Anaerobic digestion is a process for resource recovery used to produce biogas. However, research focus has shifted toward producing valuable carboxylates due to low natural gas costs. This study uncoupled methanogenesis from acidogenesis by adding glucose as an external carbon source to maintain low pH and enable stable carboxylate production in bioreactors inoculated with beef cattle waste. The central pattern observed involves glucose utilization converted predominantly to lactate or acetate inhibiting methanogenesis. Shorter retention times (5 vs. 20 days) increased total acid production (53.5 vs. 49.7 g/L), especially lactate and butyrate, while recirculation changed acid profiles toward acetate (about 30-60 % increases). Adjustment of pH from acidic (pH 4.0) to neutral (pH 6.5) enhanced total acid accumulation (up to 58.0 g/L) with acetate as a dominant product, whereas lactate production significantly decreased by about 80 %. Chemical oxygen demand analysis also showed improved carbon conversion with recirculation and pH adjustment, which highlights their role in enhancing carboxylate yields. Microbial community analysis revealed Bacillota (49.4-63.8 %) - Bacilli (33.9-64.7 %) - Bacillus (4.3-35.8 %) as the predominant taxa. Lactic acid bacteria played an important role in early lactate production. Methanogens were detected early, indicating successful inhibition of methanogenesis in later phases. Network analysis further identified Clostridia and Dysgonomonas as major contributors to acetate production and linked Lactiplantibacillus to lactate accumulation. These findings highlight operational parameters in carboxylate profiles and microbial dynamics during anaerobic fermentation with glucose as an external carbon source. We suggest that this mechanism would also apply to any other soluble, rapidly fermented carbohydrate substrate source.

BACKGROUND:Body weight is an important indicator of the overall health and production efficiency in broiler chickens. In broiler houses, body weight of chicks is variable despite the same genetics, hatching and feeding practices within a production system. The objective of this study was to investigate the intestinal microbiota and bile salt hydrolase (BSH) activity in slow and fast growing broiler chickens, which belonged to the 10th and 90th percentile body weight groups, respectively. METHODS:A total of 300 Ross 308 broiler chickens (100 per cohort from three independent cohorts) were selected and mucosal samples from the jejunum, ileum, and cecum were collected at day of arrival, 11 and 25 (n = 450). Then, bacterial counts, 16S rRNA amplicon sequencing, species specific real-time qPCR, as well as BSH activity were analyzed. RESULTS:Results of bacterial counts showed no significant difference between slow and fast growing cohorts (P > 0.05), but they tended to be higher in the slow growing chickens in all measured bacterial groups in cecum. The 16S rRNA amplicon sequencing revealed higher relative abundance of E. coli-Shigella (71.3%-79.8%) at day of arrival, while the most abundant microorganisms at d 25 was Candidatus Arthromitus (slow: 44.5%; fast: 27.4%) in small intestine. qPCR results indicated significant differences in bacterial populations between the slow and fast growing chickens, especially higher total bacteria, Enterococcus, and Clostridium cluster I in the slow growing chickens at d 25. BSH activity was higher in the slow growing chickens than the fast growing chickens [slow: 0.476 ΔOD/protein (μg/mL); fast: 0.258 ΔOD/protein (μg/mL); P < 0.0001], and correlation analysis highlighted associations between BSH activity, body weight, feed intake, body weight gain, and bacterial counts. CONCLUSIONS:We postulate that high total bacteria and Enterococcus abundance are associated with high BSH activity, impacting low feed intake and body weight gain, ultimately resulting in separation into slow and fast growing birds. The findings of this study contribute to understanding the relationship between gut microbiota, BSH activity, and host physiology in broiler chickens, with potential implications for poultry production.

