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Sample records for bovine milk xanthine

  1. Fluorescence quenching study of quercetin interaction with bovine milk xanthine oxidase

    NASA Astrophysics Data System (ADS)

    Rasoulzadeh, Farzaneh; Jabary, Hamideh Nadjarpour; Naseri, Abdolhossein; Rashidi, Mohammad-Reza

    2009-02-01

    Quercetin is a natural flavonoid with many important therapeutic properties. The interaction of this polyphenolic compound bovine milk xanthine oxidase as one of its major target proteins was studied using fluorescence quenching method for the first time. It was found that the fluorescence quenching of xanthine oxidase occurs through a static mechanism. The results revealed the presence of a single binding site on xanthine oxidase with the binding constant value equals to 1.153 × 10 4 l mol -1 at 310 K and pH 7.4. The thermodynamic parameters were also calculated at different temperatures. The enthalpy and entropy changes were found as -10.661 kJ mol -1 and +43.321 J mol -1 K -1 indicating that both hydrogen binding and hydrophobic are involved in the interaction of this polyphenolic natural compound with xanthine oxidase. The results may provide a ground for further studies with different flavonoids to find a safe alternative for allopurinol, the only xanthine oxidase inhibitor with clinical application.

  2. Preparation of monoclonal antibodies to xanthine oxidase and other proteins of bovine milk-fat-globule membrane.

    PubMed Central

    Mather, I H; Nace, C S; Johnson, V G; Goldsby, R A

    1980-01-01

    Nine hybridomas secreting monoclonal antibody to proteins of bovine milk-fat-globule membrane were isolated. All nine cell lines continued to secrete monoclonal antibody after serial transfer in culture and after passage as solid tumours in Balb/cJ mice. Four of the cell lines secreted monoclonal antibody specific for xanthine oxidase, one of the major proteins of milk-fat-globule membrane. PMID:6894088

  3. Posttranslational ruling of xanthine oxidase activity in bovine milk by its substrates

    SciTech Connect

    Silanikove, Nissim Shapiro, Fira; Leitner, Gabriel

    2007-11-23

    The aims of this study were to test the hypothesis that the substrates of xanthine oxidase (XO), xanthine and hypoxanthine, are consumed while the milk is stored in the gland between milkings, and to explore how XO activity responds to bacteria commonly associated with subclinical infections in the mammary gland. Freshly secreted milk was obtained following complete evacuation of the gland and induction of milk ejection with oxytocin. In bacteria-free fresh milk xanthine and hypoxanthine were converted to uric acid within 30 min (T{sub 1/2} {approx} 10 min), which in turn provides electrons for formation of hydrogen peroxide and endows the alveolar lumen with passive protection against invading bacteria. On the other hand, the longer residence time of milk in the cistern compartment was not associated with oxidative stress as a result of XO idleness caused by exhaustion of its physiological fuels. The specific response of XO to bacteria species and the resulting bacteria-dependent nitrosative stress further demonstrates that it is part of the gland immune system.

  4. Substrate specificity of guinea pig liver aldehyde oxidase and bovine milk xanthine oxidase for methyl- and nitrobenzaldehydes.

    PubMed

    Veskoukis, Aristidis S; Kouretas, Demetrios; Panoutsopoulos, Georgios I

    2006-01-01

    Both aldehyde oxidase and xanthine oxidase catalyze the oxidation of a wide range of N-heterocycles and aldehydes. These enzymes are important in the oxidation of N-heterocyclic xenobiotics, whereas their role in the oxidation of xenobiotic aldehydes is usually ignored. The present investigation describes the interaction of methyl- and nitrosubstituted benzaldehydes, in the ortho-, meta- and parapositions, with guinea pig liver aldehyde oxidase and bovine milk xanthine oxidase. The kinetic constants showed that most substituted benzaldehydes are excellent substrates of aldehyde oxidase with lower affinities for xanthine oxidase. Low Km values for aldehyde oxidase were observed with most benzaldehydes tested, with 3-nitrobenzaldehyde having the lowest Km value and 3-methylbenzaldehyde being the best substrate in terms of substrate efficiency (Ks). Additionally, low Km values for xanthine oxidase were found with most benzaldehydes tested. However, all benzaldehydes also had low Vmax values, which made them poor substrates of xanthine oxidase. It is therefore possible that aldehyde oxidase may be critical in the oxidation of xenobiotic and endobiotic derived aldehydes and its role in such reactions should not be ignored.

  5. Absorption of enzymatically active sup 125 I-labeled bovine milk xanthine oxidase fed to rabbits

    SciTech Connect

    Rzucidlo, S.J. ); Zikakis, J.P. )

    1990-05-01

    Rabbits fed a regular laboratory diet supplemented with a high-fat milk containing xanthine oxidase (XO) were studied to determine the presence of active XO in the blood. A pilot feeding study, where rabbits consumed a high-fat diet containing xanthine oxidase, showed a correlation between dairy food consumption and XO activity in the blood. Antibody to dietary XO was also found. In a second study, rabbits were fed ad libitum the high-fat milk and blood serum samples were tested weekly for XO activity. No elevation in serum XO activity was found. A third study showed that serum XO activity was increased when rabbits were force fed the high-fat milk. The final study consisted of force feeding {sup 125}I-labeled XO to one rabbit to ascertain whether the observed increase in serum XO was due to dietary or endogenous XO. Isoelectric focusing of sera collected from the test rabbit strongly suggested that at least a portion of the serum XO contained the radioactive label. This is the first direct evidence showing the uptake of dietary active XO from the gut.

  6. Purification and partial characterisation of camel milk xanthine oxidoreductase.

    PubMed

    Baghiani, A; Harrison, R; Benboubetra, M

    2003-12-01

    Xanthine oxidoreductase (XOR) was purified in the presence of dithiothrietol from camel milk with yields of up to 22.2mg/l that were comparable to those obtained from bovine and human milk sources. On SDS-PAGE, the freshly purified camel milk XOR had a protein flavin (A280/A450) ratio of 5.3 +/- 0.4 and appeared homogenous with a single major band of approximately Mr 145.3 KDa. Surprisingly, in all the batches (n = 8) purified camel milk XOR showed no detectable activity towards xanthine or NADH. The molybdenum content of camel XOR was comparable to human and goat milk enzymes. After resulphuration, camel milk XOR gave a specific activity of 1.1 nmol/min/mg and 13.0 nmol/min/mg enzyme towards pterin (fluorimetric assay) and xanthine (spectrophotometric assay) respectively. This activity was markedly lower than that of human, bovine and goat enzymes obtained under the same conditions. These findings suggest that the molybdo-form of camel enzyme is totally under desulpho inactive form. It is possible that camel neonates are equipped with an enzymic system that reactivates XOR in their gut and consequently generates antibacterial reactive oxygen species.

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

    PubMed

    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.

  8. A kinetic study of hypoxanthine oxidation by milk xanthine oxidase.

    PubMed Central

    Escribano, J; Garcia-Canovas, F; Garcia-Carmona, F

    1988-01-01

    The course of the reaction sequence hypoxanthine----xanthine----uric acid catalysed by xanthine:oxygen oxidoreductase from milk was investigated on the basis of u.v. spectra taken during the course of hypoxanthine and xanthine oxidations. It was found that xanthine accumulated in the reaction mixture when hypoxanthine was used as a substrate. The time course of the concentrations of hypoxanthine, xanthine intermediate and uric acid product was simulated numerically. The mathematical model takes into account the competition of substrate, intermediate and product and the accumulation of the intermediate at the enzyme. This type of analysis permits the kinetic parameters of the enzyme for hypoxanthine and xanthine to be obtained. PMID:3196295

  9. Bovine milk exosome proteome

    USDA-ARS?s Scientific Manuscript database

    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 ...

  10. Milk xanthine oxidase type D (dehydrogenase) and type O (oxidase). Purification, interconversion and some properties

    PubMed Central

    Battelli, Maria Giulia; Lorenzoni, E.; Stirpe, F.

    1973-01-01

    1. The xanthine oxidase of cow's milk, crude or purified, appears as an oxidase (type O), and can be converted almost completely into a NAD+-dependent dehydrogenase (type D) by treatment with dithioerythritol or dihydrolipoic acid, but only to a small extent by other thiols. 2. The D form of the enzyme is inhibited by NADH, which competes with NAD+. 3. The kinetic constants of the two forms of the enzyme are similar to those of the corresponding forms of rat liver xanthine oxidase. 4. Milk xanthine oxidase is converted into an irreversible O form by pretreatment with chymotrypsin, papain or subtilisin, but only partially with trypsin. 5. The enzyme as purified shows a major faster band and a minor slower band on gel electrophoresis. The slower band is greatly reinforced after xanthine oxidase is converted into the irreversible O form by chymotrypsin. ImagesFig. 5. PMID:4352904

  11. Is bovine milk a health hazard?

    PubMed

    Oski, F A

    1985-01-01

    Whole bovine milk should not be fed to infants during the first year of life because of its association with occult gastrointestinal bleeding, iron deficiency anemia, and cow's milk allergy. The consumption of whole milk after the first year of life should be discouraged because of its potential role in a variety of disorders including atherosclerosis, recurrent abdominal pain of childhood, cataracts, milk-borne infections, and juvenile delinquency.

  12. Invited review: Bovine milk fat globule membrane as a potential nutraceutical.

    PubMed

    Spitsberg, V L

    2005-07-01

    For the last 15 yr, a great deal of knowledge has been accumulated on health beneficial factors, protein and nonprotein, of bovine milk fat globule membrane (MFGM). Among the health-beneficial components of the MFGM are cholesterolemia-lowering factor, inhibitors of cancer cell growth, vitamin binders, inhibitor of Helicobacter pylori, inhibitor of beta-glucuronidase of the intestinal Escherichia coli, xanthine oxidase as a bactericidal agent, butyrophilin as a possible suppressor of multiple sclerosis, and phospholipids as agents against colon cancer, gastrointestinal pathogens, Alzheimer's disease, depression, and stress. All of the above compel us to consider bovine MFGM as a potential nutraceutical.

  13. Caries protective agents in human milk and bovine milk: an in vitro study.

    PubMed

    Shetty, Vabitha; Hegde, Amitha M; Nandan, S; Shetty, Suchetha

    2011-01-01

    To estimate Calcium and Phosphorus withdrawal from hydroxyapatite in the presence of bovine milk and human milk from which the following protective fractions namely Casein, Whey protein, Lactose and Milk fat have been individually removed and to compare the above protective fractions in human and bovine milk. Human milk obtained from lactating mothers in the labor ward of Kshema hospital was subjected to immediate analysis. Bovine milk was obtained from a local dairy. Equal quantities of human milk and bovine milk (1 ml) were separately subjected to the systematic removal of the four milk fractions. As each fraction was removed, the remaining milk samples were subjected to testing. Powdered hydroxyapatite from human dental enamel was subjected to demineralization with the addition of the milk sample under test for 20 minutes. This mixture was then centrifuged. Aliquots of the supernatant were taken for calcium and Phosphorus analysis using photospectrometry. Ten demineralization tests were similarly carried out for every milk fraction for both human and bovine milk separately. Equal samples of whole bovine milk and whole human milk were also subjected to similar testing. The calcium and phosphorus dissolution values were higher when the individual fractions were eliminated from both human milk/enamel samples and bovine milk/enamel samples as compared to the values obtained from whole human milk/whole bovine milk/enamel samples. Further higher calcium and phosphorus dissolution values were observed when the fractions were individually and separately removed from the whole human milk/enamel samples as compared to the corresponding values obtained when these fractions were removed from bovine milk/enamel samples. The evaluated milk fraction in bovine milk namely casein, whey protein, lactose and milk fat were individually more caries protective when compared to the corresponding fractions in human milk.

  14. Bioelectrical impedance analysis of bovine milk fat

    NASA Astrophysics Data System (ADS)

    Veiga, E. A.; Bertemes-Filho, P.

    2012-12-01

    Three samples of 250ml at home temperature of 20°C were obtained from whole, low fat and fat free bovine UHT milk. They were analysed by measuring both impedance spectra and dc conductivity in order to establish the relationship between samples related to fat content. An impedance measuring system was developed, which is based on digital oscilloscope, a current source and a FPGA. Data was measured by the oscilloscope in the frequency 1 kHz to 100 kHz. It was showed that there is approximately 7.9% difference in the conductivity between whole and low fat milk whereas 15.9% between low fat and free fat one. The change of fatness in the milk can be significantly sensed by both impedance spectra measurements and dc conductivity. This result might be useful for detecting fat content of milk in a very simple way and also may help the development of sensors for measuring milk quality, as for example the detection of mastitis.

  15. Structural and Functional Insights into the Catalytic Inactivity of the Major Fraction of Buffalo Milk Xanthine Oxidoreductase

    PubMed Central

    Gadave, Kaustubh S.; Panda, Santanu; Singh, Surender; Kalra, Shalini; Malakar, Dhruba; Mohanty, Ashok K.; Kaushik, Jai K.

    2014-01-01

    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 (kcat/Km) 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. PMID:24498153

  16. Cariogenicity and acidogenicity of human milk, plain and sweetened bovine milk: an in vitro study.

    PubMed

    Prabhakar, A R; Kurthukoti, Ameet J; Gupta, Pranjali

    2010-01-01

    The objective of the present study was to determine the acidogenicity and cariogenicity of human breast milk and plain and sweetened packaged bovine milk. First all milk specimens were inoculated with a cariogenic strain of Streptococcus mutans (SM). The culture pH and number of colony forming units (cfus) was assessed. Second, the buffer capacity of all milk specimens was evaluated by mixing with acid. Finally, enamel windows were created on extracted primary maxillary incisors and colonized with SM. Enamel demineralization and caries progression were assessed visually, histologically, and radiographically at the end of twelve weeks. Plain and sweetened packaged bovine milk (BM) supported greater bacterial growth and caused more fermentation than human breast milk (HBM). The buffer capacity values for plain and sweetened bovine milk were highest; HBM, however had poor buffering capacity. The progression of the carious lesions into the dentin was most severe for the sweetened bovine milk. HBM and plain bovine milk are relatively cariogenic in an in vitro caries model in the absence of saliva. However, supplementation with sugar exponentially enhances the cariogenic potential of the natural milk.

  17. Acidogenic potential of soy and bovine milk beverages.

    PubMed

    Dashper, S G; Saion, B N; Stacey, M A; Manton, D J; Cochrane, N J; Stanton, D P; Yuan, Y; Reynolds, E C

    2012-09-01

    Soy beverages are water extracts of whole soybeans and are often promoted as a healthy alternative to bovine milk. Little analysis has been carried out to determine the effects of soy beverages on oral health, especially their potential acidogenicity. The aim of this study was to determine the potential acidogenicity of a range of soy and bovine milk beverages. In vitro acid production by Streptococcus mutans was measured in soy and milk beverages at a constant pH of 6.5 or 5.5, as was the fall in pH over a 10 min period. The acid buffering capacity and calcium and phosphate concentrations (total and soluble) of the beverages were also determined. The rate of acid production by S. mutans in the milk beverages was five to six times lower at pH 6.5 than in the soy beverages and three to five times lower at pH 5.5. Whilst the pH fall in the presence of S. mutans over 10 min was negligible in the milk beverages there was a significant decrease in pH in the soy beverages. This was also reflected in the lower buffering capacity of the soy beverages. The levels of soluble calcium in the soy beverages were lower than those in the milk beverages although total calcium contents were similar. Soy beverages have a higher potential acidogenicity than bovine milk beverages. Patients consider soy beverages to be a healthy, low cariogenic alternative to other beverages, including bovine milk. This study shows that soy beverages have a higher potential acidogenicity than bovine milk and therefore may have a greater potential cariogenicity. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. Prevalence of Listeria monocytogenes in raw bovine milk and milk products from central highlands of Ethiopia.

    PubMed

    Seyoum, Eyasu Tigabu; Woldetsadik, Daniel Asrat; Mekonen, Tesfu Kassa; Gezahegn, Haile Alemayehu; Gebreyes, Wondwossen Abebe

    2015-11-30

    Listeria monocytogenes is of major significance in human and veterinary medicine. Most human Listeria infections are foodborne and the association of contaminated milk and dairy produce consumption with human listeriosis is noteworthy. In Ethiopia, there is limited data regarding the prevalence of L. monocytogenes in raw bovine milk and dairy products. The aim of this study was, therefore, to determine the prevalence of L. monocytogenes in raw bovine milk and dairy produce. A total of 443 milk and milk product samples were microbiologically analyzed following methods recommended by the U.S. Food and Drug Administration Bacteriological Analytical Manual to isolate Listeria spp. The overall prevalence of Listeria spp. was 28.4% and specifically that of L. monocytogenes was 5.6%. Taking the prevalence of Listeria spp. into consideration, cheese was found to be highly contaminated at 60%, followed by pasteurized milk samples (40%), raw milk (18.9%) and yoghurt (5%). Considering the prevalence of Listeria monocytogenes only, raw milk had the lowest contamination while cheese had the highest, followed by pasteurized milk and yoghurt. Raw milk and milk products produced in urban and peri-urban areas of central Ethiopia were contaminated with pathogenic bacteria, L. monocytogenes. The detection of this pathogen in raw milk and milk products warrants an urgent regulatory mechanism to be put in place and also the potential role of milk processing plants in the contamination of dairy products should be investigated.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27999311

  20. Bioelectrical impedance analysis for bovine milk: Preliminary results

    NASA Astrophysics Data System (ADS)

    Bertemes-Filho, P.; Valicheski, R.; Pereira, R. M.; Paterno, A. S.

    2010-04-01

    This work reports the investigation and analysis of bovine milk quality by using biological impedance measurements using electrical impedance spectroscopy (EIS). The samples were distinguished by a first chemical analysis using Fourier transform midinfrared spectroscopy (FTIR) and flow citometry. A set of milk samples (100ml each) obtained from 17 different cows in lactation with and without mastitis were analyzed with the proposed technique using EIS. The samples were adulterated by adding distilled water and hydrogen peroxide in a controlled manner. FTIR spectroscopy and flow cytometry were performed, and impedance measurements were made in a frequency range from 500Hz up to 1MHz with an implemented EIS system. The system's phase shift was compensated by measuring saline solutions. It was possible to show that the results obtained with the Bioelectrical Impedance Analysis (BIA) technique may detect changes in the milk caused by mastitis and the presence of water and hydrogen peroxide in the bovine milk.

  1. Identification of inflammatory cells in bovine milk by flow cytometry.

    PubMed

    Redelman, D; Butler, S; Robison, J; Garner, D

    1988-09-01

    Cells recovered from normal or mastitic bovine milk were examined by flow cytometry. All milk samples contained particulate material that was heterogeneous in size and that produced a right-angle light-scatter signal equal to or greater than that produced by human or bovine neutrophils. Although this material labeled with Hoechst 33342, it produced fluorescence intensities below that of intact bovine cells, suggesting that it consisted of cell fragments. Mastitic milk additionally contained other populations of cells that were poorly resolved from the normal particulate material by size (electronic volume sensor) and right-angle light scatter. In order to improve this resolution, the milk cells were incubated with carboxydimethylfluorescein diacetate (CMFDA) to label intact cells. When milk samples labeled with CMFDA were examined by dual-parameter analysis using green fluorescence and right-angle light scatter, five or more populations of cells could be identified in mastitic milk. These populations included intact and degenerate neutrophils, lymphocytes, including both small and activated cells, monocytes, and large activated macrophages containing many vacuoles and phagocytosed particles. Using this procedure, all the animals in the University of Nevada-Reno Holstein dairy herd were tested once a month for 6 months. In addition, individual animals with mastitis were examined one or more times each day during the course of the inflammatory process. In the routine screening, the flow cytometric examination detected mastitis before overt symptoms developed. In cows identified to have mastitis, the flow cytometric examination provided prognostic information regarding the success of treatments.

  2. Transcriptional profiling of bovine milk using RNA sequencing

    PubMed Central

    2012-01-01

    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 molecular functions

  3. Meat and milk compositions of bovine clones.

    PubMed

    Tian, X Cindy; Kubota, Chikara; Sakashita, Kunihito; Izaike, Yoshiaki; Okano, Ryoichi; Tabara, Norio; Curchoe, Carol; Jacob, Lavina; Zhang, Yuqin; Smith, Sadie; Bormann, Charles; Xu, Jie; Sato, Masumi; Andrew, Sheila; Yang, Xiangzhong

    2005-05-03

    The technology is now available for commercial cloning of farm animals for food production, but is the food safe for consumers? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction. The cloned animals and the comparators were managed under the same conditions and received the same diet. The composition of the meat and milk from the clones were largely not statistically different from those of matched comparators, and all parameters examined were within the normal industry standards or previously reported values. The data generated from our match-controlled experiments provide science-based information desired by regulatory agencies to address public concerns about the safety of meat and milk from somatic animal clones.

  4. Meat and milk compositions of bovine clones

    PubMed Central

    Tian, X. Cindy; Kubota, Chikara; Sakashita, Kunihito; Izaike, Yoshiaki; Okano, Ryoichi; Tabara, Norio; Curchoe, Carol; Jacob, Lavina; Zhang, Yuqin; Smith, Sadie; Bormann, Charles; Xu, Jie; Sato, Masumi; Andrew, Sheila; Yang, Xiangzhong

    2005-01-01

    The technology is now available for commercial cloning of farm animals for food production, but is the food safe for consumers? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction. The cloned animals and the comparators were managed under the same conditions and received the same diet. The composition of the meat and milk from the clones were largely not statistically different from those of matched comparators, and all parameters examined were within the normal industry standards or previously reported values. The data generated from our match-controlled experiments provide science-based information desired by regulatory agencies to address public concerns about the safety of meat and milk from somatic animal clones. PMID:15829585

  5. Expanding the bovine milk proteome through extensive fractionation.

    PubMed

    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

  6. The bovine lactation genome: Insights into the evolution of mammalian milk

    USDA-ARS?s Scientific Manuscript database

    The newly assembled Bos Taurus genome sequence enables the linkage of bovine milk and lactation data with other mammalian genomes. Using publicly available milk proteome data and mammary expressed sequence tags, 197 milk protein genes and over 6,000 mammary genes were identified in the bovine genome...

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

    PubMed

    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.

  8. Short communication: characterization of soluble thiols in bovine milk.

    PubMed

    Niero, G; De Marchi, M; Masi, A; Penasa, M; Cassandro, M

    2015-09-01

    Antioxidants are molecules essential for the maintenance of cell homeostasis and their intake through the diet has positive effects on human health. Among antioxidants, low-molecular-weight (LMW) thiols represent an important class of compounds. The aim of this study was to identify LMW thiols in bovine milk. A total of 96 individual milk samples from Brown Swiss, Holstein-Friesian, Alpine Grey, and Simmental cattle breeds were collected in 8 herds. The LMW thiols were extracted from the soluble fraction of milk and, following a derivatization protocol, they were separated by reverse phase HPLC and detected fluorimetrically. Six thiol species were detected and 2, glutathione (GSH) and cysteine-glycine (Cys-Gly), were identified and quantified. Regardless of the breed, the average concentration of Cys-Gly in milk was greater than that of GSH. Overall, milk from dual-purpose breeds (Simmental and Alpine Grey) was richer in LMW thiols than milk from dairy cows (Holstein-Friesian and Brown Swiss). Glutathione and Cys-Gly, closely linked metabolically, were strongly correlated. Pearson correlations of Cys-Gly with protein and casein contents were moderately low, and no relationship was found between GSH and milk chemical composition. Future research should focus on the identification of all detected LMW thiol species.

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

    PubMed

    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. Physicochemical and acid gelation properties of commercial UHT-treated plant-based milk substitutes and lactose free bovine milk.

    PubMed

    Mäkinen, Outi E; Uniacke-Lowe, Thérèse; O'Mahony, James A; Arendt, Elke K

    2015-02-01

    Physicochemical and acid gelation properties of UHT-treated commercial soy, oat, quinoa, rice and lactose-free bovine milks were studied. The separation profiles were determined using a LUMiSizer dispersion analyser. Soy, rice and quinoa milks formed both cream and sediment layers, while oat milk sedimented but did not cream. Bovine milk was very stable to separation while all plant milks separated at varying rates; rice and oat milks being the most unstable products. Particle sizes in plant-based milk substitutes, expressed as volume mean diameters (d4.3), ranged from 0.55μm (soy) to 2.08μm (quinoa) while the average size in bovine milk was 0.52μm. Particles of plant-based milk substitutes were significantly more polydisperse compared to those of bovine milk. Upon acidification with glucono-δ-lactone (GDL), bovine, soy and quinoa milks formed structured gels with maximum storage moduli of 262, 187 and 105Pa, respectively while oat and rice milks did not gel. In addition to soy products currently on the market, quinoa may have potential in dairy-type food applications. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Mycoplasmacidal activity of bovine milk for T-mycoplasmas.

    PubMed

    Brownlie, J; Howard, C J; Gourlay, R N

    1974-12-01

    Normal bovine milk and whey was mycoplasmacidal for 6 of the 13 strains of bovine T-mycoplasmas examined. The in vitro assay used also demonstrated no killing of the human, canine and simian T-mycoplasma strains after 4 hr. incubation. However, there appeared to be some cow-to-cow variation in possession of this activity, and following E. coli endotoxin stimulation of the mammary gland the activity was considerably reduced.Whey from three normal cows was fractionated on a Bio-Gel A 1.5 m. column and the mycoplasmacidal activity of the resulting five peaks assayed. Only the second peak, peak B, contained activity and was characterized as the only peak containing bovine IgA. The active component in whey, however, was found to be heat stable at 60 degrees C. for 60 minutes and to pass through a dialysis membrane. This is inconsistent with it being immunoglobulin.

  12. Bovine somatotropin supplementation of dairy cows. Is the milk safe?

    PubMed

    Daughaday, W H; Barbano, D M

    Complex, biologically active proteins (eg, enzymes and hormones) can be manufactured safely and cost-effectively through applications of biotechnology. Some of these proteins (eg, human insulin, human somatotropin, rennet for cheese manufacture) are currently approved for medical or food processing applications. Bovine somatotropin (bST) for lactating dairy cattle is another product that can be produced via biotechnology and may allow dairy farmers to produce milk at a lower cost. In 1985, based on an evaluation of toxicological data, the Food and Drug Administration concluded that milk and meat from bST-supplemented cows was safe and wholesome. The Food and Drug Administration has authorized the use of milk and meat from bST-supplemented cows in the commercial food supply. Its evaluation of the impact of bST supplementation on the long-term health of dairy cattle is near completion, and bST may be approved for commercial use in early 1991.

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

    PubMed

    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

  14. Comparison of the principal proteins in bovine, caprine, buffalo, equine and camel milk.

    PubMed

    Hinz, Katharina; O'Connor, Paula M; Huppertz, Thom; Ross, R Paul; Kelly, Alan L

    2012-05-01

    Proteomic analysis of bovine, caprine, buffalo, equine and camel milk highlighted significant interspecies differences. Camel milk was found to be devoid of β-lactoglobulin, whereas β-lactoglobulin was the major whey protein in bovine, buffalo, caprine, and equine milk. Five different isoforms of κ-casein were found in camel milk, analogous to the micro-heterogeneity observed for bovine κ-casein. Several spots observed in 2D-electrophoretograms of milk of all species could tentatively be identified as polypeptides arising from the enzymatic hydrolysis of caseins. The understanding gained from the proteomic comparison of these milks may be of relevance both in terms of identifying sources of hypoallergenic alternatives to bovine milk and detection of adulteration of milk samples and products.

  15. Detecting β-Casein Variation in Bovine Milk.

    PubMed

    Caroli, Anna Maria; Savino, Salvatore; Bulgari, Omar; Monti, Eugenio

    2016-01-25

    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.

  16. Quantitative determination of complex carbohydrates in bovine milk and in milk-based infant formulas.

    PubMed

    Neeser, J R; Golliard, M; Del Vedovo, S

    1991-09-01

    Quantitative determination of all structural families of complex carbohydrate micronutrients was performed on bovine milk samples, milk-based infant formulas, and whey-based manufacturing raw materials. Differences found between formulas depended mainly on their whey: casein ratios. A solvent separation procedure was required for quantitative estimation of the gangliosides and neutral glycolipids within the fat fraction. All infant formulas except one contained slightly more gangliosides than bovine milk. Complex carbohydrates were consistently higher in the nonfat fraction. By gel permeation chromatography, an oligosaccharide subfraction was separated from a glycopeptide one. Oligosaccharide content of infant formulas increased as a function of the whey:casein ratio, and glycopeptides were found only in formulas made with whey components. Neuraminic acids from infant formulas were associated primarily with the glycoprotein fraction, except in hydrolysate-based preparations in which "precipitable" glycoproteins were converted into "soluble" glycopeptides by trypsin treatment. Because whey-based raw materials are very rich in all bovine milk glycoconjugates and oligosaccharides their increased use will result in high contents of these micronutrients in modern formulas.

  17. Effect of Prototheca zopfii on neutrophil function from bovine milk.

    PubMed

    Cunha, Luciane T; Pugine, Silvana P; Valle, Claudia R; Ribeiro, Andrea R; Costa, Ernane J X; De Melo, Mariza P

    2006-12-01

    This study was carried to investigate neutrophil function in the presence of Prototheca zopfii. For this purpose, bovine milk neutrophils were incubated in the absence (control) of and presence of P. zopfii, and then they were examined hydrogen peroxide (H(2)O(2)) production, antioxidant enzyme activities, and phagocytic capacity. Milk was collected from negative "California Mastitis Test" (CMT) quarter from three lactating Holstein cows after induction of leukocytosis with an intramammary infusion of oyster glycogen. H(2)O(2) production was measured using the phenol red method. Catalase activity was measured following H(2)O(2) reduction at 240 nm and the activity of glutathione reductase was determined by measuring the rate of NADPH oxidation at 340 nm. P. zopfii death was assessed by fluorescent microscopy using acridine orange assay and by colony forming units (CFUs). Comparisons between the groups were initially performed by analysis of variance (ANOVA). Significant differences were then compared using Tukey's test with a significance coefficient of 0.05. Hydrogen peroxide production, catalase and glutathione reductase activities by neutrophils incubated in presence of P. zopfii were stimulated five times, 21% and 27% respectively, compared to the unstimulated-neutrophils. Neutrophils did not affect P. zopfii death as shown by microscopy and CFUs. These observations led to the conclusion that the P. zopfii promote a high increase of H(2)O(2) production by neutrophils from bovine milk during algae exposition accompanied by increase of antioxidant enzyme activities; however, this process did not affect P. zopfii death.

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

    PubMed

    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

  19. An exclusively human milk-based diet is associated with a lower rate of necrotizing enterocolitis than a diet of human milk and bovine milk-based products.

    PubMed

    Sullivan, Sandra; Schanler, Richard J; Kim, Jae H; Patel, Aloka L; Trawöger, Rudolf; Kiechl-Kohlendorfer, Ursula; Chan, Gary M; Blanco, Cynthia L; Abrams, Steven; Cotten, C Michael; Laroia, Nirupama; Ehrenkranz, Richard A; Dudell, Golde; Cristofalo, Elizabeth A; Meier, Paula; Lee, Martin L; Rechtman, David J; Lucas, Alan

    2010-04-01

    To evaluate the health benefits of an exclusively human milk-based diet compared with a diet of both human milk and bovine milk-based products in extremely premature infants. Infants fed their own mothers' milk were randomized to 1 of 3 study groups. Groups HM100 and HM40 received pasteurized donor human milk-based human milk fortifier when the enteral intake was 100 and 40 mL/kg/d, respectively, and both groups received pasteurized donor human milk if no mother's milk was available. Group BOV received bovine milk-based human milk fortifier when the enteral intake was 100 mL/kg/d and preterm formula if no mother's milk was available. Outcomes included duration of parenteral nutrition, morbidity, and growth. The 3 groups (total n = 207 infants) had similar baseline demographic variables, duration of parenteral nutrition, rates of late-onset sepsis, and growth. The groups receiving an exclusively human milk diet had significantly lower rates of necrotizing enterocolitis (NEC; P = .02) and NEC requiring surgical intervention (P = .007). For extremely premature infants, an exclusively human milk-based diet is associated with significantly lower rates of NEC and surgical NEC when compared with a mother's milk-based diet that also includes bovine milk-based products. Copyright 2010 AUR. Published by Mosby, Inc. All rights reserved.

  20. Short communication: interaction of bovine milk protein with chlorpyrifos.

    PubMed

    Lv, Ying; Li, Xuefen; Wang, Zongyi; Zheng, Han; Zhang, Qi; Huo, Ran; Chen, Xiangning; Han, Tao

    2014-01-01

    Dairy products are considered as nutrient-dense foods and consumed by many people in western countries, as well as an increasing number of Asian people. Excessive and frequent application of pesticides on vegetables and fruits leads to a potential health hazard to consumers. The organophosphate insecticide chlorpyrifos has been reported to bind with human and bovine serum albumin. Thus, it is necessary to explore the interaction between food protein and chlorpyrifos. In this study, equilibrium dialysis and fluorescence spectra were used to demonstrate binding of milk proteins to chlorpyrifos. The amount of milk protein bound was 0.03±0.01mg/g. Moreover, the milk protein-chlorpyrifos complexes were stable at pH 3.5to 9.5 and ion concentrations from 0.1 to 1.0M. The amount of chlorpyrifos bound to milk proteins decreased to 50% after being in vitro digested by pepsin and trypsin. The results showed that the interaction between food proteins and the pesticide might partially remove the insecticide and reduce the concentration of pesticide absorbed into the blood and, thus, alleviate the corresponding toxicity.

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

    PubMed

    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.

  2. Bovine milk-derived exosomes for drug delivery

    PubMed Central

    Gupta, Ramesh C.

    2015-01-01

    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. PMID:26604130

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

    PubMed

    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.

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

    PubMed Central

    Magistrelli, Damiano; Rosi, Fabia

    2014-01-01

    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. PMID:28234329

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

    PubMed

    Magistrelli, Damiano; Rosi, Fabia

    2014-07-23

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

    PubMed

    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.

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

    PubMed Central

    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. PMID:26134630

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

    PubMed

    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.

  9. Purification of Golgi casein kinase from bovine milk.

    PubMed

    Duncan, J S; Wilkinson, M C; Burgoyne, R D

    2000-09-01

    Caseins and many other secretory proteins are phosphorylated during their transport through the secretory pathway by a protein kinase present within Golgi compartments. Molecular analysis of the Golgi casein kinase (GCK) has not been possible since it has not been purified to homogeneity or been cloned. Previous attempts have been made to purify GCK activity from mammary gland Golgi fractions, but these have not resulted in extensive purification of the enzyme. In the present study, we have demonstrated that substantial amounts of GCK activity, assayed using a specific peptide substrate, can be detected as a soluble form in bovine milk, and we have used milk as a source for purification. A purification protocol was established that allowed>80000-fold purification to a specific activity of GCK (approx. 700 nmoles/min per mg of protein) far higher than previously achieved. These findings cast doubts on previous claims for purification of GCK activity. In addition, ion-exchange chromatography resolved two closely eluting peaks of activity, suggesting the existence of two related, but distinct, GCK activities.

  10. Structure of glycopeptides isolated from bovine milk component PP3.

    PubMed

    Girardet, J M; Coddeville, B; Plancke, Y; Strecker, G; Campagna, S; Spik, G; Linden, G

    1995-12-15

    The heat-stable acid-soluble phosphoglycoprotein component PP3 was isolated from the bovine milk proteose peptone fraction by concanavalin A affinity chromatography. Glycopeptides were released by pronase digestion of the milk component PP3 and were subsequently separated by high-pH anion-exchange chromatography on CarboPac PA-1. The primary structures of the glycan and peptide moieties of eight N-glycopeptides have been established by combining methylation analysis, mass spectrometry, 400-MHz 1H-NMR spectroscopy, and peptide sequence analysis. All the analyzed fractions contained biantennary N-acetyllactosamine-type carbohydrate chains, some of them with a GalNAc(beta 1-4)GlcNAc or a NeuAc(alpha 2-6)GalNAc(beta 1-4)GlcNAc group. This particular sequence did or did not replace the Gal(beta 1-4)GlcNAc group usually found in most N-linked glycans. Moreover, the sialylated Gal and GalNAc residues were only found on the Man(alpha 1-3) antenna.

  11. Bovine papillomavirus DNA in milk, blood, urine, semen, and spermatozoa of bovine papillomavirus-infected animals.

    PubMed

    Lindsey, C L; Almeida, M E; Vicari, C F; Carvalho, C; Yaguiu, A; Freitas, A C; Beçak, W; Stocco, R C

    2009-01-01

    Papillomavirus infection in bovines is associated with cutaneous papillomatosis on the hide, udders and other epithelial tissues, as well as in oral respiratory, alimentary and urinary tract mucosa. Bovine papillomavirus (BPV) is also considered the etiological agent of esophageal tumors and the malignant bladder tumors that characterize the clinical condition associated with chronic enzootic hematuria. After infective viral DNA was found in cattle blood and BPV1, 2 and 4 DNA in cattle reproductive and embryonic tissues, we looked for and found BPV DNA in blood, milk, urine, seminal fluid, and spermatozoa of BPV-infected animals. Peripheral blood lymphocyte cultures from BPV-infected animals had high rates of chromosome aberrations, including radial rearrangements that signal oncogenic potential and viral interaction with telomeric regions. The finding of BPV DNA in body fluids and tissues other than the epithelium demonstrates co-infection of other tissues or cell types by papillomavirus and shows the potential role of lymphocytes, seminal fluid and spermatozoa in BPV transmission. Our findings reinforce a peremptory need for prophylactic and therapeutic instruments to curtail this disease in bovine livestock.

  12. Lipidomics profiling of goat milk, soymilk and bovine milk by UPLC-Q-Exactive Orbitrap Mass Spectrometry.

    PubMed

    Li, Qiangqiang; Zhao, Yan; Zhu, Dan; Pang, Xiumei; Liu, Yue; Frew, Russell; Chen, Gang

    2017-06-01

    Lipids are very important for human health and milk is a rich dietary source of lipids. In this study, the lipid content in three types of milk (goat, soy and bovine) were determined by using UPLC-Q-Exactive Orbitrap Mass Spectrometry. A total of 13 classes of lipids (including Cer, SM, LPC, PC, PE, DG, TG, PA, PG, PI, PS, LPE, FA) were measured. Moreover, lipid profiles differed significantly between the different milk types. Soymilk is rich in phospholipids including PC, PE, PS, PG, while goat milk is rich in medium chain triglycerides (MCT), USFA, ω-6 FA and ω-3 FA, especially EPA and DHA. Furthermore, a PLS model was established for differentiation of milk types based on the lipid profiles. A total of 14 lipids were identified as biomarkers for differentiation of milk types, thus providing a basis for milk authentication and detection of adulteration.

  13. Milk and fat production in dairy cattle influenced by advanced subclinical bovine leukemia virus infection.

    PubMed

    Wu, M C; Shanks, R D; Lewin, H A

    1989-02-01

    Genetic potentials (pedigree-estimated breeding value) for milk and for fat were compared in cows grouped according to subclinical stage of bovine leukemia virus infection. Genetic potential for milk production was significantly greater in seropositive cows with persistent lymphocytosis (622 +/- 72 kg) and in seropositive hematologically normal cows (554 +/- 34 34 kg) than in seronegative herdmates (418 +/- 53 kg). When 305-day twice-daily-milking mature equivalent milk production records for the current lactation were adjusted for genetic potential, bovine leukemia virus-infected cows that were hematologically normal had significantly greater milk production than did seronegative herdmates, suggesting that early bovine leukemia virus infection was positively associated with milk yield. Genetic potential for fat production was significantly greater for cows with persistent lymphocytosis (21 +/- 2 kg) than for other seropositive (16 +/- 1 kg) and seronegative herdmates (13 +/- 2 kg); however, 305-day twice-daily-milking mature equivalent fat production for the current lactation was not significantly different between the groups. Thus, cows with persistent lymphocytosis did not produce fat according to their genetic potential. As an apparent consequence of tendencies for greater milk yield and less fat production, milk fat percentage was significantly reduced in cows with persistent lymphocytosis (3.33 +/- 0.09%) and other seropositive cows (3.48 +/- 0.05%) relative to seronegative herdmates (3.67 +/- 0.07%). These results suggest a need to reevaluate the economic impact of bovine leukemia virus infection on the dairy industry.

  14. Milk and fat production in dairy cattle influenced by advanced subclinical bovine leukemia virus infection.

    PubMed Central

    Wu, M C; Shanks, R D; Lewin, H A

    1989-01-01

    Genetic potentials (pedigree-estimated breeding value) for milk and for fat were compared in cows grouped according to subclinical stage of bovine leukemia virus infection. Genetic potential for milk production was significantly greater in seropositive cows with persistent lymphocytosis (622 +/- 72 kg) and in seropositive hematologically normal cows (554 +/- 34 34 kg) than in seronegative herdmates (418 +/- 53 kg). When 305-day twice-daily-milking mature equivalent milk production records for the current lactation were adjusted for genetic potential, bovine leukemia virus-infected cows that were hematologically normal had significantly greater milk production than did seronegative herdmates, suggesting that early bovine leukemia virus infection was positively associated with milk yield. Genetic potential for fat production was significantly greater for cows with persistent lymphocytosis (21 +/- 2 kg) than for other seropositive (16 +/- 1 kg) and seronegative herdmates (13 +/- 2 kg); however, 305-day twice-daily-milking mature equivalent fat production for the current lactation was not significantly different between the groups. Thus, cows with persistent lymphocytosis did not produce fat according to their genetic potential. As an apparent consequence of tendencies for greater milk yield and less fat production, milk fat percentage was significantly reduced in cows with persistent lymphocytosis (3.33 +/- 0.09%) and other seropositive cows (3.48 +/- 0.05%) relative to seronegative herdmates (3.67 +/- 0.07%). These results suggest a need to reevaluate the economic impact of bovine leukemia virus infection on the dairy industry. PMID:2536940

  15. Cariogenicity of different commercially available bovine milk types in a biofilm caries model.

    PubMed

    Giacaman, Rodrigo A; Muñoz-Sandoval, Cecilia

    2014-01-01

    This study's purpose was to assess the cariogenicity of commercial bovine milk types in an experimental biofilm/caries model. Enamel and dentin slabs were used to grow biofilms of Streptococcus mutans UA159. Slabs/biofilms were exposed three times per day to commercial skim, semi-skim, whole, whole lactose-free, and whole with 10 percent sucrose-added bovine milk and to 10 percent sucrose and 0.9 percent sodium chloride as positive and negative caries-control, respectively. Biofilms were analyzed for bacterial counts, biomass, proteins, and polysaccharide production. Slab's demineralization was assessed by loss of surface microhardness and the biofilm acidogenicity by medium pH. Only whole and whole lactose-free milk kept pH above the demineralization threshold, inducing the lowest demineralization in both enamel and dentin (P<.05). Skim and semi-skim milk induced similar demineralization to the sucrose control, albeit slightly lower for semi-skim milk (P<.05). Whole and whole lactose-free milk produced lower biomass and less insoluble polysaccharides than the other treatments in enamel and dentin (P<.05). Adding 10 percent sucrose to whole milk turned it as cariogenic as 10 percent sucrose solution. Bovine whole milk seemed less cariogenic than sucrose and the other commercial milk types, but not anticariogenic. Fat content in milk seemed to reduce cariogenicity of the fluid.

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

    PubMed Central

    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

  17. [Effect of proteins from bovine milk serum on the multiplication of human cancerous cells].

    PubMed

    Bourtourault, M; Buléon, R; Sampérez, S; Jouan, P

    1991-01-01

    The addition of bovine milk whey to the culture medium of human cancerous cells (MCF-7 and PC-3) results in a significant reduction of cells growth. Milk whey acts more efficiently on MCF-7 than on PC-3 growth. The inhibition could be due to a protein. Its identification is now on progress.

  18. Human Milk Protein Production in Xenografts of Genetically Engineered Bovine Mammary Epithelial Stem Cells

    PubMed Central

    Martignani, Eugenio; Eirew, Peter; Accornero, Paolo

    2010-01-01

    Background In the bovine species milk production is well known to correlate with mammary tissue mass. However, most advances in optimizing milk production relied on improvements of breeding and husbandry practices. A better understanding of the cells that generate bovine mammary tissue could facilitate important advances in milk production and have global economic impact. With this possibility in mind, we show that a mammary stem cell population can be functionally identified and isolated from the bovine mammary gland. We also demonstrate that this stem cell population may be a promising target for manipulating the composition of cow's milk using gene transfer. Methods and Findings We show that the in vitro colony-forming cell assay for detecting normal primitive bipotent and lineage-restricted human mammary clonogenic progenitors are applicable to bovine mammary cells. Similarly, the ability of normal human mammary stem cells to regenerate functional bilayered structures in collagen gels placed under the kidney capsule of immunodeficient mice is shared by a subset of bovine mammary cells that lack aldehyde dehydrogenase activity. We also find that this activity is a distinguishing feature of luminal-restricted bovine progenitors. The regenerated structures recapitulate the organization of bovine mammary tissue, and milk could be readily detected in these structures when they were assessed by immunohistochemical analysis. Transplantation of the bovine cells transduced with a lentivirus encoding human β-CASEIN led to expression of the transgene and secretion of the product by their progeny regenerated in vivo. Conclusions These findings point to a common developmental hierarchy shared by human and bovine mammary glands, providing strong evidence of common mechanisms regulating the maintenance and differentiation of mammary stem cells from both species. These results highlight the potential of novel engineering and transplant strategies for a variety of commercial

  19. Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk.

    PubMed

    Kot, B; Piechota, M; Wolska, K M; Frankowska, A; Zdunek, E; Binek, T; Kłopotowska, E; Antosiewicz, M

    2012-01-01

    The aim of this study was to examine phenotypic and genotypic antimicrobial resistance of staphylococci from milk samples from cows with subclinical and clinical mastitis and from cows without mastitis symptoms to methicillin, tetracyclines, macrolides and lincosamides (ML). Of 207 strains, including 34 S. aureus and 173 coagulase-negative staphylococci (CNS), 11 (6.4%) CNS strains were phenotypically resistant to methicillin. The mecA gene was detected by PCR only in two S. xylosus strains and one strain of S. epidermidis and S. simulans. No methicillin-resistant S. aureus strains were observed. In methicillin-resistant strains with mecA, gene resistance to other investigated antibiotics was not observed. Phenotypic resistance to tetracycline was detected in 11.0% of CNS strains and 47.4% of them carried the tetK gene. Of 173 CNS strains studied, 27 (15.6%) were resistant to at least one ML antibiotic. The resistance gene ermC was detected in 55.5% of the 27 ML-resistant strains. The ermA and ermB genes were detected in 14.8% and 11.1% of ML-resistant CNS strains, respectively. Antimicrobial resistance to methicillin, tetracyclines and macrolides was detected more frequently in staphylococcal strains from clinical mastitis compared to animals with subclinical symptoms and without mastitis, while the resistance to lincosamides showed a similar frequency in all groups of cows. In conclusion, CNS species from bovine milk differ in phenotypic and genotypic antimicrobial resistance profiles, and the use of PCR technique alone for the detection of methicillin, macrolide, lincosamide and tetyracycline resistance in CNS from cattle is not reliable.

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

    PubMed

    Yamada, Tetsuya; Shigemura, Hiroaki; Ishiguro, Naotaka; Inoshima, Yasuo

    2013-01-01

    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.

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

    PubMed

    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.

  2. Inflammation-related microRNA expression level in the bovine milk is affected by mastitis

    PubMed Central

    Lai, Yu-Chang; Fujikawa, Takuro; Maemura, Tadashi; Ando, Takaaki; Kitahara, Go; Endo, Yasuyuki; Yamato, Osamu; Koiwa, Masateru; Kubota, Chikara

    2017-01-01

    MicroRNA (miRNA) in tissue and liquid samples have been shown to be associated with many diseases including inflammation. We aimed to identify inflammation-related miRNA expression level in the bovine mastitis milk. Expression level of inflammation-related miRNA in milk from mastitis-affected and normal cows was analyzed using qPCR. We found that expression level of miR-21, miR-146a, miR-155, miR-222, and miR-383 was significantly upregulated in California mastitis test positive (CMT+) milk. We further analyzed these miRNA using a chip-based QuantStudio Digital PCR System. The digital PCR results correlated with those of qPCR, demonstrating upregulation of miR-21, miR-146a, miR-155, miR-222, and miR-383 in CMT+ milk. In conclusion, we identified miRNA that are upregulated in CMT+ milk. These miRNA exhibited sensitivity and specificity greater than 80% for differentiating between CMT+ milk and normal milk. Our findings suggest that inflammation-related miRNA expression level in the bovine milk was affected by mastitis, and miRNA in milk have potential for use as biomarkers of bovine mastitis. PMID:28520748

  3. Inflammation-related microRNA expression level in the bovine milk is affected by mastitis.

    PubMed

    Lai, Yu-Chang; Fujikawa, Takuro; Maemura, Tadashi; Ando, Takaaki; Kitahara, Go; Endo, Yasuyuki; Yamato, Osamu; Koiwa, Masateru; Kubota, Chikara; Miura, Naoki

    2017-01-01

    MicroRNA (miRNA) in tissue and liquid samples have been shown to be associated with many diseases including inflammation. We aimed to identify inflammation-related miRNA expression level in the bovine mastitis milk. Expression level of inflammation-related miRNA in milk from mastitis-affected and normal cows was analyzed using qPCR. We found that expression level of miR-21, miR-146a, miR-155, miR-222, and miR-383 was significantly upregulated in California mastitis test positive (CMT+) milk. We further analyzed these miRNA using a chip-based QuantStudio Digital PCR System. The digital PCR results correlated with those of qPCR, demonstrating upregulation of miR-21, miR-146a, miR-155, miR-222, and miR-383 in CMT+ milk. In conclusion, we identified miRNA that are upregulated in CMT+ milk. These miRNA exhibited sensitivity and specificity greater than 80% for differentiating between CMT+ milk and normal milk. Our findings suggest that inflammation-related miRNA expression level in the bovine milk was affected by mastitis, and miRNA in milk have potential for use as biomarkers of bovine mastitis.

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

    PubMed

    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.

  5. The effect of heat treatment and skimming on precipitate formation in caprine and bovine milks.

    PubMed

    Miloradovic, Zorana N; Kljajevic, Nemanja V; Jovanovic, Snezana T; Vucic, Tanja R; Macej, Ognjen D

    2015-02-01

    Caprine and bovine milks have a similar overall gross composition, but vary considerably in the ratios of their casein components. These differences in colloidal casein micelles could affect directly or indirectly the heat stability of caprine and bovine milks at their natural pH. In the present work, the differences in colloidal stability of caprine and bovine milk have been studied by analysing the effect of heat treatment and skimming on precipitation of proteins. Raw and heated milk samples (70 °C/5 min, 80°C/5 min and 90°C/5 min) were centrifuged at 600, 2000, and 4500  g . The amount of precipitate formed after skimming was measured and the protein composition of both precipitates and supernatants analysed using the SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) and densitometry. In caprine milk, the heat treatment prior to skimming had a statistically significant effect on protein precipitation. Centrifugal force had a statistically significant effect on amount of precipitate for both milks, but the amount was 2 to 4 times higher for caprine milk. When defatting the milk for electrophoresis, a centrifugal force of 600  g appeared to be the most appropriate, in order to avoid protein loss and a possible error in the interpretation of results. Results of this study could also serve as the basis for further investigations on adjusting the skimming conditions for caprine milk in industrial dairy processing environment.

  6. Natural variability in bovine milk oligosaccharides from Danish Jersey and Holstein-Friesian breeds

    PubMed Central

    Sundekilde, Ulrik K; Barile, Daniela; Meyrand, Mickael; Poulsen, Nina A; Larsen, Lotte B; Lebrilla, Carlito B.; Bruce, German J.; Bertram, Hanne C

    2012-01-01

    Free oligosaccharides are key components of human milk and play multiple roles in the health of the neonate, by stimulating growth of selected beneficial bacteria in the gut, participating in development of the brain and exerting anti-pathogenic activity. However, the concentration of oligosaccharides is low in mature bovine milk, normally used for infant formula, compared with both human colostrum and mature human milk. Characterization of bovine milk oligosaccharides in different breeds is crucial for the identification of viable sources for oligosaccharide purification. An improved source of oligosaccharides can lead to infant formula with improved oligosaccharide functionality. In the present study we have analyzed milk oligosaccharides by high-performance liquid chromatography chip quadrupole time-of-flight mass spectrometry and performed a detailed data analysis using both univariate and multivariate methods. Both statistical tools revealed several differences in oligosaccharide profiles between milk samples from the two Danish breeds; Jersey and Holstein-Friesians. Jersey milk contained higher relative amounts of both sialylated and the more complex neutral fucosylated oligosaccharides, while the Holstein-Friesian milk had higher abundance of smaller and simpler neutral oligosaccharides. The statistical analyses revealed that Jersey milk contain significantly higher levels of fucosylated oligosaccharides than Holstein-Friesian milk. Jersey milk also possesses oligosaccharides with a higher degree of complexity and functional residues (fucose and sialic acid) suggesting it may therefore offer advantages in term of a wider array of bioactivities. PMID:22632419

  7. Supplementation of Infant Formula with Bovine Milk Fat Globule Membranes.

    PubMed

    Timby, Niklas; Domellöf, Magnus; Lönnerdal, Bo; Hernell, Olle

    2017-03-01

    Studies have shown that supplementation of infant formula with bovine milk fat globule membranes (MFGMs) may substantially narrow the gap in health outcomes between formula-fed and breastfed infants. In one study, consumption of a formula supplemented with a lipid-rich MFGM concentrate between 2 and 6 mo of age improved cognitive performance at 24 wk of age. In another study, a formula supplemented with a protein-rich MFGM concentrate given between 2 and 6 mo of age improved cognitive performance at 12 mo of age, decreased infectious morbidity until 6 mo of age, and yielded serum cholesterol concentrations closer to those of breastfed infants. A third study that assessed the safety of supplementing infant formula with a lipid-rich or a protein-rich MFGM concentrate found a noninferior weight gain for both groups compared with a nonsupplemented formula. In this study, there was an increased risk of eczema in the protein-rich group, but no serious adverse events. Infant formulas with supplemental MFGMs have been launched on the market in several countries. However, the evidence base must still be considered quite limited. Based on 3 randomized controlled trials that are not comparable, the intervention seems safe, but there is not enough evidence for a general recommendation on which MFGM fraction to use and at what concentration as formula supplement for a given outcome. © 2017 American Society for Nutrition.

