This study aimed at utilizing ultrasound treatment to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli (Lactobacillus acidophilus BT 1088, L. fermentum BT 8219, L. acidophilus FTDC 8633, L. gasseri FTDC 8131) were treated with ultrasound (30 kHz, 100 W) at different amplitudes (20%, 60% and 100%) for 60, 120 and 180 s prior to inoculation and fermentation in biotin-soymilk. The treatment affected the fatty acids chain of the cellular membrane lipid bilayer, as shown by an increased lipid peroxidation (P<0.05). This led to increased membrane fluidity and subsequently, membrane permeability (P<0.05). The permeabilized cellular membranes had facilitated nutrient internalization and subsequent growth enhancement (P<0.05). Higher amplitudes and longer durations of the treatment promoted growth of lactobacilli in soymilk, with viable counts exceeding 9 log CFU/mL. The intracellular and extracellular β-glucosidase specific activities of lactobacilli were also enhanced (P<0.05) upon ultrasound treatment, leading to increased bioconversion of isoflavones in soymilk, particularly genistin and malonyl genistin to genistein. Results from this study show that ultrasound treatment on lactobacilli cells promotes (P<0.05) the β-glucosidase activity of cells for the benefit of enhanced (P<0.05) isoflavone glucosides bioconversion to bioactive aglycones in soymilk.
The purpose of this paper is to investigate the potential prebiotic properties and proliferation mechanism of fermented milk-derived peptides. In this study, fermented milk-derived polypeptides were obtained by extraction, separation, and purification. The purified peptides were used to culture fecal flora in vitro, and the relative abundance and composition of the flora were analyzed by high-throughput 16S rRNA sequencing technology. The results showed that peptides can promote the proliferation of beneficial bacteria Lactococcus in the intestine and inhibit the proliferation of harmful bacteria Escherichia coli-Shigella. The amino acid sequence of polypeptide components was determined and synthesized in vitro to verify the proliferation of intestinal flora; the proliferation mechanism of peptides on Lactococcus lactis was studied using non-targeted LC-MS metabolomics technology. Five important peptides with molecular weights of 1000-2000 Da were identified by LC-MS: GRP1 (LTEEEK), GRP2 (ENDAPSPVM*K), GRP3 (ITVDDK), GRP4 (EAM*APK) and GRP5 (LPPPEK). The results showed that the peptides could affect the arginine biosynthesis pathway and the amino sugar and nucleotide sugar metabolism of Lactococcus lactis. In addition, the peptides increased the expression of organic acids and their derivatives in Lactococcus lactis. This study provides a research basis for expanding the potential sources of new prebiotics and also opens up a new idea for discovering new prebiotics in vitro.
This study aimed to evaluate the effects of ultrasound on Lactobacillus fermentum BT 8633 in parent and subsequent passages based on their growth and isoflavone bioconversion activities in biotin-supplemented soymilk. The treated cells were also assessed for impact of ultrasound on probiotic properties. The growth of ultrasonicated parent cells increased (P<0.05) by 3.23-9.14% compared to that of the control during fermentation in biotin-soymilk. This was also associated with enhanced intracellular and extracellular (8.4-17.0% and 16.7-49.2%, respectively; P<0.05) β-glucosidase specific activity, leading to increased bioconversion of isoflavones glucosides to aglycones during fermentation in biotin-soymilk compared to that of the control (P<0.05). Such traits may be credited to the reversible permeabilized membrane of ultrasonicated parent cells that have facilitated the transport of molecules across the membrane. The growing characteristics of first, second and third passage of treated cells in biotin-soymilk were similar (P>0.05) to that of the control, where their growth, enzyme and isoflavone bioconversion activities (P>0.05) were comparable. This may be attributed to the temporary permeabilization in the membrane of treated cells. Ultrasound affected probiotic properties of parent L. fermentum, by reducing tolerance ability towards acid (pH 2) and bile; lowering inhibitory activities against selected pathogens and reducing adhesion ability compared to that of the control (P<0.05). The first, second and third passage of treated cells did not exhibit such traits, with the exception of their bile tolerance ability which was inherited to the first passage (P<0.05). Our results suggested that ultrasound could be used to increase bioactivity of biotin-soymilk via fermentation by probiotic L. fermentum FTDC 8633 for the development of functional food.
