Different Ultrasound Exposure Times Influence the Physicochemical and Microbial Quality Properties in Probiotic Goat Milk Yogurt.

Despite goat milk having health benefits over cow milk, goat milk yogurt (GY) presents low consistency and viscosity, which reduces its overall acceptability by the consumer. Thus, new innovative methods can be an alternative to improve the quality of GY. Hence, this study aimed to investigate the effect of ultrasound (US) treatment with different sonication times on quality parameters of probiotic GY during refrigerated storage. US treatment was conducted at 20 KHz for 3, 6, and 9 min in yogurt. Lactobacillus bulgaricus and Lactobacillus acidophilus LA-5 were sensitive to US treatment, presenting a decrease in the yogurts stocked. This loss of viability led to reduced post-acidification due to smaller lactose metabolization in yogurt samples submitted to the US. Among tested treatments, the application of 6 min enhanced the apparent viscosity and consistency index of GY yogurts. In addition, this time also reduced tyramine and total biogenic amine (BAs) content. These findings suggest that 6 min of sonication is a promising way to improve the rheological properties and reduce the acidity and BAs content in GY. Further studies should be performed to optimize the US setting conditions to preserve the probiotic culture viability in yogurts.

Low-Dose Electron-Beam Irradiation for the Improvement of Biofilm Formation by Probiotic Lactobacilli.

The effects of 50-150 gray electron-beam irradiation on the biofilm-formation ability and cell surface hydrophobicity of the commercial strain, Lactobacillus acidophilus DDS®-1, from Lacto-G (a marketed synbiotic formulation) and the putative probiotic, L. rhamnosus Vahe, were evaluated. No significant changes in cell surface hydrophobicity were found after irradiation, while increases in biofilm-formation abilities were documented for both investigated microorganisms 0.22 ± 0.03 vs. 0.149 ± 0.02 (L. rhamnosus Vahe, 150 Gy) and 0.218 ± 0.021 vs. 0.17 ± 0.012 (L. acidophilus DDS®-1, 150 Gy). Given this, the use of electron-beam irradiation (50-100 Gy) for the treatment of L. rhamnosus Vahe and L. acidophilus DDS®-1 cells may be considered in product sterilization, quality improvement, and packaging practices.

The survival of irradiated lactobacilli in the simulated gastrointestinal conditions with antibiotic ceftazidime.

Lactobacillus acidophilus is one of the widespread probiotic bacteria that can overcome acid and bile barrier of stomach and intestine, respectively, and then have beneficial effects on the host improving its intestinal microbial balance. The cell membrane FO F1 -ATPase is an important factor in the response and tolerance to low pH through the action of controlling the H+ concentration between the cell cytoplasm and external medium. In this study, the effects of extremely high-frequency EMI at the frequencies of 51·8 GHz and 53 GHz and cetfazidime ( µmol l-1 ) on survival of L. acidophilus VKM B-1660 in the gastrointestinal model in vitro and on ATPase activity of their membrane vesicles were investigated. Irradiated L. acidophilus survived in media with acid pH; the irradiation stimulated N,N'-dicyclohexylcarbodiimide-sensitive FO F1 -ATPase activity under acidic conditions, but enhanced the inhibitory effects of ceftazidime. Probably irradiated L. acidophilus is overcoming the acid barrier even in the presence of ceftazidime due to the FO F1 -ATPase. The obtained results can allow the use of L. acidophilus in food industry, veterinary and medicine. SIGNIFICANCE AND IMPACT OF THE STUDY: The probiotic property of lactobacilli is defined with survival in different conditions of human digestive tract even in the presence of antibiotics and subjected to electromagnetic irradiation (EMI) at the extremely high frequency. Despite the fact that EMI and antibiotic ceftazidime affected Lactobacillus acidophilus; the viable number of bacterial cells was decreased in in vitro gastrointestinal model, but they could to grow in fresh growth medium. The changes in the FO F1 -ATPase activity were obtained at acidic pH. Thus, these bacteria can overcome acid barrier due to the FO F1 -ATPase: the irradiation stimulates the FO F1 -ATPase activity in the acidic conditions, but enhances the effects of ceftazidime. The results are important for identifying the mechanisms of lactobacilli survival for physical and chemical factors and valuable for use.

