Microbiological Research Research Articles

Overview of Foodborne viruses: Important viruses, outbreaks, health concerns, food Handling and fresh produce

Foodborne viruses can transmit through food in lots of ways including consuming items of animal origin containing zoonotic viruses, consuming contaminated food handled by infected food workers, and consuming contaminated food produced by humans. Viral foodborne illnesses are now a major contributor to all foodborne illness reports in recent years and are seen as a rising issue to the public health of humans and animals. Noroviruses and hepatitis A viruses were shown to be predominantly linked to the food-handler transmission and sewage-contaminated foods, according to microbiological research. In order to facilitate source attribution and identify risk preventive measures, routine, standard surveillance of viral outbreaks, and surveillance of virus occurrence in food products, combined with systematic strain typing, food and clinical microbiologists, would be advocated.

Coral holobiont research needs spatial analyses at the microbial scale.

Coral holobiont research needs spatial analyses at the microbial scale.

Serotype-identifying ions in Listeria monocytogenes using matrix-associated laser desorption ionization-time of flight mass spectrometry

Listeria monocytogenes is a foodborne pathogen that can cause a potentially life-threatening infection, and almost all cases of human listeriosis are caused by L. monocytogenes isolates in serotypes 1/2a, 1/2b, 1/2c, and 4b. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a rapid, accurate, and high-throughput tool for clinical diagnosis and microbiological research. In the current study, we examined the potential of MALDI–TOF MS for rapid identification of the foodborne pathogen L. monocytogenes and to identify high-risk serotypes. To achieve this, MALDI–TOF MS was applied to 50 L monocytogenes strains. All strains were identified as L. monocytogenes species based on pattern matching against reference spectra for the species. Importantly, 83 specific mass ions were consistently and uniquely found in high-risk L. monocytogenes serotypes 1/2a, 1/2b, 1/2c, and 4b. These 83 mass ions were also unique to specific combinations of these serotypes, which enabled specific identification of these four serotypes using MALDI Biotyper analysis. Hence, this method shows potential for using MALDI-TOF MS for the rapid identification of L. monocytogenes species and to discriminate high-risk L. monocytogenes serotypes through specific serotype-specific biomarker ions.

Investigation of Microbial Cooperation via Imaging Mass Spectrometry Analysis of Bacterial Colonies Grown on Agar and in Tissue During Infection.

Understanding the metabolic consequences of microbial interactions that occur during infection presents a unique challenge to the field of biomedical imaging. Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry represents a label-free, in situ imaging modality capable of generating spatial maps for a wide variety of metabolites. While thinly sectioned tissue samples are now routinely analyzed via this technology, imaging mass spectrometry analyses of non-traditional substrates, such as bacterial colonies commonly grown on agar in microbiology research, remain challenging due to the high water content and uneven topography of these samples. This paper demonstrates a sample preparation workflow to allow for imaging mass spectrometry analyses of these sample types. This process is exemplified using bacterial co-culture macrocolonies of two gastrointestinal pathogens: Clostridioides difficile and Enterococcus faecalis. Studying microbial interactions in this well-defined agar environment is also shown to complement tissue studies aimed at understanding microbial metabolic cooperation between these two pathogenic organisms in mouse models of infection. Imaging mass spectrometry analyses of the amino acid metabolites arginine and ornithine are presented as representative data. This method is broadly applicable to other analytes, microbial pathogens or diseases, and tissue types where a spatial measure of cellular or tissue biochemistry is desired.

Plasmids for Controlled and Tunable High-Level Expression in E. coli.

