Lag Phase Of Growth Research Articles

Development of a New Modelling Approach and Performance Evaluation of Meta-heuristic Optimization Algorithms for the Prediction of Kinetic Growth Parameters for Pseudomonas spp. in Fish

The main aim of the current work was to build up a new mathematical modelling approach in predictive food microbiology field for the prediction of growth kinetics of microorganisms. For this purpose, the bacterial growth data of Pseudomonas spp. in whole fish (gilt-head seabream) subjected to isothermal and non-isothermal storage temperatures were collected from previously published growth curves. Maximum specific growth rate (1/h) and lag phase duration (h) were described as a function of storage temperature using the direct two-step, direct one-step and inverse dynamic modelling approaches based on various meta-heuristic optimization algorithms. The fitting capability of the modelling approaches and employed optimization algorithms was separately compared, and the one-step modelling approach for the direct methods and the Bayesian optimization method for the used algorithms provided the best goodness of fit results. These two were then further processed in validation step. The inverse dynamic modelling approach based on the Bayesian optimization algorithm yielded satisfactorily statistical indexes (1.02 > Bias factor > 1.09 and 1.07 > Accuracy factor > 1.13), which indicates it can be reliably used as an alternative way of describing the growth behaviour of Pseudomonas spp. in fish in a fast and efficient manner with minimum labour effort.

Candida glycerinogenes Strains Overexpressing Transcription Factors have Improved Furfural Tolerance in Ethanol Production from Non-detoxified Cellulose Hydrolysate.

Cellulose is one of the main raw materials for production of green ethanol, but the presence of the growth inhibitor furfural in non-detoxified lignocellulosic hydrolysates often seriously affects their utilization. In a previous study, we obtained strains of Candida glycerinogenes that were tolerant to furfural, but at concentrations above 2.5g L-1 there was a significant increase in the growth lag phase. In this work, transcription factor genes (SEF1, STB5, CAS5, and ETP1) associated with furfural tolerance were identified and employed to obtain modified strains permitting ethanol fermentation of concentrated and non-detoxified cellulose hydrolysates containing more than 2.5g L-1 furfural. Tolerance to furfural could be increased to 4.5g L-1 by overexpression of either STB5 or ETP1, which have different regulation patterns. Moreover, in non-detoxified and concentrated cellulose hydrolysate, overexpression of ETP1 significantly shortened the growth lag phase and ethanol fermentation time was reduced by 17-20%. In batch fermentations fed with concentrated non-detoxified lignocellulose hydrolysate, ethanol productivity and maximum ethanol concentration reached 2.4g L-1 h-1 and 72.5g L-1, increases of 26.1% and 6.6%, respectively. The results provided a route for the economic use of lignocellulose for chemical production.

Efficient Green Light Acclimation of the Green Algae Picochlorum sp. Triggering Geranylgeranylated Chlorophylls.

In analogy to higher plants, eukaryotic microalgae are thought to be incapable of utilizing green light for growth, due to the “green gap” in the absorbance profiles of their photosynthetic pigments. This study demonstrates, that the marine chlorophyte Picochlorum sp. is able to grow efficiently under green light emitting diode (LED) illumination. Picochlorum sp. growth and pigment profiles under blue, red, green and white LED illumination (light intensity: 50–200 μmol m−2 s−1) in bottom-lightened shake flask cultures were evaluated. Green light-treated cultures showed a prolonged initial growth lag phase of one to 2 days, which was subsequently compensated to obtain comparable biomass yields to red and white light controls (approx. 0.8 gDW L−1). Interestingly, growth and final biomass yields of the green light-treated sample were higher than under blue light with equivalent illumination energies. Further, pigment analysis indicated, that during green light illumination, Picochlorum sp. formed unknown pigments (X1-X4). Pigment concentrations increased with illumination intensity and were most abundant during the exponential growth phase. Mass spectrometry and nuclear magnetic resonance data indicated, that pigments X1-X2 and X3-X4 are derivatives of chlorophyll b and a, which harbor C=C bonds in the phytol side chain similar to geranylgeranylated chlorophylls. Thus, for the first time, the natural accumulation of large pools (approx. 12 mg gDW −1) of chlorophyll intermediates with incomplete hydrogenation of their phytyl chains is demonstrated for algae under monochromatic green light (Peak λ 510 nm, full width at half maximum 91 nm). The ability to utilize green light offers competitive advantages for enhancing biomass production, particularly under conditions of dense cultures, long light pathways and high light intensity. Green light acclimation for an eukaryotic microalgae in conjunction with the formation of new aberrant geranylgeranylated chlorophylls and high efficiency of growth rates are novel for eukaryotic microalgae. Illumination with green light could enhance productivity in industrial processes and trigger the formation of new metabolites–thus, underlying mechanisms require further investigation.

