Lag Phase Of Growth Research Articles

Development and Validation of Predictive Model for Salmonella Growth in Unpasteurized Liquid Eggs.

Liquid egg products can be contaminated with Salmonella spp. during processing. A predictive model for the growth of Salmonella spp. in unpasteurized liquid eggs was developed and validated. Liquid whole egg, liquid yolk, and liquid egg white samples were prepared and inoculated with Salmonella mixture (approximately 3 Log CFU/mL) containing five serovars (S. Bareilly, S. Richmond, S. Typhimurium monophasic, S. Enteritidis, and S. Gallinarum). Salmonella growth data at isothermal temperatures (5, 10, 15, 20, 25, 30, 35, and 40°C) was collected by 960 h. The population of Salmonella in liquid whole egg and egg yolk increased at above 10°C, while Salmonella in egg white did not proliferate at all temperature. These results demonstrate that there is a difference in the growth of Salmonella depending on the types of liquid eggs (egg yolk, egg white, liquid whole egg) and storage temperature. To fit the growth data of Salmonella in liquid whole egg and egg yolk, Baranyi model was used as the primary model and the maximum growth rate and lag phase duration for each temperature were determined. A secondary model was developed with maximum growth rate as a function of temperature. The model performance measures, bias factor (Bf, 0.96-0.99) and r2 (0.96-0.99) indicated good fit for both primary and secondary models. In conclusion, it is thought that the growth model can be used usefully to predict Salmonella spp. growth in various types of unpasteurized liquid eggs when those are exposed to various temperature and time conditions during the processing.

Interaction of Mannose-Binding Lectin With Lipopolysaccharide Outer Core Region and Its Biological Consequences.

Lipopolysaccharide (LPS, endotoxin), the main surface antigen and virulence factor of Gram-negative bacteria, is composed of lipid A, core oligosaccharide, and O-specific polysaccharide (O-PS) regions. Each LPS region is capable of complement activation. We have demonstrated that LPS of Hafnia alvei, an opportunistic human pathogen, reacts strongly with human and murine mannose-binding lectins (MBLs). Moreover, MBL–LPS interactions were detected for the majority of other Gram-negative species investigated. H. alvei was used as a model pathogen to investigate the biological consequences of these interactions. The core oligosaccharide region of H. alvei LPS was identified as the main target for human and murine MBL, especially l-glycero-d-manno-heptose (Hep) and N-acetyl-d-glucosamine (GlcNAc) residues within the outer core region. MBL-binding motifs of LPS are accessible to MBL on the surface of bacterial cells and LPS aggregates. Generally, the accessibility of outer core structures for interaction with MBL is highest during the lag phase of bacterial growth. The LPS core oligosaccharide–MBL interactions led to complement activation and also induced an anaphylactoid shock in mice. Unlike Klebsiella pneumoniae O3 LPS, robust lectin pathway activation of H. alvei LPS in vivo was mainly the result of outer core recognition by MBL; involvement of the O-PS is not necessary for anaphylactoid shock induction. Our results contribute to a better understanding of MBL–LPS interaction and may support development of therapeutic strategies against sepsis based on complement inhibition.

Automated Cell Treatment for Competence and Transformation of Escherichia coli in a High-Throughput Quasi-Turbidostat Using Microtiter Plates.

Metabolic engineering and genome editing strategies often lead to large strain libraries of a bacterial host. Nevertheless, the generation of competent cells is the basis for transformation and subsequent screening of these strains. While preparation of competent cells is a standard procedure in flask cultivations, parallelization becomes a challenging task when working with larger libraries and liquid handling stations as transformation efficiency depends on a distinct physiological state of the cells. We present a robust method for the preparation of competent cells and their transformation. The strength of the method is that all cells on the plate can be maintained at a high growth rate until all cultures have reached a defined cell density regardless of growth rate and lag phase variabilities. This allows sufficient transformation in automated high throughput facilities and solves important scheduling issues in wet-lab library screenings. We address the problem of different growth rates, lag phases, and initial cell densities inspired by the characteristics of continuous cultures. The method functions on a fully automated liquid handling platform including all steps from the inoculation of the liquid cultures to plating and incubation on agar plates. The key advantage of the developed method is that it enables cell harvest in 96 well plates at a predefined time by keeping fast growing cells in the exponential phase as in turbidostat cultivations. This is done by a periodic monitoring of cell growth and a controlled dilution specific for each well. With the described methodology, we were able to transform different strains in parallel. The transformants produced can be picked and used in further automated screening experiments. This method offers the possibility to transform any combination of strain- and plasmid library in an automated high-throughput system, overcoming an important bottleneck in the high-throughput screening and the overall chain of bioprocess development.

