The half-life of cyanidin-3-glucoside in small hydrogel beads was roughly twice as long as that of the larger beads under accelerated light exposure at 4 °C and they were less likely to undergo noticeable color changes over time. Both sizes of hydrogel beads stored at 97% RH and 4 °C showed color stability over the 21-day period with minimal color variation (|ΔE| ≤ 3). The UV–vis spectra of the purple corn extract exhibited changes across pH 2 to 12, as evidenced by the visible color variations, ranging from pink to green. The limit of detection (LOD) for NH3 was 25 ppm for small beads and 15 ppm for large ones. Both types of beads exhibited similar LOD for DMA and TMA, around 48 ppm. This research showed that alginate hydrogel beads containing anthocyanins from purple corn are a viable option for developing intelligent packaging of muscle foods. Furthermore, the use of hydrogel beads of different sizes can be customized to specific muscle foods based on the primary spoilage compound generated during spoilage.pH indicators have emerged as promising tools for real-time monitoring of product freshness and quality in intelligent food packaging applications. However, ensuring the stability of these indicators is critical for practical use. This study aims to evaluate the stability of anthocyanins-loaded alginate hydrogel beads of varying sizes at different temperatures under accelerated light conditions and relative humidity (RH) levels of 53% and 97% during 21 days of storage. Moreover, their sensitivity to the principal spoilage volatiles of muscle food products such as ammonia (NH  •Anthocyanins-loaded alginate hydrogel beads were introduced as freshness indicators.•Anthocyanins were loaded into alginate hydrogel beads of different sizes.•Color stability assessed under light and simulated relative humidity of fresh muscle foods.•Developed hydrogel-based indicators were sensitive to ammonia, dimethylamine, and trimethylamine.•Small and large beads were suitable for integration into specific muscle foods. Disposiin-situ information about the physical, chemical, and microbiological quality to consumers, retailers, and manufacturers, thereby mitigating food waste  and a heterocyclic ring (C) . One of Zea mays L.) has been extensively used as a commercial source of anthocyanins and is categorized by the European Union with code E−163 (iv) as a food colorant (purple corn extract (PCE) are anthocyanins, commonly cyanidin-3-glucoside, cyanidin-3-, pelargonidin-3-glucoside, pelargonidin-3-, peonidin-3-glucoside and peonidin-3- (Purple corn (colorant . The maiucoside) . Cyanidiucoside) . Furtherucoside) . Acylateucoside) .2+)  .Even though solvent casting and coating a filter paper or polymeric films with a dye solution are frequently used in preparing pH-indicator films, pH-indicators produced with these techniques must be sufficiently sensitive and fast-responding in most cases. Encapsulating the anthocyanins in alginate beads with electrostatic extrusion provides a high level of versatility in terms of particle size and morphology, depending on operating parameters . Alginat3), dimethylamine 2N), and trimethylamine 3N) are the significant components produced during muscle food spoilage because of microbial activities. These volatile amines are also referred to as total volatile basic nitrogen (TVBN), which is used to measure the freshness of the seafood , ATP degradation products, and detection of volatiles in the headspace with GC-MS are routinely performed. Despite the robustness of these techniques, they are time-consuming, destructive, costly, and require specific equipment and specialists. Therefore, using colorimetric indicators to evaluate seafood freshness and facilitate the decision-making process may be promising  for TMA and DMA was investigated in the current research, thereby contributing to more accurate assessment of freshness in muscle food products.22.1Laminaria hyperborea, Mw= 2.74×105 g mol−1, Protanal LF 200S) was obtained from DuPont Nutrition and Health . Cyanidin-3-glucoside (C3G) was purchased from PhytoLab GmbH & Co KG . Other chemicals were provided by Sigma Aldrich.Sodium alginate -propionic-acid sodium salt in D2O  was added for internal chemical shift reference. 1H NMR spectrum was obtained by BRUKER NEO 600 MHz equipped with 5 mm iProbe TBO  and recorded at 83 °C. The spectra were recorded using TopSpin 4.0.7 software (Bruker BioSpin) and processed and analyzed with TopSpin 4.0.7 software (Bruker BioSpin). The chemical composition of alginate and NMR spectrum is shown in The chemical composition of the alginate was analyzed using NMR spectroscopy, according to the standard test method established by the American Society for Testing and Materials  and prev2.3g at 4 °C for 10 min. The final aqueous extract was obtained by filtration using a syringe filter .Purple corn powder was purchased from Dragon superfoods, Bulgaria. Extraction of anthocyanins was performed according to 2.42.4.1Cyanidin-3-glucoside has been identified as the predominant anthocyanin in PCE . PCE was2.4.2The pH sensitivity of PCE was investigated by mixing 700 μL PCE in 5 mL of 10 mM tris buffer solutions (pH 2–12). After 20 min, the UV–Vis spectra (450–700 nm) of the obtained solutions were recorded by a UV–Vis spectrophotometer .L0*, a0*, and b0* are lightness, red–green, and yellow-blue color attributes of diluted PCE, while L*, a*, and b* are the color parameters after buffer addition.Color parameters were determined via a DigiEye Imaging system , consisting of a cabinet adjusted to match the D65 illuminant and a digital camera , centrally mounted on top of it. A Digi Eye Digitizer Chart was used to calibrate the system and relate the camera's RGB signals to the CIE specifications. Images were analyzed with DigiPix Software Version 2.9. Total color difference (ΔE) was calculated using the following formula as described by CIE International Commission on L0*, a0*, and b0* are the color parameters of diluted extracts.To ensure an accurate comparison and evaluate the influence of pH and dilution on the color parameters of the extracts, control extracts were prepared by diluting with a buffer having the same initial pH as the extracts (5.2 for 26% PCE and 5.6 for 20% PCE). To this end, 5 mL of tris buffer solution with pH values of 5.2 and 5.6 was added to 700 μL of 26% and 20% extracts, respectively. Therefore, 2.52 and 0.1 M HCl in the extract (pH= 2.5). The operating voltage was 5 kV for the 20% extract and 3 kV for the 26% extract. The distance from the nozzle tip to the cross-linking solution was set to 5 cm. After droplet penetration, the beads were left to harden for 30 min to ensure thorough gelation. The beads were then filtered and rinsed with deionized water and 70% ethanol to remove impurities. Anthocyanins were encapsulated within alginate beads using a custom-made electrostatic droplet generator , based on the optimal conditions reported in our previous study . Sodium 2.62.6.1w) was analyzed using Novasina aw meter . The measurements were performed in triplicate.The moisture content of the beads was determined gravimetrically by evenly spreading 1 g of the beads in petri dishes. The samples were dried at 70 °C to a constant mass . The bea2.6.2The size of the hydrogel beads was determined through an optical light microscope (Nikon Eclipse TS100). Images of the particles were captured, and the diameter of 100 beads was measured.2.6.3g for 20 min at 4 °C, and the obtained supernatant was filtered using a 25 mm, 0.45 μm syringe filter made of Polyethersulfone (Sigma Aldrich). The C3G content was determined using the HPLC method outlined in section Encapsulation efficiency (EE) was quantified using the method described by 2.72.7.12> 0.9)  of C3G during storage was calculated as follows:The color stability and degradation kinetic of C3G in hydrogel beads was evaluated under light (11700 ±100 Lux) at ambient (23 °C) and chilled storage temperatures of commercial food products (4 °C). Random samples were collected at specific time intervals  and color parameters (CIE Lab) were determined with DigiEye imaging system as described in section 2> 0.9) . A plot 2.7.2w, and color properties were determined on days 0, 1,3,5, 7, 14, and 21. The experiment was carried out in triplicate.The stability test under different relative humidity (RH) levels was assessed at RT and 4 °C. Desiccators with two relative humidity levels, 53%, and 97% were obtained by saturated magnesium nitrate and potassium sulfate solutions, respectively. RH was monitored and verified in each chamber at equilibration with a hygrometer . Hydrogel beads were placed in each desiccator and kept in the dark. C3G content, a2.8Hydrogel beads were investigated for their ability to discriminate three volatile amines. To assess the color response of the fabricated indicators to ammonia, TMA, and DMA, beads were placed on a permeable fabric and attached to the mouth of Scotch bottle containing 25 mL various concentrations of ammonia , DMA and TMA . The exposure was done for 24 h, and colorimetric values were recorded. The limit of detection (LOD) and limit of quantification (LOQ) were calculated based on the total color difference versus volatile concentration (ppm) in the linear range from the formulas below :(Eq.4)LO2.9t-test.Each experiment was performed in triplicate, and the results were reported as the mean value with the standard deviation (SD). In the stability studies, temperature and storage time were considered the primary factors and analyzed through the general linear model (GLM). One-way analysis of variance (ANOVA) was carried out to evaluate the sensitivity to volatiles and colorimetric change at various pH levels. The difference was considered statistically significant if p < 0.05, and the post hoc Tukey's test was conducted using SPSS software (version 28.0.1.0 (142)). Two anthocyanins-loaded hydrogels were compared by independent sample 33.1mg/100 mL and 6.33±0.1 mg/100 mL in 20% and 26% extract, respectively. The color change of PCE was examined under various buffer solutions to confirm its suitability as a pH indicator. 