Biofixation of CO2 from a power plant through large-scale cultivation of Spirulina maxima

Using microalgae for biological fixation of CO2 is an attractive option for limiting the amount of CO2 emissions from power plants. In this experiment, Spirulina maxima was successfully grown utilizing direct flue gas emitted from the Ratchaburi Electricity Generating Company Limited, Thailand. The aims of this study were to determine the optimal conditions for large-scale culturing of this alga for maximum biomass as well as CO2 abatement. The results showed that the carbon source of NaHCO3 can be reduced from standard Zarrouk medium of 16.8 down to 8 g/L, producing fresh and dry weight biomass yields of 206 and 21.57 kg, respectively, within 30 days of cultivation. The optimal initial pH was 9.0, thereby reducing the lag phase of algal growth and producing the best fresh weight yield. The natural light intensity (with CO2) prevented stressed and weakened the algal cells, resulting in extending the algal life and producing the highest yields of fresh and dry weight. The morphology of algal cells was examined following culturing at pH 9 and pH10.5. The algal cells produced at pH 9 had a helical shape of the trichome, a separate septum and thylakoids, and the vacuoles were distributed throughout the cell, while algal cells cultured in medium with an initial pH of 10.5 mostly had the straight form, with the septum and thylakoids being unclear. The maximum CO2 level used was 2.5 times base level of the CO2 (141.75 g CO2/h) produced the highest biomass of fresh and dry weights. The optimal culture for CO2 gas was the continuous culture system and using CO2 efficiently, which encouraged algal growth and produced higher biomass yields than those from the batch culture system. There was no significant difference in nutritional value terms for the protein and pigments contents between the algal biomass products in assay with and without CO2 Furthermore, there was no metal contamination in algal biomass. The results indicate that the use of waste CO2 from a power plant can be successfully used for large-scale Spirulina production.

Improving lactic acid production via bio-augmentation with acid-tolerant isolates from source-sorted organic household waste

Biotransformation of source-sorted organic household waste (SSOHW) into lactic acid (LA) could be an attractive waste management approach, converting residual resources to the useful platform chemical and at the same time minimizing pollution. However, nutritional content is not always optimal for the inoculated lactic acid bacteria (LAB) due to the complex composition of SSOHW. In the present study, autochthonous acid-tolerant LA strains were isolated from SSOHW. The isolated strains were used as bio-augmentation inoculum for improving LA production from SSOHW. More specifically, four isolates were identified by sequencing to be Lactobacillus panis DTUAT 01, Lactobacillus pontis DTUAT 02, Lactobacillus oris DTUAT 03, and Lactobacillus reuteri DTUAT 04, which could efficiently convert glucose to LA at pH 4. Kinetic modeling was applied to predict performances and characteristics of the four isolates, and multi-response nonlinear regression analysis was performed on the experimental data to determine the kinetic parameters. The estimated kinetic parameters indicated that strain L. reuteri had the highest maximum growth rate (µmax) and LA yield among the four isolates along with relatively high substrate and product tolerance. Finally, bio-augmentation with strain L. reuteri boosted the LA concentration by 29% compared to abiotic augmentation (i.e., control test). Likewise, autochthonous L. reuteri could quickly adapt to SSOHW as shown by the elimination of its growth lag phase. The findings of this work can contribute to the improvement of fermentative LA production from wastes at acidic conditions.