Growth patterns for three generations of an intercross between red junglefowl and chickens selected for low body weight.

Growth is a complex and dynamic process that may be measured at a specific point or over a period of time. Compared was the growth of male and female chickens over a three-generation period. Involved were red junglefowl (RJF; Gallus gallus), a line of White Plymouth Rock chickens (LWS; Gallus gallus domesticus) selected for low body weight, and their reciprocal F1 and F2 crosses. In both sexes, Gompertz's description of growth showed that RJF had significantly lower asymptotes, earlier inflection points, and faster growth rates than LWS. Heterosis for these measures was positive for asymptote and negative for growth rate and inflection point. The RJF commenced egg production at a significantly younger age and lower body weight than LWS. Although F1 and F2 reciprocal crosses were similar for body weight and for age at first egg, the F1 reciprocal crosses began lay at significantly younger ages than the F2 crosses and parental lines. When viewed on a physiological basis where age and body weight were simultaneously standardized, both parental lines and reciprocal F1 and F2 crosses had differing rapid and lag growth phases. Overall, sexual dimorphism increased in all populations from hatch to sexual maturity. The LWS males had a longer growth period consistent with their female counterparts who became sexually mature at older ages. Comprehensively, these results indicate additive and nonadditive genetic variation for distinct growth patterns and changes in resource allocation strategies over time.

Kinetic models for the effect of temperature on flavonoid production in liquid submerged fermentation by Phellinus baumii.

Kinetic models and temperature control strategy were established to reflect the effect of temperature (22°C-30°C) on flavonoid production of Phellinus baumii (P. baumii) in 6-L fermentor. A modified Logistic equation, Hinshelwood model, and Luedeking Piret equation were used to describe mycelial growth and product formation. The influence of temperature on the estimated kinetic parameters was further studied by regression analysis. Based on kinetic parameters analysis, the new temperature control strategy was proposed. Briefly, at 0-43H, decreasing temperature (30°C-28°C) can shorten the lag phase of mycelial growth, and at 43-90H, fermentation temperature was reduced gradually from 28°C to 24°C to keep high flavonoid productivity. At the fermentation anaphase (90-161H), temperature was controlled at 24°C to relieve inhibition of flavonoid and maintain constant production capacity of flavonoid. As a result, the maximum flavonoid yield was reached 4.21mg/100mg cell dry weight by temperature control strategy, which was 70.45% higher than that at a constant temperature of 26°C. Additionally, the establishment of kinetic models based on fermentation temperature, which presented here may provide a scientific basis for further large scales flavonoid production of P. baumii in submerged fermentation.

Reduction of infection risk mediated by co-culturing Vibrio parahaemolyticus and Listeria monocytogenes in refrigerated cooked shrimp.

Vibrio parahaemolyticus and Listeria monocytogenes are seafood pathogens of public health significance, and predictive models are effective tools for quantitative microbial risk assessment of these pathogens. However, most current predictive models are based on growth of single strains in broth cultures, and interactions of two or more bacteria in a food matrix can skew the outcomes of the predictions. Therefore, the impact of V. parahaemolyticus and L. monocytogenes when co-cultured and in monoculture on cooked shrimp in cold storage was investigated. The results indicated that L. monocytogenes co-cultured with V. parahaemolyticus exhibited reduced growth and longer lag phase at 4 °C and 10 °C. V. parahaemolyticus exhibited similar behavior when co-cultured with L. monocytogenes at 4 °C (death rate K=-0.67 log10 CFU g-1 day . The death rate K at 10 °C when V. parahaemolyticus co-cultured with L. monocytogenes was -1.62 log10 CFU g-1 day-1 . There was no significant reduction of growth in monoculture experiments. This study has revealed that interaction of V. parahaemolyticus and L. monocytogenes should be considered when quantifying risks posed by these pathogens during consumption of seafood products. © 2018 Society of Chemical Industry.

