Studies on the Bacillus flora of milk and milk products

Bacillus licheniformis and B. cereus were the most commonly isolated species of Bacillus found in milk at all stages of processing. Bacillus licheniformis was ubiquitous in the farm environment and counts in raw milks heat-treated in the laboratory were higher during the winter months, whilst B. cereus was associated with cattle feed throughout the year, and tended to be more common in raw milks during the summer months. Although B. licheniformis was usually isolated in larger numbers than B. cereus, this pattern changed after raw and pasteurized milks and reconstituted milk powders were pre-incubated at ambient temperatures, and B. cereus came to dominate the Bacillus population, reaching levels associated with enterotoxin production. Investigation of the growth kinetics of strains of both species showed that B. cereus grew faster than B. licheniformis at ambient temperatures. It is suggested that post-pasteurization contamination, which is commonly blamed for spoilage of milk and milk products by B. cereus, is not necessarily the most important source of this organism.     

Immunodetection of added glycomacropeptide in milk formulas and milk powders

The present study aimed the detection of fraudulent manipulation of milk powder with a low cost component--whey powder, by applying the immunochromatographic assay to identify glycomacropeptide. Five commercial milk powder samples of various brands from the national market were analyzed: lactose enriched milk powder type 26, two whole milk powders, vitamin enriched milk powder and full cream milk powder. Our results showed additional whey (1-2%) in 60% of the selected samples after casein removal by precipitation with 20% trichloracetic acid. Another investigated sample--the enriched UHT milk for children aged 4-12 years--proved addition of whey. Other two commercial toddler formula milk powder samples of different brands were used for comparison for the presence of glycomacropeptide. The first sample which was regularly labeled as containing whey protein concentrate was found positive for glycomacropeptide in accordance with the label information, while the second one not containing whey proteins as specified by the product label, was found negative for glycomacropeptide, these two samples being in accordance with the actual legislation.     

Human milk peroxidase is derived from milk leukocytes

Peroxidase enzymes present in human colostrum, saliva, polymorphonuclear leukocytes, and bovine milk were compared with respect to their molecular exclusion chromatographic behavior and immunological cross-reactivity. Human milk peroxidase gave an elution profile similar to myeloperoxidase derived from blood polymorphonuclear leukocytes. Human salivary peroxidase reacted with an antibody directed against bovine lactoperoxidase, but with the same antibody preparation no reaction was detected either with human milk peroxidase or leukocyte myeloperoxidase. We conclude that the peroxidase enzyme in human milk is different from the human salivary and the bovine enzymes and is probably derived from milk leukocytes.     

Zinc binding in cow''s milk and human milk

In both cow's milk and human milk, zinc was associated with proteins of high molecular weight (greater than 100 000), as judged by analysis with Sephadex G-75. Precipitation of the casein at pH 4.6 and filtration of the resultant acid whey on Sephadex G-25 led, however, to the recovery of about 90% of the zinc as a compound of low molecular weight, which was tentatively identified as zinc citrate. Over 95% of the zinc of cow's milk was sedimented with the casein micelles on ultracentrifugation. Filtration of these micellar caseins on Sephadex G-150 gave two peaks containing zinc, which corresponded to aggregates of alpha-casein-kappa-casein and of alpha-casein-beta-casein. Ultracentrifugation of human milk sedimented only approx. 40% of total zinc. Analysis of sediment and supernatant on Sephadex G-150, however, indicated that about 85% of the zinc was associated with a protein complex of molecular weight greater than 150 000. The major protein of this complex was identified as lactoferrin. A minor zinc-binding component of average molecular weight 30 000 was also observed in the supernatant. The results indicated that zinc is bound to different macromolecules in cow's and human milk. This may be a factor affecting the bioavailability to the human infant of zinc from the two milks, and it is suggested that in human milk lactoferrin may be involved in the uptake of zinc.     

Selenium content of milk and milk products of Turkey. II

Selenium content of 1028 milk and milk products of Turkey are presented in this study. The selenium content of human milk (colostrum, transitional, and mature milk), various kinds of milk [cow, sheep, goat, buffalo, paper boxes (3%, 1.5%, 0.012% fat), bottled milk, condensed milk (10% fat), mineral added milk (1.6%), and banana, strawberry, and chocolate milk] and milk products (kefir, yogurt, Ayran, various cheese, coffee cream, ice cream, butter, margarine, milk powder, and fruit yogurt) in Turkey were determined by a spectrofluorometric method. The selenium levels of cow milks collected from 57 cities in Turkey were also determined. Selenium levels in cow milk varied with geographical location in Turkey and were found to be lowest for Van and highest for Aksaray. The results [milk (cow, sheep, goat, buffalo and human) and milks products] were compared with literature data from different countries.     