Inducible boron resistance via active efflux in Lysinibacillus and Enterococcus isolates from boron-contaminated agricultural soil.

Phylogenetically diverse bacteria tolerate high boron concentrations while others require it for metabolic purposes despite the metalloid being toxic beyond a threshold. Boron resistance and plant growth promoting attributes of two bacterial strains, Lysinibacillus sp. OL1 and a novel Enterococcus sp. OL5, isolated from boron-fertilizer-amended cauliflower fields were investigated in this study. OL1 and OL5 grew efficiently in the presence of 210-230mM boron, and resistance was found to be inducible by small amounts of the element: 5 to 50mM boron pre-exposure progressively shortened the lag phase of growth in the presence of 200mM boron. Intracellular boron accumulation was also found to be regulated by the level of pre-exposure: no induction or induction by small amounts led to higher levels of intracellular accumulation, whereas induction by high concentrations led to lower accumulation. These data, in the context of the strains' overall resistance towards 200mM boron, indicated that induction by higher boron concentrations turned potential efflux mechanisms on, while resistance was eventually achieved by continuous cellular entry and exit of the ions. Involvement of solute efflux in boron resistance was corroborated by the genome content of the isolates (genes encoding proteins of the ATP-binding cassette, major facilitator, small multidrug resistance, multi antimicrobial extrusion, and resistance-nodulation-cell division, family/superfamily). Bacteria such as OL1 and OL5, which resist boron via influx-efflux, potentially lower boron bioavailability, and therefore toxicity, for the soil microbiota at large. These bacteria, by virtue of their plant-growth-promoting attributes, can also be used as biofertilizers.

The Effect of Gibberellic Acid on the Production Characteristics and Biochemical Parameters of Tetraselmis Suecica in an Enrichment Culture

The use of gibberellic acid as a stimulator of microalgae growth has beensubstantiatedexperimentally.This research aimed to assess the effect of exposure to a wide range of gibberellic acid concentrations on the growth dynamics ofthe microalgaTetraselmissuecicain an enrichment culture. The duration of the experiments was 14 days. It has been shown that gibberellic acid,atconcentrations of 0.39–3.20× 10−8M, stimulates algaegrowth. In this research, the exposure to gibberellic acid at concentrations of 0.39–3.20 × 10−8M was accompanied by a variation in the pattern of growth curves: the maximum number of cells was recorded on day seven of the experiment. A higher concentration of the phytohormone (3.84 × 10−8М) inhibited the increase inculture density. The growth of theT. suecicaculture in the control group was 332%;the growth of the culture exposed to gibberellic acid at a concentration of 0.39 × 10−8M was1136%. The values of the specific growth rate ofT. suecicawere estimated for different periods of cultivation. On day14 of the experiment, the biochemical composition of microalgae biomass was analyzed.According to the results, gibberellic acid stimulated the accumulation of carbohydrates, proteins, and chlorophyll. Nevertheless, the phytohormone had no effect on lipidaccumulation. An assumption was made thatexposure to low concentrations of phytohormone stimulates the growth of microalgae by reducing the lag phase of growth.
 Keywords: gibberellic acid, microalga, cultivation, lipids, carbohydrates, proteins

Oxidative effects of glyphosate on the lipophobic intracellular environment in the microalgae