Listeria monocytogenes Growth Kinetics in Milkshakes Made from Naturally and Artificially Contaminated Ice Cream.

This study assessed the growth of Listeria monocytogenes in milkshakes made using the process-contaminated ice cream associated with a listeriosis outbreak in comparison to milkshakes made with artificially contaminated ice cream. For all temperatures, growth kinetics including growth rates, lag phases, maximum populations, and population increases were determined for the naturally and artificially derived contaminants at 5, 10, 15, and 25°C storage for 144 h. The artificially inoculated L. monocytogenes presented lower growth rates and shorter lag phases than the naturally contaminated populations at all temperatures except for 5°C, where the reverse was observed. At 25°C, lag phases of the naturally and artificially contaminated L. monocytogenes were 11.6 and 7.8 h, respectively. The highest increase in population was observed for the artificially inoculated pathogen at 15°C after 96 h (6.16 log CFU/mL) of storage. Growth models for both contamination states in milkshakes were determined. In addition, this study evaluated the antimicrobial effectiveness of flavoring agents, including strawberry, chocolate and mint, on the growth of the pathogen in milkshakes during 10°C storage. All flavor additions resulted in decreased growth rates of L. monocytogenes for both contamination states. The addition of chocolate and mint flavoring also resulted in significantly longer lag phases for both contamination states. This study provides insight into the differences in growth between naturally and artificially contaminated L. monocytogenes in a food product.

Lipids, pH, and Their Interaction Affect the Inhibitory Effects of Carvacrol against Salmonella Typhimurium PT4 and Escherichia coli O157:H7.

Although carvacrol (CAR) is considered an alternative antimicrobial for use in food, few is known about the influence of food-related parameters on its inhibitory effects against pathogens. This study assessed the influence of different amounts of proteins, using beef extract (BE) as a protein-rich source, lipids (LIP), using sunflower oil as a LIP-rich source, and pH values or their interaction on the inhibitory effects of CAR against Salmonella Typhimurium PT4 (ST) and Escherichia coli O157:H7 (EC). The specific maximum growth rate (μmax) and lag phase duration (λ) of the test pathogens when exposed to CAR in media with different amounts of BE (4, 6, and 8 g/100 mL), LIP (3.75, 5, and 6.25 mL/100 mL), and pH values (5, 5.5, and 6) were determined. The viable counts of the tested pathogens in media that promoted the highest and lowest μmax in the presence of CAR were monitored during 24 h. The lowest μmax of ST and EC exposed to 2.4 μL/mL (-1.29 and -0.82 log CFU/mL/h, respectively) or 4.8 μL/mL CAR (-1.44 and -2.17 log CFU/mL/h, respectively) were observed in media with the highest LIP amount (6.25 mL/100 mL) and pH value (pH 6). For both SE and EC, the longest λ (> 2 h) was verified in media where these pathogens showed the lowest μmax. These data indicate that the concomitant increase in LIP amounts and pH values affected positively the CAR inhibitory effects against the target pathogens. CAR (2.4 or 4.8 μL/mL) failed to inhibit the increase in ST and EC counts in media where the highest μmax values were previously observed. On the contrary, CAR inhibited the increase of ST counts (final counts 5 log CFU/mL) and decreased the EC counts (final counts 3.5 log CFU/mL) in media where the lowest μmax values were observed. These results show that the inhibitory effects of CAR on ST and EC in food matrices could be affected as a function of the interaction of LIP amounts and pH values.