  8. Transcriptome Profiling of Bovine Milk Oligosaccharide Metabolism Genes Using RNA-Sequencing

    PubMed Central

    Wickramasinghe, Saumya; Hua, Serenus; Rincon, Gonzalo; Islas-Trejo, Alma; German, J. Bruce; Lebrilla, Carlito B.; Medrano, Juan F.

    2011-01-01

    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 genes will guide

  9. Hydrolysis of bovine and caprine milk fat globules by lipoprotein lipase. Effects of heparin and skim milk on lipase distribution and on lipolysis

    SciTech Connect

    Sundheim, G.; Bengtsson-Olivecrona, G.

    1987-12-01

    Heparin can dissociate lipoprotein lipase from casein micelles, and addition of heparin enhances lipolysis in bovine but not in caprine milk. Heparin shortened the lag-time for binding of lipoprotein lipase to milk fat globules and for lipolysis. Heparin counteracted the inhibitory effects of skim milk on binding of lipase and on lipolysis. Heparin stimulated lipolysis in all bovine milk samples when added before cooling and in spontaneously lipolytic milk samples also when added after cooling. Heparin enhanced lipolysis of isolated milk fat globules. Hence, its effect is not solely due to dissociation of lipoprotein lipase from the casein micelles. Cooling of goat milk caused more marked changes in the distribution of lipase than cooling of bovine milk; the fraction of added /sup 125/I-labeled lipase that bound to cream increased from about 8 to 60%. In addition, caprine skim milk caused less inhibition of lipolysis than bovine skim milk. These observations provide an explanation for the high degree of cold storage lipolysis in goat milk. Heparin had only small effects on the distribution of lipoprotein lipase in caprine milk, which explains why heparin has so little effect on lipolysis in caprine milk. The distribution of /sup 35/S-labeled heparin in bovine milk was studied. In warm milk less than 10% bound to the cream fraction, but when milk was cooled, binding of heparin to cream increased to 45%. These results suggest that there exists in the skim fraction a relatively small amount of a heparin-binding protein, which on cooling of milk adsorbs to the milk fat, or suggests that cooling induces a conformational change in a membrane protein such that its affinity for heparin increases.

  10. Bovine Milk as a Source of Functional Oligosaccharides for Improving Human Health12

    PubMed Central

    Zivkovic, Angela M.; Barile, Daniela

    2011-01-01

    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. PMID:22332060

  11. Bovine milk in human nutrition – a review

    PubMed Central

    Haug, Anna; Høstmark, Arne T; Harstad, Odd M

    2007-01-01

    Milk and milk products are nutritious food items containing numerous essential nutrients, but in the western societies the consumption of milk has decreased partly due to claimed negative health effects. The content of oleic acid, conjugated linoleic acid, omega-3 fatty acids, short- and medium chain fatty acids, vitamins, minerals and bioactive compounds may promote positive health effects. Full-fat milk has been shown to increase the mean gastric emptying time compared to half-skimmed milk, thereby increasing the gastrointestinal transit time. Also the low pH in fermented milk may delay the gastric emptying. Hence, it may be suggested that ingesting full-fat milk or fermented milk might be favourable for glycaemic (and appetite?) regulation. For some persons milk proteins, fat and milk sugar may be of health concern. The interaction between carbohydrates (both natural milk sugar and added sugar) and protein in milk exposed to heat may give products, whose effects on health should be further studied, and the increasing use of sweetened milk products should be questioned. The concentration in milk of several nutrients can be manipulated through feeding regimes. There is no evidence that moderate intake of milk fat gives increased risk of diseases. PMID:17894873

  12. Bovine milk in human nutrition--a review.

    PubMed

    Haug, Anna; Høstmark, Arne T; Harstad, Odd M

    2007-09-25

    Milk and milk products are nutritious food items containing numerous essential nutrients, but in the western societies the consumption of milk has decreased partly due to claimed negative health effects. The content of oleic acid, conjugated linoleic acid, omega-3 fatty acids, short- and medium chain fatty acids, vitamins, minerals and bioactive compounds may promote positive health effects. Full-fat milk has been shown to increase the mean gastric emptying time compared to half-skimmed milk, thereby increasing the gastrointestinal transit time. Also the low pH in fermented milk may delay the gastric emptying. Hence, it may be suggested that ingesting full-fat milk or fermented milk might be favourable for glycaemic (and appetite?) regulation. For some persons milk proteins, fat and milk sugar may be of health concern. The interaction between carbohydrates (both natural milk sugar and added sugar) and protein in milk exposed to heat may give products, whose effects on health should be further studied, and the increasing use of sweetened milk products should be questioned. The concentration in milk of several nutrients can be manipulated through feeding regimes. There is no evidence that moderate intake of milk fat gives increased risk of diseases.

  13. Comparing water, bovine milk, and indoor residual spraying as possible sources of DDT and pyrethroid residues in breast milk.

    PubMed

    Sereda, Barbara; Bouwman, Henk; Kylin, Henrik

    2009-01-01

    The presence of pollutants in human breast milk is of major concern, especially in malaria control areas where 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) is currently used as indoor residual spray (IRS). The levels of DDT and pyrethroids (PYR) were determined in breast milk, bovine milk, and drinking water from northern KwaZulu-Natal, South Africa. Both reference and exposed mothers used the same market food, but the DDT levels in the exposed mothers (mean SigmaDDT 10 microg/g milk fat [mf]) were much higher than for the reference mothers (mean SigmaDDT 1.3 microg/g milk fat). This difference in residue levels indicates uptake from IRS-applied DDT, most likely via air and skin contact, and excludes food as the main source of pollutants. DDT levels in bovine milk (mean SigmaDDT 0.15 microg/g mf) from the exposed area were less than levels in breast milk from the reference area, and lower than the 20 microg/L maximum residue limit (MRL) set by the Food and Agriculture Organization (FAO). Mean SigmaDDT in water was 0.0065 microg/L, much lower then the WHO limit of the sum of all metabolites in drinking water of 1 microg/L, and therefore highly unlikely to have contributed to any extent toward levels in breast milk. Permethrin in breast milk (mean 1.1-1.6 microg/g milk fat) was probably derived from home garden and indoor use, while the other PYR (cypermethrin and cyfluthrin) at lower concentrations were probably derived from food and agricultural exposure. It is postulated that a better understanding of the indoor dynamics of DDT and other insecticides, through a concept of Total Homestead Environment Approach (THEA), is crucial for investigating options of reducing human exposure and uptake under malaria control conditions.

  14. Kappacin, a Novel Antibacterial Peptide from Bovine Milk

    PubMed Central

    Malkoski, Marina; Dashper, Stuart G.; O'Brien-Simpson, Neil M.; Talbo, Gert H.; Macris, Mary; Cross, Keith J.; Reynolds, Eric C.

    2001-01-01

    Caseinomacropeptide (CMP) is a heterogeneous C-terminal fragment (residues 106 to 169) of bovine milk κ-casein composed of glycosylated and phosphorylated forms of different genetic variants. We have demonstrated that CMP has growth-inhibitory activity against the oral opportunistic pathogens Streptococcus mutans and Porphyromonas gingivalis and against Escherichia coli. CMP was fractionated using reversed-phase high-performance liquid chromatography (RP-HPLC), and each fraction was tested for activity against S. mutans in a 96-well-plate broth assay. Fractions were characterized by N-terminal sequence analysis and mass spectrometry. The active form of CMP was shown to be the nonglycosylated, phosphorylated κ-casein (residues 106 to 169) [κ-casein(106–169)], which we have designated kappacin. Endoproteinase Glu-C was used to hydrolyze CMP, and the generated peptides were separated using RP-HPLC and gel filtration-HPLC and then tested for activity against S. mutans. The peptide Ser(P)149κ-casein-A(138–158) was the only peptide generated by endoproteinase Glu-C digestion that exhibited growth-inhibitory activity. Peptides corresponding to the sequences of the inhibitory peptide Ser(P)149κ-casein-A(138–158) and its nonphosphorylated counterpart κ-casein-A(138–158) were chemically synthesized and tested for antibacterial activity. The synthetic Ser(P)149 κ-casein-A(138–158) displayed growth-inhibitory activity against S. mutans (MIC, 59 μg/ml [26 μM]). The nonphosphorylated peptide, however, did not inhibit growth at the concentrations tested, indicating that phosphorylation is essential for activity. PMID:11451690

  15. Analysis and Risk Assessment of Seven Toxic Element Residues in Raw Bovine Milk in China.

    PubMed

    Qu, Xue-Yin; Zheng, Nan; Zhou, Xue-Wei; Li, Song-Li; Wang, Jia-Qi; Zhang, Wen-Ju

    2017-08-19

    The object of this study is to analyze the levels of seven toxic elements residues in raw bovine milk in China and assess the potential health risk of those residues. The 178 raw bovine milk samples were collected from eight main milk-producing provinces and from three types of milk stations in China, and were analyzed for arsenic (As), lead (Pb), cadmium (Cd), chromium (Cr), mercury (Hg), aluminum (Al), and nickel (Ni) using inductively coupled plasma-mass spectrometry (ICP-MS). Al, Pb, Hg, Ni, Cr, and As were detected in 47.8, 29.2, 28.1, 23.6, 12.4, and 9.0% of total milk samples, respectively, and Cd were not detected in all samples. The raw bovine milk samples with high levels of toxic elements were found in industrial areas, such as Heilongjiang and Shanxi. Nemerow pollution index analysis showed that the levels were lower in the samples from the processing plants than that from the large-scale farms and small farm cooperatives. The margin of exposure (MOE) values suggest that the levels of As, Pb, Hg, Cr, Al, and Ni in the raw milk samples are not causing a health risk for Chinese consumers, including adults and children. Nevertheless, the risk of Pb for infant and young children was more serious than adult.

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

    PubMed

    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.

  17. Pseudomonas aeruginosa lectin PA-IIL as a powerful probe for human and bovine milk analysis.

    PubMed

    Lesman-Movshovich, E; Gilboa-Garber, N

    2003-07-01

    Milk composition exhibits species-specific differences depending on genetic, evolutionary, and environmental factors. In addition, commercial milk preparations are also changed by industrial manipulations, including severe heat processing. Cow milk, used as human food, provides important nutrients but lacks some essential components that are present in raw human milk. The present study, which was aimed at comparing infant breastfeeding to cow-based formula nourishment, shows major differences between the human and the commercial cow milk glycans detectable by the lectins PA-IL (galactose-binding) and PA-IIL (fucose and mannose-binding) isolated from the cells of human pathogen Pseudomonas aeruginosa. More than 40 human milk samples, several cow milks, and bovine milk-based infant formulas, were examined using these two lectins. For purposes of comparison, the plant lectins Concanavalin A (Con A), which binds mannose, and Ulex europaeus 1st lectin (UEA-I), which binds fucose, were also used. The most prominent difference was revealed using PA-IIL, which displayed a unique high sensitivity to the human milk fucosylated compounds. PA-IL and UEA-I also exhibited preferential sensitivity to the human milk but considerably lower than that of PA-IIL. Con A was inhibited by human and the other milk preparations examined to the same extent. These findings indicate the superb applicability of PA-IIL for rapid and reliable comparative investigation of milk glycans from human and cow, indicating which glycans could be added to infant formulas in order to enrich them, as well as for verification and quality control of otherwise improved bovine milk-based infant formulas.

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

    PubMed

    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.

  19. Phagocytosis of bovine blood and milk polymorphonuclear leukocytes after ozone gas administration in vitro.

    PubMed

    Ducusin, Rio John T; Nishimura, Masakazu; Sarashina, Takao; Uzuka, Yuji; Tanabe, Shigeyuki; Otani, Masayuki

    2003-04-01

    To determine the effects of ozone on the phagocytosis of bovine polymorphonuclear leukocytes (PMNs), ozone gas was administered in vitro on the blood and milk of healthy lactating cows, cows with acute mastitis, and cows with milk fever. In the blood of healthy dairy cattle, although there was no significant effect of ozone gas on the viability of the leukocytes, phagocytosis of PMNs significantly decreased. In contrast, ozone gas administration in vitro significantly increased phagocytosis of PMNs from the blood of cows with acute mastitis and milk fever, and from mastitic milk. These findings showed that ozone administration in vitro has positive and negative effects on bovine PMN phagocytosis, depending on the health status of the animal.

  20. Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil.

    PubMed

    Albuquerque, Pedro Paulo Feitosa de; Santos, André de Souza; Souza Neto, Orestes Luiz de; Kim, Pomy de Cássia Peixoto; Cavalcanti, Erika Fernanda Torres Samico Fernandes; Oliveira, Júnior Mário Baltazar de; Mota, Rinaldo Aparecido; Júnior, José Wilton Pinheiro

    The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples.

  1. Identification and quantification of bovine protein lactosylation sites in different milk products.

    PubMed

    Milkovska-Stamenova, Sanja; Hoffmann, Ralf

    2016-02-16

    The microbiological safety of milk is typically guaranteed by thermal treatments, such as pasteurization and ultra high temperature (UHT) treatment, whereas infant formula (IF) is often produced at even harsher conditions including a drying process. Thermal treatments have raised concerns, as they may denature proteins and initiate protein modifications. Previous studies identified already many lactosylation sites in milk and showed that the lactosylation degree of some proteins correlates to thermal treatment conditions. Here, we studied the glycation degrees of 124 lactosylation sites in 28 bovine milk proteins in raw milk, three brands of pasteurized milk, three brands of UHT milk, and five brands of IF. Whereas, the glycation degree of many lactosylation sites increased from raw milk, to pasteurized milk, UHT milk, and IF, several modification sites showed a different behavior indicating that global measures do not correctly reflect the reactivity of distinct sites. Interestingly, the glycation degrees varied considerably among the brands of UHT milk and IF indicating that specific production processes of a company have to be considered and not only the classification of milk as pasteurized or UHT. Thus, proper adjustments of the technical processes should allow reducing the lactosylation levels in both UHT milk and IF. It is well established that thermal treatment of milk triggers protein modifications, such as lactosylation of lysine residues in several proteins, although the extent of lactosylation has not been quantitatively compared for a broad panel of protein lactosylation sites among different commercial products. The current study extends previous reports by relatively quantifying 124 confirmed lactosylation sites in 28 bovine milk proteins including several low abundant proteins. Whereas, glycation is generally assumed to be an unspecific chemical reaction with the modification degrees depending on the protein and sugar concentrations, we could show

  2. Infection of human keratinocytes by Streptococcus dysgalactiae subspecies dysgalactiae isolated from milk of the bovine udder.

    PubMed

    Roma-Rodrigues, Catarina; Alves-Barroco, Cynthia; Raposo, Luís R; Costa, Mafalda N; Fortunato, Elvira; Baptista, Pedro Viana; Fernandes, Alexandra R; Santos-Sanches, Ilda

    2016-04-01

    Streptococcus dysgalactiae subsp. dysgalactiae (SDSD) are considered exclusive animal pathogens; however, a putative zoonotic upper limb cellulitis, a prosthetic joint infection and an infective endocarditis were described in humans. To unravel if bovine SDSD isolates are able to infect human cells, the adherence and internalization to human primary keratinocytes of two bovine SDSD strains isolated from milk collected from udder were analyzed. Bacterial adhesion assays and confocal microscopy indicate a high adherence and internalization of SDSD isolates to human cells, suggesting for the first time the ability of bovine isolates to infect human cells.

  3. Variation in the bovine FABP4 gene affects milk yield and milk protein content in dairy cows

    PubMed Central

    Zhou, H.; Cheng, L.; Azimu, W.; Hodge, S.; Edwards, G. R.; Hickford, J. G. H.

    2015-01-01

    Fatty acid binding proteins (FABPs) bind long-chain fatty acids and are involved in their intracellular transport. Of the known bovine FABP genes, FABP4 has been mapped to a region on chromosome 14 that contains quantitative trait loci for milk traits. This study investigated the association of FABP4 haplotypes with milk production traits in 719 Holstein-Friesian × Jersey cows. Polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis of a variable region of the gene revealed three haplotypes (A, B and C). Five single nucleotide polymorphisms (SNPs) were identified: two in exon 3 and three in intron 3. A was associated (P = 0.032) with increased milk protein percentage (present: 4.00 ± 0.02%; absent: 3.95 ± 0.02%) and B was associated (P = 0.009) with increased milk yield (present: 23.81 ± 0.23 kg/d; absent: 23.06 ± 0.21 kg/d), but tended to be associated with a decrease in protein percentage and an increase in protein yield. Cows with genotypes AA, AB and AC produced less milk, but with a higher protein percentage than BC cows. This suggest that FABP4 affects milk yield and milk protein content, both economically important traits, and that further study of this gene is warranted. PMID:26067182

  4. The effect of bovine milk on the growth of Bombyx mori.

    PubMed

    Konala, Niharika; Abburi, Praveena; Bovilla, Venugopal Reddy; Mamillapalli, Anitha

    2013-01-01

    Bombyx mori L. (Lepidoptera: Bombycidae) is a well-studied Lepidopteran model system because of its morphology, life cycle, and economic importance. Many scientists have placed importance on enhancing the economic traits of B. mori because it's larvae, silkworms, are vital in the production of silk. In this study, the effect of bovine milk on B. mori growth was tested. Bovine milk contains several components that aid in healthy growth. The treatment was given to fifth instar B. mori larvae because the fifth instar period is when B. mori eats voraciously and shows maximum growth among all its larval stages. The larvae were treated with fresh mulberry, Morus L. (Rosales: Moraceae), leaves and mulberry leaves dipped in milk from the first day of the fifth instar. Treatments were given on alternate days, and the silkworms were weighed every day to determine whether milk had any role in enhancing the weight of the larvae. Cocoon weights were measured, as the weight indicates the approximate amount of silk that can be reeled. The results showed that larvae gained 82.5% more weight by the end of fifth instar larval when fed with mulberry leaves dipped in milk than when fed with fresh mulberry leaves without milk. The larvae fed with milk-treated leaves gained 310% weight from day 1 to day 7 of the fifth instar, while the larvae fed with fresh leaves gained 153% weight in the same timespan. In addition, cocoon weight increased by 8% when milk was added compared to when it was not. These results suggest that B. mori larvae can be fed mulberry leaves treated with bovine milk for better growth rate and increased silk production.

  5. The Effect of Bovine Milk on the Growth of Bombyx mori

    PubMed Central

    Konala, Niharika; Abburi, Praveena; Bovilla, Venugopal Reddy; Mamillapalli, Anitha

    2013-01-01

    Bombyx mori L. (Lepidoptera: Bombycidae) is a well-studied Lepidopteran model system because of its morphology, life cycle, and economic importance. Many scientists have placed importance on enhancing the economic traits of B. mori because it's larvae, silkworms, are vital in the production of silk. In this study, the effect of bovine milk on B. mori growth was tested. Bovine milk contains several components that aid in healthy growth. The treatment was given to fifth instar B. mori larvae because the fifth instar period is when B. mori eats voraciously and shows maximum growth among all its larval stages. The larvae were treated with fresh mulberry, Morus L. (Rosales: Moraceae), leaves and mulberry leaves dipped in milk from the first day of the fifth instar. Treatments were given on alternate days, and the silkworms were weighed every day to determine whether milk had any role in enhancing the weight of the larvae. Cocoon weights were measured, as the weight indicates the approximate amount of silk that can be reeled. The results showed that larvae gained 82.5% more weight by the end of fifth instar larval when fed with mulberry leaves dipped in milk than when fed with fresh mulberry leaves without milk. The larvae fed with milk-treated leaves gained 310% weight from day 1 to day 7 of the fifth instar, while the larvae fed with fresh leaves gained 153% weight in the same timespan. In addition, cocoon weight increased by 8% when milk was added compared to when it was not. These results suggest that B. mori larvae can be fed mulberry leaves treated with bovine milk for better growth rate and increased silk production. PMID:24205942

  6. Detection of Helicobacter pylori in bovine, buffalo, camel, ovine, and caprine milk in Iran.

    PubMed

    Rahimi, Ebrahim; Kheirabadi, Elahe Kazemi

    2012-05-01

    Helicobacter pylori infection in humans is one of the most common infections worldwide. However, the origin and transmission of this bacterium has not been clearly explained. One of the suggested theories is transmission via raw milk from animals to human beings. This study was conducted to determine the prevalence rate of H. pylori in bulk milk samples from dairy bovine, buffalo, camel, ovine, and caprine herds in Iran. In the present study, 447 bulk milk samples from 230 dairy bovine, buffalo, camel, ovine, and caprine herds were collected in four provinces and tested for H. pylori by cultural method and polymerase chain reaction (PCR) for the detection of the ureC (glmM) gene. The animals whose milk samples collected for this study were clinically healthy. Using the cultural method, three of 447 milk samples (0.67%), including two sheep (2.2%) and one buffalo (1.6%) milk samples, were found to be contaminated with H. pylori. H. pylori ureC gene was detected in 56 (12.5%) of milk samples, including 19 cow (14.1%), 11 sheep (12.2%), nine goat (8.7%), two camel (3.6%), and 15 buffalo (23.4%) milk samples. Using PCR method, there were significant differences (p<0.05) in the level of contamination with H. pylori between milk samples collected from different species. The present study is the first report of the isolation of H. pylori from raw sheep and buffalo milk in Iran and the first demonstration of H. pylori DNA in camel and buffalo milk.

  7. Interaction of a green ester-bonded gemini surfactant with xanthine oxidase: Biophysical perspective.

    PubMed

    Akram, Mohd; Bhat, Imtiyaz Ahmad; Kabir-ud-Din

    2015-01-01

    A multi technique approach was utilized to explore the interaction between a novel green gemini surfactant, ethane-1,2-diyl bis(N,N-dimethyl-N-tetradecylammoniumacetoxy) dichloride (14-E2-14), with bovine milk xanthine oxidase (XO). Tensiometric, spectroscopic, microscopic and molecular modeling results demonstrate significant interaction and structural change of native xanthine oxidase upon 14-E2-14 combination. The results obtained in this study may be beneficial for scientists to calibrate conformation of the enzyme by novel biodegradable/green microstructures; consequently, it would likely add new impetus in understanding the treatment modes of various diseases like gout, hyperuricemia, liver and brain necrosis. Moreover, the 14-E2-14-XO interaction assists to unfurl new routes in the designing/selection of green-surfactant-protein mixtures widely used in food processing, cosmetics, and pharmaceutical industries. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Bovine milk exosomes contain microRNA and mRNA and are taken up by human macrophages.

    PubMed

    Izumi, Hirohisa; Tsuda, Muneya; Sato, Yohei; Kosaka, Nobuyoshi; Ochiya, Takahiro; Iwamoto, Hiroshi; Namba, Kazuyoshi; Takeda, Yasuhiro

    2015-05-01

    We reported previously that microRNA (miRNA) are present in whey fractions of human breast milk, bovine milk, and rat milk. Moreover, we also confirmed that so many mRNA species are present in rat milk whey. These RNA were resistant to acidic conditions and to RNase, but were degraded by detergent. Thus, these RNA are likely packaged in membrane vesicles such as exosomes. However, functional extracellular circulating RNA in bodily fluids, such as blood miRNA, are present in various forms. In the current study, we used bovine raw milk and total RNA purified from exosomes (prepared by ultracentrifugation) and ultracentrifuged supernatants, and analyzed them using miRNA and mRNA microarrays to clarify which miRNA and mRNA species are present in exosomes, and which species exist in other forms. Microarray analyses revealed that most mRNA in milk whey were present in exosomes, whereas miRNA in milk whey were present in supernatant as well as exosomes. The RNA in exosomes might exert functional effects because of their stability. Therefore, we also investigated whether bovine milk-derived exosomes could affect human cells using THP-1 cells. Flow cytometry and fluorescent microscopy studies revealed that bovine milk exosomes were incorporated into differentiated THP-1 cells. These results suggest that bovine milk exosomes might have effects in human cells by containing RNA.

  9. Contribution of aldehyde oxidase, xanthine oxidase, and aldehyde dehydrogenase on the oxidation of aromatic aldehydes.

    PubMed

    Panoutsopoulos, Georgios I; Kouretas, Demetrios; Beedham, Christine

    2004-10-01

    Aliphatic aldehydes have a high affinity toward aldehyde dehydrogenase activity but are relatively poor substrates of aldehyde oxidase and xanthine oxidase. In addition, the oxidation of xenobiotic-derived aromatic aldehydes by the latter enzymes has not been studied to any great extent. The present investigation compares the relative contribution of aldehyde dehydrogenase, aldehyde oxidase, and xanthine oxidase activities in the oxidation of substituted benzaldehydes in separate preparations. The incubation of vanillin, isovanillin, and protocatechuic aldehyde with either guinea pig liver aldehyde oxidase, bovine milk xanthine oxidase, or guinea pig liver aldehyde dehydrogenase demonstrated that the three aldehyde oxidizing enzymes had a complementary substrate specificity. Incubations were also performed with specific inhibitors of each enzyme (isovanillin for aldehyde oxidase, allopurinol for xanthine oxidase, and disulfiram for aldehyde dehydrogenase) to determine the relative contribution of each enzyme in the oxidation of these aldehydes. Under these conditions, vanillin was rapidly oxidized by aldehyde oxidase, isovanillin was predominantly metabolized by aldehyde dehydrogenase activity, and protocatechuic aldehyde was slowly oxidized, possibly by all three enzymes. Thus, aldehyde oxidase activity may be a significant factor in the oxidation of aromatic aldehydes generated from amines and alkyl benzenes during drug metabolism. In addition, this enzyme may also have a role in the catabolism of biogenic amines such as dopamine and noradrenaline where 3-methoxyphenylacetic acids are major metabolites.

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

    PubMed

    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.

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

    PubMed

    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.

  12. Seasonal variation of polar lipid content in bovine milk.

    PubMed

    Liu, Zhiqian; Logan, Amy; Cocks, Benjamin G; Rochfort, Simone

    2017-12-15

    Seasonal change of milk composition could offer opportunities for dairy manufacturers. A systematic survey on seasonal variation of six classes of polar lipids was conducted with 19 Holstein cows over the entire milking season using liquid chromatography-mass spectrometry technique. This study revealed that most polar lipid classes were positively correlated with the total fat content, but negatively correlated with fat globule size. All polar lipid classes displayed a large cow-to-cow variation as well as seasonal variation. All of the six classes showed a gradual increase over the milking season with the highest concentration observed in May (autumn). However, the proportion of different polar lipid classes remained constant during the entire milking season. This finding suggests that the production of polar lipids is highly regulated in the mammary gland. The implication of such a seasonal variation of polar lipids in the nutritional and technological aspects of milk is discussed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Peptidomic analysis reveals proteolytic activity of kefir microorganisms on bovine milk proteins

    PubMed Central

    Dallas, David C.; Citerne, Florine; Tian, Tian; Silva, Vitor L. M.; Kalanetra, Karen M.; Frese, Steven A.; Robinson, Randall C.; Mills, David A.; Barile, Daniela

    2015-01-01

    Scope The microorganisms that make up kefir grains are well known for lactose fermentation, but the extent to which they hydrolyze and consume milk proteins remains poorly understood. Peptidomics technologies were used to examine the proteolytic activity of kefir grains on bovine milk proteins. Methods and results Gel electrophoresis revealed substantial digestion of milk proteins by kefir grains, with mass spectrometric analysis showing the release of 609 protein fragments and alteration of the abundance of >1,500 peptides that derived from 27 milk proteins. Kefir contained 25 peptides identified from the literature as having biological activity, including those with antihypertensive, antimicrobial, immunomodulatory, opioid and anti-oxidative functions. 16S rRNA and shotgun metagenomic sequencing identified the principle taxa in the culture as Lactobacillus species. Conclusion The model kefir sample contained thousands of protein fragments released in part by kefir microorganisms and in part by native milk proteases. PMID:26616950

  14. Peptidomic analysis reveals proteolytic activity of kefir microorganisms on bovine milk proteins.

    PubMed

    Dallas, David C; Citerne, Florine; Tian, Tian; Silva, Vitor L M; Kalanetra, Karen M; Frese, Steven A; Robinson, Randall C; Mills, David A; Barile, Daniela

    2016-04-15

    The microorganisms that make up kefir grains are well known for lactose fermentation, but the extent to which they hydrolyze and consume milk proteins remains poorly understood. Peptidomics technologies were used to examine the proteolytic activity of kefir grains on bovine milk proteins. Gel electrophoresis revealed substantial digestion of milk proteins by kefir grains, with mass spectrometric analysis showing the release of 609 protein fragments and alteration of the abundance of >1500 peptides that derived from 27 milk proteins. Kefir contained 25 peptides identified from the literature as having biological activity, including those with antihypertensive, antimicrobial, immunomodulatory, opioid and anti-oxidative functions. 16S rRNA and shotgun metagenomic sequencing identified the principle taxa in the culture as Lactobacillus species. The model kefir sample contained thousands of protein fragments released in part by kefir microorganisms and in part by native milk proteases. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

    SciTech Connect

    Ang, C.Y.W.; Luo, Wenhong

    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.

  16. Using milk leukocyte differentials for diagnosis of subclinical bovine mastitis.

    PubMed

    Gonçalves, Juliano Leonel; Lyman, Roberta L; Hockett, Mitchell; Rodriguez, Rudy; Dos Santos, Marcos Veiga; Anderson, Kevin L

    2017-08-01

    This research study aimed to evaluate the use of the milk leukocyte differential (MLD) to: (a) identify quarter milks that are culture-positive; and (b) characterize the milk leukocyte responses to specific groups of pathogens causing subclinical mastitis. The MLD measures the absolute number and relative percentage of inflammatory cells in milk samples. Using the MLD in two dairy herds (170 and 172 lactating cows, respectively), we studied all lactating cows with a most recent monthly Dairy Herd Improvement Association somatic cell count (SCC) >200 × 103 cells/ml. Quarter milk samples from 78 cows meeting study criteria were analysed by MLD and aseptically collected milk samples were subjected to microbiological culture (MC). Based upon automated instrument evaluation of the number and percentage of inflammatory cells in milk, samples were designated as either MLD-positive or - negative for subclinicial mastitis. Positive MC were obtained from 102/156 (65·4%) of MLD-positive milk samples, and 28/135 (20·7%) of MLD-negative milk samples were MC-positive. When MC was considered the gold standard for mastitis diagnosis, the calculated diagnostic Se of the MLD was 65·4% (IC95% = 57·4 to 72·8%) and the Sp was 79·3% (IC95% = 71·4 to 85·7%). Quarter milks positive on MC had higher absolute numbers of neutrophils, lymphocytes and macrophages, with higher neutrophils% and lymphocytes% but lower macrophages%. The Log10 (N/L) ratios were the most useful ratio to differentiate specific subclinical mastitis quarters from healthy quarters. Use of the MLD on cows with monthly composite SCC > 200 × 103 cells/ml for screening at quarter level identified quarters more likely to be culture-positive. In conclusion, the MLD can provide an analysis of mammary quarter status more detailed than provided by SCC alone; however, the MLD response to subclinical mastitis was not found useful to specifically identify the causative pathogen.

  17. Effect of infection with bovine leukemia virus on milk production in Michigan dairy cows.

    PubMed

    Norby, B; Bartlett, P C; Byrem, T M; Erskine, R J

    2016-03-01

    The objective of this study was to determine the association between individual cow-level milk production and bovine leukemia virus (BLV) infection as measured by milk BLV-ELISA. Dairy Herd Improvement technicians collected milk samples from 10 cows from each of first, second, third, and 4+ parity cows in 105 Holstein herds with ≥ 120 milking cows. Milk samples were tested for the presence of anti-BLV antibodies by ELISA. Additional data regarding the cows and the herds were collected by farm survey and Dairy Herd Improvement records. A set of mixed-effect models using all cows and only 2+ parity cows were used to investigate the association between BLV ELISA-corrected optical density and 305-d mature equivalents of individual cows. The BLV milk positivity was associated with decreased 305-d mature-equivalent yields, especially among the older cows. Additionally, increasing milk ELISA-corrected optical density was associated with increasing loss of milk production at the cow level. In summary, our results provide evidence that BLV infection is associated with decreased milk production in Michigan dairy cows. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

    PubMed

    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.

  19. Estimation of heritability and genetic correlations for the major fatty acids in bovine milk.

    PubMed

    Soyeurt, H; Gillon, A; Vanderick, S; Mayeres, P; Bertozzi, C; Gengler, N

    2007-09-01

    The current cattle selection program for dairy cattle in the Walloon region of Belgium does not consider the relative content of the different fatty acids (FA) in milk. However, interest by the local dairy industry in differentiated milk products is increasing. Therefore, farmers may be interested in selecting their animals based on the fat composition. The aim of this study was to evaluate the feasibility of genetic selection to improve the nutritional quality of bovine milk fat. The heritabilities and correlations among milk yield, fat, protein, and major FA contents in milk were estimated. Heritabilities for FA in milk and fat ranged from 5 to 38%. The genetic correlations estimated among FA reflected the common origin of several groups of FA. Given these results, an index including FA contents with the similar metabolic process of production in the mammary gland could be used, for example, to increase the monounsaturated and conjugated fatty acids in milk. Moreover, the genetic correlations between the percentage of fat and the content of C14:0, C12:0, C16:0, and C18:0 in fat were -0.06, 0.55, 0.60, and 0.84, respectively. This result demonstrates that an increase in fat content is not directly correlated with undesirable changes in FA profile in milk for human health. Based on the obtained genetic parameters, a future selection program to improve the FA composition of milk fat could be initiated.

  20. Dose response of lactating dairy ewes during suckling and milking to bovine somatotropin.

    PubMed

    Requena, R; Balasch, S; Peris, C; Rodríguez, M; Fernández, N

    2010-09-01

    The objectives were to determine the effect of administering recombinant bovine ST (bST) every 14 d on milk yield and milk composition in dairy ewes and to assess the possible effects of treatment on milk yield loss that occurs at lamb weaning. Seventy-two lactating dairy ewes were separated into 4 groups at lambing, and each group received no bST (control) or 40, 80, or 120 mg of bST every 14 d until wk 20 of lactation. During the first 5 wk of lactation, ewes suckled their lambs during the night and were milked once daily (period 1). After weaning, from 6 to 22 wk of lactation, ewes were exclusively milked twice daily (period 2). Actual milk yield, potential milk yield, and milk component percentages were recorded weekly throughout lactation. In period 1, bST-treated groups increased yields of potential milk (P = 0.04) and the corresponding 6% fat-corrected milk (FCM; P = 0.04) but not actual milk yield (P = 0.42) compared with the untreated group. In period 2, treated groups increased potential (P < 0.01) and actual (P < 0.01) milk yields as well as their corresponding 6% FCM (potential, P < 0.01; actual, P < 0.01), in comparison with the untreated group. The group treated with 120 mg of bST every 14 d yielded 39% more actual milk and 44% more 6% FCM than the control group for period 2. At weaning, between the last week of period 1 and the first week of period 2, treated groups showed similar absolute (P = 0.15) and relative (P = 0.33) values for the potential milk losses compared with the control group. Treatments increased (P = 0.04) actual milk fat percentages during period 1, but did not affect the other milk components or somatic cell count. In summary, bST increased potential milk yield throughout lactation and actual milk yield only after weaning in dairy ewes. In dairy ewes, bST was not useful for reducing the milk yield loss that occurred at weaning.

  1. An association between milk and slime increases biofilm production by bovine Staphylococcus aureus.

    PubMed

    Fabres-Klein, Mary Hellen; Caizer Santos, Mário Junior; Contelli Klein, Raphael; Nunes de Souza, Guilherme; de Oliveira Barros Ribon, Andrea

    2015-01-16

    Staphylococcus aureus is associated with chronic mastitis in cattle, and disease manifestation is usually refractory to antibiotic therapy. Biofilm production is a key element of S. aureus pathogenesis and may contribute to the treatment failure that is consistently reported by veterinarians. Minas Gerais State is the largest milk-producing state in Brazil, and the characterization of bacterial isolates is an important aspect of disease control for dairy farmers. Here, we investigated the potential of S. aureus isolated from bovine mastitis to produce slime and biofilm in a skim-milk medium and classified the isolates according to their agr type. Slime was detected using the Congo Red agar (CRA) test in 35.18% (19/54) of the strains; however, 87.04% (47/54) of the strains were considered biofilm-positive based on crystal violet staining. Compared to TSB supplemented with 0.25% glucose, skim milk significantly increased the production of biofilm, but this effect was only observed in slime-producing strains. The bacteria belonged to agr groups I (12/54), II (34/54), III (6/54), and IV (2/54), and bacteria in agr group III were found to be stronger biofilm producers than those in groups I and II. Again, milk had a significant influence only on slime-positive agr I and II isolates, revealing an association between milk and slime. The present study demonstrated that skim-milk medium and slime production are two factors that together influence biofilm formation by bovine strains of S. aureus. A predominance of bacteria belonging to agr group II was observed, and bacteria from agr group III showed the highest proportion of biofilm producers. The majority of bacteria characterized in this study formed biofilm in milk, which suggests that biofilm formation has an important role in the virulence of S. aureus isolated from bovine intramammary infections.

  2. The function of the milk-clotting enzymes bovine and camel chymosin studied by a fluorescence resonance energy transfer assay.

    PubMed

    Jensen, Jesper Langholm; Jacobsen, Jonas; Moss, Marcia L; Rasmussen, Fred; Qvist, Karsten Bruun; Larsen, Sine; van den Brink, Johannes M

    2015-05-01

    Enzymatic coagulation of bovine milk can be divided in 2 steps: an enzymatic step, in which the Phe105-Met106 bond of the milk protein bovine κ-casein is cleaved, and an aggregation step. The aspartic peptidases bovine and camel chymosin (EC 3.4.23.4) are typically used to catalyze the enzymatic step. The most commonly used method to study chymosin activity is the relative milk-clotting activity test that measures the end point of the enzymatic and aggregation step. This method showed that camel chymosin has a 2-fold higher milk-clotting activity toward bovine milk than bovine chymosin. To enable a study of the enzymatic step independent of the aggregation step, a fluorescence resonance energy transfer assay has been developed using a peptide substrate derived from the 98-108 sequence of bovine κ-casein. This assay and Michaelis-Menten kinetics were employed to determine the enzymatic activity of camel and bovine chymosin under milk clotting-like conditions (pH 6.65, ionic strength 80 mM). The results obtained show that the catalytic efficiency of camel chymosin is 3-fold higher than bovine chymosin. The substrate affinity and catalytic activity of bovine and camel chymosin increase at lower pH (6.00 and 5.50). The glycosylation of bovine and camel chymosin did not affect binding of the fluorescence resonance energy transfer substrate, but doubly glycosylated camel chymosin seems to have slightly higher catalytic efficiency. In the characterization of the enzymes, the developed assay is easier and faster to use than the traditionally used relative milk-clotting activity test method.

  3. Proteolytic activity of Enterococcus faecalis VB63F for reduction of allergenicity of bovine milk proteins.

    PubMed

    Biscola, V; Tulini, F L; Choiset, Y; Rabesona, H; Ivanova, I; Chobert, J-M; Todorov, S D; Haertlé, T; Franco, B D G M

    2016-07-01

    With the aim of screening proteolytic strains of lactic acid bacteria to evaluate their potential for the reduction of allergenicity of the major bovine milk proteins, we isolated a new proteolytic strain of Enterococcus faecalis (Ent. faecalis VB63F) from raw bovine milk. The proteases produced by this strain had strong activity against caseins (αS1-, αS2-, and β-casein), in both skim milk and sodium caseinate. However, only partial hydrolysis of whey proteins was observed. Proteolysis of Na-caseinate and whey proteins, observed after sodium dodecyl sulfate-PAGE, was confirmed by analysis of peptide profiles by reversed-phase HPLC. Inhibition of proteolysis with EDTA indicated that the proteases produced by Ent. faecalis VB63F belonged to the group of metalloproteases. The optimal conditions for their activity were 42°C and pH 6.5. The majority of assessed virulence genes were absent in Ent. faecalis VB63F. The obtained results suggest that Ent. faecalis VB63F could be efficient in reducing the immunoreactivity of bovine milk proteins.

  4. Annotation and structural elucidation of bovine milk oligosaccharides and determination of novel fucosylated structures

    PubMed Central

    Aldredge, Danielle L; Geronimo, Maria R; Hua, Serenus; Nwosu, Charles C; Lebrilla, Carlito B; Barile, Daniela

    2013-01-01

    Bovine milk oligosaccharides (BMOs) are recognized by the dairy and food industries, as well as by infant formula manufacturers, as novel, high-potential bioactive food ingredients. Recent studies revealed that bovine milk contains complex oligosaccharides structurally related to those previously thought to be present in only human milk. These BMOs are microbiotic modulators involved in important biological activities, including preventing pathogen binding to the intestinal epithelium and serving as nutrients for a selected class of beneficial bacteria. Only a small number of BMO structures are fully elucidated. To better understand the potential of BMOs as a class of biotherapeutics, their detailed structure analysis is needed. This study initiated the development of a structure library of BMOs and a comprehensive evaluation of structure-related specificity. The bovine milk glycome was profiled by high-performance mass spectrometry and advanced separation techniques to obtain a comprehensive catalog of BMOs, including several novel, lower abundant neutral and fucosylated oligosaccharides that are often overlooked during analysis. Structures were identified using isomer-specific tandem mass spectroscopy and targeted exoglycosidase digestions to produce a BMO library detailing retention time, accurate mass and structure to allow their rapid identification in future studies. PMID:23436288

  5. Annotation and structural elucidation of bovine milk oligosaccharides and determination of novel fucosylated structures.

    PubMed

    Aldredge, Danielle L; Geronimo, Maria R; Hua, Serenus; Nwosu, Charles C; Lebrilla, Carlito B; Barile, Daniela

    2013-06-01

    Bovine milk oligosaccharides (BMOs) are recognized by the dairy and food industries, as well as by infant formula manufacturers, as novel, high-potential bioactive food ingredients. Recent studies revealed that bovine milk contains complex oligosaccharides structurally related to those previously thought to be present in only human milk. These BMOs are microbiotic modulators involved in important biological activities, including preventing pathogen binding to the intestinal epithelium and serving as nutrients for a selected class of beneficial bacteria. Only a small number of BMO structures are fully elucidated. To better understand the potential of BMOs as a class of biotherapeutics, their detailed structure analysis is needed. This study initiated the development of a structure library of BMOs and a comprehensive evaluation of structure-related specificity. The bovine milk glycome was profiled by high-performance mass spectrometry and advanced separation techniques to obtain a comprehensive catalog of BMOs, including several novel, lower abundant neutral and fucosylated oligosaccharides that are often overlooked during analysis. Structures were identified using isomer-specific tandem mass spectroscopy and targeted exoglycosidase digestions to produce a BMO library detailing retention time, accurate mass and structure to allow their rapid identification in future studies.

  6. Identification of lipid synthesis and secretion proteins in bovine milk.

    PubMed

    Lu, Jing; van Hooijdonk, Toon; Boeren, Sjef; Vervoort, Jacques; Hettinga, Kasper

    2014-02-01

    Lactation physiology is a process that is only partly understood. Proteomics techniques have shown to be useful to help advance the knowledge on lactation physiology in human and rodent species but have not been used as major tools for dairy cows, except for mastitis. In this paper, advanced non-targeted proteomics techniques (Filter aided sample preparation and NanoLC-Orbitrap-MS/MS) were applied to study the milk fat globule membrane and milk serum fraction, resulting in the identification of 246 proteins. Of these, 23 transporters and enzymes were related to lipid synthesis and secretion in mammary gland and their functions are discussed in detail. The identification of these intracellular transporters and enzymes in milk provides a possibility of using milk itself to study lipid synthesis and secretion pathways. This full-scale scan of milk proteins by using non-targeted proteomic analysis helps to reveal the important proteins involved in lipid synthesis and secretion for further examination in targeted studies.

  7. Reduction in heat-induced gastrointestinal hyperpermeability in rats by bovine colostrum and goat milk powders.

    PubMed

    Prosser, C; Stelwagen, K; Cummins, R; Guerin, P; Gill, N; Milne, C

    2004-02-01

    Male Sprague-Dawley rats were assigned to one of three dietary groups [standard diet (Cont; n = 8), standard diet plus bovine colostrum powder (BColost 1.7 g/kg; n = 8), or goat milk powder (GMilk 1.7 g/kg; n = 8)] to determine the ability of these supplements to reduce gastrointestinal hyperpermeability induced by heat. Raising core body temperature of rats to 41.5 degrees C increased transfer of (51)Cr-EDTA from gut into blood 34-fold relative to the ambient temperature value (P < 0.05) in the Cont group of rats, indicative of increased gastrointestinal permeability. Significantly less (P < 0.01) (51)Cr-EDTA was transferred into the blood of rats in either the BColost (27% of Cont) or GMilk group (10% of Cont) after heating, showing that prior supplementation with either bovine colostrum or goat milk powder significantly reduced the impact of heat stress on gastrointestinal permeability. The changes in the BColost group were not significantly different than those of the GMilk group. The potential mechanism of the protective effect of bovine colostrum and goat milk powders may involve modulation of tight junction permeability, because both powders were able to maintain transepithelial resistance in Madin Darby canine kidney cells challenged with EGTA compared with cells maintained in media only. The results show that bovine colostrum powder can partially alleviate the effects of hyperthermia on gastrointestinal permeability in the intact animal. Moreover, goat milk powder was equally as effective as bovine colostrum powder, and both may be of benefit in other situations where gastrointestinal barrier function is compromised.

  8. Comparative study on heat stability and functionality of camel and bovine milk whey proteins.

    PubMed

    Laleye, L C; Jobe, B; Wasesa, A A H

    2008-12-01

    Heat stability, emulsifying, and foaming properties of camel whey have been investigated and compared with that of bovine whey. Camel whey is similar to bovine whey in composition, but is deficient in beta-lactoglubulin (beta-LG), a major component of bovine whey. Whether the deficiency in beta-LG will affect stability and functional properties is not yet known. Substantial information on the functional properties of bovine milk whey proteins is available; however, there is little research done on functional properties of camel whey proteins. Therefore, the objective of this study was to investigate the heat stability, emulsifying, and foaming characteristics of camel whey proteins. Calorimetric studies showed no significant difference in heat stability between bovine and camel whey proteins in liquid form. Upon drying, thermograms indicated that the 2 proteins are different in composition and thermal stability. The difference is represented in the absence of beta-LG and the occurrence of protein denaturation peak at a lesser temperature in camel whey. The first marginal thermal transition in bovine whey appeared at 81 degrees C, followed by 2 other transitions at 146 and 198 degrees C. For camel whey, the transitions appeared at 139, 180, and 207 degrees C respectively. The first marginal denaturation peak in bovine whey is due to beta-LG, which is essentially absent in camel whey, while the second peak is due to the mixture of alpha-lactalbumin, serum albumin, and possibly part of the partially stabilized beta-LG structure during the denaturation process. Because camel whey is deficient in beta-LG, the denaturation peak at 139 must be due to the mixture of alpha-lactalbumin and camel serum albumin. In both proteins, the highest thermal transition is due to sugars such as lactose. The solubility study has shown that camel whey is more sensitive to pH than bovine milk whey and that heat stability is lowest near the isoelectric point of the proteins at pH 4.5. The

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

    PubMed

    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.

  10. Elevated milk soluble CD14 in bovine mammary glands challenged with Escherichia coli lipopolysaccharide.

    PubMed

    Lee, J W; Paape, M J; Elsasser, T H; Zhao, X

    2003-07-01

    The purpose of this study was to determine whether soluble CD14 (sCD14) in milk was affected by stage of lactation, milk somatic cell count (SCC), presence of bacteria, or lipopolysaccharide (LPS)-induced inflammation. Milk samples from 100 lactating cows (396 functional quarters) were assayed for sCD14 in milk to determine effects of stage of lactation, SCC, and intramammary infection. The concentration of sCD14 was highest in transitional milk (0 to 4 d postpartum) and in milk with high SCC (> 750,000 cells/ml). Most of the infected quarters (> 80%) were infected by coagulase-negative staphylococci and yeast. No difference was found between noninfected and infected quarters. One quarter of six healthy lactating cows was challenged with 100 microg LPS in order to study the kinetics of sCD14 during an LPS-induced inflammation. Milk samples were collected at various intervals until 72 h after injection. Rectal temperature, milk tumor necrosis factor-alpha, and interleukin-8 increased immediately after challenge. The increase in sCD14 paralleled the increase in SCC, peaked at 12 h, and started to decline after 24 h. Serum leakage, as characterized by the level of bovine serum albumin in milk, peaked at 4 h and then gradually decreased. All parameters remained at basal levels in control quarters throughout the study. In vitro experiments indicated that neutrophils released sCD14 in response to LPS in a dose-dependent manner. The results indicate that the concentration of sCD14 was significantly increased in milk after LPS challenge. The increase was not likely due to serum leakage. Instead, infiltrated neutrophils might be the main source of increased sCD14 in milk during inflammation.

  11. Measurement of bovine somatotropin (bST) and insulin-like growth factor-1 (IGF-1) in bovine milk using an electrochemiluminescent assay.

    PubMed

    McGrath, Michael F; Bogosian, Gregg; Fabellar, Anabella C; Staub, Robin L; Vicini, John L; Widger, Leslie A

    2008-08-27

    Bovine somatotropin (bST) and insulin-like growth factor-1 (IGF-1) are peptide hormones that are involved in the regulation of milk production in dairy cows. Because these hormones are present at extremely low concentration in fresh and processed bovine milk, a highly sensitive and specific electrochemiluminescent immunoassay (ECLIA) has been developed to better estimate the concentration of these hormones in milk. The assay employs an imager, a capture antibody bound to a carbon electrode, and a detection antibody coupled to a ruthenium label. In the presence of tripropylamine and an electric pulse, ruthenium generates light proportional to the amount of antigen bound, and the light is captured as signal by a charge-coupled device (CCD) camera. Using bovine milk as the starting matrix, 99.69% of bST and 104.79% of IGF-1 were recoverable. The limit of detection (LOD) was <5 pg/mL for bST and <1 pg/mL for IGF-1. The limit of quantification (LOQ) was <14 pg/mL for bST in milk and <2 pg/mL of IGF-1. The assay is highly specific and shows <0.2% cross-reactivity with other peptide hormones found in bovine milk such as insulin and IGF-2. These data indicate this new, ECLIA is highly sensitive and specific for estimating the concentration of bST or IGF-1 in milk.

  12. Antibiotic susceptibilities and prevalence of Methicillin resistant Staphylococcus aureus (MRSA) isolated from bovine milk in Pakistan.

    PubMed

    Aqib, Amjad Islam; Ijaz, Muhammad; Anjum, Aftab Ahmad; Malik, Muhammad Abdul Rauf; Mehmood, Khalid; Farooqi, Shahid Hussain; Hussain, Kashif

    2017-08-07

    The study was designed to investigate bovine milk for prevalence of an emerging zoonotic pathogen Methicillin resistant Staphylococcus aureus (MRSA), and in-vitro therapeutic response of various antibiotics against MRSA. Nine hundred (900) milk samples were collected (half from cattle and half from buffalo) from private and public farms located in various tehsils of district Faisalabad, using the convenient sampling method. Milk samples were put to biochemical identification of Staphylococcus aureus and later oxacilline disk sensitivity testing for confirmation of MRSA. The MRSA isolates were confirmed by PCR targeting mecA gene in Staphylococcus aureus. The study found 34% prevalence of MRSA in overall bovine milk from district Faisalabad with 30% and 38% prevalence in cattle and buffalo, respectively. Tehsil Samundari presented comparatively higher MRSA prevalence followed by tehsil Jaranwala and tehsil Faisalabad. However, there was non-significant difference of MRSA prevalence between cattle and buffalo, and among different tehsils. All assumed risk factors except specie were significantly associated with mastitis spread. The in-vitro drug trial against MRSA from buffalo milk presented 100% efficacy of Ciprofloxcin, Moxifloxacin, Linezolid, and Trimethoprim plus Sulphamethoxazole combination, followed by Gentamicin and Levofloxacin presenting 90%, and Amikacin becoming 80% efficacious against MRSA from buffalo milk. The MRSA isolates of cattle milk presented similar pattern with some variations of higher susceptibility against Oxytetracycline, and Fusidic acid. The conclusion of the study states uniform prevalence of MRSA in cattle and buffalo milk in study area having assumed risk factors positively associated with disease spread, while Ciprofloxcin, Moxifloxacin, Linezolid, and Trimethoprim plus Sulphamethoxazole drugs showed the highest efficacy to combat this pathogen. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. An amperometric biosensor for xanthine determination prepared from xanthine oxidase immobilized in polypyrrole film.