Soil microbiome is responsible for crucial biochemical reactions by interacting with organic compounds in the injection wells in aquifer storage and recovery (ASR) processes. This study investigated seasonal variations in organic compounds and microbial communities in a simulated ASR system. For this purpose, eleven pilot-scale soil columns were operated in ASR mode over 13 months. River water was intermittently fed into the simulated ASR system after sedimentation pretreatment. As a result, the sand ratios in soil texture were slightly increased from 93.2-93.4 % to 96.6-98.1 % while the total organic carbon content in the soil was accumulated and maintained at < 0.2 % during the ASR operation. During the sedimentation process, 9-31 % of dissolved organic carbon (DOC) was removed in the water, while during the storage in the soil column, the DOC in water was fluctuated. However, the DOC concentration (less than 4 mg/L) in the effluent was stably maintained until the end of operational periods. High-throughput sequencing results showed that the predominant phylum was Proteobacteria. Interestingly, at the species level, the microbial community structure clearly shifted with elapsed time and column location; various organic degrading soil microbiome having different physiological characteristics (e.g., oxygen, substrate, and temperature) became predominant. Outcomes of this study suggest that soil microbiome can facilitate the establishment of stable ASR processes for river water treatment by contributing to bioattenuation.

Fermentative volatile fatty acid (VFA) production is a sustainable approach for waste valorization. However, selective product recovery remains challenging due to the range of VFAs produced and their dilute concentrations, requiring energy-intensive purification. Membrane-based electrochemical separations comprise an energy-efficient and continuous platform for small molecule separations. At the same time, there is a lack of suitable ion-exchange membranes for separating between structurally similar organic acids. Here, bicontinuous polyelectrolyte complex (PEC)-layered nanofiltration membranes are designed for the selective recovery of VFAs using redox-mediated electrodialysis. Hydrophobic modification of polyelectrolytes via aza-Michael addition precisely tunes the complexation-induced phase separation behaviors and the assembled nanostructures. Surface-confined layer-by-layer complexation generates a nanoscale bicontinuous PEC active layer with tailored surface properties that is inaccessible through bulk complexation. Redox-mediated electrodialysis using the nanostructured membrane exhibits enhancement of both ion permeability and selectivity toward VFAs, with notable reduction of energy consumption by up to 80% compared to conventional electrodialysis. Treatment of synthetic and cow manure fermentation effluents showcases 2 to 4-fold enrichment of VFAs and simultaneous removal of co-existing organic acids, with an energy consumption as low as 1.5 kWh kg-1. These findings advance the understanding of interfacial complexation-induced phase separation of polyelectrolytes and the development of next-generation nanostructured membranes for multicomponent separations. Bicontinuous polyelectrolyte complex-layered membranes are developed for energy-efficient and selective recovery of volatile fatty acids from fermentation effluents using redox-mediated electrodialysis. Surface-confined complexation of polyelectrolytes concurrent with molecularly tuned phase separation generates a robust bicontinuous layer for enhanced and selective organic anion transport. This work advances the development of next-generation nanostructured membranes for electrochemical multicomponent ion separations. image

Understanding the effects of dosing non-toxigenic Clostridia to cows is rare and has received little attention so far. In the present study, a total of eight lactating dairy cows were divided in two groups: control ( n = 4) or Clostridia challenged (oral supplementation of five diverse strains of Paraclostridium bifermentans , n = 4). Bacterial communities were analyzed by qPCR and next-generation sequencing (NGS) in the buccal mucosa as well as digesta and mucosal samples of the gastrointestinal (GI) tract from rumen to rectum (10 compartments), as well as fecal samples. Transcriptomic analysis of barrier and immune-related gene expression was performed on rumen, jejunum, and liver samples. We observed increased microbial populations with the Clostridial challenge in the buccal tissues and the proximal GI tract (forestomach), correlating with Clostridial loads in the feed. Otherwise, there were no significant differences in microbial populations ( p > 0.05) throughout the distal part of the GI tract. The NGS approach, however, revealed that the Clostridial challenge changed the relative abundance of gut and fecal microbiota. In particular, in the challenge group, no Bifidobacterium was observed in the mucosa-associated microbiota and abundance of Pseudomonadota increased in the feces. These results indicated potential adverse effects of Clostridia to cow health. In general, immune responses to the Clostridial challenge were weak. However, transcriptional analysis revealed the down-regulation of junction adhesion molecule encoding gene (−1.44 of log 2 fold-change), which might impact intestinal permeability.

