The aim of this study is to assess the societal burden of colorectal cancer (CRC) survivorship 2–10 years post-diagnosis in terms of (1) societal costs, and (2) quality of life/utilities, and to analyze associated patient characteristics.This is a cross-sectional, bottom-up prevalence-based burden of disease study, conducted from a societal perspective in the Netherlands. In total, 155 CRC survivors were included. Utilities were measured by the EQ-5D-5L, using the Dutch tariffs. A cost questionnaire was developed to obtain cost information. Subgroup analyses were performed, based on patient characteristics and sensitivity analyses.Of all CRC survivors, 81(54%) reported no problems for mobility, 133(88%) for self-care, 98(65%) for daily activities, 59(39%) for pain/discomfort, and 112(74%) for anxiety/depression on the EQ-5D-5L. The average EQ-5D-5L utility score was 0.82 (SD = 0.2) on a scale from 0 (death) to 1 . Significant differences in utility score were found for gender, tumor stage, number of comorbidities, and lifestyle score. The average societal costs per CRC survivor per 6 months were estimated at €971 . Significant differences in costs were found for the number of comorbidities.This study shows a considerable burden of CRC survivors 2–10 years after diagnosis, in comparison with survivors sooner after diagnosis and with healthy individuals in the Netherlands.Long-term care of CRC survivors should focus on improving the societal burden by identifying modifiable factors, as summarized in the WCRF/AICR lifestyle score, including body composition, physical activity, and diet. Worldwide, the number of colorectal cancer (CRC) survivors is rising, and continuing growth is expected –3. In 20The burden of disease is often estimated on societal costs and quality of life (QoL) , 13. TheBecause the long-term survival of CRC patients has risen substantially in the last few decades, there is growing interest in this population’s quality of life . CRC surFurther, little is known about the influence of patient characteristics on the societal burden of CRC survivors . PreviouThe Dutch healthcare system consists of mandatory health insurance for Dutch citizens from private insurers, voluntary complementary insurance, and tax-funded, income-dependent long-term care by the government. Since 2006, citizens are able to freely choose private insurers for the mandatory health insurance, thereby introducing market competition . The conTo our knowledge, there is no information on Dutch societal costs of long-term CRC survivorship. Worldwide, few studies have analyzed CRC costs from a societal perspective and methodological heterogeneity and lacking transparency are present , 31. TheThis is a cross-sectional, bottom-up, prevalence-based burden of disease study from a societal perspective in the Netherlands. The study is embedded in the cross-sectional part of the “Energy for life after ColoRectal cancer” (EnCoRe) study, which assesses lifestyle and QoL of CRC survivors. The methods of the EnCoRe study have been published and are briefly described below .This study consists of patients (> 18 years of age) who have been diagnosed with and treated for stage Ι-ΙΙΙ CRC at Maastricht University Medical Center + (MUMC +) between 2002 and 2010. Patients were identified through the Netherlands Cancer Registry and recruited by mail between May 2012 and December 2013. Exclusion criteria were: (1) stage ΙV disease, (2) passed away, (3) currently no home address in the Netherlands, (4) unable to comprehend the Dutch language, and (5) presence of comorbidities that could obstruct participation. In total, 155 individuals participated in this study. The EnCoRe study was approved by the Medical Ethics Committee of MUMC + and Maastricht University, the Netherlands, and written informed consent was obtained from all participants. Participants underwent several measurements at one point during a house visit by a trained research assistant. Measurements included a general questionnaire, which was developed based on existing questionnaires by the EnCoRe research team and field experts. Additionally, the questionnaire contained questions on medical care, with recall periods of 3/6 months, depending on the estimated frequency of attendance. If estimated attendance for an activity was high, for instance visiting the general practitioner (GP), a recall period of 3 months was chosen to increase reliability. Participants wrote down all medication and supplements used in the past 6 months. The supplement packaging was checked by the research assistant.The main outcome measurements are societal costs (in 2014 Euros) and QoL/utilities. A cost questionnaire was developed by field experts, based on the steps mentioned by Thorn, and pilot tested . QoL waswww.medicijnkosten.nl and used the price per dose of the drug. If no start- and/or end-date of