This study was aimed at an evaluation of the potential inheritance of electroporation effects on Lactobacillus fermentum BT 8219 through to three subsequent subcultures, based on their growth, isoflavone bioconversion activities, and probiotic properties, in biotin-supplemented soymilk. Electroporation was seen to cause cell death immediately after treatment, followed by higher growth than the control during fermentation in biotin-soymilk (P<0.05). This was associated with enhanced intracellular and extracellular beta-glucosidase specific activity, leading to increased bioconversion of isoflavone glucosides to aglycones (P<0.05). The growing characteristics, enzyme, and isoflavone bioconversion activities of the first, second, and third subcultures of treated cells in biotin-soymilk were similar to the control (P>0.05). Electroporation affected the probiotic properties of parent L. fermentum BT 8219, by reducing its tolerance towards acid (pH 2) and bile, lowering its inhibitory activities against selected pathogens, and reducing its ability for adhesion, when compared with the control (P<0.05). The first, second, and third subcultures of the treated cells showed comparable traits with that of the control (P>0.05), with the exception of their bile tolerance ability, which was inherited to the treated cells of the first and second subcultures (P<0.05). Our results suggest that electroporation could be used to increase the bioactivity of biotin-soymilk via fermentation with probiotic L. fermentum BT 8219, with a view towards the development of functional foods.
The objective of this study was to evaluate agricultural wastes as immobilizers for probiotics in liquid foods, such as soy milk. Probiotic strains were initially evaluated for acid and bile tolerance and the ability to produce alpha-galactosidase. Rinds of durian, mangosteen, and jackfruit were dried, ground, and sterilized prior to immobilization of selected strains ( Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and Lactobacillus bulgaricus FTCC 0411). Immobilized cells were inoculated into soy milk, and growth properties were evaluated over 168 h at 37 degrees C. Soy milk containing free cells without agrowastes was used as the control. Immobilized probiotics showed increased growth, greater reduction of stachyose, sucrose, and glucose, higher production of lactic and acetic acids, and lower pH in soy milk compared to the control. The results illustrated that agrowastes could be used for the immobilization of probiotics with enhanced growth, utilization of substrates, and production of organic acids.
The paper describes the rationale and importance of the approaches and methodologies of Participatory Rural Appraisal (PRA) to enable constraint analysis, to understand the complexities of farming systems and to improve integrated dairy productivity. Implicit in this objective is Farming Systems Research (FSR), which focused on cropping systems in the 1970's, with the subsequent addition of animal components. The methodology for FSR involves the following sequential components: site selection, site description and characterization (diagnosis), planning of on-farm research, on-farm testing and validation of alternatives, diffusion of results, and impact assessment. PRA is the development of FSR, which involves the active participation of farmers to identify constraints and plan appropriate solutions. In the Coordinated Research Project (CRP), the approach was adapted to 10 different country situations and led to Economic Opportunity Surveys (EOS) and Diagnostic Surveillance Studies (DSS), allowing the planning and implantation of integrated interventions to improve dairy productivity.
This study analyzed the effect of pH (X1), temperature (X2) and inulin amount (X3) on transformation of isoflavones (daidzin and daidzein) to equol in soymilk fermented with Bifidobacterium spp. All responses significantly (p 0.05) difference between the experimental and predicted values, suggested the suitability of established models in explaining the daidzin and daidzein transformation to equol as a function of pH, temperature and inulin.
