cultured on blood agar, while tank milk samples were cultured on TKT, Mannitol Salt, MacConkey agars and Sabouraud containing chloramphenicol. Staphylococcus aureus was recovered from 26 of the 33 herds sampled in the dairy processing plant. Nine of these samples also contained Streptococcus agalactiae. Nine herds had SCC above 500,000 ml-1. The remaining 23

  6. Xanthine oxidase inhibitory activity of Hungarian wild-growing mushrooms.

    Science.gov (United States)

    Ványolós, Attila; Orbán-Gyapai, Orsolya; Hohmann, Judit

    2014-08-01

    Mushrooms represent a remarkable and yet largely unexplored source of new, biologically active natural products. In this work, we report on the xanthine oxidase (XO) inhibitory activity of 47 wild-growing mushrooms native to Hungary. Aqueous and organic (n-hexane, chloroform, and 50% methanol) extracts of selected mushrooms from different families were screened for their XO inhibitory activities. Among the 188 extracts investigated, the chloroform and 50% methanol fractions proved to be the most effective. Some species exhibited high inhibitory activity, e.g., Hypholoma fasciculare (IC50  =67.76 ± 11.05 µg/mL), Suillus grevillei (IC50  =13.28 ± 1.58 µg/mL), and Tricholoma populinum (IC50  =85.08 ± 15.02 µg/mL); others demonstrated moderate or weak activity. Additional studies are warranted to characterize the compounds responsible for the XO inhibitory activity of mushroom extracts.

  7. Electrochemistry of xanthine oxidase and its interaction with nitric oxide.

    Science.gov (United States)

    Zhou, Hui; Xu, Yi; Chen, Ting; Suzuki, Iwao; Li, Genxi

    2006-02-01

    With the help of nanocrystalline TiO2, the direct electrochemistry of xanthine oxidase (XOD) was achieved and two pairs of redox waves were observed. The interaction between XOD and nitric oxide (NO) was also investigated. The experimental results reveal that NO can be reduced at a XOD-nano TiO2 film modified electrode. When the NO concentration was low, the reduced product, HNO, would inactivate the protein. However, when the NO concentration was high, HNO would continue to react with NO to form N2O2- and N3O3-, which would not inhibit XOD, and thus the amount of active protein did not decrease any further.

  8. The Influence of Xanthine Derivatives on the Oxidative Balance in terms of Emotional Stress

    Directory of Open Access Journals (Sweden)

    Sergey A. Sherstyuk

    2016-12-01

    Full Text Available This work is dedicated to study the protective action among the xanthine derivatives in the model of acute emotional stress environment. It was experimentally proved that caffeine, euphylline and pentoxyphylline have the stress protective activity in the dose of 100 mg/kg which was given twice by intragastric way. It was shown that the most active drug is pentoxyphylline, which supports peroxydant and antioxydant balance at the expense of active product of thiobarbituric acid and superoxide dismutase. The findings attest the presence of the stress protective properties in the xanthine derivatives and also suggest to use the xanthine derivatives in the diseases prevention of stress genesis.

  9. Effects of Prolactin and Leptin on Gene Expressions of Milk Proteins and Key Factors Related to Milk Protein Synthesis of Bovine Mammary Epithelial Cells%催乳素和瘦素对奶牛乳腺上皮细胞乳蛋白及乳蛋白合成信号通路关键因子基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    杜瑞平; 王春艳; 张兴夫; 高民

    2015-01-01

    protein synthesis of bovine mammary epithelial cells under pri-mary culture condition. The primary bovine mammary epithelial cells ( pBMECs) were cultured by collagenase digestion. The epithelial origin of pBMECs was identified by morphological observation, growth curve assay and specific milk protein gene expressions detection. There were four treatments with four concentrations of prolactin (0, 0.1, 1.0 and 10.0μg/mL, respectively), leptin concentration was the same (100 ng/mL), and each treatment had six replicates. Thiazole blue ( MTT) assay was used to detected the effects of prolactin and leptin on cells proliferation, and real time PCR was used to assay the effects of prolactin and leptin on gene ex-pressions of main milk proteins [α-casein,β-casein,κ-casein andβ-lactoglobulin (β-LGB) ] , Janus kinase 2 ( JAK2) , signal transduction and transcription activator 5 ( STAT5 ) and mammalian target of rapamycin ( mTOR) . The results showed that based on certain concentration of leptin ( 100 ng/mL ) , compared with 0 μg/mL prolactin treatment: the treatments of 0.1 and 1.0 μg/mL prolactin significantly promoted pBMECs proliferation (P<0.05); the treatments of 0.1 and 10.0 μg/mL prolactin significantly decreased the gene ex-pressions of αs1-casein, αs2-casein, β-casein and β-LGB(P<0.05); the treatment of 1.0 μg/mL prolactin significantly increased the gene expressions ofαs1-casein,κ-casein,β-LGB ( P<0.05);all supplemental treat-ments of prolactin exerted promotion effects on JAK2 gene expression ( P<0. 05 ); only the treatment of 1.0 μg/mL prolactin significantly increased STAT5 and mTOR gene expressions ( P<0. 05 ) . In conclusion, based on 100 ng/mL leptin in culture medium, positive effects of prolactin on gene expressions of milk pro-teins and key factors related to milk protein synthesis are observed, but the concentration should be limited in a certain range (0.1 to 1.0 μg/mL), and reverse inhibition effects emerge with high or low concentration

  10. Prevalencia y distribución espacial de brucelosis, leucosis bovina, diarrea viral bovina y rinotraqueítis infecciosa bovina a partir del análisis ELISA de estanques prediales en lecherías de la IX Región, Chile Prevalence and space distribution of brucellosis, bovine leukaemia, bovine viral diarrhea and infectious bovine rhinotracheitis by using bulk milk ELISA test in dairy herds of the IX Region, Chile

    Directory of Open Access Journals (Sweden)

    R Felmer

    2009-01-01

    of the OIE List, including foot and mouth disease and classical swine fever. However, several infectious diseases are known to remain among herds, which produce a major effect in production due to losses by abortion, decrease of fertility and what it is more important, some of them represent barriers for export and constitute a risk of zoonosis for the population. In this work, a monitoring system based on the analysis of bulk milk antibodies by means of ELISA test, was implemented to study the epidemiology and distribution of 4 of the main bovine diseases that currently affect the IX Region of Chile (brucellosis, bovine leukaemia, IBR and BVD. The system allowed the surveillance of 279 dairies, which represented 43% of the dairies registered in IX the Region, and included 19,635 milking cows (14%. With this system, a high prevalence for leukaemia (59%, IBR (76% and BVD (96% could be established, whereas it was confirmed that brucellosis is restricted to a few dairies (5%. The surveillance system coupled to the satellite geographic information analysis, allowed to establish the space distribution of these diseases in the different communes of the Region, demonstrating to be an excellent and low cost support tool for the monitoring of the diseases in the herd, which guarantees the possibility of establishing this platform in the Region and its feasibility to project it at national level.

  11. Human milk banking-facts and issues to resolve.

    Science.gov (United States)

    Corpeleijn, Willemijn E; Vermeulen, Marijn J; van Vliet, Ineke; Kruger, Caroline; van Goudoever, Johannes B

    2010-07-01

    The number of human milk banks is increasing worldwide. Although the beneficial effects of feeding premature infants with their mother's milk are well documented, less is known about the effects of feeding these infants with pasteurized donor milk. We propose a randomized trial comparing the effects of a 100% human milk-based diet (human milk supplemented with a human milk-derived fortifier) and a diet (partially) based on bovine milk. In theory, human milk has a beneficial effect on various aspects of human physiology, most of which become apparent after infancy. We therefore propose an extensive follow-up program that takes this aspect into consideration. Other issues concerning the practice of human milk banks need to be addressed as well as optimization of the feeding strategies for preterm infants.

  12. Human Milk Banking–Facts and Issues to Resolve

    Directory of Open Access Journals (Sweden)

    Willemijn E. Corpeleijn

    2010-07-01

    Full Text Available The number of human milk banks is increasing worldwide. Although the beneficial effects of feeding premature infants with their mother’s milk are well documented, less is known about the effects of feeding these infants with pasteurized donor milk. We propose a randomized trial comparing the effects of a 100% human milk-based diet (human milk supplemented with a human milk-derived fortifier and a diet (partially based on bovine milk. In theory, human milk has a beneficial effect on various aspects of human physiology, most of which become apparent after infancy. We therefore propose an extensive follow-up program that takes this aspect into consideration. Other issues concerning the practice of human milk banks need to be addressed as well as optimization of the feeding strategies for preterm infants.

  13. Diagnóstico de brucelosis bovina en leche por el Ring Test y ELISA en lecherías de la provincia de Ñuble (VIII Región Bovine brucellosis diagnosis in milk by Ring Test and ELISA in dairy farms of Ñuble (VIII Region, Chile

    Directory of Open Access Journals (Sweden)

    J. LOPEZ

    1998-01-01

    Full Text Available Se utilizaron los métodos de ELISA y Ring Test en leche para diagnosticar brucelosis bovina en estanques de recolección en frío a 60 predios lecheros de la provincia de Ñuble, VIII Región de Chile, durante los meses de noviembre y diciembre de 1995. Estos fueron estratificados en relación al número de vacas en producción de leche. Los resultados obtenidos por ambos métodos fueron similares,18 muestras positivas y 42 negativas. La diferencia se estableció en 5 muestras (8.3% que fueron clasificadas como positivas débiles al Ring Test y confirmadas como positivas por ELISA. Se determinó que la prevalencia predial de brucelosis bovina fue de un 30% en predios lecheros con estanque enfriador en la provincia de Ñuble. El métodode ELISA resultó ser una prueba confiable para diagnosticar brucelosis bovina en leches colectivas.ELISA and Ring Test were compared as diagnostic tests for bovine brucellosis in bulk tank refrigerated milk from 60 dairies in the Ñuble Province, VIII Region, Chile, from November to December. Samples were stratified according to the number of cows in the farm. Diagnosis obtained from both methods gave similar results: 18 positive and 42 negative. The difference was established in 5 samples (8.3% that were classified as weak positive to the Ring Test and confirmed as positives by ELISA. Furthermore, it was determined that there was 30% prevalence of bovine brucellosis in dairies with refrigerated tanks in the Ñuble province. The ELISA proved to be a reliable diagnostic test of bovine brucellosis in bulk milk

  14. Milk Allergy

    Science.gov (United States)

    ... Events Blog Media Shop Alerts Donate About Food Allergies Home About Food Allergy Food Allergy Basics Facts ... Registration Create Your Own Events Educational Events Milk Allergy Allergy to cow’s milk is the most common ...

  15. The use of successive milking as a procedure for the elimination of infection on due to Prototheca spp in cases of bovine clinical mastitis. (A case report / Utilização de ordenhas sucessivas como procedimento para eliminação de infecção por prototheca spp de casos de mastite clínica bovina (relato de caso

    Directory of Open Access Journals (Sweden)

    Márcia Kiyoe Shimada

    2000-01-01

    Full Text Available This work relates the case of two animals (Black and White Holstein from a bovine milking herd, from north of Paraná State, in which mastitis due Prototheca spp was diagnosed. As a procedure for the infection elimination, it was opted for the use of successive milking performed six times daily, during eight days. The animals were followed up through mycological cultures from milk Samples after 15, 30 and 60 days from the beginning of the procedure. The clinical signs regressed after three days and the mycologicai cultures were negative after the 15th day.O presente trabalho relata um caso de dois animais HPB de um rebanho bovino leiteiro, da região norte do Estado do Paraná, dos quais diagnosticou-se mastite clínica por Prototheca spp. Como procedimento para eliminação da infecção optou-se peta utilização de ordenhas sucessivas praticadas seis vezes diariamente durante oito dias. Os animais foram acompanhados através de culturas micológicas das amostras de leite após 15, 30 e 60 dias do inicio do procedimento. Os sinais clínicos regrediram após três dias, e a partir do 15° dia as culturas foram negativas.