1. Under mild acidic conditions, flavylium cation undergoes a competitive process, including hydration and deprotonation, leading to hemiketal and neutral quinonoid base formation as depicted in cis) and trans-chalcone (Ctrans), which exhibit a yellow color. The neutral quinonoid bases, on the other hand, transform into anionic quinonoid bases as the pH increases towards neutrality via losing a second proton  in color values between the two concentrations. As expected, the 20% PCE exhibited lighter color (L*= 67.48±0.08) with lower redness (a*=13.58±1.18) and yellowness (b*=4.15±0.02) values than the 26% PCE . The color properties  of the two extracts at pH solutions ranging from 2 to 12 are depicted in 3.21 and F2 hydrogel beads was over 96% and 93%, respectively, with the latter showing higher solids content . The retention of C3G in both hydrogels decreased to approximately 10% of the initial concentration at RT on day 14. It continued to fall until nearly zero at the end of the 21 days  of C3G in F1 and F2 subjected to light and under dark conditions. The results displayed that lowering the storage temperature led to a significant increase in the half-life of the C3G, as well as a reduction in the degradation rate constant (k) (p< 0.05). Furthermore, it was observed that the light-exposed samples were less stable compared to samples stored in the dark. The half-life of C3G in F1 hydrogel exposed to light at refrigerated conditions was shorter, with a value of 15.9 days, compared to the same samples stored in the dark, where the half-life was estimated to be approximately 10 times longer, 157 days. Similarly, for F2, the half-life was 8.43 days when exposed to light and 81.8 days when stored in the dark. The photochemical degradation mechanism of anthocyanins is a complex process that involves various reactions and intermediates. When anthocyanins are exposed to light, they absorb energy and are excited to a higher energy state. This excited state can lead to several different reactions that result in the degradation of the molecule. One proposed mechanism involves the formation of a C4 adduct intermediate, which is formed by the hydrolysis of position C4 in the flavylium excited state. This intermediate can then undergo subsequent hydrolysis and ring opening to create various degradation products, including aromatic aldehydes, carboxylic acids, and phenolic compounds. Another suggested mechanism involves the hemolytic scission of the C2–C3 bond of the chalcone form, which can lead to the formation of various degradation products. This reaction is postulated to occur via a radical pathway, which involves the formation of a radical intermediate that can undergo multiple subsequent reactions  are less susceptible to noticeable color changes compared to F2 as time extended. Therefore, it is recommended to apply the F1 freshness indicator for muscle food products with both short and long shelf lives. On the other hand, the F2 indicator is most suitable for highly perishable muscle foods.The color parameters L*, a*, and b* were measured to evaluate various aspects of color, including lightness (L* 100/0 white/black), redness (+a*), greenness (-a*), yellowness (+b*), and blueness (-b*). On day 0 of storage, the hydrogel beads exhibited L*, a*, and b* values of 54.73 ± 1.71, 37.88 ± 1.47, and 8.39 ± 1.84 for F1 B and 42.1 . However, the temperature significantly impacted the water activity only under lower RH (53%), regardless of the type of beads investigated. A steady trend with negligible fluctuations in aw was observed at a 97% RH in all studied temperatures. Anthocyanins-loaded alginate hydrogel beads stored at 53% RH underwent moisture loss to achieve equilibrium with the surrounding environment. F1 showed a rapid reduction in moisture in comparison to F2, which is due to their higher surface-to-volume ratio and smaller particle size , hydrogel beads containing anthocyanins tend to dry out, shrink, and darken, which makes them less suitable for use. Therefore, the focus of the study was to investigate the retention of C3G under simulated condition of 97% relative humidity.The retention of C3G in both hydrogels was significantly influenced by temperature and storage time, with a notable interaction effect between the two factors (p< 0.05). As the temperature increased, the retention of C3G significantly decreased (p< 0.05), leading to approximately 40% greater C3G loss by the end of day 21 at RT compared to storage at 4 °C for the same samples. Anthocyanin degradation under current storage conditions is likely related to the cleavage of covalent bonds, which form colorless compounds as temperature increases. It can be postulated that the water molecules in the hydrogel structure accelerates the rate of hydrolysis and break down the glycosidic bonds, resulting in the loss of red color . Apart f1 and F2 significantly decreased (p< 0.05) over the storage period, with F1 showing better retention than F2 at both stored temperatures. At refrigerated temperature, the concentration of C3G in F1 significantly declined after 7 days, reaching 80% of the initial concentration, while F2 showed a significant reduction after 5 days, with 79% of the initial concentration remaining  provided better protection. Instead, pore volume and pore size appear to play a critical role in retention of C3G as larger pores facilitate water diffusion and promote interactions with anthocyanins. The pore volume and pore size of F1 were 0.08 cm3/g and 17.4 nm, respectively, while F2 had a pore volume of 0.44 and a pore size of 34.68 nm  produced by microbial activities during the spoilage of protein-rich food products. Thus, the relatively high sensitivity of hydrogel beads to the targeted analytes might reflect the indicator's potential to respond to pH changes.The pH of the packaging headspace is influenced by volatile metabolites . NH4+ creates an alkaline environment on the surface of the bead. This causes the phenolic hydroxyl group to undergo an acid-base reaction with OH− and, ultimately, the formation of a phenolic oxygen anion in the anthocyanin structure , whereas an increase of 127% was recorded in F1 in higher concentrations, between 12 and 21 ppm. ΔE values of F1 and F2 reached their maximum at 300 ppm, corresponding to 34 and 31, respectively.Anthocyanin-loaded alginate hydrogel beads were investigated for their responsiveness to ammonia by placing them on a permeable fabric attached to the mouth of a bottle. The beads were exposed to different concentrations of ammonia ranging from 1 to 300 ppm for 24 h, and color values were recorded. tructure . Lightneo 12 ppm A. Then b3 concentrations was observed between 0 and 12 ppm. F2 was found to be more sensitive than F1 to ammonia. LOD values were reported to be 25 and 15 ppm for F1 and F2, respectively. Meanwhile, LOQ values were 83 and 50 ppm. R2 values above 0.9 indicate the calibration curve's high detection sensitivity and precision , anthocyanins content, initial color, and degree of lightness. Based on the morphological observation in our earlier study  is reached, lighter-colored hydrogel beads become more effective at detecting color changes. This implies that the smaller the diameter of the beads, the lighter they appear, resulting in a more efficient detection, particularly at higher concentrations.The limit of detection (LOD) and limit of quantification (LOQ) refer to the lowest concentration of an analyte that can be calculated reliably through a specific method of measurement . These drecision . Severaler study , the F2 r change . Structu1 and F2 indicators, in turn, were at least 8 and 13-times more sensitive to NH3 than cellulose fiber embedded with red cabbage anthocyanins , considering stability. The use of hydrogel beads in the packaging of products with low relative humidity was found to be unsuitable due to issues such as excessive moisture loss and a darkened appearance. Thus, freeze-drying of hydrogels can be a potential alternative for future studies. The sensitivity of the hydrogel beads to specific amines varied. Large beads exhibited higher sensitivity to lower ammonia concentrations, making them well-suited for early-stage spoilage detection or products with lower levels of ammonia production. On the other hand, small beads were appropriate for products with higher levels of ammonia production. Both hydrogel beads showed similar limits of detection for dimethylamine and trimethylamine. However, significantly different performance was observed at concentrations exceeding 1000 ppm in a way that the small and large beads were more sensitive to trimethylamine and dimethylamine, respectively.The results of this research provide significant insights into the practical application of these hydrogel bead indicators in different types of muscle food and highlight the importance of tailoring the choice of hydrogel bead based on specific product characteristics and the desired stage of spoilage detection. Overall, hydrogel beads developed in this study successfully exhibited desirable properties, such as color stability and responsiveness to volatile amines. However, the stability of anthocyanins plays a pivotal role in ensuring the long-term functionality of a pH indicator, and there is still room for further stability improvement through using fillers, antioxidants, or protective layers.Samira Mohammadalinejhad: Conceptualization, Methodology, Formal analysis, Investigation, Visualization, Writing – original draft. Marcin Kurek: Supervision, Writing – review & editing. Ida-Johanne Jensen: Supervision, Writing – review & editing. Jørgen Lerfall: Conceptualization, Methodology, Supervision, Project administration, Writing – review & editing.I confirm that there are no known conflicts of interest associated with this manuscript and there has been no financial support for this work that could have influenced its outcome. I confirm that the manuscript has been read and approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed.