    PubMed

    Arslan, Fatma; Yaşar, Ahmet; Kiliç, Esma

    2006-01-01

    In order to prepare a biosensor for the determination of xanthine, electropolymerization of pyrrole on Pt surface was carried out with an electrochemical cell containing pyrrole, ferrocene (as a electron mediator) and tetrabutylamonium tetrafluoroborat in acetonitrile by cyclic voltammetry between 0.0 and 0.9V (vs SCE) at a scan rate of 50mV/s upon Pt electrode. Xanthine oxidase was immobilized by a glutaraldehyde/bovine serum albumin (BSA) crosslinking procedure on to polypyrrole film after the electropolymerization processes. The response of the biosensor against xanthine was measured after 3-4 min following the application of a constant potential of + 0.7 V (vs SCE). The resulting biosensor exhibits excellent electrocatalysis for the xanthine. The amperometric determination is based on the electrochemical detection of H202, which is generated in enzymatic reaction of xanthine. The effect of various experimental conditions was examined for the determination of the analytical performance. The sensor responds to xanthine with a detection limit of 1.0 x 10(-6)M. The response current increases linearly with xanthine concentration up to 4.0 x 10(-4) M. The sensor remains relatively stable for 45 days.

  14. Association between the bovine milk metabolome and rennet-induced coagulation properties of milk.

    PubMed

    Sundekilde, Ulrik K; Gustavsson, Frida; Poulsen, Nina A; Glantz, Maria; Paulsson, Marie; Larsen, Lotte B; Bertram, Hanne C

    2014-10-01

    The milk metabolomes of 407 individual Swedish Red dairy cows were analyzed by nuclear magnetic resonance spectroscopy as part of the Danish-Swedish Milk Genomics Initiative. By relating these metabolite profiles to total milk protein concentration and rheological measurements of rennet-induced milk coagulation together using multivariate data analysis techniques, we were able to identify several different associations of the milk metabolome to technological properties of milk. Several novel correlations of milk metabolites to protein content and rennet-induced coagulation properties were demonstrated. Metabolites associated with the prediction of total protein content included choline, N-acetyl hexosamines, creatinine, glycerophosphocholine, glutamate, glucose 1-phosphate, galactose 1-phosphate, and orotate. In addition, levels of lactate, acetate, glutamate, creatinine, choline, carnitine, galactose 1-phosphate, and glycerophosphocholine were significantly different when comparing noncoagulating and well-coagulating milks. These findings suggest that the mentioned metabolites are associated with milk protein content and rennet-induced coagulation properties and may act as quality markers for cheese milk. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

    PubMed

    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.

  16. Microfluidic sedimentation cytometer for milk quality and bovine mastitis monitoring.

    PubMed

    Garcia-Cordero, Jose L; Barrett, Louise M; O'Kennedy, Richard; Ricco, Antonio J

    2010-12-01

    We report a rapid, low-cost, portable microfluidic sedimentation cytometer (SeCy) for assessing the somatic cell count and fat content of milk in 15 min using a "sample-in, answer-out" approach. The system consists of 12 independent microfluidic devices, essentially flattened funnel structures, fabricated on the footprint of a single plastic compact disc (CD). Each funnel structure holds 150 μL of milk, has an inlet for milk filling and an outlet for air to escape, and ends in a narrow, closed-end microfluidic channel that facilitates packing of the cells into a column whose length is proportional to cell count. The closed-end channel provides accurate cell counts over the range 50,000->3,000,000 cells per mL. The assay separates cells and fat globules based on their densities (by differential sedimentation), concentrating white cells in the closed-end channel near the outer rim of the CD for estimation of total "cell pellet" volume, while fat globules move toward the center of disc rotation, forming a fat "band" in the funnel. After adding milk to two or more microfluidic devices, the CD is loaded onto a custom-built reader unit that spins the disc for 15 min. Two low-cost microscopes in the reader image the centrifuged cell pellet and the fat band, providing a sufficiently accurate cell count to diagnose mastitis and measuring fat content as an indication of health and nutritional status.

  17. Growth is similar in premature infants fed human milk fortified with a human milk or bovine milk-based fortifier

    USDA-ARS?s Scientific Manuscript database

    Preterm infants require fortification of human milk (HM) to provide adequate nutrients for growth. A human milk fortifier (HMF) made from donor human milk has recently been introduced. However, limited data are available comparing the growth outcomes of this product with current fortifiers. We evalu...

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

    PubMed

    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.

  19. Development of magnetic nanoparticle based calorimetric assay for the detection of bovine mastitis in cow milk.

    PubMed

    Chinnappan, Raja; Al Attas, Sana; Kaman, Wendy E; Bikker, Floris J; Zourob, Mohammed

    2017-04-15

    Mastitis in dairy cattle is an inflammatory reaction of the udder tissue. Mastitis increases plasmin levels, leading to an increased proteolysis of milk proteins such as casein, resulting in a significant decrease in milk quality and related dairy products. Due to its key-role in mastitis, we used plasmin proteolytic activity as a biomarker for the detection of mastitis in bovine mastitic milk. Inspired by earlier studies on protease activity using mastitic milk samples, we developed a simple colorimetric assay to distinguish mastitic milk from milk derived from healthy animals. The plasmin substrate coupled to magnetic nanoparticles form a black self-assembled monolayer on a gold sensor surface. In the presence of increased levels of plasmin, the substrate is cleaved and the peptide fragment attached to the magnetic beads, will be attracted by the magnet which is present under the sensor strips revealing the golden surface. We found the area of the golden color surface proportional to plasmin activity. The sensitivity of this method was determined to be 1 ng/ml of plasmin in vitro. Next, we tested the biosensor using mastitis positive milk of which infection is confirmed by bacterial cultures. This newly developed colorimetric biosensor has high potential in applications for the diagnosis of mastitis with potential spin offs to health, food and environmental sectors. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Effect of different levels of supplied cobalt on the fatty acid composition of bovine milk.

    PubMed

    Karlengen, Inger J; Taugbøl, Ole; Salbu, Brit; Aastveit, Are H; Harstad, Odd M

    2013-03-14

    In previous studies, administration of high amounts of Co decreased the proportion of MUFA in bovine milk. The present study was conducted to examine the amount of Co needed to obtain this effect. High-yielding dairy cows (n 4), equipped with ruminal cannulas, were used in a 4 × 4 Latin square design study. The basal diet consisted of concentrate mixture (9 kg/d) without added Co and grass silage (ad libitum). The following four levels of Co were administrated as cobalt acetate dissolved in distilled water: no Co (treatment 1, T1); 4·0 mg Co/d (T2); 380 mg Co/d (T3); 5300 mg Co/d (T4). Each period lasted for 18 d, including 11 d of treatment. During the treatment periods, the solutions were continuously infused into the rumen. Milk yield and milk concentration of fat, fatty acids (FA), protein, lactose, Co, Zn, Fe and Cu were determined. Blood plasma was analysed with respect to FA, Co, Zn, Fe and Cu. Feed intake and total tract digestibility of feed components were also determined. There was a linear effect of increasing the level of Co on milk FA composition. The effects of Co on FA composition in blood were insignificant compared with the effects on milk. In milk fat, the concentration of cis-9-18 : 1 was reduced by as much as 38 % on T4 compared with T1. Feed intake and milk yield were negatively affected by increasing the Co level.

  1. Relationship between somatic cell count, polymorphonuclear leucocyte count and quality parameters in bovine bulk tank milk.

    PubMed

    Wickström, Erik; Persson-Waller, Karin; Lindmark-Månsson, Helena; Ostensson, Karin; Sternesjö, Ase

    2009-05-01

    The somatic cell count (SCC) in bovine bulk tank milk is presently used as an indicator of raw milk quality, reflecting the udder health status of the herd. During mastitis, SCC increases, mostly owing to an influx of polymorphonuclear leucocytes (PMN) from blood into milk, with a concomitant change in milk composition. Bulk tank milk samples were categorized according to their SCC, as well as polymorphonuclear leucocyte count (PMNC), to study relationships between SCC, PMNC and various raw milk quality traits, i.e. contents of total protein, whey protein, casein, fat and lactose, casein number, proteolysis and rheological properties. The proportion of PMN, obtained by direct microscopy, was significantly higher in samples with high SCC compared with low SCC samples. SCC and PMNC were strongly correlated, yielding a correlation coefficient of 0.85. High SCC samples had lower lactose and casein contents, lower casein number and more proteolysis than low SCC samples. Samples with high PMNC had a lower casein number than low PMNC samples. Samples with high and low SCC or PMNC did not differ in respect to rheological properties. Our results do not indicate that PMNC is a better biomarker than SCC for raw bulk tank milk quality, as previously proposed.

  2. Occurrence and characterization of methicillin-resistant staphylococci from bovine mastitis milk samples in Finland

    PubMed Central

    2013-01-01

    Background Methicillin-resistant staphylococci (MRS) are increasingly being isolated in bovine mastitis. The aim of our study was to evaluate the occurrence of MRS in Finnish mastitis milk samples and characterize the MRS isolates using molecular methods. Results Methicillin-resistant S. aureus (MRSA) was a rare finding in bovine mastitis in Finland. Only two out of 135 (1.5%) S. aureus isolates were positive for mec genes. One of these carried mecA and was of spa type t172, SCCmec type IV and ST375, and the other harboured mecC, being spa type t3256, and ST130. MRSA ST375 is common among human MRSA isolates in Finland, but this is the first report in the country of bovine mecC MRSA. In coagulase-negative staphylococci (CoNS) originating from bovine mastitis, methicillin resistance was more common. In the two CoNS collections studied, 5.2% (17/324) and 1.8% (2/110) of the isolates were mecA positive. Eighteen of these were methicillin-resistant S. epidermidis (MRSE), which were divided into 6 separate PFGE clusters. One pulsotype was detected in different parts of the country, indicating clonal spread. Most MRSE (13/18) were of SCCmec type IV, one was of type V and four were non-typeable. Comparison with a human staphylococcal database indicated that bovine MRSE strains were not closely related to human MRSE isolates. Conclusions The occurrence of MRS, especially MRSA, in bovine mastitis in Finland was low. Most methicillin-resistant bovine CoNS are MRSE, and we found evidence of a bovine MRSE strain that may spread clonally. This is the first report of a Finnish bovine isolate of MRSAmecC ST130. The study provides a baseline for further MRS monitoring. PMID:23985065

  3. Occurrence and characterization of methicillin-resistant staphylococci from bovine mastitis milk samples in Finland.

    PubMed

    Gindonis, Veera; Taponen, Suvi; Myllyniemi, Anna-Liisa; Pyörälä, Satu; Nykäsenoja, Suvi; Salmenlinna, Saara; Lindholm, Laura; Rantala, Merja

    2013-08-28

    Methicillin-resistant staphylococci (MRS) are increasingly being isolated in bovine mastitis. The aim of our study was to evaluate the occurrence of MRS in Finnish mastitis milk samples and characterize the MRS isolates using molecular methods. Methicillin-resistant S. aureus (MRSA) was a rare finding in bovine mastitis in Finland. Only two out of 135 (1.5%) S. aureus isolates were positive for mec genes. One of these carried mecA and was of spa type t172, SCCmec type IV and ST375, and the other harboured mecC, being spa type t3256, and ST130. MRSA ST375 is common among human MRSA isolates in Finland, but this is the first report in the country of bovine mecC MRSA. In coagulase-negative staphylococci (CoNS) originating from bovine mastitis, methicillin resistance was more common. In the two CoNS collections studied, 5.2% (17/324) and 1.8% (2/110) of the isolates were mecA positive. Eighteen of these were methicillin-resistant S. epidermidis (MRSE), which were divided into 6 separate PFGE clusters. One pulsotype was detected in different parts of the country, indicating clonal spread. Most MRSE (13/18) were of SCCmec type IV, one was of type V and four were non-typeable. Comparison with a human staphylococcal database indicated that bovine MRSE strains were not closely related to human MRSE isolates. The occurrence of MRS, especially MRSA, in bovine mastitis in Finland was low. Most methicillin-resistant bovine CoNS are MRSE, and we found evidence of a bovine MRSE strain that may spread clonally. This is the first report of a Finnish bovine isolate of MRSAmecC ST130. The study provides a baseline for further MRS monitoring.

  4. Prevalence of bovine milk pathogens in Azorean pastures: mobile versus fixed milking machines

    PubMed Central

    Azevedo, C.; Pacheco, D.; Soares, L.; Moitoso, M.; Maldonado, J.; Guix, R.

    2016-01-01

    The aims of the present study were (1) to evaluate the influence of using mobile (n=47) or fixed (n=45) milking machines in Azorean herds on the apparent prevalence of several milk pathogens in bulk tank milk (BTM) and (2) to determine whether separated subclinical mastitic cows can serve, in real time, as predictors of milk pathogen prevalence for the remaining animals at the herd level. The use of a mobile or fixed milking machine influenced (P≤0.05) the prevalence of Staphylococcus aureus (72.3 per cent; n=34 v 51.1 per cent; n=23, respectively) and Klebsiella species (46.8 per cent; n=22 v 26.7 per cent; n=12, respectively). S aureus (95 per cent CI OR 1.1 to 6.0) and Klebsiella species (95 per cent CI OR 1.0 to 5.8) were 2.5 times more likely to increase in the BTM of herds using mobile milking machines. The prevalence of coagulase-negative staphylococci (100 per cent; n=92), Escherichia coli (75.0 per cent), Corynebacterium bovis (57.6 per cent), Enterococcus species (55.4 per cent), Streptococcus dysgalactiae (51.1 per cent), Streptococcus uberis (41.3 per cent), Actinomyces pyogenes or Peptostreptococcus indolicus (41.3 per cent) and Streptococcus agalactiae (32.6 per cent) in BTM remained similar among the herds. κ coefficients were always <0.70, indicating intra-herd disagreement of the prevalence of milk pathogens between BTM and separated milking cows. Milking hygiene should be improved in pastures, focusing specifically on herds that use a mobile milking machine. The segregated cows at milking time are not good predictors of milk pathogens in BTM. PMID:27843558

  5. [Bovine milk progesterone concentration as a method of early diagnosis of pregnancy (author's transl)].

    PubMed

    Guissi, G; Laborde, N P; Rosner, J M

    1976-01-01

    With the purpose of obtaining an early indicator of pregnancy in bovines, excretion levels of milk progesterone were determined by a simple radioimmunological method. In the pregnant group, progesterone concentration was significantly (p less than 0,001) greater than in non-pregnant ones. In some cases (20-25%) there was no difference in values, resulting a method efficiency of 75-80%. Milk must be processed a few hours after collection since a decrease in progesterone values is observed within a few days. Easy obtention of this biological matherial, without special technical requirements as well as the simple dosification method make this test a practical index for establishing early diagnosis of pregnancy in bovines.

  6. Streptomycin and dihydrostreptomycin residues in bovine milk from the Brazilian retail market.

    PubMed

    de Oliveira, Renata C; Paschoal, Jonas A R; Reyes, Felix G R

    2010-01-01

    Pasteurised bovine milk from retail markets in the State of São Paulo, Brazil, was analysed for the presence of streptomycin (STP) and dihydrostreptomycin (DHSTP) residues. An ELISA kit was used for screening and a LC-APCI-MS/MS QToF method for confirmatory analysis. Both methods were intra-laboratory validated and found suitable for screening and confirmatory testing, respectively, for STP and DHSTP residues in pasteurised bovine milk at concentration levels below the maximum residue limit (MRL) established for these substances (200 µg kg(-1) expressed as the sum of the concentrations of STP and DHSTP). No residues of STP and DHSTP at detectable levels were found in the analysed samples (n = 299).

  7. Evaluation of milk cathelicidin for detection of bovine mastitis.

    PubMed

    Addis, M F; Tedde, V; Puggioni, G M G; Pisanu, S; Casula, A; Locatelli, C; Rota, N; Bronzo, V; Moroni, P; Uzzau, S

    2016-10-01

    Mastitis due to intramammary infection is one of the most economically relevant diseases in dairy cows, causing reductions in milk quality and quantity. Currently, mastitis monitoring is based on somatic cell count (SCC) and bacteriologic culture (BC) of milk. Nevertheless, inflammation-specific protein markers might provide more sensitive and reliable assays, enabling immunoassay-based screening strategies. Cathelicidin is an inflammatory protein released in milk that has recently demonstrated fair reliability and diagnostic potential for ewe mastitis. To assess its performance in cows, 531 quarter milk samples from 2 herds were tested using cathelicidin ELISA, SCC, and BC. We found that 29.0% of samples were positive for cathelicidin, 18.8% had SCC >200,000 cells/mL, and 13.7% were BC-positive. Cathelicidin showed a strong positive correlation with SCC as demonstrated by receiver operating characteristics curve analysis and by the clustering of cathelicidin-negative and cathelicidin-positive samples in association with low and high SCC values, respectively. For evaluating the diagnostic performance of a novel test, BC cannot be considered a reliable gold standard for true disease status because of its known limitations. Therefore, we assessed the sensitivity (Se) and specificity (Sp) of the milk cathelicidin ELISA using a latent class analysis approach together with BC and SCC by considering different diagnostic thresholds to identify the preferred Se/Sp combination. We modeled conditional dependence of cathelicidin and SCC to account for their close association. The cathelicidin ELISA showed higher Se than SCC and BC for almost all threshold combinations. In fact, at the best-performing threshold combination, the Se of cathelicidin was 80.6%, 6.2 percentage points higher than that of SCC >200,000 cells/mL (74.4%) and similar to that of SCC >100,000 cells/mL (80.2%). Most importantly, this Se was obtained with a loss in Sp of only 1.4 percentage points compared

  8. Exposure assessment of dioxins and dioxin-like PCBs in pasteurised bovine milk using probabilistic modelling.

    PubMed

    Adekunte, Adefunke O; Tiwari, Brijesh K; O'Donnell, Colm P

    2010-09-01

    Quantitative exposure assessment is a useful technique to investigate the risk from contaminants in the food chain. The objective of this study was to develop a probabilistic exposure assessment model for dioxins (PCDD/Fs) and dioxin-like PCBs (DL-PCBs) in pasteurised bovine milk. Mean dioxins and DL-PCBs (non-ortho and mono-ortho PCBs) concentrations (pg WHO-TEQ g(-1)) in bovine milk were estimated as 0.06 ± 0.07 pg WHO-TEQ g(-1) for dioxins and 0.08 ± 0.07 pg WHO-TEQ g(-1) for DL-PCBs using Monte Carlo simulation. The simulated model estimated mean exposure for dioxins was 0.19 ± 0.29 pg WHO-TEQ kg(-1)bw d(-1) and 0.14 ± 0.22 pg WHO-TEQ kg(-1) bw d(-1) and for DL-PCBs was 0.25 ± 0.30 pg WHO-TEQ kg(-1) bw d(-1) and 0.19 ± 0.22 pg WHO-TEQ kg(-1) bw d(-1) for men and women, respectively. This study showed that the mean dioxins and DL-PCBs exposure from consumption of pasteurised bovine milk is below the provisional maximum tolerable monthly intake of 70 pg TEQ kg(-1) bw month(-1) (equivalent of 2.3 pg TEQ kg(-1) bw d(-1)) recommended by the Joint FAO/WHO Expert Committee on Food Additives and Contaminants (JECFA). Results from this study also showed that the estimated dioxins and DL-PCBs concentration in pasteurised bovine milk is comparable to those reported in previous studies. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Antioxidant properties and xanthine oxidase inhibitory effects of Tamus communis L. root extracts.

    PubMed

    Boumerfeg, Sabah; Baghiani, Abderrahmane; Messaoudi, Dalila; Khennouf, Seddik; Arrar, Lekhmici

    2009-02-01

    This study was conducted to search for xanthine oxidase (XO) inhibitors from the root extracts of Tamus communis L. traditionally used in folk medicine in Algeria. Root extracts with different solvents were screened for purified milk xanthine oxidase inhibition. The root extracts (methanol, chloroform and ethyl acetate) and proteins, obtained in distilled water, inhibited bovine, sheep and human milk XO from three species in a concentration-dependent manner, with an additional superoxide scavenging capacity, which reached its highest level with ethyl acetate extract (IC(50) = 0.15, 0.04 and 0.09 g/L) for bovine XO, sheep XO and human XO, respectively. The antioxidant potential was confirmed with the non-enzymatic method, total radical-trapping antioxidant parameter (TRAP) assay, which showed that the Tamus communis L. extracts have a potential antioxidant activity in the same order obtained by using the reduction of cytochrome c, an enzymatic method, in which the antioxidant activity followed a decreasing order: ethyl acetate extract > chloroform extract > protein.

  10. Variation in fat globule size in bovine milk and its prediction using mid-infrared spectroscopy.

    PubMed

    Fleming, A; Schenkel, F S; Chen, J; Malchiodi, F; Ali, R A; Mallard, B; Sargolzaei, M; Corredig, M; Miglior, F

    2017-03-01

    The objectives of this study were to investigate the sources of variation in milk fat globule (MFG) size in bovine milk and its prediction using mid-infrared (MIR) spectroscopy. Mean MFG size was measured in 2,076 milk samples from 399 Ayrshire, Brown Swiss, Holstein, and Jersey cows, and expressed as volume moment mean (D[4,3]) and surface moment mean (D[3,2]). The mid-infrared spectra of the samples and milk performance data were also recorded during routine milk recording and testing. The effects of breed, herd nested within breed, days in milk, season, milking period, age at calving, parity, and individual animal on the variation observed in MFG size were investigated. Breed, herd nested within breed, days in milk, season, and milking period significantly affected mean MFG size. Milk fat globule size was the largest at the beginning of lactation and subsequently decreased. Milk samples with the smallest MFG on average came from Holstein cows, and those with the largest were from Jersey and Brown Swiss cows. Partial least squares regression was used to predict MFG size from MIR spectra of samples with a calibration data set containing 2,034 and 2,032 samples for D[4,3] and D[3,2], respectively. Coefficients of determination of cross validation for D[4,3] and D[3,2] prediction models were 0.51 and 0.54, respectively. The associated ratio of performance deviation values were 1.43 and 1.48 for D[4,3] and D[3,2], respectively. With these models, individual mean MFG size could not be accurately predicted, but results may be sufficient to screen samples for having either small or large MFG on average. Significant but low correlations of D[4,3] and D[3,2] with milk fat yield were estimated (0.16 and 0.21, respectively). Significant and moderate Pearson correlation coefficients for fat percent with D[4,3] and D[3,2] were assessed (0.34 and 0.36, respectively). This correlation was greater between milk fat percentage and predicted MFG size than with measured MFG size

  11. Innate immune response to a bovine mastitis pathogen profiled in milk and blood monocytes using a systems biology approach

    USDA-ARS?s Scientific Manuscript database

    Bovine mastitis is an inflammatory condition of the mammary gland which leads to reduced milk yield and increased milk somatic cell counts (SCC) resulting in an estimated annual cost to the dairy industry worldwide of ~ 2 billion euros. Mastitis has a complex etiology, with pathogenic, host and envi...

  12. Molecular identification and thermoresistance to boiling of Nocardia farcinica and Nocardia cyriacigeorgica from bovine bulk tank milk

    PubMed Central

    Condas, L.A.Z.; Ribeiro, M.G.; Gonoi, T.; Matsuzawa, T.; Yazawa, K.; Motta, R.G.; Franco, M.M.J.; Listoni, F.J.P.

    2012-01-01

    Two strains of Nocardia spp. were isolated from bovine milk of two individual bulk tank. Molecular identification classified the strains as Nocardia farcinica and Nocardia cyriacigeorgica. The thermorresistance to boiling of the isolates was carried out and was observed bacterial growth after boiling. Our findings indicate the potential risk of pathogen transmission to humans through contaminated milk with Nocardia spp. PMID:24031926

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

    PubMed

    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.

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

    PubMed

    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

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

    PubMed

    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.

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

    PubMed Central

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

    2014-01-01

    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. We also observed a corresponding decrease in the saturation content of the raw cream phospholipids, and a loss of phosphatidylethanolamine after spray-drying to produce buttermilk powder. PMID:20828196

  17. Human milk components cross-reacting with antibodies against bovine beta-lactoglobulin.

    PubMed

    Neuteboom, B; Giuffrida, M G; Cantisani, A; Napolitano, L; Alessandri, A; Fabris, C; Bertino, E; Conti, A

    1992-01-01

    The human whey components cross-reacting with antibodies raised against bovine and/or equine beta-lactoglobulin were screened systematically. The milk of six women on a normal diet was collected within 72 h of confinement and whey components were fractionated by high-speed size exclusion chromatography and reversed-phase techniques. The fractions which were immunoreactive in double diffusion experiments with antisera anti-bovine and/or equine beta-lactoglobulin were subsequently purified by native PAGE and then electroblotted on Pro-blott membrane (Western blotting). Pro-blot membranes were stained in parallel with Coomassie and by immunostaining using antibodies against bovine and/or equine beta-lactoglobulin as first antibody solution. The immunoreactive bands were cut out from the membrane and N-terminally sequenced; all the immunoreactive components were clearly identified as human beta-casein or its (mainly tryptic) fragments. The strong antigenic similarity between human beta-casein and beta-lactoglobulin (bovine and equine) might be of immunological importance; it could mean that breast-fed neonates risk being sensitized to beta-lactoglobulin irrespective of the presence of cow's milk in the mother's diet.

  18. Effect of long-term bovine somatotropin (sometribove) treatment on nitrogen (protein) distribution in Jersey milk.

    PubMed

    Kindstedt, P S; Pell, A N; Rippe, J K; Tsang, D S; Hartnell, G F

    1991-01-01

    Twenty-six Jersey cows were assigned randomly to one of two treatments. Twelve cows received biweekly subcutaneous injection of 500 mg of sometribove, USAN (recombinant methionyl bovine somatotropin), beginning 60 +/- 3 d postpartum and continuing throughout one lactation. Fourteen control animals received injections of placebo carrier. Milk samples were taken biweekly on weeks alternate to injection when differences in milk components were expected to be greatest compared with controls. The milk samples were analyzed for total nitrogen, noncasein nitrogen, and non-protein nitrogen. The average SCC for control and treatment groups was 44,000 +/- 47,000 and 56,000 +/- 65,000. Milk from sometribove-treated cows was significantly lower in total protein (3.92, 4.12%), true protein (3.74, 3.95%), and casein (3.11, 3.34%) than that from control cows on d 8 of the 14 d injection cycle. Casein as a percentage of true protein was lower (83.38, 84.52%), and non-protein nitrogen as a percentage of total nitrogen was higher (4.61, 4.26%) in milk from treated cows. The theoretical yield of Cheddar cheese was ca. .07% less for milk from treated cows than from control cows due to ca. 1% less casein as a percentage of true protein in the former. The differences in nitrogen distribution represent the response during the middle of the injection cycle when milk output was the highest and milk protein the lowest rather than the average response for the injection cycle. The results of the study indicate minimal impact on the cheese manufacturer because in practice milk is commingled from many dairies.(ABSTRACT TRUNCATED AT 250 WORDS)

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

    PubMed

    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 <0.05) in 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.

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

    PubMed Central

    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 <0.05) in 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. PMID:27100870

  1. Clinical mastitis in cows treated with sometribove (recombinant bovine somatotropin) and its relationship to milk yield.

    PubMed

    White, T C; Madsen, K S; Hintz, R L; Sorbet, R H; Collier, R J; Hard, D L; Hartnell, G F; Samuels, W A; de Kerchove, G; Adriaens, F

    1994-08-01

    Effect of sometribove (methionyl bovine somatotropin) on mastitis in 15 full lactation trials (914 cows) in Europe and the US and 70 short-term studies (2697 cows) in eight countries was investigated. In full lactation studies, sometribove (500 mg/2 wk) was given for 252 d, commencing 60 d postpartum. Although herds varied considerably, incidence of clinical mastitis within a herd was similar for cows receiving control and sometribove treatments. Relative risk analyses indicated no treatment effect, and percentage of mastitis during treatment was similar for control and sometribove groups. A positive linear relationship existed between peak milk yield and mastitis incidence (percentage of cows contracting mastitis or cases per 100 cow days); sometribove treatment did not alter this relationship. Increases in mastitis related to milk yield increase from sometribove or related to genetic selection were similar. When expressed per unit of milk, mastitis incidence declined slightly as milk yield increased; this relationship was not altered by sometribove. No effect on clinical mastitis was observed in 70 commercial herds utilizing sometribove for 84 d. However, effects were significant for stage of lactation and milk yield. Overall, studies represented a wide range of research and commercial situations demonstrating that sometribove had no effect on incidence of clinical mastitis during the lactation of treatment. Furthermore, sometribove did not alter typical relationships between milk yield or herd factors and incidence of clinical mastitis.

  2. Determination of imidocarb residues in bovine and ovine liver and milk by immunobiosensor.

    PubMed

    Traynor, I M; Thompson, C S; Armstrong, L; Fodey, T; Danaher, M; Jordan, K; Kennedy, D G; Crooks, S R H

    2013-01-01

    Imidocarb (IMD) is a veterinary drug that has been used for more than 30 years to treat and prevent parasitic diseases. Pharmacokinetic studies have shown that substantial levels of IMD residues are retained in the edible tissues and milk of cattle and sheep for up to 6 months after administration. This has led to concern regarding the potential adverse effects posed through human consumption of edible tissue or milk from treated animals if the recommended withdrawal periods for the drug are not properly implemented. While MRLs have been established by the European Union, it is important that analytical methods are available to monitor food samples for potentially violative levels of IMD residues. A qualitative biosensor-based immunoassay was developed to allow the detection of IMD at less than the European Union MRLs of 50 μg kg(-1) for milk and 2 mg kg(-1) for bovine and ovine liver. Validation of the developed methods provided a detection capability of <25 μg kg(-1) in milk and <0.75 mg kg(-1) in liver. A comparison study was undertaken, with IMD incurred milk and ovine liver samples analysed by the newly developed procedures and results compared with those obtained by LC-MS/MS. The newly developed screening method was applied to both incurred milk and liver samples. This faster, cheaper and reliable screening method has potential use in sample analysis to ensure compliance with legislative requirements.

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

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  5. The effect of a cannula milk sampling technique on the microbiological diagnosis of bovine mastitis.

    PubMed

    Friman, M; Hiitiö, H; Niemi, M; Holopainen, J; Pyörälä, S; Simojoki, H

    2017-08-01

    Two methods of collecting milk samples from mastitic bovine mammary quarters were compared. Samples were taken in a consistent order in which standard aseptic technique sampling was done first, followed by insertion of a sterile cannula through the teat canal and collection of a second sample. Microbiological results of those two sampling techniques were compared. Milk samples were analysed using multiplex real-time polymerase chain reaction (PCR). The cannula technique produced a reduced number of microbial species or groups of species per sample compared with conventional sampling. Staphylococcus spp. were the most common species identified and were detected more often during conventional sampling than with cannula sampling. Staphylococcus spp. identified in milk samples could also have originated from the teat canal without being present in the milk. The number of samples positive for Trueperella pyogenes or yeasts in the conventional samples was twice as high as in the cannula samples, indicating that the presence of Trueperella pyogenes and yeast species should not necessarily be interpreted as being the causative agents of bovine intra-mammary infections (IMI). Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    PubMed

    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.

  7. Milk and fat yields decline in bovine leukemia virus-infected Holstein cattle with persistent lymphocytosis.

    PubMed

    Da, Y; Shanks, R D; Stewart, J A; Lewin, H A

    1993-07-15

    Effects of bovine leukemia virus (BLV) infection on milk and fat yields were studied by using data collected from Holstein cows over a 6-year period. Milk and fat yields in BLV-infected cows with persistent lymphocytosis (PL) declined significantly relative to their BLV-infected non-PL herdmates. Declines were most pronounced in cows older than 6 years. The estimated loss to the dairy industry due to PL is more than $42 million annually. A major histocompatibility complex class I (BoLA-A) allele that has been previously associated with resistance to PL was associated with longevity and realization of milk production potentials, indicating that genetic resistance to PL will have an economic benefit in herds where BLV is endemic.

  8. Characterization of the bovine milk proteome in early-lactation Holstein and Jersey breeds of dairy cows.

    PubMed

    Tacoma, Rinske; Fields, Julia; Ebenstein, David B; Lam, Ying-Wai; Greenwood, Sabrina L

    2016-01-01

    Milk is a highly nutritious natural product that provides not only a rich source of amino acids to the consumer but also hundreds of bioactive peptides and proteins known to elicit health-benefitting activities. We investigated the milk protein profile produced by Holstein and Jersey dairy cows maintained under the same diet, management and environmental conditions using proteomic approaches that optimize protein extraction and characterization of the low abundance proteins within the skim milk fraction of bovine milk. In total, 935 low abundance proteins were identified. Gene ontology classified all proteins identified into various cellular localization and function categories. A total of 43 low abundance proteins were differentially expressed between the two dairy breeds. Bioactive proteins involved in host-defense, including lactotransferrin (P=0.0026) and complement C2 protein (P=0.0001), were differentially expressed by the two breeds, whereas others such as osteopontin (P=0.1788) and lactoperoxidase (P=0.2973) were not. This work is the first to outline the protein profile produced by two important breeds of dairy cattle maintained under the same diet, environment and management conditions in order to observe likely true breed differences. This research now allows us to better understand and contrast further research examining the bovine proteome that includes these different breeds. Within the last decade, the amount of research characterizing the bovine milk proteome has increased due to growing interest in the bioactive proteins that are present in milk. Proteomic analysis of low abundance whey proteins has mainly focused on human breast milk; however, previous research has highlighted the presence of bioactive proteins in bovine milk. Recent publications outlining the cross-reactivity of bovine bioactive proteins on human biological function highlight the need for further investigation into the bovine milk proteome. The rationale behind this study is to

  9. Assessment of isoflavonoid concentrations in Australian bovine milk samples.

    PubMed

    King, R A; Mano, M M; Head, R J

    1998-08-01

    There is considerable interest in the possibility that diet-derived isoflavonoids may help in protection against a number of chronic diseases common in Western society. Based on animal studies, however, concerns have been raised that consumption of isoflavonoids by infants and young children may be undesirable. Clover contains isoflavonoids and therefore may represent, via milk, a source of isoflavonoids in the human diet. In this study the concentrations of daidzein (7, 4'-dihydroxyisoflavone), genistein (5, 7, 4'-trihydroxyisoflavone) and equol (7-hydroxy-(4'-hydroxyphenyl)chroman) were measured using HPLC in cows' milk samples obtained from 76 farms in three Australian states. In addition, concentrations were measured in samples collected from one South Australian factory both before and after pasteurization. Concentrations in all samples were found to be extremely low. The mean daidzein concentration was < 5 ng/ml. Mean genistein concentrations ranged from just detectable (approximately 2 ng/ml) in Victorian samples collected during summer to 20-30 ng/ml in samples from all states collected during spring when isoflavonoid-containing clover is most dominant in pasture. Mean equol concentrations ranged from 45 +/- 10 ng/ml in Victorian farm samples collected during summer to 293 +/- 52 ng/ml in Western Australian samples collected in spring. The mean concentrations of genistein and equol in post-pasteurization samples collected in spring were approximately double those for samples collected in autumn. Pasteurization had no effect on isoflavonoid concentrations. We conclude that the concentrations of isoflavonoids in Australian cows' milk are low and are therefore unlikely to have any pronounced biological effects in human consumers.

  10. A comparative study of bovine blood and milk neutrophil functions with luminol-dependent chemiluminescence.

    PubMed

    Mehrzad, J; Dosogne, H; Vangroenweghe, F; Burvenich, C

    2001-01-01

    In this study, a technique was developed for the chemiluminescence (CL) measurement of bovine milk polymorphonuclear leukocytes (PMN). In the first study, the effects of cell number and the concentration of phorbol-12-myristate-13-acetate (PMA), luminol, latex bead particles, dimethyl sulphoxide (DMSO) and gelatin on the luminol-dependent cellular CL (LDCL) response were assessed with healthy cows in different stages of lactation. In the second study, the LDCL and in vitro bactericidal activity of blood and milk PMN towards Staphylococcus aureus was investigated. In general, the CL activity of blood PMN was consistently higher than that of milk PMN. We found that (a) the optimal cell density in blood and milk cells for maximal LDCL response ranged from 1.5 x 10(6) to 5 x 10(6) cells/mL; (b) the optimal concentrations of PMA, latex beads and luminol for maximal LDCL response were 100-200 ng/ml, 500 particles/PMN and 0.1 mmol/L, respectively. Concentrations of DMSO of 0.5-1% (v/v) did not significantly affect the maximal CL response of PMN. Gelatin concentrations of 0.1 -0.5 mg/ml had no effect on the LDCL of PMN. In addition, the LDCL of PMN was significantly correlated with bactericidal activity towards S. aureus (r = 0.78, p < 0.001 for blood PMN and r = 0.66, p < 0.01 for milk PMN). Under the optimal experimental conditions for measurement of CL produced by bovine blood and milk PMN defined in this study, LDCL assay is an accurate and reproducible technique for the rapid quantification of PMN bactericidal activity in physiological and pathological conditions of high-yielding dairy cows. Copyright 2001 John Wiley & Sons, Ltd.

  11. Proteinase and phospholipase activities and development at different temperatures of yeasts isolated from bovine milk.

    PubMed

    Melville, Priscilla A; Benites, Nilson R; Ruz-Peres, Monica; Yokoya, Eugenio

    2011-11-01

    The presence of yeasts in milk may cause physical and chemical changes limiting the durability and compromising the quality of the product. Moreover, milk and dairy products contaminated by yeasts may be a potential means of transmission of these microorganisms to man and animals causing several kinds of infections. This study aimed to determine whether different species of yeasts isolated from bovine raw milk had the ability to develop at 37°C and/or under refrigeration temperature. Proteinase and phospholipase activities resulting from these yeasts were also monitored at different temperatures. Five genera of yeasts (Aureobasidium sp., Candida spp., Geotrichum spp., Trichosporon spp. and Rhodotorula spp.) isolated from bovine raw milk samples were evaluated. All strains showed one or a combination of characteristics: growth at 37°C (99·09% of the strains), psychrotrophic behaviour (50·9%), proteinase production (16·81% of the strains at 37°C and 4·09% under refrigeration) and phospholipase production (36·36% of the isolates at 37°C and 10·9% under refrigeration), and all these factors may compromise the quality of the product. Proteinase production was similar for strains incubated at 37°C (16·81% of the isolates) and room temperature (17·27%) but there was less amount of phospholipase-producing strains at room temperature (15·45% of the isolates were positive) when compared with incubation at 37°C (36·36%). Enzymes production at 37°C by yeasts isolated from milk confirmed their pathogenic potential. The refrigeration temperature was found to be most efficient to inhibit enzymes production and consequently ensure better quality of milk. The viability of yeasts and the activity of their enzymes at different temperatures are worrying because this can compromise the quality of dairy products at all stages of production and/or storage, and represent a risk to the consumer.

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

    PubMed

    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.

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

    PubMed

    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.

  14. Protein carbonylation sites in bovine raw milk and processed milk products.

    PubMed

    Milkovska-Stamenova, Sanja; Mnatsakanyan, Ruzanna; Hoffmann, Ralf

    2017-08-15

    During thermal treatment of milk, proteins are oxidized, which may reduce the nutritional value of milk, abolish protein functions supporting human health, especially important for newborns, and yield potentially harmful products. The side chains of several amino acids can be oxidized to reactive carbonyls, which are often used to monitor oxidative stress in organisms. Here we mapped protein carbonylation sites in raw milk and different brands of pasteurized, ultra high temperature (UHT) treated milk, and infant formulas (IFs) after digesting the precipitated proteins with trypsin. Reactive carbonyls were derivatized with O-(biotinylcarbazoylmethyl)hydroxylamine to enrich the modified peptides by avidin-biotin affinity chromatography and analyze them by nanoRP-UPLC-ESI-MS. Overall, 53 unique carbonylated peptides (37 carbonylation sites, 15 proteins) were identified. Most carbonyls were derived from dicarbonyls (mainly glyoxal). The number of carbonylation sites increased with the harsher processing from raw milk (4) to pasteurized (16) and UHT milk (16) and to IF (24). Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    PubMed

    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

  16. Dephosphorylation of bovine casein by milk alkaline phosphatase.

    PubMed

    Lorient, D; Linden, G

    1976-02-01

    The pH of optimum activity of alkaline phosphatase from cow's milk depended on the substrate, being 10-1 for rho-nitrophenylphosphate, 8-6 for phosphoserine, 8-0 for phosvitin and 6-8 for casein. Individual casein components were dephosphorylated more rapidly than mixtures of alphas- and beta-caseins or of alphas-, beta-and kappa-caseins and micellar casein. Mixtures of 2 components involving kappa-casein were more readily dephosphorylated than alphas- and beta-casein mixtures. At pH 6-8, lactose, whey proteins and phosphate ions had an inhibitory effect. beta-Lactoglobulin had an inhibitory effect only when the pH of the reaction was lower than the optimum pH value of the enzyme. Mg2+ and Zn2+ were not inhibitory. The optimum conditions for dephosphorylation of casein are described.

  17. Transdermal delivery of bovine milk vesicles in patients with multiple sclerosis: A novel strategy to induce MOG-specific tolerance.

    PubMed

    Mokarizadeh, Aram; Hassanzadeh, Kambiz; Abdi, Mohammad; Soraya, Hamid; Faryabi, Mohammad Reza; Mohammadi, Ebrahim; Ahmadi, Abbas

    2015-08-01

    Recently, butyrophilin (BTN) - a protein which shares cross-reactive epitopes with myelin oligodendrocyte glycoprotein (MOG) - has been found in milk. A high amount of milk BTN has been reported in the outer membrane of vesicular structures known as exosome and milk fat globule membrane (MFGM). These vesicles can act as Trojan horses, passing their BTN content through epidermis or other biologic barriers of the body. By altering the dose schedule and route of administration, the BTN-bearing vesicles (exosomes and MFGMs) may acquire enough potential to be used in MOG-specific immunotherapy program. Regarding above evidence and considering immunological characteristics of skin-associated lymphoid tissue (SALT), transdermal delivery of bovine milk vesicles, whether through topical administration of bovine milk or by using epicutaneous administration techniques, could be considered as an intriguing approach to induce MOG-specific tolerance in patients with multiple sclerosis.

  18. Insulin in bovine colostrum and milk: evolution throughout lactation and binding to caseins.

    PubMed

    Aranda, P; Sanchez, L; Perez, M D; Ena, J M; Calvo, M

    1991-12-01

    The changes in insulin concentration in bovine milk in the first period of lactation and its association with other milk proteins were studied. Highest concentration was found in the first milking (327 ng/ml). This concentration fell within the first 24 h postpartum to about 50% of its initial value. By d 3, the level was about 25%, and, on d 7, a stable concentration was reached at approximately 46 ng/ml (about 14% of its initial value). This concentration is about 100 times higher than that in serum, which suggests a specific mechanism of transfer from blood to milk. Colostral whey obtained by ultrafiltration or ultracentrifugation contains much less insulin than colostrum. When colostrum or milk was incubated with [125I]insulin and whey and casein fractions were separated by precipitation, it was observed that most insulin remained with the casein. However, when colostrum was incubated with [125I]insulin and subjected to gel filtration, most of the radioactivity corresponded to free insulin, indicating that insulin is associated with the precipitated casein but not with the casein micelles in solution.

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

    PubMed

    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 < 0.05), which suggests the transfer of 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.

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

    PubMed

    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

  1. Lifetime effects of infection with bovine leukemia virus on longevity and milk production of dairy cows.

    PubMed

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

    2016-10-01

    Enzootic bovine leukosis (EBL) is an economically important disease of dairy cattle caused by bovine leukemia virus (BLV). The economic impacts of the infection have been debated in the literature. The present study was conducted to determine the lifetime effects of BLV infection on longevity and milk production of dairy cows in Canada. The data were aggregated from a combination of two data sets: 1) BLV serum-ELISA test results from Canada-wide surveys of production limiting diseases, which took place between 1998 and 2003 in 8 provinces, and 2) longitudinal production data for all cows in the former study, extracted from the Canadian dairy herd improvement database. All participant cows had been culled or died by the onset of this study. A historical cohort study was designed, including cows which tested positive to BLV-antibodies in their first lactation (positive cohort, n=1858) and cows which tested negative in their second or later lactations (negative cohort, n=2194). To assess the impacts of infection with BLV on longevity (the number of lifetime lactations), a discrete-time survival analysis was carried out. The effect of BLV on the lifetime milk production (the sum of all life 305-day milk production) was evaluated using a multilevel linear regression model. Overall, 4052 cows from 348 herds met the eligibility criteria and were enrolled in the study. In the longevity model, the interaction term between time (lactation number) and BLV-status was highly significant. Cows which were positive to BLV had consistently greater probabilities of being culled (or dying) than the test-negative cows. In the milk production model, the interaction term between BLV-status and longevity of the cows was highly significant; indicating that lifetime BLV effects on the total milk production was dependent on the lactation in which the study cows were culled/died. Infected cows with 2 and 3 lactations showed significantly lower life milk productions [-2554kg (-3609 to -1500

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

    PubMed

    Janjanam, Jagadeesh; Jamwal, Manu; Singh, Surender; Kumar, Saravanan; Panigrahi, Aswini K; Hariprasad, Gururao; Jena, Manoj K; Anand, Vijay; Kumar, Sudarshan; Kaushik, Jai K; Dang, Ajay K; Mukesh, Manishi; Mishra, Bishnu P; Srinivasan, Alagiri; Reddy, Vanga S; Mohanty, Ashok K

    2013-11-01

    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.

  3. Simultaneous determination of clavulanic acid and tazobactam in bovine milk by HPLC.

    PubMed

    Guo, Peipei; Chen, Yuan; Yue, Chonghui; Yu, Guoping

    2017-04-01

    A rapid and specific high-performance liquid chromatographic method was developed and validated for the simultaneous determination of the β-lactamase inhibitors clavulanic acid and tazobactam in bovine milk. The HPLC system was equipped with ultraviolet absorption detection using a C18 column with a detection wavelength of 215 nm. The mobile phase (pH 4) was 0.02% phosphoric acid and methanol in the ratio of 90:10 v/v. The retention times were 5.67 min for clavulanic acid and 7.22 min for tazobactam. The method exhibited good linearity for clavulanic acid and tazobactam, with R(2) > 0.9988. Recovery ranged from 81.953% to 87.688% for clavulanic acid and from 85.007% to 92.991% for tazobactam. The precision expressed as RSD ranged from 0.975% to 1.248% for clavulanic acid and from 0.872% to 1.650% for tazobactam. A simple and precise HPLC method was developed for the determination of clavulanic acid and tazobactam in bovine milk. This method is intended for use in surveys of milk adulteration by the relevant inspection departments.

  4. Reisolation of Staphylococcus aureus from bovine milk following experimental inoculation is influenced by fat percentage and specific immunoglobulin G1 titer in milk.

    PubMed

    Boerhout, E M; Koets, A P; Vernooij, J C M; Mols-Vorstermans, T G T; Nuijten, P J M; Rutten, V P M G; Bijlsma, J J E; Eisenberg, S W F

    2016-06-01

    The associations of management parameters, herd characteristics, and individual cow factors with bovine mastitis have been subject of many studies. The present study aimed to evaluate the association between milk composition parameters, including fat, protein, lactose, urea, and specific immunoglobulin levels, at the time of experimental bacterial inoculation of the mammary gland and subsequent shedding dynamics of Staphylococcus aureus. Sixty-eight cows were experimentally infected with S. aureus and closely monitored for 3 wk. Mixed model analyses were used to determine the influence of management and herd characteristics (farm and experimental group), individual cow factors (days in milk, milk yield, and quarter position), and a challenge-related parameter (inoculation dose) in combination with either the milk components fat, protein, lactose and urea, or the S. aureus-specific antibody isotype titers at the time of bacterial inoculation, on the number of S. aureus reisolated from milk after inoculation. A positive association was observed between the milk fat percentage and the number of S. aureus reisolated from quarter milk, and a negative relationship between the S. aureus-specific IgG1 titer in milk and the number of S. aureus. These findings should be considered in the development of a vaccine against S. aureus-induced bovine mastitis. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

    PubMed

    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.

  6. Xanthine urolithiasis in a dachshund.

    PubMed

    Flegel, T; Freistadt, R; Haider, W

    1998-10-10

    Calculi were located in the kidneys, the ureters and the bladder of a two-year-old male dachshund. The yellow-greenish calculi developed as a result of impaired transformation of xanthine to uric acid resulting in an increased concentration of xanthine in the urine. The cause of the impaired catabolism of xanthine was probably a disorder of the xanthine oxidase enzyme, which catalyses the transformation of xanthine to uric acid.

  7. Comparison of anti-pathogenic activities of the human and bovine milk N-glycome: Fucosylation is a key factor.

    PubMed

    Wang, Wen-Li; Wang, Wei; Du, Ya-Min; Wu, Hong; Yu, Xiao-Bo; Ye, Ke-Ping; Li, Chun-Bao; Jung, Yong-Sam; Qian, Ying-Juan; Voglmeir, Josef; Liu, Li

    2017-11-15

    Health differences between breast- and formula-fed infants have long been apparent despite great efforts in improving the function of baby formula by adjusting the levels of various milk nutritional components. However, the N-glycome, a type of oligosaccharide decorating a diverse range of proteins, has not been extensively studied in milk regarding its biological function. In this study, the anti-pathogenic function of the enzymatically released human and bovine milk N-glycome against 5 food-borne pathogens was investigated. The human milk N-glycome showed significantly higher activity than bovine milk. After enzymatic defucosylation of human and bovine N-glycan pool, UHPLC peak shifts were observed in both suggesting heavy fucosylation of samples. Furthermore, the anti-pathogenic activity of the defulosylated N-glycome decreased significantly, and the significance of functional difference between the two almost disappeared. This result indicates the essential role of fucosylation for the anti-pathogenic function of the milk N-glycome, especially in human milk. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Fermentation characteristics and transit tolerance of probiotic Lactobacillus casei Zhang in soymilk and bovine milk during storage.

    PubMed

    Wang, J; Guo, Z; Zhang, Q; Yan, L; Chen, W; Liu, X-M; Zhang, H-P

    2009-06-01

    Lactobacillus casei Zhang is a novel strain that was screened out of koumiss collected in Inner Mongolia, and our previous research showed that L. casei Zhang has health benefits such as cholesterol-reducing and immunomodulating effects. The fermentation characteristics of L. casei Zhang in soymilk and bovine milk and the transit tolerance of L. casei Zhang in fermented milk products during refrigerated storage for 28 d were assessed. A faster decrease in pH and faster growth of L. casei Zhang during fermentation were observed in soymilk compared with bovine milk at various inoculation rates, probably because of the low pH buffering capacity of soymilk. The fermented bovine milk samples had much higher final titratable acidity (TA) values (between 0.80 and 0.93%) than the soymilk samples (between 0.40 and 0.46%). Dramatic increases in TA values in the fermented soymilk samples during storage were observed, and the TA values of the fermented soymilk samples changed from <0.56% to values between 0.86 and 0.98%. On the other hand, only slight increases in TA were observed in the bovine milk samples during the 28 d of storage. The survival rates of freshly prepared cultures of L. casei Zhang in simulated gastric juice at pH 2.0 and 2.5 were 31 and 69%, respectively, and the delivery of L. casei Zhang through fermented soymilk and bovine milk significantly improved the viability of L. casei Zhang in simulated gastric transit. Lactobacillus casei Zhang showed good tolerance to simulated gastric juice and intestinal juice in the fermented soymilk and bovine milk samples, and maintained high viability (>10(8) cfu/g) during storage at 4 degrees C for 28 d. Our results indicated that both soymilk and bovine milk could serve as vehicles for delivery of probiotic L. casei Zhang, and further research is needed to elucidate the mechanism of the change in pH and TA of L. casei Zhang in fermented milk samples during fermentation and storage and to understand the difference between

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

    PubMed

    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%.

  10. A rapid immunomagnetic beads-based immunoassay for the detection of β-casein in bovine milk.

    PubMed

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

    2014-09-01

    An immunomagnetic beads-based enzyme-linked immunosorbent assay (IMBs-ELISA) was developed for the detection of β-casein in bovine milk. Immunomagnetic beads (IMBs) were employed as the solid phase. The anti-β-casein monoclonal antibody (McAb) bound to IMBs was used as capture probe and an anti-β-casein polyclonal antibody (PcAb), labelled with horseradish peroxidase (HRP), was employed as detector probe. Three reaction and two washing steps were needed. Each reaction needed 10 min or less, which significantly shortened detection compared with classic sandwich ELISA. β-Casein in bovine milk was detected across a linear range (2-128 μg mL(-1)). Application results were in accordance with the Kjejdahl method, which suggests the IMBs-ELISA is rapid and reliable for the detection of β-casein in bovine milk.

  11. Effect of recombinant bovine somatotropin on milk production and composition of cows with Streptococcus uberis mastitis.

    PubMed

    Hoeben, D; Burvenich, C; Eppard, P J; Hard, D L

    1999-08-01

    The protective effect of bovine somatotropin (bST) during experimental Streptococcus uberis mastitis in cows was studied. The left quarters of 10 cows were infected with 500 cfu of S. uberis O140J. Five cows were subcutaneously treated with 500 mg of recombinant bST 7 d before and after infection, and 5 control cows received the excipient. In the treated cows, total milk production significantly increased after the first and second bST treatments. After infection, milk production decreased 24 and 40% in the infected quarters, 6 and 14% in the uninfected quarters, and 15 and 28% overall for treated and control cows, respectively. In the bST group, milk production was completely restored after 3 wk, but, in the control group, total production and the production of the infected quarters remained lower than preinfection production. The increase in somatic cell count occurred earlier and more rapidly in the control group, and the return to normal values was also more rapid in these cows. The amount of bacteria in milk was higher in the control cows. Changes in milk composition, such as lactose, protein, fat, Na+, K+, and Cl-, were significantly more pronounced in the control cows. Also, clinical symptoms were more prominent in the control cows. Somatotropin protected the mammary gland from excessive production losses and compositional changes during a subsequent episode of experimentally induced Streptococcus uberis mastitis and significantly improved the normalization of production and composition, which indicates a beneficial effect on the restoration of the integrity of the blood-milk barrier.