The studied hypothesis is that the herbicide glyphosate (GLY) can affect the oxidative balance in the hydrophobic intracellular medium in non-target Chlorella vulgaris cells. Analytical GLY and RoundUp (RUP) supplementation, affected the growth profile. A significant 42% decrease in the cellular biomass in stationary (St) phase was observed in cultures supplemented with either 5 µM of GLY or RUP, as compared to control cultures. The treatment with 0.3 µM of GLY generated non-significant effects on the oxidation rate of 2’, 7’ dichlorofluorescein diacetate (DCFH-DA), neither in exponential (Exp) nor in St phase of development, as compared to control cultures. However, the treatment with either 5 µM GLY or 0.3 and 5 µM RUP lead to a significant decrease in the DCFH-DA oxidation rate, as compared to control cultures. The lipid radical (LR●) generation rate, detected by Paramagnetic Resonance Spectroscopy (EPR), was significantly increased in the presence of RUP, in Lag and Exp phase of growth. The non-enzymatic antioxidants, α-Tocopherol (α-T) and β-Carotene (β-C), are aimed to protect membranes against the damage produced by the radical reactions. The content of β-C was not significantly affected, as compared to control cultures, by any of the treatments, in both growth phases of cellular development. The content of α-T was significantly decreased by the supplementation with either 0.3 or 5 µM of RUP or 5 µM GLY. The LR●/α-T ratio, used as indicator of the oxidative balance in the hydrophobic cellular media, was significantly different between samples obtained from control and RUP-exposed microalgae in both, Exp and St phase of development, with either 0.3 or 5 μM RUP. The data presented here showed evidence that suggested that oxidative balance in the hydrophobic environment was affected by either GLY or RUP.

Comparative genomic identification and characterization of npcRNA homologs in Proteus vulgaris

Proteus vulgaris is a rod-shaped Gram-negative bacterium known to be the member of Enterobacteriaceae that is able to cause disease in human being. Generally, non-protein-coding RNAs (npcRNAs) do not code for proteins, but they play a vital role in gene regulation at the RNA level including pathogenicity. The present study aims at elucidating homologous npcRNAs from other bacteria in Proteus vulgaris. A comparative genomic analysis was carried out to identify npcRNA homolog of other Enterobacteriaceae pathogens in Proteus vulgaris. A total of 231 npcRNAs previously reported in Salmonella typhi, Salmonella typhimurium and Escherichia coli were screened using BLASTn tool against Proteus vulgaris genome. Interestingly, 33 npcRNAs are homologs to Proteus vulgaris. Northern blot analysis of 6 out of 33 npcRNA candidates confirmed their expression and showed that most of them are differentially expressed during lag, exponential and stationary growth phases. This study is the first approach of identification and characterization of npcRNAs in P. vulgaris. Hence, this could be a pioneer study to further validate the regulatory functions of these npcRNAs to fill the gaps in understanding of the pathogenicity of P. vulgaris.

Characterisation of a putative M23-domain containing protein in Mycobacterium tuberculosis.

Mycobacterium tuberculosis, the causative agent of tuberculosis remains a global health concern, further compounded by the high rates of HIV-TB co-infection and emergence of multi- and extensive drug resistant TB, all of which have hampered efforts to eradicate this disease. As a result, novel anti-tubercular interventions are urgently required, with the peptidoglycan component of the M. tuberculosis cell wall emerging as an attractive drug target. Peptidoglycan M23 endopeptidases can function as active cell wall hydrolases or degenerate activators of hydrolases in a variety of bacteria, contributing to important processes such as bacterial growth, division and virulence. Herein, we investigate the function of the Rv0950-encoded putative M23 endopeptidase in M. tuberculosis. In silico analysis revealed that this protein is conserved in mycobacteria, with a zinc-binding catalytic site predictive of hydrolytic activity. Transcript analysis indicated that expression of Rv0950c was elevated during lag and log phases of growth and reduced in stationary phase. Deletion of Rv0950c yielded no defects in growth, colony morphology, antibiotic susceptibility or intracellular survival but caused a reduction in cell length. Staining with a monopeptide-derived fluorescent D-amino acid, which spatially reports on sites of active PG biosynthesis or repair, revealed an overall reduction in uptake of the probe in ΔRv0950c. When stained with a dipeptide probe in the presence of cell wall damaging agents, the ΔRv0950c mutant displayed reduced sidewall labelling. As bacterial peptidoglycan metabolism is important for survival and pathogenesis, the role of Rv0950c and other putative M23 endopeptidases in M. tuberculosis should be explored further.

