Physicochemical, Nutritional, and Organoleptic Characterization of a Skimmed Goat Milk Fermented with the Probiotic Strain Lactobacillus plantarum C4.

The benefits of goat milk, fermented milks, and probiotics for the humans are well documented. In this study, a novel fermented goat milk was manufactured with the putative probiotic strain Lactobacillus plantarum C4 together with L. bulgaricus and Streptococcus thermophilus. Ultrafiltration was chosen as the skimmed milk concentration method because it produced the best viscosity and syneresis and a high casein content. The viability rate of all bacterial strains was >107 cfu/mL, even after 5 weeks of storage or after in vitro gastrointestinal digestion, which is especially important for exertion of the probiotic strain functionalities. This fermented milk is also a good source of nutrients, having a low lactose and fat content, high protein proportion, and good mineral concentration. According to these data and the overall acceptability described by panelists, this fermented milk is a healthy dairy product comparable with commercially available fermented milks.

Bioaccessible peptides released by in vitro gastrointestinal digestion of fermented goat milks.

In this study, ultrafiltered goat milks fermented with the classical starter bacteria Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivarus subsp. thermophilus or with the classical starter plus the Lactobacillus plantarum C4 probiotic strain were analyzed using ultra-high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) and/or high performance liquid chromatography-ion trap (HPLC-IT-MS/MS). Partial overlapping of the identified sequences with regard to fermentation culture was observed. Evaluation of the cleavage specificity suggested a lower proteolytic activity of the probiotic strain. Some of the potentially identified peptides had been previously reported as angiotensin-converting enzyme (ACE) inhibitory, antioxidant, and antibacterial and might account for the in vitro activity previously reported for these fermented milks. Simulated digestion of the products was conducted in the presence of a dialysis membrane to retrieve the bioaccessible peptide fraction. Some sequences with reported physiological activity resisted digestion but were found in the non-dialyzable fraction. However, new forms released by digestion, such as the antioxidant αs1-casein 144YFYPQL149, the antihypertensive αs2-casein 90YQKFPQY96, and the antibacterial αs2-casein 165LKKISQ170, were found in the dialyzable fraction of both fermented milks. Moreover, in the fermented milk including the probiotic strain, the k-casein dipeptidyl peptidase IV inhibitor (DPP-IV) 51INNQFLPYPY60 as well as additional ACE inhibitory or antioxidant sequences could be identified. With the aim of anticipating further biological outcomes, quantitative structure activity relationship (QSAR) analysis was applied to the bioaccessible fragments and led to potential ACE inhibitory sequences being proposed. Graphical abstract Ultrafiltered goat milks were fermented with the classical starter bacteria (St) and with St plus the L. plantarum C4 probiotic strain. Samples were analyzed using HPLC-IT-MS/MS and UPLC-Q-TOF-MS/MS. After simulated digestion and dialysis, some of the active sequences remained and new peptides with reported beneficial activities were released.

Antioxidant, ACE-inhibitory and antimicrobial activity of fermented goat milk: activity and physicochemical property relationship of the peptide components.

Increasing evidence on goat milk and the health benefits of its derived products beyond its nutritional value show its potential as a functional food. In this study, goat milk fractions were tested for their total antioxidant capacity using different methods (ORAC, ABTS, DPPH and FRAP), as well as their angiotensin-I-converting-enzyme inhibitory and antimicrobial (against Escherichia coli and Micrococcus luteus) activities. Different whey fractions (whey, cation exchange membrane permeate P and retentate R) of two fermented skimmed goat milks (ultrafiltered goat milk fermented with the classical starter bacteria or with the classical starter plus the Lactobacillus plantarum C4 probiotic strain) were assessed. Additionally, P fractions were divided into two sub-fractions after being passed through a 3 kDa cut-off membrane: (a) the permeate with peptides of MW <3 kDa (P < 3); and (b) the retentate with peptides and proteins of MW >3 kDa (P > 3). No differences in biological activities were observed between the two fermented milks. However, the biological peptides present in the P < 3 fraction showed the highest total antioxidant capacity (for the ORAC assay) and angiotensin-I-converting-enzyme inhibitory activity. Those present in the R fraction showed the highest total antioxidant capacity against ABTS˙+ and DPPH˙ radicals. Some antimicrobial activity against E. coli was observed for the fermented milk containing the probiotic, which could be due to some peptides being released by the probiotic strain. In conclusion, small and non-basic bioactive peptides could be responsible for most of the angiotensin-I-converting-enzyme inhibitory and antioxidant activities. These findings reinforce the potential benefits of the consumption of fermented goat milk in the prevention of cardiovascular diseases associated with oxidative stress and hypertension.

Ultrafiltration of skimmed goat milk increases its nutritional value by concentrating nonfat solids such as proteins, Ca, P, Mg, and Zn.

