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Effects of lactose on colon microbial community structure and function in a four-stage semi-continuous culture system.

A semi-continuous four-channel colon simulator was used to study the effects of lactose for the first time on the growth and fermentation dynamics of colonic microbiota. In six separate simulations, lactose supplementation increased the total SCFA concentration by 3-5 fold as compared with the baseline in the respective vessels. The total bacterial density was inversely correlated with lactic acid production (P = 0.003), while production of butyrate (P = 0.007) and propionate (P = 0.02) correlated with higher numbers of bacteria. A major shift in the microbial community structure in the lactose supplemented vessels was demonstrated by bacterial genomic %G+C-profiling of the total population, where lactose supplementation induced a clearly dominant peak in the bifidobacteria prominent area, %G+C 60-65. The transient shift to increased numbers of bifidobacteria (23-27%) of all bacteria in the first two vessels was also confirmed by the bifidobacteria-specific QPCR-method. In conclusion, lactose produced dramatic changes in microbiota composition and activity as compared with the baseline fermentation.

Base Composition↗

Lactose hydrolysis and formation of galactooligosaccharides by a novel immobilized beta-galactosidase from the thermophilic fungus Talaromyces thermophilus.

beta-Galactosidase from the fungus Talaromyces thermophilus CBS 236.58 was immobilized by covalent attachment onto the insoluble carrier Eupergit C with a high binding efficiency of 95%. Immobilization increased both activity and stability at higher pH values and temperature when compared with the free enzyme. Especially the effect of immobilization on thermostability is notable. This is expressed by the half-lifetime of the activity at 50 degrees C, which was determined to be 8 and 27 h for the free and immobilized enzymes, respectively. Although immobilization did not significantly change kinetic parameters for the substrate lactose, a considerable decrease in the maximum reaction velocity V(max) was observed for the artificial substrate o-nitrophenyl-beta-D-galactopyranoside (oNPG). The hydrolysis of both oNPG and lactose is competitively inhibited by the end products glucose and galactose. However, this inhibition is only very moderate as judged from kinetic analysis with glucose exerting a more pronounced inhibitory effect. It was evident from bioconversion experiments with 20% lactose as substrate, that the immobilized enzyme showed a strong transgalactosylation reaction, resulting in the formation of galactooligosaccharides (GalOS). The maximum yield of GalOS of 34% was obtained when the degree of lactose conversion was roughly 80%. Hence, this immobilized enzyme can be useful both for the cleavage of lactose at elevated temperatures, and the formation of GalOS, prebiotic sugars that have a number of interesting properties for food applications.

Enzyme Activation↗

Optimization of an innovative hollow-fiber process to produce lactose-reduced skim milk.

The research field for applications of lactose hydrolysis has been investigated for several decades. Lactose intolerance, improvement for technical processing of solutions containing lactose, and utilization of lactose in whey are the main topics for development of biotechnological processes. We report here the optimization of a hollow-fiber membrane reactor process for enzymatic lactose hydrolysis. Lactase was circulated abluminally during luminal flow of skim milk. The main problem, the growth of microorganisms in the enzyme solution, was minimized by sterile filtration, ultraviolet irradiation, and temperature adjustment. Based on previous experiments at 23 +/- 2 degrees C, further characterization was carried out at 8 +/- 2 degrees C, 15 +/- 2 degrees C (beta-galactosidase), and 58 +/- 2 degrees C (thermostable beta-glycosidase) varying enzyme activity and flow rates. For a cost-effective process, the parameters 15 +/- 2 degrees C, 240 U/mL of beta-galactosidase, an enzyme solution flow rate of 25 L/h, and a skim milk flow rate of about 9 L/h should be used in order to achieve an aimed productivity of 360 g/(L x h) and to run at conditions for the highest process long-term stability.

Animals↗

Induction of mammary prolactin receptors and lactose synthesis after ovariectomy in the pregnant mouse.