medication was noted, it was assumed survivors were taking the medication the full 6 months. If the frequency was missing, the lowest entered number by other survivors was assumed (0.5 unit). In case of missing data, the lowest price of the medication was assumed (e.g. lowest dose and cheapest brand). A standard price for supplements was estimated by calculating an average price per supplement from all house-brand supplements offered online by a Dutch store [Costs of individual survivors were calculated for the six months preceding the measurement and were summed (bottom-up approach). Costs were divided into healthcare sector costs, patient and family costs, and costs in other sectors. Healthcare sector costs and patient and family costs were valuated according to the most recently updated Dutch Manual for Cost Analysis in Health Care Research from 2015 . Since tch store . Medicatch store . Informach store . Travel ch store . Travel Survivors were excluded from the analyses if > 1 item on the EQ-5D-5L was missing. In case of one missing value for the EQ-5D-5L, the population mean was imputed. When medical care questions were missing, the lowest entered population number, excluding zero, was imputed. However, if the total population entered a zero, a zero was imputed. The statistical analyses were performed with SPSS version 25. Seven subgroup analyses for costs and utilities were performed, namely for: gender , age (< 70/ ≥ 70 years), tumor stage (Stage I/II/III), comorbidities (0/1/ ≥ 2), tumor subsite (colon/rectosigmoid/rectum), WCRF/AICR lifestyle score  , 44. TheEconomic evaluations are accompanied by uncertainty. In order for policy makers to correctly interpret the findings it is essential that the uncertainty of point estimates is explored . Three tData were collected from 155 colorectal cancer survivors. Four survivors were excluded, because of > 1 missing item on the EQ-5D-5L. The majority of the resulting 151 participants were male (62.3%), with a mean age of 70 years (SD = 8.7), and mean time since diagnosis of 5.7 years (SD = 1.8). The distribution of tumor stage was: 27.8% Stage Ι, 34.4% Stage ΙΙ, and 32.5% Stage ΙΙΙ. Just over half of survivors presented with 2 or more comorbid conditions (50.3%). Of all participants, 53.0% had a history of colon cancer, 4.6% had a rectosigmoid tumor, and 42.4% had rectal cancer , 1.2 for self-care (SD = 0.7), 1.6 for daily activities (SD = 0.9), 1.9 for pain/discomfort (SD = 0.9), and 1.3 for anxiety/depression (SD = 0.6). The average EQ-5D-5L utility score was 0.8 (SD = 0.2) (Table The resource use categories that showed the highest absolute number of users were medication (77.5%), travel costs 81.5%), and medical specialist 64.9%). The largest resource use per average patient was for paramedical care, with a mean of 2.9 (SD = 8.3). The estimated average societal costs per CRC survivor per 6 months were €971 . The highest costs per average survivor were observed for the categories nursing home (€204), medication (€193), and medical specialist (€141). Survivors with a time since diagnosis of ≥ 5 years showed higher total societal costs (€1007) compared to survivors with a time since diagnosis < 5 years (€888). The largest differences in costs between these two groups were for hospital and nursing home costs. Overall, the healthcare sector costs contained the largest mean costs per average patient (€849), followed by patient and family costs (€120), and then costs in other sectors (€2)  than survivors having one (€528) or zero (€316) comorbidities. There were no significant differences in costs for sex, age, tumor stage, tumor subsite, and WCRF/AICR lifestyle score  yielded a mean utility score of 0.85 , compared to the Dutch value set, which resulted in a utility score of 0.82  . RemovinThe male gender, a higher tumor stage, a lower number of comorbidities, and a higher WCRF/AICR lifestyle score were associated with higher average utility scores. The average societal costs per 6 months were €971, ranging from €0 to €32,425. Significant differences in costs were observed for having ≥ 2 comorbidities compared to one or zero.The societal costs for CRC survivors are lower compared to the average annual health expenses of the general Dutch population in 2017 (€5100), but are higher compared to the average annual health expenses of individuals with cancer in 2017 (€343) or individuals with CRC (€35) , 49. AddThe highly variable costs are in accordance with previous studies, where the majority of cancer survivors had little or no costs and a small number incurred very high costs . The meaThe QoL in this population appeared to be relatively high compared to the general population . It is wStrengths of this study are the inclusion of all societal costs, including healthcare sector costs, patient and family costs, and costs in other sectors. Also, missing data is