This study aimed at utilizing electroporation to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli were treated with different pulsed electric field strength (2.5, 5.0 and 7.5 kV/cm) for 3, 3.5 and 4 ms prior to inoculation and fermentation in biotin-soymilk at 37°C for 24 h. Electroporation triggered structural changes within the cellular membrane of lactobacilli that caused lipid peroxidation (p 9 log CFU/ml after fermentation in biotin-soymilk (p
Assessment of calcium bioavailability from non-dairy foods containing moderate amounts of calcium is especially important in populations that have habitually low dairy consumption. Absorption of calcium from milk and tempeh (a traditional fermented soy product) was compared in a sample of Malay subjects. A randomized, crossover design was utilized to assess calcium absorption in 20 postmenopausal women from either a glass of milk (114 g) or from a meal of tempeh (206 g); each containing 130 mg calcium. At each study of Phase 1 (mid-July) and Phase 2 (mid-August), intravenous (42)Ca and oral (44)Ca were administered and calcium absorption was measured in 24-h urine collections post-dosing; with a 1-month washout period between phases. Absorption of calcium from tempeh did not differ significantly from milk (36.9 +/- 10.6% vs. 34.3 +/- 8.6%, respectively). Due to differences in the calcium content of tempeh, four servings of this product would be needed to get the same amount of absorbed calcium as that obtained from a 4-ounce glass of milk. Tempeh may provide readily available calcium for this population of women at risk for low bone mass.
The aim of this study was to evaluate the effects of fortification and nano-size reduction on calcium absorption and bioavailability of milk powder formula in sham, ovariectomized, and ovariectomized-osteoporosis rats as a menopause and menopause-osteoporosis model. Skim milk powder and skim milk powder fortified with calcium citrate and the suitable doses of inulin, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and vitamins D3, K1, and B6 were formulated based on the North American and Western European recommended dietary allowances. Optimization on cycle and pressure of high-pressure homogenizer was done to produce nano-fortified milk powder. In vivo study demonstrated that fortification and calcium citrate nano-fortified milk powder increased absorption and bioavailability of calcium, as well as bone stiffness and bone strength in sham, ovariectomized, and ovariectomized-osteoporosis rats. This study successfully developed an effective fortified milk powder for food application.
High-producing dairy cows experience a state of negative energy balance in the periparturient period that is partially addressed by increasing the rate of fat and protein mobilization. Previous studies have focused on the rate of fat mobilization, and consequently the rate of protein mobilization has not been well characterized. The objective of this study was therefore to determine the change in indicators of muscle mass during early lactation using ultrasonographic measurement of muscle thickness and changes in plasma creatinine concentration. The maximum thickness of the gluteus medius and longissimus dorsi muscles of 106 Holstein cows (34 primiparous, 72 multiparous) was determined ultrasonographically on d -3, 0, 3, 7, 14, 21, and 28 relative to the day of parturition. Plasma creatinine concentration was measured periodically during the same period. Mixed models analysis and Passing-Bablok regression were used to analyze the data. Gluteus medius thickness, longissimus dorsi loin thickness (LDLT), and longissimus dorsi thoracic thickness (LDTT) were decreased at 28 d postpartum compared with d 3 antepartum. Plasma creatinine concentration was weakly associated with gluteus medius thickness, LDLT, and LDTT (Spearman's rho = 0.31, 0.39, and 0.32, respectively). Plasma creatinine concentration in primiparous and multiparous cows at 28 d postpartum decreased by 0.24 and 0.30 mg/dL, respectively, compared with values 3 d antepartum. We concluded that ultrasonographic measurement of LDLT and LDTT and change in plasma creatinine concentration may provide practical methods for monitoring the rate of protein mobilization in periparturient dairy cows. Ultrasonographic examination of LDLT and LDTT therefore complements ultrasonographic measurement of backfat thickness and may be useful in the evaluation of energy reserve mobilization in periparturient dairy cows.
