Multilocus sequence typing of L. bulgaricus and S. thermophilus strains from Turkish traditional yoghurts and characterisation of their techno-functional roles.

In this study, Streptococcus thermophilus and Lactobacillus bulgaricus strains from traditional Turkish yoghurts were isolated, identified by 16S rRNA sequencing and genotypically 14 S. thermophilus and 6 L. bulgaricus strains were obtained as distinct strains by MLST analysis. Lactic acid production levels of the L. bulgaricus strains were higher than S. thermophilus strains. HPLC analysis showed that EPS monosaccharide composition of the strains mainly consisted of glucose and galactose. In general, all strains were found to be susceptible for antibiotics, except some strains were resistance to gentamicin and kanamycin. Apart from two strains of S. thermophilus, all strains displayed strong auto-aggregation level greater than 95% at 24 h incubation. S. thermophilus strains showed higher cell surface hydrophobicity than L. bulgaricus strains. This study demonstrated the isolation, identification, genotypic discrimination and techno-functional features of wild type yoghurt starter cultures which can potentially find place in industrial applications. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01366-2.

Isolation and characterization of yogurt starter cultures from traditional yogurts and growth kinetics of selected cultures under lab-scale fermentation.

The development of new starter cultures is a crucial task for the food industry to meet technological requirements and traditional products are important reservoirs for new starter cultures. In this respect, this study aimed to isolate, identify, and determine the technological characteristics of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains originated from traditional yogurt samples. Genotypic discrimination of 200 isolates revealed the presence of distinct 19 S. thermophilus and 11 Lb. delbrueckii subsp. bulgaricus strains as potential starter cultures. Strain-specific properties determined the acidification capacity of the yogurt starter cultures and a higher acidification capacity was observed for S. thermophilus strains compared to Lb. delbrueckii subsp. bulgaricus strains. Proteolytic activity was found between 0.012-0.172 and 0.078-0.406 for S. thermophilus and Lb. delbrueckii subsp. bulgaricus strains, respectively. 4 of S. thermophilus and 3 of Lb. delbrueckii subsp. bulgaricus strains were found resistant to all tested bacteriophages. The antibiotic susceptibility tests of the isolates revealed that a very low antibiotic resistance was observed for the yogurt starter cultures. Finally, the growth kinetics of selected strains were determined and the maximum specific growth rate of selected S. thermophilus and Lb. delbrueckii subsp. bulgaricus was calculated as 0.527 h-1 and 0.589 h-1, respectively.

Antifungal Activities of Different Essential Oils and Their Electrospun Nanofibers against Aspergillus and Penicillium Species Isolated from Bread.

Mold growth, especially Aspergillus spp. and Penicillium spp., deteriorates the quality of bakery products. Essential oils (EOs) have been categorized as good natural antimicrobials. Hereby, this study aimed to evaluate the antifungal activity of six EOs, ginger, cumin, cinnamon, black pepper, origanum, and clove, and their volatile compounds against fungal strains isolated from bread: Penicillium carneum DDS4, Aspergillus flavus DDS6, and Aspergillus niger DDS7 by disc diffusion and disc volatilization methods, respectively. Among EOs, cumin, cinnamon, origanum, and clove were found to be effective against fungal strains, and their minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined. The observed lowest MIC value of EOs was obtained at 1000 µg/mL concentration, and the lowest MFC value was obtained from the results of clove at a concentration of 1000 µg/mL. Based on the MIC and MFC values, clove and cinnamon EOs were found to be more effective at lower concentrations. Electrospun nanofiber films of clove and cinnamon were produced with 6% poly(vinyl alcohol) (PVA), 2% ß-cyclodextrin (ß-CD), and 2% EO to overcome the unfavorable sensory impact of EOs on food products. The inhibitory activity of cinnamon EO film (2.64-2.51 log(CFU/mg)) was considerably lower than clove EO film (3.18-3.24 log(CFU/mg)) against P. carneum DDS4 and A. niger DDS7. Furthermore, these nanofiber films prevented fungal growth on bread samples visibly and were shown to be an alternative application for active food packaging.

High level production of itaconic acid at low pH by Ustilago maydis with fed-batch fermentation.

The metabolically engineered plant pathogen Ustilago maydis MB215 Δcyp3 Petefria1 has been cultivated to produce more than 80 g/L itaconate in 16 L scale pH and temperature controlled fermentation, in fed-batch mode with two successive feedings. The effect of pH as well as successive rounds of feeding has been quantified via elemental balances. Volumetric itaconic acid productivity gradually decreased with successive glucose feedings with increasing itaconic titers, with nearly constant product yield. Extracellular pH was decreased from 6 down to 3.5 and the fermentation was characterized in specific uptake, production, and growth rates. Notable is that the biomass composition changes significantly from growth phase to itaconic acid production phase, carbon content increases from 42% to around 62%. Despite the gradual decrease in itaconic acid levels with decreasing pH (nearly 50% decrease in itaconic acid at pH 3.5, compared to pH 6), significant itaconate production is still observed at pH 4 (around 63 g/L). Biomass yield remained nearly constant until pH 4. Taken together, these results strongly illustrate the potential of engineered Ustilago maydis in itaconate production at commercial levels.