limited. Limitations of the study are, first, measurements were performed at one point in time due to the cross-sectional design. Due to this design, no causal relationship can be established, and quality of life and costs cannot be studied over time . Second,In conclusion, this study demonstrates the considerable societal burden of CRC survivors in the Netherlands long after diagnosis. Interestingly, an association between the WCRF/AICR lifestyle score and utility score was found, implying a possible role for lifestyle factors in relation to the burden of CRC survivors. Future studies should focus on replicating these findings in a longitudinal design and study the association between lifestyle scores and cost/utilities. Additionally, future studies should further explore variables, such as particular comorbidities or medical oncologic therapy, that might explain cost/utility differences.


Encephalitis, a well-known complication of measles, is inflammation of the brain parenchyma which is mostly due to the viral invasion of neurons. It presents with a variety of symptoms ranging from mild to severe depending on the extent of the damaged neurons. The diagnosis is based on clinical symptoms such as fever, headache, altered level of consciousness, focal neurological deficits, etc. A detailed history and physical examination facilitate the diagnosis. Investigations include blood tests for measles-specific antibodies, CT, MRI, and analysis of the CSF. The management of measles-induced encephalitis mainly revolves around prevention against contracting the disease and providing supportive care if acquired. The administration of the measles vaccine is the major means of preventing this disease in childhood. Two doses are required to achieve sufficient immunity against measles, the first at the age of 12–15 months and the second at 4–6 years of age. Supportive care includes administering acetaminophen for fever, oral rehydrating salt (ORS) for diarrhea and vomiting, antibiotics for otitis media and pneumonia, and using anti-epileptics such as sodium valproate for seizures. Vitamin A can be given to prevent severe effects in children. The specific treatment would depend on the type of encephalitis the patient has developed. The measles virus, a single-stranded RNA virus, belongs to the paramyxovirus family and is well-known throughout the world for causing measles. In some individuals, it may eventually lead to the development of a variety of complications, some of which could be fatal. In recent years, measles infection has become of prime concern due to the fact that despite the wide availability of effective vaccines against the virus, a sudden upsurge in cases of measles was noted by the World Health Organization (WHO) towards the end of 2019 in states of America with a previously recognized eradication of the virus . In OctoThe measles virus, acquired via aerosol droplets, has an increased propensity for invading the dendritic and alveolar macrophages of the host’s respiratory tract . The proThe incubation period, which refers to the time from exposure to the onset of the first symptom, is approximately 11 to 12 days in measles with fever, cough, coryza, Koplik spots and measles rash being the most common presentations . Koplik Encephalitis, which is the inflammation of the brain parenchyma, is a rare but devastating complication of measles, denoting that the virus has breached the neuronal membrane or, undergoing a possible epitope matching phenomenon, the virus might have triggered an inflammatory response in the host. Measles-induced encephalitis has four distinct presentations requiring different approaches. Encephalitis may either follow an acute course  or have a chronic presentation. Chronic presentation is further subdivided into measles inclusion body encephalitis and subacute sclerosing panencephalitis (SSPE) .Primary measles encephalitis occurs during the ongoing primary measles infection and is likely to present in the first week along with the appearance of the characteristic measles rash . The proAcute encephalitis due to measles, also known as ‘post-infectious encephalitis’, is coupled with a 20% mortality rate and may manifest with fever, headaches, altered sensorium, and even later development of debilitating neurological deficit . Some stThe third type of measles inclusion body encephalitis (MIBE) also known as subacute measles encephalitis, is more likely to be observed in people with compromised immunity, and the presentation may progress from an initial altered level of consciousness to worsening seizures, eventually leading to established epilepsy and focal neurological deficits such as aphasia , hemiplegia , and ataxia (lack of coordination of movements) ,15. The Subacute sclerosing panencephalitis, a chronic demyelinating disease, is a result of persistent viral invasion of the neurons, reactivation