Controlled gene expression is crucial for engineering bacteria for basic and applied research. Inducible systems enable tight regulation of expression, wherein a small-molecule inducer causes the transcription factor to activate or repress transcriptional initiation. The T7 expression system is one of the most widely used inducible systems, particularly for high overexpression of proteins. However, it is well known that the highly active T7 RNA polymerase (RNAP) has several drawbacks, including toxicity to the host and substantial leaky expression in the absence of an inducer. Much work has been done to address these issues; current solutions require special strains or additional plasmids, making the system more complicated and less accessible. Here, we challenge the assumption that the T7 expression system is the best choice for obtaining high protein titers. We hypothesized that expression from strong inducible promoters expressed from high-copy plasmids could compete with expression levels obtained from T7 RNAP but that such promoters would possess improved control of transcription. Employing inducible systems from a toolbox we developed previously, we demonstrate that our plasmids consistently give higher outputs and greater fold changes over basal expression than the T7 system across rich and minimal media. In addition, we show that they outperformed the T7 system when we used an engineered metabolic pathway to produce lycopene. IMPORTANCE Genetic systems for protein overexpression are required tools in microbiological and biochemical research. Ideally, these systems include standardized genetic parts with predictable behavior, enabling the construction of stable expression systems in the host organism. Modularity of a genetic system is advantageous, so that the expression system can be easily moved into a host that best suits the needs of a given experiment. The T7 expression system lacks both predictability and stability and requires special host strains to function. Despite these limitations, it remains one of the most popular systems for protein overproduction. This study directly compared the T7 system to four inducible systems from our broad-host-range plasmid toolbox and demonstrated these alternative expression systems have distinct advantages over the T7. The systems are entirely plasmid-based and not constrained to a specific bacterial host, expanding the options for high-level protein expression across strains.

Biopharmaceutical and microbiological research of fennel fruit infusions

Aim. To develop and study physico-chemical, pharmaco-technological, pharmacognostic and microbiological quality indicators of infusions obtained by extraction of the plant raw material of fennel fruits of different degrees of grinding and from different types of packaging. Materials and methods. The study objects were model samples of fennel fruits of different degrees of grinding and water extracts obtained on their basis and from different types of packaging. Water extracts were prepared according to the SPhU method and the patient information leaflet on fennel fruits. The study of pH of solutions, extractives, identification of the main biologically active substances (BAS), and microbiological purity of water extracts were performed by the SPhU methods; the quantitative determination of the essential oil was conducted by direct alkalimetric titration with 0.01 mol/l NaOH solution in the presence of methylene blue and phenolphthalein indicators according to the Guidelines of the Ministry of Health of Ukraine. Results and discussion. Extractives in model samples of infusions obtained from filter bags vary from 0.43 to 0.48 % without filter bags (samples 1, 3, 5, 7) – from 0.52 to 0.55 %, and its amount is the lowest in sample 9 – 0.37 %. However, the essential oil content is the highest in sample 9, these are whole fennel fruits from the pack (0.15 %); slightly higher indicators are in model samples without filter bags (samples 2, 4, 6, 8). The pH value is slightly higher in infusions obtained from filter bags than without them (in the range of 6.61-6.70 and 6.22-6.25, respectively). However, the infusion obtained from a nylon pyramid bag (sample 7) had a pH value close to infusions obtained from the fennel medicinal raw material without packaging (6.25). The presence of infusions of anethole and terpenoids in model samples has been proven by thin layer chromatography. The microbiological studies conducted have shown that the use of the method of making water extracts from the patient information leaflet allows obtaining conditionally sterile infusions. Conclusions. The indicators studied (transparency, color, odor, taste, extractives, identification and quantitative determination of BAS) can be used in the analysis of water extracts from the medicinal plant raw material produced in filter bags.

A Machine Learning-Empowered Workflow to Discriminate Bacillus subtilis Motility Phenotypes

Bacteria that are capable of organizing themselves as biofilms are an important public health issue. Knowledge discovery focusing on the ability to swarm and conquer the surroundings to form persistent colonies is therefore very important for microbiological research communities that focus on a clinical perspective. Here, we demonstrate how a machine learning workflow can be used to create useful models that are capable of discriminating distinct associated growth behaviors along distinct phenotypes. Based on basic gray-scale images, we provide a processing pipeline for binary image generation, making the workflow accessible for imaging data from a wide range of devices and conditions. The workflow includes a locally estimated regression model that easily applies to growth-related data and a shape analysis using identified principal components. Finally, we apply a density-based clustering application with noise (DBSCAN) to extract and analyze characteristic, general features explained by colony shapes and areas to discriminate distinct Bacillus subtilis phenotypes. Our results suggest that the differences regarding their ability to swarm and subsequently conquer the medium that surrounds them result in characteristic features. The differences along the time scales of the distinct latency for the colony formation give insights into the ability to invade the surroundings and therefore could serve as a useful monitoring tool.

New Advances, Challenges and Opportunities in Forensic Applications of Microbiomics.