Two experiments were carried out to evaluate the bactericidal impacts of Bacillus amyloliquefaciens CECT 5940 on the shedding of faecal pathogenic bacteria in dairy calves (Experiment 1) and in adults dogs (experiment 2). In the calves experiment, a completely randomized design was used to investigate the faecal bacteria profile of Holstein dairy calves fed with either pasteurized waste milk (PWM; n = 9) or a formulated non-medicated milk replacer (NMR; n = 9) for 60 d. The NMR containing sodium-butyrate and the active probiotic B. amyloliquefaciens CECT 5940. In the dogs experiment, addition of same probiotic (i.e., B. amyloliquefaciens CECT 5940) was carried out in two stages. The first stage started from day 7-37, and the second from day 44-71. The assessment of faecal score measured on day 22, 37, 42, 57, 71 and 77 to determine the texture of the stools. Calves received PWM consumed (P
The aim of this study was to evaluate the effects of ultraviolet (UV) radiation (ultraviolet A (UVA), ultraviolet B (UVB) and ultraviolet C (UVC) at 30-90 J/m²) on the membrane properties of lactobacilli and bifidobacteria, and their bioconversion of isoflavones in prebiotic-soymilk. UV treatment caused membrane permeabilization and alteration at the acyl chain, polar head and interface region of membrane bilayers via lipid peroxidation. Such alteration subsequently led to decreased (p < 0.05) viability of lactobacilli and bifidobacteria immediately after the treatment. However, the effect was transient where cells treated with UV, particularly UVA, grew better in prebiotic-soymilk than the control upon fermentation at 37°C for 24 h (p < 0.05). In addition, UV treatment also increased (p < 0.05) the intracellular and extracellular β-glucosidase activity of lactobacilli and bifidobacteria. This was accompanied by an increased (p < 0.05) bioconversion of glucosides to bioactive aglycones in prebiotic-soymilk. Our present study illustrated that treatment of lactobacilli and bifidobacteria with UV could develop a fermented prebiotic-soymilk with enhanced bioactivity.
Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. The objective of this study was to evaluate the effects of agrowastes from durian (Durio zibethinus), cempedak (Artocarpus champeden), and mangosteen (Garcinia mangostana) as immobilizers for lactobacilli grown in soymilk. Rinds from the agrowastes were separated from the skin, dried, and ground (150 microm) to form powders and used as immobilizers. Scanning electron microscopy revealed that lactobacilli cells were attached and bound to the surface of the immobilizers. Immobilized cells of Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and L. bulgaricus FTCC 0411 were inoculated into soymilk, stored at room temperature (25 degrees C) and growth properties were evaluated over 168 h. Soymilk inoculated with nonimmobilized cells was used as the control. Utilization of substrates, concentrations of lactic and acetic acids, and changes in pH were evaluated in soymilk over 186 h. Immobilized lactobacilli showed significantly better growth (P < 0.05) compared to the control, accompanied by higher production of lactic and acetic acids in soymilk. Soymilk containing immobilized cells showed greater reduction of soy sugars such as stachyose, raffinose, sucrose, fructose, and glucose compared to the control (P < 0.05).
Ten strains of Lactobacillus were evaluated for their viability in soymilk. Lactobacillus acidophilus ATCC 314, L. acidophilus FTDC 8833, L. acidophilus FTDC 8633 and L. gasseri FTDC 8131 displayed higher viability in soymilk and were thus selected to be evaluated for viability and growth characteristics in soymilk supplemented with B-vitamins. Pour plate analyses showed that the supplementation of all B-vitamins studied promoted the growth of lactobacilli to a viable count exceeding 7 log CFU/ml. alpha-Galactosidase specific activity of lactobacilli as determined spectrophotometrically showed an increase upon supplementation of B-vitamins. High-performance liquid chromatography analyses revealed that this led to increased hydrolysis of soy oligosaccharides and subsequently higher utilization of simple sugars. Production of organic acids as determined via high-performance liquid chromatography also showed an increase, accompanied by a decrease in pH of soymilk. Additionally, the supplementation of B-vitamins also promoted the synthesis of riboflavin and folic acid by lactobacilli in soymilk. Our results indicated that B-vitamin-supplemented soymilk is a good proliferation medium for strains of lactobacilli.