of previously acquired measles virus, or an underdeveloped immune system as seen in children younger than 2 years of age ,16. RecePrompt diagnosis of post-infectious encephalitis is of immense importance, as it has a high tendency of giving rise to devastating sequelae. The disease can be suspected if certain clinical features are present, such as fever, headache, altered level of consciousness, focal neurological deficits, hemiplegia, aphasia, ataxia, paraplegia, or specific signs of inflammatory demyelination such as optic neuritis , which sThese clinical presentations still prove to be quite ambiguous and are insufficient to make a definitive diagnosis. Hence, a precise history can help move towards the diagnosis. Relevant questions should include the location or residence of the patient and seasonal pattern of occurrence of disease in order to rule out diseases such as Japanese encephalitis, which peaks in the rainy season. A detailed contact history should be taken, which may include any exposure of the patient to an infected individual or any travel or migration from a high-risk measles area. Enquiry related to any exposure to farm animals is also equally crucial to rule out other causes of viral encephalitis, such as flavivirus. Immunization status is sought to exclude encephalitis due to listeriosis, cryptococcus, and cytomegalovirus, as these infections are highly suspected in immunocompromised patients. Occupational history contributes to eliminating work-related causes; for example, Lyme disease or Kyasanur Forest disease .Next comes the physical examination. If conducted thoroughly, it can help to guide the evaluation of the disease in the right direction. Examination of the patient can further suggest that the infection has spread to the brain. This includes general examination of the skin, mucous membranes, lymph nodes, and central nervous system . Some spBlood tests [biochemical and hematological] to identify measles-specific antibodies, specifically immunoglobulins IgG and IgM in the serum . PreviouNasopharyngeal swabs or throat swabs and urine samples, which may also reveal presence of measles virus via polymerase chain reaction (PCR) ;Chest radiography to look for the spread of measles to the lungs in the form of pneumonia ;Electroencephalography (EEG);Computed tomography (CT);Magnetic resonance imaging (MRI) of the brain (with contrast);Single photon emission computed tomography ;Cerebrospinal fluid (CSF) analysis by lumbar puncture and cells, biochemistry, and molecular diagnostic tests (PCR)—Analysis of CSF fluid would show lymphocytic pleocytosis with normal glucose, normal or mildly raised protein, and measles virus RNA would be seen on PCR;Brain biopsy (in some cases) .The next step in the workup is to carry out laboratory testing to confirm the diagnosis, and this includes:This is a rare and fatal complication of measles infection, and a timely diagnosis is crucial. Diagnosis of SSPE is made using Dyken’s criteria which consists of two major and four minor criteria. According to the major criteria, firstly, antibodies against measles in cerebrospinal fluid are increased in levels greater than or equal to the ratio of 1:4, or antibodies found in serum are greater than or equal to 1:256; and secondly, the patient presents with typical or atypical symptoms of measles. Symptoms are called typical if the disease is rapidly growing or if the disease is long lasting and occurring on and off. Atypical symptoms consist of fits, extended stage one of disease, or an odd age of presentation that occurs in a newborn or an adult. The following are the minor criteria: (1) EEG showing periodic, generalized, bilaterally synchronous and symmetrical high-amplitude slow waves which occur regularly after every 5–15 s, which are known as periodic slow-wave complexes or “Radermecker” complexes; (2) immunoglobulin IgG levels in CSF which are higher than 20% of the sum of all proteins in CSF; (3) detection of inflammatory changes in the meninges and cerebral parenchyma necrotizing leukoencephalitis with diffuse demyelination, viral inclusion bodies in neurons, oligodendrocytes and astrocytes, neuronal loss, and astrocytosis on brain biopsy that are distinctive for SSPE; and (4) specialized molecular diagnostic test to identify the wild-type measles virus mutated genome.SSPE is diagnosed if the patient has two major and at least one minor criteria .The treatment of measles and its complications mainly revolves around preventative measures and providing supportive care if acquired. Firstly, infected individuals must be isolated, and all protective measures such as the use of personal protective equipment (PPE) such as N-95 masks, gloves and sterilized gowns by health care professionals should be implemented as per protocol. Proper handwashing technique is also encouraged.The prevention of the