  16. [Comparison of the function and conformation of human β-casein and bovine β-casein by spectroscopic study].

    Science.gov (United States)

    Liu, Wei; Li, Meng; Ren, Hao-wei; Liu, Ning

    2014-12-01

    β-Casein was the main component of human milk casein, but the content of β-casein in the bovine milk was less. The difference in β-casein content of the two samples was one of the reasons why human milk is more digestible than bovine milk. Studying the differences of structure and function in human and bovine milk β-casein can help us develop a new human milk simulated infant formula which will be more suitable for the infant gut. The UV spectrophotometer was used to study the solubility, sulfhydryl and emulsification of human milk β-casein and bovine milk β-casein, Fluorescence spectroscopy and the infrared spectroscopy were used to study the structural characteristics of human milk β-casein and bovine milk β-casein. The two samples shared a similar isoelectric point (pH 4.0~5.0), the solubility of human milk β-casein (10.83%) was lower than which in bovine milk β-casein (11.83%) near the pI, while it was higher when it deviated the pI. The emulsion ability (110~140 m2 · g(-1)) of human milk β-casein was higher than that in bovine milk β-casein (70~130 m2 · g(-1)) and surface sulfhydryl group (SH) of two kinds of milk protein were similar [(18.47±0.08) μmol · g(-1) and (18.67±0.17) μmol · g(-1)]. The total sulfhydryl group [(47.46±0.23) μmol · g(-1)] in bovine milk β-casein was more than that in human milk β-casein [(26.17±0.12) μmol · g(-1)]. Functional groups in two samples were similar and they both contained beta sheet, human milk β-casein had less H-bond and internal hydrophobic than bovine milk β-casein. The results showed that the two samples had similar functional groups, while human milk β-casein had much less secondary structure such as α-helix and β-sheet, a looser tertiary structure and a better interfacial activity.

  17. Efeito da utilização da somatotropina bovina recombinante (bST sobre a produção de leite em búfalas Effect of recombinant bovine somatotropin (bST utilization on milk production from buffaloes

    Directory of Open Access Journals (Sweden)

    André Mendes Jorge

    2002-06-01

    Full Text Available O objetivo deste trabalho foi estudar o resultado da utilização da somatotropina bovina recombinante (bST atuando na produção de leite em búfalas da raça Murrah. Empregaram-se 28 búfalas multíparas da raça Murrah, divididas em dois grupos homogêneos de 14 animais, recebendo os seguintes tratamentos: Grupo 1 ¾ Controle (solução salina e Grupo 2 ¾ 500 mg de bST/cabeça a cada 14 dias, durante 7 meses. Búfalas tratadas com bST exibiram incrementos de 48,52%, 32,80% e 32,80% nas produções total de leite, corrigida, depois, para 4% de gordura e média diária, respectivamente. A somatotropina elevou a produção total de gordura sem alterar a porcentagem dela no leite. A administração de bST não afetou a porcentagem de proteína do leite todavia, a produção total de proteína foi aumentada. Quanto à duração da lactação, o tratamento com bST diferiu do controle, o que demonstra a maior persistência da lactação de búfalas tratadas com bST.The objective of this work was to study the effect of recombinant bovine somatotropin (bST utilization on milk production from Murrah buffaloes. Twenty eight multiparous Murrah buffaloes were used and divided into two homogeneous groups of 14 animals, receiving the following treatments: Group 1 ¾ Control (salt solution and Group 2 ¾ 500 mg of bST/head every 14 days during 7 months. Buffaloes treated with bST presented increase of 48.52%, 32.80% and 32.80% on total milk yield, adjusted to 4% of fat and average daily milk yield, respectively. Somatotropin increased total fat milk yield without alter fat percentage of milk. Administration of bST did not affect protein percentage of milk while total protein milk yield increased. As for the lactating period, the treatment with bST differed of the control, what might have denoted in larger persistence of the lactation from buffaloes treated with bST.

  18. Structure, function and nutritional potential of milk osteopontin

    DEFF Research Database (Denmark)

    Christensen, Brian Søndergaard; Sørensen, Esben Skipper

    2016-01-01

    of milk OPN. At the same time, plasma OPN levels have been shown to be elevated in some types of cancer, and OPN has been suggested as a potential diagnostic marker for cancer. OPN exists in several different isoforms in vivo, of which presumably only a minority is directly or indirectly implicated...... in cancer related events. In this article, we review the differences between milk-derived OPN and OPN derived from transformed cells and compare the structure of OPN from human and bovine milk. Furthermore, current knowledge about the function of OPN in milk and recent findings about the effect of orally......Osteopontin (OPN) is a multifunctional protein present in most tissues and body fluids, with the highest concentrations found in milk. Processes for isolation of OPN from bovine milk for use in infant formula have been developed and studies have investigated the effects of oral administration...

  19. Effect of carryover and presampling procedures on the results of real-time PCR used for diagnosis of bovine intramammary infections with Streptococcus agalactiae at routine milk recordings

    DEFF Research Database (Denmark)

    Mahmmod, Yasser; Mweu, Marshal Mutinda; Nielsen, Søren Saxmose;

    2014-01-01

    (Ct) cut-offs by estimating the between-cow correlation while accounting for the milking order, and (2) evaluate the effect of aseptic presampling procedures (PSP) on PCR-positivity at the different Ct-value cut-offs. The study was conducted in four herds with conventional milking parlours at routine...

  20. [Milk fever].

    Science.gov (United States)

    Dumont, M

    1989-05-01

    Infectious complications following delivery were, in the past, attributed to "milk fever": these were milk congestion, milk deposits, rancid milk, etc., that were held responsible. The milk was reabsorbed into the blood of the patient and settled in the peritoneum ("milk peritonitis"), in the broad ligaments (pelvic abscess), in the thighs (phlebitis) and also in the breasts (breast abscess). This belief, originated by Aristotle, was accepted by excellent authors like Andre Levret (1703-1780), one of the most famous French obstetricians and Nicolas Puzos, at the same time. More recently, authors alluded to it and blamed "milk fever" for being at the origin of dramatic pictures which they described in their novels, like Victor Hugo and Guy de Maupassant, for instance.

  1. Amiata donkey milk chain: animal health evaluation and milk quality

    Directory of Open Access Journals (Sweden)

    Giuseppe Ragona

    2016-08-01

    Full Text Available This study presents an investigation of Amiata donkey health and quality of milk for human consumption. Thirty-one lactating dairy jennies were examined. The following samples were collected: faecal samples from the rectum of animals for parasitological examination; cervical swabs for the detection of bacteria causing reproductive disorders; and blood samples for serological diagnosis of main zoonotic (Brucella spp., Leptospira spp. and donkey abortion agents (Brucella spp., Leptospira spp., Salmonella abortus equi, Equine viral arterithis virus, Equine herpesvirus type 1. In addition, individual milk samples were collected and analysed for mastitis- causing pathogens and milk quality. Regarding animal health, we detected a high prevalence of strongyle parasites in donkeys. It is very important to tackle parasitic diseases correctly. Selective control programmes are preferable in order to reduce anthelmintic drug use. For dairy donkeys, withdrawal periods from anthelmintic drugs need to be carefully managed, in accordance with EU and national regulations. The isolation of Staphylococcus aureus in milk highlights the importance of preventing contamination during milking, by adopting appropriate hygiene and safety practices at a farm level. Lysozyme activity was high compared to cow’s milk, contributing to the inhibitory activity against certain bacteria. Donkey milk was characterised by high lactose content, low caseins, low fat, higher levels of unsaturated fatty acids compared to ruminant milks. Unsaturated fatty acids and omega 3 fatty acids in particular have become known for their beneficial health effect, which is favourable for human diet. These characteristics make it suitable for infants and children affected by food intolerance/ allergies to bovine milk proteins and multiple food allergies as well as for adults with dyslipidemias. It is also recommended to prevent cardiovascular diseases.

  2. Amiata Donkey Milk Chain: Animal Health Evaluation and Milk Quality

    Science.gov (United States)

    Ragona, Giuseppe; Corrias, Franco; Benedetti, Martina; Paladini, Maria; Salari, Federica; Altomonte, lolanda; Martini, Mina

    2016-01-01

    This study presents an investigation of Amiata donkey health and quality of milk for human consumption. Thirty-one lactating dairy jennies were examined. The following samples were collected: faecal samples from the rectum of animals for parasitological examination; cervical swabs for the detection of bacteria causing reproductive disorders; and blood samples for serological diagnosis of main zoonotic (Brucella spp., Leptospira spp.) and donkey abortion agents (Brucella spp., Leptospira spp., Salmonella abortus equi, Equine viral arterithis virus, Equine herpesvirus type 1). In addition, individual milk samples were collected and analysed for mastitis-causing pathogens and milk quality. Regarding animal health, we detected a high prevalence of strongyle parasites in donkeys. It is very important to tackle parasitic diseases correctly. Selective control programmes are preferable in order to reduce anthelmintic drug use. For dairy donkeys, withdrawal periods from anthelmintic drugs need to be carefully managed, in accordance with EU and national regulations. The isolation of Staphylococcus aureus in milk highlights the importance of preventing contamination during milking, by adopting appropriate hygiene and safety practices at a farm level. Lysozyme activity was high compared to cow’s milk, contributing to the inhibitory activity against certain bacteria. Donkey milk was characterised by high lactose content, low caseins, low fat, higher levels of unsaturated fatty acids compared to ruminant milks. Unsaturated fatty acids and omega 3 fatty acids in particular have become known for their beneficial health effect, which is favourable for human diet. These characteristics make it suitable for infants and children affected by food intolerance/allergies to bovine milk proteins and multiple food allergies as well as for adults with dyslipidemias. It is also recommended to prevent cardiovascular diseases. PMID:27853717

  3. Amiata Donkey Milk Chain: Animal Health Evaluation and Milk Quality.

    Science.gov (United States)

    Ragona, Giuseppe; Corrias, Franco; Benedetti, Martina; Paladini, Maria; Salari, Federica; Altomonte, Lolanda; Martini, Mina

    2016-06-03

    This study presents an investigation of Amiata donkey health and quality of milk for human consumption. Thirty-one lactating dairy jennies were examined. The following samples were collected: faecal samples from the rectum of animals for parasitological examination; cervical swabs for the detection of bacteria causing reproductive disorders; and blood samples for serological diagnosis of main zoonotic (Brucella spp., Leptospira spp.) and donkey abortion agents (Brucella spp., Leptospira spp., Salmonella abortus equi, Equine viral arterithis virus, Equine herpesvirus type 1). In addition, individual milk samples were collected and analysed for mastitis-causing pathogens and milk quality. Regarding animal health, we detected a high prevalence of strongyle parasites in donkeys. It is very important to tackle parasitic diseases correctly. Selective control programmes are preferable in order to reduce anthelmintic drug use. For dairy donkeys, withdrawal periods from anthelmintic drugs need to be carefully managed, in accordance with EU and national regulations. The isolation of Staphylococcus aureus in milk highlights the importance of preventing contamination during milking, by adopting appropriate hygiene and safety practices at a farm level. Lysozyme activity was high compared to cow's milk, contributing to the inhibitory activity against certain bacteria. Donkey milk was characterised by high lactose content, low caseins, low fat, higher levels of unsaturated fatty acids compared to ruminant milks. Unsaturated fatty acids and omega 3 fatty acids in particular have become known for their beneficial health effect, which is favourable for human diet. These characteristics make it suitable for infants and children affected by food intolerance/allergies to bovine milk proteins and multiple food allergies as well as for adults with dyslipidemias. It is also recommended to prevent cardiovascular diseases.

  4. Xanthine oxidase activity regulates human embryonic brain cells growth

    Directory of Open Access Journals (Sweden)

    Kevorkian G. A.