  12. Host defence related responses in bovine milk during an experimentally induced Streptococcus uberis infection

    PubMed Central

    2014-01-01

    Background Milk contains a range of proteins of moderate or low abundance that contribute to host defence. Characterisation of these proteins, the extent to which their abundance is regulated by pathogenic stimuli, and the variability of their response between and within individual animals would facilitate a better understanding of the molecular basis for this important function of milk. Results We have characterised the host defence proteins in bovine milk and their responses to intra-mammary infection by a common Gram positive mastitis pathogen, Streptococcus uberis, using a combination of 2D gel electrophoresis and GeLC mass spectrometry. In total, 68 host defence-associated proteins were identified, 18 of which have a direct antimicrobial function, 23 of which have a pathogen-recognition function, and 27 of which have a role in modulating inflammatory or immune signalling. The responsiveness of seven proteins was quantified by western blotting; validating the proteomic analyses, quantifying the within- and between animal variability of the responses, and demonstrating the complexity and specificity of the responses to this pathogen. Conclusions These data provide a foundation for understanding the role of milk in host-microbe interaction. Furthermore they provide candidate biomarkers for mastitis diagnosis, and will inform efforts to develop dairy products with improved health-promoting properties. PMID:24721702

  13. The effect of sampling technique on PCR-based bacteriological results of bovine milk samples.

    PubMed

    Hiitiö, Heidi; Simojoki, Heli; Kalmus, Piret; Holopainen, Jani; Pyörälä, Satu; Taponen, Suvi

    2016-08-01

    The aim of the study was to evaluate the effect of sampling technique on the microbiological results of bovine milk samples using multiplex real-time PCR. Comparison was made between a technique where the milk sample was taken directly from the udder cistern of the udder quarter using a needle and vacuum tube and conventional sampling. The effect of different cycle threshold (Ct) cutoff limits on the results was also tested to estimate the amount of amplified DNA in the samples. A total of 113 quarters from 53 cows were tested pairwise using both techniques, and each sample was studied with real-time PCR. Sampling from the udder cistern reduced the number of species per sample compared with conventional sampling. In conventional samples, the number of positive Staphylococcus spp. results was over twice that of samples taken with the needle technique, indicating that most of the Staphylococcus spp. originated from the teat or environmental sources. The Ct values also showed that Staphylococcus spp. were present in most samples only in low numbers. Routine use of multiplex real-time PCR in mastitis diagnostics could benefit from critical evaluation of positive Staphylococcus spp. results with Ct values between 34.0 and 37.0. Our results emphasize the importance of a careful aseptic milk sampling technique and a microbiologically positive result for a milk sample should not be automatically interpreted as an intramammary infection or mastitis. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. A new comprehensive index for discriminating adulteration in bovine raw milk.

    PubMed

    Liu, Jing; Ren, Jing; Liu, Zhen-Min; Guo, Ben-Heng

    2015-04-01

    This paper proposes a new comprehensive index, called Q, which can effectively discriminate artificial adulterated milk from unadulterated milk. Both normal and adulterated samples of bovine raw milk were analysed by Fourier transform infrared spectroscopic instrument to measure the traditional indices of quality, including fat (FAT), protein (PRO), lactose (LAC), total solids (TS), non-fat solid (NFS), freezing point (FP) and somatic cell counts (SCC). From these traditional indices, this paper elaborates a method to build the index Q. First, correlated analysis and principle component analysis were used to select parameter pairs TS-FAT and FP-LAC as predominant variables. Second, linear-regression analysis and residual analysis are applied to determine the index Q and its discriminating ranges. The verification and two-blind trial results suggested that index Q could accurately detect milk adulteration with maltodextrin and water (as low as 1.0% of adulteration proportions), and with other nine kinds of synthetic adulterants (as low as 0.5% of adulteration proportions).

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

    PubMed Central

    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

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

    PubMed

    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

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

    PubMed

    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 < 0.001 and P = 0.02, respectively). When only the predicted LTF was included in the model, the prediction of the presence of mastitis was not accurate despite a moderate correlation between SCS and LTF (r = 0.54). The specificity and the sensitivity of models were assessed using Walloon data (i.e. internal validation) and data collected from a research herd at the University of Wisconsin - Madison (i.e. 5886 Wisconsin MIR records related to 93

  18. 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.

    PubMed

    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) except for group 1 (p = 0.345). The mean VHN for the the Murrah™ buffalo milk formula, the chicken-based formula and the S-26-Promil-Gold™ formula group were not significantly different from each other (one-way ANOVA, 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.

  19. Association of metabolic acidosis with bovine milk-based human milk fortifiers.

    PubMed

    Cibulskis, C C; Armbrecht, E S

    2015-02-01

    To compare the incidence of metabolic acidosis and feeding intolerance associated with powdered or acidified liquid human milk fortifier (HMF). This retrospective study evaluated infants ⩽ 32 weeks gestational age or ⩽ 1500 g birth weight who received human milk with either powdered or acidified liquid HMF (50 consecutively born infants per group). Primary outcomes tracked were metabolic acidosis (base excess less than -4 mmol l(-1) or bicarbonate less than 18 mmol l(-1)), feeding intolerance (gastric residual > 50% feed volume, > 3 loose stools or emesis per day, abdominal tenderness or distention), necrotizing enterocolitis, late-onset infection, death, length of hospital stay and ability to remain on HMF. Demographics, feeding practices, growth parameters and laboratory data were also collected. Significantly more infants who received acidified liquid HMF developed metabolic acidosis (P < 0.001). Base excess and bicarbonate were both significantly decreased after HMF addition in the liquid HMF group (base excess P = 0.006, bicarbonate P < 0.001). More infants were switched off liquid HMF due to metabolic acidosis or feeding intolerance than those on powdered HMF (P < 0.001). Despite increased protein intake in the liquid HMF group (P = 0.009), both groups had similar enteral caloric intakes with no difference in growth rates between the two groups. There was no significant difference in any of the other primary outcomes. Infants receiving acidified liquid human milk fortifier were more likely to develop metabolic acidosis and to be switched off HMF than those who received powdered HMF. Growth in the liquid HMF group was no different than the powdered group, despite higher protein intake.

  20. Recombinant bovine somatotropin and clinical mastitis: incidence, discarded milk following therapy, and culling.

    PubMed

    Judge, L J; Erskine, R J; Bartlett, P C

    1997-12-01

    Holstein cows (n = 555) from four Michigan dairy farms were randomly assigned to receive bovine somatotropin (bST) or to serve as untreated controls. Bovine somatotropin (500 mg) was administered every 14 d beginning at 63 to 69 d of lactation and continuing until approximately 21 d prior to dry-off or until the cow was removed from the herd. Trial objectives were to determine the effect of bST on the incidence of clinical mastitis, number of days that milk was discarded because of therapy for clinical mastitis, and culling for mastitis. A total of 127 (22.9%) cases of clinical mastitis occurred during lactation. In the pretrial period (before 63 to 69 d of lactation), 42 (33.1%) cases occurred, and 85 (66.9%) cases occurred during the trial. Of the 42 pretrial cases, 57.1% occurred in control cows, and 42.9% occurred in treated cows. Of the 85 trial cases 47.1% occurred in control cows, and 52.9% occurred in treated cows. Using logistic regression, the odds ratio for the occurrence of clinical mastitis in treated cows was 1.06 (95% confidence interval = 0.62 to 1.81). The number of days that milk was discarded following therapy for clinical mastitis and the culling rate for mastitis did not differ between study groups.

  1. spa typing and enterotoxin gene profile of Staphylococcus aureus isolated from bovine raw milk in Korea

    PubMed Central

    Hwang, Sun Young; Park, Young Kyung

    2010-01-01

    Staphylococcus aureus is a major etiological pathogen of bovine mastitis, which triggers significant economic losses in dairy herds worldwide. In this study, S. aureus strains isolated from the milk of cows suffering from mastitis in Korea were investigated by spa typing and staphylococcal enterotoxin (SE) gene profiling. Forty-four S. aureus strains were isolated from 26 farms in five provinces. All isolates grouped into five clusters and two singletons based on 14 spa types. Cluster 1 and 2 isolates comprised 38.6% and 36.4% of total isolates, respectively, which were distributed in more than four provinces. SE and SE-like toxin genes were detected in 34 (77.3%) isolates and the most frequently detected SE gene profile was seg, sei, selm, seln, and selo genes (16 isolates, 36.3%), which was comparable to one of the genomic islands, Type I νSaβ. This is a first report of spa types and the prevalence of the recently described SE and SE-like toxin genes among S. aureus isolates from bovine raw milk in Korea. Two predominant spa groups were distributed widely and recently described SE and SE-like toxin genes were detected frequently. PMID:20458153

  2. Bovine leukaemia virus DNA in fresh milk and raw beef for human consumption.

    PubMed

    Olaya-Galán, N N; Corredor-Figueroa, A P; Guzmán-Garzón, T C; Ríos-Hernandez, K S; Salas-Cárdenas, S P; Patarroyo, M A; Gutierrez, M F

    2017-09-28

    Bovine leukaemia virus (BLV) is the causative agent of enzootic bovine leucosis, which has been reported worldwide. BLV has been found recently in human tissue and it could have a significant impact on human health. A possible hypothesis regarding viral entry to humans is through the consumption of infected foodstuffs. This study was aimed at detecting the presence of BLV DNA in raw beef and fresh milk for human consumption. Nested PCR directed at the BLV gag gene (272 bp) was used as a diagnostic test. PCR products were confirmed by Sanger sequencing. Forty-nine per cent of the samples proved positive for the presence of proviral DNA. This is the first study highlighting the presence of the BLV gag gene in meat products for human consumption and confirms the presence of the viral DNA in raw milk, as in previous reports. The presence of viral DNA in food products could suggest that viral particles may also be found. Further studies are needed to confirm the presence of infected viral particles, even though the present findings could represent a first approach to BLV transmission to humans through foodstuff consumption.

  3. Increased levels of LPS-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge.

    PubMed

    Bannerman, Douglas D; Paape, Max J; Hare, William R; Sohn, Eun Jung

    2003-10-01

    Several species of gram-negative bacteria, including Escherichia coli, Klebsiella pneumoniae, and various species of Enterobacter, are common mastitis pathogens. All of these bacteria are characterized by the presence of endotoxin or lipopolysaccharide (LPS) in their outer membrane. The bovine mammary gland is highly sensitive to LPS, and LPS has been implicated, in part, in the pathogenesis of gram-negative mastitis. Recognition of LPS is a key event in the innate immune response to gram-negative infection and is mediated by the accessory molecules CD14 and LPS-binding protein (LBP). The objective of the current study was to determine whether LBP levels increased in the blood and mammary gland following LPS challenge. The left and right quarters of five midlactating Holstein cows were challenged with either saline or LPS (100 microg), respectively, and milk and blood samples collected. Basal levels of plasma and milk LBP were 38 and 6 microg/ml, respectively. Plasma LBP levels increased as early as 8 h post-LPS challenge and reached maximal levels of 138 microg/ ml by 24 h. Analysis of whey samples derived from LPS-treated quarters revealed an increase in milk LBP by 12 h. Similar to plasma, maximal levels of milk LBP (34 microg/ml) were detected 24 h following the initial LPS challenge. Increments in milk LBP levels paralleled a rise in soluble CD14 (sCD14) levels and initial rises in the levels of these proteins were temporally coincident with maximal neutrophil recruitment to the inflamed gland. Because LBP and sCD14 are known to enhance LPS-induced host cell activation and to facilitate detoxification of LPS, these data are consistent with a role for these molecules in mediating mammary gland responses to LPS.

  4. Milk yield and reproductive performance of Holstein cows testing positive for bovine tuberculosis.

    PubMed

    Mellado, Miguel; Reséndiz, Dulce; Martínez, Angel Mario; de Santiago, Maria Angeles; Véliz, Francisco Gerardo; García, Jose Eduardo

    2015-08-01

    The objective of this study was to determine if high milk-yielding Holstein cows testing positive for bovine tuberculosis (bTB) are affected in their reproductive performance and milk yield. For this purpose, 1044 healthy cows and 105 bTB reactor cows were used. Tuberculosis reactor cows were from four large commercial dairy operations from the same region which were transferred from their barns to an isolated dairy facility. Cows free from this disease were placed in the same barn as the bTB reactor cows but in an isolated division and served as control animals. The analysis of variance with a general linear model for binary data showed that the reproductive performance of bTB reactors was impaired; overall pregnancy per artificial insemination differed (P < 0.05) between bTB reactor and non-reactor cows (16.9 vs. 20.7%). Cows that were TB reactors required 4.7 ± 2.9 services per pregnancy compared with 4.3 ± 2.8 for control cows (P > 0.05). The intervals between calving and conception were similar between bTB reactors (154 ± 78 days) and control animals (150 ± 80 days). Control cows tended (P = 0.08) to produce more milk than bTB reactors over a 305-day lactation (10,684 ± 1720 vs. 10,345 ± 1736; mean ± SD). Serum metabolites indicative of nutritional stress did not differ between bTB reactor and non-reactor cows. It was concluded that both reproductive performance and milk yield decreased marginally in bTB reactor cows, which explains the reluctance of milk producers to get rid of these animals.

  5. Short communication: Discrimination between retail bovine milks with different fat contents using chemometrics and fatty acid profiling.

    PubMed

    Vargas-Bello-Pérez, Einar; Toro-Mujica, Paula; Enriquez-Hidalgo, Daniel; Fellenberg, María Angélica; Gómez-Cortés, Pilar

    2017-06-01

    We used a multivariate chemometric approach to differentiate or associate retail bovine milks with different fat contents and non-dairy beverages, using fatty acid profiles and statistical analysis. We collected samples of bovine milk (whole, semi-skim, and skim; n = 62) and non-dairy beverages (n = 27), and we analyzed them using gas-liquid chromatography. Principal component analysis of the fatty acid data yielded 3 significant principal components, which accounted for 72% of the total variance in the data set. Principal component 1 was related to saturated fatty acids (C4:0, C6:0, C8:0, C12:0, C14:0, C17:0, and C18:0) and monounsaturated fatty acids (C14:1 cis-9, C16:1 cis-9, C17:1 cis-9, and C18:1 trans-11); whole milk samples were clearly differentiated from the rest using this principal component. Principal component 2 differentiated semi-skim milk samples by n-3 fatty acid content (C20:3n-3, C20:5n-3, and C22:6n-3). Principal component 3 was related to C18:2 trans-9,trans-12 and C20:4n-6, and its lower scores were observed in skim milk and non-dairy beverages. A cluster analysis yielded 3 groups: group 1 consisted of only whole milk samples, group 2 was represented mainly by semi-skim milks, and group 3 included skim milk and non-dairy beverages. Overall, the present study showed that a multivariate chemometric approach is a useful tool for differentiating or associating retail bovine milks and non-dairy beverages using their fatty acid profile. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. Effect of a prolonged-release formulation of N-methionyl bovine somatotropin (sometribove) on milk fat.

    PubMed

    Lynch, J M; Barbano, D M; Bauman, D E; Hartnell, G F; Nemeth, M A

    1992-07-01

    Nine Holstein cows were injected bi-weekly with a prolonged-release formulation of N-methionyl bST, and 9 cows were injected with excipient. Intramuscular injections began at 60 +/- 3 d postpartum and continued at 14-d intervals for the full lactation. Administration of bST increased production of milk, total fat, and all milk fat components measured. Average fatty acid composition of milk fat was not influenced by bST treatment. Stage of lactation had a large influence on production and percentage of individual fatty acids in milk fat from both bST-treated and control cows. The stage of lactation impact on the fatty acid composition of milk fat reflected changes in the relative contributions of body fat mobilization and de novo synthesis of milk fat components in response to changes in energy balance. Initiation of bST treatment caused some transient changes in milk fatty acid composition that were related to energy balance. These changes were small compared with the normal changes because of stage of lactation in all cows. Phospholipid and cholesterol content of milk also changed with stage of lactation but were not influenced by bST treatment. Melting properties of milk fat were influenced greatly by stage of lactation. Bovine somatotropin did not cause any changes in composition or physical properties of milk fat that were outside the range of normal variation.

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

    PubMed

    Gunnerud, Ulrika; Holst, Jens J; Östman, Elin; Björck, Inger

    2012-10-12

    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. 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 25 g intrinsic or added lactose, and a white wheat bread (WWB) meal was used as reference, providing 25 g starch. Post-prandial levels in plasma of glucose, insulin, incretins and amino acids were investigated at time intervals for up to 2 h. All test meals elicited lower postprandial blood glucose responses, expressed as iAUC 0-120 min compared with the WWB (P < 0.05). The insulin response was increased following all test meals, although only significantly higher after whey. Plasma amino acids were correlated to insulin and incretin secretion (iAUC 0-60 min) (P ≤ 0.05). The lowered glycemia with the test meals (iAUC 0-90 min) was inversely correlated to GLP-1 (iAUC 0-30 min) (P ≤ 0.05). 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.

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

    PubMed Central

    2012-01-01

    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 < 0.05). The insulin response was increased following all test meals, although only significantly higher after whey. Plasma amino acids were correlated to insulin and incretin secretion (iAUC 0–60 min) (P ≤ 0.05). The lowered glycemia with the test meals (iAUC 0–90 min) was inversely correlated to GLP-1 (iAUC 0–30 min) (P ≤ 0.05). 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. PMID:23057765

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

    PubMed

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

    PubMed Central

    Ao, Jinxia; Wei, Chengjie; Si, Yu; Luo, Chaochao; Lv, Wei; Lin, Ye; Cui, Yingjun; Gao, Xuejun

    2015-01-01

    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. PMID:26694361

  11. Bovine herpesvirus 1: within-herd seroprevalence and antibody levels in bulk-tank milk.

    PubMed

    Martínez, S; Yus, E; Sanjuán, M L; Camino, F; Eiras, M C; Arnaiz, I; Diéguez, F J

    2016-12-01

    The aim of the present study was to establish a relationship between the results obtained with the enzyme-linked immunosorbent assay (ELISA) technique for antibodies (against bovine herpesvirus 1) in serum and those in milk at the herd level. For this purpose, 275 samples of bulk-tank milk were analysed with glycoprotein E (gE) antibody ELISA and 207 more were analysed with glycoprotein B (gB) antibody ELISA (482 in total). All of these samples came from dairy herds whose seroprevalence was also evaluated. The results of this study were then used to analyse the sensitivity of the bulk-tankmilk test in detecting herds with a high risk of active infection (>60% seroprevalence) and its specificity in detecting those with few (<20%) or no seropositive animals. In regard to the reference test (results in blood serum), the sensitivity of the bulk-tankmilk test in detecting herds with >60% seropositive animals was 100% for both gE and gB ELISAs. The specificity figures, for gE and gB ELISAs, respectively, were 88.4% and 99.1% for infection-free herds and 72.6% and 96% for herds with <20% seroprevalence. In a quantitative approach, Pearson's correlation coefficients, reported as a measure of linear association between herd seroprevalences and transformed optical density values recorded in bulk-tank milk, were -0.63 for gE ELISA and 0.67 for gB ELISA.

  12. An alternative derivatization reaction to the determination of doramectin in bovine milk using spectrofluorimetry

    NASA Astrophysics Data System (ADS)

    Maia, Priscila M. S.; de F. Rezende, Flavia B.; Netto, Annibal D. Pereira; de C. Marques, Flávia F.

    The doramectin (DOR), which belongs to the avermectins group (AVM), has a high antiparasitic activity and so it has been widely used in food-producing animals. The DOR shows low fluorescence quantum efficiency and as a consequence, chemical derivatization reactions are necessary to produce derivatives with improved luminescent properties before its determination by fluorimetry. As the presence of this compound in food represents a risk to human health, an easy, clean and low cost derivatization reaction, which is alternative to those usually employed and that enables its spectrofluorimetric determination in milk samples, was developed. Ethanolic solutions of DOR, containing sodium hydroxide at a final concentration of 0.25 mol L-1, after 60 min of heating at 50 °C, produced fluorescent signals 1000 times higher than the original ethanolic solution. Using these optimized conditions, a linear response range that extended from 50.00 to 1000 μg L-1, with a value of (R2) equal to 0.9970, was obtained. Average recovery of DOR was 92.5 ± 1.5% (n = 3) in bovine milk fortified samples submitted to a liquid-liquid extraction at low temperature and pre concentration process, indicating the usefulness and effectiveness of the proposed method. The proposed spectrofluorimetric method is an alternative to high-performance liquid chromatography (HPLC) based methods, allowing rapid and simple detection of doramectin in milk samples.

  13. Exfoliation rate of mammary epithelial cells in milk on bovine mastitis caused by Staphylococcus aureus is associated with bacterial load.

    PubMed

    Nagasawa, Yuya; Kiku, Yoshio; Sugawara, Kazue; Tanabe, Fuyuko; Hayashi, Tomohito

    2017-09-11

    The exfoliation rate of mammary epithelial cells (MECs) in milk is affected by physiological, breeding and environmental factors. Little is known about the relationship between the MEC exfoliation into milk and mammary-infected Staphylococcus aureus (S. aureus) load on bovine mastitis caused by S. aureus. The aim of this study was to investigate the relationship between S. aureus load and the proportion of MEC exfoliation in milk using five substantial bovine mastitis models. In 64 randomly extracted milk samples from udders at 3-21 days after S. aureus infusion, there were various samples with different numbers of S. aureus counts and somatic cell counts. No significant correlations were found between the S. aureus counts and somatic cell count (r = 0.338). In contrast, a significant correlation was noted between S. aureus counts and the proportion of cytokeratin-positive cells in the milk from the infused udders (r = 0.734, P < 0.01). In conclusion, the increasing MEC exfoliation rate in milk from mastitis udders caused by S. aureus may contribute to reduced milk yield. © 2017 Japanese Society of Animal Science.

  14. Gene expression profiles of bovine mammary epithelial cells and association with milk composition traits using RNA-seq

    USDA-ARS?s Scientific Manuscript database

    In most recent years, RNA Sequencing is rapidly emerging as the major quantitative transcriptome profiling system. Elucidation of the bovine mammary gland transcriptome with RNA-seq is essential for identifying candidate genes for milk composition traits in dairy cattle. Here we used massive paralle...

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

    PubMed

    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.

  16. Short communication: Relationship between the level of bovine leukemia virus antibody and provirus in blood and milk of cows from a naturally infected herd.

    PubMed

    Jaworski, Juan P; Porta, Natalia G; Gutierrez, Geronimo; Politzki, Romina P; Álvarez, Irene; Galarza, Roxana; Abdala, Alejandro; Calvinho, Luis; Trono, Karina G

    2016-07-01

    We explored the relationship between the level of bovine leukemia virus antibodies and provirus load during natural infection. For that purpose, a set of 50 blood and milk paired samples were analyzed for the presence of bovine leukemia virus provirus and antibodies. Additionally, provirus load and antibody titers were measured and the relationship between these variables was investigated. Bovine leukemia provirus was detected in 59% of milk samples and a negative correlation was observed between the level of milk provirus load and milk antibody titers. By the consumption of raw milk, calves might be exposed to bovine leukemia virus favoring the perinatal transmission of this disease. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  17. Effect of bovine ABCG2 Y581S polymorphism on concentrations in milk of enrofloxacin and its active metabolite ciprofloxacin.

    PubMed

    Otero, J A; García-Mateos, D; de la Fuente, A; Prieto, J G; Álvarez, A I; Merino, G

    2016-07-01

    The ATP-binding cassette transporter G2 (ABCG2) is involved in the secretion of several drugs into milk. The bovine Y581S ABCG2 polymorphism increases the secretion into milk of the fluoroquinolone danofloxacin in Holstein cows. Danofloxacin and enrofloxacin are the fluoroquinolones most widely used in veterinary medicine. Both enrofloxacin (ENRO) and its active metabolite ciprofloxacin (CIPRO) reach milk at relatively high concentrations. The aim of this work was to study the effect of the bovine Y581S ABCG2 polymorphism on in vitro transport as well as on concentrations in plasma and in milk of ENRO and CIPRO. Experiments using cells overexpressing bovine ABCG2 showed the effects of ABCG2 on the transport of CIPRO, demonstrating more efficient in vitro transport of this antimicrobial by the S581 variant as compared with the Y581 variant. Animal studies administering 2.5mg/kg of ENRO subcutaneously to Y/Y 581 and Y/S 581 cows revealed that concentrations in plasma of ENRO and CIPRO were significantly lower in Y/S animals. Regardless of the genotype, the antimicrobial profile in milk after the administration of ENRO was predominantly of CIPRO. With respect to the genotype effects on the amounts of drugs present in milk, AUC0-24 values were more than 1.2 times higher in Y/S cows for ENRO and 2.2 times for CIPRO, indicating a greater capacity of Y581S to transfer these drugs into milk. These results emphasize the clinical relevance of this polymorphism as a factor affecting the concentrations in plasma and in milk of drugs of importance in veterinary medicine.

  18. Fine mapping of a quantitative trait locus for bovine milk fat composition on Bos taurus autosome 19.

    PubMed

    Bouwman, Aniek C; Visker, Marleen H P W; van Arendonk, Johana M; Bovenhuis, Henk

    2014-02-01

    A major quantitative trait locus (QTL) for milk fat content and fatty acids in both milk and adipose tissue has been detected on Bos taurus autosome 19 (BTA19) in several cattle breeds. The objective of this study was to refine the location of the QTL on BTA19 for bovine milk fat composition using a denser set of markers. Opportunities for fine mapping were provided by imputation from 50,000 genotyped single nucleotide polymorphisms (SNP) toward a high-density SNP panel with up to 777,000 SNP. The QTL region was narrowed down to a linkage disequilibrium block formed by 22 SNP covering 85,007 bp, from 51,303,322 to 51,388,329 bp on BTA19. This linkage disequilibrium block contained 2 genes: coiled-coil domain containing 57 (CCDC57) and fatty acid synthase (FASN). The gene CCDC57 is minimally characterized and has not been associated with bovine milk fat previously, but is expressed in the mammary gland. The gene FASN has been associated with bovine milk fat and fat in adipose tissue before. This gene is a likely candidate for the QTL on BTA19 because of its involvement in de novo fat synthesis. Future studies using sequence data of both CCDC57 and FASN, and eventually functional studies, will have to be pursued to assign the causal variant(s). Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  19. Determination of bovine immunoglobulin G in bovine colostrum powders, bovine milk powders, and dietary supplements containing bovine colostrum products by an automated direct immunoassay with optical biosensor: collaborative study.

    PubMed

    Gapper, Leyton

    2013-01-01

    Nine laboratories participated in an AOAC collaborative study to determine bovine immunoglobulin G (IgG) levels in selected dairy powders and dietary supplements by surface plasmon resonance (SPR) methodology. Each sample matrix was dissolved in buffer and suitably diluted to fit within the standard curve. The sample extract was injected over a surface functionalized with affinity-purified, polyclonal goat anti-bovine IgG (H+L) antibody; IgG was then detected. SPR detection was used for the direct immunoassay and quantification was made against a calibration curve prepared from bovine serum IgG. Between each standard and sample, the surface was regenerated using 10 mM glycine at pH 1.5. The samples analyzed included the likely matrixes for which the assay would find commercial use, namely, high- and low-protein-content colostrum powders, tablets containing colostrum powder, infant formula containing colostrum powder, and some IgG-containing dairy powders, i.e., milk protein isolate, whey protein concentrate, and skim milk powder. Each laboratory provided data for the study and assayed blind duplicates of seven materials. Due to gross outliers in the majority of results from one laboratory, the data from eight laboratories were used for the statistical analysis. The repeatability RSD (RSDr) values ranged from 3.2 to 7.3%, and the reproducibility RSDR values from 13.0 to 22.6%.

  20. The Core and Seasonal Microbiota of Raw Bovine Milk in Tanker Trucks and the Impact of Transfer to a Milk Processing Facility.

    PubMed

    Kable, Mary E; Srisengfa, Yanin; Laird, Miles; Zaragoza, Jose; McLeod, Jeremy; Heidenreich, Jessie; Marco, Maria L

    2016-08-23

    Currently, the bacterial composition of raw milk in tanker trucks and the outcomes of transfer and storage of that milk at commercial processing facilities are not well understood. We set out to identify the bacteria in raw milk collected for large-scale dairy product manufacturing. Raw bovine milk samples from 899 tanker trucks arriving at two dairy processors in San Joaquin Valley of California during three seasons (spring, summer, and fall) were analyzed by community 16S rRNA gene sequencing. This analysis revealed highly diverse bacterial populations, which exhibited seasonal differences. Raw milk collected in the spring contained the most diverse bacterial communities, with the highest total cell numbers and highest proportions being those of Actinobacteria Even with this complexity, a core microbiota was present, consisting of 29 taxonomic groups and high proportions of Streptococcus and Staphylococcus and unidentified members of Clostridiales Milk samples were also collected from five large-volume silos and from 13 to 25 tankers whose contents were unloaded into each of them during 2 days in the summer. Transfer of the milk to storage silos resulted in two community types. One group of silos contained a high proportion of Streptococcus spp. and was similar in that respect to the tankers that filled them. The community found in the other group of silos was distinct and dominated by Acinetobacter Overall, despite highly diverse tanker milk community structures, distinct milk bacterial communities were selected within the processing facility environment. This knowledge can inform the development of new sanitation procedures and process controls to ensure the consistent production of safe and high-quality dairy products on a global scale. Raw milk harbors diverse bacteria that are crucial determinants of the quality and safety of fluid milk and (fermented) dairy products. These bacteria enter farm milk during transport, storage, and processing. Although

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

    PubMed

    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.

  2. Changes in physical properties of bovine milk from the colostrum period to early lactation.

    PubMed

    Tsioulpas, A; Grandison, A S; Lewis, M J

    2007-11-01

    The aim of this study was to analyze individual cows' samples from the colostrum, postcolostrum, and early lactation periods to investigate how milk composition, physical properties, stability, and suitability for processing change throughout this period. Attention was paid to the first week postpartum in which the composition of bovine mammary secretion can change markedly. Properties including pH, titratable acidity, ethanol stability (ES), rennet clotting time, and casein micelle size were analyzed, together with some compositional factors such as fat, total protein, lactose, total and ionic calcium, magnesium, citrate, phosphorus, sodium, and potassium. Total Ca (36.2 mM) and free ionic Ca (2.58 mM), Mg (5.9 mM), P (32.2 mM), and Na (24.1 mM) appeared to be high on d 5 postpartum, having decreased substantially over the first 5 d; they gradually decreased thereafter. The average pH on d 5 was only 6.49, compared with 6.64 at 1 mo postpartum. Stability measurements showed that the average ES on d 5 was 70% and the rennet clotting time was 12.2 min, which were significantly lower than values at later stages. A number of milk properties including ES, pH, protein content, and Ca2+ concentration could be useful for identifying the point of transition from colostrum to the early lactation period. Knowing the composition and physical properties of colostrum and postcolostrum secretions will help establish when such milk is suitable for processing and determine the best use for that milk.

  3. Development of a real-time PCR for detection of Mycoplasma bovis in bovine milk and lung samples.

    PubMed

    Cai, Hugh Y; Bell-Rogers, Patricia; Parker, Lois; Prescott, John F

    2005-11-01

    A real-time polymerase chain reaction (PCR) assay using hybridization probes on a LightCycler platform was developed for detection of Mycoplasma bovis from individual bovine mastitis milk and pneumonic lung tissues. The detection limit was 550 colony forming units (cfu)/ml of milk and 650 cfu/25 mg of lung tissue. A panel of bovine Mycoplasma and of other bovine-origin bacteria were tested; only M. bovis strains were positive, with a melting peak of 66.6 degrees C. Mycoplasma agalactiae PG2 was also positive and could be distinguished because it had a melting peak of 63.1 degrees C. In validation testing of clinical samples, the relative sensitivity and specificity were 100% and 99.3% for individual milks and 96.6% and 100% for the lung tissue. Using M. bovis real-time PCR, the M. bovis culture-positive milk samples were estimated to contain between 5 x 10(4) and 7.7 x 10(8) cfu/ml and the M. bovis culture-positive lungs between 1 x 10(3) and 1 x 10(9) cfu/25 mg. Isolation, confirmed with the real-time PCR and colony fluorescent antibody test, showed that at the herd level, the proportion of samples positive for M. bovis isolation in mastitis milk samples submitted to the Mastitis Laboratory, Animal Health Laboratory, University of Guelph, Ontario, Canada, was 2.4% (5/201). We conclude that this probe-based real-time PCR assay is a sensitive, specific, and rapid method to identify M. bovis infection in bovine milk and pneumonic lungs.

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

    PubMed

    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.

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

    PubMed

    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

  6. Characterization of a soluble glycolipid galactosyltransferase which occurs in bovine milk.

    PubMed

    Bushway, A A; Keenan, T W

    1979-01-29

    Bovine milk was found to contain, in soluble form, an enzyme which transfers galactose from UDPgalactose to glucosylceramide. This enzyme was partially purified by the same procedure used to isolate the galactosyltransferase of lactose synthetase. The partially purified enzyme required detergents for activity, had a pH optimum of 7.2--7.3 and required Mn2+. The apparent Km calculated for glucosylceramide was 1.33 . 10(-4) M. With glucosylceramide as acceptor the product of the reaction was identified as lactosylceramide by autoradiography on thin-layer chromatograms. Lactosylceramide was also an effective acceptor for the transferase reaction but neutral glycosphingolipids or gangliosides with terminal galactose of N-acetylgalactosamine residues were ineffective or poorly effective as acceptors. Addition of alpha-lactalbumin inhibited the transferase reaction.

  7. Combination chemoprevention of hamster buccal pouch carcinogenesis by bovine milk lactoferrin and black tea polyphenols.

    PubMed

    Mohan, K V P Chandra; Letchoumy, P Vidjaya; Hara, Y; Nagini, S

    2008-03-01

    Combination chemoprevention is a promising approach for oral cancer prevention. The authors evaluated the combined chemopreventive effects of bovine milk lactoferrin (bLF) and black tea polyphenols (Polyphenon-B) in a clinically relevant in vivo model of 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. Although dietary administration of bLF and Polyphenon-B alone significantly reduced the tumor incidence, combined administration of bLF and polyphenon-B was more effective in inhibiting DMBA-induced genotoxicity and development of HBP carcinomas by modulation of carcinogen-metabolizing enzymes and cellular redox status. These results suggest that a "designer item" approach will be useful for human oral cancer prevention strategies.

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

    PubMed

    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

  9. Different inhibitory potency of febuxostat towards mammalian and bacterial xanthine oxidoreductases: insight from molecular dynamics

    PubMed Central

    Kikuchi, Hiroto; Fujisaki, Hiroshi; Furuta, Tadaomi; Okamoto, Ken; Leimkühler, Silke; Nishino, Takeshi

    2012-01-01

    Febuxostat, a drug recently approved in the US, European Union and Japan for treatment of gout, inhibits xanthine oxidoreductase (XOR)-mediated generation of uric acid during purine catabolism. It inhibits bovine milk XOR with a Ki in the picomolar-order, but we found that it is a much weaker inhibitor of Rhodobacter capsulatus XOR, even though the substrate-binding pockets of mammalian and bacterial XOR are well-conserved as regards to catalytically important residues and three-dimensional structure, and both permit the inhibitor to be accommodated in the active site, as indicated by computational docking studies. To clarify the reason for the difference of inhibitory potency towards the two XORs, we performed molecular dynamics simulations. The results indicate that differences in mobility of hydrophobic residues that do not directly interact with the substrate account for the difference in inhibitory potency. PMID:22448318

  10. Bovine respiratory syncytial virus and bovine coronavirus antibodies in bulk tank milk - risk factors and spatial analysis.

    PubMed

    Toftaker, Ingrid; Sanchez, Javier; Stokstad, Maria; Nødtvedt, Ane

    2016-10-01

    Bovine respiratory syncytial virus (BRSV) and bovine coronavirus (BCoV) are considered widespread among cattle in Norway and worldwide. This cross-sectional study was conducted based on antibody-ELISA of bulk tank milk (BTM) from 1347 herds in two neighboring counties in western Norway. The study aims were to determine the seroprevalence at herd level, to evaluate risk factors for BRSV and BCoV seropositivity, and to assess how these factors were associated with the spatial distribution of positive herds. The overall prevalence of BRSV and BCoV positive herds in the region was 46.2% and 72.2%, respectively. Isopleth maps of the prevalence risk distribution showed large differences in prevalence risk across the study area, with the highest prevalence in the northern region. Common risk factors of importance for both viruses were herd size, geographic location, and proximity to neighbors. Seropositivity for one virus was associated with increased odds of seropositivity for the other virus. Purchase of livestock was an additional risk factor for BCoV seropositivity, included in the model as in-degree, which was defined as the number of incoming movements from individual herds, through animal purchase, over a period of five years. Local dependence and the contribution of risk factors to this effect were assessed using the residuals from two logistic regression models for each virus. One model contained only the x- and y- coordinates as predictors, the other had all significant predictors included. Spatial clusters of high values of residuals were detected using the normal model of the spatial scan statistic and visualized on maps. Adjusting for the risk factors in the final models had different impact on the spatial clusters for the two viruses: For BRSV the number of clusters was reduced from six to four, for BCoV the number of clusters remained the same, however the log-likelihood ratios changed notably. This indicates that geographical differences in proximity to

  11. Bovine milk inhibits proteolytic degradation of epidermal growth factor in human gastric and duodenal lumen.

    PubMed

    Rao, R K; Baker, R D; Baker, S S

    1998-01-01

    Degradation of epidermal growth factor (EGF) in human gastric and duodenal lumen was analyzed by incubating 125I-labeled or unlabeled human recombinant EGF with human gastric or duodenal luminal fluids in vitro. Degradation of EGF was assessed by measuring the generation of acid soluble radioactivity or by reversed-phase high-performance liquid chromatography (HPLC). Incubation with gastric luminal fluids resulted in a time- and dose-dependent degradation of labeled and unlabeled EGF at pH 2.5 but not at pH 7.5. Duodenal luminal fluids, on the other hand, degraded EGF at pH 7.5 but not at pH 2.5. The rate of degradation of unlabeled EGF in gastric luminal fluids was nearly 12-fold higher than the rate of degradation of labeled EGF, whereas only a slight difference in rates of degradation of labeled and unlabeled EGF was observed in duodenal luminal fluids. High-performance liquid chromatography analysis detected three major degradation products that eluted with retention time of 17.5 min, 20.0 min, and 22.5 min that was associated with a reduction of intact EGF (retention time 23.5 min). Defatted and decaseinated supernatant of bovine milk effectively inhibited the degradation of EGF in both gastric and duodenal luminal fluids. Dietary derived protease inhibitors, such as soya bean trypsin inhibitor, lima bean trypsin inhibitor, egg white protease inhibitor, and Bowman-Birk protease inhibitor prevented EGF degradation in duodenal luminal fluids but failed to inhibit EGF degradation in gastric luminal fluids. These results suggest that bovine milk may contain specific inhibitors that protect EGF from proteolytic degradation in human gastric lumen.

  12. Dioxins and related compounds in human breast milk collected around open dumping sites in Asian developing countries: bovine milk as a potential source.

    PubMed

    Kunisue, T; Watanabe, M; Iwata, H; Subramanian, A; Monirith, I; Minh, T B; Baburajendran, R; Tana, T S; Viet, P H; Prudente, M; Tanabe, S

    2004-10-01

    In this study, concentrations of dioxins and related compounds (DRCs)--such as polychlorinated dibenzo- p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls--were found in human breast milk from women living near dumping sites of municipal waste and reference sites in India, Cambodia, Vietnam, and the Philippines during 1999 to 2000. DRCs were detected in all human breast milk samples analyzed, demonstrating that residents in these Asian developing countries have been exposed to these contaminants. In India, the concentrations of DRCs in human breast milk from women living near the investigated dumping site were notably higher than those from women living near reference sites and from women in other Asian developing countries. Toxic equivalent quantity (TEQ) levels of DRCs were comparable with or higher than those reported in the general populations of developed countries since 1990. In contrast, levels of these contaminants in human breast milk in women from Cambodia and Vietnam were not significantly different between milk from women living near the dumping and reference sites. These results indicate that significant pollution sources for DRCs are present in Indian dumping sites and that residents there have been exposed to relatively higher levels of these contaminants. TEQ levels in human breast milk from the dumping site in India tended to decrease with an increase in the number of previous deliveries by mothers, whereas no significant relationship was observed in Cambodia, Vietnam, or the Philippines. This suggests that mothers who have been exposed to relatively high levels of DRCs transfer greater amounts of these contaminants to the first infant than later ones through breast-feeding, which in turn implies that the first children of these mothers might be at higher risk from DRCs. When the residue levels of DRCs in bovine milk collected from the Indian dumping site and reference sites were examined, TEQ levels in bovine milk

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

    PubMed Central

    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 and antimicrobial therapy. Next-generation sequencing, 16 S rRNA gene quantitative real-time PCR, and aerobic culturing were applied to assess the effect of disease and antibiotic therapy on the milk microbiome. Cows diagnosed with clinical mastitis associated with Gram-negative pathogens or negative aerobic culture were randomly allocated into 5 days of Ceftiofur intramammary treatment or remained as untreated controls. Serial milk samples were collected from the affected quarter and the ipsilateral healthy quarter of the same animal. Milk from the mastitic quarter had a higher bacterial load and reduced microbial diversity compared to healthy milk. Resolution of the disease was accompanied by increases in diversity indexes and a decrease in pathogen relative abundance. Escherichia coli-associated mastitic milk samples had a remarkably distinct bacterial profile, dominated by Enterobacteriaceae, when compared to healthy milk. However, no differences were observed in culture-negative mastitis samples when compared to healthy milk. Antimicrobial treatment had no significant effect on clinical cure, bacteriological cure, pathogen clearance rate or bacterial load. PMID:27874095

  14. Glucose transporter expression differs between bovine monocyte and macrophage subsets and is influenced by milk production.

    PubMed

    Eger, M; Hussen, J; Koy, M; Dänicke, S; Schuberth, H-J; Breves, G

    2016-03-01

    The peripartal period of dairy cows is characterized by negative energy balance and higher incidences of infectious diseases such as mastitis or metritis. With the onset of lactation, milk production is prioritized and large amounts of glucose are transported into the mammary gland. Decreased overall energy availability might impair the function of monocytes acting as key innate immune cells, which give rise to macrophages and dendritic cells and link innate and adaptive immunity. Information on glucose requirements of bovine immune cells is rare. Therefore, this study aims to evaluate glucose transporter expression of the 3 bovine monocyte subsets (classical, intermediate, and nonclassical monocytes) and monocyte-derived macrophages and to identify influences of the peripartal period. Blood samples were either collected from nonpregnant healthy cows or from 16 peripartal German Holstein cows at d -14, +7, and +21 relative to parturition. Quantitative real-time PCR was applied to determine mRNA expression of glucose transporters (GLUT) 1, GLUT3, and GLUT4 in monocyte subsets and monocyte-derived macrophages. The low GLUT1 and GLUT3 expression in nonclassical monocytes was unaltered during differentiation into macrophages, whereas in classical and intermediate monocytes GLUT expression was downregulated. Alternatively activated M2 macrophages consumed more glucose compared with classically activated M1 macrophages. The GLUT4 mRNA was only detectable in unstimulated macrophages. Neither monocytes nor macrophages were insulin responsive. In the peripartum period, monocyte GLUT1 and GLUT3 expression and the GLUT3/GLUT1 ratio were negatively correlated with lactose production. The high-affinity GLUT3 transporter appears to be the predominant glucose transporter on bovine monocytes and macrophages, especially in the peripartal period when blood glucose levels decline. Glucose transporter expression in monocytes is downregulated as a function of lactose production, which

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

    PubMed

    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.

  16. Hormone variations in serum and milk of buffaloes (Bubalus bubalis) as potential indicators of treatment with recombinant bovine somatotropin.

    PubMed

    Castigliego, Lorenzo; Li, Xiao Ning; Armani, Andrea; Grifoni, Goffredo; Boselli, Carlo; Rosati, Remo; Gianfaldoni, Daniela; Guidi, Alessandra

    2011-11-01

    Recombinant bovine somatotropin (rbST) is used to increase milk yield in cows, but it has been forbidden in some countries and in the EU. However, rbST misuse represents a concern in both bovine and buffalo dairy production. A number of studies on rbST treatment have been performed on bovines, but there are few data on buffaloes. In this study, we treated eight lactating buffaloes with biweekly injections of a slow-release formulation of rbST, for five cycles of administration, and analysed total ST and insulin-like growth factor 1 (IGF-1) variations in serum and IGF-1 in milk. The aim was to assess their power as potential indicators of rbST-treatment. Blood was collected on days 2, 5, 9 and 14 of each cycle, and milk on days 2, 9 and 14 of cycles 2 and 5. Results showed an extraordinary increase in ST levels on day 2 in treated animals, followed by a rapid decrease over the following days, while a significant increase in IGF-1 was observed both in serum and in milk throughout most of each cycle. These results suggest that serum ST levels are a good indicator of treatment. However, the rapid decrease after the peak limits the useful period of sample collection.

  17. Determination of ibuprofen, ketoprofen, diclofenac and phenylbutazone in bovine milk by gas chromatography-tandem mass spectrometry.

    PubMed

    Dowling, G; Gallo, P; Fabbrocino, S; Serpe, L; Regan, L

    2008-12-01

    A method has been developed to analyse for ibuprofen (IBP), ketoprofen (KPF), diclofenac (DCF) and phenylbutazone (PBZ) residues in bovine milk. Milk samples were extracted with acetonitrile and sample extracts were purified on Isolute C(18) solid-phase extraction cartridges. Aliquots were analysed by gas chromatography-tandem mass spectrometry (GC-MS/MS). The method was validated in bovine milk, according to the criteria defined in Commission Decision 2002/657/EC. The decision limits (CCalpha were 0.59, 2.69, 0.90 and 0.70 ng ml(-1), respectively, for IBP, KPF, DCF and PBZ, and detection capabilities (CCbeta) of 1.01, 4.58, 1.54 and 1.19 ng ml(-1), respectively, were obtained. The measurement uncertainty of the method was 17.8%, 80.9%, 28.2% and 20.2% for IBP, KPF, DCF and PBZ, respectively. Fortifying bovine milk samples (n = 18) in three separate assays show the accuracy of the method to be between 104% and 112%. The precision of the method, expressed as relative standard deviations for the within-laboratory reproducibility at the three levels of fortification (5, 7.5 and 10 ng ml(-1)) was less than 8% for IBP, DCF and PBZ, respectively. Poor precision was obtained for KPF with a relative standard deviation of 28%.

  18. Characteristics of Aerococcus viridans isolated from bovine subclinical mastitis and its effect on milk SCC, yield, and composition.

    PubMed

    Sun, Meng; Gao, Jian; Ali, Tariq; Yu, Dan; Zhang, Shiyao; Khan, Saeed U; Fanning, Séamus; Han, Bo

    2017-04-01

    Aerococcus viridians (A. viridans), an environmental Gram-positive bacterium, has been documented to be associated with bovine mastitis. However, its exact role in bovine mastitis and the changes it brings about in milk characteristics are not yet known. The objectives of the current study were to describe the antibiotic resistance of A. viridans from bovine mastitis as well as the correlation between existence of this pathogen in udders and the somatic cell counts (SCC), daily milk yield, and composition of individual cow. One-year sampling for subclinical mastitis composite milk was conducted based on monthly DHI data from September 2013 to August 2014, in a commercial herd located in Beijing, China. All samples were cultured and pathogens were identified using microbiology method. A. viridians isolates were further identified by API identification system and 16S ribosomal RNA (rRNA) sequencing method. Kirby-Bauer disk diffusion method was used to test the antibiotic resistance of A. viridians against kinds of antimicrobial substance. SCC, milk yield, and milk composition data were from monthly Dairy Herd Improvement (DHI) results. Results showed that a total of 279 (16.67%) A. viridans isolates were identified from among 1674 bacterial isolates cultured from milk samples with high SCC. The incidence of mastitis caused by A. viridans was the highest (48-53%) during the summer season. Majority of the isolates were susceptible to most of antimicrobial compounds tested, especially to β-lactams, but were found to be resistant (50-90%) to aminoglycosides, sulfonamides, and tetracycline. The average SCC of the A. viridans infected cows was significantly higher (1000.0 × 10(3) cells/mL) (P < 0.01) as compared to healthy cows (72.4 × 10(3) cells/mL) and daily milk yield decreased (P > 0.05) by 1.86 kg/day. Reductions were also observed in fat content (P > 0.05), lactose (P < 0.01), and total solids (P > 0.05), whereas protein content increased

  19. Cadmium and lead in bovine milk in the mining area of the Caudal River (Spain).

    PubMed

    González-Montaña, José Ramiro; Senís, Enrique; Gutiérrez, Abner; Prieto, Felipe

    2012-07-01

    The levels of cadmium and lead in 36 raw bovine milk samples were analysed. These samples come from seven farms with a semi-extensive grazing system and were collected between the autumn of 2007 and the winter of 2008. All the farms were located in Asturias (Spain), a zone of great industrial and mining activity in the proximities of the Caudal River. The samples were collected in sterile precleaned polypropylene tubes and frozen until the analysis. After a lyophilization process, the samples were treated with nitric acid and microwave treatment. Cadmium and lead determinations were carried out using inductively coupled plasma mass spectrometry with sensibility under 0.2 ppb for liquid matrix. The lead content was found to vary from 0.71 to 16.06 μg/kg wet weight (w.w.), and the cadmium was lower than 2 μg/kg w.w. The levels of lead in milk are higher in those farms near zones of storage of mining waste depots, thermal power and areas with high levels of traffic. All the values found are in concordance with research carried out at non-polluted areas, and those for the lead are well below the European Union limitations.

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

    PubMed

    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.