measles virus through vaccination is the cornerstone of its treatment. According to ACIP (Advisory Committee on Immunization Practices), it is recommended to obtain two doses of a live attenuated MMR  vaccine, with the first dose administered from 12 to 15 months of age and the second one from an age of 4 to 6 years . In areaResearch shows that the vaccine is 95% effective after its first dose and 96% effective after the second . Before The measles vaccine works by stimulating the proliferation of both antibodies and CD T4 cells, providing immunity. However, it is less effective and short-lived compared to immunity induced by the wild virus itself , as the One study also stated that if immunoglobulins are administered within 72 h of exposure to measles, there is a 75% possibility of preventing the infection. The dose to be given in the first 6 days of exposure is 0.08–0.1 cc/lb .Specific medications for measles do not exist. Only symptomatic treatment is given for the alleviation of symptoms such as fever, cough, coryza, conjunctivitis, and mouth ulcers . PatientAdministration of vitamin A is recommended in all acute cases, either as capsules or in liquid form. It is known to decrease the risk of measles-related death by 87% in children less than 2 years of age. In addition to this, it is effective in lowering the timespan of diarrhea by 2 days and of fever by 1 day in children of all age groups . In infaPatients afflicted with the measles virus can progress toward developing serious complications including diarrhea, otitis media, pneumonia, and encephalitis, which must be addressed promptly. In the United States, one or more complications occurred in around 30% of cases of measles from 1987 to 2000 , and so A study conducted in France in 2013 showed that 4 out of 32 measles infected patients developed measles-induced encephalitis, with three cases of post-infectious encephalitis and one case of measles inclusion body encephalitis . Hence, Primary measles encephalitis. Treatment majorly involves the curing of symptoms . PatientAcute post-measles encephalitis. As this form of encephalitis is immune-mediated, corticosteroids are given as first-line treatment , which aMeasles inclusion body encephalitis. Supportive treatment  is givenSubacute sclerosing panencephalitis. There is still no proven treatment for SSPE, but a combination of ribavirin (RBV), Inosiplex (Isoprinosine), and intraventricular interferon-a [IFN-a] has demonstrated to be the most effective way to deal with the disease and shorten the disease course . RibavirSince there are four types of measles-induced encephalitis and the types involve comparatively different pathologies and disease severities, the treatment plans for each type also differ. Treatment of measles-induced encephalitis depends on its type. The types and respective treatment for each are as follows:Other treatments are also available but not so widely practiced, as the data on their efficacy are yet not sufficient. Transcription or replication inhibitors of the virus RNA can transiently prevent the growth of the virus. Small interfering RNAs (siRNAs) or shRNAs are synthetic oligonucleotides which can halt the production of polymerase complex, which controls the transcription of viral RNA. The process of transcription also briefly requires heat shock proteins, such as the heat shock protein 90 (HSP90). Geldanamycin and derivatives such as 17-DMAG (17-Dimethylaminoethylamino-17-Demethoxygeldanamycin) block the activity of these proteins and might impede the transcription process . NucleosSince F protein of host cells and H protein of SSPE are involved in the fusion of the measles virus to host cells and facilitate their entry into host cells, antibodies against H protein can inhibit this fusion. There is evidence that Aprepitant, which is an antagonist of the neurokinin-1 receptor , has lowered the spread of the virus in measles-containing vaccines to the brain . As the CDC and WHO have successfully reduced the global burden of infectious diseases such as measles in countries such as the United States for the last 20 years by strategizing plans to lower its transmissibility ,53.The Measles Outbreaks Strategic Response Plan 2021–2023 was formulated by the WHO for the prevention of measles and to promote the readiness to fight measles outbreaks. It aims to conduct risk assessments every year to help formulate effective strategies against measles, to look into measles outbreaks, and to take immediate action to curb the spread of disease. It also plans to look over the main ground of the disease to fill in the immunity gap and avoid future episodes by taking appropriate measures. Surveillance of the outbreaks in 2022 can assist in identifying what is lacking in the immunization system, and suitable approaches can be thought of to bridge the immunity gaps [CDC started the “Measles and