Co-culturing Hyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 allows sustainable denitrifying activities under marine conditions.

BackgroundHyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities of a methanol-fed, fluidized-bed marine denitrification system. Strain NL23 possesses the complete denitrification pathway, but cannot grow under marine conditions in pure cultures. Strain JAM1 is a marine bacterium that lacks genes encoding a dissimilatory nitrite (NO2−) reductase and therefore cannot reduce NO2−. Here, we report the characterization of some of their physiological traits that could influence their co-habitation. We also perform co-cultures to assess the potential synergy between the two strains under marine and denitrifying conditions.MethodologyAnoxic planktonic pure cultures of both strains were grown with different concentrations of nitrate (NO3−). Anoxic planktonic co-cultures could only be cultured on low NaCl concentrations for strain NL23 to grow. Biofilm co-cultures were achieved in a 500-mL bioreactor, and operated under denitrifying conditions with increasing concentrations of NaCl. NO3− and NO2− concentrations and the protein content were measured to derive the denitrification rates. The concentrations of both strains in co-cultures were determined by quantitative PCR (qPCR). Ectoine concentration was measured by mass spectrometry in the biofilm co-culture. The biofilm was visualized by fluorescence in situ hybridization. Reverse-transcription-qPCR and RNA-seq approaches were used to assess changes in the expression profiles of genes involved in the nitrogen pathways in the biofilm cultures.ResultsPlanktonic pure cultures of strain JAM1 had a readiness to reduce NO3− with no lag phase for growth in contrast to pure cultures of strain NL23, which had a 2-3 days lag phase before NO3− starts to be consumed and growth to occur. Compared to strain NL23, strain JAM1 has a higher µmax for growth and higher specific NO3− reduction rates. Denitrification rates were twice higher in the planktonic co-cultures than those measured in strain NL23 pure cultures. The biofilm co-cultures showed sustained denitrifying activities and surface colonization by both strains under marine conditions. Increase in ectoine concentrations was observed in the biofilm co-culture with the increase of NaCl concentrations. Changes in the relative transcript levels were observed in the biofilm culture with genes encoding NapA and NapGH in strain NL23. The type of medium had a great impact on the expression of genes involved in the N-assimilation pathways in both strains.ConclusionsThese results illustrate the capacity of both strains to act together in performing sustainable denitrifying activities under marine conditions. Although strain JAM1 did not contribute in better specific denitrifying activities in the biofilm co-cultures, its presence helped strain NL23 to acclimate to medium with NaCl concentrations >1.0%.

Microalgal photosynthetic inhibition and mixotrophic growth in Post Hydrothermal Liquefaction Wastewater (PHW)

Hydrothermal Liquefaction converts wet organic biomass into renewable biocrude oil and simultaneously generates a toxic wastewater (PHW) that is, however, rich in nutrients and organic compounds. While inhibition of algae in diluted PHW has been reported, the underlying reasons are still unclear. The present research explores, for the first time, the effects of PHW on the growth, photoautotrophic functions, and biomass characteristics of four freshwater green microalgae. Chlorolobion braunii, Chlorella sorokiniana, Chlorella vulgaris, and Scenedesmus quadricauda were exposed to four PHW concentrations (0, 0.5, 1, and 2% v/v) and their physiological responses comprehensively investigated. Based in pulse amplitude fluorometry (PAM), a general inhibitory effect in photosynthesis was revealed, indicating diminished photoautotrophic activity. Rapid light curve (RLC) showed that different photosynthetic stages were affected and maximum quantum yield decreased, while Gompertz modeling of population growth highlighted differences in both growth rates and lag phases. Physiological adjustment is suggested by reduced cell viability, increased cell size, and modification in pigment profiles. C. sorokiniana stood out based on its limited inhibitory effects and biomass generation in PHW. The combination of diminished photosynthetic performance with increased growth rate affirmed mixotrophy. These findings add to a growing body of literature on the recycling of PHW and similar wastewaters by providing new insights into inhibition mechanisms considering algal photobiology and potential for enhanced biomass production through mixotrophic metabolism.