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

    PubMed

    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

  2. Genetic characterization and antimicrobial resistance of Staphylococcus aureus isolated from bovine milk in Tunisia.

    PubMed

    Ben Said, M; Abbassi, M S; Bianchini, V; Sghaier, S; Cremonesi, P; Romanò, A; Gualdi, V; Hassen, A; Luini, M V

    2016-12-01

    Staphylococcus aureus is a major agent of bovine mastitis in dairy herds, causing economic losses in dairy industry worldwide. In addition, milk and milk-products contaminated by Staph. aureus can cause harmful human diseases. The aim of this study was to characterize Staph. aureus strains isolated from dairy farms in Tunisia. Bulk tank milk (n = 32) and individual cow milk (n = 130) samples were collected during the period of 2013-2014. Forty-three Staph. aureus isolates were recovered and typed by spa typing, 16S-23S rRNA intergenic spacer (RS-PCR) and multiplex PCRs for 22 virulence genes. Antimicrobial resistance was also investigated with a disc diffusion test. A selected subsample of 22 strains was additionally genotyped by multilocus sequence typing. Seventeen spa types were recovered, and t2421 (n = 10), t521 (n = 6) and t2112 (n = 5) were the most common. Fourteen different RS-PCR genotypes grouped into 11 clusters were detected in our study, with predominance of the R(VI) genotype (n = 24). Eight sequence types were identified and Clonal Complex 97, corresponding to RS-PCR cluster R, was the most common (n = 10), followed by CC1 (n = 4), CC15 (n = 3) and other four accounting for one or two strains. Different combinations of virulence genes were reported, and enterotoxin genes were present in few strains (seh, n = 4; sea, n = 2; sea and seh, n = 2; sec and sel, n = 2). The majority of strains were resistant only to penicillin; only one strain was found to be multiresistant and no methicillin-resistant Staph. aureus was demonstrated. Our study reported the isolation of CC97 from bovine milk in Tunisia for the first time and confirmed the relevance of this lineage in intramammary infection in cows. This paper describes the characteristics of Staphylococcus aureus isolated from bulk tank and individual cow milk in Tunisia. All strains were genotyped by spa typing and RS-PCR, a method based on the amplification of the 16S-23S r

  3. Genetic Variation among Staphylococcus aureus Strains from Bovine Milk and Their Relevance to Methicillin-Resistant Isolates from Humans ▿

    PubMed Central

    Hata, Eiji; Katsuda, Ken; Kobayashi, Hideki; Uchida, Ikuo; Tanaka, Kiyoshi; Eguchi, Masashi

    2010-01-01

    In genetic analysis of bovine Staphylococcus aureus isolates that are recognized as an important pathogenic bacterium in bovine mastitis, multilocus sequence typing (MLST) showed strong correlation to the results of pulsed-field gel electrophoresis, coa PCR-restriction fragment length polymorphism (RFLP), spa typing, and the coagulase serotyping method. According to MLST results, strains derived from sequence type 97 (ST97) and ST705 were suggested as not only dominant bovine S. aureus lineages in Japan but also pandemic bovine S. aureus lineages. Although both lineages seem to be distantly related to each other by phylogenetic analysis, both had common characteristics, i.e., lukM/lukF′-PV and coagulase serotype VI. These characteristics were very rare among minor bovine strains and human strains and may contribute to the host specificity of these lineages. Four methicillin-resistant S. aureus (MRSA) isolates were first confirmed from bovine milk in Japan; these isolates showed geno- and serotypes that were identical or similar to those of human MRSA isolates in Japan (ST5, staphylococcal cassette chromosome mec type II [SCCmec II], Spa type t002 or t375, and coagulase serotype II, and ST89, SCCmec IIIa, Spa type t5266, and coagulase serotype I). ST5 and ST89 are uncommon among bovine isolates in the world, whereas these STs are common among human MRSA isolates in Japan. PMID:20392913

  4. The Core and Seasonal Microbiota of Raw Bovine Milk in Tanker Trucks and the Impact of Transfer to a Milk Processing Facility

    PubMed Central

    Kable, Mary E.; Srisengfa, Yanin; Laird, Miles; Zaragoza, Jose; McLeod, Jeremy; Heidenreich, Jessie

    2016-01-01

    ABSTRACT Currently, the bacterial composition of raw milk in tanker trucks and the outcomes of transfer and storage of that milk at commercial processing facilities are not well understood. We set out to identify the bacteria in raw milk collected for large-scale dairy product manufacturing. Raw bovine milk samples from 899 tanker trucks arriving at two dairy processors in San Joaquin Valley of California during three seasons (spring, summer, and fall) were analyzed by community 16S rRNA gene sequencing. This analysis revealed highly diverse bacterial populations, which exhibited seasonal differences. Raw milk collected in the spring contained the most diverse bacterial communities, with the highest total cell numbers and highest proportions being those of Actinobacteria. Even with this complexity, a core microbiota was present, consisting of 29 taxonomic groups and high proportions of Streptococcus and Staphylococcus and unidentified members of Clostridiales. Milk samples were also collected from five large-volume silos and from 13 to 25 tankers whose contents were unloaded into each of them during 2 days in the summer. Transfer of the milk to storage silos resulted in two community types. One group of silos contained a high proportion of Streptococcus spp. and was similar in that respect to the tankers that filled them. The community found in the other group of silos was distinct and dominated by Acinetobacter. Overall, despite highly diverse tanker milk community structures, distinct milk bacterial communities were selected within the processing facility environment. This knowledge can inform the development of new sanitation procedures and process controls to ensure the consistent production of safe and high-quality dairy products on a global scale. PMID:27555305

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

    PubMed

    Foddai, Alessandro; Enøe, Claes; Krogh, Kaspar; Stockmarr, Anders; Halasa, Tariq

    2014-11-01

    A stochastic simulation model was developed to estimate the time from introduction of Bovine 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 a fixed threshold prevalence of seroconverted milking cows was reached in the herd. Different thresholds were set for each ELISA, according to previous studies. For each test, antibody detection was simulated in small (70 cows), medium (150 cows) and large (320 cows) herds. The assays included were: (1) the Danish blocking ELISA, (2) the SVANOVIR(®)BVDV-Ab ELISA, and (3) the ELISA BVD/MD p80 Institute Pourquier. The validation of the model was mainly carried out by comparing the predicted incidence of persistently infected (PI) calves and the predicted detection time, with records from a BVD infected herd. Results showed that the SVANOVIR, which was the most efficient ELISA, could detect antibodies in the BTM of a large herd 280 days (95% prediction interval: 218; 568) after a transiently infected (TI) milking cow has been introduced into the herd. The estimated time to detection after introduction of one PI calf was 111 days (44; 605). With SVANOVIR ELISA the incidence of PIs and dead born calves could be limited and the impact of the disease on the animal welfare and income of farmers (before detection) could be minimized. The results from the simulation modeling can be used to improve the current Danish BVD surveillance program in detecting early infected herds. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Comparison of the Human and Bovine Milk N-Glycome via High-Performance Microfluidic Chip Liquid Chromatography and Tandem Mass Spectrometry

    PubMed Central

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

    2012-01-01

    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 nano-flow 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

  7. DNA carryover in milk samples from routine milk recording used for PCR-based diagnosis of bovine Staphylococcus aureus mastitis.

    PubMed

    Mahmmod, Yasser S; Klaas, Ilka C; Enevoldsen, Carsten

    2017-07-01

    Real-time PCR techniques are increasingly used to detect udder pathogens from milk samples collected non-aseptically at routine milk recording. The objectives of this study were (1) to estimate the statistical associations between cycle threshold (Ct) values for Staphylococcus aureus in non-aseptically collected composite samples taken at routine milk recording from cows milked consecutively with the same milking unit and milk meter; and (2) to formulate practical and plausible guidelines for understanding the diagnostic implications of PCR testing for Staph. aureus intramammary infection at routine milk recording. The study included 4 herds with conventional milking parlors and repeatedly low Ct-values for Staph. aureus (representing a high DNA load) in bulk tank milk. Composite milk samples were collected from all cows at all milking units during routine milk recording using the Tru-Test electronic milk meter (Tru-Test Group, Auckland, New Zealand) and analyzed using the PathoProof PCR (Thermo Fisher Scientific, Vantaa, Finland) assay. Milking clock times were retrieved at each milk meter to establish the milking order of the cows at each unit. A multinomial logistic regression was applied to estimate the association between Ct-values from cows milked consecutively with the same milking unit and milk meter. The following groups were selected based on Ct-values: (1) 0-31.3, (2) 31.4-33.9, (3) 34.0-37, (4) 37.1-39.9, and (5) 40 (negative result). The association between groups from cows milked consecutively with the same milking unit and milk meter was statistically significant. Approximately 60% of cows were in Ct group 5 if the antecedent cow was also in Ct group 5, but only 20% of cows were in Ct group 5 if the antecedent cow was in Ct group 1. The probability of cows being in Ct group 1 was not markedly influenced by the group of the antecedent cow. Statistical relationships in the intermediate range gave a plausible indication of a dose-response relationship

  8. Molecular characterization of human xanthine oxidoreductase: the enzyme is grossly deficient in molybdenum and substantially deficient in iron-sulphur centres

    PubMed Central

    2005-01-01

    XOR (xanthine oxidoreductase) purified from human milk was shown to contain 0.04 atom of Mo and 0.09 molecule of molybdopterin/subunit. On the basis of UV/visible and CD spectra, the human enzyme was approx. 30% deficient in iron-sulphur centres. Mo(V) EPR showed the presence of a weak rapid signal corresponding to the enzyme of low xanthine oxidase activity and a slow signal indicating a significant content of desulpho-form. Resulphuration experiments, together with calculations based on enzymic activity and Mo content, led to an estimate of 50–60% desulpho-form. Fe/S EPR showed, in addition to the well-known Fe/S I and Fe/S II species, the presence of a third Fe/S signal, named Fe/S III, which appears to replace partially Fe/S I. Comparison is made with similarly prepared bovine milk XOR, which has approx. 15-fold higher enzymic activity and Mo content. Taken along with evidence of low Mo content in the milk of other mammals, these findings add further support to the idea that XOR protein plays a physiological role in milk (e.g. in secretion) equal in importance to its catalytic function as an enzyme. PMID:15679468

  9. Proteomic analysis to unravel the effect of heat stress on gene expression and milk synthesis in bovine mammary epithelial cells.

    PubMed

    Li, Lian; Wang, Yiru; Li, Chengmin; Wang, Genlin

    2017-08-14

    Heat stress can play a negative effect on milk yield and composition of dairy cattle, leading to immeasurable economic loss. The basic components of the mammary gland are the alveoli; these alveolar mammary epithelial cells reflect the milk producing ability of dairy cows. In this study, we exposed bovine mammary epithelial cells to heat stress and compared them to a control group using isobaric tags for relative and absolute quantitation combined with liquid chromatography coupled with tandem mass spectrometry. Compared with a control group, 104 differentially elevated proteins (>1.3-fold) and 167 decreased proteins (<0.77-fold) were identified in the heat treatment group. Gene Ontology analysis identified a majority of the differentially expressed proteins are associated in cell-substrate junction assembly, catabolic processes and metabolic processes. Some of these significantly regulated proteins were related to the synthesis and secretion of milk, such as milk protein and fat. This finding was further supported by the results obtained from the reduced β-casein expression through the system of plasminogen activator - plasminogen - plasmin and decreased fatty acid synthase could partly explain why milk fat synthesis ability of dairy cows decreased under heat stress. Our results highlight the effects of heat stress on synthesis of milk protein and fat, thus providing additional clues for further studies of heat stress on dairy milk production. © 2017 Japanese Society of Animal Science.

  10. Associations between bovine IGFBP2 polymorphisms with fertility, milk production, and metabolic status in UK dairy cows.

    PubMed

    Clempson, A M; Pollott, G E; Brickell, J S; Wathes, D C

    2012-04-01

    Insulin-like growth factor binding protein-2 (IGFBP2) is a key regulator of IGF activity that has been associated with insulin resistance and obesity. In cows, IGFBP2 mRNA expression is differentially regulated according to nutritional status in different tissues including the liver, reproductive tract, and mammary gland. This study investigated associations between single nucleotide polymorphisms (SNPs) in bovine IGFBP2 with fertility, milk production, and metabolic traits in Holstein-Friesian dairy cows. Fertility was assessed in heifers by measuring age at first service, age at first conception, and age at first calving. During the first and second lactation, the number of postpartum days for commencement of luteal activity (based on milk progesterone profiles), days to first service, days to conception, average milk production per day, 305-day milk yield, total milk yield, and total days in milk were recorded. Blood samples were taken at -1, +1, and +8 weeks relative to first and second calving for assessment of metabolic status (IGF1, insulin, beta-hydroxybutyrate, and glucose). Five novel SNPs were identified in IGFBP2, two of which had significant associations with fertility (age at conception in heifers and commencement of luteal activity) and 305-day milk yield in lactation 1. Trends of association were also observed with the peripartum metabolic status, in particular the glucose, insulin, and beta-hydroxybutyrate concentrations around second calving. These results indicate that IGFBP2 SNPs may influence tissue mobilization in dairy cows and may thus be of interest for marker assisted selection.

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

    PubMed

    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.

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

    PubMed

    Gregersen, V R; Gustavsson, F; Glantz, M; Christensen, O F; Stålhammar, H; Andrén, A; Lindmark-Månsson, H; Poulsen, N A; Larsen, L B; Paulsson, M; Bendixen, C

    2015-02-01

    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 genomic regions affecting traits related to rennet-induced gelation, the aim of this study was to identify potential candidate genes affecting these traits. Hence, rennet-induced gelation, including rennet coagulation time, gel strength, and yield stress, was measured in skim milk samples collected from 379 animals of the Swedish Red breed using low-amplitude oscillation measurements. All animals had genotypes for almost 621,000 segregating single nucleotide polymorphisms (SNP), identified using the Bovine HD SNPChip (Illumina Inc., San Diego, CA). The genome was scanned for associations, haplotypes based on SNP sets comprising highly associated SNP were inferred, and the effects of the 2 most common haplotypes within each region were analyzed using mixed models. Even though the number of animals was relatively small, a total of 21 regions were identified, with 4 regions showing association with more than one trait. A major quantitative trait locus for all traits was identified around the casein cluster explaining between 9.3 to 15.2% of the phenotypic variation of the different traits. In addition, 3 other possible candidate genes were identified; that is, UDP-N-acetyl-α-d-galactosamine:polypeptide N-acetylgalactosaminyl-transferase 1 (GALNT1), playing a role in O-glycosylation of κ-casein, and 2 cathepsins, CTSZ and CTSC, possibly involved in proteolysis of milk proteins. We have shown that other genes than the casein genes themselves may be involved in the regulation of gelation traits. However, additional analysis is needed to confirm these results. To our knowledge, this is the first study identifying quantitative trait loci affecting rennet-induced gelation of skim

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

    PubMed

    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.

  14. Genome-wide association mapping for milk fat composition and fine mapping of a QTL for de novo synthesis of milk fatty acids on bovine chromosome 13.

    PubMed

    Olsen, Hanne Gro; Knutsen, Tim Martin; Kohler, Achim; Svendsen, Morten; Gidskehaug, Lars; Grove, Harald; Nome, Torfinn; Sodeland, Marte; Sundsaasen, Kristil Kindem; Kent, Matthew Peter; Martens, Harald; Lien, Sigbjørn

    2017-02-13

    Bovine milk is widely regarded as a nutritious food source for humans, although the effects of individual fatty acids on human health is a subject of debate. Based on the assumption that genomic selection offers potential to improve milk fat composition, there is strong interest to understand more about the genetic factors that influence the biosynthesis of bovine milk and the molecular mechanisms that regulate milk fat synthesis and secretion. For this reason, the work reported here aimed at identifying genetic variants that affect milk fatty acid composition in Norwegian Red cattle. Milk fatty acid composition was predicted from the nation-wide recording scheme using Fourier transform infrared spectroscopy data and applied to estimate heritabilities for 36 individual and combined fatty acid traits. The recordings were used to generate daughter yield deviations that were first applied in a genome-wide association (GWAS) study with 17,343 markers to identify quantitative trait loci (QTL) affecting fatty acid composition, and next on high-density and sequence-level datasets to fine-map the most significant QTL on BTA13 (BTA for Bos taurus chromosome). The initial GWAS revealed 200 significant associations, with the strongest signals on BTA1, 13 and 15. The BTA13 QTL highlighted a strong functional candidate gene for de novo synthesis of short- and medium-chained saturated fatty acids; acyl-CoA synthetase short-chain family member 2. However, subsequent fine-mapping using single nucleotide polymorphisms (SNPs) from a high-density chip and variants detected by resequencing showed that the effect was more likely caused by a second nearby gene; nuclear receptor coactivator 6 (NCOA6). These findings were confirmed with results from haplotype studies. NCOA6 is a nuclear receptor that interacts with transcription factors such as PPARγ, which is a major regulator of bovine milk fat synthesis. An initial GWAS revealed a highly significant QTL for de novo-synthesized fatty

  15. Enzyme-antibody dual labeled gold nanoparticles probe for ultrasensitive detection of κ-casein in bovine milk samples.

    PubMed

    Li, Y S; Zhou, Y; Meng, X Y; Zhang, Y Y; Liu, J Q; Zhang, Y; Wang, N N; Hu, P; Lu, S Y; Ren, H L; Liu, Z S

    2014-11-15

    A dual labeled probe was synthesized by coating gold nanoparticles (AuNPs) with anti-κ-CN monoclonal antibody (McAb) and horseradish peroxidase (HRP) enzyme on their surface. The McAb was used as detector and HRP was used as label for signal amplification catalytically oxidize the substrate. AuNPs were used as bridges between the McAb and HRP. Based on the probe, an immunoassay was developed for ultrasensitive detection of κ-CN in bovine milk samples. The assay has a linear response range within 4.2-560 ng mL(-1). The limit of detection (LOD) was 4.2 ng mL(-1) which was 10 times lower than that of traditional McAb-HRP based ELISA. The recoveries of κ-CN from three brand bovine milk samples were from 95.8% to 111.0% that had a good correlation (R(2)=0.998) with those obtained by official standard Kjeldahl method. For higher sensitivity and as simple as the traditional ELISA, the developed immunoassay could provide an alternative approach for ultrasensitive detection of κ-CN in bovine milk sample.

  16. Isolation of immunoglobulin G from bovine milk whey by poly(hydroxyethyl methacrylate)-based anion-exchange cryogel.

    PubMed

    Dong, Shasha; Chen, Liang; Dai, Bin; Johnson, Wilfred; Ye, Jialei; Shen, Shaochuan; Yun, Junxian; Yao, Kejian; Lin, Dong-Qiang; Yao, Shan-Jing

    2013-08-01

    Bovine milk whey contains several bioactive proteins such as α-lactalbumin, β-lactoglobulin, and immunoglobulin G (IgG). Chromatographic separation of these proteins has received much attention in the past few years. In this work, we provide a chromatographic method for the efficient isolation of IgG from bovine milk whey using a poly(2-hydroxyethyl methacrylate)-based anion-exchange cryogel. The monolithic cryogel was prepared by grafting 2-(dimethylamino) ethyl methacrylate onto the poly(2-hydroxyethyl methacrylate)-based cryogel matrix and then employed to separate IgG under various buffer pH and salt elution conditions. The results showed that the buffer pH and the salt concentration in the step elution have remarkable influences on the purity of IgG, while the IgG recovery depended mainly on the loading volume of whey for a given cryogel bed. High purity IgG (more than 95%) was obtained using the phosphate buffer with pH of 5.8 as the running buffer and the salt solution in as the elution liquid. With suitable loading volume of whey, the maximum IgG recovery of about 94% was observed. The present separation method is thus a potential choice for the isolation of high-purity IgG from bovine milk whey.

  17. Effect of recombinant bovine somatotropin and calcium salts of long-chain fatty acids on milk from Italian buffalo.

    PubMed

    Polidori, F; Sgoifo Rossi, C A; Senatore, E M; Savoini, G; Dell'Orto, V

    1997-09-01

    Fifty-one lactating Italian river buffalo were used in an 84-d study to evaluate the effects of recombinant bovine somatotropin (bST) and Ca salts of long-chain fatty acids on productive performance. Treatments were 1) control diet, 2) the control diet plus 0.3 kg/d of added Ca salts of long-chain fatty acids, 3) the control diet plus 320 mg of recombinant bST injected every 21 d for four cycles, and 4) the control diet plus 0.3 kg/d of added Ca salts of long-chain fatty acids and 320 mg of recombinant bST administered as previously described. Administration of bST and Ca salts of long-chain fatty acids increased milk production. Milk fat percentage was not affected by treatments. The percentage of short-chain fatty acids in milk fat was reduced by the addition of Ca salts. Medium-chain, long-chain, and unsaturated fatty acids in milk fat were increased by bST treatment. Milk protein percentage was decreased by the addition of Ca salts of long-chain fatty acids. Milk casein content, as a percentage of total protein or as a percentage of true protein, was unaffected by bST. Body condition score was lowered by bST administration, but the addition of Ca salts of long-chain fatty acids reduced body condition loss in buffalo that were treated with somatotropin.

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

    PubMed

    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.

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

    PubMed Central

    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

  20. Disposition of toxic metals in the agricultural food chain. 1. Steady-state bovine milk biotransfer factors

    SciTech Connect

    Stevens, J.B. )

    1991-07-01

    Pharmacokinetic analyses were conducted on published experimental data to estimate steady-state bovine milk biotransfer factors for six metals-arsenic, cadmium, chromium, lead, mercury, and nickel. Both long-term feeding studies and bolus oral dose date were evaluated. This analysis indicated that, of the metals studied, lead and arsenic transfer to milk to the greatest extent, followed by chromium, mercury, and cadmium. The lead biotransfer factor is approximately 50 times that calculated for cadmium. Insufficient information was available to calculate an accurate biotransfer factor for nickel. Metals, as components of sludge (particularly lead, arsenic, and cadmium), were not found to produce significant elevations in analyte concentrations in milk. These findings suggest that metals contained in sludge may not be as bioavailable to dairy cows as metal salts.

  1. Monitoring the bulk milk antibody response to bovine viral diarrhea in dairy herds vaccinated with inactivated vaccines.

    PubMed

    Gonzalez, A M; Arnaiz, I; Eiras, C; Camino, F; Sanjuán, M L; Yus, E; Diéguez, F J

    2014-01-01

    This study was designed to determine long-term responses in dairy herds after vaccination with 1 of 3 inactivated bovine viral diarrhea virus (BVDV) vaccines with regard to antibodies against p80 protein in bulk tank milk samples, as detected by ELISA. In the present study, 29 dairy herds were vaccinated with Bovilis BVD (MSD Animal Health, Milton Keynes, UK), 11 with Hiprabovis Balance (Laboratorios Hipra, Amer, Spain), and 9 with Pregsure BVD (Zoetis, Florham Park, NJ). In these herds, bulk tank milk samples were collected and examined at the time of the first vaccination and every 6 mo during a 3-yr period. Samples were analyzed with a commercial ELISA test for the p80 protein of BVDV. The results demonstrated that vaccination affected the level of antibodies against p80. Hence, vaccination status should be taken into consideration when interpreting bulk tank milk antibody tests.

  2. Characterization of Methicillin-Resistant and -Susceptible Staphylococcal Isolates from Bovine Milk in Northwestern China

    PubMed Central

    Li, Longping; Zhou, Luoxiong; Wang, Lihong; Xue, Huping; Zhao, Xin

    2015-01-01

    Emergence of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS) in bovine milk is a major public health concern. The primary purpose of this research was to determine molecular genetic characteristics and antibiotic resistance of staphylococcal isolates recovered from milk of mastitic cows in the Shaanxi Province in Northwestern China. One hundred and thirteen methicillin-susceptible Staphylococcus aureus (MSSA), one mecA-positive and phenotype-positive MRSA, seven mecA- and mecC- negative but phenotype-positive MRSA and two MR-CoNS including one oxacillin-susceptible mecA-positive Staphylococcus haemolyticus (OS-MRSH) and one mecA-positive and methicillin-resistant Staphylococcus epidermidis (MRSE) isolates were recovered from 214 quarter milk samples on 4 dairy farms. All above 123 isolates were subjected to antibiotic resistance profiling. S. aureus isolates were also genotyped using the spa typing and the multilocus sequence typing (MLST). Eight MRSA and 2 MR-CoNS isolates were additionally tested for SCCmec types. Resistance was common among isolates against ampicillin or penicillin (80.5%), kanamycin (68.3%), gentamicin (67.5%), tetracycline (43.9%) and chloramphenicol (30.1%). However, no isolate was resistant to vancomycin or teicoplanin. Twenty, 29 and 58 isolates showed resistance to 1, 2 or more than 2 antibiotics, respectively. The predominant multidrug resistance profile was penicillin/ampicillin/kanamycin/gentamicin/tetracycline (46 isolates). Most S. aureus isolates belonged to spa types t524 (n = 63), t11772 (a new type, n = 31) and t4207 (n = 15). At the same time, MLST types ST71 (n = 67) and ST2738 (a new type, n = 45) were identified as dominant sequence types. The mecA-positive and phenotype-positive MRSA isolate had a composite genotype t524-ST71-SCCmecIVa, while 7 mecA-negative but phenotype-positive MRSA isolates were all t524-ST71. The OS-MRSH isolate contained a type V

  3. Structure of the O-glycopeptides isolated from bovine milk component PP3.

    PubMed

    Coddeville, B; Girardet, J M; Plancke, Y; Campagna, S; Linden, G; Spik, G

    1998-04-01

    The heat-stable acid-soluble phosphoglycoprotein component PP3 was isolated from the bovine milk proteose peptone fraction by concanavalin A affinity chromatography. Glycopeptides from the ConA-bound fraction corresponding to the component PP3 were obtained by Pronase digestion and were separated by gel filtration into high and low-molecular-mass glycopeptides. In a previous work, we have investigated the structure of the N-glycans from the high-molecular-mass glycopeptides [Girardet et al. (1995) Eur J Biochem 234: 939-46]. Here, we describe the structure of the O-glycans from the low-molecular-mass glycopeptides. By combining methylation analysis, mass spectrometry, 400 MHz 1H-NMR spectroscopy and peptide sequence analysis, we show that the low-molecular-mass fraction contains several neutral glycopeptides. A mixture of the following three glycan structures linked to the Thr86 has been identified: GalNac alpha1-O-Thr, Gal(beta1-3)GalNAc alpha1-O-Thr and Gal(beta1-4)GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc alpha1-O-Thr.

  4. Effects of beta-hydroxybutyric acid on bovine milk leukocytes function in vitro.

    PubMed

    Cerone, S I; Sansinanea, A S; García, M C

    2007-03-01

    The in vitro effect of different concentrations of beta-hydroxybutyric acid (betaHBA) on bovine milk leukocytes was examined. betaHBA level similar to those found in cows with clinical ketosis induced a significant inhibitory effect on the nitroblue tetrazolium reduction as a mean of assaying the metabolic integrity of macrophages after the phorbol-mirystate- acetate and opsonized zymosan stimulation. In the same way, the H2O2 production after stimulation with both soluble and particulate agents decreased significantly in 33 and 26%, respectively, compared with cells incubated without ketone bodies. This result suggests a possible fault in the microbicidal oxidative activity. The macrophage phagocytosis also decreased in cells treated with different betaHBA concentrations, in relation to that obtained from control cells. Neutrophils migration in agarose was determined, and the mean chemotactic response was higher when the cells were incubated with lower level or absence of ketone bodies. Considering the determined differences, we hypothesize that abnormally high levels of ketone bodies could produce a direct effect on leukocyte membranes. The induction of some modification on the receptor structure impairment the interaction ligand-receptor and this may be, in part, responsible for the higher susceptibility to local infections in mammary gland during subclinical and clinical ketosis.

  5. Detection of recombinant bovine somatotropin in milk and effect of industrial processes on its stability.

    PubMed

    Le Breton, Marie-Hélène; Beck-Henzelin, Andrea; Richoz-Payot, Janique; Rochereau-Roulet, Sandrine; Pinel, Gaud; Delatour, Thierry; Le Bizec, Bruno

    2010-07-05

    A LC-MS/MS method has been developed for the direct detection of recombinant bovine somatotropin (rbST) in milk and dairy products. The sample preparation protocol is based on a solid phase extraction step followed by precipitation with cold methanol and enzymatic digestion. The analysis is focused on the tryptic N-terminal peptide, specific of the recombinant form of the hormone and the detection is performed by LC-ESI(+)-MS/MS. This method has been validated according to the European Union criteria described in the Directive 2002/657/EC. Acceptable performances, with a decision limit (CCalpha) of 1.24 ng mL(-1) and detection capability (CCbeta) of 1.92 ng mL(-1) were obtained. Calculation of repeatability and intermediate reproducibility of the signal at 100 ng mL(-1) lead to relative standard deviations lower than 20%, showing the robustness of the method. Samples subjected to various industrial processes namely, heating, freezing, defatting, pasteurization and spray-drying were then analysed in order to determine the consequences of these treatments on the stability of the hormone. Results showed that temperature related processes, such as pasteurization and spray-drying induce a loss of the hormone up to 95%. Copyright 2010 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

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

    PubMed

    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.

  8. Genetic Variation in Vitamin B-12 Content of Bovine Milk and Its Association with SNP along the Bovine Genome

    PubMed Central

    Rutten, Marc J. M.; Bouwman, Aniek C.; Sprong, R. Corinne; van Arendonk, Johan A. M.; Visker, Marleen H. P. W.

    2013-01-01

    Vitamin B-12 (also called cobalamin) is essential for human health and current intake levels of vitamin B-12 are considered to be too low. Natural enrichment of the vitamin B-12 content in milk, an important dietary source of vitamin B-12, may help to increase vitamin B-12 intake. Natural enrichment of the milk vitamin B-12 content could be achieved through genetic selection, provided there is genetic variation between cows with respect to the vitamin B-12 content in their milk. A substantial amount of genetic variation in vitamin B-12 content was detected among raw milk samples of 544 first-lactation Dutch Holstein Friesian cows. The presence of genetic variation between animals in vitamin B-12 content in milk indicates that the genotype of the cow affects the amount of vitamin B-12 that ends up in her milk and, consequently, that the average milk vitamin B-12 content of the cow population can be increased by genetic selection. A genome-wide association study revealed significant association between 68 SNP and vitamin B-12 content in raw milk of 487 first-lactation Dutch Holstein Friesian cows. This knowledge facilitates genetic selection for milk vitamin B-12 content. It also contributes to the understanding of the biological mechanism responsible for the observed genetic variation in vitamin B-12 content in milk. None of the 68 significantly associated SNP were in or near known candidate genes involved in transport of vitamin B-12 through the gastrointestinal tract, uptake by ileum epithelial cells, export from ileal cells, transport through the blood, uptake from the blood, intracellular processing, or reabsorption by the kidneys. Probably, associations relate to genes involved in alternative pathways of well-studied processes or to genes involved in less well-studied processes such as ruminal production of vitamin B-12 or secretion of vitamin B-12 by the mammary gland. PMID:23626813

  9. Genetic variation in vitamin B-12 content of bovine milk and its association with SNP along the bovine genome.

    PubMed

    Rutten, Marc J M; Bouwman, Aniek C; Sprong, R Corinne; van Arendonk, Johan A M; Visker, Marleen H P W

    2013-01-01

    Vitamin B-12 (also called cobalamin) is essential for human health and current intake levels of vitamin B-12 are considered to be too low. Natural enrichment of the vitamin B-12 content in milk, an important dietary source of vitamin B-12, may help to increase vitamin B-12 intake. Natural enrichment of the milk vitamin B-12 content could be achieved through genetic selection, provided there is genetic variation between cows with respect to the vitamin B-12 content in their milk. A substantial amount of genetic variation in vitamin B-12 content was detected among raw milk samples of 544 first-lactation Dutch Holstein Friesian cows. The presence of genetic variation between animals in vitamin B-12 content in milk indicates that the genotype of the cow affects the amount of vitamin B-12 that ends up in her milk and, consequently, that the average milk vitamin B-12 content of the cow population can be increased by genetic selection. A genome-wide association study revealed significant association between 68 SNP and vitamin B-12 content in raw milk of 487 first-lactation Dutch Holstein Friesian cows. This knowledge facilitates genetic selection for milk vitamin B-12 content. It also contributes to the understanding of the biological mechanism responsible for the observed genetic variation in vitamin B-12 content in milk. None of the 68 significantly associated SNP were in or near known candidate genes involved in transport of vitamin B-12 through the gastrointestinal tract, uptake by ileum epithelial cells, export from ileal cells, transport through the blood, uptake from the blood, intracellular processing, or reabsorption by the kidneys. Probably, associations relate to genes involved in alternative pathways of well-studied processes or to genes involved in less well-studied processes such as ruminal production of vitamin B-12 or secretion of vitamin B-12 by the mammary gland.

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

    PubMed

    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.

  11. Nitrate and Nitrite Content of Human, Formula, Bovine, and Soy Milks: Implications for Dietary Nitrite and Nitrate Recommendations

    PubMed Central

    Hord, Norman G.; Ghannam, Janine S.; Garg, Harsha K.; Berens, Pamela D.

    2011-01-01

    Abstract Background Estimation of nitrate and nitrite concentrations of milk sources may provide insight into potential health risks and benefits of these food sources for infants, children, and adults. The World Health Organization and American Academy of Pediatrics recommends exclusive consumption of human milk for the first 6 months of life. Human milk is known to confer significant nutritional and immunological benefits for the infant. Consumption of formula, cow's, and soy milk may be used as alternatives to human milk for infants. Methods We sought to estimate potential exposure to nitrate and nitrite in human, formula, bovine, and soy milk to inform total dietary exposure estimates and recommendations. Using sensitive quantitative methodologies, nitrite and nitrate were analyzed in different samples of milk. Results Human milk concentrations of colostrum (expressed days 1–3 postpartum; n = 12), transition milk (expressed days 3–7 postpartum; n = 17), and mature milk (expressed >7 days postpartum; n = 50) were 0.08 mg/100 mL nitrite and 0.19 mg/100 mL nitrate, 0.001 mg/100 mL nitrite and 0.52 mg/100 mL nitrate, and 0.001 mg/100 mL nitrite and 0.3 mg/100 mL nitrate, respectively, revealing that the absolute amounts of these anions change as the composition of milk changes. When expressed as a percentage of the World Health Organization's Acceptable Daily Intake limits, Silk® Soy Vanilla (WhiteWave Foods, Broomfield, CO) intake could result in high nitrate intakes (104% of this standard), while intake of Bright Beginnings Soy Pediatric® formula (PBM Nutritionals, Georgia, VT) could result in the highest nitrite intakes (383% of this standard). Conclusions The temporal relationship between the provision of nitrite in human milk and the development of commensal microbiota capable of reducing dietary nitrate to nitrite supports a hypothesis that humans are adapted to provide nitrite to the gastrointestinal tract from birth

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

    PubMed

    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

  13. Comparative Growth Behaviour and Biofunctionality of Lactic Acid Bacteria During Fermentation of Soy Milk and Bovine Milk.

    PubMed

    Hati, Subrota; Patel, Nikita; Mandal, Surajit

    2017-04-29

    The study reports the growth, acidification and proteolysis of eight selected lactic acid bacteria in skim and soy milk. Angiotensin-converting enzyme inhibition and antimicrobial profiles of skim and soy milk fermented by the lactic acid bacteria were also determined. Among eight lactic cultures (S. thermophilus MD2, L. helveticus V3, L. rhamnosus NS6, L. rhamnosus NS4, L. bulgaricus NCDC 09, L. acidophilus NCDC 15, L. acidophilus NCDC 298 and L. helveticus NCDC 292) studied, L. bulgaricus NCDC 09 and S. thermophilus MD2 decreased the pH of skim and soy milk in greater extent. Acid production (i.e. titratable acidity) by L. bulgaricus NCDC 09 and L. helveticus V3 was higher than other strains. Higher viable counts were observed in S. thermophilus MD2 and L. helveticus V3. Higher proteolysis was exhibited by S. thermophilus MD2 and L. rhamnosus NS6 in both skim and soy milk. Milk fermented by S. thermophilus (MD2) exhibited highest angiotensin-converting enzyme inhibition. Antimicrobial activities of cell-free supernatant of milk fermented by S. thermophilus MD2 and L. helveticus V3 were higher. All the tested lactic acid bacteria performed better in skim milk as compared to soy milk.

  14. Short communication: Detection of stx2 and elt genes in bovine milk by using a multiplex PCR system.

    PubMed

    Nandi, R D S; Campos, A C; Puño-Sarmiento, J J; Maluta, R P; Rocha, S P D; Kobayashi, R K T; Nakazato, G

    2017-10-01

    The aim of this study was to detect 2 important toxin genes from diarrheagenic Escherichia coli (DEC) in bovine milk using a new multiplex PCR. To standardize the multiplex PCR, the stx2 and elt genes were investigated for the detection of Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC), respectively. The DNA template was prepared with a thermal procedure (boiling) and a commercial kit. Samples consisted of UHT and pasteurized milk, both skimmed, and STEC and ETEC were tested in concentrations between 10(1) and 10(9) cfu/mL. With the thermal procedure, the multiplex PCR system detected both pathotypes of E. coli at 10(9) cfu/mL in UHT and pasteurized milk. When the commercial kit was used for template preparation, STEC and ETEC could be detected at concentrations as low as 10(4) cfu/mL in UHT and pasteurized milk. Negative controls (Listeria monocytogenes, Salmonella Typhimurium, Salmonella Enteritidis, and Escherichia coli strain APEC 13) were not amplified with the multiplex PCR. These results indicate that the multiplex PCR was a rapid (less than 6 h) and efficient method to detect STEC and ETEC in milk using different methods for DNA preparation; however, the commercial kit was more sensitive than the thermal procedure. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. Mastitomics, the integrated omics of bovine milk in an experimental model of Streptococcus uberis mastitis: 3. Untargeted metabolomics.

    PubMed

    Thomas, Funmilola Clara; Mudaliar, Manikhandan; Tassi, Riccardo; McNeilly, Tom N; Burchmore, Richard; Burgess, Karl; Herzyk, Pawel; Zadoks, Ruth N; Eckersall, P David

    2016-08-16

    Intramammary infection leading to bovine mastitis is the leading disease problem affecting dairy cows and has marked effects on the milk produced by infected udder quarters. An experimental model of Streptococcus uberis mastitis has previously been investigated for clinical, immunological and pathophysiological alteration in milk, and has been the subject of peptidomic and quantitative proteomic investigation. The same sample set has now been investigated with a metabolomics approach using liquid chromatography and mass spectrometry. The analysis revealed over 3000 chromatographic peaks, of which 690 were putatively annotated with a metabolite. Hierarchical clustering analysis and principal component analysis demonstrated that metabolite changes due to S. uberis infection were maximal at 81 hours post challenge with metabolites in the milk from the resolution phase at 312 hours post challenge being closest to the pre-challenge samples. Metabolic pathway analysis revealed that the majority of the metabolites mapped to carbohydrate and nucleotide metabolism show a decreasing trend in concentration up to 81 hours post-challenge whereas an increasing trend was found in lipid metabolites and di-, tri- and tetra-peptides up to the same time point. The increase in these peptides coincides with an increase in larger peptides found in the previous peptidomic analysis and is likely to be due to protease degradation of milk proteins. Components of bile acid metabolism, linked to the FXR pathway regulating inflammation, were also increased. Metabolomic analysis of the response in milk during mastitis provides an essential component to the full understanding of the mammary gland's response to infection.

  16. Monitoring of environmental contaminants in raw bovine milk and estimates of dietary intakes of children in South Korea.

    PubMed

    Kim, Dong-Gyu; Kim, MeeKyung; Jang, Jung-Hee; Bong, Young Hoon; Kim, Jeong-Han

    2013-09-01

    Persistent organic pollutants (POPs) are ubiquitous residual contaminants in the environment and in the food chain. Milk is an important matrix for monitoring POP contamination. This study determined the concentrations of POPs including polybrominated diphenyl ethers (PBDEs), hexachlorobenzene (HCB), polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like polychlorinated biphenyls (DL-PCBs) in raw bovine milk, and the data was used to estimate dietary intakes in South Korea. The average concentrations of PBDEs, HCB, PCDD/Fs, and DL-PCBs in raw milk were 0.29 ng g(-1)fat, 0.50 ng g(-1)fat, 0.27 pg TEQ g(-1)fat, and 0.33 pg TEQ g(-1)fat, respectively. No significant relationship was found between the concentrations of analytes and the regions sampled. The dietary intakes of PBDEs from the consumption of milk was calculated to be 0.26, 0.10, and 0.05ng kg(-1)bwd(-1) for the group of ages 0-6, 7-12, and 13-19, respectively. The dietary intakes of HCB was calculated to be 0.44, 0.16, and 0.09 ng kg(-1)bwd(-1) for the group of ages 0-6, 7-12, and 13-19, respectively. The dietary intakes of PCDD/Fs and DL-PCBs was calculated to be 0.78, 0.29, and 0.16pgTEQ kg(-1)bwd(-1) for the group of ages 0-6, 7-12, and 13-19, respectively. These results indicated that the residual levels of PBDEs, HCB, PCDD/Fs, and DL-PCBs in raw bovine milk were within safe levels. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Analysis of human and bovine milk lactoferrins by Rotofor and chromatofocusing.

    PubMed

    Shimazaki, K; Kawaguchi, A; Sato, T; Ueda, Y; Tomimura, T; Shimamura, S

    1993-11-01

    1. Isoelectric points of human and bovine lactoferrins were evaluated by Rotofor and chromatofocusing analysis. 2. By Rotofor, the isoelectric value of human lactoferrin fraction was determined at 8.7 and that of bovine lactoferrin at 8.8. 3. By chromatofocusing analysis, human and bovine lactoferrins showed different elution patterns. Human lactoferrin was eluted at pH 6.8-8 and bovine lactoferrin eluted at pH 8.2-8.9.

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

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

    PubMed Central

    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

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

    PubMed

    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.

  1. Miniaturized graphene-based pipette tip extraction coupled with liquid chromatography for the determination of sulfonamide residues in bovine milk.

    PubMed

    Yan, Hongyuan; Sun, Ning; Liu, Shijia; Row, Kyung Ho; Song, Yanxue

    2014-09-01

    A miniaturized graphene-based pipette tip extraction (M-G-PTE) method coupled with liquid chromatography-ultraviolet detection was developed for rapid screening of sulfadimidine, sulfachloropyridazine, sulfamonomethoxine, and sulfachloropyrazine residues in bovine milk. Because of the large surface area and unique chemical structure of graphene, an M-G-PTE device packed with 3.0mg graphene could handle 2.0mL of milk samples at one time. This M-G-PTE device showed better adsorption performance for sulfonamides (SAs) than those packed with other adsorbents such as C18, HLB, SCX, PCX, and multiwalled carbon nanotubes. Under the optimized conditions, good linearity was obtained in the range of 0.05-6.0 μg g(-1), with a correlation coefficient (r(2)) of ⩾0.9991. The recoveries at three spiking levels ranged from 90.1% to 113.5% with relative standard deviations (RSDs) of ⩽3.9%. The proposed M-G-PTE method was simple, economical, sensitive, and produced less organic pollution. Thus, it could be applied to the rapid screening of SAs in complicated bovine milk samples. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Surface Plasmon Resonance imaging-based sensing for anti-bovine immunoglobulins detection in human milk and serum.

    PubMed

    Scarano, S; Scuffi, C; Mascini, M; Minunni, M

    2011-11-30

    Only few papers deal with Surface Plasmon Resonance imaging (SPRi) direct detection on complex matrices, limiting the biosensor application to real analytical problems. In this work a SPRi biosensor for anti-bovine IgG detection in untreated human bodily fluids, i.e. diluted human serum and milk, was developed. Enhanced levels of cow's milk antibodies in children's serum are suspected for their possible correlation with Type 1 diabetes during childhood and their detection in real samples was up to now performed by classical immunoassays based on indirect detection. The biosensor was optimised in standard samples and then in untreated human milk for anti-bovine IgG direct detection. The key novelty of the work is the evaluation of matrix effect by applying to real samples an experimental and ex ante method previously developed for SPRi signal sampling in standard solutions, called "Data Analyzer"; it punctually visualises and analyses the behaviour of receptor spots of the array, to select only spot areas with the best specific vs. unspecific signal values. In this way, benefits provide by SPRi image analysis are exploited here to quantify and minimise drawbacks due to the matrix effect, allowing to by-pass every matrix pre-treatment except dilution. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Simultaneous analysis of thirteen diuretics residues in bovine milk by ultra-performance liquid chromatography/tandem mass spectrometry.

    PubMed

    Shao, Bing; Zhang, Jing; Yang, Yi; Meng, Juan; Wu, Yongning; Duan, Hejun

    2008-11-01

    A simple, rapid, sensitive and specific method used to screen and confirm multi-class diuretics residues in whole bovine milk is described. Thirteen drugs of four different classes including carbonic anhydrase inhibitors, loop, thiazide and potassium-sparing diuretics were extracted from whole milk by acetonitrile followed by further purification with hexane. The analytes were separated using an ACQUITY UPLC BEH C18 column and detected by electrospray ionization tandem mass spectrometry (ESI-MS/MS). MS data acquisition was performed by a time-scheduled multiple reaction monitoring program, selecting two ion transitions for each target compound. The overall average recoveries based on matrix-fortified curves fortified with diuretics at three levels ranged from 80.6 to 108.8% with the coefficients of variation ranging from 2.6 to 19.7% (n = 6). The limits of quantitation (LOQs) of diuretics in bovine milk were 5.0 microg/kg for spironolactone and 0.5 microg/kg for other analytes, respectively.

  4. Characterization of an apically derived epithelial membrane glycoprotein from bovine milk, which is expressed in capillary endothelia in diverse tissues

    PubMed Central

    1985-01-01

    A glycoprotein (PAS IV) of apparent Mr 76,000 was purified from bovine milk-fat-globule membrane and partially characterized. PAS IV contained mannose, galactose, and sialic acid as principal sugars (approximately 5.3% total carbohydrate [wt/wt]) and existed in milk in at least four isoelectric variants. The glycoprotein appeared to be an integral membrane protein by several criteria. PAS IV was recovered in the detergent phase of Triton X-114 extracts of milk-fat-globule membrane at room temperature. When bound to membrane, PAS IV was resistant to digestion by a number of proteinases, although after solubilization with non-ionic detergents, the protein was readily degraded. Amino acid analysis of the purified protein revealed a high percentage of amino acids with nonpolar residues. The location of PAS IV was determined in bovine tissues by using immunofluorescence techniques. In mammary tissue, PAS IV was located on both the apical surfaces of secretory epithelial cells and endothelial cells of capillaries. This glycoprotein was also detected in endothelial cells of heart, liver, spleen, pancreas, salivary gland, and small intestine. In addition to mammary epithelial cells, PAS IV was also located in certain other epithelial cells, most notably the bronchiolar epithelial cells of lung. The potential usefulness of this protein as a specific marker of capillary endothelial cells in certain tissues is discussed. PMID:3881456

  5. Evaluation of enzyme-linked immunosorbent assay using milk samples as a potential screening test of bovine tuberculosis of dairy cows in Korea.

    PubMed

    Jeon, Bo-Young; Kim, Seung-Cheol; Je, Sungmo; Kwak, Jeongyeon; Cho, Jang-Eun; Woo, Jong-Tae; Seo, Sangkyo; Shim, Hang-Sub; Park, Byoung-Ok; Lee, Sung-Sik; Cho, Sang-Nae

    2010-06-01

    An enzyme-linked immunosorbent assay (ELISA) using bulk tank milk samples was evaluated as a screening test for bovine tuberculosis (TB), a contagious chronic disease of cattle. An ELISA with MPB70, a major antigen of Mycobacterium bovis was performed using paired sets of milk and sera samples from 33 tuberculin-positive and 43 tuberculin-negative cattle. Anti-MPB70 antibodies were detected in milk samples and there was a significant correlation between seroreactivities of milk and sera samples (R(2)=0.83). Using the tuberculin skin test as the reference test, the sensitivities of ELISA using milk and sera samples were 87.8% and 81.8%, respectively, and the specificities were 97.7% and 100%, respectively. In the screening test using bulk tank milk samples from 931 dairy herds in Whasung, Gyeonggi-do, Korea, the positive rate for anti-MPB70 antibody was 4.5% (42/931) and the tuberculin-positive rate was 2.8% (26/931). Individual milk samples (n=253) were collected from randomly selected 8 problematic and 3 negative herds (positive and negative in the screening test by MPB70 ELISA using bulk tank milk samples, respectively) and tested by MPB70 milk ELISA. In the problematic herds, positive rates were 10.5% (20/190) for anti-MPB70 antibodies in milk ELISA and 2.1% (4/190) in the tuberculin skin test. More than one dairy cows were positive by milk ELISA among the problematic herds, and all tuberculin-positive dairy cows were positive in the milk ELISA. Further, no positive cows were detected in negative herds both by milk ELISA and tuberculin skin test. These results suggest that an ELISA, using bulk tank milk samples, might be a potential efficient screening test for bovine TB of dairy cows. Copyright 2009 Elsevier Ltd. All rights reserved.

  6. Characterization of multiple forms of lactophorin isolated from bovine milk whey.

    PubMed

    Kanno, C

    1989-07-01

    A glycoprotein that reacted to the antisoluble glycoprotein of bovine milk fat globule membrane was purified from the proteose-peptone of whey and designated lactophorin. Lactophorin was separated into seven components. Lactophorin and the seven components were rich in aspartic acid, threonine, serine, glutamic acid, leucine, and lysine. The content of threonine, glycine, isoleucine, lysine, and arginine varied in each component. The ratio of fucose, mannose, galactose, N-acetylgalactosamine, N-acetylglucosamine, and sialic acid of lactophorin, which contains about 18% saccharide, were 1, 6.6, 10.3, 5.5, 9.7, and 11.6, respectively, while the respective ratio of the seven components were 1, 5 to 6, 7 to 9, 3 to 4, 6 to 8, and 4 to 12. Sialic acid content varied in each component. Protein-carbohydrate linkage was N- and o-glucoside linkage. Lactophorin consisted of seven polypeptides (I to VII) with apparent molecular weights 17,000 to 67,000. Bands I, II, VI, and VII were glycoprotein. Bands VI and VII were major and had antigenicity to anti-soluble glycoprotein, while bands I to V were minor polypeptides. Component 1 consisted of only one polypeptide (VII), whereas the components 2 to 7 contained two major (VI, VII, or both) and several minor polypeptides. The sedimentation pattern of each component was a single and almost symmetrical peak. Sedimentation coefficient was 3.79 to 5.64 S and also varied in lactophorin. The results indicate that lactophorin has multiple forms.

  7. Establishment of a 3D cell culture model of primary bovine mammary epithelial cells extracted from fresh milk.

    PubMed

    Hillreiner, Maria; Müller, Nadine I; Koch, Heiner M; Schmautz, Christiane; Küster, Bernhard; Pfaffl, Michael W; Kliem, Heike

    2017-06-22

    For the investigation of molecular processes underlying diseases of the bovine mammary gland, primary bovine mammary epithelial cells (pbMEC) are used. They are known to contribute to the innate immune system of the bovine mammary gland. The functionality of pbMEC depends on the maintenance of in vivo characteristics. So far, the optimization of pbMEC culture conditions was intended in a variety of experiments. For this purpose, most of the studies used stable cell lines or primary cells obtained from udder biopsies of slaughtered animals. By contrast, within our study, pbMEC of healthy and first lactating Brown Swiss cows were non-invasively isolated from fresh milk. The non-invasively isolated pbMEC were cultivated on the extracellular matrix-like scaffold Matrigel®. Further, they were challenged with different compositions of proliferation media, containing lactogenic hormones and/or the essential amino acid L-lysine. Changes in expression levels of genes coding for milk proteins and for components of the janus kinase/signal transducers and activators of transcription (JAK-STAT) and mTOR pathways were analyzed by RT-qPCR. The secreted proteins were analyzed by LC-MS/MS measurements. We showed for the first time the establishment of a physiologically functional 3D cell culture model of pbMEC isolated from fresh milk. This represents a primary cell culture model system, based on non-invasive cell collection, that can be used to unravel physiological processes in an unbiased manner.

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

    PubMed

    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. Copyright © 2014. Published by Elsevier B.V.

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

    PubMed

    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. High-resolution melt analysis for species identification of coagulase-negative staphylococci derived from bovine milk.

    PubMed

    Ajitkumar, Praseeda; Barkema, Herman W; Zadoks, Ruth N; Morck, Douglas W; van der Meer, Frank J U M; De Buck, Jeroen

    2013-03-01

    Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens isolated from bovine milk. In this study, we report a rapid assay for species identification of CNS using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time polymerase chain reaction amplification of 16S rRNA gene fragment, spanning the variable region V1 and V2, was performed with a resulting amplicon of 215 bp. A library of distinct melt curves of reference strains of 13 common CNS species was created using HRMA. Sequencing of 16S rRNA and rpoB genes, and, when needed, tuf gene, of 100 CNS isolates obtained from Canadian Bovine Mastitis Research Network was done to determine their species identity, allowing for subsequent evaluation of the performance of HRMA for field isolates of bovine CNS. A combination of HRMA and sequencing revealed that Staphylococcus chromogenes, S. xylosus, S. simulans, and S. sciuri had multiple genotypes, complicating their resolution by HRMA. As the 3 genotypes of S. chromogenes had distinct melt curves, the 3 distinct genotypes were employed as reference strains in a blinded trial of 156 CNS isolates to identify S. chromogenes. HRMA correctly identified all S. chromogenes isolates which were later confirmed by sequencing. Staphylococcus chromogenes (68%) was most frequently found among the CNS isolates, followed by S. haemolyticus (10%) and S. xylosus (6%). The present study revealed that HRMA of 16S rRNA gene (V1-V2) could be used as a rapid, efficient, low-cost, and minimally cumbersome technique for S. chromogenes identification, the most common CNS derived from bovine milk.

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

    PubMed

    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.

  12. Effectiveness of extruded rapeseed associated with an alfalfa protein concentrate in enhancing the bovine milk fatty acid composition.

    PubMed

    Dang Van, Q C; Bejarano, L; Mignolet, E; Coulmier, D; Froidmont, E; Larondelle, Y; Focant, M

    2011-08-01

    Linseed and rapeseed, good sources of 18:3 n-3 and cis9-18:1, respectively, have been shown to improve the bovine milk fatty acid (FA) profile. However, rapeseed, unlike linseed, has little effect on the concentration of 18:3 n-3 in milk fat. Alfalfa protein concentrate (APC), besides being a valuable protein source for milk production, contains lipids rich in 18:3 n-3. Therefore, this experiment aimed at (1) evaluating the transfer efficiency of unsaturated FA (UFA), especially 18:3 n-3, of APC to bovine milk fat, and (2) evaluating whether extruded rapeseed (ER) associated with APC is as effective as extruded linseed (EL) in enhancing the bovine milk fat composition. Six lactating Holstein cows were used in a replicated 2 × 2 Latin square design with 2 iso-energy, iso-nitrogen and iso-FA corn silage-based diets (EL and ER-APC) and two 21-d periods. Extruded linseed, as main UFA source, was included in the first diet, whereas ER, as main UFA source, and APC, as supplemental 18:3 n-3, were included in the second diet. Diets were distributed as a restricted total mixed ration. Compared with the EL diet, the ER-APC diet, where ER was associated with APC, increased milk concentration of 18:3 n-3 (1.18 vs. 1.31% of FA) and cis9-18:1 (18.35 vs. 20.01% of FA). The apparent transfer efficiency of 18:3 n-3 from diet to milk was almost twice as much for the ER-APC diet than for the EL diet (7.4 vs. 3.8% of intake). Extruded linseed accounted for 84% of 18:3 n-3 provided in the EL diet, whereas ER and APC accounted for 33 and 38% of 18:3 n-3 provided in the ER-APC diet, respectively. Because both EL and ER underwent extrusion in similar conditions, these results suggest that 18:3 n-3 of EL in the EL diet and ER in the ER-APC diet were subjected to more extensive ruminal biohydrogenation than 18:3 n-3 of APC in the ER-APC diet. This experiment shows that corn silage-based diets supplemented with ER as the main UFA source, associated with APC as supplemental 18:3 n-3, are as

  13. The relationship between size and lipid composition of the bovine milk fat globule is modulated by lactation stage.

    PubMed

    Mesilati-Stahy, Ronit; Argov-Argaman, Nurit

    2014-02-15

    The effect of lactation stage and size on the lipid composition of bovine milk fat globules (MFG) and their membranes (MFGM) was investigated. MFG were separated into six size groups (1-3 μm) from samples collected in early, mid and late lactation stages (10-250 days postpartum). Fatty acid and polar lipid composition was determined in each size group, at each lactation stage. PUFA concentration was affected by lactation stage but not MFG size. Saturated fatty acid concentration at 60 days postpartum was 10% higher in small vs. large globules. Phosphatidylcholine and cholesterol followed the same concentration pattern throughout lactation in all MFG size groups. From day 100 postpartum on, phosphatidylethanolamine concentration was constant in the large MFG, but dropped twofold in the small MFG. Results suggest distinct compositional regulation for large and small MFG at different lactation stages. Such interactions between structure, composition and lactation stage may be exploited for human milk and dairy product consumption.