Introduction:Alicyclobacillus has been isolated from extreme environments such as hot springs, volcanoes, as well as pasteurized acidic beverages, because it can tolerate extreme temperatures and acidity. In our previous study, Alicyclobacillus was isolated during the enrichment of methane oxidizing bacteria from Yellowstone Hot Spring samples.Methods:Physiological characterization and genomic exploration of two new Alicyclobacillus isolates, AL01A and AL05G, are the main focus of this study to identify their potential relationships with a thermoacidophilic methanotroph (Methylacidiphilum) isolated from the same hot spring sediments.Results and discussion:In the present study, both Alicyclobacillus isolates showed optimal growth at pH 3.5 and 55°C, and contain ω-alicyclic fatty acids as a major lipid (ca. 60%) in the bacterial membrane. Genomic analysis of these strains revealed specific genes and pathways that the methanotroph genome does not have in the intermediary carbon metabolism pathway such as serC (phosphoserine aminotransferase), comA (phosphosulfolactate synthase), and DAK (glycerone kinase). Both Alicyclobacillus strains were also found to contain transporter systems for extracellular sulfate (ABC transporter), suggesting that they could play an important role in sulfur metabolism in this extreme environment. Genomic analysis of vitamin metabolism revealed Alicyclobacillus and Methylacidiphilum are able to complement each other's nutritional deficiencies, resulting in a mutually beneficial relationship, especially in vitamin B1(thiamin), B3 (niacin), and B7 (biotin) metabolism. These findings provide insights into the role of Alicyclobacillus isolates in geothermal environments and their unique metabolic adaptations to these environments.

Water scarcity is a severe issue for humans owing to global climate change. Water reuse via reverse osmosis (RO) has been widely used to stabilize water supplies; however, RO membrane fouling increases operational costs and necessitates intermittent operation. The biofouling study on the RO pilot was operated either intermittently or continuously using collected brackish water. The environmental microbiome was incubated under oxic and anoxic conditions to simulate surface and groundwater conditions as the feed water. The least fouling was observed on the RO membrane surface when the RO system was operated intermittently using feed water incubated under anoxic conditions. The microbial results showed that specific biofilm communities were formed on RO membrane surface after brackish water RO (BWRO) operation. The major biofilm-forming bacteria distinctively differed with influent water conditions (i.e., oxic vs. anoxic), whereas they were less different with operational strategies (i.e., continuous vs. intermittent). Intermittent operation could compensate for physical cleaning. However, chemical cleaning showed the most effective results for microbe removal on the membrane surface. Therefore, intermittent operation using anoxic feed water can mitigate fouling formation on RO membranes.

Peach (Prunus persica) is one of the representative climacteric fruits susceptible to environmental stresses, including microbial contamination. This article analyzed major findings from the literature on pre- and post-harvest technologies for maintaining the quality of peach fruit to figure out the strengths and limitations of each treatment strategy. The key implication from studies of pre-harvest agents directly applied to the fruit surface or supplemented as fertilizer was the application of a mixture regarding substances with diverse working mechanisms to prevent excessive use of the agent. The common objectives of previous research on pre-harvest treatments were not only the improvement in the quality of harvested fruit but also the storability during long-term refrigeration due to the short lifespan of peaches. In the case of post-harvest treatments, the efficacy was considerably affected by various determinant factors (e.g., a cultivar of fruit, the sort of technologies, and storage environments), and thus operating conditions optimized for peach fruit were described in this article. Whereas, although the combined treatment of technologies categorized into principles (physical, chemical, and biological approaches) has been adopted to achieve the synergistic effect, undesirable antagonistic effects (i.e., the inhibition of efficacies expectable from singular treatments) were also reported to highlight the importance for exploring adequate treatment conditions.

