Effect of dilution rate and nutrients addition on the fermentative capability and synthesis of aromatic compounds of two indigenous strains of Saccharomyces cerevisiae in continuous cultures fed with Agave tequilana juice

Knowledge of physiological behavior of indigenous tequila yeast used in fermentation process is still limited. Yeasts have significant impact on the productivity fermentation process as well as the sensorial characteristics of the alcoholic beverage. For these reasons a better knowledge of the physiological and metabolic features of these yeasts is required. The effects of dilution rate, nitrogen and phosphorus source addition and micro-aeration on growth, fermentation and synthesis of volatile compounds of two native Saccharomyces cerevisiae strains, cultured in continuous fed with Agave tequilana juice were studied. For S1 and S2 strains, maximal concentrations of biomass, ethanol, consumed sugars, alcohols and esters were obtained at 0.04 h⁻¹. Those concentrations quickly decreased as D increased. For S. cerevisiae S1 cultures (at D = 0.08 h⁻¹) supplemented with ammonium phosphate (AP) from 1 to 4 g/L, concentrations of residual sugars decreased from 29.42 to 17.60 g/L and ethanol increased from 29.63 to 40.08 g/L, respectively. The S1 culture supplemented with AP was then micro-aerated from 0 to 0.02 vvm, improving all the kinetics parameters: biomass, ethanol and glycerol concentrations increased from 5.66, 40.08 and 3.11 g/L to 8.04, 45.91 and 4.88 g/L; residual sugars decreased from 17.67 g/L to 4.48 g/L; and rates of productions of biomass and ethanol, and consumption of sugars increased from 0.45, 3.21 and 7.33 g/L·h to 0.64, 3.67 and 8.38 g/L·h, respectively. Concentrations of volatile compounds were also influenced by the micro-aeration rate. Ester and alcohol concentrations were higher, in none aerated and in aerated cultures respectively.     

Mass balance and kinetic analysis of anaerobic microbial dechlorination of pentachlorophenol in a continuous flow column

A mass balance and kinetic investigation of anaerobic dechlorination of pentachlorophenol (PCP) was undertaken using an enriched microbial consortium in a laboratory scale continuous flow column, as a model microbial permeable reactive barrier. The chlorine balance showed that 50 µM PCP was largely dechlorinated to phenol with the formation of a small quantity of 3-chlorophenol as an intermediate metabolite (hydraulic retention time 7.6 days), and the chlorine removal efficiency reached 36 µM d^-1. When the initial PCP concentration was increased to 100 µM the chlorine removal efficiency increased 1.5 times. However, the dechlorination activity disappeared after 7.4 pore volumes (58 days), demonstrating the susceptibility of the dechlorination culture to high concentrations of PCP. Lactate released hydrogen as an electron donor during PCP dechlorination, with acetate, propionate, CO₂ and CH₄ as byproducts. The carbon balance showed that some of the organic carbon source (PCP, lactate) in the influent was converted to gas and utilized for biomass growth in addition to organic metabolites. The kinetic study was conducted in a batch culture and yielded 1.99 mg l^-1 biomass growth per unit of chlorine consumption (µM). The Monod equation was well fitted to the specific growth rate of 1.38 d^-1 and a half saturation constant of 0.29 µM. The organic chlorine removal rate in the batch culture was consistent with the results in the flow column, indicating the feasibility of and potential for in situ estimation and prediction through batch culture studies.     

Effect of different fermentation parameters on l-lactic acid production from liquid distillery stillage

Expansion of lactic acid applications, predominantly for the preparation of biodegradable polymers increased the research interest for new, economically favourable production processes. Liquid stillage from bioethanol production can be an inexpensive, valuable source of nutrients for growth of lactic acid bacteria. Utilisation of residual biomass with spent fermentation media as a functional animal feed can greatly influence the process value and its ecological aspect. In this paper, the kinetics of lactic acid and biomass production on liquid stillage by Lactobacillus rhamnosus ATCC 7469 was studied. In addition, the impact of temperature, inoculum concentration, shaking and pH control by addition of CaCO₃ was evaluated. Maximal lactic acid yield of 73.4%, as well as high biomass production (3 × 10⁸ CFU ml⁻¹) were achieved under selected conditions (41 °C, 5% (v/v) of inoculum, 1% (w/v) of CaCO₃, initial pH of 6.5 and shaking rate of 90 rpm). These results were achieved without supplementation of the stillage with nitrogen or mineral sources.     