The succession of microbiota is closely associated with several essential factors, including race, sex, health condition, lifestyle, postmortem interval, etc., and it has great potential application value in forensic medicine. This paper summarizes recent studies on the forensic applications of the microbiome, including individual identification, geographical feature identification, origin identification of the tissue or body fluid, and postmortem interval estimation, and introduces the current machine learning algorithms for microbiology research based on next-generation sequencing data. In addition, the current problems facing forensic microbiomics such as the extraction and preservation of samples, construction of standardization and database, ethical review and practical applicability are discussed. Future multi-omics studies are expected to explore micro ecosystems from a comprehensive and dynamic perspective, to promote the development of forensic microbiomics application.

Recent Developments in Microbiology

Research in microbiology is a dynamic and exciting field, and this review article outlines some recent significant developments within microbiology including using shotgun metagenomics to uncover the connection between diet and lifestyle and superbug infections, the current state of Ebola virus diagnostic tests, and the potential of treating superbug infections via innovative phage virus therapy as an alternative to antibiotics. This review also explores two studies using the CRISPR gene editing technique, which has been a revolution within microbiology. The first study undergoes genetic engineering of potatoes to improve their nutritional and industrial applications, which has important implications in improving food stability as well as bolstering production. The second study utilizes CRISPR to engineer bacteria to kill themselves on command, which is important to prevent the excessive spread bacteria that are used for a variety of applications from eating plastic waste in the environment to treating the diseased human gut. Engineering these so-called ‘kill switch’ allows these bacteria to be utilized for their intended purpose without growing uncontrollably in the environment or in the gut.

Comparison Between Fuzzy Logic and PID Controllers in Temperature Control of Laboratory Incubator

Incubators are very essential equipment that finds wide application in the fields of medical Laboratory and microbiological research organization. This is because Incubator monitors and maintains conducive environment that is suitable for culturing, or growing of micro bacterial and enzymes, for the purpose of clinical diagnoses in Health centres, or process development in manufacturing industries. In this paper, a transfer function model for an incubator was developed upon which a designed Fuzzy logic controller acts to control the incubator Temperature, by computing appropriate Voltage. A PID (Proportional Integral Derivative) controller was also designed and tested on the same model. Findings were that the fuzzy logic controller tracked the optimum Temperatures for culturing of Mesophilic and thermophilic bacterial at 37 0C and 54.94 0C respectively. In comparison with the Fuzzy logic controller, the PID controller tracked at 37 0C and 55 0C for the two selected set points. In addition, the fuzzy logic controller gave faster rise time ( 3.30 mS, 3.45 mS and 13.72 mS) and settling time ( 1.60 mS, 2.04 mS and 5.55 mS ) as compared to PID controller which presented longer rise time (53.61 mS, 53.68 mS and 53.62) and settling time ( 254 mS, 336 mS and 345 mS) for the given set points respectively.

The CINAMR (Clinical Information Network-Antimicrobial Resistance) Project: A pilot microbial surveillance using hospitals linked to regional laboratories in Kenya: Study Protocol.

Background: Antimicrobial resistance (AMR) is a global threat and is thought to be acute in low-and middle-income country (LMIC) settings, including in Kenya, but there is limited unbiased surveillance that can provide reliable estimates of its burden. Current efforts to build capacity for microbiology testing in Kenya are unlikely to result in systematic routine microbiological testing in the near term. Therefore, there is little prospect for microbiological support to inform clinical diagnoses nor for indicating the burden of AMR and for guiding empirical choice of antibiotics. Objective: We aim to build on an existing collaboration, the Clinical Information Network (CIN), to pilot microbiological surveillance using a 'hub-and-spoke' model where selected hospitals are linked to high quality microbiology research laboratories. Methods: Children admitted to paediatric wards of 12 participating hospitals will have a sample taken for blood culture at admission before antibiotics are started. Indication for blood culture will be a clinician's prescription of antibiotics. Samples will then be transported daily to the research laboratories for culture and antibiotic susceptibility testing and results relayed back to clinicians for patient management. The surveillance will take place for 6 months in each hospital. Separately, we shall conduct semi-structured interviews with frontline health workers to explore the feasibility and utility of this approach. We will also seek to understand how the availability of microbiology results might inform antibiotic stewardship, and as an interim step to the development of better national or regional laboratories linked to routine surveillance. Conclusions: If feasible, this approach is less costly and periodic 'hub-and-spoke' surveillance can be used to track AMR trends and to broadly guide empirical antibiotic guidance meaning it is likely to be more sustainable than establishing functional microbiological facilities in each hospital in a LMIC setting.