Bacterial co-culture studies using synthetic gut microbiomes have reported novel research designs to understand the underlying role of bacterial interaction in the metabolism of dietary resources and community assembly of complex microflora. Since lab-on-a-chip mimicking the gut (hereafter “gut-on-a-chip”) is one of the most advanced platforms for the simulative research regarding the correlation between host health and microbiota, the co-culture of the synthetic bacterial community in gut-on-a-chip is expected to reveal the diet–microbiota relationship. This critical review analyzed recent research on bacterial co-culture with perspectives on the ecological niche of commensals, probiotics, and pathogens to categorize the experimental approaches for diet-mediated management of gut health as the compositional and/or metabolic modulation of the microbiota and the control of pathogens. Meanwhile, the aim of previous research on bacterial culture in gut-on-a-chip has been mainly limited to the maintenance of the viability of host cells. Thus, the integration of study designs established for the co-culture of synthetic gut consortia with various nutritional resources into gut-on-a-chip is expected to reveal bacterial interspecies interactions related to specific dietary patterns. This critical review suggests novel research topics for co-culturing bacterial communities in gut-on-a-chip to realize an ideal experimental platform mimicking a complex intestinal environment.

The solar-powered membrane distillation (SPMD) process can improve the energy efficiency by using solar energy as a heat source. However, the SPMD process can only be intermittently operated due to the variation of the daily solar irradiation. In this study, effects of intermittent modes (IMs with/without temperature variations (IM-1/IM-2)) and continuous mode (CM) on scaling and wetting are investigated according to three types of shutdown protocols (P1: non-draining, P2: draining, P3: flushing after draining). A direct contact membrane distillation coupled with a real-time visualization system using the normalized light intensity and SEM-EDS are used for analysis of the MD performance in each condition. Consequently, scaling and wetting tendencies of SPMD at P3 are lowest among the tested shutdown protocols. Furthermore, scaling and wetting in CM and IM (IM-1 and IM-2) at P3 show low differences, indicating that shutdown protocols have a more profound effect than temperature variations and operation mode.

Verrucomicrobiotal methanotrophs are thermoacidophilic methane oxidizers that have been isolated from volcanic and geothermal regions of the world. We used a metagenomic approach that entailed obtaining the whole genome sequence of a verrucomicrobiotal methanotroph from a microbial consortium enriched from samples obtained from Nymph Lake (89.9 °C, pH 2.73) in Yellowstone National Park in the USA. To identify and reconstruct the verrucomicrobiotal genome from Illumina NovaSeq 6000 sequencing data, we constructed a bioinformatic pipeline with various combinations of de novo assembly, alignment, and binning algorithms. Based on the marker gene (pmoA), we identified and assembled the Candidatus Methylacidiphilum sp. YNP IV genome (2.47 Mbp, 2392 ORF, and 41.26% GC content). In a comparison of average nucleotide identity between Ca. Methylacidiphilum sp. YNP IV and Ca. Methylacidiphilum fumariolicum SolV, its closest 16S rRNA gene sequence relative, is lower than 95%, suggesting that Ca. Methylacidiphilum sp. YNP IV can be regarded as a different species. The Ca. Methylacidiphilum sp. YNP IV genome assembly showed most of the key genes for methane metabolism, the CBB pathway for CO2 fixation, nitrogen fixation and assimilation, hydrogenases, and rare earth elements transporter, as well as defense mechanisms. The assembly and reconstruction of a thermoacidophilic methanotroph belonging to the Verrucomicrobiota phylum from a geothermal environment adds further evidence and knowledge concerning the diversity of biological methane oxidation and on the adaptation of this geochemically relevant reaction in extreme environments.