Strategic ultrasound-induced stress response of lactic acid bacteria on enhancement of ß-glucosidase activity for bioconversion of isoflavones in soymilk.

The purpose of the study is to develop the new methodology of strategic ultrasound treatment on lactic acid bacteria (LAB) to induce stress response for the enhancement of ß-glucosidase activity that can be used for the biotransformation of glucosides into aglycones isoflavones in soymilk. Among the five LAB tested, Lactobacillus acidophilus BCRC 10695 showed the best ability to release ß-glucosidase for further ultrasonic stimulation to induce proper stress response. With ultrasound (20 kHz, amplitude at 20%) to irradiate on L. acidophilus BCRC 10695 at stationary phase of growth for 2 min and 24 h of re-incubation, the ß-glucosidase activity was enhanced to 3.91 U/ml, which was 1.82 times of that without ultrasound treatment. Using the ultrasound-treated L. acidophilus BCRC 10695 to ferment soymilk, the fraction of aglycones in total isoflavones in soymilk was effectively increased from 21.8% initially to 97.9% in 24 h. The strategic ultrasound treatment on L. acidophilus BCRC 10695 demonstrated promotion of ß-glucosidase activity, and this methodology had the potential to be applied in the production of functional soymilk by adding probiotics LAB to increase the bioactive isoflavones and nutritional values for human health.

Effect of ultrasound on lactic acid production by Lactobacillus strains in date (Phoenix dactylifera var. Kabkab) syrup.

Date syrup is rich in fermentable sugars and may be used as a substrate for different microbial fermentations, including lactic acid fermentation processes. The beneficial effects of ultrasounds (US) on bioprocesses have been reported for several microorganisms, due to the enhancement of cell growth, as well as improvements in yields and productivities. Therefore, US treatments (30 kHz, 100 W, 10-30 min) were applied to two lactobacilli (Lactobacillus helveticus PTCC 1332 and Lactobacillus acidophilus PTCC 1643), during fermentation using date syrup as substrate. The effects on lactic acid fermentation were evaluated by analyzing cell growth (dry cell weight and viable cell count), substrate consumption (quantification of glucose and fructose), and product formation (quantification of lactic acid) over time. The effects of US were also evaluated on cell membrane permeability. Both lactobacilli were able to grow well on date syrup without the need for addition of further ingredients. The US effects were highly dependent on treatment duration: treatments of 10- and 20-min stimulated lactobacilli growth, while the treatment extension to 30 min negatively affected cell growth. Similarly, the 10- and 20-min treatments increased sugar consumption and lactic acid production, contrarily to the 30-min treatment. All US treatments increased cell membrane permeability, with a more pronounced effect at more extended treatments. The results of this work showed that application of appropriate US treatments could be a useful tool for stimulation of lactic acid production from date syrup, as well as for other fermentative processes that use date syrup as substrate.

Evaluating metabolic response to light exposure in Lactobacillus species via targeted metabolic profiling.

This study reported metabolic profiles of three representative strains from Lactobacillus species, and explored their metabolic response to visible light exposure. We utilized strains from three Lactobacillus species, Lactobacillus acidophilus, Lactobacillus fermentum and Lactobacillus delbrueckii as our model bacteria and applied mass spectrometry base targeted metabolomics to specifically investigate 221 metabolites within multiple metabolic pathways. Similar and diverse metabolome from three tested strains were discovered. Furthermore, all three Lactobacillus strains demonstrated different metabolic profiles in comparison between light expose verse control. In all three strains, 12 metabolites were detected to have significant differences (p-value<0.01) in light exposure culture compared to the control samples (culture grown without light exposure). Principal components analysis using these significantly changed metabolites clearly separated the exposure and control groups in all three studied Lactobacillus strains. Additionally, metabolic pathway impact analysis indicated that several commonly impacted pathways can be observed.