This deceptively simple question remains challenging to address today, yet because it relates to fundamental processes and characteristics of viral communities, even the imperfect approximations available have been critical for the field of viral ecology. Most notably, early quantitative observations reporting a high abundance of virus-like particles in aquatic environments , 2 put aThis “virus to cell” ratio, also referred to as the virus-to-microbe ratio (VMR or VTM), virus-to-bacterium ratio (VBR), or virus-to-prokaryote ratio (VPR), is often reported as a key ecological metric, and typically derives from counts of viruses and cells using transmission electron microscopy (TEM), epifluorescence microscopy (EFM) or flow cytometry (FCM). Among these, EFM counts have been the most widely used as they combine a relatively high throughput and an ability to distinguish even small viruses from background noise . These aIn aquatic environments, a large-scale meta-analysis indicated that, while a 10:1 ratio could represent a median value for some ecosystems, VMRs typically span across ~ 2 to 3 orders of magnitude so that “a 10:1 model has either limited or no explanatory power” . A similMeanwhile, these last 15 years have seen a rapid transformation of the viral ecology field with the fast rise of metagenomics. Large genome catalogs for uncultivated viruses obtained from metagenomes already provided invaluable information regarding their functional diversity, distribution across ecosystems, and dynamics through space and time . Given tIn a new study, López-García et al. try to tackle this challenge by formally establishing a metagenome-based virus to (cellular) microbe ratio (mVMR), and compare it to microscopy-based counts across several ecosystems and sample types including bulk samples, cellular fractions, and viral fractions . The mVMApplied to a broader range of samples and ecosystems, mVMR typically range between 1:1 and ~10:1 with a relatively high sample-to-sample variation. Some broad ecosystem trends were nevertheless apparent, with higher mVMR in aquatic ecosystems compared to soil/sediments and animal-associated microbiomes. López-García et al. also illustrate another potential advantage of the mVMR approach by providing an estimated relative abundance of individual taxa within the aggregated “virus” and “microbe” counts for each sample, leveraging the fact that single-copy genes can also be used as taxonomic markers. This larger analysis illustrates both the promises and the remaining challenges of mVMR as a viral ecology metric. On one side, the prospect of leveraging the ever-growing number of public metagenomes, along with the possibility to estimate mVMR in a lineage-specific way to more closely reflect virus:host relationships, are tantalizing and would enable a much larger and in-depth investigation of virus:host dynamics across microbiomes. On the other hand, it is still complicated at this point to determine whether mVMR applied in this way is a “superior” metric, i.e., more accurate and less biased than microscopy counts.Several technical limitations could lead to both under- and over-estimation of mVMR. Publicly available metagenomes most often target cellular size fractions, and may miss a significant number of viruses including (i) viral genomes encapsidated in virions, (ii) viruses not represented in current collections of single-copy marker genes, and (iii) ssDNA and RNA viruses not captured by standard library preparation protocols. While the most common dsDNA viruses are now most likely well captured by single-copy marker genes, a comprehensive mVMR would have to rely on integrated sampling across size fractions and a combination of DNA and RNA libraries, which would not be available for the majority of publicly available data. Meanwhile, mVMR counts will also include elements not typically considered in the VMR metric, e.g., dormant viruses that do not produce virions and/or kill their host, or virus-like machinery encoded by microbes and including homologs of virus marker genes. Hence, with the potential for relatively large under- and over-estimation, it is still difficult to get a sense for how accurate the mVMR metric is.Ultimately, mVMR and fVMR appear to provide orthogonal perspectives on a highly complex biological process with different technical biases and limitations. On a bad day, a viral ecologist would only see two flawed metrics with unknown intervals of confidence that cannot be trusted. But on a good day, these two approaches can easily be seen as highly complementary, and the addition of mVMR next to fVMR could provide a much-needed additional window into virus:host interactions in microbiomes. With more controlled studies of known consortia to better evaluate methodological biases, advances in “quantitative sequencing” with the use of e.g., artificial spike-ins or high-throughput single-cell approaches, and more studies providing paired microscopy- and sequencing-based observations, we believe that the limitations around fVMR and mVMR will progressively be reduced, finally enabling viral ecologists to be more confident in these measurements. For better or worse, however, since both fVMR and mVMR seem to suggest that viruses typically outnumber cells across diverse samples and microbiomes, we anticipate to keep reading variants of the “viruses are the most abundant …” introduction statement for the foreseeable future.


Surveillance of viral pathogens in both point-of-care and clinical settings is imperative to preventing the widespread propagation of disease—undetected viral outbreaks can pose dire health risks on a large scale. Thus, portable, accessible, and reliable biosensors are necessary for proactive measures. Polymeric microparticles have recently gained popularity for their size, surface area, and versatility, which make them ideal biosensing tools. This review cataloged recent investigations on polymeric microparticle-based detection platforms across eight virus families. These microparticles were used as labels for detection (often with fluorescent microparticles) and for capturing viruses for isolation or purification (often with magnetic microparticles). We also categorized all methods by the characteristics, materials, conjugated receptors, and size of microparticles. Current approaches were compared, addressing strengths and weaknesses in the context of virus detection. In-depth analyses were conducted for each virus family, categorizing whether the polymeric microparticles were used as labels, for capturing, or both. We also summarized the types of receptors conjugated to polymeric microparticles for each virus family. Viruses significantly burden public health, and viral outbreaks have increased in recent years . WithoutViruses have typically been detected with amplification-based methods, including polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR), which are reliable methods for obtaining highly-specific results . The fleNon-amplification-based methods present alternative solutions to monitoring viral outbreaks. These include serological tests like hemagglutination-inhibition assays (HIA), immunofluorescence assays, or chemiluminescent immunoassays (CIA). Serological tests typically employ a functionalized system to detect the presence of viral antigens or antibodies for a viral target. They are much simpler, low-cost, and provide much shorter assay time (<15 min) than amplification-based methods. However, less-than-perfect specificity caused by antibody cross-reactivity increases the risk of false-positive results . TherefoNevertheless, particle-based modifications to serological tests have been proposed, with results demonstrating high specificity and sensitivity ,16. SpecMicroparticles have also been used for numerous serological tests in the past, many of which are polymers, e.g., polymeric microspheres, latex beads, etc. Microparticles present unique ways to modify amplification-based and non-amplification-based viral detection methods. Microparticles can easily be removed from the solution through centrifugation or applying a magnetic field; they can also aid in simplifying and shortening procedures . In addimicrospheres or microbeads. Fabrication methods for microparticles vary depending on the size and material of the particle. Many microparticles for virus detection are polymeric microparticles, often called latex particles or latex beads. Most polymeric microparticles are polystyrene (PS), or co-polymers of PS and other polymers, typically synthesized via emulsion polymerization. Many PS-based microparticles are now commercially available, from companies including MagSphere, Inc. and Bangs Laboratories, Inc. Their sizes range from 0.1 μm to 10 μm, with surface modified types including carboxylated, aminated, sulfonated, etc. . These commercial PS-based microparticles can also incorporate fluorescent dyes or magnetic properties.Microparticles are spherical particles between 1 and 1000 µm (sometimes 0.1 and 1000 µm) in diameter . These cMore sophisticated microparticle fabrication technologies have recently been introduced towards more uniform size and improved surface properties. For example, microparticles can be synthesized within a microfluidic device, where emulsion droplets are converted to solid particles through a chemical or photo-polymerization technique . TemplatDue to their size properties, microparticles are ideal for centrifugal separation techniques and can be imaged via conventional microscopy, enhancing the simplicity and accessibility of procedures involving microparticles. Lastly, microparticles have been engineered for drug delivery applications, which this review will not cover as it is primarily concerned with virus detection. submicron particles is sometimes used for these particles (100 nm–1 μm). In this review, however, we will consider submicron particles as a part of microparticles.Although similar to microparticles, nanoparticles are primarily distinguished from microparticles in size and electrochemical properties. Nanoparticles are often defined with a diameter of less than 100 nm, and they can be made from the same materials which constitute microparticles, most notably metals and polymers . Due to Both microparticles and nanoparticles have been employed for viral sensing applications. This review, however, will limit the discussion to using microparticles for virus detection.Microparticles have previously been used for serological tests, targeting either virus antigens or antibodies generated from immune responses. Typically, antibodies are conjugated to microparticles to detect virus antigens in blood sera, and antigens are conjugated to microparticles to detect neutralizing antibodies in blood sera. The same methods can be duplicated for saliva tests, which have been popularly used during the SARS-CoV-2 pandemic. Such methods can also be used for screening virus antigens in environmental samples, such as aerosols, field water samples, and food samples. However, the number of such virus antigens in environmental samples tends to be very low, requiring very low LODs, which are often difficult to achieve in traditional immunoassays .Enzyme-linked immunosorbent assay (ELISA) is one of the oldest immunoassay methods, but it is still popular . For detMicroparticles are recently gaining popularity in numerous immunoassay platforms, including microplates, LFIAs, and microfluidic chips. They can replace the role of enzyme–substrate pairs, AuNPs, or fluorescent dyes. The substantial size