Bioanalytical insight into the life of microbial populations: A chemical monitoring of ureolytic bacteria growth

In this publication an alternative approach to investigations of bacterial growth is proposed. Contrary to the conventional physical methods it is based on enzyme activity detection. The procedure for real-time and on-line monitoring of microbial ureolytic activity (applied as a model experimental biosystem) in the flow analysis format is presented. The developed fully-mechanized bioanalytical flow system is composed of solenoid micropumps and microvalves actuated by Arduino microcontroller. The photometric detection based on Nessler reaction is performed using dedicated flow-through optoelectronic detector made of paired light emitting diodes. The developed bioanalytical system allows discrete assaying of microbial urease in the wide range of activity up to 5.4 U mL−1 with detection limit below 0.44 U mL−1, a high sensitivity in the linear range of response (up to 200 mV U−1 mL and relatively high throughput (9 detection per hour). The proposed differential procedure of measurements (i.e. a difference between peaks register for sample with and without external addition of urea is treated as an analytical signal) allows elimination of interfering effects from substrate and products of biocatalysed reaction as well as other components of medium used for microbial growth. The developed bioanalytical system was successfully applied for the control of growth of urease-positive bacteria strains (Proteus vulgaris, Klebsiella pneumoniae and Paracoccus yeei) including examination of effects from various microbial cultivation conditions like temperature, composition of culture medium and amount of substrate required for induction of bacterial enzymatic activity. The developed bioanalytical flow system can be applied for metabolic activity-based estimation of parameters of lag and log phases of microbial growth as well as for detection of decline phase.

CsubMADS1, a lag phase transcription factor, controls development of polar eukaryotic microalga Coccomyxa subellipsoidea C-169.

MADS-box transcription factors (TFs) have not been functionally delineated in microalgae. In this study, the role of CsubMADS1 from microalga Coccomyxa subellipsoidea C-169 has been explored. Unlike Type II MADS-box proteins of seed plants with MADS, Intervening, K-box, and C domains, CsubMADS1 only has MADS and Intervening domains. It forms a group with MADS TFs from algae in the phylogenetic tree within the Type II MIKCC clade. CsubMADS1 is expressed strongly in the lag phase of growth. The CsubMADS1 monomer does not have a specific localization in the nucleus, and it forms homodimers to localize exclusively in the nucleus. The monomer has two nuclear localization signals (NLSs): an N-terminal NLS and an internal NLS. The internal NLS is functional, and the homodimer requires two NLSs for specific nuclear localization. Overexpression (OX) of CsubMADS1 slows down the growth of the culture and leads to the creation of giant polyploid multinucleate cells, resembling autospore mother cells. This implies that the release of autospores from autospore mother cells may be delayed. Thus, in wild-type (WT) cells, CsubMADS1 may play a crucial role in slowing down growth during the lag phase. Due to starvation in 2-month-old colonies on solid media, the WT colonies produce mucilage, whereas OX colonies produce significantly less mucilage. Thus, CsubMADS1 also negatively regulates stress-induced mucilage production and probably plays a role in stress tolerance during the lag phase. Taken together, our results reveal that CsubMADS1 is a key TF involved in the development and stress tolerance of this polar microalga.

Environmental Effects of Temperature and Water Potential on Mycelial Growth of Neocosmospora solani and Fusarium spp. Causing Dry Root Rot of Citrus.