Mathematical modelling of temperature effect on growth kinetics of Pseudomonas spp. on sliced mushroom (Agaricus bisporus)

The growth data of Pseudomonas spp. on sliced mushrooms (Agaricus bisporus) stored between 4 and 28°C were obtained and fitted to three different primary models, known as the modified Gompertz, logistic and Baranyi models. The goodness of fit of these models was compared by considering the mean squared error (MSE) and the coefficient of determination for nonlinear regression (pseudo-R2). The Baranyi model yielded the lowest MSE and highest pseudo-R2 values. Therefore, the Baranyi model was selected as the best primary model. Maximum specific growth rate (rmax) and lag phase duration (λ) obtained from the Baranyi model were fitted to secondary models namely, the Ratkowsky and Arrhenius models. High pseudo-R2 and low MSE values indicated that the Arrhenius model has a high goodness of fit to determine the effect of temperature on rmax. Observed number of Pseudomonas spp. on sliced mushrooms from independent experiments was compared with the predicted number of Pseudomonas spp. with the models used by considering the Bf and Af values. The Bf and Af values were found to be 0.974 and 1.036, respectively. The correlation between the observed and predicted number of Pseudomonas spp. was high. Mushroom spoilage was simulated as a function of temperature with the models used. The models used for Pseudomonas spp. growth can provide a fast and cost-effective alternative to traditional microbiological techniques to determine the effect of storage temperature on product shelf-life. The models can be used to evaluate the growth behaviour of Pseudomonas spp. on sliced mushroom, set limits for the quantitative detection of the microbial spoilage and assess product shelf-life.

Nisin in Combination with Cinnamaldehyde and EDTA to Control Growth of Escherichia coli Strains of Swine Origin.

Post-weaning diarrhoea (PWD) due to enterotoxigenic Escherichia coli (ETEC) is an economically important disease in pig production worldwide. Although antibiotics have contributed significantly to mitigate the economic losses caused by PWD, there is major concern over the increased incidence of antimicrobial resistance among bacteria isolated from pigs. Consequently, suitable alternatives that are safe and effective are urgently required. Many naturally occurring compounds, including the antimicrobial peptide nisin and a number of plant essential oils, have been widely studied and are reported to be effective as antimicrobial agents against pathogenic microorganisms. Here, we evaluate the potential of nisin in combination with the essential oil cinnamaldehyde and ethylenediaminetetraacetic acid (EDTA) to control the growth of E. coli strains of swine origin including two characterized as ETEC. The results reveal that the use of nisin (10 μM) with low concentrations of trans-cinnamaldehyde (125 μg/mL) and EDTA (0.25–2%) resulted in extended lag phases of growth compared to when either antimicrobial is used alone. Further analysis through kill curves revealed that an approximate 1-log reduction in E. coli cell counts was observed against the majority of targets tested following 3 h incubation. These results highlight the potential benefits of combining the natural antimicrobial nisin with trans-cinnamaldehyde and EDTA as a new approach for the inhibition of E. coli strains of swine origin.

Biotransformation and detoxification of selenite by microbial biogenesis of selenium-sulfur nanoparticles

This study combines the interaction between the toxic oxyanions selenite and selenate and the plant growth promoting bacterium Azospirillum brasilense with a comprehensive characterization of the formed selenium particles. As selenium is an essential trace element, but also toxic in high concentrations, its state of occurrence in nature is of major concern. Growth of the bacterium was affected by selenite (1–5mM) only, observable as a prolonged growth lag-phase of 3days. Subsequently, selenite reduction occurred under aerobic conditions resulting in extracellularly formed insoluble Se0 particles. Complementary studies by microscopic and spectroscopic techniques revealed the particles to be homogeneous and stable Se8-nSn structured spheres with an average size of 400nm and highly negative surface charge of −18mV in the neutral pH range. As this is the first study showing Azospirillum brasilense being able to biotransform selenite to selenium particles containing a certain amount of sulfur, even if environmental waters supplemented with selenite were used, they may significantly contribute to the biogeochemical cycling of both elements in soil as well as to their soil-plant transfer. Therefore, microbial biotransformation of selenite under certain circumstances may be used for various bio-remediation and bio-technological applications.