Long term performance of a pilot scale anaerobic membrane bioreactor treating beet molasses based industrial wastewater.

Baker's yeast industries (BYI) generate highly polluted effluents, especially vinasse from yeast separators, with very high chemical oxygen demand (COD), nitrogen, sulphate and salts, mainly potassium and calcium. Anaerobic treatment is the most commonly applied method for treating BYI wastewaters. However, it is quite challenging to obtain a high performance due to the difficulties in biomass retention. Moreover, it does not provide compliance with COD and color discharge limits when used as a sole treatment process. In this context, a pilot scale anaerobic membrane bioreactor, which provides excellent biomass retention, was operated to investigate its treatment performance for vinasse from a BYI. The reactor achieved a COD removal between 48% and 92% up to a volumetric load of 10 kg COD m3 d-1. A specific methane production of 0.37 m3 CH4 kg-1 CODremoved was observed in the study. On the other hand, passage of inert organic compounds through membrane deteriorated permeate quality and treatment efficiency. High alkalinity and pH led to the accumulation of calcium precipitates, which reduced volatile solids fraction of sludge and biomass activity in the reactor. The present study showed the operational challenges and potential drawbacks of AnMBR systems for BYI wastewater treatment. The experience gained in the pilot system can be utilized in the design and operation of full scale AnMBRs for high strength industrial effluents.

Removal of dimethyl sulphide via a bio-scrubber under anoxic conditions.

The removal performance of dimethyl sulphide (DMS) by anoxic laboratory-scale bio-scrubber was studied under different operation conditions for 315 days. DMS removal in bio-scrubber system was performed by controlling and changing the operation parameters, including inlet concentration, empty bed residence time (EBRT) and spraying density (SD) of irrigation. Best conditions in the system were achieved for SD of 0.18 m3/m2 h within EBRT of 40 s at an inlet gas concentration of 150 mg/m3 in which 93% of waste gas stream was removed in the bio-scrubber column and bio-degradation in the bio-reactor tank led to 89% of DMS removal from the transferred bio-reactor, while 91.5% of input chemical oxygen demand (COD) was successfully removed. The use of closer values of the average experimental yield to the theoretical value (YNO3/NO3 -) of 0.74 led to the production of elemental sulphur (S°) and other sulphur forms rather than sulphate (SO42-) , which was also was recognized as a pale-yellow coloured substance of S° that appeared within the biomass.

Removal of ethanethiol using a biotrickling filter with nitrate as an electron acceptor.

Many studies have discussed the biotreatment of ethanethiol (ET) under aerobic conditions. However, O2 free conditions offer bio-conversion of ET gas into elemental sulphur and/or sulphate using [Formula: see text] as electron acceptor, and this has been not studied. In this study, an anoxic biotrickling filter was tested in lab-scale conditions with ET/[Formula: see text] ratio 0.74 and 0.34 mole/mole to remove malodorous ET waste gas. The study examined the effect of three operational parameters: ET inlet concentrations (150, 300, 800, and 1500 mg/m3), trickling velocities (0.12, 0.18, 0.24, 0.3, and 0.45 m/h), and empty bed residence times (30, 60, 90, and 120 s). It found that the effect of trickling velocity on removal efficiency depended on inlet concentrations; 0.24 m/h trickling velocity resulted in efficient ET removal (higher than 90.8% for 150 mg/m3 of inlet concentration) while 0.45 m/h trickling velocity could only achieve a removal of 80.6% for 1500 mg/m3 of inlet concentration at fixed EBRT 60 s. Increasing the EBRT up to 60 s was adequate to achieve removal efficiency, i.e. 92 and 80% for ET inlet concentrations 150 and 1500 mg/m3 respectively, and the maximum elimination capacity was 75.18 g/m3/h at 0.45 m/h. Overall, the anoxic conditions enhanced the low oxidation rates of ET in an anoxic biotrickling filter despite mass transfer limitations and poor solubility of ET.

Metabolic engineering with ATP-citrate lyase and nitrogen source supplementation improves itaconic acid production in Aspergillus niger.