The development of prolactin receptors in the mammary gland after ovariectomy was investigated in pregnant KA mice. Mice were ovariectomized on day 13 of pregnancy and used for the determination of the amount of specific binding of 125I-labelled prolactin to the mammary tissue, and the contents of lactose and nucleic acids in the mammary gland 0, 8, 24, and 72 hr after the operation. The specific binding of 125I-labelled prolactin, lactose and RNA contents in the mammary gland remained low until 8 hr, sharply increased 24 hr and decreased 72 hr after ovariectomy. When ovariectomized mice were treated with 0.2 mg progesterone, pregnancy was maintained and an increase (1.5-fold) in the amount of specific binding was observed with an increase of lactose content. Five mg progesterone completely inhibited lactose synthesis. Cortisol administered with progesterone did not show any specific change at the dose used (0.5 to 10 mg). Although the amount of specific binding was also increased after hysterectomy, this increase (2-fold) did not fully cover the increase after ovariectomy (3-fold). These results suggest that the recepter site for prolactin is induced before the initiation of lactose synthesis caused by ovariectomy during pregnancy.

Animals↗

In silico evolved lac operons exhibit bistability for artificial inducers, but not for lactose.

Bistability in the lac operon of Escherichia coli has been widely studied, both experimentally and theoretically. Experimentally, bistability has been observed when E. coli is induced by an artificial, nonmetabolizable, inducer. However, if the lac operon is induced with lactose, the natural inducer, bistability has not been demonstrated. We derive an analytical expression that can predict the occurrence of bistability both for artificial inducers and lactose. We find very different conditions for bistability in the two cases. Indeed, for artificial inducers bistability is predicted, but for lactose the condition for bistability is much more difficult to satisfy. Moreover, we demonstrate that in silico evolution of the lac operon generates an operon that avoids bistability with respect to lactose, but does exhibit bistability with respect to artificial inducers. The activity of this evolved operon strikingly resembles the experimentally observed activity of the operon. Thus our computational experiments suggest that the wild-type lac operon, which regulates lactose metabolism, is not a bistable switch. Nevertheless, for engineering purposes, this operon can be used as a bistable switch with artificial inducers.

Computer Simulation↗

Lactose intolerance in infants, children, and adolescents.

The American Academy of Pediatrics Committee on Nutrition presents an updated review of lactose intolerance in infants, children, and adolescents. Differences between primary, secondary, congenital, and developmental lactase deficiency that may result in lactose intolerance are discussed. Children with suspected lactose intolerance can be assessed clinically by dietary lactose elimination or by tests including noninvasive hydrogen breath testing or invasive intestinal biopsy determination of lactase (and other disaccharidase) concentrations. Treatment consists of use of lactase-treated dairy products or oral lactase supplementation, limitation of lactose-containing foods, or dairy elimination. The American Academy of Pediatrics supports use of dairy foods as an important source of calcium for bone mineral health and of other nutrients that facilitate growth in children and adolescents. If dairy products are eliminated, other dietary sources of calcium or calcium supplements need to be provided.

Adolescent↗

Kinetics of plasma fructose and glucose when lactose and fructose are used as energy supplements for neonatal calves.

Shortly after birth, plasma glucose and fructose concentrations of the neonate decline and thus leave blood sugar below the homeostatic mode. Two trials were conducted to determine the plasma glucose and fructose kinetics in control and supplemented calves for 108 h after birth. In the short-term trial, six Holstein calves were given 40 g of either fructose, lactose, or water (control) orally at 1 and 96 h after birth. Treatments were administered with a colostrum substitute (Life Boost) at 1 h and whole milk at 96 h. Rectal temperatures and changes in plasma glucose and fructose concentrations were monitored at close intervals for 12 h after supplementation. In the long-term trial, 15 Holstein calves were given 40 g of either lactose, fructose, or water (control) at 1 h after birth and at 12-h intervals for 81 h. Plasma glucose and fructose concentrations were determined before and 4 h after each of the seven feedings. Early postpartal feeding of fructose suppressed plasma glucose (approximately 50%), with a reciprocal rise in plasma fructose. Irrespective of treatment, plasma glucose concentrations did not stabilize (approximately 100 mg/dL) until 17 to 24 h after birth. After 24 h, lactose supplements increased concentrations of plasma glucose 4 h after supplementation (169.7 +/- 8.2 mg/dL), compared with those in calves that did not receive the additional lactose. After 24 h, fructose supplements did not affect plasma glucose, but plasma fructose concentrations increased (82.6 +/- 12.4 mg/dL) 4 h after administration. The response to fructose supplements declined by 11.4 mg x dL(-1) x d(-1). Fructose was not detected in the plasma of control or lactose-treated calves after 17 h after birth. Calves that received fructose supplements had rectal temperatures 8 and 10 h after birth that were higher than those of the other calves. The mechanisms of sugar metabolism change quickly following birth. Oral sugar supplements increase the total plasma sugar concentrations of treated calves.