Temporal changes in milk proteomes reveal developing milk functions

Human milk proteins provide essential nutrition for growth and development, and support a number of vital developmental processes in the neonate. A complete understanding of the possible functions of human milk proteins has been limited by incomplete knowledge of the human milk proteome. In this report, we have analyzed the proteomes of whey from human transitional and mature milk using ion-exchange and SDS-PAGE based protein fractionation methods. With a larger-than-normal sample loading approach, we are able to largely extend human milk proteome to 976 proteins. Among them, 152 proteins are found to render significant regulatory changes between transitional milk and mature milk. We further found that immunoglobulins sIgA and IgM are more abundant in transitional milk, whereas IgG is more abundant in mature milk, suggesting a transformation in defense mechanism from newborns to young infants. Additionally, we report a more comprehensive view of a complement system and associated regulatory apparatus in human milk, demonstrating the presence and function of a system similar to that found in the circulation but prevailed by alternative pathway in complement activation. Proteins involved in various aspects of carbohydrate metabolism are also described, revealing either a transition in milk functionality to accommodate carbohydrate-rich secretions as lactation progresses, or a potentially novel way of looking at the metabolic state of the mammary tissue. Lately, a number of extracellular matrix (ECM) proteins are found to be in higher abundance in transitional milk and may be relevant to the development of infants' gastrointestinal tract in early life. In contrast, the ECM protein fibronectin and several of the actin cytoskeleton proteins that it regulates are more abundant in mature milk, which may indicate the important functional role for milk in regulating reactive oxygen species.     

Peptidome analysis of human skim milk in term and preterm milk

The abundant proteins in human milk have been well characterized and are known to provide nutritional, protective, and developmental advantages to both term and preterm infants. Due to the difficulties associated with detection technology of the peptides, the expression of the peptides present in human milk is not known widely. In recent years, peptidome analysis has received increasing attention. In this report, the analysis of endogenous peptides in human milk was done by mass spectrometry. A method was also developed by our researchers, which can be used in the extraction of peptide from human milk. Analysis of the extracts by LC–MS/MS resulted in the detection of 1000–3000 Da peptide-like features. Out of these, 419 peptides were identified by MS/MS. The identified peptides were found to originate from 34 proteins, of which several have been reported. Analysis of the peptides’ cleavage sites showed that the peptides are cleaved with regulations. This may reflect the protease activity and distribution in human body, and also represent the biological state of the tissue and provide a fresh source for biomarker discovery. Isotope dimethyl labeling analysis was also used to test the effects of premature delivery on milk protein composition in this study. Differences in peptides expression between breast milk in term milk (38–41 weeks gestation) and preterm milk (28–32 weeks gestation) were investigated in this study. 41 Peptides in these two groups were found expressed differently. 23 Peptides were present at higher levels in preterm milk, and 18 were present at higher levels in term milk.     

Hand-rearing of cats using commercial milk powder, Teo milk]

Hysterectomy derived neonatal cats were given three times a day commercial milk diet, Teo milk (SANKYO Co. Ltd.), and showed a good growth comparable to conventional breast-fed cats with high weaning rate (85%) at 5 weeks of age.     

Role of milk secretion in transport of prolactin from blood into milk

In a first experiment prolactin was shown to be uniformly distributed in the milk of dairy cows. Accumulation of prolactin in milk increased with milking interval although milk prolactin content was proportionally greater after 2 h than accumulation of milk. Concentration of milk prolactin and total transfer of prolactin into milk was greater in mammary glands of cows milked just prior to initiation of intravenous infusion of exogenous prolactin. Similarly, inhibition of milk secretion as induced by intramammary infusion of colchicine, markedly reduced milk prolactin concentration and total transfer of prolactin into milk 5h following start of intravenous infusion of exogenous prolactin. Data indicate that transfer of prolactin from blood into milk during established lactation occurs primarily in conjunction with milk secretion.     

Glycobiology of human milk

Glycans are characteristic components of milk, and each species has unique patterns of specific carbohydrates. Human milk is unusually rich in glycans, with the major components being lactose and oligosaccharides, representing approximately 6.8 and 1% of the milk, respectively. Other sources of glycans in human milk include monosaccharides, mucins, glycosaminoglycans, glycoproteins, glycopeptides, and glycolipids. In human milk, the presence and patterns of these glycans vary depending upon the stage of lactation and the maternal genes and their genetic polymorphisms that control glycosyl transferases. The synthesis of milk glycans utilizes a significant portion of the metabolic energy that the mother expends when producing her milk, but other than lactose, these glycans contribute little to the nutritional needs of the infant. The data herein support several functions. 1) Many human milk glycans inhibit pathogens from binding to the intestinal mucosa. 2) Human milk glycans attenuate inflammation. 3) Glycans also directly stimulate the growth of beneficial (mutualist) bacteria of the microbiota (formerly considered commensal microflora of the intestine); these mutualists and their fermentation products can, in turn, (a) inhibit pathogens, (b) modulate signaling and inflammation, and (c) the fermentation products can be absorbed and utilized as a source of dietary calories. These functions can help direct and support intestinal postnatal growth, development, and ontogeny of colonization. The many functions of the milk glycans may synergistically protect infants from disease. Hence, human milk glycans and their homologs may serve as novel prophylactic or therapeutic agents for a diverse range of deleterious conditions.     

Ultrastructural analysis of milk fat globule membranes of colostrum and human milk

Lacteous fat globules with their membranes from human milk and calostrum were studied by scanning and transmission electron microscopy. The first appear as spheroidal structures with some irregularities on the surface. Under the transmission electron microscope these irregularities are composed of islets from a material morphologically similar to cytoplasm, and with structures that resemble a fragment of rugose endoplasmic reticulum. The membranes in specimens fixed immediately after secretion are tri-layered, similar in appearance to those of a single membrane, but in samples fixed between two and four hours after secretion the details are unclear. Through freeze-etching, the laminar aspect of the fat globules is observed.