of microparticles may allow them to generate optical signals strong enough for the naked eye to recognize. In addition, antibody-conjugated microparticles can be aggregated together via antibody–antigen binding, known as a particle immunoagglutination assay d. In thiamplicons) from a sample and be physically separated under an external magnetic field Label: microparticles are used as labels for virus detection;(2)Capture: microparticles are used for separating, purifying, capturing, or extracting virus antigens and amplicons from samples;(3)Both: label and capture;(4)Other: microparticles are used for other purposes.The use of microparticles for virus detection can be classified into four categories:(1)Antibody ;(2)Antigen ;(3)Aptamer ;(4)Nucleic acid (mainly to capture amplicons);(5)Other.For all applications, microparticles must be conjugated with receptors. Based on the receptor type, they can be classified into five categories:In the following sections, we will summarize the use of (polymeric) microparticles for each virus family based on their use  and the receptor type .While any virus can be detected using microparticles on various detection platforms, we summarized a list of viruses that have been detected using microparticles. Viruses are infectious microbes ranging in size from 0.02 to 0.3 µm, composed of genomic DNA or RNA and a nucleic protein capsid . These pCoronaviridae, Filoviridae, Flaviviridae, Hepadnaviridae, Herpesviridae, Orthomyxoviridae, Papillomaviridae, and Retroviridae.The five primary transmission routes for viruses include direct contact, aerosols, fomites, ingestion, and vectors . Given tWe searched journal articles for microparticle-based virus detection from the beginning of 2018 to the end of June 2023, i.e., 5.5 years, to focus on the recent trends. The first 2 years represented the pre-pandemic era, while the remaining 3.5 years fell into the pandemic and post-pandemic periods. The target viruses were classified into eight families, as shown below. We could find almost no articles on two virus families—filoviruses and hepadnaviruses—and we had to search further back, to 2015, for these two.Coronaviridae (coronaviruses) is a family of relatively large, enveloped, positive-sense RNA viruses . I. I51]. IMagnetic microparticles can also facilitate nucleic-acid-based detection strategies for SARS-CoV-2. For example, amplified products (amplicons) of SARS-CoV-2 were captured on magnetic microparticle surfaces, inducing magnetic-field-induced agglutination of amplicon-captured microparticles. This agglutination makes a signal visible to the naked eye in as little as 20 min . AchieviFiloviridae (filoviruses) contains non-segmented negative-sense RNA . A. A62]. Ananotags. Antibody-conjugated magnetic microparticles were used to capture antigens. After mixing these reagents with a whole blood sample for only 25 min, an external magnet helped pelletize the antigens. In detection, only one excitation light could uniquely excite all three nanotags without interfering with the natural absorbance wavelength of whole blood, effectively eliminating the need for sample preparation. Magnetic microparticles were also used to separate or capture EBOV targets, followed by other detection methods. Xu et al. demonstrated a simple EBOV DNA immunoassay with electro-thermal atomic absorption spectrometry . A simplCoxiella burnetii, African swine fever virus (ASFV), West Nile fever virus (WNV), Lyme disease pathogen Borrelia burgdorferi, vesicular stomatitis virus (VSV), Rift Valley fever virus (RVFV), Ebola virus (EBOV) and Schmallenberg virus (SBV) . S. S82]. SPolymerized QDs were also used for multiplexed detection of HBV. When used in tandem, the potent fluorescence of QDs and the target agnostic surface functionalization of polymeric microparticles could generate an individualized signal for single targets. In this manner, multiple targets could be detected from one solution. Using this method, Kim et al. attained a clinical sensitivity range of 80.4–90.5% for multiplexed HBV genome detection . The othThe herpesviridae (herpes virus) family are enveloped, double-stranded DNA pathogens with extracellular glycoproteins for host transduction . HerpesvHerpes simplex virus (HSV) is a double-stranded DNA virus categorized further into HSV-1 and HSV-2. HSV infection is lifelong, with outcomes ranging from asymptomatic to life-threatening . FollowiAmong the microparticle-based biosensors in our literature survey, the herpesviruses accounted for the smallest number. Furthermore, only one of the three articles featured singleplex herpes virus detection. This suggests the need for further investigation into microparticle biosensing of the herpes virus. Wu et al. proposed a deep-learning analysis of microparticle aggregation induced by HSV d 91]. M. M91]. MIn contrast, a multiplexed assay developed by Tao et al. featured HSV detection among five other virus targets . This bianti-TAGs. Limits of detection across the 14 plasmids ranged from 103 copies/reaction to 50 copies/reaction. Given the assay’s target-agnostic nature and relative specificity, this is ideal for high-throughput screening. Compared to the other multiplexed biosensors of our review, Shi et al.’s method boasted the highest number of different targets that span multiple virus families. Such highly specific multiplex platforms hold considerable potential for the next generation of microparticle-based virus detection. In yet another multiplex biosensor, 14 respiratory virus plasmids, including HSV and Epstein–Barr virus (EBV), were simultaneously detected by Shi et al. . FollowiOrthomyxoviridae is a family of RNA-based viruses, including seven genera . Four ofswine), it has since undergone recombination to become a human threat, especially with the emergence of the novel H1N1 flu in 2009. The virus is most likely transmitted through airborne droplets, but can be shared through direct contact. While most symptoms are similar to those of other influenza, a severe case of H1N1 may result in respiratory failure and subsequent death [H1N1 swine flu is a subtype of the influenza A virus and causes infection of the host’s upper, and potentially lower, respiratory tracts . While tnt death . The US Numerous microparticle-based biosensors have been developed for various influenza A viruses, while our survey indicated a relatively even distribution between label, capture, and other functions. Nearly half of the assay emphasized magnetic or thermal properties to create simple and rapid detection with minimal user intervention ,98.Two articles showed the magnetic features of microparticles. A microfluidic platform proposed by Lu et al. used electromagnetic forces on magnetic microparticles to automate an ELISA-based H1N1 immunoassay . DropletOther articles showed assay quantification. Zhang et al. developed a unique photonic (PhC) barcode multiplex immunoassay targeting H1N1, H5N1, and SARS-CoV-2 . PhC barSome detection methods significantly utilized the cross-molecular reactions between captured virus targets. Matsubara et al. used a hemagglutination inhibition assay on a glycan-functionalized microparticle to detect influenza virus . The priSmall, non-enveloped, double-stranded DNA viruses comprise the Papillomaviridae family (papillomaviruses) . They frHuman papillomavirus (HPV) is a double-stranded DNA virus causing proliferative lesions of the cutaneous and mucosal epithelium . HPV infThe number of journal articles about the microparticle-based detection of papillomaviruses is relatively low compared to other virus families cataloged in this review. This low number may warrant more thorough investigations of microparticle-based strategies. However, even among a limited pool of detection platforms, microparticles displayed various functions. Ultrasensitive and multiplexed detection of HPV DNA was achieved using PS microparticles synthesized in-house, modified with a silica coating, and doped with an organic dye . OrganicMagnetic PS microparticles can play diverse roles in non-amplification-based HPV nucleic acid detection, credited to their ability to immobilize and manipulate captured targets. Bartosik et al. captured HPV-16 and HPV-18 DNA on a magnetic PS microparticle . Target–retroviral replication [Viruses in the Retroviridae family (retroviruses) are characterized by their single-stranded RNA nucleic information dependent on DNA polymerase for their lication . Furtherlication . cDNA trlication . Human immunodeficiency virus (HIV) is an infection in which the victim’s immune system is targeted, especially CD4 lymphocytes, monocytes, and thymocytes. In the early stages of HIV infection, also known as acquired immune deficiency syndrome (AIDS), patients experience influenza-like symptoms. However, as leukocyte counts continue to drop, the patient becomes severely vulnerable to life-threatening infections ,112. DNA walker that features a spontaneous intramolecular hybridization reaction and subsequent carboxyfluorescein cleavage by endonucleases [Microparticle-based biosensors for retroviruses have demonstrated label, capture, and their combination functions. Yuan et al. developed a nucleic acid detection circuit called a ucleases . A fluorA similar biosensor demonstrated by Luo et al. capitalized on fluorescent oligonucleotide cleavage using nonspecific exonuclease III instead . Two disThe following three assays utilize ELISA-like immunoassay principles. Wang et al. proposed a relatively simple lateral flow immunochromatographic assay (LFIA) targeting the p24 protein, forming the protective shell surrounding the HIV core . The detThe number of articles is plotted against eight different virus families in The types of receptors are also shown as pie charts in Our survey results demonstrate the diverse application of microparticles towards various virus detection platforms. Nucleic acid amplification methods, immunoassays, and general hybridization reactions benefit from the high surface area-to-volume ratio of microparticles interacting with virus targets. External manipulation of magnetic microparticles presents unique strategies for mixing, separating, and immobilizing captured analytes. Finally, the convenient size of polymeric microparticles allows centrifugal separation and imaging by conventional microscopes, simplifying reaction or detection procedures. These characteristics make microparticles ideal for rapid, point-of-care diagnostic applications, which are critical for maintaining global health. Microparticle-based detection platforms still face limitations regarding their signal transduction capabilities. Within our search, electrochemical detection using microparticles remains largely uninvestigated. Certain virus families, namely Herpesviridae, Papillomaviridae, and Retroviridae, have little research into detection through microparticle-based methods. Moreover, the Filoviridae and Papillomaviridae families are highly dependent on nucleic-acid-based detection, indicating that there may be a gap in research involving antibody- and antigen-based methods for these virus families. Across all virus families, microparticles have been highly exploited for uses in analyte capture, being used in a higher or equal proportion to microparticles as labels. The ability to directly label target analytes has the potential to produce strong signals. However, labeling microparticles depends mainly on fluorometric detection. Further studies into microparticle-based virus biosensors should seek to close these discontinuities in research.