Animals↗

Clinical aspects of lactose intolerance in children and adults.

The principal carbohydrate of human milk is the disaccharide lactose. In human and all mammalian species, lactose is hydrolyzed in the small intestine by lactase-phlorizin hydrolase, also abbreviated as lactase. The absence of lactase results in the passage of undigested lactose into the large intestine and is associated with a well-known clinical syndrome: lactose intolerance. Low lactase levels result either from intestinal injury or, as in the majority of world's adult population, from alterations in the genetic expression of lactase. In this review terminology, pathophysiology, symptoms, diagnostic procedures, and therapy of lactose intolerance will be discussed.

Adult↗

Inhibition of lactose transport in E. coli by N-ethylmaleimide-beta-galactoside, N-ethylsuccinimide-beta-galactoside and N-ethylmaleimide.

N-ethylmaleimide (NEM) inhibits lactose uptake in E. coli by reacting with the M protein component of the lac permease system. In an attempt to estimate the distance between the NEM reactive site and the substrate binding site, we have synthesized a beta-galactoside with NEM as the aglycon moiety (NEM-gal). NEM-gal was a more effective inhibitor of lactose transport than was NEM. Part of the inhibition by NEM-gal was caused by competition with lactose for the substrate binding site. To estimate this part of the inhibition, we synthesized the saturated and thus the unreactive N-ethylsuccinimide (NES) analog of NEM-gal. Nes-gal was a competitive inhibitor of lactose uptake. The remainder of the inhibition by NEM-gal followed first-order kinetics with the same rate constant as NEM. In addition, the protective effect of thiodigalactoside against the inhibition of transport by NEM was also observed against irreversible inhibition by NEM-gal. We suggest that the reactivity of NEM was unaltered by bringing it near the beta-galactoside binding site by way of covalent attachment to galactose. We conclude that the distance between the NEM reactive site and the position of the glycosidic oxygen of beta-galactosides bound to the lactose site is greater than 8 A.

Biological Transport↗

Detection of glucose, galactose, and lactose in milk with a microdialysis-coupled flow injection amperometric sensor.

A microdialysis-coupled flow injection amperometric Sensor (microFIAS) was used to determine glucose, galactose, and lactose in milk. The sensor is based on enzyme-catalyzed reaction in combination with the three well-established analytical techniques, namely; microdialysis sampling, flow injection analysis (FIA), and amperometric detection. With the multianalyte sensor it was possible to detect glucose and galactose by sequential injection of their corresponding oxidase enzymes: glucose oxidase and galactose oxidase, while lactose was determined by injection of a mixture of beta-galactosidase and glucose oxidase enzymes. The sensor showed a linear response between 0.05 and 10 mM for glucose, between 0.1 and 20 mM for galactose and between 0.2 and 20 mM for lactose, respectively. The relative standard deviation values of the sensor measurements for glucose, galactose, and lactose were 3-4% (n = 3). The sensor measurements for lactose content in milk were compared with a standard method with an infrared spectrophotometer.

Animals↗

Enhanced lactose cheese milk does not guarantee calcium lactate crystals in finished cheddar cheese.

Three experimental batches of Cheddar cheese were manufactured in duplicate, with standardization of the initial cheese-milk lactose content to high (5.24%), normal (4.72%, control), and low lactose (3.81%). After 35 d of aging at 4.4 degrees C, the cheeses were subjected to temperature abuse (24 h at 21 degrees C, unopened) and contamination (24 h at 21 degrees C, packages opened and cheeses contaminated with crystal-containing cheese). After aging for 167 d, residual cheese lactose (0.08 to 0.43%) and L(+)-lactate concentrations (1.37 to 1.60%) were high and D(-)-lactate concentrations were low (<0.03%) for all cheeses. No significant differences in lactose concentrations were attributable to temperature abuse or contamination. No significant differences in L(+)- or D(-)-lactate concentrations were attributable to temperature abuse. However, concentrations of L(+)-lactate were significantly lower and D(-)-lactate were significantly higher in contaminated cheeses than in control cheeses, indicating inoculation (at d 35) with heterofermentative nonstarter lactic acid bacteria able to racemize L(+)-lactate to D(-)-lactate. The fact that none of the cheeses exhibited crystals after 167 d demonstrates that high cheese milk or residual lactose concentrations do not guarantee crystal formation. Contamination with nonstarter lactic acid bacteria can significantly contribute to D(-)-lactate accumulation in cheese.