    2011-10-01

    Full Text Available Aim. Involvement of Xanthine Oxidase (XO; EC1.1.3.22 in cellular proliferation and differentiation has been suggested by the numerous investigations. We have proposed that XO might have undoubtedly important role during the development, maturation as well as the death of human embryos brain cells. Methods. Human abortion material was utilized for the cultivation of brain cells (E90. XO activity was measured by the formation of uric acid in tissue. Cell death was detected by the utility of Trypan Blue dye. Results. Allopurinol suppressed the XO activity in the brain tissue (0.12 ± 0.02; 0.20 ± 0.03 resp., p < 0.05. On day 12th the number of cells in the culture treated with the Allopurinol at the early stage of development was higher in comparison with the Control (2350.1 ± 199.0 vs 2123 ± 96 and higher in comparison with the late period of treatment (1479.6 ± 103.8, p < < 0.05. In all groups, the number of the dead cells was less than in Control, indicating the protective nature of Allopurinol as an inhibitor of XO. Conclusions. Allopurinol initiates cells proliferation in case of the early treatment of the human brain derived cell culture whereas at the late stages it has an opposite effect.

  5. Xanthine oxidase inhibitory activity of compounds from Chythrantus claneianus

    Directory of Open Access Journals (Sweden)

    Anar Sahib Gojayev

    2013-03-01

    Full Text Available Phytochemical investigation of the stem bark and the trunk of Chythrantus claneianus led to the isolation of six known compounds named β-sitosterol (1, umbelliferone (2, scopoletin (3, benjaminamide (4, β-sitosterol-3-O-β-D-glucopyranoside (5 and Panconoside B (6. All these compounds were isolated for the first time from this plant species. The chemical structures of isolates were elucidated on the basis of 1 and 2 D-NMR spectra and other spectroscopic techniques including UV–vis, FT-IR, HR-ESIMS and HR-FABMS. The isolates were tested in vitro for their inhibitory properties towards xanthine oxidase enzyme. Compounds 2, 3 and 6 showed weak inhibi-tory activities on the enzyme with IC50 values ranging from 307 µM for com-pound 6 to 475 µM for compound 3, while the extract and compounds 1, 4 and 5 showed extremely weak activities with inhibition percentage less than 50%.

  6. Cow's milk and goat's milk.

    Science.gov (United States)

    Turck, Dominique

    2013-01-01

    Cow's milk is increasingly suggested to play a role in the development of chronic degenerative, non-communicable disorders whereas goat's milk is advocated as having several health benefits. Cow's milk is a rich and cheap source of protein and calcium, and a valuable food for bone health. Despite their high content in saturated fats, consumption of full-fat dairy products does not seem to cause significant changes in cardiovascular disease risk variables. Early introduction of cow's milk is a strong negative determinant of iron status. Unmodified cow's milk does not meet nutritional requirements of infants although it is acceptable to add small volumes of cow's milk to complementary foods. Cow's milk protein allergy has a prevalence ranging from 2 to 7%, and the age of recovery is usually around 2-3 years. The evidence linking cow's milk intake to a later risk of type 1 diabetes or chronic degenerative, non-communicable disorders (obesity, metabolic syndrome, type 2 diabetes, hypertension) is not convincing. Milk probably protects against colorectal cancer, diets high in calcium are a probable cause of prostate cancer, and there is limited evidence suggesting that high consumption of milk and dairy products increases the risk for prostate cancer. There is no evidence to support the use of a cow's milk-free diet as a primary treatment for individuals with autistic spectrum disorders. Unmodified goat's milk is not suitable for infants because of the high protein and minerals content and of a low folate content. Goat's milk has no clear nutritional advantage over cow's milk and is not less allergenic. The European Food Safety Authority recently stated that proteins from goat's milk can be suitable as a protein source for infant and follow-on formula, provided the final product complies with the compositional criteria laid down in Directive 2006/141/EC.

  7. Kajian Fisiologis Penggunaan Bovine Somatotropin (bST Pada Sapi Pra Afkir

    Directory of Open Access Journals (Sweden)

    Dzarnisa Araby

    2009-10-01

    Full Text Available Studies of physiological bovine somatotropin (bST on post lactating dairy ABSTRACT. To increase of milk production nationally with used in dairy cattle business can assist to increase milk production, eighteen post lactating dairy cows in the highland Cipelang Bogor, were used to study the effect used of bovine somatotropin and lactating time. The experimental cows were assigned into a Randomized Block Design with a 3x2 factorial arrangement. The first factor was using of somatotropin (bSTwith three levels (non bST injection, biweekly injection and three weekly injection. The second factor was lactating time with two levels (4th lactating time and 6 th lactating time Parameters measured were heart rate, respiration frequency, rectal temperature, milk production efficiency, milk production, 4% FCM (fat corrected milk., Milk composition, weigh gain, milk quality consist of protein, fat, pH. Bovine somatotropin significantly increased heart rate and respiration rate. Also bovine somatotropin injection at 4th lactating time significantly increased milk production. There were an interaction between bST dan lactating time on milk production and weight gain. Bovine somatotropin injection biweekly in cows on 4th lactating time ration increased milk production by 16-26 %, but injection in cows 6th lactating time increased milk production by 8-18 % combination with somatotropin doze 250/ml/14 days. Somatothropine supplementation was injection biweekly and three weekly did significantly affect to milk production, body temperature, heart rate, and respiration rate however in normal physiology . Bovine somatotropine can increase post lactating dairy production interval 14 days better than 21 days.

  8. Preparation of a Xanthine Sensor Based on the Immobilization of Xanthine Oxidase on a Chitosan Modified Electrode by Cross-linking

    Institute of Scientific and Technical Information of China (English)

    刘玉革; 李伟明; 魏长宾; 吕玲玲

    2012-01-01

    Here in this paper, xanthine oxidase (XOD) was immobilized onto the chitosan (CHT) modified electrode by a simple way of cross-linking with glutaraldehyde (GTD) and 3-aminopropyltriethoxysilane (KH). The electrode displayed a sharp peak to the oxidation of xanthine at a potential about 0.67 V and the optimum of pH for determination was investigated. Under the optimum conditions, the biosensor fabricated on the KH/GTD/XOD/CHT modified electrode showed excellent response to the oxidation of xanthine within the range of 0.5 to 18 μmol/L with a low detection limit of 0.0215 μmol/L, a good stability and a high selectivity. The sensor can also be used for the determination of hypoxantbine. The electrochemical results indicated that the immobilized enzyme still retained its biological activity and this provided a new way for the construction of biosensors and determination of xanthine.

  9. Evaluation of mineral content and heavy metals of dromedary camel milk in Iran

    Directory of Open Access Journals (Sweden)

    Mahdieh MOSTAFIDI

    Full Text Available Abstract The aim of this study was to determine the amount of major mineral compounds and heavy metals of camel milk in Iran. For this purpose camel milk samples were collected from seven regions of Iran include Qazvin, Golestan, Semnan, Sistan-Baluchestan, Khuzestan, Bushehr and Tehran. The samples were analyzed using Inductively Coupled Plasma-Optical Emission Spectrometer (ICP-OES method. The results showed that among the mineral contents, iron and zinc of camel milk were greater than bovine milk. Based on the codex standard 193-2007 standards, the maximum acceptable limit for lead and cadmium is 20 µg/kg and 10 µg/kg, respectively. The results of this study showed that the measured amounts of lead, cadmium and nickel in all samples were less than the acceptable limit for bovine milk. Bovine milk and dairy products are a poor source of iron, while the obtained data revealed that camel milk is a major source of minerals, especially iron. The camel milk’s iron was 10 times more than bovine milk. However, variations in mineral content in camel milk could be due to feed, stage of lactation, milk collection time, drought conditions, environmental conditions and associated analytical methods. Camel milk recommended as a valuable source of food for the human.

  10. Purification of RNA from milk whey.

    Science.gov (United States)

    Izumi, Hirohisa; Kosaka, Nobuyoshi; Shimizu, Takashi; Sekine, Kazunori; Ochiya, Takahiro; Takase, Mitsunori

    2013-01-01

    MicroRNAs (miRNAs) are small regulatory RNA molecules that modulate specific target mRNAs and play very important roles in physiological processes. They were recently detected in body fluids such as blood, urine, saliva, and milk. These body fluid miRNAs have been studied thoroughly as potential diagnostic biomarkers. However, there have been few studies of milk miRNAs, and their roles are not clearly understood. Milk is the only nutritional source for newborn infants, and bovine milk is used widely as a dairy product. Thus, it is important to study milk miRNAs. In general, body fluid RNA concentrations are extremely low and of diverse existence types. In this chapter, we compare two silica membrane column-based RNA purification kits, and also compare RNA obtained directly from whey with that isolated from whey-derived exosomes.

  11. Selective tracheal relaxation and phosphodiesterase-IV inhibition by xanthine derivatives.

    Science.gov (United States)

    Miyamoto, K; Kurita, M; Ohmae, S; Sakai, R; Sanae, F; Takagi, K

    1994-05-17

    The effects of substitutions in the xanthine nucleus on tracheal relaxant activity, atrium chronotropic activity, adenosine A1 affinity, and inhibitory activities on cyclic AMP-phosphodiesterase isoenzymes in guinea pigs were studied. Substitution with a long alkyl chain at the N1-position of xanthine nucleus increased the tracheal relaxant activity without leading to positive chronotropic action, and long alkyl chains at the N3-position increased both activities. N7-substitutions with n-propyl and 2'-oxopropyl groups, such as in denbufylline, increased bronchoselectivity. N7-substitution decreased the adenosine A1 affinity, but substitution at either the N1- or N3-position increased it. The bronchorelaxant activity of xanthine derivatives was closely correlated with their inhibition of phosphodiesterase-IV, but not with their adenosine A1 affinity; the positive chronotropic effects were related to their inhibition of phosphodiesterase-III. This study confirms that the bronchorelaxation of xanthine derivatives is mediated by inhibition of the isoenzyme phosphodiesterase-IV. The results of structure-activity analysis suggest that substitutions at the N1- and N7-positions should be tried in the development of xanthine derivatives that are selective bronchodilators and phosphodiesterase-IV inhibitors.

  12. A retrospective analysis of the infectious bovine rhinotracheitis (bovine herpes virus-1) surveillance program in Norway using Monte Carlo simulation models

    DEFF Research Database (Denmark)

    Paisley, Larry; Tharaldsen, J.; Jarp, J.

    2001-01-01

    Serological surveillance for antibodies against bovine herpes virus type I (BHV-1) which causes infectious bovine rhinotracheitis and infectious pustular vulvovaginitis has been carried out since 1992 in Norway. Since 1993 (when a single infected herd was detected) all bulk-milk and pooled-serum...

  13. Bovine Mastitis: Frontiers in Immunogenetics

    Directory of Open Access Journals (Sweden)

    Kathleen eThompson-Crispi

    2014-10-01

    Full Text Available Mastitis is one of the most prevalent and costly diseases in the dairy industry with losses attributable to reduced milk production, discarded milk, early culling, veterinary services, and labor costs. Typically, mastitis is an inflammation of the mammary gland most often, but not limited to, bacterial infection, and is characterized by the movement of leukocytes and serum proteins from the blood to the site of infection. It contributes to compromised milk quality and the potential spread of antimicrobial resistance if antibiotic treatment is not astutely applied. Despite the implementation of management practises and genetic selection approaches, bovine mastitis control continues to be inadequate. However, some novel genetic strategies have recently been demonstrated to reduce mastitis incidence by taking advantage of a cow’s natural ability to make appropriate immune responses against invading pathogens. Specifically, dairy cattle with enhanced and balanced immune responses have a lower occurrence of disease, including mastitis, and they can be identified and selected for using the High Immune Response (HIR technology. Enhanced immune responsiveness is also associated with improved response to vaccination, increased milk and colostrum quality. Since immunity is an important fitness trait, beneficial associations with longevity and reproduction are also often noted. This review highlights the genetic regulation of the bovine immune system and its vital contributions to disease resistance. Genetic selection approaches currently used in the dairy industry to reduce the incidence of disease are reviewed, including the HIR technology, genomics to improve disease resistance or immune response, as well as the Immunity+TM sire line. Improving the overall immune responsiveness of cattle is expected to provide superior disease resistance, increasing animal welfare and food quality while maintaining favourable production levels to feed a growing

  14. Estimation of genetic parameters and detection of quantitative trait loci for metabolites in Danish Holstein milk

    DEFF Research Database (Denmark)

    Buitenhuis, Albert Johannes; Sundekilde, Ulrik; Poulsen, Nina Aagaard;

    2013-01-01

    Small components and metabolites in milk are significant for the utilization of milk, not only in dairy food production but also as disease predictors in dairy cattle. This study focused on estimation of genetic parameters and detection of quantitative trait loci for metabolites in bovine milk....... For this purpose, milk samples were collected in mid lactation from 371 Danish Holstein cows in first to third parity. A total of 31 metabolites were detected and identified in bovine milk by using 1H nuclear magnetic resonance (NMR) spectroscopy. Cows were genotyped using a bovine high-density single nucleotide...... polymorphism (SNP) chip. Based on the SNP data, a genomic relationship matrix was calculated and used as a random factor in a model together with 2 fixed factors (herd and lactation stage) to estimate the heritability and breeding value for individual metabolites in the milk. Heritability was in the range of 0...