Aphis aurantii, Atrichopogon sp., Dasyhelea sp., Forcipomyia taiwana, and Trigona ventralis hoozana. Studying viruses in insects associated with economically important crops is vital to understand transmission dynamics and manage viral diseases that pose as threats for global food security. Using literature mining and public RNA next-generation sequencing data deposited in the NCBI SRA database, we identified potential vectors associated with Malvaceae plant species and characterized the microbial communities resident in these insects. Bacteria and Eukarya dominated the metagenomic analyses of all taxon groups. We also found sequences showing similarity to elements from several viral families, including Bunyavirales, Chuviridae, Iflaviridae, Narnaviridae, Orthomyxoviridae, Rhabdoviridae, Totiviridae, and Xinmoviridae. Phylogenetic analyses indicated the existence of at least 16 new viruses distributed among A. aurantii (3), Atrichopogon sp. (4), Dasyhelea sp. (3), and F. taiwana (6). No novel viruses were found for T. ventralis hoozana. For F. taiwana, the available libraries also allowed us to suggest possible vertical transmission, while for A. aurantii we followed the infection profile along the insect development. Our results highlight the importance of studying the virome of insect species associated with crop pollination, as they may play a crucial role in the transmission of viruses to economically important plants, such as those of the genus Theobroma, or they will reduce the pollination process. This information may be valuable in developing strategies to mitigate the spread of viruses and protect the global industry.This study reports the virome investigation of pollinator species and other floral visitors associated with plants from the south of Bahia:  Pollination is a fundamental ecological process that underpins the reproduction of angiosperms and ensures the production of fruits, seeds, and ultimately, the survival of numerous plant species ,2. It isInsect pollinators, particularly flower-visiting species, have been identified as important contributors to pollination services. These insects facilitate the transfer of pollen between flowers, thereby promoting fertilization and fruit set. Additionally, they contribute to ecosystem functioning, nutrient cycling, and maintenance of soil quality . The preAphis (Toxoptera) aurantii, black ants (Dolichoderus bituberculatus), mealybugs, Anoplolepis, drosophilid flies, Trigona bees, gall midges, Ceratopogonidae mosquitoes, and others  and Apis mellifera [GCA_003254395.2] genomes as alternatives to compensate for the unavailability of reference genomes for the species under investigation. The previously selected RNA-Seq libraries were aligned to reference sequences using the Bowtie2 tool, with the exception of the microbiome metatranscriptome step [The quality of each RNA-Seq library was checked using FastQC (version 0.11.9)  and filtome step . The unaome step , rnaSPAdome step , metaSPAome step , rnavira GCA_0207445.1 andome step , and MEGome step . The assGCA_0032595.2 genoThe taxonomic assessment of the assembled transcripts > 500 nt was carried out using taxonomic classification of DIAMOND Blastx (version 2.1.1)  with theThe sequences that showed similarity with viral sequences were submitted to Blastn and Blastx analysis via NCBI Blast Online  to doublViral sequences with similarity to polymerase or polyprotein coding genes were used for phylogenetic analysis. Datasets were created consisting of public protein sequences with closely related amino acid sequences and ICTV reference sequences. Global alignment was generated for each dataset using the MAFFT online tool. The alignment was visualized and trimmed in AliView . In the The complete transcriptome assembled for each insect species was assessed with the orfipy tool (version 0.0.4)  to identMalvaceae with 14.6%, followed by Poaceae with 13.5%, and Solanaceae with 8.3%  of the results were related to other species besides bees, which we referred to here as neglected pollinators and floral visitors. Overall, we observed only a few studies regarding insects involved in this important ecological service. This is evident from the analysis of the number of publications over the years. Before 1980, there was a significant shortage of publications related to this subject: less than 10 articles. Although there has been an increase in the number of articles, the maximum number of publications does not exceed one hundred, and many studies are still necessary to answer important questions on this matter . In termith 8.3% .Apis mellifera with other neglected pollinators species  and few studies related to Dasyhelea (4.2%), Trigona (4.2%), Forcipomyia (5.2%), and Atrichopogon (1.0%)  A. Therefn (1.0%) .Aphis aurantii presented a total of 147,820,116 raw reads, distributed among six libraries with an average of 24,636,686 reads per library. The assembly step produced on average 60,271 transcripts with a maximum of 73,013 sequences assembled in the library SRR11241600 (1st instar) and a minimum of 50,559 for the sample SRR11241578 (3rd instar). Regarding Atrichopogon sp., the library presented a total of 20,602,753 raw reads. After assembling non-aligned sequences against the reference genome, 169,600 contigs were identified with an N50 of 625 and an average length of 417 nt. The library that was constructed from Dasyhelea sp. presented a total of 16,591,290 raw reads. After assembling the non-aligned sequences, 150,490 contigs were identified with an N50 of 1049 and an average length of 660 nt. The libraries referring to Forcipomyia taiwana presented a total of 82,132,891 raw reads, distributed among four libraries with an average of 20,533,223 per library. The assembly step produced on average 40,541 transcripts with a maximum of 70,737 sequences assembled in the library SRR13426333, and a minimum of 13,993 for the sample SRR13426334, both libraries with samples treated with 0.25 M NaCl. Trigona ventralis hoozana had two libraries with an average of 72,269,947 reads. After assembly, an average of 135,094 transcripts were identified, ranging from 73,463,929 (SRR5601830) to 71,075,965 (SRR5601896). The N50 varied from 2754 (SRR5601830) to 2932 (SRR5601896), with an average length of 1174 nt. The assembly information for each library can be visualized in The libraries referring to Aphis aurantii, with 247 species, followed by Bacteria , Fungi , Viruses , Unknown , and Archaea  (Pseudomonadaceae (Bacteria) was the richest family in A. aurantii, with 39 species, followed by Drosophilidae , Aphididae , and Tephritidae  . Unknown families were composed of eight species , followed by Bacteria , Fungi , Viruses , Archaea , and Unknown  , Ceratopogonidae , and Pseudomonadaceae  were the richest families (Rhabdoviridae (35 species) and Phasmaviridae (13 spp.). In Dasyhelea sp., we identified 1809 species coming from Bacteria (44.2%), 1779 from Eukarya (43.5%), 358 from Fungi (8.7%), 127 from Viruses (3.1%), 13 from Archaea (0.3%), and 7 from Unknown (0.2%)  (Adintoviridae (12 spp.), Phasmaviridae (11 sequences), Baculoviridae, and Myoviridae (5 species each) , followed by Eukarya , Fungi , Viruses , Archaea , and Unknown  (Sphingomonadaceae (Bacteria) was the dominant family, with 366 species, followed by Mycobacteriaceae and Pseudomonadaceae , and Microbacteriaceae  , followed by Bacteria , Fungi , Viruses , and Unknown   and Formicidae   was the most prevalent taxon group in ., 0.2%) A. Pseudo16 spp.) B. Dicist species C. In Atr., 0.1%) D. Drosopfamilies E,F. Amonn (0.2%) G. Pseudo22 spp.) H. Regardes each) I. Bacter., 0.1%) J. In conspecies) K. Among pp. each L). Trigos, 0.2%) M. Neissespecies) N. The twfamilies N,O. The As we identified a considerable number of viral sequences in the metagenomic analysis, we further explored the virome of plants and associated insects. Therefore, we performed regular sequence similarity searches on non-redundant NCBI databases, followed by manual inspection to remove most of the common false-positive sequences, such as retroviral and phage-related elements. Our comprehensive analysis revealed twenty-three sequences showing similarity to viral sequences, four at nucleotide (nt) level and twenty-two at protein level.Aphis aurantiiAphis aurantii, we found two transcripts that share sequence similarity with unclassified elements from Riboviria. Aa_contig1 is a sequence that is 11,212 nt long, containing six ORFs. The first ORF is the longest, spanning 8247 nt, which is surrounded on the left by the 5′ UTR of 24 nt. The second, third, and fourth ORFs are 1284, 465, and 459 nt long, respectively. The fifth ORF overlaps with the fourth ORF and is 366 nt long, which is followed by the 3′ UTR of 83 nt. The Aa_contig1 showed similarity only at amino acid level, to the replicase protein sequence of Aphis glycines nege-like virus 1. The Aa_contig2 is another sequence that is 10,185 nt long encoding to four ORFs. The first ORF is 7338 nt long, preceded by a 5′ UTR of 53 nt. The second ORF of 1314 nt overlaps with the first ORF in 76 nt. The third ORF of 516 nt starts 141 nt after the end of the second ORF and is followed by the fourth ORF that contains 681 nt long succeeded by the 3′ UTR of 179 nt. This transcript also shows similarity to viruses derived from Aphis species at amino acid level, Aphis glycines virus 3 isolate 1. Aa_contig1 and Aa_contig2 contain the Alphavirus-like_MT_dom domain (IPR002588), as well as the (+)RNA_virus_helicase_core_dom (IPR027351) superimposed on the homologous superfamily P-loop_NTPase (IPR027417) in both, and Macro_dom-like (IPR043472) in Aa_contig2. Additionally, both transcripts have SP24 (IPR032441) family and the RNA-dir_pol_PSvirus (IPR007094) and Tymovirus_RNA-dep_RNA_pol (IPR001788) domains, which overlap with the DNA/RNA_pol_sf homologous superfamily (IPR043502). Aa_contig1 also has the DiSB-ORF2_chro (IPR032433) domain and Aa_contig2 also has the RNA_MeTrfase_FtsJ_dom domain (IPR002877) superimposed on the SAM-dependent_MTases_sf homologous superfamily , Picornavirus_capsid (IPR001676), and Dicistrovirus_capsid-polyPr_C (IPR014872) domains superimposed on the homologous superfamily Viral_coat (IPR029053). The RNA-dir_pol_PSvirus (IPR007094) and RNA-dir_pol_C (IPR001205) domains overlapped in the Rev_trsase/Diguanyl_cyclase (IPR043128) and DNA/RNA_pol_sf (IPR043502) homologous superfamilies. Additionally, we detected the Helicase_SF3_ssRNA_vir (IPR014759) and Helicase_SF3_ssDNA/RNA_vir (IPR000605) domains, along