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

    PubMed

    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

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

    PubMed Central

    2010-01-01

    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 < 0.05) with both milk protein and fat percentage in late lactation (P < 0.01) within the cow cohort. No association was observed between this polymorphism and either yield or composition of milk within the bull population. CXCR1-777 significantly associated (P < 0.05) with fat yield in the bull population and tended to associate (P < 0.1) with somatic cell score (SCS) in the cows genotyped. CD14-1908 A allele was found to associate with increased (P < 0.05) milk fat and protein yield and also tended to associate with increased (P < 0.1) milk yield. A SERPINA1 haplotype with superior genetic merit for milk protein yield and milk fat percentage (P < 0.05) was also identified. 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. PMID:21054834

  16. Human and bovine lactoferrins in the milk of recombinant human lactoferrin-transgenic dairy cows during lactation.

    PubMed

    Hyvönen, Paula; Suojala, Leena; Haaranen, Johanna; von Wright, Atte; Pyörälä, Satu

    2006-04-01

    Seven Friesian human lactoferrin (hLf)-transgenic primiparous dairy cows expressing recombinant hLf (rhLf) in their milk were included in the study. After calving, concentrations of rhLf and bovine LF (bLf) in the milk, somatic cell count and milk yield were determined. The concentration of rhLf was found to be constant, about 2.9 mg/mL, throughout the early lactation period of 3 months. The concentration of bLf in colostrum was higher after calving, but decreased rapidly during the first days of lactation. The mean concentration of bLf was 0.15 mg/mL, but concentrations varied between cows from 0.07 mg/mL to 0.26 mg/mL. Based on that, it may be possible to improve the non-specific host defence mechanism in the mammary gland of dairy cows by enhancing the content of rhLf in the milk.

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

    PubMed

    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). Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Residue depletion study and withdrawal period for flunixin-N-methyl glucamine in bovine milk following intravenous administration.

    PubMed

    Ngoh, Maureen A; Wislocki, Peter G; Thompson, Kenneth; Katz, Terry; Weingarten, Allan; TerHune, Terry; Hurshman, Bret

    2003-07-30

    The objective of this study was to establish a withdrawal period for flunixin in milk by quantifying 5-hydroxyflunixin, the marker residue, in bovine milk as a function of time, following intravenous treatment of lactating dairy cows with flunixin-N-methyl glucamine (Banamine or Finadyne). Lactating dairy cows were dosed on three consecutive days at 2.2 mg of flunixin free acid/kg of body weight/day. Milk was collected twice daily and assayed using a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) procedure. The method was validated at concentrations in the range 0.5-250 ppb. The concentrations for 5-hydroxyflunixin measured 12 h after the last administration of drug ranged from 1.56 to 40.6 ppb for all cows. Milk concentrations for 5-hydroxyflunixin were used to establish withdrawal periods of 36 h using guidelines established by the U.S. Food and Drug Administration/Center for Veterinary Medicine and 24 h using guidelines established by the European Medicinal Evaluation Agency/Committee on Veterinary Medicinal Products.

  19. Influence of storage and heating on protein glycation levels of processed lactose-free and regular bovine milk products.

    PubMed

    Milkovska-Stamenova, Sanja; Hoffmann, Ralf

    2017-04-15

    Thermal treatment preserves the microbiological safety of milk, but also induces Maillard reactions modifying for example proteins. The purpose of this study was evaluating the influence of consumer behaviors (storage and heating) on protein glycation degrees in bovine milk products. Lactosylation and hexosylation sites were identified in ultra-high temperature (UHT), lactose-free pasteurized, and lactose-free UHT milk (ULF) and infant formula (IF) using tandem mass spectrometry (electron transfer dissociation). Overall, 303 lactosylated and 199 hexosylated peptides were identified corresponding to 170 lactosylation (31 proteins) and 117 hexosylation sites (25 proteins). In quantitative terms, storage increased lactosylation up to fourfold in UHT and IF and hexosylation up to elevenfold in ULF and threefold in IF. These levels increased additionally twofold when the stored samples were heated (40°C). In conclusion, storage and heating appear to influence protein glycation levels in milk at similar or even higher degrees than industrial processing. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Review-Factors involving in fluctuation of trace metals concentrations in bovine milk.

    PubMed

    Perween, Rubina

    2015-05-01

    Milk makes a significant contribution to human diet through provision of macronutrients, vitamins and minerals. The exact composition of milk varies with species among domestic animals according to their neonatal needs. It is recognized that imbalance in the quantity of minerals and trace elements is a serious health hazard especially for infants. Many studies reported the fluctuation in the level of metals in milk due to the influence of several factors such as geographical and exposure to environmental pollution caused by anthropogenic activity. Amongst all sources, industries take lion's share to alter the metal content in milk. The importance of different nutritional and toxic metals in milk from different geographical areas is discussed.

  1. Magnetic coupling of the molybdenum and iron-sulphur centres in xanthine oxidase and xanthine dehydrogenases.

    PubMed Central

    Lowe, D J; Bray, R C

    1978-01-01

    Magnetic interaction between molybdenum and one of the iron-sulphur centres in milk xanthine oxidase [Lowe, Lynden-Bell & Bray (1972) Biochem. J. 130, 239-249] was studied further, with particular reference to the newly discovered Mo(V) e.p.r.(electron-paramagnetic-resonance) signal, Resting II [Lowe, Barber, Pawlik & Bray (1976) Biochem. J. 155, 81-85]. E.p.r. measurements at 35GHz near to 4.2K showed that the interaction has the same sign at all molybdenum orientations and is ferromagnetic. The predicted splitting of the e.p.r. signal from the reduced iron-sulphur centre, Fe/S I, was observed, Providing positive identification of this as the other interacting species. Chemical modification of the molybdenum environment in xanthine oxidase can change the size of the interaction severalfold, but interaction always remains approximately isotropic. The interaction in turkey liver xanthine dehydrogenase is indistinguishable from that in the oxidase. However, a bacterial xanthine dehydrogenase with different iron-sulphur centres shows rather larger interaction. Guanidinium chloride disturbs the iron-sulphur centres of the oxidase, and when this occurs there is a parallel and relatively small change in the interaction. Removal of flavin from the molecule, or raising the pH to 12.0, changes the interaction slightly without affecting the chromophores themselves. It is concluded that the Fe/S I centre and the Mo are at least 1.0nm and probably nearer 2.5nm apart, and that the conformation of the protein between them is relatively stable up to pH 12. PMID:25647

  2. The endogenous GABA bioactivity of camel, bovine, goat and human milks.

    PubMed

    Limon, Agenor; Gallegos-Perez, Jose-Luis; Reyes-Ruiz, Jorge M; Aljohi, Mohammad A; Alshanqeeti, Ali S; Miledi, Ricardo

    2014-02-15

    GABA orally administered has several beneficial effects on health, including the regulation of hyperglycaemic states in humans. Those effects are similar to the effects reported for camel milk (CMk); however, it is not known whether compounds with GABAergic activity are present in milk from camels or other species. We determined CMk free-GABA concentration by LS/MS and its bioactivity on human GABA receptors. We found that camel and goat milks have significantly more bioavailable GABA than cow and human milks and are able to activate GABAρ receptors. The relationship between GABA and taurine concentrations suggests that whole camel milk may be more efficient to activate GABAρ1 receptors than goat milk. Because GABAρ receptors are normally found in enteroendocrine cells in the lumen of the digestive tract, these results suggest that GABA in camel and goat milk may participate in GABA-modulated functions of enteroendocrine cells in the GI lumen.

  3. Occurrence of mycobacteria in bovine milk samples from both individual and collective bulk tanks at farms and informal markets in the southeast region of Sao Paulo, Brazil

    PubMed Central

    2013-01-01

    Background Mycobacterium spp. is one of the most important species of zoonotic pathogens that can be transmitted from cattle to humans. The presence of these opportunistic, pathogenic bacteria in bovine milk has emerged as a public-health concern, especially among individuals who consume raw milk and related dairy products. To address this concern, the Brazilian control and eradication program focusing on bovine tuberculosis, was established in 2001. However, bovine tuberculosis continues to afflict approximately 1,3 percent of the cattle in Brazil. In the present study, 300 samples of milk from bovine herds, obtained from both individual and collective bulk tanks and informal points of sale, were cultured on Löwenstein-Jensen and Stonebrink media. Polymerase chain reaction (PCR)-based tests and restriction-enzyme pattern analysis were then performed on the colonies exhibiting phenotypes suggestive of Mycobacterium spp., which were characterized as acid-fast bacilli. Results Of the 300 bovine milk samples that were processed, 24 were positively identified as Mycobacterium spp. Molecular identification detected 15 unique mycobacterial species: Mycobacterium bovis, M. gordonae, M. fortuitum, M. intracellulare, M. flavescens, M. duvalii, M. haemophilum, M. immunogenum, M. lentiflavum, M. mucogenicum, M. novocastrense, M. parafortuitum, M. smegmatis, M. terrae and M. vaccae. The isolation of bacteria from the various locations occurred in the following proportions: 9 percent of the individual bulk-tank samples, 7 percent of the collective bulk-tank samples and 8 percent of the informal-trade samples. No statistically significant difference was observed between the presence of Mycobacterium spp. in the three types of samples collected, the milk production profiles, the presence of veterinary assistance and the reported concerns about bovine tuberculosis prevention in the herds. Conclusion The microbiological cultures associated with PCR-based identification tests are

  4. Effect of increased milking frequency in early lactation with or without recombinant bovine somatotropin.

    PubMed

    VanBaale, M J; Ledwith, D R; Thompson, J M; Burgos, R; Collier, R J; Baumgard, L H

    2005-11-01

    Multiparous Holstein cows (n = 300) were assigned to 1 of 2 milking frequency treatments at parturition. Cows were either milked 6 times (6x) or 3 times (3x) daily to determine effects on early lactation milk yields and subsequent lactation persistency with or without use of recombinant bST (rbST). Treatments included a control group milked 3x and 3 groups milked 6x for either the first 7, 14, or 21 days in milk (DIM). Those 4 groups of cows all received rbST starting at 63 DIM. The fifth treatment group was also milked 6x for the first 21 DIM but those cows received no rbST during the entire lactation. All cows returned to 3x milking after their respective treatment periods ended. Cows milked 3x tended to produce more milk (43.2 vs. 41.5 and 41.0 +/- 1.1 kg/d) during the first 9 wk of lactation compared with cows milked 6x for 7 or 21 DIM, respectively. Group milk yields after wk 9 averaged 38.3 +/- 0.7 kg/d and did not differ among various groups assigned to an increased milking frequency in early lactation. Percentages of milk fat (3.8 +/- 0.12%) and protein (2.9 +/- 0.06%) did not differ among treatments during the first 9 wk after calving. Early lactation milk yield (41.9 +/- 1.2 kg/d) did not differ between the 2 groups of cows milked 6x for 21 DIM. However, cows subsequently administered rbST (at 63 DIM) produced more milk (38.8 vs. 34.2 +/- 0.9 kg/d) from wk 10 to 44. The number of cows sent to the hospital during the 305-d trial for mastitis (97), digestive disorders (14), respiratory issues (9), lameness (22), or retained placenta (16), were not affected by treatments (chi(2) = 0.49). Under the conditions of this commercial dairy herd in Arizona, increasing milking frequency to 6 times daily for 7 to 21 d at the start of lactation conditions did not increase milk yield nor improve lactation persistency.

  5. Effect of safflower oil, flaxseed oil, monensin, and vitamin E on concentration of conjugated linoleic acid in bovine milk fat.

    PubMed

    Bell, J A; Griinari, J M; Kennelly, J J

    2006-02-01

    Conjugated linoleic acid (CLA) refers to a mixture of conjugated octadecadienoic acids of predominantly ruminant origin. The main isomer in bovine milk fat is the cis-9, trans-11 CLA. Interest in CLA increased after the discovery of its health-promoting properties, including potent anticarcinogenic activity. Two experiments were conducted to evaluate dietary strategies aimed at increasing the concentration of CLA in bovine milk fat. Both experiments were organized as a randomized complete block design with a repeated measures treatment structure. In Experiment 1, 28 Holstein cows received either a control diet or one of 3 treatments for a period of 2 wk. The control diet consisted of 60% forage (barley silage, alfalfa silage, and alfalfa hay) and 40% concentrate on a dry matter (DM) basis, fed as a total mixed ration (TMR). The concentrate was partially replaced in the treatment groups with 24 ppm of monensin (MON), 6% of DM safflower oil (SAFF), or 6% of DM safflower oil plus 24 ppm of monensin (SAFF/M). Average cis-9, trans-11 CLA levels in milk fat after 2 wk of feeding were 0.45, 0.52, 3.36, and 5.15% of total fatty acids for control, MON, SAFF, and SAFF/M, respectively. In Experiment 2, 62 Holstein cows received either a control diet or one of 5 treatment diets for a period of 9 wk. The control diet consisted of 60% forage (barley silage, alfalfa silage, and alfalfa hay) and 40% concentrate on a DM basis, fed as a TMR. The concentrate was partially replaced in the treatment groups with 6% of DM safflower oil (SAFF), 6% of DM safflower oil plus 150 IU of vitamin E/kg of DM (SAFF/E), 6% of DM safflower oil plus 24 ppm of monensin (SAFF/M), 6% of DM safflower oil plus 24 ppm of monensin plus 150 IU of vitamin E/kg of DM (SAFF/ME), or 6% of DM flaxseed oil plus 150 IU of vitamin E/kg of DM (FLAX/E). Average cis-9, trans-11 CLA levels during the treatment period were 0.68, 4.12, 3.48, 4.55, 4.75, and 2.80% of total fatty acids for control, SAFF, SAFF/E, SAFF

  6. Bovine tuberculosis and brucellosis prevalence in cattle from selected milk cooperatives in Arsi zone, Oromia region, Ethiopia

    PubMed Central

    2013-01-01

    Background Bovine tuberculosis (BTB) and bovine brucellosis are two important milk-borne zoonoses that have been shown to be prevalent to various degrees in Ethiopian cattle. The study was carried out in four Woredas (districts) around Asella town, Arsi Zone between October 2011 and March 2012 and included 318 small-holders in 13 dairy cooperatives that marketed the delivered milk. The aims of the study were i) to assess the prevalence of the two diseases in cattle in a cross-sectional study, ii) to assess potential risk factors of BTB and brucellosis to humans as well as the knowledge-attitude-practice (KAP) among these farmers towards these diseases. Results BTB testing using the comparative intradermal skin test (CIDT) was done on 584 milking cows, out of which 417 were serologically tested for brucellosis using the Rose Bengal Plate Test and reactors confirmed with an indirect ELISA test (PrioCHECK®). The individual animal prevalence was 0.3% (95% CI 0.1% to 1.3%) for BTB, 1.7% (95% CI 0.8% to 3.5%) for brucellosis and 8.9% (95% CI 6.8% to 11.5%) for MAC (Mycobacterium avium complex). Of the 13 milk cooperatives, two had at least one positive BTB reactor and five had animals positive for brucellosis. Cross-breeds accounted for 100% and 71.4% of the BTB and brucellosis reactors respectively. For both diseases, there were prevalence variations depending on Woreda. No animal was concomitant reactor for BTB and brucellosis. Raw milk was consumed by 55.4% of the respondents. 79.2% of the respondents reported touching the afterbirth with bare hands. The latter was fed to dogs in 83% of the households. One cow among the herds of the 130 interviewees had aborted in the last 12 months. Among the interviewees, 77% stated knowing tuberculosis in general but 42 out of the 130 respondents (32.3%) did not know that BTB was transmitted by livestock. Less than half (47.7%) of the respondents knew about brucellosis. Conclusions Low prevalence of both diseases reflected the

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

    PubMed

    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

  8. Effects of recombinant methionyl bovine somatotropin (sometribove) in high producing cows milked three times daily.

    PubMed

    Jordan, D C; Aguilar, A A; Olson, J D; Bailey, C; Hartnell, G F; Madsen, K S

    1991-01-01

    Effects of daily sometribove administration on milk yield and composition, body condition score, BW, and SCC were evaluated in Holstein cows milked three times daily. Lactating cows (n =104) were assigned randomly to control or sometribove-treated (25 mg/d) groups. The experimental period was 16 wk, consisting of 2-wk pretreatment, 12-wk treatment, and 2-wk posttreatment periods. All cows were injected once daily starting at 53 to 180 d postpartum, housed in free stalls, and fed one of five total mixed rations according to milk production. Body weights were measured weekly, and body condition was scored biweekly. Milk yield was recorded daily, and weekly milk samples were analyzed for fat, protein, lactose, total solids, and SCC. Milk yield and milk protein were increased 18.8% (38.6 vs. 32.5 kg/d) and 3.3% (3.1 vs. 3.0%), respectively, whereas percentage of milk fat, lactose, SNF, SCC, and BW were unaffected by treatment. Overall average body condition scores were lower for the sometribove-treated group versus control (2.2 vs. 2.4). No apparent differences in the number of cows treated for mastitis, foot rot, displaced abomasum, or lameness were observed between treatment groups. Sometribove treatment significantly enhanced milk yield (6.1 kg/d) with no apparent negative effects on health in high producing cows milked three times per day.

  9. Net analyte signal standard addition method for simultaneous determination of sulphadiazine and trimethoprim in bovine milk and veterinary medicines.

    PubMed

    Hajian, Reza; Mousavi, Esmat; Shams, Nafiseh

    2013-06-01

    Net analyte signal standard addition method has been used for the simultaneous determination of sulphadiazine and trimethoprim by spectrophotometry in some bovine milk and veterinary medicines. The method combines the advantages of standard addition method with the net analyte signal concept which enables the extraction of information concerning a certain analyte from spectra of multi-component mixtures. This method has some advantages such as the use of a full spectrum realisation, therefore it does not require calibration and prediction step and only a few measurements require for the determination. Cloud point extraction based on the phenomenon of solubilisation used for extraction of sulphadiazine and trimethoprim in bovine milk. It is based on the induction of micellar organised media by using Triton X-100 as an extraction solvent. At the optimum conditions, the norm of NAS vectors increased linearly with concentrations in the range of 1.0-150.0 μmolL(-1) for both sulphadiazine and trimethoprim. The limits of detection (LOD) for sulphadiazine and trimethoprim were 0.86 and 0.92 μmolL(-1), respectively.

  10. Construction of an Electrochemical Sensor Based on Carbon Nanotubes/Gold Nanoparticles for Trace Determination of Amoxicillin in Bovine Milk

    PubMed Central

    Muhammad, Aliyu; Yusof, Nor Azah; Hajian, Reza; Abdullah, Jaafar

    2016-01-01

    In this work, a novel electrochemical sensor was fabricated for determination of amoxicillin in bovine milk samples by decoration of carboxylated multi-walled carbon nanotubes (MWCNTs) with gold nanoparticles (AuNPs) using ethylenediamine (en) as a cross linker (AuNPs/en-MWCNTs). The constructed nanocomposite was homogenized in dimethylformamide and drop casted on screen printed electrode. Field emission scanning electron microscopy (FESEM), energy dispersive X-Ray (EDX), X-Ray diffraction (XRD) and cyclic voltammetry were used to characterize the synthesized nanocomposites. The results show that the synthesized nanocomposites induced a remarkable synergetic effect for the oxidation of amoxicillin. Effect of some parameters, including pH, buffer, scan rate, accumulation potential, accumulation time and amount of casted nanocomposites, on the sensitivity of fabricated sensor were optimized. Under the optimum conditions, there was two linear calibration ranges from 0.2–10 µM and 10–30 µM with equations of Ipa (µA) = 2.88C (µM) + 1.2017; r = 0.9939 and Ipa (µA) = 0.88C (µM) + 22.97; r = 0.9973, respectively. The limit of detection (LOD) and limit of quantitation (LOQ) were calculated as 0.015 µM and 0.149 µM, respectively. The fabricated electrochemical sensor was successfully applied for determination of Amoxicillin in bovine milk samples and all results compared with high performance liquid chromatography (HPLC) standard method. PMID:26805829

  11. Construction of an Electrochemical Sensor Based on Carbon Nanotubes/Gold Nanoparticles for Trace Determination of Amoxicillin in Bovine Milk.

    PubMed

    Muhammad, Aliyu; Yusof, Nor Azah; Hajian, Reza; Abdullah, Jaafar

    2016-01-20

    In this work, a novel electrochemical sensor was fabricated for determination of amoxicillin in bovine milk samples by decoration of carboxylated multi-walled carbon nanotubes (MWCNTs) with gold nanoparticles (AuNPs) using ethylenediamine (en) as a cross linker (AuNPs/en-MWCNTs). The constructed nanocomposite was homogenized in dimethylformamide and drop casted on screen printed electrode. Field emission scanning electron microscopy (FESEM), energy dispersive X-Ray (EDX), X-Ray diffraction (XRD) and cyclic voltammetry were used to characterize the synthesized nanocomposites. The results show that the synthesized nanocomposites induced a remarkable synergetic effect for the oxidation of amoxicillin. Effect of some parameters, including pH, buffer, scan rate, accumulation potential, accumulation time and amount of casted nanocomposites, on the sensitivity of fabricated sensor were optimized. Under the optimum conditions, there was two linear calibration ranges from 0.2-10 µM and 10-30 µM with equations of Ipa (µA) = 2.88C (µM) + 1.2017; r = 0.9939 and Ipa (µA) = 0.88C (µM) + 22.97; r = 0.9973, respectively. The limit of detection (LOD) and limit of quantitation (LOQ) were calculated as 0.015 µM and 0.149 µM, respectively. The fabricated electrochemical sensor was successfully applied for determination of Amoxicillin in bovine milk samples and all results compared with high performance liquid chromatography (HPLC) standard method.

  12. Simultaneous determination of amoxicillin and prednisolone in bovine milk using ultra-high performance liquid chromatography tandem mass spectrometry.

    PubMed

    Li, Hui; Xia, Xi; Xue, Yanan; Tang, Shusheng; Xiao, Xilong; Li, Jiancheng; Shen, Jianzhong

    2012-07-01

    A rapid and sensitive ultra-high performance liquid chromatography tandem mass spectrometric method was developed for simultaneous quantification of amoxicillin and prednisolone in bovine milk. In this method, amoxicillin, prednisolone and the internal standards penicillin G-d(7) (for amoxicillin) and prednisolone-d(6) were extracted from bovine milk using acetonitrile. The C(18) solid phase extraction cartridges were selected for cleaning-up the extracts. The analytes were determined using a triple quadrupole mass spectrometry in positive electrospray ionization and multiple reaction monitoring mode. Calibration curves were linear over a concentration range of 2-1000 μg/kg for the analytes. The mean recoveries were 89.2-92.3% for amoxicillin and 98.7-102.3% for prednisolone. Limits of detection were 0.5 μg/kg for the analytes, and the limits of quantitation were 2 μg/kg. Decision limit (CCα) and detection capability (CCβ) have also been estimated for each analyte. The method was validated according to the Commission Decision 2002/657/EC and successfully applied to the analysis of amoxicillin and prednisolone in real samples.

  13. Validation and use of the CALUX-bioassay for the determination of dioxins and PCBs in bovine milk.

    PubMed

    Bovee, T F; Hoogenboom, L A; Hamers, A R; Traag, W A; Zuidema, T; Aarts, J M; Brouwer, A; Kuiper, H A

    1998-01-01

    There is a strong need for the development of relatively cheap and rapid bioassays for the determination of dioxins and related compounds in food. A newly developed CALUX (Chemical-Activated LUciferase gene eXpression) bioassay was tested for its possible use to determine low levels of dioxins in bovine milk. Data show that this mammalian cell-based test is very sensitive for 2,3,7,8-substituted dioxins and related PCBs, thereby reflecting the relative potencies of these compounds in comparison to TCDD (TEF-values). The limit of detection was about 50 fg of TCDD. Furthermore, the response obtained with a mixture of dioxins was additive, in accordance with the TEF-principle. Milk fat was isolated by centrifugation followed by clean-up of the fat with n-pentane, removal of the fat on a 33% H2SO4 silica column, and determination of Ah receptor agonist activity with the CALUX-bioassay. An equivalent of 67 mg fat was tested per experimental unit, resulting in a limit of quantification around 1 pg i-TEQ/g fat. To investigate the performance of the method, butter fat was cleaned and spiked with a mixture of 17 different 2,3,7,8-substituted PCDD and PCDF congeners at 1, 3, 6, 9, 12 and 15 pg TEQ/g fat, as confirmed by GC/MS. In this concentration range, the method showed a recovery of TEQs around 67% (58-87%). The reproducibility, determined in three independent series showed a CV varying between 4% and 54%, with the exception of the sample spiked at 1 pg i-TEQ (CV 97%). The repeatability determined with the sample spiked at 6 pg i-TEQ/g showed a CV of 10%. Testing of 22 bovine milk samples, taken at different sites in The Netherlands, in the CALUX-assay showed combined dioxin and dioxin-like PCB levels equivalent to 1.6 pg TCDD/g fat (range 0.2-4.6). GC/MS analysis of these samples revealed an average level of 1.7 pg i-TEQ/g fat, varying between 0.5 and 4.7 pg i-TEQ/g fat. All five samples showing a GC/MS determined dioxin content of more than 2 pg i-TEQ/g fat gave a

  14. Microbial Diversity of Bovine Mastitic Milk as Described by Pyrosequencing of Metagenomic 16s rDNA

    PubMed Central

    Oikonomou, Georgios; Machado, Vinicius Silva; Santisteban, Carlos; Schukken, Ynte Hein; Bicalho, Rodrigo Carvalho

    2012-01-01

    Dairy cow mastitis is an important disease in the dairy industry. Different microbial species have been identified as causative agents in mastitis, and are traditionally diagnosed by bacterial culture. The objective of this study was to use metagenomic pyrosequencing of bacterial 16S rRNA genes to investigate bacterial DNA diversity in milk samples of mastitic and healthy dairy cows and compare the results with those obtained by classical bacterial culture. One hundred and thirty-six milk samples were collected from cows showing signs of mastitis and used for microbiological culture. Additionally, 20 milk samples were collected from healthy quarters. Bacterial DNA was isolated from the same milk samples and the 16S rRNA genes were individually amplified and pyrosequenced. Discriminant analysis showed that the groups of samples that were most clearly different from the rest and thus easily discriminated were the normal milk samples from healthy cows and those characterised by culture as Trueperella pyogenes and Streptococcus spp. The mastitis pathogens identified by culture were generally among the most frequent organisms detected by pyrosequencing, and in some cases (Escherichia coli, Klebsiella spp. and Streptococcus uberis mastitis) the single most prevalent microorganism. Trueperella pyogenes sequences were the second most prevalent sequences in mastitis cases diagnosed as Trueperella pyogenes by culture, Streptococcus dysgalactiae sequences were the second most prevalent sequences in mastitis cases diagnosed as Streptococcus dysgalactiae by culture, and Staphyloccocus aureus sequences were the third most prevalent in mastitis cases diagnosed as Staphylococcus aureus by culture. In samples that were aerobic culture negative, pyrosequencing identified DNA of bacteria that are known to cause mastitis, DNA of bacteria that are known pathogens but have so far not been associated with mastitis, and DNA of bacteria that are currently not known to be pathogens. A

  15. Antibody content of rabbit milk and serum following inhalation or ingestion of respiratory syncytial virus and bovine serum albumin

    PubMed Central

    Peri, Barbara A.; Theodore, Christine M.; Losonsky, Genevieve A.; Fishaut, J. M.; Rothberg, R. M.; Ogra, P. L.

    1982-01-01

    The contribution of bronchus-associated or gut-associated lymphoid tissues to the development of specific immunologic reactivity in lactating mammary glands was studied by evaluating the effect of the nature of the antigen and the route of immunization on milk antibody content. Groups of pregnant rabbits were immunized with respiratory syncytial virus (RSV) and bovine serum albumin (BSA) administered i.v., per oral (p.o.) or transtracheal (i.t.) routes. The response to RSV was characterized by the regular appearance of IgA anti-RSV in the colostrum, milk, bronchial, and intestinal secretions following p.o. or i.t. immunization, but not after i.v. immunization. RSV-specific IgG appeared in the colostrum, milk, and serum regardless of the route of immunization. On the other hand, the response to BSA by all three routes of immunization was characterized by the appearance of anti-BSA in serum, colostrum and milk which was solely associated with IgG. The anti-BSA isotype did not change during the 30-day nursing period and was not affected by BSA ingestion before or during pregnancy or during nursing. If BSA was reintroduced after 20 days of nursing, a sharp rise in the Ab content of milk occurred in p.o. but not i.v. immunized dams. This increased anti-BSA was also of the IgG isotype. These observations suggest that the isotypes of specific Ab responses in the lactating mammary gland of the rabbit may be determined by the physical and chemical nature of the antigens contacted on respiratory or intestinal mucosal surfaces. PMID:7200842

  16. Determination of immunoglobulin G in bovine colostrum and milk powders, and in dietary supplements of bovine origin by protein G affinity liquid chromatography: collaborative study.

    PubMed

    Abernethy, Grant; Otter, Don; Arnold, K; Austad, J; Christiansen, S; Ferreira, I; Irvine, F; Marsh, C; Massom, L R; Otter, D; Pearce, K; Stevens, J; Szpylka, J; Vyas, P; Woollard, D; Wu, C

    2010-01-01

    An AOAC collaborative study was conducted to evaluate an affinity LC procedure for measuring immunoglobulin G (IgG) in selected dairy powders. The powders were extracted with 0.15 M sodium chloride solution and the pH was adjusted to 4.6 to precipitate caseins, which would otherwise lead to an overestimation of IgG. The analyte was then bound to a commercially available Protein G affinity cartridge and selectively eluted with a glycine buffer at pH 2.5. Detection was at 280 nm and quantification was made against a calibration curve prepared from bovine serum IgG. The samples analyzed included the likely matrixes for which this assay will find commercial use, namely, high- and low-protein-content colostrum powders, tablets containing colostrum powder, and some IgG-containing dairy powders; milk protein isolate, whey protein concentrate, and skim milk powder. Eleven laboratories provided data for the study and assayed blind duplicates of six materials. The repeatability RSD values ranged from 2.1 to 4.2% and the reproducibility RSD values ranged from 6.4 to 18.5%. The Protein G method with casein removal has adequate reproducibility for measuring IgG in colostrum-derived powders that are traded on the basis of IgG content as a colostral marker.

  17. Prevalence of Coxiella burnetii in bulk milk samples from dairy bovine, ovine, caprine, and camel herds in Iran as determined by polymerase chain reaction.

    PubMed

    Rahimi, Ebrahim; Ameri, Mehrdad; Karim, Guity; Doosti, Abbas

    2011-02-01

    Q fever is a widespread zoonosis caused by the obligate intracellular micro-organism Coxiella burnetii. The objective of this study was to determine the prevalence rate of C. burnetii in bulk milk samples from dairy bovine, ovine, caprine, and camel herds in Isfahan province, Iran. In the present study, 567 bulk milk samples from 186 dairy bovine, ovine, caprine, and camel herds were tested for C. burnetii using a nested polymerase chain reaction assay. The animals whose milk samples collected for this study were clinically healthy. In total, 8 of 247 (3.2%) bovine milk samples were positive; the positive samples originated from 6 of 90 (6.7%) dairy herds. Eight of 140 (5.7%) ovine bulk milk samples from 42 sheep breeding farms and 5 of 110 (4.5%) caprine bulk milk samples from 32 goat breeding farms were positive for C. burnetii. One of 70 (1.4%) camel bulk milk samples from 22 camel breeding farms was also positive for C. burnetii. Although no extensive prevalence study was undertaken, the results of this study indicate that clinically healthy dairy animals are important sources of C. burnetii infection in Iran. To the authors' knowledge, this study is the first report of direct identification of C. burnetii using polymerase chain reaction in bulk milk samples from dairy ovine herds in Iran and the first report of direct identification of C. burnetii in bulk milk samples from dairy camel herds. Further intensive prevalence studies on Coxiella infection and on possible risks of dairy products will be needed to elucidate the epidemiology of Q fever in Iran.

  18. Bovine milk RNases modulate pro-inflammatory responses induced by nucleic acids in cultured immune and epithelial cells.

    PubMed

    Gupta, Sandeep K; Haigh, Brendan J; Seyfert, Hans-Martin; Griffin, Frank J; Wheeler, Thomas T

    2017-03-01

    Activation of innate immune receptors by exogenous substances is crucial for the detection of microbial pathogens and a subsequent inflammatory response. The inflammatory response to microbial lipopolysaccharide via Toll-like receptor 4 (TLR4) is facilitated by soluble accessory proteins, but the role of such proteins in the activation of other pathogen recognition receptors for microbial nucleic acid is not well understood. Here we demonstrate that RNase4 and RNase5 purified from bovine milk bind to Salmonella typhimurium DNA and stimulate pro-inflammatory responses induced by nucleic acid mimetics and S. typhimurium DNA in an established mouse macrophage cell culture model, RAW264.7, as well as in primary bovine mammary epithelial cells. RNase4 and 5 also modulated pro-inflammatory signalling in response to nucleic acids in bovine peripheral blood mononuclear cells, although producing a distinct response. These results support a role for RNase4 and RNase5 in mediating inflammatory signals in both immune and epithelial cells, involving mechanisms that are cell-type specific.

  19. Investigation of the persistence of rafoxanide residues in bovine milk and fate during processing.

    PubMed

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

    2013-01-01

    Rafoxanide is an effective treatment for the control of fluke infections in animals, but it is currently not permitted for treating animals whose milk is intended for human consumption. In this study, the persistence of rafoxanide residues in milk, and their migration to dairy products, was investigated following the treatment of six lactating dairy cows with Curafluke 10% oral drench. The highest concentration of rafoxanide residues detected in the individual cows milk ranged from 249 to 627 μg kg(-1) and occurred at 2-3 days post-treatment. At 2 and 23 days post-treatment (representing high and low residue concentrations) the milk was pooled into two independent aliquots, each containing the full day's milk produced by three cows. Milk products were made from pasteurised and unpasteurised milk. Pasteurisation appeared to have little impact on the stability of the residues. Rafoxanide concentrated sixfold in the cheese (week 0) compared to the starting milk (2070 vs. 349 μg kg(-1)) but was four times lower in whey (75 μg kg(-1)). Rafoxanide residues were up to 14 times higher in butter (week 0) than in the starting milk (5468 vs. 376 μg kg(-1)). Residues were found to further concentrate in butter and cheese at longer storage and ripening times, respectively. Skim-milk powder was manufactured from skim milk, and residues were 10-fold higher than in the starting skim milk (5468 vs. 376 μg kg(-1)) despite the 185°C temperature required for the process. Rafoxanide residues were stable in this skim-milk powder when stored at ambient temperature for at least 1 year. Results showed that detectable rafoxanide residues were excreted in milk for 47 days, and concentrated in the fat-based products. The analytical ranges of the UHPLC-MS/MS method used were 1.0-200 μg kg(-1) (milk and whey) and 10-2000 μg kg(-1) (other dairy products).

  20. Direct determination of Cu and Zn in fruit juices and bovine milk by thermospray flame furnace atomic absorption spectrometry.

    PubMed

    Nascentes, Clésia C; Arruda, Marco A Z; Nogueira, Ana Rita A; Nóbrega, Joaquim A

    2004-11-15

    In the present work, thermospray flame furnace atomic absorption spectrometry (TS-FF-AAS) was employed for Cu and Zn determination in bovine milk and fruit juice samples without any pretreatment. TS-FF-AAS system was optimized and a sample volume of 300mul was injected into the carrier stream (0.014moll(-1) HNO(3) at a flow rate of 0.4mlmin(-1)), and it was introduced into a hot Ni tube. The detection limits obtained for Cu and Zn in aqueous solution were 2.2 and 0.91mugl(-1), respectively, and 3.2mugl(-1) for Cu in a medium containing water-soluble tertiary amines. The relative standard deviations varied from 2.7 to 4.2% (n=12). Sample preparation was carried out by simple dilution in water or in water-soluble tertiary amines medium. Accuracy was checked by performing addition-recovery experiments as well as by using reference materials (whole milk powder, non-fat milk powder, and infant formula). Recoveries varied from 97.7 to 105.3% for Cu and Zn. All results obtained for reference materials were in agreement with certified values at a 95% confidence level.

  1. Bacterial community profiling of milk samples as a means to understand culture-negative bovine clinical mastitis.

    PubMed

    Kuehn, Joanna S; Gorden, Patrick J; Munro, Daniel; Rong, Ruichen; Dong, Qunfeng; Plummer, Paul J; Wang, Chong; Phillips, Gregory J

    2013-01-01

    Inflammation and infection of bovine mammary glands, commonly known as mastitis, imposes significant losses each year in the dairy industry worldwide. While several different bacterial species have been identified as causative agents of mastitis, many clinical mastitis cases remain culture negative, even after enrichment for bacterial growth. To understand the basis for this increasingly common phenomenon, the composition of bacterial communities from milk samples was analyzed using culture independent pyrosequencing of amplicons of 16S ribosomal RNA genes (16S rDNA). Comparisons were made of the microbial community composition of culture negative milk samples from mastitic quarters with that of non-mastitic quarters from the same animals. Genomic DNA from culture-negative clinical and healthy quarter sample pairs was isolated, and amplicon libraries were prepared using indexed primers specific to the V1-V2 region of bacterial 16S rRNA genes and sequenced using the Roche 454 GS FLX with titanium chemistry. Evaluation of the taxonomic composition of these samples revealed significant differences in the microbiota in milk from mastitic and healthy quarters. Statistical analysis identified seven bacterial genera that may be mainly responsible for the observed microbial community differences between mastitic and healthy quarters. Collectively, these results provide evidence that cases of culture negative mastitis can be associated with bacterial species that may be present below culture detection thresholds used here. The application of culture-independent bacterial community profiling represents a powerful approach to understand long-standing questions in animal health and disease.

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

    SciTech Connect

    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.

  3. Hormone-dependent milk protein gene expression in bovine mammary explants from biopsies at different stages of pregnancy.

    PubMed

    Sheehy, Paul A; Della-Vedova, James J; Nicholas, Kevin R; Wynn, Peter C

    2004-05-01

    A method for the collection of mammary biopsies developed previously was refined and used to study the endocrine regulation of bovine milk protein gene expression. Our surgical biopsy method used real-time ultrasound imaging and epidural analgesia to enable recovery of a sufficient quantity of mammary tissue from late-pregnant dairy cows for explant culture in vitro. The time of biopsy was critical for prolactin-dependent induction of milk protein gene expression in mammary explants, as only mammary tissue from cows nearing 30 d prepartum was hormone-responsive. This suggests that during the later stages of pregnancy a change in the responsiveness of milk protein gene expression to endocrine stimuli occurred in preparation for lactation. This may relate to the diminution of a putative population of undifferentiated cells that were still responsive to prolactin. Alternatively, the metabolic activity of the tissue had increased to the level whereby the response of the tissue was no longer assessable using this model in vitro.

  4. Bulk tank milk somatic cell counts in dairy herds with different bovine viral diarrhoea virus status in Poland.

    PubMed

    Rola, Jolanta G; Larska, Magdalena; Grzeszuk, Monika; Bocian, Lukasz; Kuta, Aleksandra; Polak, Miroslaw P; Rola, Jerzy

    2014-09-01

    The aim of the study was to examine the effect of bovine viral diarrhoea virus (BVDV) infection on bulk tank milk somatic cell counts (BMSCC). Twenty nine dairy farms supplying milk to a dairy in Eastern Poland were recruited for the study. Bulk milk ELISA and RT-PCR were used to determine the BVDV infection status and the presence of PI animals in the farms. The BMSCC mean values for the BVDV seronegative (218.7 × 10(3)cells/ml; SD: 89.8) and seropositive (214.9 × 10(3)cells/ml; SD: 74.0) herds did not differ significantly. To assess the relationship between BVDV infection and BMSCC a multilevel mixed-effects linear model was used. No statistically significant effect of BVDV infection on BMSCC was found. The mean values of BMSCC for the herds with PI individuals measured before (230.1 × 10(3)cells/ml, SD: 64.9) and after (223.3 × 10(3)cells/ml, SD: 62.4) the PI removal were not statistically different. An increase in herd size was associated with a significant decrease in BMSCC. An increase in BMSCC was observed during summer (from May to September) compared to during winter (from October to April).

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  6. Using metatranscriptomics to determine effects of dietary supplementation with bovine milk oligosaccharides in healthy adults

    USDA-ARS?s Scientific Manuscript database

    The study of human milk has revealed the presence of complex human milk oligosaccharide (HMO) molecules, believed to help encourage formation and development of the infant’s gut microbiome. HMOs function as prebiotics for beneficial bacteria, block attachment to pathogens, and interact directly with...

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

    PubMed Central

    Nekouei, Omid; Durocher, Jean; Keefe, Greg

    2016-01-01

    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. PMID:27429469

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

    PubMed

    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.

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

    PubMed

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

    PubMed Central

    Wang, Yuanyuan; Lin, Lianjie; Yin, Chunming; Othtani, Satoru; Aoyama, Katsuhiko; Lu, Changlong; Sun, Xun; Yoshikai, Yasunobu

    2014-01-01

    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. PMID:24686517

  11. Rapid residue analysis of fluoroquinolones in raw bovine milk by online solid phase extraction followed by liquid chromatography coupled to tandem mass spectrometry.

    PubMed

    Kantiani, Lina; Farré, Marinella; Barceló, Damià

    2011-12-16

    Online solid phase extraction followed by liquid chromatography coupled to tandem mass spectrometry was used for the determination of 15 fluoroquinolones and one quinolone antibiotic in raw bovine and skimmed commercial milk. Milk samples were partially deproteinized and defatted by 15 min centrifugation and then subjected to online solid phase extraction. Chromatographic separation was achieved in less than 15 min. Identification and quantification of the compounds of interest were performed by selected reaction monitoring, using an electrospray ionization source. Relative recoveries were calculated for raw milk (at 0.5, 1 and 1.5 times the MRL, or assuming 30 ng/mL for non permitted compounds) and skimmed milk (at 30 ng/mL) and ranged between 65% and 123% (raw milk) or 78% and 109% (skimmed milk). Matrix interferences were also assessed and yielded significant suppression and enhancement effects, which, at the time of quantification, were corrected by the use of six isotopically labeled standards. Quality parameters of the method were established, and limits of detection of selected antibiotics in raw milk ranged from 0.01 to 1.93 ng/mL, while in skimmed milk from 0.03 to 4.23 ng/mL. The validation of the method has been carried out according to the requirements set by the 2002/657/EC regulation. Finally, the applicability of the method was tested by analysing 28 milk samples.

  12. Effect of lactation number, year, and season of initiation of lactation on milk yield of cows hormonally induced into lactation and treated with recombinant bovine somatotropin.

    PubMed

    Mellado, M; Antonio-Chirino, E; Meza-Herrera, C; Veliz, F G; Arevalo, J R; Mellado, J; de Santiago, A

    2011-09-01

    Records representing data from 1,500 barren Holstein cows over an 8-yr period from a large commercial dairy farm in northern Mexico were analyzed to determine the effects of lactation number and season and year of initiation of lactation on milk production of cows induced hormonally into lactation and treated with recombinant bovine somatotropin (rbST) throughout lactation. Peak and 305-d milk yields were also assessed as predictors of total milk yield in cows induced into lactation. A significant quadratic relationship was found between 305-d milk yield and number of lactation [7,607±145 and 9,548±181 kg for first- and ≥6-lactation cows, respectively; mean ± standard error of the mean (SEM)] with the highest production occurring in the fifth lactation. Total milk yields of cows with ≤2 lactations were approximately 4,500 kg less than milk yields of adult cows (the overall average ± standard milk yield was 13,544±5,491 kg per lactation and the average lactation length was 454±154 d). Moreover, 305-d milk production was depressed in cows induced into lactation in spring (8,804±153 kg; mean ± SEM) and summer (8,724±163 kg) than in fall (9,079±151 kg) and winter (9,085±143 kg). Partial regression coefficients for 305-d milk yield and peak milk yield indicated an increment of 157 kg of milk per lactation per 1-kg increase in peak milk yield (r(2)=0.69). Neither peak milk yield (r(2)=0.18) nor 305-d milk yield (r(2)=0.29) was accurate for predicting total milk yield per lactation. Year, parity, and season effects had significant influence on milk yield of cows induced into lactation and treated with rbST throughout lactation, and peak milk yield can assist in the prediction of 305-d milk yield but not total milk yield. This study also showed that hormonal induction of lactation in barren high-yielding cows is a reliable, practical, and affordable technique in countries where rbST treatment and prolonged steroid administration of dairy cows are legally

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

    PubMed

    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.

  14. Prevalence, Virulence Potential, and Antibiotic Susceptibility Profile of Listeria monocytogenes Isolated From Bovine Raw Milk Samples Obtained From Rajasthan, India.

    PubMed

    Sharma, Sanjita; Sharma, Vishnu; Dahiya, Dinesh Kumar; Khan, Aarif; Mathur, Manisha; Sharma, Amit

    2017-03-01

    Listeriosis is a serious foodborne disease of a global concern, and can effectively be controlled by a continuous surveillance of the virulent and multidrug-resistant strains of Listeria monocytogenes. This study was planned to investigate prevalence of L. monocytogenes in bovine raw milk samples. A total of 457 raw milk samples collected from 15 major cities in Rajasthan, India, were analyzed for the presence of L. monocytogenes by using standard microbiological and molecular methods. Five of the 457 samples screen tested positive for L. monocytogenes. Multiplex serotyping showed that 3/5 strains belonged to serotype 4b followed by one strain each to 1/2a and to 1/2c. Further virulence potential assessment indicated that all strains possessed inlA and inlC internalins, and, in addition, two strains also possessed the gene for inlB. All strains were positive for Listeriolysin O (LLO) and showed phosphatidylinositol-specific phospholipase C (PI-PLC) activity on an in vitro agar medium with variations in production levels among the strains. A good correlation between the in vitro pathogenicity test and the chick embryo test was observed, as the strains showing higher LLO and PI-PLC activity were found to be lethal to fertilized chick embryos. All strains were resistant to the majority of antibiotics and were designated as multidrug-resistant strains. However, these strains were susceptible to 9 of the 22 tested antibiotics. The maximum zone of inhibition (mm) and acceptable minimum inhibitory concentration were observed with azithromycin, and thus it could be the first choice of a treatment. Overall, the presence of multidrug-resistant L. monocytogenes strains in the raw milk of Rajasthan region is an indicator of public health hazard and highlighting the need of consumer awareness in place and implementation of stricter food safety regulations at all levels of milk production.

  15. Effect of bovine somatotropin and rumen-undegradable protein on mammary growth of prepubertal dairy heifers and subsequent milk production.

    PubMed

    Capuco, A V; Dahl, G E; Wood, D L; Moallem, U; Erdman, R E

    2004-11-01

    Rapid body growth during the prepubertal period may be associated with reductions in mammary parenchymal growth and subsequent milk yield. The objective of this study was to test effects of dietary rumen-undegradable protein (RUP) and administration of recombinant bovine somatotropin (bST) during the prepubertal period on mammary growth and milk yield of dairy heifers. Seventy-two Holstein heifers were used in the experiment. At 90 d of age, 8 heifers were slaughtered before initiation of treatment. Remaining heifers were assigned randomly to 1 of 4 treatments. Treatments consisted of a control diet (5.9% RUP, 14.9% CP, DM basis) or RUP-supplemented diet (control diet plus 2% added RUP) with or without 0.1 mg of bST/kg of BW per day applied in a 2 x 2 factorial design. A total of 6 heifers per treatment (3 each at 5 and 10 mo of age) were slaughtered for mammary tissue analysis. Remaining heifers were bred to evaluate impact of treatment on subsequent milk yield and composition. Mammary parenchymal growth was not affected by RUP or bST treatment. Total parenchymal mass increased from 16 to 364 g, and parenchymal DNA from 58 to 1022 mg from 3 to 10 mo of age, respectively. Furthermore, number of mammary epithelial cells likely was not affected by diet or bST because the epithelial cell proliferation index, assessed by Ki-67 labeling, was not affected by treatment, nor was total parenchymal DNA and lipid content. Neither deleterious effects of increased rates of gain nor positive effects of bST were evident in prepubertal mammary growth. Subsequent milk production and composition was not different among treatments.

  16. Sensory analysis and species-specific PCR detect bovine milk adulteration of frescal (fresh) goat cheese.

    PubMed

    Golinelli, L P; Carvalho, A C; Casaes, R S; Lopes, C S C; Deliza, R; Paschoalin, V M F; Silva, J T

    2014-11-01

    The Brazilian market for dairy products made from goat milk is increasing despite the seasonality of production and naturally small milk production per animal, factors that result in high-priced products and encourage fraud. In Brazil, no official analytical method exists for detecting adulteration of goat dairy products with cow milk. The aim of this study was to design a strategy to investigate the adulteration of frescal (fresh) goat cheeses available in the Rio de Janeiro retail market, combining analysis of cheese composition and the perception of adulteration by consumers. Commercial goat cheeses were tested by using a duplex PCR assay previously designed to authenticate cheeses, by targeting the mitochondrial 12S ribosomal RNA genes of both species simultaneously. The PCR test was able to detect 0.5% (vol/vol) cow milk added during goat cheese formulation. The analysis of 20 locally produced goat cheeses (20 lots of 4 brands) showed that all were adulterated with cow milk, even though the labels did not indicate the addition of cow milk. To estimate the ability of consumers to perceive the fraudulent addition of cow milk, a triangle test was performed, in which cheeses formulated with several different proportions of goat and cow milk were offered to 102 regular consumers of cheese. Detection threshold analysis indicated that almost half of the consumers were able to perceive adulteration at 10% (vol/vol) cow milk. Effective actions must be implemented to regulate the market for goat dairy products in Brazil, considering the rights and choices of consumers with respect to their particular requirements for diet and health, preference, and cost.

  17. Development of an analytical method for cephapirin and its metabolite in bovine milk and serum by liquid chromatography with UV-VIS detection and confirmation by thermospray mass spectometry.

    PubMed

    Tyczkowska, K L; Voyksner, R D; Aronson, A L

    1991-03-01

    Metabolites of the cephapirin beta-lactam antibiotic have not previously been reported in bovine milk. The principal metabolite was tentatively identified as desacetylcephapirin by liquid chromatography with UV-VIS photodiode array (LC/UV-VIS PDA), and liquid-chromatography-mass-spectrometric (LC-MS) detection. Synthetic desacetylcephapirin was prepared by incubation of cephapirin in bovine milk and serum at 37 degrees C. Also, a method for determining cephapirin in bovine milk and serum was developed. The detection limits for cephapirin and desacetylcephapirin were estimated to be 10 and 50 micrograms/kg, respectively, for LC/UV-VIS PDA, and 100 and 500 micrograms/kg for LC-MS.

  18. Bovine leukemia virus infection in cattle of China: Association with reduced milk production and increased somatic cell score.

    PubMed

    Yang, Y; Fan, W; Mao, Y; Yang, Z; Lu, G; Zhang, R; Zhang, H; Szeto, C; Wang, C

    2016-05-01

    The main objective of this study was to investigate the individual cow effect of bovine leukemia virus (BLV) infection on milk production and somatic cell score (SCS). The fluorescence resonance energy transfer (FRET) quantitative PCR established in this study and a commercial ELISA kit revealed that 49.1% of dairy cattle (964/1,963) from 6 provinces of China and 1.6% of beef cattle (22/1,390) from 15 provinces were BLV positive. In a detailed study of 105 cows, BLV was found most commonly in buffy coat samples that also had highest copy numbers (10(4.75±1.56) per mL); all cows negative for BLV in buffy coat samples were also negative in vaginal swab, milk, and fecal samples. Copy numbers of BLV were 10(2.90±0.42)/gram of feces, 10(0.83±0.62)/mL of milk, and 10(2.18±0.81) per vaginal swab. The BLV-positive cows had significantly lower milk production in the early (26.8 vs. 30.9kg) and middle stages of lactation (22.2 vs. 26.1kg) in animals with ≥4 parities than the BLV-negative cows; they also had significantly higher SCS in early and middle lactation stages (early=5.2 vs. 4.3; middle=4.9 vs. 3.9) in animals with ≥4 parities. Milk production and SCS did not significantly differ between the BLV-infected and -uninfected cows when they were in the late lactation stage or in animals with ≤3 parities. Taken together, our results indicate that BLV infections are widespread in the dairy farms of China. Vaginal secretions and feces may be involved in BLV transmission. A BLV infection may result in reduced milk yield and increased SCS in a parity and lactation stage-restricted manner. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

    PubMed

    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

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

    PubMed Central

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

    2010-01-01

    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

  1. Bovine milk fatty acid profiles produced by feeding diets containing solin, flax and canola.

    PubMed

    Ward, A T; Wittenberg, K M; Przybylski, R

    2002-05-01

    Fatty acid profiles with emphasis on linoleic, linolenic, oleic, and conjugated linoleic acid (CLA) were compared in milk from dairy cows fed diets containing 3.25% supplemental fat and a control diet containing no supplemented fat. The fat was supplied by either whole ground solin, flax, or canola oilseed. Solin (linola) is a new cultivar of flax that contains 28% linoleic acid in the seed, Twelve multiparous Holstein cows were assigned to one of four dietary treatments. The experimental design was a 4 x 4 Latin square with each period consisting of 16 d for adjustment to the diet followed by a 5-d sampling period. Feed intake, milk yield, milk fat yield, and milk fat percentage were not affected by treatment. Adding solin, flax, or canola oilseed to lactation diets produced the highest proportions of linoleic (C18:2), linolenic (C18:3), and oleic (C18:1) acids, respectively, in the lipid fraction of the milk of the cows consuming these diets. The proportions of C6:0 to C16:1 were depressed in the milk fat of cows fed the oilseed diets, compared with the control diet. Increasing the lactation diet levels of C18:2, by using different oilseeds, increased CLA to 1.5% of milk fatty acids. The content of CLA in milk fatty acids, however, did not increase with the solin-supplemented diet compared with the canola-supplemented diet even though the C18:2 level was higher in the former diet.