Rubella Initiative” which is intended to carry out, observe, and assess measles and rubella vaccination programs with the goal of eliminating both diseases. The plan proposes recommendations based on its operational research studies to direct activities aimed at preventing these diseases. It also aims to supply technical aids for inspecting measles and rubella breakouts, for monitoring these outbreaks, and to check vaccination status. In addition to this, it assists in initiating measles and rubella monitoring programs in different areas where these diseases have a high incidence. Furthermore, it plays the role of a reference laboratory all over the world and supplies assets on a national level. It contributes in the gathering and shipping of blood samples to test for measles using real-time polymerase chain reaction (RT-PCR) . It was Immediate recognition and diagnosis of the disease can only be possible if healthcare workers are preemptively educated with detailed information regarding the various presentations of complaints associated with measles and its complications, such as measles-induced encephalitis. Hence, being aware of the different types of measles presentations and thereby discouraging delayed diagnosis as well as initiating appropriate treatment may help in the prevention of the development of deleterious complications. Nevertheless, herd immunity remains the prime goal. Considering the highly infectious nature of the virus, attempts at executing effective and well-planned herd immunity programs is of immense importance. This can only be achieved if adequate consideration is given to prevailing diseases other than COVID-19. The realization of the fact that the measles virus is highly capable of triggering vicious responses in the brain will reduce the tendency to overlook measles vaccination campaigns, especially during the COVID-19 outbreaks. If the recent innovations and plans suggested to control measles and its related complications are carried out under strict supervision, there will be high chances of successfully eradicating the virus.


Viruses are the cause of a considerable burden to human, animal and plant health, while on the other hand playing an important role in regulating entire ecosystems. The power of new sequencing technologies combined with new tools for processing “Big Data” offers unprecedented opportunities to answer fundamental questions in virology. Virologists have an urgent need for virus-specific bioinformatics tools. These developments have led to the formation of the European Virus Bioinformatics Center, a network of experts in virology and bioinformatics who are joining forces to enable extensive exchange and collaboration between these research areas. The EVBC strives to provide talented researchers with a supportive environment free of gender bias, but the gender gap in science, especially in math-intensive fields such as computer science, persists. To bring more talented women into research and keep them there, we need to highlight role models to spark their interest, and we need to ensure that female scientists are not kept at lower levels but are given the opportunity to lead the field. Here we showcase the work of the EVBC and highlight the achievements of some outstanding women experts in virology and viral bioinformatics. Emerging and existing viruses continue to be a pressing issue in the face of an ever-growing and aging global population, posing major challenges to health science, technology and society at large. In recent years, we have witnessed the emergence of new viral diseases , the re-emergence of known diseases in new geographical areas , the increasing socio-economic burden of a wide range of lower virulence viruses , and viral diseases in animals and plants that affect security of food supply. In addition, there are a staggeringly large number of viruses in the biosphere, of which only a tiny fraction have been identified ,3. WhileThe most urgent open-ended questions in fundamental virus research require dedicated methods to be answered and can only be addressed with the help of bioinformatics. Recent biotechnological developments (such as high-throughput sequencing or metagenomics) have brought virology into the age of big data. Researchers can be overwhelmed by the sheer volume of data, and traditional methods of interpretation are prone to error. Compared to e.g., bacteriology, the required sophisticated bioinformatic tools for the analysis of virus data are only recently developed.These developments have led to the formation of a network of experts in virology and bioinformatics who are joining forces to enable extensive exchange and collaboration between these research areas. Here we present the engagement of the European Virus Bioinformatics Center and highlight the achievements of some outstanding female experts in virology and