with the homologous superfamilies Peptidase_S1_PA_chymotrypsin (IPR043504) and Peptidase_S1_PA (IPR009003) (Iflavirus (Iflaviridae) and was closely related to Brevicoryne brassicae virus-UK with 100% bootstrap .In the samples of R009003) . Based oootstrap B. MoreovAtrichopogon sp.Atrichopogon sp., we discovered five transcripts that have similarities to elements of the order Bunyavirales. Each transcript, labeled as A_contig4, A_contig5, A_contig6, A_contig7, and A_contig8, varies in length and contains an ORF that codes for a protein. A_contig4 is 1557 nt long containing an ORF of 1536 nt with 5′ UTR of 20 nt and an open read frame at the 3′ end. According to DIAMOND Blastx, the A_contig4 sequence shares similarity with the RNA-dependent RNA polymerase (RdRp) sequence of Shuangao Insect Virus 3. A_contig5 has a partial sequence with an ORF of 2268 nucleotides with open read framings at 3′ and 5′ extremities. The best DIAMOND Blastx hit for A_contig5 is the glycoprotein sequence represented by the Wuhan mosquito virus 1. A_contig6 is 9236 nt long, and has an ORF of 8958 nt. It has a 5′ UTR of 118 nt and a 3′ UTR of 161 nt. The best DIAMOND Blastx hit for A_contig6 is the RdRp sequence of the Hubei diptera virus 3. A_contig7 is 4453 nt long, and it has an ORF of 3915 nt, with a 5′ UTR of 430 nt and a 3′ UTR of 109 nt. The best DIAMOND Blastx hit for A_contig7 is the putative glycoprotein of the Hubei diptera virus 5. A_contig8 has 1405 nt with an ORF of 1047 nt surrounded by a 5′ UTR of 117 nt and a 3′ UTR of 242 nt. It shares similarities with the nucleocapsid sequence of the Watermelon crinkle leaf-associated virus 1. Regarding the presence of conserved domains, A_contig4 and A_contig6 sequences showed RNA_pol_bunyavir (IPR007322) and RNA-dir_pol_NSvirus (IPR007099) domains  and L_PA-C-like (IPR022531). A_contig7 contained Phlebovirus_G2_fusion (IPR009878) and Phlebo_G2_C (IPR043603) domains  domain. The Capsid_Phlebovir/Tenuivir (IPR009522) family was observed in A_contig8  domain  domain .Bunyavirales. Specifically, A_contig4 was closely related to unclassified Peribunyaviridae with 100% bootstrap, while A_contig6 was closely related to Beidivirus (Phenuiviridae), with 100% bootstrap. It is important to note that elements from the order Bunyaviruses typically have segmented genomes. Based on the analyzed transcripts, it appears that A_contig4 and A_contig5 are probably the RdRP and glycoprotein segments of the same putative virus in unclassified Peribunyaviridae, respectively. On the other hand, A_contig6, A_contig7, and A_contig8 are probably the segments of the same putative virus represented by the Beidivirus (Phenuiviridae), with A_contig6 as the RdRp segment, A_contig7 as the glycoprotein segment, and A_contig8 as the nucleocapsid segment  of the Mos8Chu0 chuvirus, according to DIAMOND Blastx hit. However, there were no conserved domains for either of these contigs (Atrato Chu-like virus 1 RdRp sequence presented the conserved domains Mononeg_mRNAcap (IPR026890) and Mononeg_RNA_pol_cat (IPR014023), although A_contig9 did not present any conserved domains. Phylogenetic analysis revealed that A_contig9 was closely related to elements of the genus Doliuvirus (Chuviridae) (We found two transcripts in the library that resemble elements of the family  contigs . Only thviridae) B. This sDasyhelea sp.Dasyhelea sp. library showed similarity to the RdRp sequence of Atrato virus, which belongs to the family Totiviridae. The NCBI ORFfinder tool revealed two large ORFs containing 3063 nt and 2981 nt surrounded by the 5′ and 3′ UTRs of 536 nt and 80 nt, respectively. For one of the ORFs from D_contig11, conserved domains were detected, including the RNA-dir_pol_luteovirus family (IPR001795) and the homologous superfamily DNA/RNA_pol_sf (IPR043502) .The transcript D_contig11 of 6797 nt identified in the R043502) . Phylogeootstrap A. The coDasyhelea sp. library two transcripts that shared similarities with elements of the family Xinmoviridae. The first transcript, D_contig12, was 3812 nt long and had an ORF of 3360 nt, with a 5′ untranslated region (UTR) of 451 nt and an open read framing at 3′ extremity. The second transcript, D_contig13, was 4803 nt long, presented 4 ORFs, the largest being 1995 nt long and surrounded by a 5′ UTR of less than 684 nt. The following ORFs have 423, 399 and 468 nt, respectively, with a 3′ UTR of 41 nt. Both transcripts were similar to the RNA-dependent RNA polymerase (RdRp) sequence of Hubei rhabdo-like virus 7, according to the DIAMOND Blastx best hit. We found that both transcripts shared the Mononeg_RNA_pol_cat domain (IPR014023), although they do not show similarity at nucleotide level between themselves , Mononeg_mRNAcap (IPR026890), and Mononega_L_MeTrphase (IPR025786). Phylogenetic analysis revealed that both transcripts were closely related to elements of the genus Draselvirus (Xinmoviridae)  B. We namForcipomyia taiwanaForcipomyia taiwana libraries, we found four transcripts that were similar to elements of the family Xinmoviridae. Ft_contig14 is a sequence that is 12,795 nt long containing three ORFs. The first ORF spanning 1665 nt is surrounded on the left by the 5′ UTR of 218 nt. The second ORF is 2526 nt long, and the third is the longest ORF, presenting 6402 nt, which is followed by the 3′ UTR of 541 nt. The Ft_contig14 showed similarity only at amino acid level to the RdRp sequence of the Aedes albopictus anphevirus. Ft_contig15 was 6687 nt long, with an ORF of 6270 nt. It has a 5′ UTR of 409 nt and a 3′ UTR of 209 nt, similar to the RdRp sequence of Tolviot virus. Ft_contig16 and Ft_contig17 were similar to the RdRp sequence of the Hubei rhabdo-like virus 7 probably representing fragments of the same virus. Ft_contig16 was 1038 nt long, with an ORF of 990 nt. It had a 5′ UTR of 49 nt and open reading frame at 3′ UTR. Ft_contig17 was 855 nt long, with an ORF of 705 nt. It had a 5′ UTR of 151 nt and also showed an open reading frame at 3′ UTR. All of these sequences, except Ft_contig16  (Tolviot virus had Mononeg_RNA_pol_cat (IPR014023) and Mononega_L_MeTrfase (IPR025786) , Mononega_L_MeTrfase (IPR025786), and RNA-dir_pol_paramyxovirus (IPR016269) families. Phylogenetic analysis revealed that Ft_contig14, Ft_contig15, Ft_contig16, and Ft_contig17 were closely related to elements of the genus Draselvius (Xinmoviridae) and Unclassified . This transcript is called Ft_contig18 and has a length of 6208 nt, containing four ORFs. The first ORF spanning 1407 nt is surrounded on the left by the 5′ UTR of 181 nt. The second and third ORFs are 1104 and 825 nt long, respectively. The fourth is the longest ORF and is 1983 nt long, which is followed by the 3′ UTR of 411 nt. Regarding the presence of conserved domains, only the family Rhabdo_ncap_2 (IPR004902) was detected in Ft_contig18 (Atrato Rhabdo-like virus 2 exhibited both Mononeg_mRNAcap (IPR026890) and Mononeg_RNA_pol_cat (IPR014023) domains.In the contig16 , showed R016269) . Ft_contR025786) . Ft_contR025786) . And theassified B. We namcontig18 , while tNarnaviridae and Ourmiaviridae. The first transcript, Ft_contig19, has 2981 nt with an ORF of 2934 nt with 5′ and 3′ UTRs of 34 nt and 14 nt, respectively. It displayed similarity to the RdRp sequence of the Entomophthora narnavirus C. The second transcript, Ft_contig20, is 2285 nt long with an ORF of 2226 nt surrounded by 5′ and 3′ UTRs of 27 nt and 33 nt, respectively. It has similarity to the RdRp sequence of the Hubei narna-like virus 18. Both transcripts showed the DNA/RNA_pol_sf (IPR043502) homologous superfamily  domain, RNA_pol_mitovir (IPR008686) family, and DNA/RNA_pol_sf (IPR043502) homologous superfamily, while the Entomophthora narnavirus C did not show any domains. Based on the phylogeny of the putative viruses (Narnavirus and Ourmiavirus and were named Narnavirus forcipomyiae (NVf) and Ourmiavirus forcipomyiae (OVf), respectively.Two additional transcripts were identified in the libraries, which were similar in sequence to elements of the families erfamily . The Hub viruses A, these Orthomyxoviridae. Ft_contig21 has a length of 2478 nt and an ORF of 2433 nt surrounded by a 5′ and 3′ UTR of 31 nt and 15 nt, respectively. It is similar to the PB1 sequence of the Hainan orthomyxo-like virus 2. Ft_contig22 has a length of 2253 nt and an ORF of 2202 nt surrounded by a 5′ of 52 nt and an open read frame at the 3′ end. It is similar to the polymerase PA sequence of the Hainan orthomyxo-like virus 2. Ft_contig21 and the Hainan orthomyxo-like virus 2 both shared the RNA-dir_pol_NSvirus (IPR007099) domain and the RNA_pol_PB1_influenza (IPR001407) family, while Ft_contig22 shared the PA/PA-X_sf (IPR038372) homologous superfamily , 2nd instar nymph (SRR11241581), 3rd instar nymph (SRR11241578), 4th instar nymph (SRR11241596), adult without wings (SRR11241592), and adult with wings (SRR11241587). Un-ap2 and Un-ap1 transcripts had the highest abundance in most stages, except for 1st instar nymph. Un-ap1 had values ranging from 0.003 in the adult without wings to 17 in the adult with wings, while Un-ap1 had TPM values ranging from 0 in the adult without wings and 4th instar nymph to 54 in the adult with wings. IVa was only present in the 1st instar (TPM = 25) and in the adult without wings (TPM = 3.5). In summary, the transcriptional activity of Un-ap1 and Un-ap2 was highest in the adult with wings and lowest in the adult without wings, while IVa had the highest values in 1st instar nymph and in the wingless adult , with TPM = 2.1  and with 0.25 M NaCl (SRR13426333 and SRR13426334). DVf-PGp (TPM = 12.5) had the highest abundance in freshwater treatments. In the 0.25 M NaCl treatment, which was used to wash away microorganisms from the eggshell, all the viruses showed lower transcriptional activity  were closely related to Aphid viruses of the unclassified Riboviria (Aphis glycines nege-like virus 1 and Aphis glycines virus 3 isolate 1), while the other (Iva) were clustered with a member of the genus Iflavirus (Brevicoryne brassicae virus-UK). These viruses are known as aphid pathogenic RNA viruses, a group of aphid-infecting viruses already related to dramatic negative impacts on the growth, fecundity, development, dispersal, and functions of aphid species, such as the Aphid lethal paralysis virus (ALPV), the Myzus persicae densovirus (MpDV), and the Rhopalosiphum padi virus (RhPV) [A. aurantii, particularly during the 3rd instar nymph stage. To counteract any negative effects caused by high transcriptional activity, the virus may even be