  2. Virulence factors and ability of staphylococci from bovine milk and the cowshed environment to biofilm formation.

    PubMed

    Kot, B; Binek, T; Piechota, M; Wolska, K M; Zdunek, E; Platkowska, K

    2013-01-01

    The aim of this study was to examine virulence factors and the ability of S. aureus and CNS species isolated from milk of cows with mastitis to form biofilm, and to compare them with virulence factors of staphylococci from milk of cows without mastitis and cowshed environment. Most of S. aureus strains from cows with mastitis showed haemolytic activity (93.9%), among them 72.7% and 21.2% produced alpha- and beta-haemolysin, respectively. S. aureus from cows with mastitis symptoms produced proteases (above 48%) and esterase (42.4%). The highly significant relationship between the number of S. xylosus strains producing haemolysins (62%) and the origin of these strains from milk of cows with mastitis was observed. The ability to produce proteases was significantly associated with S. sciuri from milk of cows with mastitis. The ability of biofilm formation by staphylococcal strains from milk of cows with mastitis was greater than in strains from milk of cows without mastitis and the difference was significant (p < or = 0.05). The highest percentage of strains from milk of cows with mastitis were weak biofilm formers (48.6%), while 40% and 11.4% of strains were moderate and strong biofilm producers, respectively. S. xylosus showed the highest ability to form biofilm, while the lowest ability to form biofilm was observed in S. aureus and S. epidermidis. In conclusion, production of exotoxins and enzymes, and ability of biofilm formation shown by many CNS isolated from milk of cows with mastitis symptoms indicates that these features are important in pathogenesis of this disease.

  3. In vivo digestion of bovine milk fat globules: effect of processing and interfacial structural changes. I. Gastric digestion.

    PubMed

    Gallier, Sophie; Cui, Jack; Olson, Trent D; Rutherfurd, Shane M; Ye, Aiqian; Moughan, Paul J; Singh, Harjinder

    2013-12-01

    The aim was to study the in vivo gastric digestion of fat globules in bovine cream from raw, pasteurised or pasteurised and homogenised milk. Fasted rats were gavaged once and chyme samples were collected after 30, 120 and 180 min post-gavage. Proteins from raw (RC) and pasteurised (PC) creams appeared to be digested faster and to a greater extent. Free fatty acids (FAs) increased throughout the 3h postprandial period. Short and medium chain FAs were released more rapidly than long chain FAs which were hydrolysed to a greater degree from PC. The size of the fat globules of all creams increased in the stomach. Protein aggregates were observed in pasteurised and homogenised cream chyme. Protrusions, probably caused by the accumulation of insoluble lipolytic products, appeared at the surface of the globules in RC and PC chyme. Overall, PC proteins and lipids appeared to be digested to a greater extent. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Experimental colonization of the bovine teat duct with Corynebacterium bovis and the effect on milk somatic cell counts.

    PubMed Central

    Brooks, B W; Barnum, D A

    1984-01-01

    Colonization with Corynebacterium bovis was established in 59 of 64 (92%), 58 of 59 (98%) and 19 of 34 (56%) of uninfected bovine mammary quarters following inoculation of 83.3 X 10(4) colony-forming units (CFU) of the organism into the teat cistern, 4.7 X 10(3) CFU 5 mm into the teat duct or by exposure of the teat orifice to a milk culture containing 1.6 X 10(7) CFU/mL respectively. Mean somatic cell counts for foremilk samples from 122 quarters were significantly higher after colonization with C. bovis (145,900/mL) compared to before exposure (130,900/mL). PMID:6722643

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

    PubMed

    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.

  6. Genetic variability of lactoferrin content estimated by mid-infrared spectrometry in bovine milk.

    PubMed

    Soyeurt, H; Colinet, F G; Arnould, V M-R; Dardenne, P; Bertozzi, C; Renaville, R; Portetelle, D; Gengler, N

    2007-09-01

    The effects of lactoferrin (LF) on the immune system have already been shown by many studies. Unfortunately, the current methods used to measure LF levels in milk do not permit the study of the genetic variability of lactoferrin or the performance of routine genetic evaluations. The first aim of this research was to derive a calibration equation permitting the prediction of LF in milk by mid-infrared spectrometry (MIR). The calibration with partial least squares on 69 samples showed a ratio of standard error of cross-validation to standard deviation equal to 1.98. Based on this value, the calibration equation was used to establish an LF indicator trait (predicted LF; pLF) on a large number of milk samples (n = 7,690). A subsequent study of its variability was conducted, which confirmed that stage of lactation and lactation number influence the overall pLF level. Small differences in mean pLF among 7 dairy breeds were also observed. The pLF content of Jersey milk was significantly higher than that in Holstein milk. Therefore, the choice of breed could change the expected LF level. Heritability estimated for pLF was 19.7%. The genetic and phenotypic correlations between somatic cell score and pLF were 0.04 and 0.26, respectively. As somatic cell score increases in presence of mastitis, this observation seems to indicate that pLF, or a function of observed pLF, compared with expected LF might have potential as an indicator of mastitis. The negative genetic correlation (-0.36) between milk yield and pLF could indicate an undesirable effect of selection for high milk production on the overall LF level.

  7. Genetic relatedness and virulence factors of bovine Staphylococcus aureus isolated from teat skin and milk.

    PubMed

    da Costa, L B; Rajala-Schultz, P J; Hoet, A; Seo, K S; Fogt, K; Moon, B S

    2014-11-01

    The objective of this study was to assess the role of teat skin colonization in Staphylococcus aureus intramammary infections (IMI) by evaluating genetic relatedness of Staph. aureus isolates from milk and teat skin of dairy cows using pulsed-field gel electrophoresis and characterizing the isolates based on the carriage of virulence genes. Cows in 4 known Staph. aureus-positive herds were sampled and Staph. aureus was detected in 43 quarters of 20 cows, with 10 quarters positive in both milk and skin (20 isolates), 18 positive only in milk, and 15 only on teat skin. Quarters with teat skin colonized with Staph. aureus were 4.5 times more likely to be diagnosed with Staph. aureus IMI than quarters not colonized on teat skin. Three main clusters were identified by pulsed-field gel electrophoresis using a cutoff of 80% similarity. All 3 clusters included both milk and skin isolates. The majority of isolates (72%) belonged to one predominant cluster (B), with 60% of isolates in the cluster originating from milk and 40% from teat skin. Genotypic variability was observed within 10 pairs (formed by isolates originating from milk and teat skin of the same quarter), where isolates in 5 out of the 10 pairs belonged to the same cluster. Forty-two virulence factors were screened using PCR. Some virulence factors were carried more frequently by teat skin isolates than by milk isolates or isolates from quarters with high somatic cell counts. Isolates in the predominant cluster B carried virulence factors clfA and clfB significantly more often than isolates in the minor clusters, which may have assisted them in becoming predominant in the herds. The present findings suggest that teat skin colonization with Staph. aureus can be an important factor involved in Staph. aureus IMI.

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

    PubMed

    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.

  9. Bovine serum albumin and insulin-dependent diabetes mellitus; is cow's milk still a possible toxicological causative agent of diabetes?

    PubMed

    Persaud, Deryck R; Barranco-Mendoza, Alma

    2004-05-01

    The implication of bovine serum albumin (BSA) in cow's milk as a causative agent for the onset of insulin-dependent diabetes mellitus (IDDM) is a major topic of scientific debate not withstanding the medical and economic implications. A critical survey of the pertinent literature has revealed a number of controversies. For example, an important toxicological aspect of BSA is the presence of ABBOS, a peptide segment of the protein. However, the nature and effect of ABBOS on the death of insulin producing cells (beta-cells of the pancreas) is unclear and hence inconclusive. In addition, studies in diabetes-prone mice and rats appear to show that cow's milk does not alter the frequency of diabetes in these organisms. It is suggested that BSA may not be the cause of diabetes. Instead, IDDM is most likely the result of oxidative stress, due to high local levels of nitric oxide (NO*) and oxygen radicals (O2*-), on the beta-cells of the pancreas, which eventually leads to their destruction.

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

    PubMed

    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.

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

    PubMed Central

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

    2015-01-01

    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. PMID:26074238

  12. Detection of Helicobacter pylori glmM gene in bovine milk using Nested polymerase chain reaction

    PubMed Central

    Osman, Eyman Y.; El-Eragi, A. M. S.; Musa, Abuobeida M.; El-Magboul, Salma B.; A/Rahman, Magdi B.; Abdo, Abdelmounem E.

    2015-01-01

    Aim: The aim was to detect the glmM gene of Helicobacter pylori (H. pylori) in cow’s milk from different dairy farms in Khartoum State using Nested polymerase chain reaction (PCR). Materials and Methods: A total of 50 milk samples were collected from different dairy farms in Khartoum State (13 from Khartoum, 24 Khartoum North, and 13 from Omdurman Provinces). Results: The generated results showed that 11/50 (22%) were harboring the investigated H. pylori glmM gene in Khartoum State (1/13 [7.7%] Khartoum, 9/24 [37.5%] Khartoum North, and 1/13 [7.7%] Omdurman provinces, respectively). Conclusion: To the best of our knowledge, this was the first report on the detection of H. pylori glmM gene in cattle milk in Khartoum State. Nonetheless, the high percentages of H. pylori DNA detection in milk opened new avenues toward exploring the risk of human infection with H. pylori through the consumption of raw milk. PMID:27047175

  13. Evaluation of a novel chemical sensor system to detect clinical mastitis in bovine milk.

    PubMed

    Mottram, Toby; Rudnitskaya, Alisa; Legin, Andrey; Fitzpatrick, Julie L; Eckersall, P David

    2007-05-15

    Automatic detection of clinical mastitis is an essential part of high performance and robotic milking. Currently available technology (conductivity monitoring) is unable to achieve acceptable specificity or sensitivity of detection of clinical mastitis or other clinical diseases. Arrays of sensors with high cross-sensitivity have been successfully applied for recognition and quantitative analysis of other multicomponent liquids. An experiment was conducted to determine whether a multisensor system ("electronic tongue") based on an array of chemical sensors and suitable data processing could be used to discriminate between milk secretions from infected and healthy glands. Measurements were made with a multisensor system of milk samples from two different farms in two experiments. A total of 67 samples of milk from both mastitic and healthy glands were in two sets. It was demonstrated that the multisensor system could distinguish between control and clinically mastitic milk samples (p=0.05). The sensitivity and specificity of the sensor system (93 and 96% correspondingly) showed an improvement over conductivity (56 and 82% correspondingly). The multisensor system offers a novel method of improving mastitis detection.

  14. Bacteriophage Types and Antibiotic Sensitivity of Staphylococci from Bovine Milk and Human Nares1

    PubMed Central

    Jones, R. H.; Bennett, F. W.

    1965-01-01

    The number, phage types, and antibiotic sensitivity of coagulase-positive staphylococci from grade A raw milk samples produced on 40 farms in the Athens, Ga., milkshed were determined. Counts of mannitol-positive staphylococci in milk ranged from 100 to 3,580 per milliliter, with an arithmetic mean of 1,047. Examination of the nares of 48 dairymen on 34 of the farms also revealed that 13 (27%) were carriers of coagulase-positive staphylococci. Isolates from milk (412) and from nares (39) were tested against the Coles, Seto-Wilson, and International phage sets and 87, 68, and 56%, respectively, proved typable. Nine isolates were not typable. Each of the 33 phages used lysed one or more of the isolates. Staphylococcal phage types per milk sample ranged from 0 to 5, 0 to 7, and 0 to 8, with arithmetic means of 1.9, 2.3, and 2.3, respectively. Of the 13 narial carriers, 7 harbored staphylococci of one or more of the same phage types as those isolated from the milk at the respective farms. Randomly selected isolates were tested against high and low concentrations of 12 common antibiotics. All were either moderately sensitive or resistant to polymixin B. Over 30% were moderately sensitive or resistant to dihydrostreptomycin and penicillin individually. With but few exceptions, all isolates were sensitive to chlortetracycline, bacitracin, carbomycin, chloramphenicol, erythromycin, neomycin, novobiocin, oxytetracycline, and tetracycline individually. PMID:5325936

  15. Evaluation of biofilm formation using milk in a flow cell model and microarray characterization of Staphylococcus aureus strains from bovine mastitis.

    PubMed

    Snel, G G M; Malvisi, M; Pilla, R; Piccinini, R

    2014-12-05

    It was hypothesized that biofilm could play an important role in the establishment of chronic Staphylococcus aureus bovine mastitis. The in vitro evaluation of biofilm formation can be performed either in closed/static or in flow-based systems. Efforts have been made to characterize the biofilm-forming ability of S. aureus mastitis isolates, however most authors used static systems and matrices other than UHT milk. It is not clear whether such results could be extrapolated to the mammary gland environment. Therefore, the present study aimed to investigate the biofilm-forming ability of S. aureus strains from subclinical bovine mastitis using the static method and a flow-based one. One hundred and twelve strains were tested by the classic tissue culture plate assay (TCP) and 30 out of them were also tested by a dynamic semi-quantitative assay using commercial UHT milk as culture medium (Milk Flow Culture, MFC) or Tryptic Soy Broth as control medium (TS Flow Culture, TSFC). Only 6 (20%) strains formed biofilm in milk under flow conditions, while 36.6% were considered biofilm-producers in TCP, and 93.3% produced biofilm in TSFC. No agreement was found between TCP, MFC and TSFC results. The association between strain genetic profile, determined by microarray, and biofilm-forming ability in milk was evaluated. Biofilm formation in MFC was significantly associated with the presence of those genes commonly found in bovine-associated strains, assigned to clonal complexes typically detected in mastitis. Based on our results, biofilm-forming potential of bovine strains should be critically analysed and tested applying conditions similar to mammary environment. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Bacterial Community Profiling of Milk Samples as a Means to Understand Culture-Negative Bovine Clinical Mastitis

    PubMed Central

    Kuehn, Joanna S.; Gorden, Patrick J.; Munro, Daniel; Rong, Ruichen; Dong, Qunfeng; Plummer, Paul J.; Wang, Chong; Phillips, Gregory J.

    2013-01-01

    Inflammation and infection of bovine mammary glands, commonly known as mastitis, imposes significant losses each year in the dairy industry worldwide. While several different bacterial species have been identified as causative agents of mastitis, many clinical mastitis cases remain culture negative, even after enrichment for bacterial growth. To understand the basis for this increasingly common phenomenon, the composition of bacterial communities from milk samples was analyzed using culture independent pyrosequencing of amplicons of 16S ribosomal RNA genes (16S rDNA). Comparisons were made of the microbial community composition of culture negative milk samples from mastitic quarters with that of non-mastitic quarters from the same animals. Genomic DNA from culture-negative clinical and healthy quarter sample pairs was isolated, and amplicon libraries were prepared using indexed primers specific to the V1–V2 region of bacterial 16S rRNA genes and sequenced using the Roche 454 GS FLX with titanium chemistry. Evaluation of the taxonomic composition of these samples revealed significant differences in the microbiota in milk from mastitic and healthy quarters. Statistical analysis identified seven bacterial genera that may be mainly responsible for the observed microbial community differences between mastitic and healthy quarters. Collectively, these results provide evidence that cases of culture negative mastitis can be associated with bacterial species that may be present below culture detection thresholds used here. The application of culture-independent bacterial community profiling represents a powerful approach to understand long-standing questions in animal health and disease. PMID:23634219

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

    PubMed

    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

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

    PubMed

    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.

  19. Comparison between mechanical and near-infrared methods for assessing coagulation properties of bovine milk.

    PubMed

    Cipolat-Gotet, C; Cecchinato, A; De Marchi, M; Penasa, M; Bittante, G

    2012-11-01

    The aim of the present study was to compare milk coagulation properties measured through a traditional mechanical device, the Formagraph (FRM; Foss Electric A/S, Hillerød, Denmark), and a near-infrared optical device, the Optigraph (OPT; Ysebaert SA, Frépillon, France). Individual milk samples of 913 Brown Swiss cows from 63 herds located in Trento Province (Italy) were analyzed for rennet coagulation time (RCT, min), curd-firming time (k(20), min), and 2 measures of curd firmness (a(30) and a(45),mm) using the 2 instruments and under identical conditions. The trial was performed in the same laboratory, by the same technician, and following the same procedures. Extending the analysis by either instrument to 90 min permitted RCT and k(20) values to be obtained even for late-coagulating milk samples. Milk coagulation properties measured using the OPT differed considerably from those obtained using the FRM. The average k(20) values varied greatly (8.16 vs. 5.36 min for the OPT and the FRM, respectively), as did the a(45) figures (41.49 vs. 33.66 mm for the OPT and the FRM, respectively). The proportion of noncoagulating samples for which k(20) could be estimated differed between instruments, being less for the OPT. The between-instrument correlation coefficients were either moderate (0.48 for a(30)) or low (0.24 and 0.17 for k(20) and a(45), respectively) when the same traits were compared. The correlations between k(20) and a(45), and milk yield varied among instruments, as did the correlations between k(20), a(30), and a(45) and milk composition, and the correlations between a(45) and pH. The relative influence of days in milk on k(20) and a(45) varied, as did the effect of parity on a(45) and that of the measuring unit of coagulation meter on k(20) and a(30). The RCT estimated by the OPT was the only milk coagulation property to show good agreement with the FRM-derived value, although this was not true for the data from late-coagulating samples.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  1. Antimicrobial resistance and genotypes of staphylococci from bovine milk and the cowshed environment.

    PubMed

    Kot, B; Piechota, M; Antos-Bielska, M; Zdunek, E; Wolska, K M; Binek, T; Olszewska, J; Guliński, P; Trafny, E A

    2012-01-01

    Investigation of antimicrobial resistance and genetic relatedness of staphylococci from milk of cows with mastitis and cowshed environment was the aim of this study. Antimicrobial resistance against 14 antimicrobials were determined by using a disc diffusion method. Genetic similarity between the most frequently isolated species was analysed by PFGE (pulsed-field gel electrophoresis). Haemolytic activity, DNase, protease and esterase production was also investigated. Coagulase-negative Staphylococcus species were isolated from 30.8% of milk samples from cows with mastitis. The most frequently isolated species was Staphylococcus xylosus and yield of these organisms was significantly associated with milk of mastitis cows. S. epidermidis was a predominant penicillin-resistant species. High frequency of resistance to lincomycin was observed among isolates of S. sciuri (54.2%) and S. xylosus (25.9%) from cows with mastitis. PFGE (pulsed-field gel electrophoresis) analysis of 29 Staphylococcus aureus isolates showed the presence of 17 PFGE pulsotypes. Isolates of S. sciuri (n = 36) had unique PFGE patterns. Some S. xylosus isolates from milk and milker's hands had the same PFGE pulsotypes, and this observation could indicate that dairyman may be a potential source of the infection. The pulsotype of each of the remaining isolates of S. xylosus suggested that they might have come from common environmental sources; however, these isolates differed in antibiotic resistance pattern or virulence traits. Therefore, knowledge about antibiotic sensitivity pattern and virulence factors of a CNS isolate, besides its genotype, may be informative in tracking the source of the infection.

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

    PubMed

    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.

  3. Xanthine dehydrogenase to xanthine oxidase conversion in ischemic rat intestine

    SciTech Connect

    McKelvey, T.G.; Engerson, T.D.; Elmore, C.R.; Jones, H.P. )

    1990-02-26

    The ischemic conversion of the NADH-producing xanthine dehydrogenase (XDH) to an oxidase form, that produces both superoxide radical and hydrogen peroxide, has been proposed as an important step in initiating oxygen radical-mediated ischemia-reperfusion injury. It has also been reported that two forms of converted oxidase are produced in ischemic rat liver; a reversible xanthine oxidase produced through sulfhydryl oxidation, that can be reconverted to XDH by incubation with 10mM dithiothreitol (Dtt) at 37{degrees}C, and a Dtt-irreversible oxidase produced via proteolysis. The authors report that increased oxidase in the ischemic rat intestine results from significant increases in both the Dtt-reversible and Dtt-irreversible forms of xanthine oxidase. Total oxidase activity (Irreversible + Dtt-reversible) was 19% of the total enzyme activity (XDH + XO) in control ileum and distal jejunum, increased to 26% after 1 hour of ischemia at 37{degrees}C, and significantly to 36% after 1.5 hours. After 3 hours 73% of the activity was in the oxidase form. Irreversible oxidase comprised 15% of the total activity in control intestine, significantly increased to 25% after 2 hours, and further to 42% after 3 hours. Dtt-reversible oxidase was 3% of the total activity in controls, increased to 13% after 1.5 hours, and significantly to 29% after 2 hours.

  4. Biovars of coagulase-positive staphylococci isolated from bovine mastitic milk.

    PubMed

    García López, M L; García Fernández, M C; García Armesto, M R; Prieto, M; Otero, A; Fernández Alvarez, M F

    1990-06-01

    Fifty strains of coagulase-positive staphylococci associated with bovine mastitis were biotyped. Of them, 20 were identified as biovar C (cattle & sheep), 17 as biovar B (poultry & swine), 2 as biovar D (hares) and 1 as biovar F (pigeon & fox). Of the remaining strains, 2 were closely related to human biovar A, 2 failed one property to be classified as biovar B, 3 shared properties of biovars B and D and 3 could not be identified. Bovine strains belonging to biovar C formed an heterogeneous group showing differences in crystal violet growth type and production of alpha haemolysin. Only strains associated with biovar A were Tween 80 positive. The highest incidence of lytic reactions amongst all biovars was with phages 42 E (III) and 102 (IV). Antibiotic resistance was most frequently found in biovar C (65%). The 2 strains in biovar D, one classed as biovar C and one as intermediate, were enterotoxigenic (C or D toxins).

  5. Organ distribution and molecular forms of human xanthine dehydrogenase/xanthine oxidase protein.

    PubMed

    Sarnesto, A; Linder, N; Raivio, K O

    1996-01-01

    Xanthine dehydrogenase/xanthine oxidase (XDH/XO) is a major cytoplasmic source of superoxide radicals and hydrogen peroxide, and it is considered important in the pathogenesis of ischemia-reperfusion damage. Because little is known about the enzyme in human tissues, the aims of this study were to purify human XDH/XO and to produce Ab for detection of the protein in Western blots and for quantification by ELISA. We purified human milk XDH/XO, produced Ab for Western blotting and ELISA of the protein, and evaluated the molecular forms and activity-protein relationships in human tissues. The molecular size of the purified protein under nondenaturing conditions was approximately 300 kd. On SDS-PAGE, it was fragmented into four main bands of 143, 125, 87, and 59 kd. Ab recognized bands of similar size in Western blots of the purified preparation and human milk. In fresh liver homogenates treated with anti-proteases, the three largest bands were observed; in the intestine, only the two largest were observed. Serum, brain, heart, and skeletal muscle were negative, whereas some lung and kidney samples showed one faint band of 143 kd. Trypsin treatment of the enzyme converted the large molecular-weight bands into smaller bands, as did incubation of a liver homogenate without anti-proteases. XDH/XO protein concentrations (ng/mg total protein) were 146 +/- 70 in liver and 556 +/- 320 in intestine and less than 5 ng/ml in serum. The relationship of activity to protein (2.7-3.0 mumol/min/mg XDH/XO protein) was constant in liver and intestine during development. We conclude that 1) human XDH/XO has molecular size and subunit structure similar to other mammalian enzymes; 2) the polypeptide chain is unstable, also in the intact cell, despite retained activity; and 3) the amount of inactive XDH/XO in human liver and intestine is apparently small.

  6. 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.

    PubMed

    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.

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

    SciTech Connect

    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.

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

    PubMed

    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

  9. Xanthine oxidase inhibition attenuates ischemic-reperfusion lung injury

    SciTech Connect

    Lynch, M.J.; Grum, C.M.; Gallagher, K.P.; Bolling, S.F.; Deeb, G.M.; Morganroth, M.L.

    1988-05-01

    Ischemic-reperfusion lung injury is a factor potentially limiting the usefulness of distant organ procurement for heart-lung transplantation. Toxic oxygen metabolites are considered a major etiologic factor in reperfusion injury. Although oxygen-free radicals may be generated by many mechanisms, we investigated the role of xanthine oxidase in this injury process by using lodoxamide, a xanthine oxidase inhibitor, to inhibit ischemic-reperfusion injury in an isolated rat lung model. Isolated rat lungs were perfused with physiologic salt solution (PSS) osmotically stabilized with Ficoll until circulating blood elements were nondetectable in the pulmonary venous effluent. Lungs were rendered ischemic by interrupting ventilation and perfusion for 2 hr at 37/sup 0/C. After the ischemic interval, the lungs were reperfused with whole blood and lung injury was determined by measuring the accumulation of /sup 125/I-bovine serum albumin in lung parenchyma and alveolar lavage fluid as well as by gravimetric measurements. Lung effluent was collected immediately pre- and postischemia for analysis of uric acid by high-pressure liquid chromatography. Lodoxamide (1 mM) caused significant attenuation of postischemic lung injury. Uric acid levels in the lung effluent confirmed inhibition of xanthine oxidase. Protection from injury was not complete, however, implying that additional mechanisms may contribute to ischemic-reperfusion injury in the lung.

  10. 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.

    PubMed

    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.

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

    PubMed Central

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

    2011-01-01

    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. PMID:21331157

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

    PubMed

    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.

  13. Comparison of identification systems for psychrotrophic bacteria isolated from raw bovine milk.

    PubMed

    Vithanage, Nuwan R; Yeager, Thomas R; Jadhav, Snehal R; Palombo, Enzo A; Datta, Nivedita

    2014-10-17

    Psychrotrophic bacteria in raw milk produce heat-resistant extracellular proteases, resulting in spoilage and shelf-life reduction of ultrahigh temperature treated milk and milk products. Controlling of these spoilage microbes requires rapid and reliable identification systems for screening of raw milk. This study aimed to compare commercial bacterial identification systems with a genetic method (considered as the 'gold standard' method) for the identification of heat-resistant protease producing bacteria in raw milk. Five bacterial identification systems were compared based on typability, discrimination power (i.e. Simpson's Index of Diversity), reproducibility and speed of analysis. The accuracy of 16S rRNA gene sequencing, Biolog, MALDI-TOF MS, API, and Microbact for the identification of Gram negative bacilli at the species level was 100.0%, 86.8%, 63.2%, 60.5% and 57.9%, respectively. The Gram positive bacilli were identified by 16S rRNA gene sequencing, Biolog, MALDI-TOF MS, and API with accuracies at the species level of 100.0%, 85.0%, 95.0% and 90.0%, respectively. The 16S rRNA gene sequencing and phylogenetic analysis discriminated Pseudomonas fluorescens, Pseudomonas syringae, Hafnia alvei, Bacillus cereus, Bacillus pumilus and Bacillus licheniformis to the subspecies level. The Simpson's Index of Diversity scores were 0.966, 0.711, 0.496, 0.472, and 0.140, for 16S rRNA gene sequencing, Biolog, MALDI-TOF MS, API and Microbact, respectively. Limited reference profiles in the databases of Biolog, MALDI-TOF MS, API and Microbact systems reduced their accuracy in bacterial identification, compared to 16S rRNA gene sequencing. The rapidity of each assay is in the following order; MALDI-TOF MS>16S rRNA gene sequencing>Biolog>Microbact>API. The reproducibility of the assays is in the order of 16S rRNA gene sequencing>API>Microbact>MALDI-TOF MS>Biolog. Thus, 16S rRNA gene sequencing appears to be the most reliable and robust system for the identification of dairy

  14. Selective, reliable blood and milk bio-markers for diagnosing clinical and subclinical bovine mastitis.

    PubMed

    Sadek, Kadry; Saleh, Ebeed; Ayoub, Mousa

    2017-02-01

    Mastitis is positioned as the most vital ailment in dairy cows in light of conventional cost examinations. The aim of the present study was to evaluate the diagnostic value of different acute phase proteins (APPs), pro-inflammatory cytokines, and oxidative stress biomarkers in healthy cows and in those with clinical or subclinical mastitis and to localize APP gene expression in the milk of mastitic cows. Therefore, 20 subclinical mastitic cows with positive California Mastitis Test (CMT) results and no clinical signs of mastitis, 15 clinically mastitic cows, and 15 healthy cows with negative CMT results and somatic cell count (SCC) of <600,000 cells/mL were selected. Milk and blood samples were collected. The present findings indicate that the biochemical parameters examined were significantly (p < 0.05) increased in cows with both types of mastitis, except for total protein, albumin, and GSH levels and the TAC, which were significantly (p < 0.05) decreased, compared with values in the controls. Surprisingly, SAA and Hp gene expression were up-regulated in milk from cows with both forms of mastitis, while Fb expression was absent. The present study demonstrates that APPs, pro-inflammatory cytokines, and indicators of oxidative stress may serve as biomarkers of clinical and subclinical mastitis. Interestingly, the expression of SAA and Hp indicates the local de novo synthesis of these APPs within the mammary glands. Furthermore, the presence of SAA and Hp transcripts in milk cells derived from pathogen-free mammary glands proved their constitutive expression. However, future studies with more extensive baseline sampling are still needed to establish and validate the reference values for APPs, cytokines, and oxidative stress markers in cows.

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

    PubMed

    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.

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

    PubMed

    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.

  17. Species distribution and in vitro antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine mastitic milk.

    PubMed

    Taponen, Suvi; Nykäsenoja, Suvi; Pohjanvirta, Tarja; Pitkälä, Anna; Pyörälä, Satu

    2016-02-06

    Coagulase-negative staphylococci (CoNS) are the most common bovine mastitis causing bacteria in many countries. It is known that resistance for antimicrobials is in general more common in CoNS than in Staphylococcus aureus but little is known about the antimicrobial resistance of specific CoNS species. In this study, 400 CoNS isolates from bovine mastitic milk samples were identified to species level using ribotyping and MALDI-TOF MS, and their antimicrobial susceptibility was determined using a commercially available microdilution system. The results were interpreted according to the epidemiological cut-off values by the European Committee on Antimicrobial Susceptibility testing. The most common CoNS species were S. simulans, S. epidermidis, S. chromogenes and S. haemolyticus. Penicillin resistance was the most common type of antimicrobial resistance. Staphylococcus epidermidis was the most resistant among the four major species. Almost one-third of our S. epidermidis isolates were resistant to >2 antimicrobials and close to 7 % were multidrug resistant. The majority of S. epidermidis isolates were resistant to benzylpenicillin. On the contrary, only few S. simulans isolates were penicillin-resistant. Phenotypic oxacillin resistance was found in all four main species, and 34 % of the isolates were oxacillin resistant. However, only 21 isolates (5 %) were positive for the mecA gene. Of these, 20 were S. epidermidis and one S. sciuri. mecC positive isolates were not found. Staphylococcus epidermidis differed from the three other major CoNS species as resistance to the tested antimicrobials was common, several isolates were multidrug resistant, and 19 % of the isolates carried the mecA gene encoding methicillin resistance.

  18. Conformational alterations induced by novel green 16-E2-16 gemini surfactant in xanthine oxidase: Biophysical insights from tensiometry, spectroscopy, microscopy and molecular modeling.

    PubMed

    Akram, Mohd; Bhat, Imtiyaz Ahmad; Bhat, Waseem Feeroze; Kabir-ud-Din

    2015-01-01

    Herein we report the interaction of a biodegradable gemini surfactant, ethane-1,2-diyl bis(N,N-dimethyl-N-hexadecylammoniumacetoxy) dichloride (16-E2-16) with bovine milk xanthine oxidase (XO), employing tensiometry, fluorescence spectroscopy, UV spectroscopy, far-UV circular dichroism spectroscopy (CD), Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and computational molecular modeling. Surface tension results depict substantial changes in the micellar as well as interfacial parameters (CMC, ΠCMC, γCMC, Γmax, Amin, ΔGmic° and ΔGads°) of 16-E2-16 gemini surfactant upon XO combination, deciphering the interaction of XO with the gemini surfactant. Fluorescence measurements reveal that 16-E2-16 gemini surfactant causes quenching in the xanthine oxidase (XO) fluorescence spectra via static procedure and the values of various evaluated binding parameters (KSV, Kb, kq, ΔGb° and n) describe that 16-E2-16 effectively binds to XO. Three dimensional fluorescence, 8-anilino-1-naphthalene sulfonic acid (ANS) binding, F1F3 ratio, UV, CD, FTIR, SEM and TEM results delineate changes in the secondary structure of xanthine oxidase. Molecular docking results provide complement to the steady-state fluorescence findings and support the view that quenching occurs due to non-polar environment experienced by aromatic residues of the enzyme. The results of this study can help scientists to tune the conformation of an enzyme (XO) with biocompatible amphiphilic microstructures, which will help to unfold further understanding in the treatment modes of various diseases like gout, hyperuricemia, liver and brain necrosis. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. 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.

    PubMed

    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.

  20. The activity of milk leukocytes in response to a water-soluble fraction of Mycobacterium phlei in bovine subclinical mastitis.

    PubMed

    Mukherjee, R; Ram, G C; Dash, P K; Goswami, T

    2004-01-01

    The effect of a water-soluble fraction (WSF) of a non-pathogenic strain of Mycobacterium phlei was studied in bovine subclinical mastitis (SCM) by measuring the myeloperoxidase and acid phosphatase enzyme levels in the milk leukocytes. Forty-five cows were divided into three equal groups. Group I, consisting of 15 healthy cows, served as the control, whereas groups II and III each contained 15 cows with subclinical mastitis on the basis of a positive reaction in the California mastitis test (CMT). The cows in group II received 100 microg of WSF in 5 ml sterile phosphate-buffered saline, pH 7.4 (PBS) once only, while those in group III received 5 ml sterile PBS daily for 7 days, both treatments being given by the intramammary route. Observations were made up to 30 days after treatment (AT). The CMT of the healthy milk was negative (0), whereas it ranged between 1 and 2 points in SCM. The somatic cell count (SCC) increased significantly (p < 0.05) on day 3, then fell steeply from day 7 up to day 30 AT in the cows in group II. A steady decrease in the total bacterial count (TBC) was observed in the group treated with WSF but the bacterial counts remained high in the groups treated with PBS. The mean acid phosphatase level was enhanced by 119% on day 3 AT in group II but only by 18.7% in the cows in group III. The mean myeloperoxidase level was enhanced by 100% in the cows in group II but only by 18% in those in group III on day 3 AT. This significant reduction in the bacterial load in infected cows caused by intramammary infusion of WSF may be due to activation of the microbicidal activity of the neutrophils, but this requires confirmation.

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

    PubMed Central

    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 <6 months of age. Objectives The aim of the present study was to characterize the oral microbiota in infants fed MFGM-supplemented formula and compare it to that of infants fed standard formula or breast milk. Methods In a prospective double-blinded randomized controlled trial, exclusively formula-fed infants <2 months of age were randomized to be fed experimental formula (EF, n = 80) with reduced energy and protein and supplemented with a bovine MFGM concentrate, or standard formula (SF, n = 80) until 6 months of age. A breast-fed reference (BFR, n = 80) group was also recruited. The oral microbiota was analyzed at 4 (n = 124) and 12 (n = 166) months of age using Illumina MiSeq multiplex sequencing and taxonomic resolution against the HOMD 16S rDNA database of oral bacteria. Results Species richness in the oral samples did not differ between the EF and SF groups, but partial least square modeling identified a few taxa that were significantly associated with being in either group, e.g. lower level of Moraxella catarrhalis in the EF group. Infants in the BFR group had significantly lower species richness at 4 months of age and their microbiota pattern differed markedly from the formula-fed groups. Conclusions Supplementation of infant formula with MFGM yielded moderate 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

  2. Evaluation of Three Bacterial Identification Systems for Species Identification of Bacteria Isolated from Bovine Mastitis and Bulk Tank Milk Samples.

    PubMed

    Savage, Emily; Chothe, Shubhada; Lintner, Valerie; Pierre, Traci; Matthews, Tammy; Kariyawasam, Subhashinie; Miller, Dawn; Tewari, Deepanker; Jayarao, Bhushan

    2017-03-01

    A study was conducted to evaluate Sensititre(®) Automated Reading and Incubation System 2x System (ARIS), API(®) (API), and Bruker MALDI-TOF MS (MALDI) bacterial species identification systems using 132 diverse bacterial isolates from bovine milk samples and bulk tank milk received at the Penn State Animal Diagnostic Laboratory. The results were compared with 16S rRNA gene sequence analysis, which served as the reference method for species identification. The ARIS, API, and MALDI identified 0%, 40%, and 33.4% of species classified as Gram-positive rod isolates belonging to genera Arthrobacter, Bacillus, Brachybacterium, Brevibacterium, and Corynebacterium, respectively. It was observed that 76.5%, 93.9%, and 96.9% of catalase-negative, Gram-positive cocci (n = 33; Aerococcus, Enterococcus, Lactococcus, Streptococcus) were correctly identified to the species level by ARIS, API, and MALDI, respectively, while 33.4%, 84.5%, and 97.7% of catalase-positive, Gram-positive cocci (n = 45; Kocuria, Staphylococcus) were correctly identified to their species by ARIS, API, and MALDI, respectively. A total of 48 isolates (Acinetobacter, Citrobacter, Enterobacter, Escherichia, Klebsiella, Pantoea, Pasteurella, Providencia, Pseduomonas, Serratia) of Gram-negative bacteria were examined, of which 85.4%, 93.7%, and 95.8% of the isolates were correctly identified to the species level by ARIS, API, and MALDI, respectively. In our laboratory, the MALDI had the least costs associated with consumables and reagents compared to ARIS, API, and 16S rRNA identification methods. Identification of bacterial species was accomplished in <2 h using MALDI and 24 h for ARIS, API, and 16S rRNA identification systems.

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

    PubMed

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

    2017-01-01

    In a recent study, supplementation of infant formula with milk fat globule membranes (MFGM) decreased the incidence of otitis media in infants <6 months of age. The aim of the present study was to characterize the oral microbiota in infants fed MFGM-supplemented formula and compare it to that of infants fed standard formula or breast milk. In a prospective double-blinded randomized controlled trial, exclusively formula-fed infants <2 months of age were randomized to be fed experimental formula (EF, n = 80) with reduced energy and protein and supplemented with a bovine MFGM concentrate, or standard formula (SF, n = 80) until 6 months of age. A breast-fed reference (BFR, n = 80) group was also recruited. The oral microbiota was analyzed at 4 (n = 124) and 12 (n = 166) months of age using Illumina MiSeq multiplex sequencing and taxonomic resolution against the HOMD 16S rDNA database of oral bacteria. Species richness in the oral samples did not differ between the EF and SF groups, but partial least square modeling identified a few taxa that were significantly associated with being in either group, e.g. lower level of Moraxella catarrhalis in the EF group. Infants in the BFR group had significantly lower species richness at 4 months of age and their microbiota pattern differed markedly from the formula-fed groups. Supplementation of infant formula with MFGM yielded moderate 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.

  4. Activity of cyclooxygenase-2 and nitric oxide in milk leucocytes following intramammary inoculation of a bio-response modifier during bovine Staphylococcus aureus subclinical mastitis.

    PubMed

    De, U K; Mukherjee, Reena

    2014-09-01

    The objective of the study was to evaluate the effects of intramammary infusion of a bio-response modifier (BRM) prepared from Nocardia globerula on certain inflammatory markers and percentage of neutrophil/lymphocyte in mammary secretions during bovine Staphylococcus aureus subclinical mastitis (SCM). The somatic cell count (SCC), total bacterial count (TBC) in milk, cyclooxygenase-2 (COX-2) activity, production of nitrite and nitrate (NOx) in milk leukocytes and neutrophil % and lymphocyte % in milk were evaluated before and after intramammary infusion of BRM in healthy and quarters inflicted with S. aureus SCM. Intramammary infusion of BRM significantly enhanced the SCC in earlier phase with subsequent reduction on day 7 after initiation of treatment. Whereas, the reduction of TBC was observed from day 3 onwards. The COX-2 activity and NOx production in milk cell increased initially on day 3 of post treatment but reduced on day 5 in SCM infected quarters following BRM infusion. The neutrophil % and lymphocyte % in milk also enhanced significantly on day 3 but reduced on day 5 in SCM infected quarters in response to BRM infusion. Initial influx of SCC, increased neutrophil%, lymphocyte % and enhanced COX-2 and NOx activity indicate the immunomodulatory potential of BRM in S. aureus SCM. Reduction of TBC could be due to increased leukocytosis or direct microbicidal activity of the activated milk cells. The beneficial effect of the BRM could be used as alternative therapy in the control of S. aureus SCM in cows, either alone or in conjunction with antibiotic therapy.

  5. Over-expression of the bovine FcRn in the mammary gland results in increased IgG levels in both milk and serum of transgenic mice

    PubMed Central

    Lu, Wei; Zhao, Zhihui; Zhao, Yaofeng; Yu, Shuyang; Zhao, Yiqiang; Fan, Baoliang; Kacskovics, Imre; Hammarström, Lennart; Li, Ning

    2007-01-01

    The neonatal Fc receptor (FcRn) protects immunoglobulin G (IgG) from catabolism and is also responsible for IgG absorption in the neonatal small intestine. However, whether it mediates the transfer of IgG from plasma to milk still remains speculative. In the present study, we have generated transgenic mice that over-express the bovine FcRn (bFcRn) in their lactating mammary glands. Significantly increased IgG levels were observed in the sera and milk from transgenic animals, suggesting that the over-expressed bFcRn could bind and protect endogenous mouse IgG and thus extend its lifespan. We also found that injected human IgG showed a significantly longer half-life (7–8 days) in the transgenic mice than in controls (2·9 days). Altogether, the data suggested that bFcRn could bind both mouse and human IgG, showing a cross-species FcRn–IgG binding activity. However, we found no selective accumulation of endogenous mouse IgG or injected bovine IgG in the milk of the transgenic females, supporting a previous hypothesis that IgG was transported from serum to milk in an inverse correlation to its binding affinity to FcRn. PMID:17608809

  6. Hyperimmunised bovine milk and whey: influence of pH and enzymatic treatments on the antigen-binding capacity of immunoglobulin G.

    PubMed

    Dugnol, Julia; Riera, Francisco A

    2016-03-30

    Hyperimmunised bovine milk and whey (whole and defatted) were submitted at 37°C to different pH values (between 1 and 10) and enzymes (pepsin, trypsin and chymotrypsin) at their optimum pH and the IgG immunoactivity against Campylobacter jejuni was measured by means of ELISA assays. The kinetic antigen-binding capacity (ABC) losses follow a hyperbolic-type equation. The ABC of IgG is strongly reduced at low pH (1 and 2) and the effect is lower at alkaline pH (8 and 10). The presence of pepsin (at their optimum pH of 2) almost completely reduced the IgG ABC after 2 h of treatment. The ABCs are higher in whole products (milk and whey). The influence of trypsin and quimotrypsin on the ABCs is moderate (ABC losses lower that 25%). The ABC of IgG obtained from hyperimmunised bovine defatted milk and whey is largely reduced in conditions similar to those found in the human digestive tract. Only whole milk can maintain around 40% of their initial ABC. IgG encapsulation or other methods to protect the immunoglobulin activities could be an alternative to use these type of products in final foods formulae. © 2015 Society of Chemical Industry.

  7. Bovine alpha s1-casein gene sequences direct high level expression of human granulocyte-macrophage colony-stimulating factor in the milk of transgenic mice.

    PubMed

    Uusi-Oukari, M; Hyttinen, J M; Korhonen, V P; Västi, A; Alhonen, L; Jänne, O A; Jänne, J

    1997-01-01

    The generation is reported of transgenic mice expressing human granulocyte-macrophage colony-stimulating factor (GM-CSF) or human erythropoietin (EPO) under the control of bovine alpha s1-casein regulatory sequences. GM-CSF expression was specific to the mammary gland, and levels of human GM-CSF in transgenic mouse milk were in the range of mg ml-1. The specific activity of the milk GM-CSF was similar to that of the recombinant protein produced in Escherichia coli, and the glycosylation-derived size heterogeneity corresponded to that of the native human protein. In spite of the identical bovine regulatory sequences of the fusion genes, the levels of human EPO in transgenic mouse milk were 10(3)-10(6) times lower than those of GM-CSF, ranging from 0.003 to 3 micrograms ml-1. There appeared to be a positive correlation between the amount of EPO in the milk of lactating females and blood haematocrit values. In view of this, other type of constructs should be used to achieve more efficient EPO expression and to circumvent concomitantly-occurring adverse effects. In contrast, the high-level production of recombinant GM-CSF, its resemblance to the native mammalian protein, and mild adverse consequences of transgene expression imply that the current construct could be used for generation of larger GM-CSF transgenic animals to produce this protein in quantities sufficient for therapeutic purposes.

  8. Rapid Communication: MiR-92a as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk.

    PubMed

    Lai, Y C; Fujikawa, T; Ando, T; Kitahara, G; Koiwa, M; Kubota, C; Miura, N

    2017-06-01

    Our aim was to identify a suitable microRNA housekeeping gene for real-time PCR analysis of bovine mastitis-related microRNA in milk. We identified , , and as housekeeping gene candidates on the basis of previous Solexa sequencing results. Threshold cycle (CT) values for , , and did not differ between milk from control cows and milk from mastitis-affected cows. NormFinder software identified as the most stable single housekeeping gene. We evaluated the suitability of the housekeeping gene candidates by using them to assess expression levels of the inflammation-related gene . Regardless of the housekeeping gene candidates used for normalization, relative expression levels of were significantly higher in mastitis-affected samples than in control samples. However, of all the housekeeping genes and gene combinations investigated, normalization with alone generated the difference in relative expression between mastitis-affected and control samples with the highest significance. These results suggest that is suitable for use as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk.

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

    PubMed

    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. Comparison of the dot-immunobinding assay with the serum agglutination test, the rose bengal plate test and the milk ring test for the detection of Brucella antibodies in bovine sera and milk.

    PubMed

    Gürtürk, K; Boynukara, B; Ilhan, Z; Hakki Ekin, I; Gülhan, T

    1999-05-01

    In this study, Brucella antibodies in bovine sera and milk were detected using the dot-immunobinding assay (DIA), the serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT). For this purpose, a total of 116 paired blood and milk samples collected at the same time from 56 aborted and from 60 healthy dairy cows was examined. In DIA, a nitrocellulose membrane (NCM) was used as the solid phase. Antigen adsorbed on the NCM was extracted from Brucella abortus S99 by heat treatment. The results obtained by DIA were compared with those of SAT, RBPT and MRT. Of the 116 paired blood and milk samples, 24 were positive and 72 were negative by all tests used. Serum samples of six aborted cows were positive by DIA, SAT and RBPT but the milk samples were negative by DIA and MRT. Serum and milk samples of four aborted cows gave positive reaction only by DIA tests. The remaining six aborted cows were negative only by MRT and two of them were negative by both RBPT and MRT. Four sera of healthy cows were found to be positive only by SAT.

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

    PubMed

    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.

  12. A Triad of Highly Divergent Polymeric Immunoglobulin Receptor (PIGR) Haplotypes with Major Effect on IgA Concentration in Bovine Milk

    PubMed Central

    Berry, Sarah; Coppieters, Wouter; Davis, Stephen; Burrett, Alayna; Thomas, Natalie; Palmer, David; Kelly, Van; Obolonkin, Vladimir; Sanders, Kathryn; Spelman, Richard; Georges, Michel; Lehnert, Klaus; Snell, Russell

    2013-01-01

    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. PMID:23536764

  13. Development of an analytical method for penicillin G in bovine milk by liquid chromatography with ultraviolet-visible detection and confirmation by mass spectrometric detection.

    PubMed

    Tyczkowska, K; Voyksner, R D; Aronson, A L

    1989-05-05

    A liquid chromatographic (LC) method with ultraviolet-visible photodiode array (UV-VIS PDA) detection was developed to measure penicillin G in bovine milk. A liquid chromatographic-mass spectrometric (LC-MS) procedure was divised to confirm the LC method. The method involved diluting milk with a drug-releasing solvent consisting of acetonitrile-methanol-water and ultrafiltration through a 10,000 dalton cutoff filter. Penicillin G was separated from other components in the ultrafiltrate by ion-paired LC using a reversed-phase microbore column eluted with a 25% acetonitrile solution. The LC method was confirmed by thermospray LC-MS. The detection limit for penicillin G determination in milk was estimated to be 10 ppb for LC with UV-VIS PDA and 100 ppb for LC-MS.

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

    PubMed

    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.

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

    PubMed

    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.

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

    PubMed

    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

  17. Detection of Toxoplasma gondii in raw caprine, ovine, buffalo, bovine, and camel milk using cell cultivation, cat bioassay, capture ELISA, and PCR methods in Iran.

    PubMed

    Dehkordi, Farhad Safarpoor; Borujeni, Mohammad Reza Haghighi; Rahimi, Ebrahim; Abdizadeh, Rahman

    2013-02-01

    This study was conducted to determine the presence of Toxoplasma gondii in animal milk samples in Iran. From a total of 395 dairy herds in three provinces of Iran, 66 bovine, 58 ovine, 54 caprine, 33 buffalo, and 30 camel herds were studied, and from these parts of Iran, 200 bovine, 185 ovine, 180 caprine, 164 buffalo, and 160 camel milk samples were collected from various seasons. Samples were tested for Toxoplasma gondii by cell line culture, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR) technique. Only the results of cell line cultivation were confirmed by bioassay in cat. Results indicated that all herds were infected with Toxoplasma gondii. The culture method showed that 51 out of 889 milk samples (5.73%) were positive for Toxoplasma gondii, and all 51 positive culture results were positive with bioassay in cat. The Fars province had the highest prevalence of Toxoplasma gondii (6.84%). The ELISA test showed that 41 milk samples (4.61%) were positive for the presence of Toxoplasma gondii, while the PCR showed that 46 milk samples were positive for Toxoplasma gondii. The results showed higher sensitivity of PCR and higher specificity of ELISA. Caprine had the highest (10%) and camel had the lowest (3.12%) prevalence rate of parasite. The summer season had the highest (76.47%) but winter (3.92) had the lowest incidence of Toxoplasma gondii. This study is the first prevalence report of direct detection of Toxoplasma gondii in animal milk samples in Iran.

  18. Distinct composition of bovine milk from Jersey and Holstein-Friesian cows with good, poor, or noncoagulation properties as reflected in protein genetic variants and isoforms.

    PubMed

    Jensen, H B; Poulsen, N A; Andersen, K K; Hammershøj, M; Poulsen, H D; Larsen, L B

    2012-12-01

    The objective of this study was to examine variation in overall milk, protein, and mineral composition of bovine milk in relation to rennet-induced coagulation, with the aim of elucidating the underlying causes of milk with impaired coagulation abilities. On the basis of an initial screening of 892 milk samples from 42 herds with Danish Jersey and Holstein-Friesian cows, a subset of 102 samples was selected to represent milk with good, poor, or noncoagulating properties (i.e., samples that within each breed represented the most extremes in regard to coagulation properties). Milk with good coagulation characteristics was defined as milk forming a strong coagulum based on oscillatory rheology, as indicated by high values for maximum coagulum strength (G'(max)) and curd firming rate (CFR) and a short rennet coagulation time. Poorly coagulating milk formed a weak coagulum, with a low G'(max) and CFR and a long rennet coagulation time. Noncoagulating milk was defined as milk that failed to form a coagulum, having G'(max) and CFR values of zero at measurements taken within 1h after addition of rennet. For both breeds, a lower content of total protein, total casein (CN) and κ-CN, and lower levels 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, α(S1)-CN (variants B and C), α(S2)-CN (A), β-CN (A(1), A(2), B, I, and F), κ-CN (A, B, and E), α-lactalbumin (B), and β-lactoglobulin (A, B, and C). In poorly coagulating and noncoagulating milk samples of both breeds, the predominant composite genotype of α(S1)-, β-, and κ-CN was BB-A(2)A(2)-AA, which confirmed a genetic contribution to impaired milk coagulation. Interestingly, subtle variations in posttranslational modification of CN were observed between the

  19. The Intestinal Transport of Bovine Milk Exosomes Is Mediated by Endocytosis in Human Colon Carcinoma Caco-2 Cells and Rat Small Intestinal IEC-6 Cells.