Goat milk has been reported to possess good nutritional and health-promoting properties. Usually, it must be concentrated before fermented products can be obtained. The aim of this study was to compare physicochemical and nutritional variables among raw (RM), skimmed (SM), and ultrafiltration-concentrated skimmed (UFM) goat milk. The density, acidity, ash, protein, casein, whey protein, Ca, P, Mg, and Zn values were significantly higher in UFM than in RM or SM. Dry extract and fat levels were significantly higher in UFM than in SM, and Mg content was significantly higher in UFM than in RM. Ultrafiltration also increased the solubility of Ca and Mg, changing their distribution in the milk. The higher concentrations of minerals and proteins, especially caseins, increase the nutritional value of UFM, which may therefore be more appropriate for goat milk yogurt manufacturing in comparison to RM or SM.

Study of the effect of different fermenting microorganisms on the Se, Cu, Cr, and Mn contents in fermented goat and cow milks.

The aim of this study was to determine the Se, Cu, Cr, and Mn concentrations of different types of goat- and cow-milk fermented products and evaluate the influence of fermenting bacteria (classical fermenting starters and a probiotic strain) on these concentrations. Atomic absorption spectrometry with hydride generation was used to measure Se and electrothermal atomization to measure Cu, Cr and Mn. Analytical parameters determined in the fermented milks demonstrated that the procedures used were adequate for Se, Cu, Cr, and Mn analyses. Se levels were significantly lower in fermented goat milk products than in fermented cow milk products (p<0.05). Se, Cu, Cr, and Mn levels did not differ as a function of the fermenting bacteria used in commercial fermented goat or cow milks or in the lab-produced goat yoghurt. Given the Se, and Cr intakes for healthy adults, goat and cow yogurts may be important dietary sources.

Does Lactobacillus plantarum or ultrafiltration process improve Ca, Mg, Zn and P bioavailability from fermented goats' milk?

Ca, Mg, Zn and P bioavailability from two experimental ultrafiltered fermented goats' milks (one of them with the probiotic Lactobacillus plantarum and another one without it), and fermented goats' milk samples available in the market were evaluated. Solubility, dialysability and a model combining simulated gastrointestinal digestion and mineral retention, transport and uptake by Caco-2 cells were used to assess bioavailability. The highest Ca, Mg, Zn and P bioavailability values always corresponded to the fermented milk developed by our research group, which could be explained by the effect of milk ultrafiltration. The fermented milk with L. plantarum showed higher Ca retention than the ones without the microorganism, and major Ca uptake when compared to commercial products. This fact could be attributed to a positive effect exerted by the probiotic strain.

The probiotic bacterial strain Lactobacillus fermentum D3 increases in vitro the bioavailability of Ca, P, and Zn in fermented goat milk.

We determined calcium, magnesium, phosphorus and zinc levels in a total of 27 samples of commercial goat- and cow-milk fermented products and 9 samples of a goat-milk fermented product with addition of a probiotic bacterial strain, Lactobacillus fermentum D3, manufactured experimentally by our research group. Atomic absorption spectroscopy with flame atomization and UV/VIS spectrophotometry were used as analytic techniques. The results of an in vitro digestion process showed that the bioavailability of calcium, phosphorus, and zinc was significantly higher in our fermented milk containing the probiotic bacterial strain than it was in commercial goat-milk fermented products. Furthermore, our product showed a significantly higher bioavailability of calcium and zinc compared to goat- and cow-milk fermented products made with other microorganisms. We conclude that, in in vitro assays, strain D3 seems to increase the bioavailability of these minerals and that this new product may constitute a better source of bioavailable minerals compared to other products already on the market.

Duplicate portion sampling combined with spectrophotometric analysis affords the most accurate results when assessing daily dietary phosphorus intake.

The assessment of daily dietary phosphorus (P) intake is a major concern in human nutrition because of its relationship with Ca and Mg metabolism and osteoporosis. Within this context, we hypothesized that several of the methods available for the assessment of daily dietary intake of P are equally accurate and reliable, although few studies have been conducted to confirm this. The aim of this study then was to evaluate daily dietary P intake, which we did by 3 methods: duplicate portion sampling of 108 hospital meals, combined either with spectrophotometric analysis or the use of food composition tables, and 24-hour dietary recall for 3 consecutive days plus the use of food composition tables. The mean P daily dietary intakes found were 1106 ± 221, 1480 ± 221, and 1515 ± 223 mg/d, respectively. Daily dietary intake of P determined by spectrophotometric analysis was significantly lower (P < .001) and closer to dietary reference intakes for adolescents aged from 14 to 18 years (88.5%) and adult subjects (158.1%) compared with the other 2 methods. Duplicate portion sampling with P analysis takes into account the influence of technological and cooking processes on the P content of foods and meals and therefore afforded the most accurate and reliable P daily dietary intakes. The use of referred food composition tables overestimated daily dietary P intake. No adverse effects in relation to P nutrition (deficiencies or toxic effects) were encountered.