  15. De novo analysis of receptor binding affinity data of xanthine adenosine receptor antagonists.

    Science.gov (United States)

    Dalpiaz, A; Gardenghi, A; Borea, P A

    1995-03-01

    The receptor binding affinity data to adenosine A1 and A2 receptors of a wide series of xanthine derivatives have been analyzed by means of the Free-Wilson model. The analysis of the individual group contribution shows, for both A1 and A2 receptors, the primary importance of the presence of bulky substituents at position 8 for an optimum receptor binding. Moreover, considering the different aij contributions of bulky substituents at position 8 for affinity to A1 with respect to A2 receptors, this position appears to be the most important for the synthesis of highly A1 selective xanthine derivatives. Moreover the analysis of group contributions for other substitution positions of the xanthine moiety allows to state that suitable substitutions at positions 3 and 7 could confer some degree of A2 selectivity.

  16. QSAR and SAR studies on the reduction of some aromatic nitro compounds by xanthine oxidase.

    Science.gov (United States)

    Thakur, Mamta; Thakur, Abhilash; Balasubramanian, Krishnan

    2006-01-01

    This work describes QSAR and SAR studies on the reduction of 27 aromatic nitro compounds by xanthine oxidase using both distance-based topological indices and quantum molecular descriptors along with indicator parameters. The application of a multiple linear regression analysis indicated that a combination of distance-based topological indices with the ad hoc molecular descriptors and the indicator parameters yielded a statistically significant model for the activity, log K (the reduction of aromatic nitro compounds by xanthine oxidase). The final selection of a potential aromatic nitro compound for the reduction by xanthine oxidase is made by quantum molecular modeling. We have found that, among the various parameters, the quantum Mulliken charge parameters on the fourth atom or para position relative to the nitro group correlated best with the activity.

  17. Vibrational spectral investigation on xanthine and its derivatives—theophylline, caffeine and theobromine

    Science.gov (United States)

    Gunasekaran, S.; Sankari, G.; Ponnusamy, S.

    2005-01-01

    A normal coordinate analysis has been carried out on four compounds having a similar ring structure with different side chain substitutions, which are xanthine, caffeine, theophylline, and theobromine. Xanthine is chemically known as 2,6-dihydroxy purine. Caffeine, theophylline and theobromine are methylated xanthines. Considering the methyl groups as point mass, the number of normal modes of vibrations can be distributed as Γ vib=27 A'+12 A″ based on C s point group symmetry associated with the structures. In the present work 15 A' and 12 A″ normal modes are considered. A new set of orthonormal symmetry co-ordinates have been constructed. Wilson's F- G matrix method has been adopted for the normal coordinate analysis. A satisfactory vibrational band assignment has been made by employing the FTIR and FT Raman spectra of the compounds. The potential energy distribution is calculated with the arrived values of the force constants and hence the agreement of the frequency assignment has been checked.

  18. Xanthine oxidase activity and free radical generation in patients with sepsis syndrome

    DEFF Research Database (Denmark)

    Galley, H F; Davies, Michael Jonathan; Webster, N R

    1996-01-01

    OBJECTIVE: To determine xanthine oxidase activity, free radical concentrations, and lipid peroxidation in patients with sepsis syndrome compared with noninfected critically ill patients. DESIGN: A prospective observational study. SETTING: A nine-bed intensive care unit in a university teaching......). CONCLUSIONS: Patients with sepsis have xanthine oxidase activation, high free-radical concentrations, and evidence of free radical damage. The finding that xanthine oxidase activity was lower in those patients who died, coupled with increased lactate concentrations implies more severe ischemia with incomplete...... to the Acute Physiology and Chronic Health Evaluation (APACHE) II score or to the presence of organ dysfunction. The mean ascorbyl radical concentration (arbitrary units) determined by electron paramagnetic resonance following spin trapping was increased in patients compared with healthy subjects (p

  19. Development of a New Reporter Gene System-dsRed/Xanthine Phosphoribosyltransferase-Xanthine for Molecular Imaging of Processes Behind the Intact Blood-Brain Barrier

    Directory of Open Access Journals (Sweden)

    Mikhail Doubrovin

    2003-04-01

    Full Text Available We report the development of a novel dual-modality fusion reporter gene system consisting of Escherichia coli xanthine phosphoribosyltransferase (XPRT for nuclear imaging with radiolabeled xanthine and Discosoma red fluorescent protein for optical fluorescent imaging applications. The dsRed/XPRT fusion gene was successfully created and stably transduced into RG2 glioma cells, and both reporters were shown to be functional. The level of dsRed fluorescence directly correlated with XPRT enzymatic activity as measured by ribophosphorylation of [14C]-xanthine was in vitro (Ki = 0.124 ± 0.008 vs. 0.00031 ± 0.00005 mL/min/g in parental cell line, and [*]-xanthine octanol/water partition coefficient was 0.20 at pH = 7.4 (logP = 0.69, meeting requirements for the blood-brain barrier (BBB penetrating tracer. In the in vivo experiment, the concentration of [* C]-xanthine in the normal brain varied from 0.20 to 0.16 + 0.05% dose/g under 0.87 + 0.24% dose/g plasma radiotracer concentration. The accumulation in vivo in the transfected flank tumor was to 2.4 ± 0.3% dose/g, compared to 0.78 ± 0.02% dose/g and 0.64 ± 0.05% dose/g in the control flank tumors and intact muscle, respectively. [14C]-Xanthine appeared to be capable of specific accumulation in the transfected infiltrative brain tumor (RG2-dsRed/XPRT, which corresponded to the 585 nm fluorescent signal obtained from the adjacent cryosections. The images of endogenous gene expression with the “sensory system” have to be normalized for the transfection efficiency based on the “beacon system” image data. Such an approach requires two different “reporter genes” and two different “reporter substrates.” Therefore, the novel dsRed/XPRT fusion gene can be used as a multimodality reporter system in the biological applications requiring two independent reporter genes, including the cells located behind the BBB.

  20. Determination of serum adenosine deaminase and xanthine oxidase levels in patients with crimean-congo hemorrhagic fever

    Directory of Open Access Journals (Sweden)

    V. Kenan Celik

    2010-01-01

    Full Text Available OBJECTIVE: Crimean-Congo hemorrhagic fever is an acute viral hemorrhagic fever with a high mortality rate. Despite increasing knowledge about hemorrhagic fever viruses, little is known about the pathogenesis of Crimean-Congo hemorrhagic fever. In this study, we measured serum adenosine deaminase and xanthine oxidase levels in Crimean-Congo hemorrhagic fever patients. METHODS: Serum adenosine deaminase levels were measured with a sensitive colorimetric method described by Giusti and xanthine oxidase levels by the method of Worthington in 30 consecutive hospitalized patients (mean age 42.6 ± 21.0. Laboratory tests confirmed their diagnoses of Crimean-Congo hemorrhagic fever. Thirty-five subjects (mean age 42.9 ± 19.1 served as the control group. RESULTS: There was a significant difference in adenosine deaminase and xanthine oxidase levels between cases and controls (p0.05. CONCLUSION: Adenosine deaminase and xanthine oxidase levels were increased in patients with Crimean-Congo hemorrhagic fever. Elevated serum xanthine oxidase activity in patients with Crimean-Congo hemorrhagic fever may be associated with reactive oxygen species generated by the xanthine/xanthine oxidase system during inflammatory responses. In addition, elevated lipid peroxidation may contribute to cell damage and hemorrhage. The association of cell damage and hemorrhage with xanthine oxidase activity should be further investigated in large-scale studies.

  1. The extraordinary specificity of xanthine phosphoribosyltransferase from Bacillus subtilis elucidated by reaction kinetics, ligand binding, and crystallography

    DEFF Research Database (Denmark)

    Arent, Susan; Kadziola, Anders; Larsen, Sine;

    2006-01-01

    Xanthine phosphoribosyltransferase (XPRTase) from Bacillus subtilis is a representative of the highly xanthine specific XPRTases found in Gram-positive bacteria. These XPRTases constitute a distinct subclass of 6-oxopurine PRTases, which deviate strongly from the major class of H(X)GPRTases with ...

  2. Thermostable Xanthine Oxidase Activity from Bacillus pumilus RL-2d Isolated from Manikaran Thermal Spring: Production and Characterization.

    Science.gov (United States)

    Sharma, Nirmal Kant; Thakur, Shikha; Thakur, Neerja; Savitri; Bhalla, Tek Chand

    2016-03-01

    Xanthine oxidase is an important enzyme of purine metabolism that catalyzes the hydroxylation of hypoxanthine to xanthine and then xanthine to uric acid. A thermostable xanthine oxidase is being reported from a thermophilic organism RL-2d isolated from the Manikaran (Kullu) hot spring of Himachal Pradesh (India). Based on the morphology, physiological tests, and 16S rDNA gene sequence, RL-2d was identified as Bacillus pumilus. Optimization of physiochemical parameters resulted into 4.1-fold increase in the xanthine oxidase activity from 0.051 U/mg dcw (dry cell weight) to 0.209 U/mg dcw. The xanthine oxidase of B. pumilus RL-2d has exhibited very good thermostability and its t1/2 at 70 and 80 °C were 5 and 1 h, respectively. Activity of this enzyme was strongly inhibited by Hg(2+), Ag(+) and allopurinol. The investigation showed that B. pumilus RL-2d exhibited highest xanthine oxidase activity and remarkable thermostability among the other xanthine oxidases reported so far.

  3. Hypouricaemic action of mangiferin results from metabolite norathyriol via inhibiting xanthine oxidase activity.

    Science.gov (United States)

    Niu, Yanfen; Liu, Jia; Liu, Hai-Yang; Gao, Li-Hui; Feng, Guo-Hua; Liu, Xu; Li, Ling

    2016-09-01

    Context Mangiferin has been reported to possess a potential hypouricaemic effect. However, the pharmacokinetic studies in rats showed that its oral bioavailability was only 1.2%, suggesting that mangiferin metabolites might exert the action. Objective The hypouricaemic effect and the xanthine oxidase inhibition of mangiferin and norathyriol, a mangiferin metabolite, were investigated. Inhibition of norathyriol analogues (compounds 3-9) toward xanthine oxidase was also evaluated. Materials and methods For a dose-dependent study, mangiferin (1.5-6.0 mg/kg) and norathyriol (0.92-3.7 mg/kg) were administered intragastrically to mice twice daily for five times. For a time-course study, mice received mangiferin and norathyriol both at a single dose of 7.1 μmol/kg. In vitro, inhibition of test compounds (2.4-2.4 mM) against xanthine oxidase activity was evaluated by the spectrophotometrical method. The inhibition type was identified from Lineweaver-Burk plots. Results Norathyriol (0.92, 1.85 and 3.7 mg/kg) dose dependently decreased the serum urate levels by 27.0, 33.6 and 37.4%, respectively. The action was more potent than that of mangiferin at the low dose, but was equivalent at the higher doses. Additionally, the hypouricaemic action of them exhibited a time dependence. In vitro, norathyriol markedly inhibited the xanthine oxidase activities, with the IC50 value of 44.6 μM, but mangiferin did not. The kinetic studies showed that norathyriol was an uncompetitive inhibitor by Lineweaver-Burk plots. The structure-activity relationships exhibited that three hydroxyl groups in norathyriol at the C-1, C-3 and C-6 positions were essential for maintaining xanthine oxidase inhibition. Discussion and conclusion Norathyriol was responsible for the hypouricaemic effect of mangiferin via inhibiting xanthine oxidase activity.