Animals↗

Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on cheddar cheese quality: pH changes during ripening.

The pH of cheese is an important attribute that influences its quality. Substantial changes in cheese pH are often observed during ripening. A combined effect of calcium, phosphorus, residual lactose, and salt-to-moisture ratio (S/M) of the cheese on the changes in cheese pH during ripening was investigated. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), lactose at pressing (2.4 vs. 0.78%), and S/M (6.4 vs. 4.8%) were manufactured. All the cheeses were salted at a pH of 5.4, pressed for 5 h, and then ripened at 6 to 8 degrees C. The pH of the salted curds before pressing and the cheeses during 48 wk of ripening was measured. Also, cheeses were analyzed for water-soluble Ca and P, organic P, and bound inorganic P during ripening. Changes in organic acids' concentration and shifts in the distribution of Ca and P between different forms were studied in relation to changes in pH. Cheeses with low S/M exhibited a larger increase in acid production during ripening compared with high S/M cheeses. Cheeses with the highest concentration of bound inorganic P exhibited the highest pH, whereas cheeses with the lowest concentration of bound inorganic P exhibited the lowest pH among the 8 treatments. Although conversion of lactose to short-chain, water-soluble organic acids decreased cheese pH, bound inorganic phosphate buffered the changes in cheese pH. Production of acid in excess of the buffering capacity (which was the case in low Ca and P and low S/M treatments) led to a low pH, whereas solubilization of bound inorganic P in excess to acid production (which was the case in high Ca and P and high S/M treatments) led to an increase in pH. However, for cheeses with high Ca and P and low S/M, changes in cheese pH were influenced by the level of residual lactose. Hence, pH changes in Cheddar cheese can be modulated by a concomitant control on the amount and state of Ca and P, level of residual lactose, and S/M of the cheese.

Acids↗

Lactobacillus acidophilus as a dietary adjunct for milk to aid lactose digestion in humans.

The effect of Lactobacillus acidophilus on lactose utilization of humans was determined by our comparing the amount of hydrogen excreted in their breath after consuming milk containing either 0, 2.5 X 10(6), 2.5 X 10(7), or 2.5 X 10(8) Lactobacillus acidophilus/ml daily for 6 days. Consumption of milk without cells of Lactobacillus acidophilus for 1 wk did not affect lactose utilization. Milk containing either 2.5 X 10(6) or 2.5 X 10(8) Lactobacillus acidophilus/ml improved lactose utilization. However, consumption of milk containing an intermediate 2.5 X 10(7)/ml did not improve utilization based on comparison of group means before and after consuming the test milk. The lack of a significant effect for the latter group of test subjects was probably due to large increases of excreted hydrogen on day 7 as compared to day 0 by two of the six test subjects. The beneficial effect of Lactobacillus acidophilus was immediate and did not require that milk be consumed daily. Improved digestion of lactose was not due to hydrolysis of the lactose prior to consumption, which indicated that the beneficial effect must have occurred in the digestive tract after consumption of milk containing L. acidophilus.

Adult↗

Genetic parameters for yield traits of Holsteins, including lactose and somatic cell score.

First lactation milk, fat, protein, and lactose yields and percentage yields were analyzed using a multiple-trait sire model including herd-year-season, sire group, and age of cow as fixed effects. Somatic cell score was fit both as a fixed effect in the model and as an additional dependent variable in two analyses with almost identical results. Variance components were estimated using REML with an expectation-maximization algorithm including sire relationships. Lactose means ranged from 4.84 to 4.97% across three dairy breeds. Data used to estimate variance components were first lactation Holstein records collected from 1986 to 1988 from 5246 daughters of 392 AI sires. Heritability estimates were .30, .29, .27, and .26 for milk, fat, protein, and lactose yields; .45, .47, and .53 for fat, protein, and lactose percentage yields; and .16 for somatic cell score. Genetic correlations of lactose percentage with milk, fat, protein, fat and protein percentages, and somatic cell score were -.30, -.16, -.21, .10, .29, and -.11, respectively, and phenotypic correlations were -.08, -.02, .01, .11, .29, and -.15.