高速逆流色谱法分离制备大豆异黄酮中的大豆苷和染料木苷

采用高速逆流色谱法分离纯化大豆异黄酮中的大豆苦和染料木昔。溶剂系统为乙酸乙酯一醋酸一水,体积比为5:1:10,上相为固定相,下相为流动相,逆流色谱仪转速为800r/mm,流速为1.5ml/min。所得大豆昔、染料木昔经高效液相色谱分析测定,纯度分别达到98.2%和99.2%(面积归一法)。     

Growth of Carnobacterium divergens M35 and production of Divergicin M35 in snow crab by-product, a natural-grade medium

This work aimed to study the factors influencing the growth of Carnobacterium divergens strain M35 and its ability to produce a new class IIa bacteriocin (divergicin M35) in various synthetic media and in medium supplemented with snow crab hepatopancreas (SCH), a natural-grade by-product of crustacean processing. C. divergens M35 growth and bacteriocin production in SCH-supplemented medium was evaluated at different temperatures in batch fermentations and under controlled pH. C. divergens M35 was shown to grow well in SCH medium at tested temperatures between 4 and 30 °C, except at 37 °C. Maximum divergicin M35 production was obtained after 10 h of growth in SCH medium at 25 and 30 °C with a total activity of 3.7 × 10⁴ AU mL⁻¹. Less growth was observed at 37 °C, for which a total bacteriocin activity of only 256 AU mL⁻¹ was obtained. The production of divergicin M35 was greatly influenced by the medium composition, especially by the type of added carbon source. The best production of divergicin M35 in SCH medium was observed at a controlled pH of 7.0. This study describes the optimal parameters for the growth of C. divergens M35 and production of divergicin M35 and demonstrates the effectiveness of the marine by-product as a medium supplement for this culture.     

野葛细胞固定化培养及异黄酮化合物的生产

以海藻酸钠固定化处理的野葛细胞为试材,研究了固定化细胞培养周期中细胞生长、糖源消耗和异黄酮含量的变化。结果表明,野葛细胞生长曲线呈“S”形,可分为3个主要阶段,即迟滞期(0~8 d)、指数生长期(8~14 d)和稳定增长期(14~18 d)。在培养的第18天达到最大生物量(48.56 g(FW)/L),约为初始接种量的3.04倍。同时,糖源随细胞生长量的增加而不断消耗。在野葛细胞培养周期中,固定化细胞胞内黄酮和培养液黄酮逐渐累积,培养体系中的总黄酮含量与细胞生长量呈同步增长的关系,第16天,总黄酮含量约为初始培养时的17.28倍。试验结果为采用野葛细胞固定化培养的方法进行异黄酮化合物的生产提供了实验依据。     

Effect of fenpropimorph, prochloraz and tebuconazole on growth and production of T-2 and HT-2 toxins by Fusarium langsethiae in oat-based medium

Fusarium langsethiae has been isolated from infected cereals in central and northern Europe where it has been identified in the last decade as the main species involved in the occurrence of high levels of T-2 and HT-2 toxins, mainly in oats. The efficacy of three fungicides (prochloraz, tebuconazole, fenpropimorph) for controlling growth of two strains of F. langsethiae isolated from oats was examined at 0.96 and 0.98 a_w at 15, 20 and 25 °C on oat-based media. The concentrations necessary for 50 and 90% growth inhibition (ED₅₀ and ED₉₀ values) were determined. The effect on the trichothecene type A mycotoxins T-2 and HT-2 was also determined. Without fungicides both strains grew faster at 0.98 than at 0.96 a_w and the influence of temperature on growth rates was 25 > 20 > 15 °C. Prochloraz and tebuconazole were more effective than fenpropimorph against F. langsethiae. Strain, temperature and type of fungicide significantly influenced the ED₅₀ and ED₉₀ values for growth. The concentration ranges under different environmental conditions were: prochloraz (0.03-0.1 and 0.3-1.5), tebuconazole (0.06-0.9 and 1.3-8.2), and fenpropimorph (22-59 and 125-215 mg l⁻¹). Production of T-2 and HT-2 toxins was influenced by temperature, a_w, type of fungicide and dose. Levels of T-2 were usually higher than those of HT-2 under the same conditions. The biosynthesis of T-2 toxin increased after 10 day incubation, but was reduced with decreasing temperature and increasing fungicide dose. At 0.98 a_w T-2 levels increased in cultures containing fenpropimorph while at 0.96 a_w the toxin concentrations increased in response to the other two fungicides. Low doses of prochloraz or tebuconazole enhanced toxin production when compared with untreated cultures for strain 2004-59 at 0.96 a_w and 20-25 °C. HT-2 was hardly detectable in the treatments with prochloraz or tebuconazole at 0.98 a_w. This is the first study on the effect of these anti-fungal compounds on control of growth of F. langsethiae and on production of T-2 and HT-2 toxins in oat-based media.     