    PubMed

    Wolf, Tovah; Baier, Scott R; Zempleni, Janos

    2015-10-01

    MicroRNAs play essential roles in gene regulation. A substantial fraction of microRNAs in tissues and body fluids is encapsulated in exosomes, thereby conferring protection against degradation and a pathway for intestinal transport. MicroRNAs in cow milk are bioavailable in humans. This research assessed the transport mechanism of bovine milk exosomes, and therefore microRNAs, in human and rodent intestinal cells. The intestinal transport of bovine milk exosomes and microRNAs was assessed using fluorophore-labeled bovine milk exosomes in human colon carcinoma Caco-2 cells and rat small intestinal IEC-6 cells. Transport kinetics and mechanisms were characterized using dose-response studies, inhibitors of vesicle transport, carbohydrate competitors, proteolysis of surface proteins on cells and exosomes, and transepithelial transport in transwell plates. Exosome transport exhibited saturation kinetics at 37°C [Michaelis constant (Km) = 55.5 ± 48.6 μg exosomal protein/200 μL of media; maximal transport rate = 0.083 ± 0.057 ng of exosomal protein · 81,750 cells(-1) · h(-1)] and decreased by 64% when transport was measured at 4°C, consistent with carrier-mediated transport in Caco-2 cells. Exosome uptake decreased by 61-85% under the following conditions compared with controls in Caco-2 cells: removal of exosome and cell surface proteins by proteinase K, inhibition of endocytosis and vesicle trafficking by synthetic inhibitors, and inhibition of glycoprotein binding by carbohydrate competitors. When milk exosomes, at a concentration of 5 times the Km, were added to the upper chamber in transwell plates, Caco-2 cells accumulated miR-29b and miR-200c in the lower chamber, and reverse transport was minor. Transport characteristics were similar in IEC-6 cells and Caco-2 cells, except that substrate affinity and transporter capacity were lower and higher, respectively. The uptake of bovine milk exosomes is mediated by endocytosis and depends on cell and exosome

  20. Size-dependent lipid content of bovine milk fat globule and membrane phospholipids.

    PubMed

    Mesilati-Stahy, Ronit; Mida, Kfir; Argov-Argaman, Nurit

    2011-07-13

    The mammary epithelial cell produces unique structures and a range of diversely sized lipid particles from tens of micrometers to less than 1 μm. The physical, chemical, and biological properties of the differently sized milk fat globules (MFGs) and their complex membranes are not well described. Six size fractions of MFGs were obtained by gravity-based separation and analyzed, and their partial lipidome was determined. The smallest MFGs had a higher concentration of polyunsaturated fatty acids (FAs). The FAs indicative of elongase activity were highest in the smallest MFGs, whereas those FAs indicative of desaturase activity did not differ between size groups. The phosphatidylinositol concentration was highest whereas the phosphatidylserine concentration was lowest in MFGs with an average diameter of 2 μm. Phosphatidylethanolamine and cholesterol concentrations were highest whereas that of sphingomyelin was lowest in MFGs with an average diameter of 3 μm. Phosphatidylcholine concentrations did not vary between the size groups. Results suggest that the assembly of milk fat globules that differ in size is not a homogeneous nor random process and that the differences in composition may reflect discrete biosynthetic routes.

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

    PubMed Central

    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

  2. Development of a semi-nested PCR for the improved detection of Mycoplasma bovis from bovine milk and mucosal samples.

    PubMed

    Hayman, Bernadette; Hirst, Robert

    2003-02-02

    A new forward primer, Mb-F, was designed to improve the sensitivity and reproducibility of the Mycoplasma bovis-specific PCR developed by Ghadersohi et al. [Vet. Microbiol. 56 (1997) 87] for testing clinical samples. A semi-nested (SN) PCR configuration was developed and this provided enhanced sensitivity and reproducibility. The detection limit of the SN PCR was in the range of 10-100cfu/ml and the correct amplicon was amplified from 9.15pg/microliter of total extracted DNA (mixture of M. bovis and bovine cellular DNA). A dot blot assay was also developed and compared with the SN PCR on a number of randomly selected milk and mucosal samples. The dot blot had the same level of detection as the SN PCR. The specificity of the SN configuration was confirmed by Southern blot analysis and automated sequencing of the PCR product. The results from the tests on the samples from cattle, together with those from sheep, provided evidence that M. bovis is host-specific and that most cattle are colonised. The assay was shown to be specific, sensitive and reproducible and could be used successfully to detect M. bovis directly from clinical material without pre-enrichment.

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

    PubMed

    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.

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

    PubMed

    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

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

    PubMed

    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.

  6. Effects of bovine somatotropin on milk yield and composition in Holstein cows in advanced lactation fed low- or high-energy diets.

    PubMed

    Tarazon-Herrera, M A; Huber, J T; Santos, J E; Nussio, L G

    2000-03-01

    The objective of this study was to evaluate the response of Holstein cows to bovine somatotropin (bST) during advanced lactation and its relationship to energy intake. Twenty-four lactating Holstein cows averaging 21 kg of milk/d, and 292 d in milk were assigned to one of three treatment groups in a randomized block design. Blocks were based on the 14 d of pretreatment milk production, and treatment groups were balanced for days in milk. Treatment 1 was a low-energy diet (1.49 Mcal/kg of dry matter) without bST injection; treatment 2 was the low-energy diet plus injection of 500 mg of bST every 14 d; and treatment 3 was a high-energy diet (1.71 Mcal/kg of dry matter) with bST injections as in treatment 2. Treatment was divided into two periods (1 to 49 and 50 to 98 d) to determine if response to bST and energy changed with time on treatment. Results showed that bST significantly (P < 0.05) increased milk, fat-corrected milk, and fat and protein yields; and feed efficiency (fat-corrected milk per dry matter intake) for both periods. Milk yield responses to bST were greater for cows fed the low-than the high-energy diet in both periods. These data show that bST injections for cows in advanced lactation increased performance, but excessive energy diminished the bST response.

  7. Molecular basis of IgE cross-reactivity between human beta-casein and bovine beta-casein, a major allergen of milk.

    PubMed

    Bernard, H; Negroni, L; Chatel, J M; Clement, G; Adel-Patient, K; Peltre, G; Creminon, C; Wal, J M

    2000-01-01

    Twenty patients allergic to cow's milk proteins and with high levels of specific IgE directed against bovine whole casein were selected to evaluate reactivity of their IgE antibodies with human beta-casein. Highly purified human and bovine beta-caseins were prepared by selective precipitations and FPLC separation. Their identity and purity were assessed by HPLC, analysis of amino acid composition, sequencing of the five N-terminal amino acid residues and immunochemical tests. Direct and indirect ELISAs were performed using human and bovine beta-casein coated into microtiter plates and monoclonal anti-human IgE antibody AChE labelled for revelation. Seven sera contained specific IgE directed against human beta-casein. Inhibition studies using native human and bovine beta-caseins as well as bovine beta-casein-derived peptides demonstrated that, depending on the sera, one or several common epitopes located in different parts of the molecule were shared by the two homologous proteins.

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

    PubMed Central

    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

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

    PubMed

    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

  10. The phytanic acid content of the lipids of bovine tissues and milk.

    PubMed

    Lough, A K

    1977-01-01

    In three steers which were given grass silage for six months, the content of phytanic acid (i.e. 3,7,11,15-tetramethyl-hexadecanoic acid) in plasma lipid increased to about 8% of the total fatty acids, whereas after this time the proportion in the total fatty acids of liver and heart lipids was about 1%, and only 0.1% in those of kidney lipids; the acid was present in trace amounts in adipose-tissue triglycerides and was apparently absent from brain lipids. In eight lactating cows which were given grass silage for about 3 months, the content of phytanic acid in the total long chain fatty acids of milk and of plasma was 0.7% and 13%, respectively. In the plasma lipids of both steers and lactating cows, phytanic acid constituted a substantial proportion of the total fatty acids of the triglycerides and phospholipids; the acid was present in lowest proportion in the cholesteryl esters.

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

    PubMed

    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 <6.0. The rate constant for endogenous syneresis was significantly positively

  12. The loss of opsonic activity of bovine milk whey following depletion of IgA.

    PubMed

    Mackie, D P; Meneely, D J; Pollock, D A; Logan, E F

    1986-02-01

    The role of the IgA antibody to Streptococcus agalactiae found in the whey of milks 12 hours after the first intramammary infection of six Friesian first lactation heifers was assessed using an in vitro bactericidal assay. The mean percentage kill of the streptococci by neutrophils in the presence of these wheys was 36.2% while the equivalent figure for the non-infected quarter whey was 0%. When the IgA antibody was absorbed from the infected quarter wheys using class specific IgA antiserum cross linked with glutaraldehyde the percentage kill of the test system fell to 0%. Elution of the absorbed antibody partially restored the activity to a mean percentage kill of 18.2%. The results indicated that the IgA antibody found in infected quarter whey during the acute stages of intramammary infection with Streptococcus agalactiae was responsible for the opsonic activity which pertained at that time.

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

    PubMed Central

    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. PMID:24348176

  14. Prevalence of Bovine Mastitis Pathogens in Bulk Tank Milk in China

    PubMed Central

    Wang, Ya Jing; Qin, Yun; Guix Vallverdú, Roger; Maldonado García, Jaime; Sun, Wei; Li, Shengli; Cao, Zhijun

    2016-01-01

    The objectives of this study were to estimate the herd prevalence of major mastitis pathogens in bulk tank milk (BTM) in China dairy herds, to determine the relationship between the presence of mastitis pathogens and bulk tank milk somatic cell counts (BTSCC), and to investigate the impact of different dairy cattle farming modes and region on bacterial species. BTM samples collected from 894 dairy herds in China were examined for the presence of mastitis pathogens. The Flinders Technology Associates (FTA) cards were used for BTM sample collection, storage, and transportation and bacterial DNA amplification by real-time PCR. Among contagious pathogens, Staphylococcus aureus, Streptococcus agalactiae, and Streptococcus dysgalactiae were detected in 50.1, 92.2, and 72.3% of the 894 BTM samples, respectively. Among environmental pathogens, E. coli, Streptococcus uberis, Enterococcus spp., Klebsiella spp., Serratia marcescens, Corynebacterium bovis, and Arcanobacterium pyogenes were detected in 28.6, 8.9, 35.7, 20.0, 1.3, 17.0, and 67.2% of the BTM samples, respectively. Staphylococcal β-lactamase gene was detected in 61.7% of the BTM samples. The presence of Staphylococcus aureus and Arcanobacterium pyogenes were significantly associated with high BTSCC, respectively. Significant differences were found in presence of Staphylococcus aureus, Streptococcus agalactiae, and Streptococcus dysgalactiae in BTM sampled from the small household farms, dairy-farming communities, and large-scaled dairy farms. There were significant differences in the presence of Streptococcus agalactiae, Streptococcus dysgalactiae, Arcanobacterium pyogenes, staphylococcal β-lactamase gene, Staphylococcus spp., Klebsiella spp., Enterococcus spp., and Streptococcus uberis in BTM among Inner Mongolia, Heilongjiang, and Hebei province. In conclusion, contagious mammary pathogens are predominated among pathogens in BTM samples in China. PMID:27187065

  15. Identification of potent antioxidant bioactive peptides from goat milk proteins.

    PubMed

    Ahmed, Ahmed S; El-Bassiony, Tawfik; Elmalt, Laila M; Ibrahim, Hisham R

    2015-08-01

    Goat milk proteins have gained increasing attention especially the bioactive peptides released from the parent proteins by digestive enzymes. Specifically, the interest in bioactives of goat milk is intensifying due to its reduced allergenicity compared to bovine milk. In this study, proteins of goat milk were fractionated into caseins (GCP) and whey proteins (GWP), hydrolyzed by pepsin and the generated peptides were examined for radical scavenging activities. The hydrolysates of whey (P-GWP) and casein (P-GCP) proteins exhibited potent superoxide anion (O2(・-)) scavenging activity in a dose-dependent manner, as investigated using the natural xanthine/xanthine oxidase (X/XOD) system. The P-GWP and P-GCP dramatically quenched the O2(・-) flux but had negligible effect on the catalytic function of the enzyme, indicating specificity to scavenge O2(・-) but not oxidase inhibition. Further, both P-GWP and P-GCP were able to remarkably quench the chemical DPPH radical. Fractionation of hydrolysates by size-exclusion chromatography produced four fractions (F1-F4) from both hydrolysates, with variable O2(・-) scavenging activities. However, the slow eluting fractions (F4) of both hydrolysates and fast eluting fraction (F2) of P-GCP contained peptides with the highest scavenging activities. Peptides in the active fractions of P-GWP and P-GCP, isolated by reversed phase-HPLC, exhibited significantly strong O2(・-) scavenging activities. MALDI-TOF-MS allowed the identification of several antioxidant peptides derived from both caseins and whey proteins, with β-casein and β-lactoglobulin being the major contributors, respectively. The results demonstrate that digestion with pepsin generates multiple soluble peptides from goat milk protein fractions with remarkable ability to scavenge superoxide radicals and thus providing a fascinating opportunity for their potential candidacy as antioxidant bioactive peptides. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Correlation between mastitis occurrence and the count of microorganisms in bulk raw milk of bovine dairy herds in four selective culture media.

    PubMed

    Souto, Luís I M; Minagawa, Clarice Y; Telles, Evelise O; Garbuglio, Márcio A; Amaku, Marcos; Melville, Priscilla A; Dias, Ricardo A; Sakata, Sonia T; Benites, Nilson R

    2010-02-01

    Milk is the normal secretion of the mammary gland, practically free of colostrum and obtained by the complete milking of one or more healthy animals. Mastitis is an inflammatory process of the mammary gland and it may cause alterations in the milk. The present work aimed to verify whether it is possible, by means of the counts of microorganism in the bulk raw milk in four selective culture media, to establish a correlation with the occurrence of mastitis and therefore, to monitor this disease in bovine dairy herds. The following selective culture media were used: KF Streptococcus Agar, Edwards Agar, Baird-Parker Agar, Blood Agar plus potassium tellurite. Spearman's correlation coefficient was calculated in order to compare the occurrence of mastitis (percentage) in each herd with respective selective culture media counts of microorganisms in bulk raw milk. Thirty-six possibilities were analysed (Tamis and CMT-positive rates were compared with the log-transformed count in four selective culture media) and there was a negative correlation between Tamis 3 and the Baird-Parker Agar plate count. The total results of microbiological tests showed that there were three correlations of the counts in selective culture media. Fifty-two possibilities were analysed and there was a negative correlation between no-bacterial-growth mastitis rates and log10 of KF Streptoccocus Agar plate count and there were two positive correlations between coagulase-positive staphylococci and log10 of Baird-Parker Agar plate count and Blood Agar plus potassium tellurite plate count.

  17. Twinfilin 1 enhances milk bio-synthesis and proliferation of bovine mammary epithelial cells via the mTOR signaling pathway.

    PubMed

    Li, Lu; Liu, Lijie; Qu, Bo; Li, Xueying; Gao, Xuejun; Zhang, Minghui

    2017-10-21

    Twinfilin1 (TWF1) is an actin monomer-binding protein, which biological function has not yet been fully uncovered. In our previous study, we found by mass spectrometry analysis that TWF1 might be one of the major proteins responsible for milk bio-synthesis and proliferation of bovine mammary epithelial cells (BMECs). The purpose of this study was to explore the possible mechanism by which TWF1 regulates signaling pathways that enhance milk bio-synthesis and proliferation of BMECs. We first explored the effects of TWF1 on milk bio-synthesis and cell proliferation, and analyzed the role of TWF1 on the protein levels of signaling molecules (mTOR, SREBP-1c and Cyclin D1) related to milk bio-synthesis and cell proliferation. Then we determinate the impacts of amino acids (methionine and leucine) and hormones (estrogen and prolactin) on the expressions of TWF1. These results reveal that TWF1 is highly induced by the stimulation of amino acids and hormones and involved in regulation of milk bio-synthesis and cell proliferation via the mTOR pathway in BMECs. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    PubMed

    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.

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

    PubMed

    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. Copyright © 2016 American Dairy Science Association. Published by

  20. Different screening tests and milk somatic cell count for the prevalence of subclinical bovine mastitis in Bangladesh.

    PubMed

    Hoque, Md Nazmul; Das, Ziban Chandra; Talukder, Anup Kumar; Alam, Mohammad Shah; Rahman, Abu Nasar Md Aminoor

    2015-01-01

    Identification of cows with subclinical mastitis (SCM) is an important tool for sustainable dairying and implementing effective mastitis control strategies. A total of 892 quarters milk samples from 228 lactating cows were screened by California mastitis test (CMT), White side test (WST), Surf field mastitis test (SFMT), and somatic cell count (SCC) to study the prevalence of bovine SCM in some selected areas of Bangladesh. Out of 228 cows, 148 (64.9%), 138 (60.5%), 132 (57.9%), and 164 (71.9%) were found positive for SCM by CMT, WST, SFMT, and SCC, respectively. The prevalence of bovine SCM was diagnosed 45.7, 40.2, 36.6, and 29.6% in Chittagong, Sirajgonj, Mymensingh, and Gazipur districts, respectively, based on a combination of all tests. The overall quarter-wise prevalence of SCM was 45.7, 43.5, 41.2, and 55.0% for CMT, WST, SFMT, and SCC. Single quarters and left front quarters were more prone to SCM (P < 0.05). Friesian crossbred cows (56.4%), BCS 2.0-2.5 (55.4%), and parity 4-6 (52.4%), the late lactation stage (5-8 months; 64.7%) and high yielding cows (16-20 L/day; 65.3%) were more susceptible to SCM (P < 0.05). The sensitivity of the CMT, WST, SFMT, and SCC was 65.8, 57.9, 51.0, and 82.5%; specificity 76.2, 72.4, 69.5, and 89.4%; percentage accuracy 70.0, 64.8, 59.9, and 85.2%; positive predictive value 75.2, 69.8, 64.9, and 92.7%, respectively. The categories of CMT reactions were strongly correlated with SCC (P < 0.05). Kappa value of SCC was higher than that of other tests (SCC>CMT>WST>SFMT). Thus, CMT was concluded to be the most accurate (r = 0.782) field diagnostic test after laboratory test like SCC (r = 0.924). However, the use of any single test may not be reliable in diagnosing SCM, while the result of CMT supported by SCC might be used effectively to pinpoint diagnosis of SCM in dairy animals than alone.

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

    PubMed

    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.

  2. Determination of naturally occurring progestogens in bovine milk as their oxime derivatives using high performance liquid chromatography-electrospray ionization-tandem mass spectrometry.

    PubMed

    Regal, Patricia; Nebot, Carolina; Vázquez, Beatriz I; Cepeda, Alberto; Fente, Cristina

    2010-08-15

    Hormones and hormone-like substances which are present in the environment have been repeatedly accused of being the cause of most endocrine disruption. However, the possible role of endogenous hormones in food of animal origin deserves to be discussed as well. The relation between steroid hormones and several human health problems has been previously reported, such as prostate and breast cancer, perturbation of human reproduction and endocrine disruption on humans and wildlife. This research is particularly concerned with cow's milk, which contains a considerable amount of sex hormones. A liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous detection and quantification of four naturally occurring steroid hormones in commercial bovine milk (pregnenolone (P5), progesterone (P4), 17-hydroxypregnenolone (17-OHP5), 17-hydroxyprogesterone (P4)). Oxime derivatives of steroids were analyzed in positive ionization and multiple reaction monitoring mode. Methodology has been validated according to Decision 2002/657/EC criteria. This method has been successfully used in real samples. It is fast and easy-handling and provides a useful tool for the assessment of progestogens in bovine milk. Copyright (c) 2010 Society of Chemical Industry.

  3. Visfatin is present in bovine mammary epithelial cells, lactating mammary gland and milk, and its expression is regulated by cAMP pathway.

    PubMed

    Yonezawa, Tomo; Haga, Satoshi; Kobayashi, Yosuke; Takahashi, Tatsuyuki; Obara, Yoshiaki

    2006-12-11

    Visfatin was originally identified as a growth factor for immature B cells, and recently demonstrated to bind insulin receptor. Visfatin mRNA and protein were detected by RT-PCR and Western blot analysis in cloned bovine mammary epithelial cells, lactating bovine mammary gland and human breast cancer cell line, MCF-7. Immunocytochemical staining localized the visfatin protein in the cytosol and nucleus of both cells. Quantitative-RT-PCR analysis revealed that the expression of the visfatin mRNA was significantly elevated when treated with forskolin (500 microM), isopreterenol (1-10 microM) and dibutyric cyclic AMP (1 mM) for 24 h, and significantly reduced when treated with insulin (5-50 ng/ml) and dexsamethasone (0.5-250 nM) for 24 h. These results indicate that mammary epithelial cells express the visfatin protein and secrete them into the milk.

  4. Effect of milk on antibacterial activity of tetracycline against Escherichia coli and Staphylococcus aureus isolated from bovine mastitis.

    PubMed

    Kuang, Ying; Jia, Haoyu; Miyanaga, Kazuhiko; Tanji, Yasunori

    2009-08-01

    The susceptibility of mastitis-causing Escherichia coli and Staphylococcus aureus to two commonly used antibiotics, tetracycline and penicillin G, was tested in raw milk and in Muller-Hinton (MH) broth by introducing a pH indicator, bromocresol purple, which was shown to be a simple, sensitive, and rapid method. The minimum inhibitory concentration (MIC) of penicillin G in milk was the same as those in MH broth, whereas the MIC of tetracycline in milk was 4 to 32 times that in MH. An irreversible binding between tetracycline and large molecules of milk, which might be due to a hydrophobic interaction, was demonstrated by a dialysis test, suggesting the observed impairing effect was due to the action of milk on the tetracycline being tested. Further investigation revealed that much of the reduction of tetracycline's activity in milk was attributable to the milk protein casein, while other heat-sensitive components in milk also play some roles.

  5. Comparison of C18-Carboxypropylbetaine and Glass Bead DNA Extraction Methods for Detection of Mycobacterium bovis in Bovine Milk Samples and Analysis of Samples by PCR

    PubMed Central

    Cornejo, Brandon J.; Sahagún-Ruiz, Alfredo; Suárez-Güemes, Francisco; Thornton, Charles G.; Ficht, Thomas A.; Adams, L. Garry

    1998-01-01

    The purpose of this prospective study was to compare two different milk preparation methods to assay for the presence of Mycobacterium bovis by PCR. Detection by a C18-carboxypropylbetaine (CB-18)-based sample processing method was compared to extraction of DNA from milk with glass beads. Samples from 17 skin test-positive cattle were analyzed. Following CB-18 processing and glass bead extraction, the sensitivity of IS6110-based PCR was 94.1 and 58.8%, respectively (P < 0.025). Because CB-18 processing will permit the proficient use of PCR for diagnosis and surveillance of bovine tuberculosis, it will contribute to the more efficient detection and control of tuberculosis. PMID:9687483

  6. Elucidation of the composition of bovine milk fat triacylglycerols using high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry.

    PubMed

    Mottram, H R; Evershed, R P

    2001-08-17

    Bovine milk fat triacylglycerols (TAGs) have been characterised using high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry (HPLC-APCI-MS) and high-temperature gas chromatography-mass spectrometry (GC-MS). The complex nature of the fat meant that prefractionation was necessary to provide simpler fractions for more detailed molecular analyses. Silica thin-layer chromatography gave rise to two fractions, one of which contained predominantly butyric acid containing TAGs. Gel permeation chromatography (GPC) gave rise to 16 fractions, which were subsequently analysed using HPLC-APCI-MS. Twelve of the GPC fractions were also analysed by high-temperature GC-MS using a capillary column coated with a polarisable stationary phase. TAGs present in the fractions were correlated with those in the chromatogram of the whole milk fat through retention time comparison and the use of mass chromatograms. In total, 120 TAGs were identified.

  7. Prevalence of bovine viral diarrhoea virus antibodies in bulk tank milk of industrial dairy cattle herds in suburb of Mashhad-Iran.

    PubMed

    Garoussi, M Talebkhan; Haghparast, A; Estajee, H

    2008-04-17

    Bulk milk for the presence of antibodies against bovine viral diarrhea virus (BVDV) from 38 industrial dairy cattle herds complexes with 250-3000 Holstein dairy cows in suburb of Mashhad-Iran was tested. None of the herds were vaccinated against BVDV. Commercial indirect ELISA-kit for the detection of specific antibodies was used. The result could be read visually where the optical density (OD) was measured at 450 nm. The percent positivity (PP) values >or=7 and <7 interpreted positive and negative, respectively. According to this study the apparent and the true prevalence of BVDV antibody-positive herds was 89.47 and 93.98%, respectively. The range of PP was 1.59-107.66 among the herds. The OD in 52.63% bulk milk of the herds was very high. It is concluded that exposure to BVD virus was widely distributed in the dairy cattle herds in suburb of Mashhad-Iran.

  8. Genistein effect on xanthine oxidase activity.

    PubMed

    Sumbayev, V V

    2001-01-01

    Genistein was defined to be an allosteric xanthine oxidase inhibitor in the concentrations 0.1-4.0 microM and xanthine oxidase activator with superoxide scavenging activity in the concentrations 5.0 microM and higher. But the most effective allosteric binding with the highest affinity was observed in the genistein concentrations 0.1-1.0 microM. Intraperitoneum injections of genistein (500 micrograms/kg) during three days with the interval 24 hours decrease xanthine oxidase activity in the liver, lung and brain of the Vistar rats.

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

    PubMed

    Shen, Binglei; Zhang, Liying; Lian, Chuanjiang; Lu, Chunyan; Zhang, Yonghong; Pan, Qiqi; Yang, Runjun; Zhao, Zhihui

    2016-02-17

    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.

  10. Sequence Variations in the Bovine Growth Hormone Gene Characterized by Single-Strand Conformation Polymorphism (Sscp) Analysis and Their Association with Milk Production Traits in Holsteins

    PubMed Central

    Yao, J.; Aggrey, S. E.; Zadworny, D.; Hayes, J. F.; Kuhnlein, U.

    1996-01-01

    Sequence variations in the bovine growth hormone (GH) gene were investigated by single strand conformation polymorphism (SSCP) analysis of seven amplified fragments covering almost the entire gene (2.7 kb). SSCPs were detected in four of these fragments and a total of six polymorphisms were found in a sample of 128 Holstein bulls. Two polymorphisms, a T->C transition in the third intron (designated GH4.1) and an A->C transversion in the fifth exon (designated GH6.2), were shown to be associated with milk production traits. GH4.1(c)/GH4.1(c) bulls had higher milk yield than GH4.1(c)/GH4.1(t) (P <= 0.005) and GH4.1(t)/GH4.1(t) (P <= 0.0022) bulls. GH4.1(c)/GH4.1(c) bulls had higher kg fat (P <= 0.0076) and protein (P <= 0.0018) than GH4.1(c)/GH4.1(t) bulls. Similar effects on milk production traits with the GH6.2 polymorphism were observed with the GH6.2(a) allele being the favorable allele. The average effects of the gene substitution for GH4.1 and GH6.2 are similar, with +/-300 kg for milk yield, +/-8 kg for fat content and +/-7 kg for protein content per lactation. The positive association of GH4.1(c) and GH6.2(a) with milk production traits may be useful for improving milk performance in dairy cattle. PMID:8978066

  11. Nuclear Factor of κB1 Is a Key Regulator for the Transcriptional Activation of Milk Synthesis in Bovine Mammary Epithelial Cells.

    PubMed

    Huang, Xin; Zang, Yanli; Zhang, Minghui; Yuan, Xiaohan; Li, Meng; Gao, Xuejun

    2017-02-03

    The nuclear factor of κB (NFκB) family has been well known for its significant role in regulating the expression of numerous genes that control many biological processes. However, it is unclear whether NFκB could regulate milk synthesis. In this study, we identified NFκB1 as a critical regulator for milk synthesis in bovine mammary epithelial cells (BMECs). Gene function study revealed that NFκB1 modulates the expression of mammalian target of rapamycin (mTOR), sterol response element-binding protein (SREBP)-1c, and β4Gal-T2 for milk synthesis. Furthermore, chromatin immunoprecipitation assays showed that both methionine (Met) and estrogen (E) triggered NFκB1 to bind to gene promoters of mTOR, SREBP-1c, and β4Gal-T2 in BMECs. In addition, we confirmed that Met and E triggered NFκB1 expression and phosphorylation via phosphatidylinositol-3-kinase (PI3K) but not mTOR signaling pathway. Taken together, our study reveals that NFκB1 acts as a PI3K but not mTOR-dependent critical mediator for the transcriptional activation of signaling molecules regulating milk synthesis in BMECs.

  12. The regulation of glucose on milk fat synthesis is mediated by the ubiquitin-proteasome system in bovine mammary epithelial cells.

    PubMed

    Liu, Lily; Jiang, Li; Ding, Xiang-dong; Liu, Jian-feng; Zhang, Qin

    2015-09-11

    Glucose as one of the nutrition factors plays a vital role in the regulation of milk fat synthesis. Ubiquitin-proteasome system (UPS) is a vital proteolytic pathway in all eukaryotic cells through timely marking, recognizing and degrading the poly-ubiquitinated protein substrates. Previous studies indicated that UPS plays a considerable role in controlling the triglyceride (TG) synthesis. Therefore, the aim of this study is to confirm the link between high-glucose and UPS and its regulation mechanism on milk fat synthesis in BMEC (bovine mammary epithelial cells). We incubated BMEC with normal (17.5 mm/L) and high-glucose (25 mm/L) with and without proteasome inhibitor epoxomicin and found that, compared with the control (normal glucose and without proteasome inhibitor), both high-glucose concentration and proteasome inhibitor epoxomicin could increase the accumulation of TG and poly-ubiquitinated proteins, and reduce significantly three proteasome activities (chymotrypsin-like, caspase-like, and trypsin-like). In addition, high-glucose concentration combined with proteasome inhibitor further enhanced the increase of the poly-ubiquitinated protein level and the decrease of proteasome activities. Our results suggest that the regulation of high-glucose on milk fat synthesis is mediated by UPS in BMEC, and high-glucose exposure could lead to a hypersensitization of BMEC to UPS inhibition which in turn results in increased milk fat synthesis.

  13. Associations between bovine, human, and raw milk, and beef isolates of non-O157 Shiga toxigenic Escherichia coli within a restricted geographic area of the United States.

    PubMed

    Cobbold, R N; Davis, M A; Rice, D H; Szymanski, M; Tarr, P I; Besser, T E; Hancock, D D

    2008-05-01

    A survey for Shiga toxigenic Escherichia coli in raw milk and beef was conducted within a defined geographic region of the United States. Prevalence rates based on detection of Shiga toxin gene (stx) were 36% for retail beef, 23% for beef carcasses, and 21% for raw milk samples, which were significantly higher than were Shiga toxigenic E. coli isolation rates of 7.5, 5.8, and 3.2%, respectively. Seasonal prevalence differences were significant for stx positivity among ground beef and milk samples. Distribution of stx subtypes among isolates varied according to sample type, with stx1 predominating in milk, stx2 on carcasses, and the combination of both stx1 and stx2 in beef. Ancillary virulence markers eae and ehx were evident in 23 and 15% of isolates, respectively. Pulsed-field gel electrophoresis demonstrated associations between food isolates and sympatric bovine fecal, and human clinical isolates. These data demonstrate that non-O157 Shiga toxigenic E. coli is present in the food chain in the Pacific Northwest, and its risk to health warrants critical assessment.

  14. Quantification of allergenic bovine milk α(S1)-casein in baked goods using an intact ¹⁵N-labeled protein internal standard.

    PubMed

    Newsome, G Asher; Scholl, Peter F

    2013-06-19

    Intact bovine ¹⁵N-α(S1)-casein was used as an internal standard in a selected reaction monitoring (SRM) assay for milk protein in baked food samples containing fats, sugar, and gums. Effects on SRM results of sample matrix composition in two biscuit recipes containing nonfat dry milk (NFDM) were studied, including samples from a milk allergen ELISA proficiency trial. Following extraction of defatted samples with carbohydrate-degrading enzymes and acid precipitation of casein, the SRM assay exhibited an LOQ of <3 ppm NFDM with 60-80% recovery. NFDM levels measured by the SRM assay were 1.7-2.5 times greater than median levels determined by ELISA. Differences were observed in the α(S1)-casein interpeptide SRM ion abundance profile between recipes and after baking. ¹⁵N-α(S1)-Casein increases SRM analysis accuracy by correcting for extraction recovery but does not eliminate underestimation of allergen concentrations due to baking-related milk protein transformation (modifications).

  15. 9 CFR 94.16 - Milk and milk products.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Milk and milk products. 94.16 Section... VESICULAR DISEASE, AND BOVINE SPONGIFORM ENCEPHALOPATHY: PROHIBITED AND RESTRICTED IMPORTATIONS § 94.16 Milk and milk products. (a) The following milk products are exempt from the provisions of this part:...

  16. Xanthine oxidase and xanthine dehydrogenase from an estivating land snail.

    PubMed

    Hermes-Lima, M; Storey, K B

    1995-01-01

    During arousal from estivation in land snails, Otala lactea, active metabolic functions are restored within minutes and oxygen consumption increases dramatically. During the transition from the hypoxic conditions of estivation to normoxia it is possible that xanthine oxidase (XO) in hepatopancreas contributes to the observed lipid peroxidation. Using a fluorometric assay that is based on the oxidation of pterin, the activities and some properties of XO and XO+XDH (sum of XO and xanthine dehydrogenase activities) were measured in hepatopancreas extracts. Km values for pterin for XO and XO+XDH were 9 and 6 microM, respectively, and the Km of XDH for methylene blue was 5 microM. Both XO+XDH and XO activities were inhibited by allopurinol (I50 = 2 microM), pre-incubation at 40 degrees C, and by 5 min H2O2 pre-exposure. Inclusion of azide in the reaction promoted a rise of approximately 70-fold in the inactivation power of H2O2 due to inhibition of high endogenous catalase activity. The I50 for H2O2 of XO+XDH and XO activities in the presence of azide was 0.04 and 0.11 mM, respectively. Unlike the situation for mammalian XO, a previous reduction of O. lactea XO (by pterin) was not necessary to make the enzyme susceptible to H2O2 effects. Interestingly, methylene blue partially prevented both heat- and H2O2-induced inactivation of XO+XDH activity. These data indicate that the formation of an enzyme-methylene blue complex induces protection against heat and oxidative damage at the FAD-active site. Both XO and XO+XDH activites were significantly higher in snails after 35 days of estivation compared with active snails 24 h after arousal from dormancy. The ratio of XO/(XO+XDH) activities was also slightly increased in estivating O. lactea (from 0.07 to 0.09; P < 0.025). XO activity was 0.03 nmol.min-1.mg protein-1 in estivating snails. Compared with hepatopancreas catalase, XO activity is probably too low to contribute significantly to the net generation of oxyradicals, and

  17. Iodine-125-labeled lipoprotein lipase as a tool to detect and study spontaneous lipolysis in bovine milk

    SciTech Connect

    Sundheim, G.; Bengtsson-Olivecrona, G.

    1986-07-01

    The distribution of lipoprotein lipase among cream, casein, and milk serum can be evaluated by addition of a trace amount of /sup 125/I-labeled lipoprotein lipase to milk. Radioactive lipase was distributed in parallel to endogenous lipase under several conditions. In some milk samples, binding of lipase to cream increased when the milk was cooled. Correlation was good between bound labeled lipase and degree of cold-induced lipolysis in corresponding milk samples. Binding of lipase to cream or to casein was not saturable by addition of two-to threefold more lipase than is normally present in milk. In milk with a relatively high fraction of lipase bound to cream, a correspondingly lower fraction was associated with casein, whereas the fraction of lipase in milk serum was similar in all milk samples. Cold-induced binding of lipoprotein lipase to cream was not fully reversed when the milk was warmed again. Heparin released lipase from casein and increased the amount of lipase bound to cream after cooling.

  18. An updated patent review: xanthine oxidase inhibitors for the treatment of hyperuricemia and gout (2011-2015).

    PubMed

    Ojha, Ritu; Singh, Jagjeet; Ojha, Anu; Singh, Harbinder; Sharma, Sahil; Nepali, Kunal

    2017-03-01

    Xanthine oxidase (XO) is a versatile molybdoflavoprotein, widely distributed, occurring in milk, kidney, lung, heart, and vascular endothelium. Catalysis by XO to produce uric acid and reactive oxygen species leads to many diseases. Anti hyperuricemic therapy by xanthine oxidase inhibitors has been mainly employed for the treatment of gout. Area covered: This review covers the patent literature (2011-2015) and also presents the interesting strategies/rational approaches employed for the design of xanthine oxidase inhibitors reported recently. Expert opinion: Recent literature indicates that various non purine scaffolds have been extensively investigated for xanthine oxidase inhibition. The significant potential endowed by heteroaryl based compounds, in particularly fused heterocycles clearly highlights their clinical promise and the need for detailed investigation. Studies by various research groups have also revealed that the flavone framework is open for isosteric replacements and structural modifications for yielding potent non purine xanthine oxidase inhibitors. In addition, various plant extracts recently reported to possess significant xanthine oxidase inhibitory potential presents enough promise to initiate a screening program for the identification of other plant extracts and phytoconstituents possessing inhibitory potential towards the enzyme.

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

    PubMed

    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).

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

    PubMed

    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.

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

    PubMed Central

    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

  2. Kinetic mechanism of chicken liver xanthine dehydrogenase.

    PubMed Central

    Bruguera, P; Lopez-Cabrera, A; Canela, E I

    1988-01-01

    The kinetic behaviour of chicken-liver xanthine dehydrogenase (xanthine/NAD+ oxidoreductase; EC 1.2.1.37) has been studied. Steady-state results, obtained from a wide range of concentrations of substrates and products, were fitted by rational functions of degree 1:1, 1:2, 2:2 and 3:3 with respect to substrates, and 0:1, 1:1, 0:2 and 1:2 with regard to products, using a non-linear regression program which guarantees the fit. The goodness of fit was improved using a computer program that combines model discrimination, parameter refinement and sequential experimental design. The AIC and F tests were also used for model discrimination. For comparative purposes, the xanthine/oxygen oxidoreductase reaction was also studied. From the functions which give the maximum improvement, the complete rate equation was deduced. The significance of the terms was stated by the above methods. It was concluded that xanthine dehydrogenase requires a minimum mechanism of degree 1:1 for xanthine, 2:2 for NAD+, 1:1 for uric acid and 1:2 for NADH in the xanthine/NAD+ oxidoreductase reaction. These are the minimum degrees required but a rate equation of higher degree is not excluded. PMID:3422556

  3. Inactivation and potential reactivation of pathogenic Escherichia coli O157:H7 in bovine milk exposed to three monochromatic ultraviolet UVC lights.

    PubMed

    Yin, Fugui; Zhu, Yan; Koutchma, Tatiana; Gong, Joshua

    2015-08-01

    The ultraviolet (UVC) light irradiation has been recently studied as a novel non-thermal treatment method for milk. However, the potential reactivation of microorganisms following exposure to UVC light in milk medium was not studied yet. Therefore, the present study was conducted to determine the inactivation and reactivation of pathogenic Escherichia coli O157:H7 following exposure to UV light at three monochromatic wavelengths (222, 254 and 282 nm) in bovine milk. The results showed that inactivation of E. coli O157:H7 following exposure to the UV light at 254 nm was higher (P < 0.05) than that following exposure at 222 and 282 nm at the same UV fluence of 5, 10 and 20 mJ/cm(2). The reactivation of E. coli O157:H7 was increased as the incubation time and temperature increased regardless of the UV light sources under dark incubation phases. The evaluated reactivation ratios of E. coli O157:H7 following exposure to the UV light at 254 nm in milk were lower (P < 0.05) than that following exposure at 222 nm after 1 to 6, 2 to 5 and 5-6 h incubation at 4, 20 and 37 °C, respectively. Furthermore, at most incubation time points, the reactivation ratio of E. coli O157:H7 following exposure to these three UV light sources were lower (P < 0.05) than that of non-UV treated cells regardless of the incubation temperature. The lowest reactivation ratios of E. coli O157:H7 were observed after milk exposure to the UV light at 254 nm at 4 °C incubation when compared to that following exposure to the UV light at 222 and 282 nm.

  4. 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.

    PubMed

    Liu, Hongyun; Zhao, Ke; Liu, Jianxin

    2013-01-01

    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.

  5. Simple, rapid determination of enrofloxacin and ciprofloxacin in bovine milk and plasma by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Idowu, Olutosin R; Peggins, James O

    2004-04-01

    A rapid and simple procedure for determination of enrofloxacin and ciprofloxacin in bovine milk and plasma is described. Protein precipitation from both milk and plasma samples was achieved by addition of acetonitrile and phosphoric acid. Acetonitrile was removed with methylene chloride, leaving enrofloxacin and ciprofloxacin in the acidic aqueous extract. The aqueous extract was analyzed by high-performance liquid chromatography (HPLC) with fluorescence detection. The limit of quantitation (LOQ) for enrofloxacin and ciprofloxacin in milk was found to be 2ng/ml. LOQ for enrofloxacin and ciprofloxacin in plasma was found to be 1ng/ml. Linear calibration curves were obtained with correlation coefficient (r(2)) >/=0.99. Analysis of quality control (QC) samples gave results within +/-10% of the nominal values. Inter-assay precision for the analysis of milk QC samples were in the ranges: 4.63-12.49% (for enrofloxacin) and 4.67-9.86% (for ciprofloxacin). Inter-assay precision for the analysis of plasma QC samples were in the ranges: 6.60-17.31% (for enrofloxacin) and 6.14-13.87% (for ciprofloxacin). Intra-assay precision for the analysis of milk QC samples were in the following ranges: 3.65-7.21% (for enrofloxacin) and 1.58-14.28% (for ciprofloxacin). Intra-assay precision for the analysis of plasma QC samples were in the following ranges: 2.17-16.95% (for enrofloxacin) and 3.31-16.31% (for ciprofloxacin). The effectiveness of protein precipitants other than phosphoric acid was investigated. The method described has been applied to a study of the pharmacokinetics of enrofloxacin and ciprofloxacin in lactating dairy cows and beef steers.

  6. Speciation and strain-typing of Staphylococcus agnetis and Staphylococcus hyicus isolated from bovine milk using a novel multiplex PCR and pulsed-field gel electrophoresis.

    PubMed

    Adkins, P R F; Middleton, J R; Calcutt, M J; Stewart, G C; Fox, L K

    2017-03-22

    Staphylococcus hyicus and Staphylococcus agnetis are two coagulase variable staphylococcal species that can be isolated from bovine milk and are difficult to differentiate. The objectives of this study were to characterize isolates of bovine milk origin from a collection that had previously been characterized as coagulase positive S. hyicus based on phenotypic speciation methods and to develop a PCR-based method for differentiating S. hyicus, S. agnetis, and S. aureus. Isolates (n = 62) were selected from a previous study in which milk samples were collected from cows on 15 dairy herds. Isolates were coagulase tested and speciated using housekeeping gene sequencing. A multiplex PCR to differentiate S. hyicus, S. agnetis, and S. aureus was developed. Pulsed-field gel electrophoresis was conducted to strain type isolates. Based on gene sequencing, 44/62 of the isolates were determined to be either S. agnetis (n = 43) or S. hyicus (n = 1). Overall, 88% (37/42) of coagulase positive S. agnetis isolates were found to be coagulase positive at 4 hours. Herd-level prevalence of coagulase positive S. agnetis ranged from 0 to 2.17%. Strain-typing identified 23 different strains. Six strains were identified more than once and from multiple cows within the herd. Three strains were isolated from cows at more than one time point, with 41-264 days between samplings. These data suggest that S. agnetis is likely more prevalent on dairy farms than S. hyicus Also, some S. agnetis isolates in this study appeared to be contagious and associated with persistent infections.

  7. A naturally occurring α(s1)-casein-derived peptide in bovine milk inhibits apoptosis of granulosa cells induced by serum-free conditions.

    PubMed

    Shimizu, T; Ganzorig, K; Miyamoto, A; Ishii, T; Urashima, T; Fukuda, K

    2014-03-01

    Several naturally occurring peptides in bovine milk were characterized by tandem mass spectrometry and Edman degradation. Chromatograms of peptide fractions (passed through an ultra-filtration membrane, nominal molecular weight limit 3000) prepared from colostrum (collected immediately after parturition) and transitional milk (collected 5 days postpartum) showed that they were almost identical. In total, six peptides, α(s1)-CN (f16-23) (RPKHPIKH), α(s1)-CN (f16-24) (RPKHPIKHQ), α(s1)-CN (f17-25) (PKHPIKHQG), α(s1)-CN (f46-52) (VFGKEKV), α(s1)-CN (f94-105) (HIQKEDVPSER), and β-CN (f121-128) (HKEMPFPK), were identified. One of the major peptides, the N-terminal fragment of αs1 -casein, varied structurally during early lactation: α(s1)-CN (f17-25) (PKHPIKHQG) and α(s1)-CN (f16-23) (RPKHPIKH)/α(s1)-CN (f16-24) (RPKHPIKHQ) were found in colostrum and transitional milk, respectively. A chemically synthesized peptide, α(s1)-CN (f16-23) (RPKHPIKH), inhibited apoptosis of bovine granulosa cells induced by serum-free conditions in a dose-dependent manner, in consequence of caspase-3 and caspase-9 suppressions. The physiological function of the peptide remains unclear, but it may have potential use as pharmaceutical agent and as an anti-apoptotic agent in cell culture medium. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.

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

    PubMed Central

    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

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

    PubMed

    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

  10. Fatty acid composition of milk from multiparous Holstein cows treated with bovine somatotropin and fed n-3 fatty acids in early lactation.

    PubMed

    Carriquiry, M; Weber, W J; Dahlen, C R; Lamb, G C; Baumgard, L H; Crooker, B A

    2009-10-01

    Multiparous cows (n = 59) were blocked by expected calving date and previous milk yield and assigned randomly to treatments to determine effects of bovine somatotropin (bST; Posilac, Monsanto Animal Agricultural Group, St. Louis, MO) and source of dietary fat on milk fatty acid composition during the first 140 d in milk. Diets were provided from calving and included whole, high-oil sunflower seeds (SS; 10% of dietary dry matter; n-6/n-3 ratio of 4.6) as a source of linoleic acid or a mixture of Alifet-High Energy and Alifet-Repro (AF; Alifet USA, Cincinnati, OH; 3.5 and 1.5% of dietary dry matter, respectively; n-6/n-3 ratio of 2.6) as a source of protected n-3 fatty acids (15.7% 18:3, 1.3% 20:5, and 1.3% 22:6). Treatments were derived from a 2 x 2 combination of supplemental fat source (SS, AF) and with 0 (SSN, AFN) or 500 (SSY, AFY) mg of bST administered every 10 d from 12 to 70 d in milk and at 14-d intervals thereafter. Milk fatty acid composition was determined in samples collected from 32 cows (8 complete blocks) during wk 2, 8, and 20 of lactation. Data were analyzed as repeated measures using mixed model procedures to determine the effects of diet, bST, week of lactation, and their interactions. Proportions of 18:3 (4.02 vs. 3.59 +/- 0.16%), 20:5 (0.52 vs. 0.41 +/- 0.02%), and 22:6 (0.11 vs. 0.02 +/- 0.02%) were greater and the n-6/n-3 fatty acid ratio (7.40 vs. 8.80 +/- 0.30) was reduced in milk from cows fed AF compared with SS. Proportions of de novo-synthesized fatty acids increased and preformed fatty acids decreased as lactation progressed, but bST administration delayed this shift in origin of milk fatty acids. Transfer efficiency of 18:3, 20:5, and 22:6 from AF to milk fat averaged 36.2, 4.9, and 5.2%, respectively. These efficiencies increased as lactation progressed, but were delayed by bST. Apparent mammary Delta(9)-desaturase activity and milk conjugated linoleic acid (cis-9, trans-11 conjugated linoleic acid) content increased through the

  11. Recombinant bovine somatotropin increases milk yield and calf gain in diverse breeds of beef cattle: associated changes in hormones and indices of metabolism.

    PubMed

    Armstrong, J D; Harvey, R W; Poore, M A; Simpson, R B; Miller, D C; Gregory, G M; Hartnell, G F

    1995-10-01

    In Exp. 1, Angus (A, n = 30), Charolais (C, n = 37), and Simmental (S, n = 30) multiparous cows received (s.c.) recombinantly derived bovine somatotropin (bST; sometribove, 500 mg) or vehicle (VEH) at 2-wk intervals from 124 to 228 d postpartum (DPP). Calves were weaned at 228 DPP. Bovine somatotropin increased (P < .01) milk yield and percentage of milk fat similarly in A, C, and S cows. Calf weaning weight was greater (P < .05) in cows treated with bST than in those given VEH. Administration of bST decreased deposition of fat and increased concentrations of IGF-I, insulin, glucose, and nonesterified fatty acids. In Exp. 2, we compared effects of bST initiated before or after the breeding season. Charolais (n = 33) and S (n = 40) cows were administered (at 2-wk intervals) VEH or bST beginning at 28 DPP (B-bST) or bST beginning at 105 DPP (A-bST). Calves were weaned at 243 DPP. Administration of bST before or after the breeding season increased milk yield on DPP 136 and 194; however, yields were greater in A-bST than in B-bST cows. Milk yields were similar in all cows at 236 DPP, corresponding to decreased forage availability. Calf body weight was greater (P < .05) in A-bST than VEH; B-bST calves were similar to VEH. Fat depth was greater in VEH than in bST-treated cows in C but not in S cows. Serum IGF-I was greater in A- and B-bST than in VEH cows. Mean days from calving to serum progesterone > 1 ng/mL and pregnancy rates were similar in VEH, A-, and B-bST cows. Administration of bST increased cow milk yield and subsequent calf weaning weight when initiated after 100 d postpartum. As anticipated, bST increased IGF-I, insulin, glucose, and nonesterified fatty acids. Administration of bST before and during the breeding season did not affect reproductive performance.

  12. High-performance liquid chromatographic assay for the determination of a semisynthetic avermectin analog (eprinomectin) in bovine milk at parts per billion levels--method development and validation.

    PubMed

    Pollmeier, M; Maier, S; Moriarty, K; DeMontigny, P

    2002-05-25

    A sensitive and automated method has been developed and validated to determine marker residue eprinomectin B(1a) in bovine milk. Extraction of eprinomectin B(1a) from milk is accomplished with acetonitrile after the addition of an internal standard. The extract containing the analytes is evaporated to dryness and reconstituted in a solution containing 30% 1-N-methylimidazole in acetonitrile. Online derivatization is carried out with trifluoroacetic anhydride. Determination of eprinomectin B(1a) and its internal standard is carried out by HPLC using a reversed-phase C(18) column with a mobile phase consisting of methanol, acetonitrile, water, triethylamine and phosphoric acid. The overall extraction recovery of eprinomectin B(1a) is 94% with milk supplemented between 2 and 50 ng/ml eprinomectin B(1a). Precision RSD averaged 3.0% in Laboratory 1 (n=25) compared to 4.3% in Laboratory 2 (n=35). The limit of quantitation is approximately 2 ng/ml eprinomectin B(1a), the limit of detection is approximately 0.25 ng/ml using this method.

  13. Association between the level of antibodies in bulk tank milk and bovine respiratory syncytial virus exposure in the herd

    PubMed Central

    Klem, T. B.; Tollersrud, T.; Østerås, O.; Stokstad, M.

    2014-01-01

    Antibody levels in bulk tank milk (BTM) against bovine respiratory syncytial virus (BRSV) are used to classify BRSV status of herds. The aim of this study was to investigate how these levels correspond with the time at which the herds were infected. Bulk tank milk, individual milk and serum samples from cows and young stock were investigated using an indirect ELISA. Screenings of BTM from 89 dairy herds during two winter seasons revealed a prevalence of positive herds from 82 per cent to 85 per cent. Eleven herds showed a marked increase in antibody levels between two screenings, indicating new infection. However, two of these herds had been free from BRSV for the last five to seven years. Two newly infected herds were monitored for four years and did not appear to get reinfected. Surprisingly, the BTM antibody levels in these herds remained high throughout the study period, but fluctuated significantly. This shows that the levels of antibodies in BTM can remain high for several years, even in herds where reinfection does not occur. BTM serology is a useful tool in the monitoring of infectious diseases in dairy herds, but has limitations as a diagnostic tool for BRSV infections. PMID:24864076

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

    PubMed

    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.

  15. Quantitative analysis of gangliosides in bovine milk and colostrum-based dairy products by ultrahigh performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Lee, Hyeyoung; German, J Bruce; Kjelden, Randy; Lebrilla, Carlito B; Barile, Daniela

    2013-10-09

    Milk gangliosides have gained considerable attention because they participate in diverse biological processes, including neural development, pathogen binding, and activation of the immune system. Herein, we present a quantitative measurement of the gangliosides present in bovine milk and other dairy products and byproducts. Ultrahigh performance liquid chromatography separation was used for high-throughput analysis and achieved a short running time without sacrificing chromatographic resolution. Dynamic multiple reaction monitoring was conducted for 12 transitions for GM3 and 12 transitions for GD3. Transitions to sialic acid fragments (m/z 290.1) were chosen for the quantitation. There was a considerable amount of gangliosides in day 2 milk (GM3, 0.98 mg/L; GD3, 15.2 mg/L) which dramatically decreased at day 15 and day 90. GM3 and GD3 were also analyzed in pooled colostrum, colostrum cream, colostrum butter, and colostrum buttermilk. The separation and analytical approaches here proposed could be integrated into the dairy industry processing adding value to side-streams.

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

    PubMed

    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

    USDA-ARS?s Scientific Manuscript database

    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. Detection and enumeration of Staphylococcus aureus from bovine milk samples by real-time polymerase chain reaction.

    PubMed

    Botaro, B G; Cortinhas, C S; Março, L V; Moreno, J F G; Silva, L F P; Benites, N R; Santos, M V

    2013-01-01

    The aim of this study was to develop and evaluate a real-time quantitative PCR (qPCR)-based method to detect and quantify Staphylococcus aureus in bronopol-preserved milk samples from subclinical intramammary infections (IMI). Serial dilutions of milk artificially inoculated with Staph. aureus ATCC 29213 were used to establish a standard curve (cfu/mL) of the qPCR assay targeting the Staph. aureus thermonuclease-encoding gene nuc according to the strain plate count. The analytical sensitivity, specificity, and repeatability of the qPCR assay were determined. A total of 60 milk samples, collected from mammary quarters without abnormal appearance and with positive isolation of Staph. aureus, were submitted to both the qPCR protocol and Staph. aureus plate counting and results from both methods were compared. Staphylococcus aureus from bronopol-pres