bioinformatics.http://evbc.uni-jena.de/membership-application/).The European Virus Bioinformatics Center (EVBC) was founded in 2017 and currently has 247 members see  from 140https://evbc.uni-jena.de/ (accessed on 24 May 2022)). We are publishing a monthly newsletter, informing about recent research results, upcoming events, job vacancies and further announcements, and also have a very active Twitter community spreading recent research  organized by Anne-Mieke Vandamme see .Recent biotechnological developments have led us into the age of big data and the virus bioinformatics community is constantly developing newly required sophisticated tools for extracting and interpreting relevant information from virus data. The EVBC is curating a collection of these tools. We use several properties to describe each tool: tags for the category of application , tags regarding software development , the license, tool website and respective publication, the virus families for which the tool was either designed for or validated on, the EVBC members involved. All properties are part of the search and filter function. In addition, the collection contains several presorted views to navigate through the tools, e.g., sorted by category, virus family or software type.Note that the EVBC is not maintaining the tools in the collection. However, in the future, we would like to organize assessment strategies to point researchers to the state-of-the-art tools for their analysis and point out advantages and drawbacks of the different methods.During the pandemic, bioinformatics tools designed explicitly for SARS-CoV-2 have been developed as a rapid reaction to the need for fast detection, understanding and treatment of COVID-19. The EVBC was pointing out relevant methods to combat COVID-19 and accelerate SARS-CoV-2 and coronavirus research .In addition to bringing together scientists during conferences, workshops and seminars to promote scientific exchange, the EVBC is initiating large collaborative research projects. The EU funded Marie Sklodowska-Curie Innovative Training Network VIROINF  focuses The EVBC also aims to strengthen the exchange of ideas and initiate scientific cooperation between industrial members and scientists. Both sectors have a significant interest in viral bioinformatics but often approach the subjects from different perspectives and have different requirements for the development and use of research tools. The VIROINF network is providing young scientists with training opportunities in non-academic research environments to leave them better equipped to collaborate with non-academic partners or follow non-academic research careers.Scientific results of an interdisciplinary research field are often difficult to publish in journals of either the one or the other area. Therefore, the EVBC is running a recurring special issue on virus bioinformatics since 2018. In addition, we promote special issues that are edited by our members.The first special issue on Virus Bioinformatics was published in Virus Research in 2018 covering 12 research articles and short communications. Since 2019 we are running the special issue in MDPI Viruses, covering 18 (in 2019) and 15 (in 2020) papers. The recent issue “Virus Bioinformatics 2022” is still under review and is so far covering 12 papers.The EVBC seeks to provide a supportive environment for talented researchers that is free of gender bias, for example by ensuring adequate participation of female speakers in seminars, courses and conferences. Since EVBC was founded, we have succeeded in steadily increasing the proportion of female members. However, of the 247 members of the EVBC, still only about 28% are women scientists see . The genThe more women get involved in science – or any other traditionally male-dominated field—the greater the overall knowledge in that field becomes ,15. CollWe should be aware that this is not only affecting female scientists but our society in general. What science decides to solve, for whom things are developed, and how science is conducted is strongly related to who carries out the scientific research. As an example, for years, animals used in scientific experiments were predominantly male so as not to falsify experimental results with female hormone fluctuations ,17,18. ITo keep more talented women in research, we need to ensure that they are not kept at lower levels but are given the opportunity to lead the field ,22. OfteDr. Franziska Hufsky: Coordinating EVBC activities.Franziska Hufsky studied bioinformatics at the Friedrich Schiller University Jena, Germany, where she also got her doctorate degree developing novel methods for the analysis of small molecule fragmentation mass spectra. She then joined the group of Manja Marz see  helping http://bioinfowelten.uni-jena.de/ and http://www.formeltiere.de/ (accessed on 1 July 2022)) and is in the equal