Effect of multiple factors on accumulation of nucleosides and bases in Cordyceps militaris

In order to improve the accumulation of valuable metabolites (3′-deoxyadenosine, adenosine, guanosine, cytidine, uridine, adenine and uracil) in Cordyceps militaris, multiple factors such as carbon sources, nitrogen sources, metal ions and fermentation duration were investigated in liquid shake flasks. Using both potato broth and glucose as carbon sources were found to facilitate the production of nucleosides and bases most, and 2% of glucose was the most suitable initial level. Mixture of 0.3% yeast extract with 0.3% peptone was the best selection of nitrogen sources and supplementing with 0.1 mmol/L Mn²⁺ achieved the maximum biomass and biosynthesis of nucleosides and bases among all investigated metal ions. The most suitable harvest time for C. militaris was found to be the 6th day, because the production of 3′-deoxyadenosine, adenosine and uridine began to speed up from the later exponential growth phase until the 6th day. Finally, under optimal culture conditions, the contents of 3′-deoxyadenosine, adenosine, guanosine, cytidine, uridine, adenine and uracil were increased to 0.212 ± 0.014 mg/g, 5.05 ± 0.31 mg/g, 4.03 ± 0.30 mg/g, 0.556 ± 0.029 mg/g, 6.39 ± 0.33 mg/g, 0.208 ± 0.016 mg/g and 0.437 ± 0.027 mg/g, respectively.     

Modeling of pullulan fermentation by using a color variant strain of Aureobasidium pullulans

Pullulan, which is comprised of glucose units, is a simple linear polysaccharide produced by Aureobasidium pullulans. Pullulan has long been used in various applications such as blood plasma substitutes, food additives, adhesive additives, flocculants, and even environmental pollution control agents. Mathematical models of biomass, pullulan, and sucrose profiles during fermentation not only provide information about the kinetic-metabolic nature of pullulan, but also facilitate the control and optimization of pullulan production. In this study, several models were modified and tested in order to describe biomass, pullulan, and sucrose profiles during batch fermentation using a color variant strain of A. pullulans. The results demonstrated that the modified Gompertz model can serve as a universal equation to fit biomass production, pullulan production, and sucrose consumption. Furthermore, validation of this modified Gompertz model indicated that biomass (slope = 1.00, R² = 0.991), pullulan (slope = 1.10, R² = 0.991), and sucrose (slope = 0.96, R² = 0.991) were all predicted accurately.     

Physicochemical factors differentially affect the biomass and bacteriocin production by bovine Enterococcus mundtii CRL1656

Bovine Enterococcus mundtii CRL1656 (Centro de Referencia para Lactobacilos Culture Collection) produces an anti-Listeria and anti-Streptococcus dysgalactiae bacteriocin identified as mundticin CRL1656. The strain and its bacteriocin are candidates to be included in a beneficial product to prevent bovine mastitis as an alternative to antimicrobial agents. To optimize the production of biomass and mundticin CRL1656 by E. mundtii CRL1656, a complete 3 × 2⁴ factorial design was applied. The effect of culture medium, initial pH, inoculum size, incubation temperature, and agitation conditions on biomass and bacteriocin production was evaluated simultaneously. Growth parameters were determined using the modified Gompertz model. A nonlinear model was used to estimate the effects of the variables on growth parameters. Bacteriocin production was analyzed using a linear mixed model. Optimal biomass and mundticin CRL1656 production by E. mundtii CRL1656 were obtained in different conditions. Maximal growth was recorded in autolyzed yeast, peptone, tryptone, Tween 80, and glucose or M17 broths, pH 6.5, 5.0% inoculum, 30°C, with agitation. However, bacteriocin titers were higher in autolyzed yeast, peptone, tryptone, Tween 80, and glucose or de Man-Rogosa-Sharpe (MRS) broths, pH 6.5, 30°C, both with or without agitation. Knowledge of the optimum conditions for growth and bacteriocin production of E. mundtii CRL1656 will allow the obtainment of high levels of biomass and mundticin CRL1656 as bioingredients of potential products to prevent bovine mastitis.     

Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates

Highly reproducible production values of the aminocoumarin antibiotic novobiocin were achieved by cultivation of a heterologous Streptomyces producer strain in commercially available square deepwell plates consisting of 24 wells of 3 ml culture volume each. Between parallel cultivation batches in the deepwell plates, novobiocin accumulation showed standard deviations of 4–9%, compared to 39% in baffled Erlenmeyer flasks. Mycelia used as inoculum could be frozen in the presence of 20% peptone and stored at − 70 °C, allowing repeated cultivations from the same batch of inoculum over extended periods of time. Originally, novobiocin titers in the deepwell plate (5–12 mg l^− 1) were lower than in Erlenmeyer flasks (24 mg l^− 1). Optimization of the inoculation procedure as well as addition of a siloxylated ethylene oxide/propylene oxide copolymer, acting as oxygen carrier, to the production medium increased novobiocin production to 54 mg l^− 1. The additional overexpression of the pathway-specific positive regulator gene novG increased novobiocin production to 163 mg l^− 1. Harvesting the precultures in a defined section of growth phase greatly reduced variability between different batches of inoculum. The use of deepwell plates may considerably reduce the workload and cost of investigations of antibiotic biosynthesis in streptomycetes and other microorganisms due to the high reproducibility and the low requirement for shaker space and culture medium.     

Effect of pre- and postharvest salicylic acid treatment on ethylene production,fungal decay and overall quality of Selva strawberry fruit

The effect of salicylic acid (SA) treatment at different concentrations and growth stages of strawberry (Fragaria ananassa cv. Selva) fruit on postharvest ethylene production, fungal decay and overall quality index was studied. SA at all concentrations effectively reduced fruit ethylene production and fungal decay and retained overall quality. Treatment of plants at vegetative stage and fruit development stage followed by postharvest treatment of fruits with 1 and 2 mmol L⁻¹ was the most effective strategy, whilst with decrease in treatment time the effects of treatment decreased. Single stage treatment strategy of fruits with 2 mmol L⁻¹ SA at postharvest stage was most effective. Postharvest treatment with 4 mmol L⁻¹ SA slightly damaged the fruits and was less effective than 2 mmol L⁻¹ in retaining fruit quality.     

An economic evaluation of freeze-dried kefir starter culture production using whey

An economic study is presented in which industrial-scale production of freeze-dried kefir starter culture is discussed based on results on a laboratory scale. Industrial scale-up was based on a 3-step process using 3 bioreactors of 100, 3,000, and 30,000 L for 300 kg of freeze-dried culture/d of plant capacity. The major cost component of the total investment was the freeze-drying machinery, which consisted of 57% of the total investment. Production cost was reduced from €15.4/kg ($18.5/kg) to ∉2.9/kg ($3.5/kg) when the production capacity was increased from 30 to 900 kg/d. An economic analysis revealed a 3.5-fold increase in production cost compared with the corresponding production cost of the wet product, with an added value of up to ∉10.8 × 10⁹ ($13.0 × 10⁹) within the European Union.     

Effect of supplementing essential fatty acids to pregnant nonlactating Holstein cows and their preweaned calves on calf performance,immune response,and health