Photo-Fenton-active (PFA) membranes can provide an effective strategy to control various organic pollutants in water purification applications by simultaneously promoting photocatalysis and Fenton reaction. However, the fabrication of PFA membranes with high reactivity and stability remains a significant technical challenge. In this study, we present a simple and versatile technique to fabricate a PFA membrane via in-situ functionalization of polypyrrole (PPy, i.e., photothermal catalyst) and zerovalent iron (ZVI, i.e., photo-Fenton catalyst) nanoparticles. The proposed one-pot two-step protocol involves vapor phase polymerization of pyrrole and subsequent reduction of iron source, which successfully creates a thin catalyst coating on a porous substrate without reducing membrane porosity. The PFA membrane with the dual PPy/ZVI catalyst layer showed high surface wettability, photothermal performance, and photo-Fenton reactivity, thereby enabling effective control of organic contaminants. Specifically, the PFA membrane exhibited near complete removal (>97%) of organic dye molecules in five repeated photo-Fenton-oxidation cycles. We found that robust anchoring of the ZVI within the PPy layer is an important factor for stable photo-Fenton activity of the PFA membrane. The enhanced anti-fouling and self-cleaning properties of the PFA membrane were further demonstrated in oil-emulsion filtration experiments with multiple cycles. Our innovative fabrication approach can serve as a versatile platform for the development of highly reactive and stable PFA membranes for a wide range of solar-driven water treatment applications.

Salted napa cabbage is the most important ingredient of kimchi. Currently, people have started to prepare ready-to-use salted napa cabbage at home. This study focused on this trend by investigating consumers' beliefs, opinions, and actual use of the products by conducting a telephone survey (895 female consumers) and face-to-face interviews (n 1/4 514) in 2016 and a telephone survey (n 1/4 200) in 2021. Most respondents (93 and 91% in 2016 and 2021, respectively) answered that convenience was the main reason for using salted napa cabbage. Regarding consumption behavior, 22 and 16% of the respondents in each year used salted napa cabbage after storing it for more than 24 h. In particular, 85 and 91% of consumers stored the product at room temperature, and 60 and 58% used it without washing, which could affect the quality of the food, as microorganisms could multiply during the storage. Inappropriate handling increased by age group, especially in 2021 (P, 0.05). In the query on satisfaction after using the products, 85 and 80% of respondents were satisfied because the product was convenient (54%) and hygienic (17%). Conversely, the respondents who were not satisfied with the products did not like the degree of salting and unhygienic status of the products. The majority (93 and 80%) of consumers preferred buying salted napa cabbage again because of its convenience. Although consumers thought that hygiene and quality were important factors, many respondents (83 and 72%) were not aware of foodborne illnesses associated with kimchi. Nevertheless, consumers intended to pay more for safe, salted napa cabbage (72 and 76%). The results of this study provide useful and credible data for understanding the factors affecting consumers' consumption and general beliefs and opinions on the use of salted napa cabbage, especially for food safety management.

Purpose: Plant-derived essential oils are widely used as pharmacological drugs and fragrances. In this study, 16 natural essential oils and 14 single fragrance ingredients were investigated for their potential as antimicrobial agents.Methods: Paper disk diffusion assays were performed to assess the antimicrobial activities of the essential oils and single fragrance ingredients against Staphylococcus aureus , Staphylococcus epidermidis , Escherichia coli , Pseudomonas aeruginosa , Bacillus subtilis , Candida albicans , Aspergillus niger , and Malassezia furfur.Results: Cinnamon bark and neroli essential oils, which contain linalool, exhibited significant antimicrobial activities against S. aureus and C. albicans. The single fragrance ingredients citral, D -limonene, and hydroxycitronellal exhibited antibacterial effects against S. aureus and C. albicans. Cinnamon bark oil also showed good antifungal effects against A. niger and M. furfur . Clove essential oil, which contains eugenol as the main ingredient, showed antifungal effects against M. furfur, and aurantiol also exhibited antibacterial effects against M. furfur . S. aureus , E. coli, and P. aeruginosa , which are regulated in distributed cosmetics, were susceptible to the effects of cinnamon bark oil and phenylethyl alcohol. Bergamot and lavender oils exerted good antibacterial effects against S. aureus , but not S. epidermidis . In addition, rosemary oil, rose absolute, and phenylethyl alcohol, which showed good antifungal effects against C. albicans , had no or little antibacterial effect on S. epidermidis .Conclusion: Our results suggest that essential oils can be used as alternative preservatives and antimicrobial agents for atopic dermatitis and dandruff.