Assessment of the quality and nutritional value of organic dried sausages

Organic production, which is focused on the production of high-quality, high-nutritional and environmentally friendly food products, is becoming increasingly relevant in developed countries, in particular, Ukraine. The purpose of the study is to determine the quality and nutritional value of organic dried sausages. For its implementation, physico-chemical, microbiological, organoleptic, and statistical research methods and approaches to abstraction, synthesis, analysis, systematisation, and generalisation of data are used. Three types of sausages are selected for the study: Sample 1, Sample 2, and Sample 3. As a result of the study, it is identified that all indicators of the investigated samples are controlled and meet the requirements of regulatory documents. During the organoleptic assessment, it is identified that all the investigated samples had a dry surface without damage, an elastic consistency, and a pleasant smell with the aroma of spices. The difference was observed only in colour, it was from dark pink to dark red, which is due to different recipes of sausages. Microbiological analysis of the products shows that there was no pathogenic microflora in all samples. According to the results of the physico-chemical analysis, it is identified that the content of moisture, salt, and sodium nitrite was within the normal range. The difference was observed in protein and fat content. The first sample of sausage showed the lowest protein content (14%), while the second and third samples showed 16.2% and 19.2%, respectively. Substantial differences were also observed in the fat content of the finished product. The lowest level of this indicator was in Sample 3 and was 6.4%, which is 22.8% and 13.6% lower compared to Sample 1 and Sample 2. This difference in protein content is due to the different composition of raw materials of the investigated samples. The lowest caloric content (134 kcal) is identified in Sample 3 sausage, which had the highest protein levels and the lowest fat content. The results obtained practically allow assessing the dietary characteristics of the investigated assortment and orient the consumer in choosing a product in accordance with their individual preferences and needs, characteristics, and nature of the diet

The Future of Clean Computing May Be Dirty

The emergence of the Internet of Things and pervasive sensor networks have generated a surge of research in energy scavenging techniques. We know well that harvesting RF, solar, or kinetic energy enables the creation of battery-free devices that can be used where frequent battery changes or dedicated power lines are impractical. One unusual yet ubiquitous source of power is soil (earth itself) - or more accurately, bacterial communities in soil. Microbial fuel cells (MFCs) are electrochemical cells that harness the activities of microbes that naturally occur in soil, wetlands, and wastewater. MFCs have been a topic of research in environmental engineering and microbiology for decades, but are a relatively new topic in electronics design and research. Most low-power electronics have traditionally opted for batteries, RF energy, or solar cells. This is changing, however, as the limitations and costs of these energy sources hamper our ability to deploy useful systems that last for decades in challenging environments. If large-scale, long-term applications like underground infrastructure monitoring, smart farming, and sensing for conservation are to be possible, we must rethink the energy source.

Building health workforce capacity in Northern Australia

The Menzies Ramaciotti Regional and Remote Health Sciences Training Centre (Menzies-Ramaciotti Centre) is located within the Menzies School of Health Research (Menzies) in Darwin, Northern Territory (NT). The Menzies-Ramaciotti Centre is contributing to the development of a local health workforce in the NT, including a strong biomedical workforce. The Centre facilitates health workforce career progression for regional and remote youth, with a focus on career development for Aboriginal and Torres Strait Islander (First Nations) youth. The Centre works in collaboration with a range of industry and education partners, who also have strong workforce development goals and a commitment to serving a vital community need to build pathways into work and study with First Nations peoples. Part of the Centre’s focus entails delivery of high-quality training in biomedical sciences, including theoretical and practical skill development in microbiology, laboratory techniques, immunology, public health, data science, allied health, and health research. The Centre uses a non-linear, strengths-based approach to training with a multiplicity of entry and exit points including high school work experience placements, traineeships, vocational placements, as well as undergraduate and postgraduate placements.

Oral Microbiome and Dental Caries Development.

Dental caries remains the most prevalent oral disease worldwide. The development of dental caries is highly associated with the microbiota in the oral cavity. Microbiological research of dental caries has been conducted for over a century, with conventional culture-based methods and targeted molecular methods being used in order to identify the microorganisms related to dental caries. These methods’ major limitation is that they can identify only part of the culturable microorganisms in the oral cavity. Introducing sequencing-based technology and bioinformatics analysis has boosted oral microbiome research and greatly expanded the understanding of complex oral microbiology. With the continuing revolution of molecular technologies and the accumulated sequence data of the oral microbiome, researchers have realized that microbial composition alone may be insufficient to uncover the relationship between caries and the microbiome. Most updated evidence has coupled metagenomics with transcriptomics and metabolomics techniques in order to comprehensively understand the microbial contribution to dental caries. Therefore, the objective of this article is to give an overview of the research of the oral microbiome and the development of dental caries. This article reviews the classical concepts of the microbiological aspect of dental caries and updates the knowledge of caries microbiology with the results of current studies on the oral microbiome. This paper also provides an update on the caries etiological theory, the microorganisms related to caries development, and the shifts in the microbiome in dental caries development.