Milk producers in Malaysia make extensive use of crossbred Sahiwal Friesian dairy cattle. These animals have, however, been found susceptible to lactation failure. A survey of cows in an experimental herd of F1 Sahiwal Friesian animals indicated that, in 30% of animals, milk yield decreased to negligible levels within the first 8 weeks post partum. Lactation failure was associated with a progressive increase in the amount of residual milk left in the udder after normal milking. By week 3 of lactation, residual milk volume was significantly greater than that in animals that, based on previous lactation history were not susceptible to lactation failure, and accounted for up to 30% of milk available at the morning milking. The cellular consequences of residual milk accumulation were evident in the activities of acetyl-CoA carboxylase, fatty acid synthetase and galactosyltransferase, key enzyme markers of cellular differentiation, which decreased in glands undergoing lactation failure and were lower than values measured in tissue of control cows. Mammary cell number, estimated by tissue DNA content, was also reduced in animals undergoing lactation failure. These indices of mammary development indicate that lactation failure is the result of premature involution in susceptible animals. Premature involution is a predictable consequence of progressive milk stasis in failing lactation, and attributable to an increase in autocrine feedback by inhibitory milk constituents. The progressive increase in residual milk is, on the other hand, unlikely to be attributable to impaired mammary development. Measurements of milk storage during milk accumulation showed no differences between control and lactation failure cows in the distribution of milk between alveolar and cisternal storage compartments. We conclude that lactation failure in Sahiwal Friesian cows is due to a failure of milk removal, and probably the result of an impaired milk ejection reflex rather than to the glands' milk storage characteristics.
Nutritional interventions are now recommended as strategies to delay Alzheimer's disease (AD) progression. The present study evaluated the neuroprotective effect (anti-inflammation) of lactic acid bacteria (either Lactobacillus fermentum LAB9 or L. casei LABPC) fermented cow's milk (CM) against lipopolysaccharide (LPS)-activated microglial BV2 cells in vitro. The ability of CM-LAB in attenuating memory deficit in LPS-induced mice was also investigated. ICR mice were orally administered with CM-LAB for 28 d before induction of neuroinflammation by LPS. Learning and memory behaviour were assessed using the Morris Water Maze Test. Brain tissues were homogenised for measurement of acetylcholinesterase (AChE), antioxidative, lipid peroxidation (malondialdehyde (MDA)) and nitrosative stress (NO) parameters. Serum was collected for cytokine analysis. CM-LAB9 and CM-LABPC significantly (P < 0·05) decreased NO level but did not affect CD40 expression in vitro. CM-LAB attenuated LPS-induced memory deficit in mice. This was accompanied by significant (P < 0·05) increment of antioxidants (SOD, GSH, GPx) and reduction of MDA, AChE and also pro-inflammatory cytokines. Unfermented cow's milk (UCM) yielded greater cytokine lowering effect than CM-LAB. The present findings suggest that attenuation of LPS-induced neuroinflamation and memory deficit by CM-LAB could be mediated via anti-inflammation through inhibition of AChE and antioxidative activities.
The objective of this study was to evaluate the effects of ultraviolet (UV) radiation (UVB; 90 J/m²) on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of L. casei FTDC 2113. UV radiation significantly enhanced (P < 0.05) the growth of parent cells in mannitol-soymilk fermented at 37°C for 24 h. This had led to an enhanced intracellular and extracellular β-glucosidase activity with a subsequent increase in bioconversion of isoflavones in mannitol-soymilk (P < 0.05). UV radiation also promoted (P < 0.05) the tolerance of parent cells towards acidic condition (pH 2 and 3) and intestinal bile salts (oxgall, taurocholic and cholic acid). In addition, parent treated cells also exhibited better (P < 0.05) adhesion ability to mucin and antimicrobial activity compared to that of the control. All these positive effects of UV radiation were only prevalent in the parent cells without inheritance by first, second and third passage of cells. Although temporary, our results suggested that UV radiation could enhance the bioactive and probiotic potentials of L. casei FTDC 2113, and thus could be applied for the production of probiotic products with enhanced bioactivity.