Escherichia coli O157:H7 EDL933 exposed to low-shear modeled microgravity (LSMMG) and normal gravity (NG) was used for a transcriptomic analysis. The modified Gompertz model (R-2 = 0.81-0.99) showed an increased growth rate of E. coli O157:H7 under LSMMG. The mechanism of this active growth was associated with highly upregulated genes in nutrient and energy metabolism, including the TCA cycle, glycolysis, and pyruvate metabolism. Green fluorescent protein-labeled E. coli O157:H7 also formed significantly thick biofilms (fluorescent unit: NG, 1,263; LSMMG, 1,533; P = 0.0473) under LSMMG, whereas bacterial mobility decreased slightly (P = 0.0310). The transcriptomic analysis revealed that genes encoding glycogen biosynthesis (glgCAP operon) were upregulated (1.40 to 1.82 of log fold change [FC]) due to the downregulation of csrA (2.17 of log FC), which is the global regulator of biofilm formation of E. coli. We also identified 52 genes in E. coli O157:H7 EDL933 that were involved in the secretion pathway, 32 of which showed >= 2-fold significant changes in transcription levels after cultivation under LSMMG. Notably, all downregulated genes belonged to the type III and VI secretion systems, indicating that host cell contact secretion was dysregulated in the LSMMG cultures compared to the NG cultures. LSMMG also stimulates the pathogenicity of E. coli O157:H7 via transcriptional upregulation of Shiga toxin 1 (1.36 to 2.81 log FC) and toxin HokB (6.1 log FC). Our results suggest LSMMG affects bacterial growth, biofilm formation, and E. coli O157:H7 pathogenicity at some transcriptional levels, which indicates the importance of understanding biological consequences.

Aeromonas hydrophila is an emerging foodborne pathogen capable of causing human gastroenteritis, and the main reservoir is the aquatic environment. In this study, the prevalence and virulence of A. hydrophila in seafoods and ready-to-eat (RTE) sushi distributed in various conditions (refrigerated, dried, or frozen) or seasons was investigated. Strains were isolated from seafood (refrigerated or frozen oysters, sashimi, and processed fish; n = 333) and RTE sushi (n = 88) samples collected in South Korea and then genetically analyzed for gastroenteritis-related virulence genes (aer, ast, and alt). Raw oysters showed the highest prevalence of A. hydrophila (57.1%; 47/91) among all seafoods. Among the sashimi samples, flatfish sashimi (54.8%; 34/62) and salmon sushi (51.4%; 18/ 35) were the most prevalent. A. hydrophila was not detected in the oysters or anchovies distributed as either frozen or dried products. Seasonal investigations of sashimi and sushi showed that the summer prevalence of A. hydrophila with putative virulence genes was significantly lower in sashimi but highest in sushi. These results indicated that sushi could have been contaminated from several sources during the manufacturing or distribution processes. Significant correlations among the prevalence of putative virulence genes were confirmed, although no combination of genes presented a Phi correlation coefficient above 0.5 (0.26-0.43). To our knowledge, this is the first study to investigate the prevalence of A. hydrophila in various types of retail seafoods and RTE sushi in the East Asia region and then relate the prevalence to the distribution conditions of the samples. This study provides background information on the level of potential risk posed by A. hydrophila in retail seafoods and RTE sushi.