ISOLATION AND CHARACTERISTICS OF BACTERIA CITROBACTER SP. SR35 FROM A COAL MINE WASTE DUMP

SummaryBacteria of the genus Citrobacter are found in soil, water, the intestinal tract of animals, human clinical samples (urine, sputum, blood, wound drainage, etc.), also in wastewaters and waste dumps of mines. Gram-negative bacteria SR35, capable of reducing sulfur and sulfate ions, were isolated from the waste dump of the Nadiya coal mine, Chervonograd Mining and Industrial district (Lviv region, Ukraine). The work aimed to identify and study morphological, physical and chemical properties (cell shape, size, Gram staining, sporulation, motility, oxygen requirements, ability to form H2S, utilization of carbon sources, catalase activity, oxidase activity) of isolate SR35. Methods. We used standard microbiological and biochemical research methods (microbial culture, microscopy methods, determination of amilase, lipase activity). Chromosomal DNA was isolated by the method of soft lysis. The 16S rRNA gene was amplified using universal primers 27F and 1492R. It was sequenced by the Sanger method. Phylogenetic reconstruction was performed using the MEGA X program. Identification of isolates was carried out based on both the sequence of the 16S rRNA gene and physiological and biochemical properties. Results. The studied bacteria are rods (0.5–0.8×1.5–2.0μm), which form chains and are capable of reducing nitrate and sulfate ions and assimilating several carbon sources. Glucose, lactose, mannose, mannitol, and inositol are fermented with the formation of acid. Motile. Catalase positive, oxidase negative. Form H2S during growth in Kligler’s medium. According to the results of pairwise alignment of the isolate’s 16S rRNA gene sequence, the highest percentage of identity with representatives of the genus Citrobacter was established (99.23–99.86% identity, coverage 98%) and it was confirmed by phylogenetic reconstruction. Citrobacter sp. SR35 is resistant to 2 μM cadmium (II), 5 mM iron (II), 0.25 mM cobalt (II), 10 mM manganese (II), 0.5 mM copper (II), and 0.1 mM chromium (VI). Conclusions. According to the results of sequencing of the 16S rRNA (accession number GenBank OP279754) gene, and physiological, and biochemical characteristics (oxidase, catalase, metabolism of carbon sources, production of H2S, etc.), it was established that isolate SR35 belongs to the genus Citrobacter.

Assessing decontamination practices at a medical microbiology research laboratory

To our knowledge, this is the first study to conduct an objective assessment of the routine decontamination practices at a medical microbiology research laboratory (MRL) a year after a biosafety training was provided to all laboratory staff. Between March 28th and June 28th, 2021, unobtrusive observations were carried out to identify-three high-touch surfaces at the MRL during different working hours. Swabbing was used to evaluate the effectiveness of the disinfectant used in the laboratory. All three high-touch surfaces were sampled before and after decontamination with 200 ppm of 5 % sodium hypochlorite (household bleach) to quantify the microbial load and identify the types of organisms residing on the laboratory surfaces. A higher concentration (500 ppm) of 5 % sodium hypochlorite was employed after refresher training was provided to housekeeping staff, and resampling of the three surfaces was carried out during a 4-week follow-up period using the same procedure. The three high-touch surfaces identified were the two sides of the workbench (22 %–24 %) and the front surface of one incubator (14 %). Anthracoid bacilli and Staphylococcus aureus were the most commonly found organisms on laboratory surfaces pre-intervention (100 % and 89 %, respectively) and post-intervention (56 % and 44 %, respectively). Other microorganisms detected included Salmonella spp. (27.7 %), Proteus spp. (5.6 %), Escherichia coli (5.6 %), and Klebsiella spp. (33.3 %). Employing a higher concentration (500 ppm) of sodium hypochlorite significantly (p ≤ 0.000) reduced the total aerobic colony count from an average of 15–250 cfu/cm2 to 10–60 cfu/cm2. This study demonstrated suboptimal decontamination practices at the MRL and the need to apply a higher concentration (500 ppm) of sodium hypochlorite to reduce the overall microbial load. It also demonstrated the importance of quantitative assessment to monitor decontamination practices and ensure staff compliance. More studies are needed to identify bacterial communities within the laboratory, which will help provide guidance regarding the types, proper concentrations, and appropriateness of the in -use disinfectants. Furthermore, large-scale studies on the acceptable level of residual contamination following any decontamination process are urgently recommended.