This study aimed at evaluating the effect of environmental factors temperature and water potential (Ψw) on the growth of Neocosmospora (Fusarium) solani and three Fusarium species (F. oxysporum, F. equiseti and F. brachygibbosum) associated with citrus dry root rot and to determine the optimum and marginal rate for their growth. The effects of incubation temperature (5-40°C), water potentials (Ψw) (-15.54; -0.67MPa) (0.89-0.995aw) and their interaction (5-30°C) was evaluated on the in vitro radial growth rates of Fusarium spp. and on their lag phase. Secondary models were used to model the combined effect of these factors on radial growth rate. The results underlined a highly significant effects (P < 0.001) of Ψw and temperature and their interactions on radial growth rates and lag phases (λ). The Four studied species were shown tolerant to a temperature of 35°C with an optimum mycelial growth at 30 for N. solani and F. oxysporum and at 25°C for F. equiseti and F. brachygibbosum. However, no growth was observed at both temperatures 5 and 40°C and at Ψw of -9.68MPa (0.93aw). The optimum water potential for growth was ≥-2.69 MPA (>0.98aw). The results from the polynomial model and response surface showing good agreement between observed and predicted values. The external validation on citrus fruit indicated slight differences between predicted and observed values of radial growth. The results of this study will be beneficial for understanding the ecological knowledge of these species and thereby limited preventively their occurrence.

Microbial characterization of the Japanese traditional pickle senmaizuke produced by two different manufacturing processes.

Senmaizuke, a traditional turnip pickle, prepared in Kyoto, Japan, is produced via two types of manufacturing processes: with and without vinegar for fermentation. The aim of this study was to reveal the microbial community and growth behavior in the products and manufacturing processes of two types of senmaizuke. Microbial growth analysis of commercial senmaizuke products showed that both types harbored 102–108 colony forming units (CFU)/g of lactic acid bacteria (LAB) and 102–104 of yeast. The fermented‐type products showed successive growth of LAB during the pickling and ripening processes, whereas LAB in vinegar‐type products showed growth only at the preliminary pickling step before vinegar addition; however, the bacteria were viable at the ripening step. LAB in the vinegar‐type products showed growth when the pH of the pickle was neutralized, indicating that acidification via vinegar retards LAB growth. Metagenomic sequencing showed that the fermented‐type products harbored Lactobacillus, Leuconostoc, and other halophilic and psychrophilic bacteria, with a higher bacterial diversity than in the vinegar‐type products. In the vinegar‐type products, Lactobacillus, Leuconostoc, or both were predominant. Culture tests using LAB isolates and turnip medium suggested that a change in the dominance of Leuconostoc to Lactobacillus members observed in the fermented‐type products during pickling and ripening processes was attributed to the low pH sensitivity of Leuconostoc as well as a relatively long lag phase of growth for adapting to the pickling environment. The findings of this study will be useful for the appropriate quality control and assurance procedure of senmaizuke.

Omnibus Modeling of Listeria monocytogenes Growth Rates at Low Temperatures

Listeria monocytogenes is a pathogen of considerable public health importance with a high case fatality. L. monocytogenes can grow at refrigeration temperatures and is of particular concern for ready-to-eat foods that require refrigeration. There is substantial interest in conducting and modeling shelf-life studies on L. monocytogenes, especially relating to storage temperature. Growth model parameters are generally estimated from constant-temperature growth experiments. Traditionally, first-order and second-order modeling (or primary and secondary) of growth data has been done sequentially. However, omnibus modeling, using a mixed-effects nonlinear regression approach, can model a full dataset covering all experimental conditions in one step. This study compared omnibus modeling to conventional sequential first-order/second-order modeling of growth data for five strains of L. monocytogenes. The omnibus model coupled a Huang primary model for growth with secondary models for growth rate and lag phase duration. First-order modeling indicated there were small significant differences in growth rate depending on the strain at all temperatures. Omnibus modeling indicated smaller differences. Overall, there was broad agreement between the estimates of growth rate obtained by the first-order and omnibus modeling. Through an appropriate choice of fixed and random effects incorporated in the omnibus model, potential errors in a dataset from one environmental condition can be identified and explored.