The objective was to evaluate the effect of supplementing saturated or unsaturated fatty acids (FA) during late gestation of cows and during the preweaning period of calves on growth, health, and immune responses of calves. During the last 8 wk of pregnancy, Holstein cattle (n = 96) were fed no fat supplement (control), a saturated FA (SFA) supplement enriched in C18:0, or an unsaturated FA supplement enriched in the essential FA linoleic acid. Newborn calves were fed a milk replacer (MR) with either low linoleic acid (LLA; coconut oil) or high linoleic acid (HLA; coconut oil and porcine lard) concentration as the sole feedstuff during the first 30 d. A grain mix with minimal linoleic acid was offered between 31 and 60 d of life. At 30 and 60 d of life, concentrations of linoleic acid in plasma were increased in calves born from dams supplemented with essential FA compared with SFA (44.0 vs. 42.5% of total FA) and in calves consuming HLA compared with LLA MR (46.3 vs. 40.8% of total FA). Total n-3 FA concentration was increased in plasma of calves fed HLA compared with LLA MR (1.44 vs. 1.32%) primarily due to increased α-linolenic acid. Prepartum supplementation with SFA tended to improve dry matter intake (48.8 vs. 46.7 kg) and improved average daily gain (0.50 vs. 0.46 kg/d) by calves without affecting efficiency of gain or circulating concentrations of anabolic metabolites or hormones. Increasing mean intake of linoleic acid from approximately 4.6 to 11.0 g/d during the first 60 d of life increased average daily gain (0.50 vs. 0.45 kg/d) without a change in dry matter intake, thus improving feed efficiency (0.63 vs. 0.59 kg of gain/kg of dry matter intake). Improved weight gain in calves fed HLA MR was accompanied by increased or tendency to increase plasma concentrations of glucose (92.7 vs. 89.9 g/dL) and insulin-like growth factor I (59.5 vs. 53.2 g/dL), increased hematocrit (36.0 vs. 34.4%) and concentration of blood lymphocytes (4.61 vs. 4.21 × 10³/μL), lowered plasma concentrations of acid-soluble protein (78.8 vs. 91.3 mg/L) and blood platelets (736 vs. 822 × 10³/μL), and increased production of IFN-γ by peripheral blood mononuclear cells at 30 d of age (48.1 vs. 25.6 pg/mL), possibly indicating an earlier development of the immune system. Partial replacement of coconut oil with porcine lard in MR improved calf performance and some aspects of immunity.     

Application of a model using the phenomenological approach for prediction of growth and xanthan gum production with sugar cane broth in a batch process

Production of xanthan gum was studied and modelled using unstructured kinetic models composed of three differential equations, which considered the microbial biomass, carbon source, and xanthan concentration. The fermentation process, using Xanthomonas campestris pv. campestris NRRL B-1459, was conducted under controlled conditions with diluted sugar cane broth at different initial sucrose concentrations (15.0, 25.0, and 35.0 g L⁻¹). Unstructured kinetic models proposed in the literature for this system were reviewed and applied. These models were tested against the experimental results, calculating the parameters by nonlinear regression. The kinetic models used in this study provided estimations of microbial growth, substrate consumption, and product formation, and, therefore, these parameters were quantified in the fermentation experiments. Higher yield of xanthan per amount of sucrose (0.58 g g⁻¹) and productivity (0.63 g L⁻¹ h⁻¹) were obtained using initial sucrose concentrations of 25.0 and 35.0 g L⁻¹, respectively. The models were used to predict the kinetic parameters for a medium containing an intermediate and a larger initial sucrose concentration (27.0 and 40.0 g L⁻¹). When tested experimentally, the measured fermentation parameters were in close agreement with the values predicted by the model that presented the best adjustment, demonstrating its validity.     

Effect of light on growth,intracellular and extracellular pigment production by five pigment-producing filamentous fungi in synthetic medium

The competence of the living creatures to sense and respond to light is well known. The effect of darkness and different color light quality on biomass, extracellular and intracellular pigment yield of five potent pigment producers Monascus purpureus, Isaria farinosa, Emericella nidulans, Fusarium verticillioides and Penicillium purpurogenum, with different color shades such as red, pink, reddish brown and yellow, were investigated. Incubation in total darkness increased the biomass, extracellular and intracellular pigment production in all the fungi. Extracellular red pigment produced by M. purpureus resulted maximum in darkness 36.75 ± 2.1 OD and minimum in white unscreened light 5.90 ± 1.1 OD. Similarly, intracellular red pigment produced by M. purpureus resulted maximum in darkness 18.27 ± 0.9 OD/g and minimum in yellow light 8.03 ± 0.6 OD/g of substrate. The maximum biomass production was also noticed in darkness 2.51 g/L and minimum in yellow light 0.5 g/L of dry weight. In contrast, growth of fungi in green and yellow wavelengths resulted in low biomass and pigment yield. It was found that darkness, (red 780–622 nm, blue 492–455 nm) and white light influenced pigment and biomass yield.     