  4. Antioxidant effect of naturally occurring xanthines on the oxidative damage of DNA bases

    Science.gov (United States)

    Vieira, A. J. S. C.; Telo, J. P.; Pereira, H. F.; Patrocínio, P. F.; Dias, R. M. B.

    1999-01-01

    The repair of the oxidised radicals of adenine and guanosine by several naturally occurring xanthines was studied. Each pair of DNA purine/xanthine was made to react with the sulphate radical and the decrease of the concentration of both compounds was measured by HPLC as a function of irradiation time. The results show that xanthine efficiently prevents the oxidation of the two DNA purines. Theophyline and paraxanthine repair the oxidised radical of adenine but not the one from guanosine. Theobromine and caffeine do not show any protecting effect. An order of the oxidation potentials of all the purines studied is proposed. La réparation des radicaux oxydés de l'adénine et de la guanosine par des xanthines naturelles a été étudiée en soumettant chaque paire base de l'ADN/xanthine à l'oxydation par le radical sulfate et en mesurant par HPLC la disparition des deux composés en fonction du temps d'irradiation. Les résultats montrent que la xanthine joue un rôle protecteur efficace contre l'oxydation des deux purines de l'ADN. La théophyline et la paraxanthine réparent le radical oxydé de l'adénine mais pas celui de la guanosine. La théobromine et la cafeíne n'ont pas d'effet protecteur. Un ordre de potentiels d'oxydation des purines étudiées est proposé.

  5. Milk Allergy

    Science.gov (United States)

    ... processed meats and breakfast cereals. Hidden sources of milk include: Whey Casein Ingredients spelled with the prefix "lact" — such ... produced by using enzymes to break down (hydrolyze) milk proteins, such as casein or whey. Further processing can include heat and filtering. Depending ...

  6. Occupational asthma caused by inhalation of bovine serum albumin powder

    OpenAIRE

    Choi, Gil-Soon; Kim, Joo-Hee; Lee, Haet-Nim; Sung, Jun-Mo; Lee, Jin-Woo; Park, Hae-Sim

    2009-01-01

    Bovine serum albumin (BSA), which is present in bovine plasma, is one of the major allergens affecting patients with food allergies induced by milk and meat. It is also commonly used in research laboratories. Although some reports have documented food allergies associated with BSA, BSA-induced occupational asthma has not been reported. We report a case of occupational asthma and rhinitis in a laboratory worker caused by the inhalation of BSA powder, in which an IgE-mediated response was sugge...

  7. Which factors in raw cow's milk contribute to protection against allergies?

    Science.gov (United States)

    van Neerven, R J Joost; Knol, Edward F; Heck, Jeroen M L; Savelkoul, Huub F J

    2012-10-01

    Several epidemiologic studies have shown that growing up in a farming environment is associated with a decreased risk of allergies. A factor that correlates strongly with this effect is the early ingestion of unheated cow's milk. Although, to date, no controlled studies on raw milk consumption have been performed to formally demonstrate this effect, several factors in bovine milk have been described that might explain how raw cow's milk consumption can decrease the risk of allergies. In addition, increasing knowledge on the immunologically active factors in breast milk have also contributed to our understanding of the effects of bovine milk in infants because many of the factors in bovine milk are expected to have functional effects in human subjects as well. Here we review these factors and their mechanisms of action and compare their presence in bovine milk and breast milk. A better understanding of these factors, as well as how to retain them, might ultimately lead to the development of mildly processed milk and infant nutrition products that could become a part of preventive strategies to reduce the incidence of allergic disease.

  8. Genome-wide association and biological pathway analysis for milk-fat composition in Danish Holstein and Danish Jersey cattle

    DEFF Research Database (Denmark)

    Buitenhuis, Bart; Janss, Luc L G; Poulsen, Nina Aagaard;

    2014-01-01

    The milk fat profile of the Danish Holstein (DH) and Danish Jersey (DJ) show clear differences. Identification of the genomic regions, genes and biological pathways underlying the milk fat biosynthesis will improve the understanding of the biology underlying bovine milk fat production and may...... provide new possibilities to change the milk fat composition by selective breeding. In this study a genome wide association scan (GWAS) in the DH and DJ was performed for a detailed milk fatty acid (FA) profile using the HD bovine SNP array and subsequently a biological pathway analysis based on the SNP...

  9. Genetic polymorphisms in milk protein genes and their impact on milk composition.

    Science.gov (United States)

    Dovc, P

    2000-01-01

    About 40 different genetic variants are present at bovine milk protein loci and considerable differences in allele frequencies were observed among breeds. Advanced electrophoretic methods and recently, the DNA technology have been applied for identification and characterisation of milk protein genetic variants. Mutations with impact on gene expression have been identified. The influence of lactoprotein genetic variants on milk composition and cheese making ability has been studied extensively. The most prominent effects have been found for beta-LG A/B and kappa-CN A/B in cattle and alpha s1-CN E in goat.

  10. Improvement of the antimicrobial and antioxidant activities of camel and bovine whey proteins by limited proteolysis.

    Science.gov (United States)

    Salami, Maryam; Moosavi-Movahedi, Ali Akbar; Ehsani, Mohammad Reza; Yousefi, Reza; Haertlé, Thomas; Chobert, Jean-Marc; Razavi, Seyed Hadi; Henrich, Robert; Balalaie, Saeed; Ebadi, Seyed Ahmad; Pourtakdoost, Samineh; Niasari-Naslaji, Amir

    2010-03-24

    The compositions and structures of bovine and camel milk proteins are different, which define their functional and biological properties. The aim of this study was to investigate the effects of enzymatic hydrolysis of camel and bovine whey proteins (WPs) on their antioxidant and antimicrobial properties. After enzymatic treatment, both the antioxidant and the antimicrobial activities of bovine and camel WPs were improved. The significantly higher antioxidant activity of camel WPs and their hydrolysates as compared with that of bovine WPs and their hydrolysates may result from the differences in amounts and/or in accessibilities of antioxidant amino acid residues present in their primary structures and from the prevalence of alpha-lactalbumin and beta-lactoglobulin as proteolytic substrates in camel and bovine whey, respectively. The results of this study reveal differences in antimicrobial and antioxidant activities between WP hydrolysates of bovine and camel milk and the effects of limited proteolysis on these activities.

  11. Antibiotic Sensitivity Pattern of Staphylococcus aureus and Escherichia coli Isolated from Bovine Fresh Milk (POLA SENSITIVITAS ANTIBIOTIK TERHADAP STAPHYLOCOCCUS AUREUS DAN ESCHERICHIA COLI YANG DIISOLASI DARI SUSU SAPI SEGAR

    Directory of Open Access Journals (Sweden)

    Lucia Ratna Winata Muslimin

    2016-01-01

    Full Text Available The aims of this study were to determine the sensitivity of S.aureus and E. coli isolated fromfresh milk against against several antibiotics and to determine the safety of the milk for humancomsumsion. Milk was collected from milking diary cow and was used for the bacterial isolation. E.coli were were identified using Total Plate Count (TPC, Gram staining, their growth on Endo Agarand Eosin MethyleneBlue Agar, Biochemical analysis including glucose, lactose, sucrose,maltose, andsorbitol would be followed by Sorbitol Mac Conkey Agar Test for the identification of E.coliO157:H7.The isolation and identification of S.aureus were performed using Gram stain, TPC, growth on BairdParker Agar and Mannitol Salt Agar. S. aureus and S. epidermidis were differentiated by coagulaseand catalase tests. The antibiotic sensitivity tests for both S. aureus and E.coli were carried out usingthe following antibiotics: ampicillin, bacitracin, vancomycin, cefoperazone, ceftriaxone, cefotamine,cefuroxime, cefepime, cefazoline, ceftazidime, chloramphenicol, tetracycline, doxycycline, amikacin,kanamycin, neomycin, ertapenem, meropenem, imipenem, erythromycin, gentamycin, nalidixic acid,ciprofloxacin, levofloxacine, norfloxacine, ofloxacin, and novobiocin. Fresh milk obtained from thefarm was positive for S.aureus and E.coli and resistant to most antibiotics tested. The best antibioticsfor S. aureus were imipenem (54.1 mm, ampicillin (42.3 mm, cefazolin (41.6 mm, doxycycline (41.15mm, and for E.coli were Imipenem (30.1 mm, ertapenem (29.5 mm, and meropenem (25.35 mm. Thebovine fresh milk examined was contaminated by S.aureus and E.coli and to some extent, were alsoresistant to most antibiotics tested.

  12. Camel and bovine chymosin: the relationship between their structures and cheese-making properties

    Energy Technology Data Exchange (ETDEWEB)

    Langholm Jensen, Jesper [University of Copenhagen, (Denmark); Chr. Hansen A/S, Bøge Allé 10-12, DK-2970 Hørsholm (Denmark); Mølgaard, Anne; Navarro Poulsen, Jens-Christian [University of Copenhagen, (Denmark); Harboe, Marianne Kirsten [Chr. Hansen A/S, Bøge Allé 10-12, DK-2970 Hørsholm (Denmark); Simonsen, Jens Bæk [University of Copenhagen, (Denmark); Lorentzen, Andrea Maria; Hjernø, Karin [University of Southern Denmark, (Denmark); Brink, Johannes M. van den; Qvist, Karsten Bruun [Chr. Hansen A/S, Bøge Allé 10-12, DK-2970 Hørsholm (Denmark); Larsen, Sine, E-mail: sine@chem.ku.dk [University of Copenhagen, (Denmark)

    2013-05-01

    Analysis of the crystal structures of the two milk-clotting enzymes bovine and camel chymosin has revealed that the better milk-clotting activity towards bovine milk of camel chymosin compared with bovine chymosin is related to variations in their surface charges and their substrate-binding clefts. Bovine and camel chymosin are aspartic peptidases that are used industrially in cheese production. They cleave the Phe105-Met106 bond of the milk protein κ-casein, releasing its predominantly negatively charged C-terminus, which leads to the separation of the milk into curds and whey. Despite having 85% sequence identity, camel chymosin shows a 70% higher milk-clotting activity than bovine chymosin towards bovine milk. The activities, structures, thermal stabilities and glycosylation patterns of bovine and camel chymosin obtained by fermentation in Aspergillus niger have been examined. Different variants of the enzymes were isolated by hydrophobic interaction chromatography and showed variations in their glycosylation, N-terminal sequences and activities. Glycosylation at Asn291 and the loss of the first three residues of camel chymosin significantly decreased its activity. Thermal differential scanning calorimetry revealed a slightly higher thermal stability of camel chymosin compared with bovine chymosin. The crystal structure of a doubly glycosylated variant of camel chymosin was determined at a resolution of 1.6 Å and the crystal structure of unglycosylated bovine chymosin was redetermined at a slightly higher resolution (1.8 Å) than previously determined structures. Camel and bovine chymosin share the same overall fold, except for the antiparallel central β-sheet that connects the N-terminal and C-terminal domains. In bovine chymosin the N-terminus forms one of the strands which is lacking in camel chymosin. This difference leads to an increase in the flexibility of the relative orientation of the two domains in the camel enzyme. Variations in the amino acids

  13. Prevalence of coagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastitis

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Dangler, C. A.; Sordillo, L. M.

    1995-01-01

    This study was conducted to investigate polymorphism of the coagulase gene of Staphylococcus aureus causing bovine mastitis. One hundred eighty-seven strains of S. aureus were isolated from bovine mastitic milk samples obtained from 187 different Danish dairy farms. The isolates were characterised...