Growth kinetics of grape berry density ("Vitis vinifera" L. 'Black Corinth')

Density is a significant physical property of grape berries depending on their mass, volume and chemical composition - mass concentrations of sugar, water or other chemical compounds. The current research studies the changes of grape berry density for the seedless grape cultivar 'Black Corinth' from anthesis to maturity. It has been found that the change of grape berry density during the growth follows the double sigmoid curve and consists of two growth cycles divided by a lag phase. During the first growth cycle, between anthesis and lag phase, berry density increases from 800 mg·cm-3 to 1020 mg·cm-3. During the lag phase berry growth is delayed and berry density remains almost a constant. At the end of veraison, berry density rose quickly, and during the technological maturity it reached ~ 1120 mg cm-3. The rate of change of berry density has two local maxima coinciding with the middle of the first and second growth cycles of berry development.

A comparative analysis of antibacterial activity, dynamics, and effects of silver ions and silver nanoparticles against four bacterial strains

Although both silver ions and silver nanoparticles (AgNPs) have perfect antibacterial activity, but it was assumed that AgNPs have stronger activity than that of silver ions. In this study, we make a comparative analysis of activity, dynamics, and effects of silver ions and two types of AgNPs against four bacterial strains. The minimum inhibitory concentrations (MICs) of silver ions, AgNPs (I) and AgNPs (II) were 0.5, 1 and 2 μg/mL against E. coli, 1, 2 and 8 μg/mL against P. aeruginosa, 1, 2 and 4 μg/mL against S. aureus, and 1, 2 and 2 μg/mL against S. epidermidis respectively. This experimental results showed that Ag+ have stronger antibacterial activity than that of AgNPs (I) and AgNPs(II). Antibacterial dynamic curves revealed all the silver ions, AgNPs (I), and AgNPs (II) prolonged the growth lag phase of all four bacteria in a concentration-dependent manner. Furthermore, transmission electronic microscopy (TEM) observation showed that a major part of bacterial cells treated with 2 μg/mL of silver ion and AgNPs were destroyed within 5 h. The transmission electron microscopy (TEM) observation indicated that all the silver ions, AgNPs (I), and AgNPs (II) can induce severe damage in bacterial cells. The flagella of bacteria were damaged or even eliminated, which would cause movement disorders. Many holes or gaps were observed on cell surfaces, which would cause the leakage of cytoplasm and macromolecules, and leading to cell death at last. Our results suggested that silver ions have similar action mode and slightly better antibacterial activity than that of AgNPs against bacterial cells.

Combinatorial metabolic engineering of Pseudomonas putida KT2440 for efficient mineralization of 1,2,3-trichloropropane

An industrial waste, 1,2,3-trichloropropane (TCP), is toxic and extremely recalcitrant to biodegradation. To date, no natural TCP degraders able to mineralize TCP aerobically have been isolated. In this work, we engineered a biosafety Pseudomonas putida strain KT2440 for aerobic mineralization of TCP by implantation of a synthetic biodegradation pathway into the chromosome and further improved TCP mineralization using combinatorial engineering strategies. Initially, a synthetic pathway composed of haloalkane dehalogenase, haloalcohol dehalogenase and epoxide hydrolase was functionally assembled for the conversion of TCP into glycerol in P. putida KT2440. Then, the growth lag-phase of using glycerol as a growth precursor was eliminated by deleting the glpR gene, significantly enhancing the flux of carbon through the pathway. Subsequently, we improved the oxygen sequestering capacity of this strain through the heterologous expression of Vitreoscilla hemoglobin, which makes this strain able to mineralize TCP under oxygen-limited conditions. Lastly, we further improved intracellular energy charge (ATP/ADP ratio) and reducing power (NADPH/NADP+ ratio) by deleting flagella-related genes in the genome of P. putida KT2440. The resulting strain (named KTU-TGVF) could efficiently utilize TCP as the sole source of carbon for growth. Degradation studies in a bioreactor highlight the value of this engineered strain for TCP bioremediation.

Process development of human multipotent stromal cell microcarrier culture using an automated high-throughput microbioreactor.