Using a fed-batch culture strategy to enhance rAAV production in the baculovirus/insect cell system

Recombinant adeno-associated virus (rAAV) is one of the most promising vectors for human gene therapy. However, the production systems that are currently available have a limited capacity and cannot provide sufficient quantities of rAAV for preclinical or clinical trials. Many novel methods for improving rAAV production have been developed, but few researchers have focused on the culture process. In this study, we use a fed-batch culture system to enhance rAAV yield in the baculovirus/insect cell system. When the insect cells were co-infected with MOI = 5 of Bac-GFP at a ratio of 1:9:9 (Bac-GFP: Bac-Rep: Bac-VP), the fed-batch culture achieved optimal rAAV yields. In batch culture, the optimal cell density for producing rAAV was found to be 1 × 10⁶ cells/ml, and the highest rAAV yield (1.22 × 10⁸ IVP/ml, 122 IVP/cell) occurred at day 5 post-infection. In the fed-batch culture, rAAV yield reached 2.13 × 10⁸ IVP/ml at day 4 post-infection, and the highest rAAV yield was 2.40 × 10⁸ IVP/ml (240 IVP/cell) at day 5 post-infection. The cost of the batch and fed-batch cultures is similar; however, the rAAV yield was 2.6-fold higher in the fed-batch culture system compared with that in the batch culture system. Therefore, here we demonstrated an economical and efficient strategy for rAAV production.     

Submerged culture conditions for mycelial yield and polysaccharides production by Lyophyllum decastes

The effects of various carbon and nitrogen sources, their concentrations, initial pH and fermentation duration on the production of mycelia in terms of dry weight, exo-polysaccharide (EPS) and inner polysaccharide (IPS) by Lyophyllum decastes, a culinary-medicinal mushroom, were investigated in shake-flask cultures. Lactose, glucose and fructose were the top three best carbon sources for mycelial growth with corresponding yields of 6.73 g/l, 6.36 g/l and 6.10 g/l, respectively. Glucose was the best for production of EPS and IPS with 1.65 g/l and 317 mg/g dry mycelia, respectively. Maltose also performed well for EPS production. Yeast extract was the best nitrogen source for the production of mycelia (7.03 g/l) and IPS (325 mg/g dry mycelia), whereas EPS was improved further by increasing the yeast extract concentration (2.46 g/l at 2%). Similarly, initial pH 7 and 8 were best for polysaccharides production (EPS 1.73 g/l and IPS 320 mg/g) and mycelial growth (7.10 g/l), respectively. Maximum mycelial growth peaked at 15 days of cultivation whereas polysaccharides peaked at 10 days, and then tapered off. A concentration of glucose 3% and yeast extract 1% (mycelial yield and IPS) were found to be a suitable condition for submerged culture.     

Studies on the low-salt Chinese potherb mustard (Brassica juncea, Coss.) pickle. I—The effect of a homofermentative l(+)-lactic acid producer Bacillus coagulans on starter culture in the low-salt Chinese potherb mustard pickle fermentation

A homofermentative l(+)-lactic acid producer Bacillus coagulans B179 has been evaluated as starter cultures for low-salt potherb mustard pickle process. This study examined the impact of inoculation by B. coagulans B179 as starter culture on the spontaneous lab-scale fermentation process of potherb mustard pickle. Three different salt concentrations trials, i.e., 5 g/100 g, 8 g/100 g and 10 g/100 g, respectively, were carried out. As a result, the addition of starter culture of B. coagulans B179 to 5 g/100 g salinity fermentation showed an inhibiting effect upon the population of non-lactic acid bacteria (LAB) and marked increase in population of LAB and production of titratable acidity but did not significantly influence the pickle end products quantity of acid produced by fermentation as compared with a treatment without inoculation. However, in the higher salt concentrations, i.e., 8 g/100 g and 10 g/100 g NaCl trials, inoculation had no impact on the fermentation of the pickle. From the standpoint of fermentation, as a functional starter culture, inoculated B. coagulans B179 can effectively improve desired native LAB growth and inhibit the growth of undesired fungi, which will thereby allow it to shorten fermentation period and enhance the pickle quality in the low-salt potherb mustard pickle production.