Comparable effects of low-intensity electromagnetic irradiation at the frequency of 51.8 and 53 GHz and antibiotic ceftazidime on Lactobacillus acidophilus growth and survival.

The effects of low-intensity electromagnetic irradiation (EMI) with the frequencies of 51.8 and 53 GHz on Lactobacillus acidophilus growth and survival were revealed. These effects were compared with antibacterial effects of antibiotic ceftazidime. Decrease in bacterial growth rate by EMI was comparable with the inhibitory effect of ceftazidime (minimal inhibitory concentration-16 µM) and no enhanced action was observed with combined effects of EMI and the antibiotic. However, EMI-enhanced antibiotic inhibitory effect on bacterial survival. The kinetics of the bacterial suspension oxidation-reduction potential up to 24 h of the growth was changed by EMI and ceftazidime. The changes were more strongly expressed by combined effects of EMI and antibiotic especially up to 12 h. Moreover, EMI did not change overall energy (glucose)-dependent H(+) efflux across the membrane but it increased N,N'-dicyclohexylcarbodiimide (DCCD)-inhibited H(+) efflux. In contrast, this EMI in combination with ceftazidime decreased DCCD-sensitive H(+) efflux. Low-intensity EMI had inhibitory effect on L. acidophilus bacterial growth and survival. The effect on bacterial survival was more significant in the combination with ceftazidime. The H(+)-translocating F 0 F 1-ATPase, for which DCCD is specific inhibitor, might be a target for EMI and ceftazidime. The revealed bactericide effects on L. acidophilus can be applied in biotechnology, food producing and safety technology.

Viability of Lactobacillus acidophilus in synbiotic guava mousses and its survival under in vitro simulated gastrointestinal conditions.

The effects of refrigeration, freezing and substitution of milk fat by inulin and whey protein concentrate (WPC) on Lactobacillus acidophilus La-5 viability and resistance to gastric and enteric simulated conditions in synbiotic guava mousses effects were investigated. Refrigerated mousses supplemented with WPC presented the best probiotic viability, ranging from 7.77 to 6.24 log cfu/g during 28 days of storage. The highest probiotic populations, above 7.45 log cfu/g, were observed for all frozen mousses during 112 days of storage. Decreased L. acidophilus survival during the in vitro gastrointestinal simulation was observed both for refrigerated and frozen mousses. Nonetheless, for the refrigerated mousses, the addition of inulin enhanced the probiotic survival during the in vitro assays in the first week of storage. L. acidophilus survival in simulated gastrointestinal fluids was also improved through freezing. The frozen storage may be used to provide increased shelf-life for synbiotic guava mousses. Even though the protective effect of inulin and WPC on the probiotic microorganism tested was shown to be more specific for the refrigerated products, the partial replacement of milk fat by these ingredients may also help, as it improves the nutritional value of mousses in both storage conditions.

Electron microscopic analysis of dairy microbes inactivated by ultrasound.

Ultrasonication is a non-thermal method of food preservation that has the advantage of inactivating microbes in food without causing the common side-effects associated with conventional heat treatments, such as nutrient and flavour loss. The aim of this study was to evaluate the use of ultrasound as an alternative to heat pasteurisation and to assess cell damage using transmission electron microscopy (TEM). Three spoilage microbes, previously isolated from pasteurised milk, were used as "test" microbes. Saline solution (SSS) and UHT milk were used as suspension media and were inoculated with exponential growth phase "test" microbes at a microbial concentration of 1 x 10(4) cfu ml(-1). The samples were subjected to power ultrasound (20 kHz, 750 W), at 100%/124 microm wave amplitude for different time intervals. Both Escherichia coli and Saccharomyces cerevisiae were reduced by >99% (for both suspension media) after ultrasonication and Lactobacillus acidophilus was reduced by 72% and 84% in SSS and milk, respectively. Transmission electron microscope micrographs showed that ultrasonication inflicts extensive microbicidal/microbistatic external and internal damage on all three "test" microbes. In E. coli, sonication-induced emulsification caused the formation of unique minute lipopolysaccharide vesicles from the fragmenting cell envelope.