ABSTRACTMicrobioreactors play a critical role in process development as they reduce reagent requirements and can facilitate high‐throughput screening of process parameters and culture conditions. Here, we have demonstrated and explained in detail, for the first time, the amenability of the automated ambr15 cell culture microbioreactor system for the development of scalable adherent human mesenchymal multipotent stromal/stem cell (hMSC) microcarrier culture processes. This was achieved by first improving suspension and mixing of the microcarriers and then improving cell attachment thereby reducing the initial growth lag phase. The latter was achieved by using only 50% of the final working volume of medium for the first 24 h and using an intermittent agitation strategy. These changes resulted in >150% increase in viable cell density after 24 h compared to the original process (no agitation for 24 h and 100% working volume). Using the same methodology as in the ambr15, similar improvements were obtained with larger scale spinner flask studies. Finally, this improved bioprocess methodology based on a serum‐based medium was applied to a serum‐free process in the ambr15, resulting in >250% increase in yield compared to the serum‐based process. At both scales, the agitation used during culture was the minimum required for microcarrier suspension, NJS. The use of the ambr15, with its improved control compared to the spinner flask, reduced the coefficient of variation on viable cell density in the serum containing medium from 7.65% to 4.08%, and the switch to serum free further reduced these to 1.06–0.54%, respectively. The combination of both serum‐free and automated processing improved the reproducibility more than 10‐fold compared to the serum‐based, manual spinner flask process. The findings of this study demonstrate that the ambr15 microbioreactor is an effective tool for bioprocess development of hMSC microcarrier cultures and that a combination of serum‐free medium, control, and automation improves both process yield and consistency. Biotechnol. Bioeng. 2017;114: 2253–2266. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

The CTX-M-14 plasmid pHK01 encodes novel small RNAs and influences host growth and motility.

The dissemination of extended-spectrum β-lactamases (ESBLs) genes among bacteria is commonly achieved by plasmid conjugation. In the last decade, the CTX-M type enzyme was the most widespread and prevalent ESBLs in the world. In Hong Kong and mainland China, among the commonly found CTX-M-carrying plasmids were pHK01 and pHK01-like plasmids, which belong to incompatibility group FII (IncFII). In this work, we studied the physiological effect caused by the pHK01 plasmid in bacterial host Escherichia coli J53. The plasmid did not affect cell growth of the host but reduced their motility. The reduction of host motility was attributed to downregulation of genes that encode the flagellar system. We also identified several plasmid-encoded sRNAs, and showed that the overexpression of one of them, AS-traI, in the presence of pHK01 plasmid shortened the lag phase of host growth. In addition to the study of pHK01 in bacteria, we also developed a fast and incompatibility group-specific curing method using countertranscribed RNA, which could be of general usage for studying plasmid-host interaction in clinical aspects.

Toxicity of nickel to soil microbial community with and without the presence of its mineral collectors-a calorimetric approach.

The toxicity of nickel and three of its main collectors, sodium isopropyl xanthate (SIPX), sodium ethyl xanthate (SEX), and potassium ethyl xanthate (PEX) to soil microbial activity, was analyzed, individually and as a binary combination of nickel and each of the collectors. The investigation was performed through the microcalorimetric analysis method. For the single chemicals, all power-time curves exhibited lag, exponential, stationary, and death phases of microbial growth. Different parameters exhibited a significant adverse effect of the analyzed chemicals on soil microbial activity, with a positive relationship between the inhibitory ratio and the chemical dose (p<0.05 or p<0.01). A peak power reduction level of 24.23% was noted for 50μgg-1 soil in the case of Ni while for the mineral collectors, only 5μgg-1 soil and 50μgg-1 soil induced a peak power reduction level of over 35 and 50%, respectively, in general. The inhibitory ratio ranged in the following order: PEX>SEX>SIPX>Ni. Similar behavior was observed with the mixture toxicity whose inhibitory ratio substantially decreased (maximum decrease of 38.35%) and slightly increased (maximum increase of 15.34%), in comparison with the single toxicity of mineral collectors and nickel, respectively. The inhibitory ratio of the mixture toxicity was positively correlated (p<0.05 or p<0.01) with the total dose of the mixture. In general, the lesser and higher toxic effects are those of mixtures containing SIPX and PEX, respectively.