affecting the vector’s physiology, which could impact the likelihood of transmission, as observed in the Rice stripe virus (RSV) infecting Laodelphax striatellus [T. cacao plantations, since our search of the literature returned that most of these floral visitors were described as associated only with cocoa plants. Additionally, it is essential to gain a comprehensive understanding of how virus replication can interfere with the replication of other viruses and how these interactions can impact the host. It was observed that the transcriptional activity of Un-ap1 and Un-ap2 is seemingly influenced by the low transcriptional activity of IVa, and vice versa, as evident in the comparison between adults with and without wings.s (RhPV) . Despiteiatellus . HoweverAtrichopogon sp. was phylogenetically related to the family Phenuivirade, which consists of enveloped, spherical, 2–8 segmented negative-stranded RNA ((-)ssRNA) viruses with diameters ranging from 80 to 120 nm that can infect three kingdoms of host organisms  [Hubei diptera virus 3. Phenuivirade is known for comprehending various virus genera related to plant diseases, such as Tenuivirus and Coguvirus, and have different viral components associated with insect transmission and plant infection [Tenuivirus, can interfere with vital plant function, such as host cell replication, translation machinery, and ion homeostasis. For instance, the Rice grassy stunt virus (RGSV) protein p5, found in RNA segment 5, can disrupt the CBL-CIPK Ca2+ signaling network, which plays a role in regulating ion homeostasis in the plant. This can lead to a decrease in potassium content, resulting in a potassium deficit. Additionally, the protein p3 of the same virus can cause the production of an E3 ubiquitin ligase that leads to the breakdown of one subunit of plant-specific RNA polymerase IV, which is essential for RNA-directed DNA methylation [Phenuiviridae viruses have already been shown being transmitted through horizontal and vertical transmission in insects and they can be transmitted to plants through salivary glands [Atrichopogon sp. This virus is closely related to the family Chuviridae, which is made up of segmented negative-stranded RNA viruses infecting a wide variety of invertebrate hosts [Beidivirus atrichopogon found in d fungi) . It is lnfection ,60,61. Ahylation ,58. Pheny glands ,60,61. Tte hosts . Forcipomyia taiwana and is related to the family Orthomyxoviridae. This family also consists of (-)ssRNA viruses, but with 6–8 segments and three different RNA polymerases  [Orthomyxoviridae viruses also mainly infect mammals and are linked to various diseases, including influenza  [Another virus, Orthomyxo-like virus forcipomyiae, was found in n 2-PB2) . Orthomyzavirus)  and tickNarnaviridae and Ourmiaviridae were also identified in F. taiwana. Narnaviridae is characterized by a single molecule of non-encapsidated (+)ssRNA virus of 2.3–2.9 kb that encodes a single protein of 80–104 kDa with amino acid sequence motifs that are typical of an RNA dependent RNA polymerase (RdRp) [Narnaviridae, such as the Hubei narna-like virus 18, which includes information related to the homologous superfamily DNA/RNA_pol_sf. Narnaviruses have been found in plant-pathogenic fungi, as in the study by Lin et al. (2020) [Magnaporthe oryzae narnavirus virus 1 (MoNV1) in the rice blast fungus Magnaporthe oryzae [Two viruses related to pathogenic families e (RdRp) . The stu. (2020)  who detee oryzae . Additioe oryzae  has showOurmiaviridae is known for its unique fine structure, which consists of a series of discrete lengths ranging from 30 to 62 nm. It is noteworthy that the RdRp of this family has the closest similarity to the genus Narnavirus (Narnaviridae) [Ourmiaviridae infecting the cytoplasm of filamentous fungi, as noted by Fonseca and collaborators (2021) [Ourmiaviridae, is responsible for causing mosaic disease in melons (Cucumis melo) in Iran. In their study, they discussed the phylogenetic relationships between OuMV and two other RNA viruses, as well as the implications for the evolutionary significance and taxonomic position of the genus Ourmiavirus. Although there are not many studies citing Entomophthora narnavirus C, it was detected in association with insect-associated viral contigs from RNA sequencing data from samples of Thrips tabaci by Chiapello et al. (2021) [The family viridae) . The oriviridae) . Some sts (2021) . Researcs (2021)  revealed. (2021) .Dasyhelea sp. presented three new viruses, with the first one (TlVd) belonging to unclassified Totiviridae and the other two (DVd1 and DVd2) being phylogenetically closely related to the genus Draselvirus (Xinmoviridae). Totiviridae virions are non-enveloped icosahedral virions measuring approximately 40 nanometers in diameter, with a monopartite double-stranded RNA (dsRNA) genome ranging in size from 4.6 to 7.0 kilobase pairs. The genomes are typically organized into two ORFs, with ORF1 encoding a 70–100 kilodaltons capsid protein and ORF2 encoding an RNA-dependent RNA polymerase (RdRp). The replication takes place in the cytoplasm [Atrato virus, which is not typically associated with insect or plant diseases. The other two viruses were related to the family Xinmoviridae, which has single genomes segment of ~12 kb in length that encodes six/seven open reading frames (ORFs) [Aedes albopictus anphevirus , which has demonstrated the being targeted by the host interference (RNAi) response with persistent virus replication [Xinmoviridae viruses that have already been registered for mosquitoes, despite the lack of information regarding their pathogenicity.s (ORFs) . Aedes alication , is one A. aurantii was the Citrus Tristeza Virus [In our study, the data provided suggest the presence of diverse insect and plant-infecting viruses that could potentially compromise the health and survival of pollinators and floral visitors, as well as other important commercial plant species. There has been no specific research on the effect of ceratopogonids on native plants pollination or their degree of insect–plant specificity, but it is possible to infer that a reduction in their diversity could alter pollination rates in some plant species . It is aza Virus . Therefoza Virus .


The mKRAS LRR was significantly higher than wtKRAS LRR (p = 0.0145). R1 resection resulted in significantly higher LRRs than R0 resection for both wtKRAS and mKRAS , and while no significant difference was observed in the Ra and Rb LRR with wtKRAS, the Rb LRR with mKRAS (33.3%) was significantly higher than Ra LRR (5.9%) (p = 0.0289). Thus, R0 resection is sufficient for CRLM with wtKRAS; however, CRLM with mKRAS requires resection with a margin of at least 5 mm to prevent local recurrence.Local recurrence after colorectal liver metastasis (CRLM) resection severely affects survival; however, the required surgical margin width remains controversial. This study investigated the impact of KRAS status on surgical margin width and local recurrence rate (LRR) post-CRLM resection. Overall, 146 resected CRLMs with KRAS status (wild-type KRAS (wtKRAS): 98, KRAS mutant (mKRAS): 48) were included. The LRR for each group, R1 (margin positive) and R0 (margin negative), was analyzed by KRAS status. R0 was further stratified into Ra (margin ≥ 5 mm) and Rb (margin < 5 mm). Patients with local recurrence had significantly worse 5-year overall survival than those without local recurrence ( Globally, colorectal cancer is the third leading cause of cancer death, and the most common site of colorectal cancer metastasis is the liver ,2. RecenThe margin status of CRLM resection and local recurrence are closely related, and insufficient surgical margins result in local recurrence, which is a strong prognostic factor ,6,7,8. OMoreover, gene diagnosis has recently gained attention, and KRAS status is important in colorectal cancer treatment as a surrogate marker of tumor malignancy. Mutations in the RAS family genes correlate with the efficacy of anti-epidermal growth factor receptor antibodies . KRAS muTherefore, in this study, we investigated the impact of KRAS status on surgical margin width for local recurrence rates (LRR) after CRLM resection.All patients provided informed consent before inclusion in the study. This study was conducted in accordance with the Declaration of Helsinki and approved by our institutional review board .This was a single-center, retrospective study. The inclusion criteria were as follows: CRLM with KRAS status, complete macroscopic resection, description of surgical margins, and the absence of other cancers. The exclusion criteria were as follows: R2 resections, unresectable extrahepatic metastases, and BRAF mutations. BRAF mutations, while having a severe prognostic impact, are extremely rare and different in nature from RAS mutations. Therefore, they were excluded from this study. According to our previous study, CRLMs in contact with the Glissonean pedicle were also excluded because of their high LRR . Based oFirst, the surgical margin width was classified into two categories: R1 (margin positive) and R0 (margin negative). Moreover, R0 was grouped as Ra  or Rb . The cutoff value of the surgical margin width was determined using receiver operating characteristic (ROC) curve analysis. The primary endpoint was the LRR in each group, and LRRs were compared for each surgical margin width .Prior to 2015, we measured codons 12 and 13. Codons 12, 13, 59, 61, 117, and 146 were measured using the MEBGEN RASKET™-B Kit  in accordance with our revised national guidelines, from 2015 to present. In this study, all measured KRAS mutations were considered positive ,20,21.Preoperative contrast-enhanced computed tomography (CT), magnetic resonance imaging (MRI), and intraoperative ultrasonography were routinely performed. The Indocyanine green plasma clearance rate and 99 m Tc-galactosyl human serum albumin single-photon emission CT were used to evaluate the preoperative remnant liver reserve in major hepatectomy cases, as described in our previous study .We used a Cavitron ultrasonic surgical aspirator (CUSA)  for liver parenchymal dissection, and a VIO soft-coagulation system  for hemostasis. Our standard operative procedure was parenchymal-sparing hepatectomy for R0 as described in our previous study . AnatomiThe surgical margin width was determined to be the smallest distance between the tumor and the resection surface. The remaining macroscopic tumor was defined as R2 resection. Exposure of the tumor on the resection surface or microscopic