  14. Amperometric determination of xanthine in fish meat by zinc oxide nanoparticle/chitosan/multiwalled carbon nanotube/polyaniline composite film bound xanthine oxidase.

    Science.gov (United States)

    Devi, Rooma; Yadav, Sandeep; Pundir, C S

    2012-02-07

    Xanthine oxidase (XOD) was immobilized on a composite film of zinc oxide nanoparticle/chitosan/carboxylated multiwalled carbon nanotube/polyaniline (ZnO-NP/CHIT/c-MWCNT/PANI) electrodeposited over the surface of a platinum (Pt) electrode. A xanthine biosensor was fabricated using XOD/ZnO-NP/CHIT/c-MWCNT/PANI/Pt as working electrode, Ag/AgCl as reference electrode and Pt wire as auxiliary electrode connected through a potentiostat. The ZnO-NPs were characterized by X-ray diffraction (XRD) and transmission electron microscopy (TEM), and the enzyme electrode was characterized by cyclic voltammetry, scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and electrochemical impedance spectroscopy (EIS). The biosensor showed optimum response within 4 s at 0.5 V potential, pH 7.0, 35 °C and linear range 0.1-100 μM with a detection limit of 0.1 μM. The enzyme electrode was employed for determination of xanthine in fish meat during storage. The electrode lost 30% of its initial activity after 80 uses over one month, when stored at 4 °C.

  15. Detection of cow's milk proteins and minor components in human milk using proteomics techniques.

    Science.gov (United States)

    Coscia, A; Orrù, S; Di Nicola, P; Giuliani, F; Varalda, A; Peila, C; Fabris, C; Conti, A; Bertino, E

    2012-10-01

    Cow's milk proteins (CMPs) are the best characterized food allergens. The aim of this study was to investigate cow's milk allergens in human colostrum of term and preterm newborns' mothers, and other minor protein components by proteomics techniques, more sensitive than other techniques used in the past. Sixty-two term and 11 preterm colostrum samples were collected, subjected to a treatment able to increase the concentration of the most diluted proteins and simultaneously to reduce the concentration of the proteins present at high concentration (Proteominer Treatment), and subsequently subjected to the steps of proteomic techniques. The most relevant finding in this study was the detection of the intact bovine alpha-S1-casein in human colostrum, then bovine alpha-1-casein could be considered the cow's milk allergen that is readily secreted in human milk and could be a cause of sensitization to cow's milk in exclusively breastfed predisposed infants. Another interesting result was the detection, at very low concentrations, of proteins previously not described in human milk (galectin-7, the different isoforms of the 14-3-3 protein and the serum amyloid P-component), probably involved in the regulation of the normal cell growth, in the pro-apoptotic function and in the regulation of tissue homeostasis. Further investigations are needed to understand if these families of proteins have specific biological activity in human milk.

  16. Microorganism Utilization for Synthetic Milk

    Science.gov (United States)

    Morford, Megan A.; Khodadad, Christina L.; Caro, Janicce I.; Spencer, LaShelle E.; Richards, Jeffery T.; Strayer, Richard F.; Birmele, Michele N.; Wheeler, Raymond M.

    2014-01-01

    A desired architecture for long duration spaceflight, like aboard the International Space Station or for future missions to Mars, is to provide a supply of fresh food crops for the astronauts. However, some crops can create a high proportion of inedible plant waste. The main goal of the Synthetic Biology project, Cow in a Column, was to produce the components of milk (sugar, lipid, protein) from inedible plant waste by utilizing microorganisms (fungi, yeast, bacteria). Of particular interest was utilizing the valuable polysaccharide, cellulose, found in plant waste, to naturally fuel-through microorganism cellular metabolism- the creation of sugar (glucose), lipid (milk fat), and protein (casein) in order to produce a synthetic edible food product. Environmental conditions such as pH, temperature, carbon source, aeration, and choice microorganisms were optimized in the laboratory and the desired end-products, sugars and lipids, were analyzed. Trichoderma reesei, a known cellulolytic fungus, was utilized to drive the production of glucose, with the intent that the produced glucose would serve as the carbon source for milk fat production and be a substitute for the milk sugar lactose. Lipid production would be carried out by Rhodosporidium toruloides, yeast known to accumulate those lipids that are typically found in milk fat. Results showed that glucose and total lipid content were below what was expected during this phase of experimentation. In addition, individual analysis of six fatty acids revealed that the percentage of each fatty acid was lower than naturally produced bovine milk. Overall, this research indicates that microorganisms could be utilized to breakdown inedible solid waste to produce useable products. For future work, the production of the casein protein for milk would require the development of a genetically modified organism, which was beyond the scope of the original project. Additional trials would be needed to further refine the required

  17. Enzymatic Release and Characterization of Novel Bioactive Peptides from Milk Proteins

    DEFF Research Database (Denmark)

    De Gobba, Cristian

    Milk is considered the most complete food, providing most of the nutrients needed. Milk proteins are not only important for their function as a source of amino acids, but they are also a source of bioactive peptides. These are short amino acid sequences with different activities that have......-inhibitory, antioxidant and antimicrobial peptides) released from milk proteins by mean of enzyme-catalysed hydrolysis. Goat milk fractions (produced using microfiltration membranes) and bovine casein were used as substrates. The goat milk fractions (retentate, permeate and skimmed milk) were hydrolysed with two...... protein hydrolysates made in other studies. Regarding radical scavenging activity, the bovine casein hydrolysates also showed a positive correlation between extent of hydrolysis and activity, although the difference between the unhydrolysed sample and the hydrolysates was less marked. The goat milk...

  18. Chemical nature and reaction mechanisms of the molybdenum cofactor of xanthine oxidoreductase.

    Science.gov (United States)

    Okamoto, Ken; Kusano, Teruo; Nishino, Takeshi

    2013-01-01

    Xanthine oxidoreductase (XOR), a complex flavoprotein, catalyzes the metabolic reactions leading from hypoxanthine to xanthine and from xanthine to urate, and both reactions take place at the molybdenum cofactor. The enzyme is a target of drugs for therapy of gout or hyperuricemia. We review the chemical nature and reaction mechanisms of the molybdenum cofactor of XOR, focusing on molybdenum-dependent reactions of actual or potential medical importance, including nitric oxide (NO) synthesis. It is now generally accepted that XOR transfers the water-exchangeable -OH ligand of the molybdenum atom to the substrate. The hydroxyl group at OH-Mo(IV) can be replaced by urate, oxipurinol and FYX-051 derivatives and the structures of these complexes have been determined by xray crystallography under anaerobic conditions. Although formation of NO from nitrite or formation of xanthine from urate by XOR ischemically feasible, it is not yet clear whether these reactions have any physiological significance since the reactions are catalyzed at a slow rate even under anaerobic conditions.

  19. Extension model of lactation curves to evaluate the effect of the recombinant bovine somatotropin on milk yield in Holstein cows Modelo de extensão de curvas de lactação para avaliar o efeito da somatotropina bovina recombinante sobre a produção de leite em vacas Holstein

    Directory of Open Access Journals (Sweden)

    A. Palacios-Espinosa

    2010-02-01

    Full Text Available An extension model of lactation curves was used to determine the effect of recombinant bovine somatotropin (bST-r on milk yield in Holstein dairy cattle. This model use the fitted values obtained by the Wood model, and was tested on the records of 66 cows. The milk yield predicted with the extension model and the observed yield were compared and no significant differences were observed (P>0.05. Once the extension model was validated, the milk yield tests of 199 cows were used. The cows received bST-r 500mg by subcutaneous injections. The injections were applied after 100 days in milk at 14-day intervals (seven injections. The observed milk yield was compared with the yield expected by the extension model. An increase of 5.3% was observed in milk yield in response to the bST-r. This increase is lower than that reported in the literature in response to the growth hormone in dairy cattle. It is concluded that extension model used in the present work is reliable for extending the lactation curve in Holstein cows, and the increase in milk yield in response to the application of bST-r, determined in the same animal using the extension model, was lower than that reported by other authors.Um modelo de extensão de curvas de lactância foi utilizado para determinar o efeito da somatotropina bovina recombinante (bST-r sobre a produção de leite em vacas Holstein. Este modelo, que utiliza os valores ajustados obtidos pelo modelo de Wood, foi testato nos registros de 66 vacas. A produção de leite predita com o modelo de extensão e a produção observada foram comparadas e não se observaram diferenças significativas (P>0,05. Uma vez validado o modelo de extensão, utilizaram-se os controles de produção de leite (de cada 15 dias de 199 vacas. As vacas receberam injeções de 500mg de bST-r via subcutânea. As injeções fora aplicadas a partir dos 100 dias de lactação a intervalos de 14 dias (sete injeções. A produção de leite observada foi

  20. Transfer of dietary zinc and fat to milk--evaluation of milk fat quality, milk fat precursors, and mastitis indicators.

    Science.gov (United States)

    Wiking, L; Larsen, T; Sehested, J

    2008-04-01

    The present study demonstrated that the zinc concentration in bovine milk and blood plasma is significantly affected by the intake of saturated fat supplements. Sixteen Holstein cows were used in a 4 x 4 Latin square design with 4 periods of 12 d, and 4 dietary treatments were conducted. A total mixed ration based on corn silage, grass-clover silages, and pelleted sugar beet pulp was used on all treatments. A high de novo milk fat diet was formulated by adding rapeseed meal and molasses in the total mixed ration [39 mg of Zn/kg of dry matter (DM)], and a low de novo diet by adding saturated fat, fat-rich rapeseed cake, and corn (34 mg of Zn/kg of DM). Dietary Zn levels were increased by addition of ZnO to 83 and 80 mg of Zn/kg of DM. Treatments did not affect daily DM intake, or yield of energy-corrected milk, milk fat, or milk protein. The high de novo diet significantly increased milk fat percentage and milk content of fatty acids with chain length from C6 to C16, and decreased content of C18 and C18:1. Treatments did not influence milk free fatty acids at 4 degrees C at 0 or 28 h after milking. The average diameter of milk fat globules was significantly greater in milk from cows offered low de novo diets. Furthermore, the low de novo diet significantly increased the concentration of nonesterified fatty acids and d-beta-hydroxybutyrate in blood plasma, the latter was also increased in milk. Treatments did not affect the enzyme activity of lactate dehydrogenase and N-acetyl-beta-d-glucosaminidase in milk or the activity of isocitrate dehydrogenase and malate dehydrogenase in blood plasma. The low de novo diet significantly increased plasma Zn and milk Zn content, whereas dietary Zn level did not in itself influence these parameters. This indicates that the transfer of fat from diet to milk might facilitate transfer of Zn from diet to milk.

  1. Perspectives on Immunoglobulins in colostrum and milk

    DEFF Research Database (Denmark)

    Hurley, W L; Theil, Peter Kappel

    2011-01-01

    Immunoglobulins form an important component of the immunological activity found in milk and colostrum. They are central to the immunological link that occurs when the mother transfers passive immunity to the offspring. The mechanism of transfer varies among mammalian species. Cattle provide...... a readily available immune rich colostrum and milk in large quantities, making those secretions important potential sources of immune products that may benefit humans. Immune milk is a term used to describe a range of products of the bovine mammary gland that have been tested against several human diseases....... The use of colostrum or milk as a source of immunoglobulins, whether intended for the neonate of the species producing the secretion or for a different species, can be viewed in the context of the types of immunoglobulins in the secretion, the mechanisms by which the immunoglobulins are secreted...

  2. Inactivation of Escherichia coli glutamine synthetase by xanthine oxidase, nicotinate hydroxylase, horseradish peroxidase, or glucose oxidase: effects of ferredoxin, putidaredoxin, and menadione.