Animals↗

Effect of lactose on calcium absorption from the rat small intestine with a non-flushed ligated loop.

To assess the effect of dietary lactose on the utilization of 40Ca and 45Ca, a one-day balance experiment was carried out. The urinary 45Ca excretion and 45Ca deposition in the femur were significantly higher in the 10% lactose diet group. In the second experiment, non-flushed lower small intestine was ligated in situ and calcium absorption was observed shortly after injecting lactose solution into the loop during the postprandial state. The estimated amounts of 40Ca and 45Ca absorbed and of 45Ca deposited in the femur were increased in the presence of lactose in the test solution. These results suggest that dietary lactose stimulates calcium utilization and also increases calcium absorption from the undisturbed small intestine containing digests.

Animals↗

Temporal effect of prolactin on the activities of lactose synthetase, alpha-lactalbumin, and galactosyl transferase in mouse mammary gland explants.

The onset of the prolactin (PRL) stimulation of lactose synthesis is between 4 and 8 hr after adding PRL to cultured mouse mammary tissues. The synthesis of lactose is catalyzed by the enzyme lactose synthetase, which is composed of two parts, alpha-lactalbumin and galactosyl transferase. In time-sequence studies, it was found that the activity of galactosyl transferase is enhanced by PRL in concert with the onset of the PRL stimulation of lactose synthesis. In contrast, the earliest detectable effect of PRL on alpha-lactalbumin activity occurred 24 hr after adding PRL to the cultures. It is, therefore, apparent that the rate-limiting component for the PRL stimulation of lactose synthesis in cultured mouse mammary tissues is galactosyl transferase activity.

Animals↗

Effect of dietary lactose and cell concentration on the ability of a continuous-flow-derived bacterial culture to control Salmonella cecal colonization in broiler chickens.

The effect of dietary lactose and of cell concentration of a continuous-flow (CF) derived bacterial culture on Salmonella typhimurium cecal colonization in 10-d-old broiler chickens was examined. One-day-old chicks were provided 1) no CF culture and the control diet; 2) no culture and a 2% lactose diet; 3) CF culture (10(8) or 10(11) anaerobic cfu) and control diet; or 4) CF culture (10(8) or 10(11) anaerobic cfu) and 2 to 4% lactose diet. All groups were challenged orally with 10(4) S. typhimurium at 3 d of age. Salmonella typhimurium growth in cecal contents was significantly decreased (P < .05) at 10 d of age when 2% lactose was used in combination with CF culture containing 10(8) or 10(11) anaerobic cfu. The protection factor (log10 Salmonella control diet divided by log10 Salmonella treatment group) for these treatment groups was 2.49 and 9.26, respectively. Results indicate that birds treated with CF culture and maintained on 2% dietary lactose are protected against S. typhimurium colonization. Additionally, inoculating birds with CF culture containing a higher concentration of anaerobic colony-forming units resulted in a substantially higher protection factor.

Animals↗

Management and treatment of lactose malabsorption.

Lactose malabsorption is a very common condition characterized by intestinal lactase deficiency. Primary lactose malabsorption is an inherited deficit present in the majority of the world's population, while secondary hypolactasia can be the consequence of an intestinal disease. The presence of malabsorbed lactose in the colonic lumen causes gastrointestinal symptoms. The condition is known as lactose intolerance. In patients with lactase nonpersistence, treatment should be considered exclusively if intolerance symptoms are present. In the absence of guidelines, the common therapeutic approach tends to exclude milk and dairy products from the diet. However, this strategy may have serious nutritional disadvantages. Several studies have been carried out to find alternative approaches, such as exogenous beta-galactosidase, yogurt and probiotics for their bacterial lactase activity, pharmacological and non pharmacological strategies that can prolong contact time between enzyme and substrate delaying gastrointestinal transit time, and chronic lactose ingestion to enhance colonic adaptation. In this review the usefulness of these approaches is discussed and a therapeutic management with a flow chart is proposed.

Adaptation, Physiological↗