Short communication: Nutrient consumption patterns of Lactobacillus acidophilus KLDS 1.0738 in controlled pH batch fermentations

This work focused on elucidating the nutrient consumption patterns of Lactobacillus acidophilus to guide the design of media for high-cell-density culture. We investigated the nutrient consumption patterns of L. acidophilus KLDS 1.0738 in chemically defined media in controlled pH batch fermentations. The most abundantly consumed amino acids, vitamins, ions, and purines and pyrimidines were Glu and Gly, pyridoxine and nicotinamide, K+ and PO43-, and guanine and uracil, respectively. The highest consumption rates for amino acids, vitamins, ions, and purines and pyrimidines were Asp and Arg, folic acid and pyridoxine, Fe2+ and Mn2+, and uracil and thymine, respectively. Furthermore, most of the amino acids, as well as guanine, thymine, pyridoxine, folic acid, nicotinamide, Mg2+, PO43-, and K+ had the highest bioavailability from the end of the lag growth phase to the mid-exponential growth phase. The overall consumption of glucose, adenine nucleotides, 2'-deoxyguanosine monohydrate, calcium pantothenate, Fe2+ and Mn2+ decreased with increasing average growth rate, indicating more effective use of these nutritional components at a higher average growth rate, as biomass yield based on nutritional component consumption increased. Our findings help to formulate complex media for high-cell-density cultivation and provide a theoretical basis for L. acidophilus feeding strategies.

Use of enhanced nisin derivatives in combination with food-grade oils or citric acid to control Cronobacter sakazakii and Escherichia coli O157:H7

Cronobacter sakazakii and Escherichia coli O157:H7 are well known food-borne pathogens that can cause severe disease. The identification of new alternatives to heating to control these pathogens in foods, while reducing the impact on organoleptic properties and nutritional value, is highly desirable. In this study, nisin and its bioengineered variants, nisin V and nisin S29A, are used alone, or in combination with plant essential oils (thymol, carvacrol and trans-cinnamaldehyde) or citric acid, with a view to controlling C. sakazakii and E. coli O157:H7 in laboratory-based assays and model food systems. The use of nisin variants (30 μM) with low concentrations of thymol (0.015%), carvacrol (0.03%) and trans-cinnamaldehyde (0.035%) resulted in extended lag phases of growth compared to those for corresponding nisin A-essential oil combinations. Furthermore, nisin variants (60 μM) used in combination with carvacrol (0.03%) significantly reduced viable counts of E. coli O157:H7 (3-log) and C. sakazakii (4-log) compared to nisin A-carvacrol treatment. Importantly, this increased effectiveness translated into food. More specifically, sub-inhibitory concentrations of nisin variants and carvacrol caused complete inactivation of E. coli O157:H7 in apple juice within 3 h at room temperature compared to that of the equivalent nisin A combination. Furthermore, combinations of commercial Nisaplin and the food additive citric acid reduced C. sakazakii numbers markedly in infant formula within the same 3 h period. These results highlight the potential benefits of combining nisin and variants thereof with carvacrol and/or citric acid for the inhibition of Gram negative food-borne pathogens.

Robust parameter identification using parallel global optimization for a batch nonlinear enzyme-catalytic time-delayed process presenting metabolic discontinuities

In this paper, a nonlinear enzyme-catalytic time-delayed switched dynamical system is considered to describe batch culture of glycerol bioconversion to 1,3-propanediol induced by Klebsiella pneumoniae. This system can not only predict the exponential growth phase but also the lag and the stationary growth phases of batch culture since it contains two switching times for representing the starting moment of lag growth phase and the time when the cell specified growth rate reaches the maximum. The biological robustness is expressed in terms of the expectation and variance of the relative deviation. Our aim is to identify the switching times. To this end, a robust parameter identification problem is formulated, where the switching times are decision variables to be chosen such that the biological robustness measure is optimized. This problem, which is governed by the nonlinear system, is subject to a quality constraint and continuous state inequality constraints. Using a hybrid time-scaling transformation to parameterize the switching times into new parameters, an equivalently robust parameter identification problem is investigated. The continuous state inequality constraints are approximated by a conventional inequality constraint, yielding a sequence of approximate robust parameter identification subproblems. The convergence analysis of this approximation is also investigated. Owing to the highly complex nature of these subproblems, a parallel algorithm, based on simulated annealing, is proposed to solve these subproblems. From an extensive simulation study, it is observed that the obtained optimal switching times are satisfactory.