    Science.gov (United States)

    Stadtman, E R; Wittenberger, M E

    1985-06-01

    Previous studies have shown that several mixed-function oxidation (MFO) systems are capable of catalyzing the inactivation of glutamine synthetase (GS) [R.L. Levine, C. N. Oliver, R. M. Fulks, and E. R. Stadtman (1978) Proc. Natl. Acad. Sci. USA 78, 2120-2124] and a number of the other enzymes [L. Fucci, C. N. Oliver, M. J. Coon, and E. R. Stadtman (1983) Proc. Natl. Acad. Sci. USA 80, 1521-1525]. It has now been found that in the presence of Fe(III), O2, and an appropriate electron donor (hypoxanthine or NADPH, respectively) glutamine synthetase is also inactivated by either milk xanthine oxidase or Clostridial nicotinate hydroxylase. Inactivation of glutamine synthetase by either of these flavoproteins is greatly stimulated by the presence of electron carrier proteins possessing nonheme-iron-sulfur (NHIS) clusters (i.e., ferredoxin or putidaredoxin) or by the presence of menadione. The inactivation reactions are partially inhibited by free radical scavengers, superoxide dismutase, (SOD), histidine, mannitol, dimethyl sulfoxide, and dimethylthiourea, and are inhibited completely by either Mn(II), EDTA, or catalase. The sensitivity to SOD inhibition is greatly suppressed when the xanthine oxidase system is supplemented with either ferredoxin or redoxin. In the presence of the latter NHIS-proteins (and only when they are present), MFO systems, comprised of either horseradish peroxidase and H2O2 or glucose oxidase, O2, and glucose, can also catalyze the inactivation of GS. The ability of ferredoxin and putidaredoxin to promote oxidation modification of GS by any one of these MFO systems suggests that proteins with NHIS centers may mediate the generation (or stabilization) of highly reactive radical intermediates.

  3. 77 FR 29914 - Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products

    Science.gov (United States)

    2012-05-21

    ... RIN 0579-AC68 Bovine Spongiform Encephalopathy; Importation of Bovines and Bovine Products AGENCY... live bovines and products derived from bovines with regard to bovine spongiform encephalopathy. This... with regard to bovine spongiform encephalopathy. Comments on the proposed rule were required to......

  4. Milk Thistle (PDQ)

    Science.gov (United States)

    ... Professionals Questions to Ask about Your Treatment Research Milk Thistle (PDQ®)–Patient Version Overview Go to Health ... supplement (see Question 9 ). Questions and Answers About Milk Thistle What is milk thistle? Milk thistle is ...

  5. A peptidomic analysis of human milk digestion in the infant stomach reveals protein-specific degradation patterns.

    Science.gov (United States)

    Dallas, David C; Guerrero, Andrés; Khaldi, Nora; Borghese, Robyn; Bhandari, Aashish; Underwood, Mark A; Lebrilla, Carlito B; German, J Bruce; Barile, Daniela

    2014-06-01

    In vitro digestion of isolated milk proteins results in milk peptides with a variety of actions. However, it remains unclear to what degree protein degradation occurs in vivo in the infant stomach and whether peptides previously annotated for bioactivity are released. This study combined nanospray LC separation with time-of-flight mass spectrometry, comprehensive structural libraries, and informatics to analyze milk from 3 human mothers and the gastric aspirates from their 4- to 12-d-old postpartum infants. Milk from the mothers contained almost 200 distinct peptides, demonstrating enzymatic degradation of milk proteins beginning either during lactation or between milk collection and feeding. In the gastric samples, 649 milk peptides were identified, demonstrating that digestion continues in the infant stomach. Most peptides in both the intact milk and gastric samples were derived from β-casein. The numbers of peptides from β-casein, lactoferrin, α-lactalbumin, lactadherin, κ-casein, serum albumin, bile salt-associated lipase, and xanthine dehydrogenase/oxidase were significantly higher in the gastric samples than in the milk samples (P milk and gastric samples (P milk peptides with immunomodulatory and antibacterial properties of clinical relevance to the proximal intestinal tract. Data are available via ProteomeXchange (identifier PXD000688).

  6. Técnica rápida de neutralização viral para a detecção de anticorpos contra o vírus da Diarréia Viral Bovina (BVDV no leite A rapid virus-neutralization test for detection of antibodies against bovine viral diarrhea virus (BVDV in milk

    Directory of Open Access Journals (Sweden)

    Scherer Charles Fernando Capinos

    2002-04-01

    positivos através do leite enviado rotineiramente para contagem de células somáticas (CCS, reduzindo significativamente os custos com a coleta individual, transporte e teste de amostras.The identification of bovine viral diarrhea virus (BVDV positive herds through detection of antibodies in milk may viabilize large scale control/eradication programs. With this objective, the virus neutralization test (VN was adapted to detect BVDV antibodies in milk. The adaptation consisted of a reduction in the time of incubation followed by detection of viral antigens in the indicator cells by immunofluorescence (IFA and allowed readings at 24 hours. The rapid virus neutralization test (RVN was initially tested in 1,335 serum samples, showing a 93.7% sensitivity and 91.1% agreement with the traditional VN. The RVN was also used to test 423 bovine sera that were toxic for cell culture in the traditional VN test, detecting 316 (74.7% positive samples. Testing of matched serum and milk samples from BVDV seropositive cows showed that the VNR can detect antibodies in the milk of cows with serum neutralizing titers as low as 10. Anti-BVDV neutralizing activity was detected in milk of 97.4% (191/196 of cows with serum titers ³320; in 92.9% (79/85 of cows with titers of 160; in 88% (59/67 of cows with serum titers of 80. The frequency of BVDV antibodies in milk was 76.9% (40/52 for cows with serum titers of 40; 61.3% (19/31 for cows with titers of 20 and 33.3% (10/30 for cows with serum titers of 20. These results demonstrate that the RVN test is adequate for detecting BVDV antibodies in milk, mainly in cows having moderate to high serum titers, and therefore may be used for testing bulk milk samples to identify herds with viral activity. The use of this test may viabilize large scale programs for control/eradication of BVDV infection. It allows to assay a large number of samples and identify positive herds through testing milk routinely submitted for somatic cell counts (SCC, reducing costs

  7. Mediterranean milk and milk products.

    Science.gov (United States)

    Hinrichs, Jörg

    2004-03-01

    Milk and dairy products are part of a healthy Mediterranean diet which, besides cow's milk, also consists of sheep's, goat's and buffalo's milk--alone or as a mixture---as raw material. The fat and protein composition of the milk of the various animal species differs only slightly, but in every case it has a high priority in human nutrition. The milk proteins are characterized by a high content of essential amino acids. Beyond that macromolecules,which have various biological functions, are available or may be formed by proteolysis in milk. Taking this into consideration, the technology of different well-known Italian and German cheese types is presented and the differences as well as correspondences regarding nutrition are discussed. Especially Ricotta and Mascarpone are discussed in detail. Ricotta represents a special feature as this cheese is traditionally made of whey and cream. Thus the highly valuable whey proteins which contain a higher amount of the amino acids lysine, methionine and cysteic acid in comparison to casein and, additionally, to soy protein, are made usable for human nutrition. Finally, it is pointed out on the basis of individual examples that technologies to enrich whey proteins in cheese are already available and in use. Thus, the flavor of low fat cheese is improved and the nutritional value is increased.

  8. Mastitomics, the integrated omics of bovine milk in an experimental model of Streptococcus uberis mastitis: 2. Label-free relative quantitative proteomics† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6mb00290k Click here for additional data file.

    Science.gov (United States)

    Mudaliar, Manikhandan; Tassi, Riccardo; Thomas, Funmilola C.; McNeilly, Tom N.; Weidt, Stefan K.; McLaughlin, Mark; Wilson, David; Burchmore, Richard; Herzyk, Pawel; Eckersall, P. David

    2016-01-01

    Mastitis, inflammation of the mammary gland, is the most common and costly disease of dairy cattle in the western world. It is primarily caused by bacteria, with Streptococcus uberis as one of the most prevalent causative agents. To characterize the proteome during Streptococcus uberis mastitis, an experimentally induced model of intramammary infection was used. Milk whey samples obtained from 6 cows at 6 time points were processed using label-free relative quantitative proteomics. This proteomic analysis complements clinical, bacteriological and immunological studies as well as peptidomic and metabolomic analysis of the same challenge model. A total of 2552 non-redundant bovine peptides were identified, and from these, 570 bovine proteins were quantified. Hierarchical cluster analysis and principal component analysis showed clear clustering of results by stage of infection, with similarities between pre-infection and resolution stages (0 and 312 h post challenge), early infection stages (36 and 42 h post challenge) and late infection stages (57 and 81 h post challenge). Ingenuity pathway analysis identified upregulation of acute phase protein pathways over the course of infection, with dominance of different acute phase proteins at different time points based on differential expression analysis. Antimicrobial peptides, notably cathelicidins and peptidoglycan recognition protein, were upregulated at all time points post challenge and peaked at 57 h, which coincided with 10 000-fold decrease in average bacterial counts. The integration of clinical, bacteriological, immunological and quantitative proteomics and other-omic data provides a more detailed systems level view of the host response to mastitis than has been achieved previously. PMID:27412694

  9. Milk Intolerance, Beta-Casein and Lactose.

    Science.gov (United States)

    Pal, Sebely; Woodford, Keith; Kukuljan, Sonja; Ho, Suleen

    2015-08-31

    True lactose intolerance (symptoms stemming from lactose malabsorption) is less common than is widely perceived, and should be viewed as just one potential cause of cows' milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows' milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows' milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed.

  10. Milk Intolerance, Beta-Casein and Lactose

    Directory of Open Access Journals (Sweden)

    Sebely Pal

    2015-08-01

    Full Text Available True lactose intolerance (symptoms stemming from lactose malabsorption is less common than is widely perceived, and should be viewed as just one potential cause of cows’ milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows’ milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows’ milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed.

  11. Solvent Binding Analysis and Computational Alanine Scanning of the Bovine Chymosin-Bovine κ-Casein Complex Using Molecular Integral Equation Theory.

    Science.gov (United States)

    Palmer, David S; Sørensen, Jesper; Schiøtt, Birgit; Fedorov, Maxim V

    2013-12-10

    We demonstrate that the relative binding thermodynamics of single-point mutants of a model protein-peptide complex (the bovine chymosin-bovine κ-casein complex) can be calculated accurately and efficiently using molecular integral equation theory. The results are shown to be in good overall agreement with those obtained using implicit continuum solvation models. Unlike the implicit continuum models, however, molecular integral equation theory provides useful information about the distribution of solvent density. We find that experimentally observed water-binding sites on the surface of bovine chymosin can be identified quickly and accurately from the density distribution functions computed by molecular integral equation theory. The bovine chymosin-bovine κ-casein complex is of industrial interest because bovine chymosin is widely used to cleave bovine κ-casein and to initiate milk clotting in the manufacturing of processed dairy products. The results are interpreted in light of the recent discovery that camel chymosin is a more efficient clotting agent than bovine chymosin for bovine milk.

  12. In vitro xanthine oxidase inhibitory activity of methanol extracts of Erythrina indica Lam. leaves and stem bark

    Institute of Scientific and Technical Information of China (English)

    Kandhasamy Sowndhararajan; Jince Mary Joseph; Dharmar Rajendrakumaran

    2012-01-01

    Objective: To determine the total phenolic content and in vitro xanthine oxidase inhibitory activity of methanol extracts of leaves and stem bark of Erythrina indica. Methods: Folin-ciocalteu method was used to determine the total phenolic content. Xanthine oxidase inhibitory activity was assayed spectrophotometrically and the degree of enzyme inhibition was determined by measuring the increase in absorbance at 295nm associated with uric acid formation. Results:The methanol extract of stem bark of E. indica contains higher level of total phenolic content (412.8 mg GAE/g extract) and also exhibited higher xanthine oxidase inhibition activity (IC50 52.75μg/mL) than the leaves. Conclusions: It could be concluded that the stem bark of E. indica was highly effective in xanthine oxidase inhibition and might be used for the gout related disorders.

  13. Early gradual feeding with bovine colostrum improves gut function and NEC resistance relative to infant formula in preterm pigs

    DEFF Research Database (Denmark)

    Shen, René L.; Thymann, Thomas; Østergaard, Mette V.;

    2015-01-01

    It is unclear when and how to start enteral feeding for preterm infants when mother’s milk is not available. We hypothesized that early and slow advancement with either formula or bovine colostrum stimulates gut maturation and prevents necrotizing enterocolitis (NEC) in preterm pigs, used as models...... intestinal permeability, compared with BC pigs (all P Early feeding with formula induces intestinal dysfunction whereas bovine colostrum supports gut maturation when mother’s milk is absent during the first week after preterm birth...