Effect of moderate electric field frequency on growth kinetics and metabolic activity of Lactobacillus acidophilus.

Moderate electric fields (MEF) have been previously shown to alter the metabolic activity of microbial cells; thus, the effect of frequency and electric field would be of considerable interest. We investigated herein the effects of MEF frequency on microbial growth kinetics and bacteriocin (Lacidin A) production of Lactobacillus acidophilus OSU 133 during fermentation. The following fermentation treatments were compared: conventional (for 40 h), MEF (1 V cm(-1), for 40 h), combination of MEF (1 V cm(-1), for the first 5 h) and conventional (for 35 h) at various frequency levels (45, 60, and 90 Hz) all at 30 degrees C, and control (conventional) fermentation at 37 degrees C. MEF treatments with purely sinusoidal waveforms at all frequencies at 30 degrees C produced a shorter lag phase than conventional fermentation. However, no lag phase reduction was found for a 60 Hz waveform that contained high-frequency harmonics. There was, however, a significant increase in the bacteriocin production under early MEF treatment at 60 Hz with high-frequency harmonics. On the basis of these observations, the fermentation process is accelerated by applying pure sinusoidal MEF at the early stage of growth while a significant increase in the bacteriocin production occurs when sinusoidal field at 60 Hz with harmonics is applied at the early stage of the growth.

Effect of moderate electric field on the metabolic activity and growth kinetics of Lactobacillus acidophilus.

Moderate electric fields (MEF), applied across microbial growth media may potentially affect the permeability of cell membranes. We investigated the effects of MEF on bacteriocin (lacidin A) production during fermentation and on microbial growth kinetics of Lactobacillus acidophilus OSU 133. We comparatively investigated the following treatments: conventional, MEF (1 V/cm, 60 Hz, for 40 h), combinations of MEF (1 V/cm, 60 Hz, for the first 5 h) and conventional fermentation (for 35 h), and discrete MEF (1 V/cm, 2 min on and off, for 40 h). In all treatments, except as noted below, temperature was set at 30 degrees C. The two exceptions were control (conventional) and discrete MEF treatment, which were conducted both at 30 and 37 degrees C. MEF treatments at the early stage of fermentation at 30 degrees C showed the maximum bacteriocin activity. Minimum bacteriocin production was observed under conventional fermentation at 37 degrees C. A mathematical model based on Monod growth kinetics was used to predict bacteriocin production and showed results consistent with conventional treatment data. MEF did not have a significant effect on the lag time, maximum specific growth rate, biomass production and pH change under the different experimental conditions at each specific temperature. Based on the observations, bacteriocin activity under the presence of MEF at the early stage of fermentation increased without significant change in the final biomass.

Death of Lactobacillus acidophilus caused by incorporated 3H-thymine during incubation without amino acids.

Of the cells of Lactobacillus acidophilus R-26 incorporated 3H-thymine (specific radioactivity 1.57 Ci/mmol or 3.15 Ci/mmol), their transfer to a medium without essential amino acids resulted in their death. This death may be interpreted in such a way that cell damage caused by disintegration of tritium cannot be effectively repaired under conditions of amino acid deprivation. The experimental eonditions make it possible to explain this death either as a result of inhibition of protein or RNA synthesis or as a result of the absence of amino acids. These possibilities were tested in experiments, in which the synthesis of proteins and RNA was inhibited by specific inhibitors in the presence of amino acids. Under these conditions no death of cells was detected, thus indicating that free amino acids play a role in the repair of radiation damage.