Prevalence of Virulence Genes Among Escherichia Coli Strains Isolated from Food and Carcass Swabs of Different Animal Origins in Croatia.

Escherichia coli is present in the normal intestinal flora but some strains can cause intestinal and extraintestinal diseases, and research on its presence in food of animal origin is in the interests of public health. This study was designed to characterise E. coli strains according to their origin, their carriage of virulence genes specific for certain pathogroups, and phylogenetic group affiliation. The study was carried out on 100 E. coli strains isolated from food samples of various animal origin as well as pig and cattle carcass swabs. Isolation of the strains was performed using two methods. One method included colony count and the other an overnight enrichment of the samples. Isolation was followed by DNA extraction and detection of virulence genes and phylogenetic group with conventional and multiplex PCRs. In this study, the most prevalent gene was EAST1 (20%) and strains which carried it were identified as enteroadherent E. coli. Other pathogroups were represented in lower incidences. Phylogenetic group analysis revealed the prevalence of the A and B1 groups, with B1 mainly present in game and cattle strains, while the majority of pig and poultry strains were assigned to group A. This study provides an overview of the presence of potentially pathogenic strains and E. coli phylogenetic groups in Croatia, for which the data are limited. Further microbiological and molecular research is required to examine the epidemiological situation in the country.

The Influence of Chestnut Flour on the Quality of Gluten-Free Bread

Gluten-free bread is the basis of an elimination diet in the case of many glucose-related diseases. The quality of this bread differs significantly from traditional products; therefore, it is necessary to conduct research aimed at improving the quality of this type of product. The aim of the study was to determine the effect of the addition of chestnut flour and the method of packaging on the quality of gluten-free bread. The addition of chestnut flour (partially replacing corn starch) was used in the amount of 5, 10, 15 and 20% of the total weight of the concentrate. The influence of the storage method on the quality of the tested bread was examined after 7, 14 and 21 days from baking. The refrigerated breads were packed using PA/PE barrier foil with air and vacuum (58%) and were stored in room temperature (22 ± 2 °C). Water content, texture and color were determined, and sensory evaluation and microbiological analysis were performed. As a result of the conducted research, we observed that the addition of chestnut flour to the recipe affects significantly (p < 0.05) the texture of the finished product, reducing the hardness and increasing the elasticity and cohesiveness of the bread crumb. The use of chestnut flour in an amount of up to 10% increases significantly (p < 0.05) the volume of the resulting loaves. Microbiological research has indicated vacuum packaging as a better way to protect and store gluten-free bread. For practical use in future production, it is recommended to replace corn starch in gluten-free breads by no more than 10% by chestnut flour.

Development and Use of a Kinetical and Real-Time Monitoring System to Analyze the Replication of Hepatitis C Virus.

In microbiological research, it is important to understand the time course of each step in a pathogen’s lifecycle and changes in the host cell environment induced by infection. This study is the first to develop a real-time monitoring system that kinetically detects luminescence reporter activity over time without sampling cells or culture supernatants for analyzing the virus replication. Subgenomic replicon experiments with hepatitis C virus (HCV) showed that transient translation and genome replication can be detected separately, with the first peak of translation observed at 3–4 h and replication beginning around 20 h after viral RNA introduction into cells. From the bioluminescence data set measured every 30 min (48 measurements per day), the initial rates of translation and replication were calculated, and their capacity levels were expressed as the sums of the measured signals in each process, which correspond to the areas on the kinetics graphs. The comparison of various HuH-7-derived cell lines showed that the bioluminescence profile differs among cell lines, suggesting that both translation and replication capacities potentially influence differences in HCV susceptibility. The effects of RNA mutations within the 5′ UTR of the replicon on viral translation and replication were further analyzed in the system developed, confirming that mutations to the miR-122 binding sites primarily reduce replication activity rather than translation. The newly developed real-time monitoring system should be applied to the studies of various viruses and contribute to the analysis of transitions and progression of each process of their life cycle.