  14. An overview of bovine α-lactalbumin structure and functionality

    Directory of Open Access Journals (Sweden)

    Nicoleta STĂNCIUC

    2010-12-01

    Full Text Available α-Lactalbumin is the second major protein in bovine milk (2-5% of the total protein in bovine milk. The human variant has several physiologic functions in the neonatal period. In the mammary gland, itparticipates in lactose synthesis and facilitates milk production and secretion. α-Lactalbumin binds divalent cations (Ca2+, Zn2+ and may facilitate the absorption of essential minerals. Also, it provides awell-balanced supply of essential amino acids for the growing infant. During its digestion, peptides with antibacterial and imunostimulatory properties are formed, thereby possibly helping in the protection against infection. The protein fractions enriched with α-lactalbumin may be added to infant formula to provide some of the benefits of this protein. This article reviews a number of studies which may contribute to a more coherent understanding of the structure and some functional properties of α-lactalbumin.

  15. Antibody Tracing, Seroepidemiology and Risk Factors of Bovine Respiratory Syncytial Virus and Bovine Adenovirus-3 in Dairy Holstein Farms

    Directory of Open Access Journals (Sweden)

    Mahsa FARZINPOUR

    2016-01-01

    Full Text Available Antibody tracing, risk factors and seroepidemiology of bovine respiratory syncytial virus and bovine adenovirus-3 were investigated in 22 Industrial and Semi-Industrial dairy Holstein farms. Serum samples (n=736 from various ages of unvaccinated cows were collected from May to September 2012. Risk factors including age, past history of respiratory diseases, amount of milk production, husbandry type and herd size were considered. Data were analyzed by Chi-square and logistic regression. Results indicated that the infection with some of individual viruses was related to past history of respiratory disease and herd size. No specific pattern was seen on the effect of level of milk production on seropositivity of animals. The seroprevalence for BRSV and BAV-3 were 89.1% and 88%, respectively. The present study indicates that infections of bovine respiratory viruses frequently occur in cattle of Fars province and the main viral cause of primary occurrence of respiratory diseases may be due to aforementioned viruses.

  16. Absorption and retention of free and milk protein-bound cyano- and hydroxocobalamins

    DEFF Research Database (Denmark)

    Kornerup, Linda Skibsted; Juul, Christian Bredgaard; Fedosov, Sergey

    2016-01-01

    alone or bound to milk protein. Materials and methods We synthesized labeled OH[57Co]Cbl from commercially available CN[57Co]Cbl. Recombinant bovine transcobalamin (rbTC) was produced in yeast and skimmed milk obtained off the shelf. Male Wistar rats (250–300 g) received labeled Cbl by gastric gavage...

  17. Genetic variation of milk fatty acid composition between and within dairy cattle breeds

    NARCIS (Netherlands)

    Maurice - Van Eijndhoven, M.H.T.

    2014-01-01

    Abstract Maurice – Van Eijndhoven, M.H.T. (2014). Genetic variation of milk fatty acid composition between and within dairy cattle breeds. PhD thesis, Wageningen University, the Netherlands Fat is one of the main components in bovine milk and comprises a large number of indivi

  18. Consumption of unprocessed cow's milk protects infants from common respiratory infections

    NARCIS (Netherlands)

    Loss, G.; Depner, M.; Ulfman, L.H.; Neerven, van R.J.J.; Hose, A.J.; Genuneit, J.

    2015-01-01

    Background: Breast-feeding is protective against respiratory infections in early life. Given the co-evolutionary adaptations of humans and cattle, bovine milk might exert similar anti-infective effects in human infants. Objective: To study effects of consumption of raw and processed cow's milk on co

  19. Comparison between genetic parameters of cheese yield and nutrient recovery or whey loss traits measured from individual model cheese-making methods or predicted from unprocessed bovine milk samples using Fourier-transform infrared spectroscopy.

    Science.gov (United States)

    Bittante, G; Ferragina, A; Cipolat-Gotet, C; Cecchinato, A

    2014-10-01

    Cheese yield is an important technological trait in the dairy industry. The aim of this study was to infer the genetic parameters of some cheese yield-related traits predicted using Fourier-transform infrared (FTIR) spectral analysis and compare the results with those obtained using an individual model cheese-producing procedure. A total of 1,264 model cheeses were produced using 1,500-mL milk samples collected from individual Brown Swiss cows, and individual measurements were taken for 10 traits: 3 cheese yield traits (fresh curd, curd total solids, and curd water as a percent of the weight of the processed milk), 4 milk nutrient recovery traits (fat, protein, total solids, and energy of the curd as a percent of the same nutrient in the processed milk), and 3 daily cheese production traits per cow (fresh curd, total solids, and water weight of the curd). Each unprocessed milk sample was analyzed using a MilkoScan FT6000 (Foss, Hillerød, Denmark) over the spectral range, from 5,000 to 900 wavenumber × cm(-1). The FTIR spectrum-based prediction models for the previously mentioned traits were developed using modified partial least-square regression. Cross-validation of the whole data set yielded coefficients of determination between the predicted and measured values in cross-validation of 0.65 to 0.95 for all traits, except for the recovery of fat (0.41). A 3-fold external validation was also used, in which the available data were partitioned into 2 subsets: a training set (one-third of the herds) and a testing set (two-thirds). The training set was used to develop calibration equations, whereas the testing subsets were used for external validation of the calibration equations and to estimate the heritabilities and genetic correlations of the measured and FTIR-predicted phenotypes. The coefficients of determination between the predicted and measured values in cross-validation results obtained from the training sets were very similar to those obtained from the whole

  20. Development, optimization and biological evaluation of chitosan scaffold formulations of new xanthine derivatives for treatment of type-2 diabetes mellitus.

    Science.gov (United States)

    Lupascu, Florentina Geanina; Dash, Mamoni; Samal, Sangram Keshari; Dubruel, Peter; Lupusoru, Catalina Elena; Lupusoru, Raoul-Vasile; Dragostin, Oana; Profire, Lenuta

    2015-09-18

    New xanthine derivatives as antidiabetic agents were synthesized and new chitosan formulations have been developed in order to improve their biological and pharmacokinetic profile. Their physicochemical properties in terms of particle size, morphology, swelling degree, crystalline state, the loading efficiency as well as in vitro release and biodegradation rate were evaluated. According to the results the optimized formulations have a high drug loading efficiency (more than 70%), small particle size, a good release profile in the simulated biological fluids (the percentage of cumulative release being more than 55%) and improved biodegradation rate in reference with chitosan microparticles. The presence of xanthine derivatives (6, 7) in chitosan microparticles was demonstrated by means of FTIR analysis. The X-ray diffraction (XRD) proved that xanthine derivatives present a crystalline state. The biological evaluation assays confirmed the antioxidant and antidiabetic effects of the xanthine derivatives (6, 7) and their chitosan formulations (CS-6, CS-7). Xanthine derivative 6 showed a high antiradical scavenging effect (DPPH remaining=41.78%). It also reduced the glucose blood level with 59.30% and recorded level of glycosylated hemoglobin was 4.53%. The effect of its chitosan formulation (CS-6) on the level of blood glucose (114.5mg/dl) was even more intense than the one recorded by pioglitazone (148.5mg/dl) when used as standard antidiabetic drug. These results demonstrated the potential application of xanthine derivative 6 and its chitosan formulation (CS-6) in the treatment of the diabetes mellitus syndrome.

  1. Disposable amperometric biosensors based on xanthine oxidase immobilized in the Prussian blue modified screen-printed three-electrode system

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The screen-printed three-electrode system was applied to fabricate a new type of disposable amperometric xanthine oxidase biosensor.Carbon-working,carbon-counter and Ag/AgCl reference electrodes were all manually printed on the polyethylene terephthalate substrate forming the screen-printed three-electrode system by the conventional screen-printing process.As a mediator,Prussian blue could not only catalyze the electrochemical reduction of hydrogen peroxide produced from the enzyme reaction,but also keep the favorable potential around 0 V.The optimum operational conditions,including pH,potential and temperature,were investigated.The sensitivities of xanthine and hypoxanthine detections were 13.83 mA/M and 25.56 mA/M,respectively.A linear relationship was obtained in the concentration range between 0.10μM and 4.98μM for xanthine and between 0.50μM and 3.98μM for hypoxanthine.The small Michaelis-menten constant value of the xanthine oxidase biosensor was calculated to be 3.90 μM.The results indicate that the fabricated xanthine oxidase biosensor is effective and sensitive for the detection of xanthine and hypoxanthine.

  2. Novel insights into the inhibitory mechanism of kaempferol on xanthine oxidase.

    Science.gov (United States)

    Wang, Yajie; Zhang, Guowen; Pan, Junhui; Gong, Deming

    2015-01-21

    Xanthine oxidase (XO), a key enzyme in purine catabolism, is widely distributed in human tissues. It can catalyze xanthine to generate uric acid and cause hyperuricemia and gout. Inhibition kinetics assay showed that kaempferol inhibited XO activity reversibly in a competitive manner. Strong fluorescence quenching and conformational changes of XO were found due to the formation of a kaempferol-XO complex, which was driven mainly by hydrophobic forces. The molecular docking further revealed that kaempferol inserted into the hydrophobic cavity of XO to interact with some amino acid residues. The main inhibition mechanism of kaempferol on XO activity may be due to the insertion of kaempferol into the active site of XO occupying the catalytic center of the enzyme to avoid the entrance of the substrate and inducing conformational changes of XO. In addition, luteolin exhibited a stronger synergistic effect with kaempferol than did morin at the lower concentration.

  3. Extraction optimization of polyphenols, antioxidant and xanthine oxidase inhibitory activities from Prunus salicina Lindl.

    Directory of Open Access Journals (Sweden)

    Yibin LI

    2016-01-01

    Full Text Available Abstract Optimization of polyphenols extraction from plum (Prunus salicina Lindl. was evaluated using response surface methodology. The Box-Behnken experimental results showed the optimal conditions involved an extraction temperature of 59 °C, a sonication time of 47 min, and an ethanol concentration of 61% respectively. The maximum extraction yield of total polyphenols was 44.74 mg gallic acid equivalents per gram of dried plum at optimal conditions. Polyphenol extracts exhibited stronger antioxidant activities than Vc by evaluating of 1,1-diphenyl-2-picrylhydrazyl (DPPH and hydroxyl radical scavenging activity. Furthermore, polyphenol extracts (IC50 = 179 g/mL showed obvious inhibitory effects on xanthine oxidase. These findings suggest that polyphenol extracts from P. salicina can be potentially used as natural antioxidant and xanthine oxidase inhibitory agents.

  4. Inhibitory effects of cardols and related compounds on superoxide anion generation by xanthine oxidase.

    Science.gov (United States)

    Masuoka, Noriyoshi; Nihei, Ken-ichi; Maeta, Ayami; Yamagiwa, Yoshiro; Kubo, Isao

    2015-01-01

    5-Pentadecatrienylresorcinol, isolated from cashew nuts and commonly known as cardol (C₁₅:₃), prevented the generation of superoxide radicals catalysed by xanthine oxidase without the inhibition of uric acid formation. The inhibition kinetics did not follow the Michelis-Menten equation, but instead followed the Hill equation. Cardol (C₁₀:₀) also inhibited superoxide anion generation, but resorcinol and cardol (C₅:₀) did not inhibit superoxide anion generation. The related compounds 3,5-dihydroxyphenyl alkanoates and alkyl 2,4-dihydroxybenzoates, had more than a C9 chain, cooperatively inhibited but alkyl 3,5-dihydroxybenzoates, regardless of their alkyl chain length, did not inhibit the superoxide anion generation. These results suggested that specific inhibitors for superoxide anion generation catalysed by xanthine oxidase