Mathematical Modeling for the Growth of Salmonella spp. and Staphylococcus aureus in Cake at Fluctuating Temperatures

This study aimed to develop dynamic mathematical models to predict the growth of Salmonella spp. and Staphylococcus aureus in a cake under fluctuating temperatures. Among the nine different types of cakes frequently served during school meals, one type of cake was selected based on bacterial growth and water activity. Cocktails of Salmonella spp. and S. aureus were inoculated in the samples and stored at 4–35 °C for up to 336 h. The growth of Salmonella spp. and S. aureus was observed above 20 and 15 °C, respectively. The bacterial cell counts were fitted in the Baranyi model, and the maximum specific growth rate (μmax; log CFU/g/h) and lag phase duration (LPD; h) were analyzed using a polynomial model as a function of temperature (R2 = 0.968–0.988), and the performance of the developed models was appropriate. Furthermore, dynamic models were developed, and the predictions were acceptable in changing the temperature, indicating that the developed dynamic models can successfully predict the outcomes of Salmonella spp. and S. aureus in cake. These results provide useful information for assessing and managing microbial risk in foods by predicting the behavior of Salmonella spp. and S. aureus in cake, especially in changing temperature.

Predictive Modeling for the Growth of Salmonella spp. in Liquid Egg White and Application of Scenario-Based Risk Estimation.

The objective of the study was to develop a predictive model of Salmonella spp. growth in pasteurized liquid egg white (LEW) and to estimate the salmonellosis risk using the baseline model and scenario analysis. Samples were inoculated with six strains of Salmonella, and bacterial growth was observed during storage at 10–37 °C. The primary models were developed using the Baranyi model for LEW. For the secondary models, the obtained specific growth rate (μmax) and lag phase duration were fitted to a square root model and Davey model, respectively, as functions of temperature (R2 ≥ 0.98). For μmax, the values were satisfied within an acceptable range (Af, Bf: 0.70–1.15). The probability of infection (Pinf) due to the consumption of LEW was zero in the baseline model. However, scenario analysis suggested possible salmonellosis for the consumption of LEW. Because Salmonella spp. proliferated much faster in LEW than in egg white (EW) during storage at 20 and 30 °C (p < 0.01), greater Pinf may be obtained for LEW when these products are stored at the same conditions. The developed predictive model can be applied to the risk management of Salmonella spp. along the food chain, including during product storage and distribution.

Exploring the Role of a Cytokinin-Activating Enzyme LONELY GUY in Unicellular Microalga Chlorella variabilis.

LONELY GUY has been previously characterized in flowering plants to be involved in the direct activation of cytokinins. In this study, the function of the only LONELY GUY gene (CvarLOG1) from unicellular green microalga Chlorella variabilis NC64A has been investigated. CvarLOG1 expressed mainly in the lag and log phases of growth and was confirmed to be a cytokinin-activating enzyme. Overexpression of CvarLOG1 in Chlorella led to extended life in culture by almost 10–20 days, creating a “stay-green” phenotype. In the transformed alga, the cell cycle was lengthened due to delayed entry into the G2/M phase contrary to the known role of cytokinins in stimulating G2/M transition possibly due to excessive levels of this hormone. However, due to the sustained growth and delayed senescence, there was an increase in cell number by 11% and in biomass by 46% at the stationary phase, indicating a potential application for the biofuel industry. The total carbohydrate and lipid yield increased by approximately 30 and 20%, respectively. RNA-Seq-based transcriptomic analysis revealed that the genes associated with light and dark reactions of photosynthesis were upregulated, which may be the reason for the increased biomass. These data show that LOG plays an essential role during the cell cycle and in the functioning of the chloroplast and that the pathway leading to direct activation of cytokinins via LOG is functional in algae.