BACKGROUND: Bio-based production of organic acids promises to be an attractive alternative for the chemicals industry to substitute petrochemicals as building-block chemicals. In recent years, itaconic acid (IA, methylenesuccinic acid) has been established as a sustainable building-block chemical for the manufacture of various products such as synthetic resins, coatings, and biofuels. The natural IA producer Aspergillus terreus is currently used for industrial IA production; however, the filamentous fungus Aspergillus niger has been suggested to be a more suitable host for this purpose. In our previous report, we communicated the overexpression of a putative cytosolic citrate synthase citB in an A. niger strain carrying the full IA biosynthesis gene cluster from A. terreus, which resulted in the highest final titer reported for A. niger (26.2 g/L IA). In this research, we have attempted to improve this pathway by increasing the cytosolic acetyl-CoA pool. Additionally, we have also performed fermentation optimization by varying the nitrogen source and concentration. RESULTS: To increase the cytosolic acetyl-CoA pool, we have overexpressed genes acl1 and acl2 that together encode for ATP-citrate lyase (ACL). Metabolic engineering of ACL resulted in improved IA production through an apparent increase in glycolytic flux. Strains that overexpress acl12 show an increased yield, titer and productivity in comparison with parental strain CitB#99. Furthermore, IA fermentation conditions were improved by nitrogen supplementation, which resulted in alkalization of the medium and thereby reducing IA-induced weak-acid stress. In turn, the alkalizing effect of nitrogen supplementation enabled an elongated idiophase and allowed final titers up to 42.7 g/L to be reached at a productivity of 0.18 g/L/h and yield of 0.26 g/g in 10-L bioreactors. CONCLUSION: Ultimately, this study shows that metabolic engineering of ACL in our rewired IA biosynthesis pathway leads to improved IA production in A. niger due to an increase in glycolytic flux. Furthermore, IA fermentation conditions were improved by nitrogen supplementation that alleviates IA induced weak-acid stress and extends the idiophase.

Ethanethiol gas removal in an anoxic bio-scrubber.

The performance of ethanethiol removal in an anoxic lab-scale bio-scrubber was investigated under different operating parameters and conditions for 300 days. The removal efficiency (RE) of ethanethiol was examined as a function of inlet concentration, empty bed residence time (EBRT) and spray density of irrigation. The results showed the best operation conditions and operation characteristics of the bio-scrubber for this study were at an inlet concentration of 150 mg/m3, a spray density of 0.23 m3/m2 h and an EBRT of 90 s. An average RE of 91% and elimination capacity (EC) of 24.74 g/m3 h was found for all inlet ethanethiol concentrations. Variations in spray density higher than 0.23 m3/m2 h had no effect on ethanethiol RE at different ethanethiol concentrations. The average experimental yield values were closer to the YET/NO3- theoretical value of 0.74 when the main product was elemental sulphur (So). This indicates that So and other forms of sulphur were formed rather than sulphate (SO42-) as the end product. Furthermore, growth kinetics for bio-degradation were evaluated in batch culture experiments using the Monod model, and bio-kinetic parameters of µmax, Ks, Yxs and qmax were obtained as 0.14 1/h, 1.17 mg/L, 0.52 gx/gs and 0.26 gs/gx h, respectively.

Er:Yag Laser Treatment of Simple Snorers in an Outpatient Setting.

OBJECTIVE: Snoring occurs as a result of soft tissue vibration caused by a partial upper airway collapse during sleep. This study evaluated the effectiveness and potential adverse side effects associated with the use of a nonsurgical, erbium-doped yttrium aluminum garnet (Er:YAG) laser treatment for patients with snoring conditions. MATERIALS AND METHODS: In total, 33 patients with different degrees of snoring were analyzed retrospectively. All patients received three NightLase™ Er:YAG laser treatments. Results were measured using a follow-up questionnaire and then statistically analyzed. Any effects that occurred during the first year after treatment (i.e., short-term effects) were followed up with interviews. RESULTS: Laser treatment effectively reduced patients' snoring and achieved a 65% satisfaction rate after three treatments. The greatest improvement and satisfaction were experienced by patients aged ≥50 years. Patients reported additional benefits from this treatment including easier breathing, higher alertness, and increased focus. CONCLUSION: Nonsurgical Er:YAG laser treatment is an effective and minimally invasive procedure to reduce patient snoring and other sleep-disordered breathing symptoms. Patients reported minimal disadvantages including minor discomfort and a low risk of side effects.

Sulfide removal from industrial wastewaters by lithotrophic denitrification using nitrate as an electron acceptor.