infiltration within the margin was defined as R1 resection. The absence of microscopic infiltration within the surface was defined as R0 resection.Patients were followed-up every three months, and CT was performed every 6 months to investigate recurrence until 5 years after surgery. Postoperative chemotherapy was administered on a case-by-case basis. The median observation period was 1227 days (interquartile range (IQR): 639–1854 days).If recurrence was detected, it was categorized as either local recurrence, intrahepatic recurrence excluding local recurrence, extrahepatic recurrence, or peritoneal dissemination. Each resection site was precisely identified by the preoperative CT, MRI, surgical findings, and postoperative CT, which also revealed whether local recurrences were present. Local recurrence was defined as recurrence on the surface of remaining liver tissue. Intrahepatic recurrence was defined as hepatic recurrence excluding local recurrence. t-test. Kaplan–Meier plots were used to analyze the 5-year OS and log-rank tests were used to compare the groups. Statistical significance was set as p-value < 0.05.The JMP Pro13 software was used for statistical analyses . Continuous data were expressed as medians and IQRs. ROC curve analysis was used to determine the cutoff values for the univariate studies. Categorical data were analyzed using the chi-square test, and continuous data were analyzed using Student’s Local recurrence was observed in 26 tumors (17.8%). To determine the cutoff values for surgical margin width, ROC curve analysis was performed . To determine the cutoff values for surgical margin width, a receiver operating characteristic (ROC) curve analysis was performed, and 5 mm was determined as cutoff value of surgical margin.The cutoff value surgical margin was determined to be 5 mm. R0 was divided into Ra (margin ≥ 5 mm) and Rb (margin < 5 mm) after the analysis of LRR of R1 and R0. The total number of CRLMs in each group was 28, 118, 48, and 70 for R1, R0, Ra, and Rb, respectively. p = 0.0036). For mKRAS, the 5-year OS of the local recurrence group (26.6%) was also significantly worse than that of the non-local recurrence group (61.3%) (p = 0.0319).p = 0.0036). 5 year-OS of the local recurrence group was also significantly worse than that of the non-local recurrence group for mKRAS (p = 0.0319).Overall, the local recurrence group had significantly worse 5-year OS than non-local recurrence group , KRAS mutations (mKRAS), wild-type KRAS (wtKRAS).p = 0.0145). No significant differences were observed in intrahepatic recurrence, extrahepatic recurrence, and peritoneal dissemination between the two groups.The sites of recurrence of each group are presented in p = 0.0004); however, no significant difference was observed in the LRR of Ra and Rb . In wtKRAS, the LRR of R1 was significantly higher than that of R0 . The LRR of Ra and Rb in wtKRAS were 9.7% and 6.1%, respectively, which did not show statistically difference. In mKRAS, the LRR of R1 and R0 was 60.0% and 21.1%, respectively. The LRR of R1 was also significantly higher than that of R0 (p = 0.0204). Moreover, the LRR of Ra and Rb was 5.9% and 33.3%, respectively. The LRR of Rb was significantly higher than that of Ra in the mKRAS group (p = 0.0289).p = 0.0189); however, no significant difference was observed in LRR with a margin of 10 mm or more compared with a margin of 10 mm or less.We further stratified R0 resected CRLMs using 1 mm and 10 mm cutoffs and analyzed their LRRs for the Hepatectomy is the most promising treatment for CRLM. In this study, local recurrence after hepatectomy had a negative impact on OS, similar to previous studies ,6,7,8. TSeveral studies have assessed the required surgical margin width. Although a few studies have reported that the era of advances in chemotherapy and multimodal therapies may make R1 resection acceptable , R0 reseThe KRAS status is one of the surrogate markers for tumor biology. A higher risk of local recurrence and worse prognosis after mKRAS CRLM resection have been reported ,16. HoweThe LRR of mKRAS was higher than that of wtKRAS tumors, which is in accordance with previous studies reporting a high recurrence rate and poor OS in patients with mKRAS tumors. Additionally, more surgical margin width was needed for local control of patients with mKRAS tumors based on our result. Our hypothesis that mKRAS tumors associated with higher malignancy may require larger resection margin widths for local control is confirmed. Generally, mKRAS had a higher alignancy than wtKRAS tumors ,18,28. TKRAS status was an important factor in determining the surgical procedure in our study. CRLM with mKRAS requires a surgical margin of at least 5 mm but not more than 10 mm. There may be cases in which attempting to secure a 5 mm surgical margin would result in an insufficient margin and require a small anatomical resection. If the case is metachronous CRLM, examination of the previously operated primary tumor will reveal the KRAS status. Hence, if an mKRAS tumor is detected, a surgical margin width of at least 5 mm should be planned. However, for CRLM with wtKRAS, a surgical margin of 1 mm is sufficient for local control, and the surgery may be less difficult and minimally invasive. Instead of one treatment/surgical option for all, pretreatment predictors for cancer, such as liver cancer and patients’ genetic makeup, should be considered for cancer treatment for improved patient health and economy outcomes. These considerations should be made pre-surgically, with more emphasis on patients’ epi/genetic makeup ,35,36.This study had several limitations. This was a single-center, retrospective study. The sample size was small; particularly, a limited number of samples had wide surgical margin widths. Patients with tumors of varying origins were included in this study. Therefore, large prospective studies are warranted. The aim of this study was to investigate the impact of KRAS status on the surgical margin width for LRR, and the primary endpoint was the LRR. To observe local recurrence, the tumor and not the patient, should be targeted individually . If the R0 resection is sufficient for CRLM with wtKRAS; however, CRLM with mKRAS requires resection with a surgical margin width of at least 5 mm to prevent local recurrence.


New antivirals are urgently needed to treat respiratory diseases caused by RNA viruses. Unlike virus-specific antivirals, host-directed antivirals—compounds that enable cells to fight viral infections—often have the advantage of being effective against multiple viruses, including drug-resistant strains  ,2. In adCoronavirus infection can lead to significant mortality rates and economic losses in pets and farm animals . Human iHuman breast milk contains secretory IgA (sIgA), macrophages, leukocytes and other components, such as lactoferrin, lysozyme, lactadherin, linoleic acid, mucins and human milk oligosaccharides (HMOs), which contribute to the overall broad antiviral properties of human milk . There aCurrently, it is estimated that approximately one–two million people worldwide are living with HIV-2, accounting for 3–5% of the global HIV burden ,27. CompJapanese encephalitis virus (JEV), a zoonotic, mosquito-borne virus, belongs to the genus Flavivirus in the family Flaviviridae, and is closely related to West Nile virus and dengue fever virus . JEV is Mayaro virus (MAYV) is an enveloped positive-strand RNA alphavirus of the Togaviridae family that causes Mayaro fever, characterised by flu-like symptoms including fever, myalgia, arthralgia and rash . In rare


Generally, viruses can be defined as “replicators that encode structural proteins encasing their own genomes” ubiquitous in all biomes . However31, in which the aquatic and soil biomes present the greatest diversity of particles containing 7.32 x 1028 and 4.88 x 1030, respectively , direct methods allow the virus to be detected in the initial sample, through identification of the viral genome or associated antibodies . Direct methods are generally more used in virology . HTS allows the detection of known and new viral species and exploration of not only viral diversity but also the role played by these entities in host cells and biomes .Genomics can provide essential information about the stability, function, structure and evolution of viral genomes . BacteriApari and Földvári.HGT events are essential in the development of pathogenic microorganisms. HGT among different realms can occur naturally in ecosystems . In the Diadegma fenestrale, the parasitoid wasp, during oviposition, polydnavirus (PDV) are incorporated, as they reside in the ovaries as obligate symbionts. The presence of several EVEs from different PDV viral genes were observed in the host genome, being expressed even for long periods. This study demonstrates viral plasticity and the potential association of co-evolution between virus and host cells and its functional control. .Nevertheless, the description of microbiome succession and pathogen spillover is difficult to be studied, since viruses can integrate into the host genome or be integrated into the host genome as an Endogenous Viral Element (EVE) . These eKaján et al., 2020). Since viruses are subject to the same evolutionary pressures that shape the host genome composition and codon usage, they can share similar characteristics Therefore, hosts and virus coding regions tend to share common features, such as dinucleotide composition and codon usage patterns, which can be used to determine possible virus hosts . Considering the vast availability of public libraries and ongoing viral diversity studies, the number of descriptions of new viral species has increased significantly (Overall, It is well known that the relationship between viruses and their hosts is a co-evolutionary process due viruses require the host`s cellular machinery to translate their proteins. This relationship is under constant adaptation and selective pressures. The host can develop new strategies to fight viral infections, while viruses develop counter-defense measures to maintain the infection (us hosts . Anotherficantly . Neverthus hosts . Consideficantly .Using virus specific-signatures during infection to characterize host-pathogen interactions” and would like to thank all the authors who contributed to their studies. We hope that the research presented on this Research Topic will be helpful to the virus identification and characterization of host-virus interactions.We were very happy with the Research Topic “