Acid resistance is critical for the survival of Escherichia coli O157:H7 in acidic environments. The representative space environment microgravity is known to have a great impact on bacteria, but the acid stress response of E. coli O157:H7 under microgravity conditions remains unclear. Here, we show that the acid resistance of sbacteria is altered by the upregulation of related resistance systems. All tested E. coli O157:H7 strains (ATCC 35150, 43889, 43890, and 43895) survived better in acidified Luria-Bertani medium (pH 3.5) under low-shear modeled microgravity (LSMMG) than under normal gravity (NG, counterpart condition). For example, after 72 h of cultivation under acidic conditions, bacterial populations in the LSMMG cultures reached 5.2-6.7 log CFU/ml, while those in the NG cultures reached 2.4-5.6 log CFU/ml. Our transcriptomic analysis studies on E. coli O157: H7 under LSMMG conditions also provided supportive data of the increase in the acid stress response, with a 2.18 to 3.44 log(2) fold change in the acid resistance system 1 (rpoS) and 2 (gad) and chaperone related genes (hdeA and hdeB). Comparing D-values before and after acid shock at pH 3.5, the increase in thermal cross-protection power was more remarkable in the LSMMG cultures than in the NG cultures. In the case of E. coli O157:H7 ATCC 35150, the D-values in the LSMMG and NG cultures at 55 degrees C after acid shock increased by 17.1 and 10.8 min, respectively, compared to the control. Our findings illustrated that simulated microgravity impacts the acid resistance of E. coli O157:H7 as well as the acquisition of thermal cross-protection power, suggesting that alterations in bacterial responses to the space environment could be a health threat.

Customized cosmetics immediately made on the spot are now in the market. The present study surveyed 1084 consumers to obtain general insights into their perceptions and perspectives on this novel type of cosmetic. Over half of the total respondents (57.2%) answered they were likely to purchase customized cosmetics; however, a large proportion of consumers also thought microbiological (59.2%)/chemical safety (69.4%) of cosmetics were not good. This reflects consumer anxiety regarding safety issues concerning the customized cosmetics. Even customized cosmetics are regulated by the cosmetic act in each country (i.e., the Cosmetic Act in the Republic of Korea, the Federal Food, Drug, and Cosmetic Act in the USA, and EC Cosmetic Regulation 1123/2009 in Europe), there have been no specific regulations for customized cosmetics made on the spot worldwide so far. To dispel consumer concerns and establish a principled market for the new cosmetics in the field, proper management plans should be established based on consumer surveys. This study indicated that consumers thought it was important to manage the facility/equipment and safety of raw materials (19.7%, each). We believe this study provides a valuable resource for understanding consumers' perceptions and requirements on customized cosmetics, which contributes to establishing future regulations and guidelines.

Microfluidic labchips have achieved much advancement in the molecular diagnosis of foodborne pathogens. Whereas difficulties in the flow control during the transportation of liquid fluids can occur and should be overcome. Manipulations of reaction temperature and the complex procedures from sample pre-treatment to analysis in a single chip device are major obstacles for the on-site application. Thus, the efficient temperature control of samples without any flow of reaction fluids in microfluidic channels of plastic chip and the simplest protocol omitting post-enrichment processing steps may overcome these limitations represented by the stability and the complexity, respectively. This study aims to develop a novel type of labchip and thermocycler specialized for the gene amplification in microfluidic channels and to evaluate the detectability by sensing the minimum recoverable level of Cronobacter in powdered infant formula (PIF). We developed a thermocycling device accelerating reactions through dual heating-blocks optimized to control temperatures of samples in microfluidic-channels by direct contact with labchip sequentially and repetitively. The structural design of microfluidic channels was to eliminate interference factors associated with the optical detection of fluorescent signals (without distortion due to air bubbles in the reaction chamber). To improve the applicability, a portable device and simplified operation to allow direct loading of samples in the chip without post-enrichment procedures were also adopted. Detection performance was evaluated by a sensitivity/specificity tests using 50 isolates of Cronobacter. Cross-reactivity tests for non-Cronobacter organisms and gDNA [human, raw materials of PIF (cow, soybean)] showed that there was no interference-factor causing false-positive results. In terms of the applied research conducted by using PIF, the enrichment of samples without broth medium (distilled water) displayed outstanding performance and 12 h of incubation facilitated detecting target at concentration as low as 1 CFU/300 g PIF (as initial contamination level) without post-enrichment treatment. Validation of the operation conditions using 30 commercial PIF products was also consistent. The present study presents a novel approach of microfluidic technology with perspective to not only the performance and the practicability [easy-to-implement protocol, portable materials, cost-effectiveness (the use of a miniaturized plastic chip requires a minimum level of materials)] for on-site diagnosis.