Sulfide is present in wastewaters as well as in biogas and can be removed by several physicochemical and biotechnological processes. Nitrate is a potential electron acceptor, readily available in most wastewater treatment plants and it can replace oxygen under anoxic conditions. A lab-scale reactor was operated for treatment of sulfide containing wastewater with nitrate as an electron acceptor and is used to evaluate the effects of volumetric loading rates, hydraulic retention time (HRT) and substrate concentrations on the performance of the lithotrophic denitrification process for treating industrial fermentation wastewaters. Sulfide is removed more than 90% at the loading rates between 0.055 and 2.004 kg S(-2)/m(3) d, when the influent sulfide concentration is kept around 0.163 kg/m(3) and the HRT decreased from 86.4 to 2 h. Nitrogen removal differed between 23 and 99% with different influent NO(3)(-)-N concentration and loading rates of NO(3)(-)/S(-2) ratio. The stoichiometry of sulfide oxidation with nitrate is calculated assuming different end-products based on thermodynamic approach and compared with experimental yield values. The calculated maximum volumetric and specific sulfide oxidation rates reached 0.076 kg S(-2)/m(3) h and 0.11 kg S(-2)/kg VSS h, respectively. The results are obtained at industrially relevant conditions and can be easily adapted to either biogas cleaning process or to sulfide containing effluent streams.

On-line evolutionary optimization of an industrial fed-batch yeast fermentation process.

This paper presents two genetic algorithms based on optimization methods to maximize biomass concentration, and to minimize ethanol formation. The objective function is maximized according to the values of feed flow rate, using genetic search approaches. Five case studies were carried out for different initial conditions, which strongly influence the optimal profiles of feed flow rate for the fermentation process. The ethanol and glucose disturbance effects were examined to stress the effectiveness of proposed approaches. The proposed genetic approaches were implemented for an industrial scale baker's yeast fermentor which produces Saccharomyces cerevisiae known as baker's yeast. The results show that optimal feed flow rate was obtained in a satisfactory and successful way for fed-batch fermentation process.

Removal of ammonia as struvite from anaerobic digester effluents and recycling of magnesium and phosphate.

A second order kinetic model was developed to predict the rate and extent of NH(4)(+) removal as struvite from anaerobic digester effluents. Alternative to this, NH(4)(+) can be recovered from struvite and the remaining Mg(2+) and PO(4)(3-) can be recycled back to the wastewater to fix more NH(4)(+). The NH(4)(+) solution was retained and the remaining Mg(2+) and PO(4)(3-) were returned back to be mixed with wastewater. In a five-step process, NH(4)(+) recovery was initially 92% and progressively decreased to 77% in the fifth stage, due to loss of Mg(2+) and PO(4)(3-) at each step in the supernatant. Finally, economic analysis of recycling nutrients was performed and compared to the one step process. The cost of NH(4)(+) recovery was calculated as $0.36/kgNH(4)-N which is lower than $7.7/kgNH(4)-N the cost of one step process without considering the market value of struvite obtained in one step process.

Nonlinear predictive control of a drying process using genetic algorithms.

A nonlinear predictive control technique is developed to determine the optimal drying profile for a drying process. A complete nonlinear model of the baker's yeast drying process is used for predicting the future control actions. To minimize the difference between the model predictions and the desired trajectory throughout finite horizon, an objective function is described. The optimization problem is solved using a genetic algorithm due to the successful overconventional optimization techniques in the applications of the complex optimization problems. The control scheme comprises a drying process, a nonlinear prediction model, an optimizer, and a genetic search block. The nonlinear predictive control method proposed in this paper is applied to the baker's yeast drying process. The results show significant enhancement of the manufacturing quality, considerable decrease of the energy consumption and drying time, obtained by the proposed nonlinear predictive control.

Indirect spinal canal decompression of vertebral burst fracture in calf model.

INTRODUCTION: An experimental study of experimental burst fractures in bovine spinal specimens was conducted to analyze the effects of transpedicular short-segment posterior fixation followed by reduction on indirect spinal canal decompression. MATERIALS AND METHODS: For this purpose, experimental burst fractures were created in 11 bovine specimens with a hydraulic materials-testing machine. The specimens were evaluated with plain radiographs and CT scans before reduction. Thereafter, they were instrumented with titanium transpedicular screws and rods (short-segment posterior fixation); and reduction was achieved which included distraction and kyphosis correction maneuvers. RESULTS: Each spinal specimen was evaluated with plain radiographs and CT scans after reduction by applying distraction and kyphosis correction maneuvers. Plain radiographic analysis showed that the kyphosis angle and segmental height values improved. Furthermore, CT scans revealed that the spinal canal diameter values improved compared with those before reduction. The differences between before and after reduction in kyphosis angle, segmental height, anterior body compression, and percentage of retropulsion were statistically significant. CONCLUSION: Short-segment posterior fixation followed by indirect spinal canal decompression led to an improvement over spinal canal retropulsion in experimental burst fractures. Furthermore, the kyphosis angle and segmental height values improved following the reduction compared with those before reduction.