opportunities team of the Friedrich Schiller University Jena.Franziska has been a founding member of the EVBC. She is compiling the monthly newsletter, monitoring the publications of EVBC members, filling the EVBC Twitter channel, maintaining the EVBC homepage including the calendar and vacancies services, and curating the collection of virus bioinformatics tools. She was in the organizing committee of all annual conferences (except 2018) and is co-editor of all special issues on virus bioinformatics in MDPI Viruses. Franziska is organizing the viruses in silico lecture series, partly organizing the ECR Viromics Webinar Series and taking care of the EVBC workshop program. She is organizing joint publications by EVBC members ,23,24, iProf. Dr. Lia van der Hoek: Anelloviruses, coronaviruses, and novel viruses.Lia van der Hoek is a virologist at the Amsterdam University Medical Center, location AMC, University of Amsterdam. She developed a novel virus discovery method, named VIDISCA, that can detect any RNA or DNA virus. Using this technique she was the first to discover human coronavirus NL63, in 2003, an endemic human coronavirus that remained unrecognized till then . It soonLia has been a founding member of the EVBC and was an invited speaker at the founding meeting of the EVBC introducing the HONOURs training network  to the cDr. Katy Brown: Virus discovery, genomics and evolution.CIAlign [Katy Brown is a postdoctoral researcher and bioinformatician at the University of Cambridge, working in Andrew Firth’s group. Her research is focused on characterizing new and poorly understood viruses. Her work involves screening high throughput sequencing data to identify RNA viruses, with a current focus on viruses of bees and associated arthropods. She has also worked on a number of projects on non-canonical translation in viruses ,36. She CIAlign , a multiShe completed her PhD at the University of Nottingham in 2014, with a focus on identifying and characterizing ancient viral elements in the DNA of modern primates and rodents ,39. She CIAlign [Katy joined the EVBC in 03/2019 and regularly attends the annual conference, where she also presented her work in 2019. She has given a lecture in the viruses in silico lecture series and her software CIAlign  is part Dr. Sofia Paraskevopoulou: Discovering new viruses in silico.Sofia Paraskevopoulou is a biologist by training who, after completing her major’s studies at the University of Crete in Greece, moved on to pursue a Masters in evolutionary biology at the University of Bonn in Germany. It was during her Masters when she discovered the world of bioinformatics and received training in writing code. She obtained a Dr. rer. nat. degree for the work that she did on virus evolution and computational virus discovery at the Charité Institute of Virology in Berlin, under the supervision of Christian Drosten.Ribozyviria [She was involved in the discovery of hundreds of novel RNA virus genomes in insects ,43, manyozyviria , a new rSofia is now a postdoctoral researcher at the Robert Koch Institute in Berlin where she has joined forces in the genomic molecular surveillance of SARS-CoV-2. Providing outbreak analytics, detecting recombinant genomes, and monitoring variants and mutations of concern are parts of her daily work.She joined the EVBC in 10/2020 and presented her work at the annual conference in 2020 . She finLuca Nishimura: Discovering ancient viruses.Luca Nishimura graduated from Hokkaido University in Japan. When she was an undergraduate student, she participated in the international Genetically Engineered Machine (iGEM) contest and proceeded with experiments on synthetic biology. After graduation, she changed her research field to virology. Currently, she is a PhD student at the Graduate University for Advanced Studies (SOKENDAI) in Japan and working on ancient viral studies.Luca’s research aim is to discover the ancient viruses more comprehensively and elucidate long-term viral evolution. To investigate ancient viruses, she has utilized whole genomic sequencing (WGS) data derived from archaeological samples, such as teeth, bones, and coprolites. She has conducted bioinformatic analyses to handle a vast amount of WGS data and tried to detect ancient viruses and comprehend the ancient viromes. Moreover, she has investigated the viral evolution using those ancient viral genomes.Luca joined the EVBC in 03/2022. She gave a poster presentation at annual conference in 2022 and won the Best Poster Award . Also, hDr. Denise Kühnert: Phylodynamics of modern and ancient pathogens.A mathematician by training, Denise Kühnert has a highly interdisciplinary background. She dove into phylogenetics in 2009 and obtained her Ph.D. in Computer Science from the University of Auckland in 2014. Thereafter, she held two consecutive postdoctoral positions in integrative biology at the ETH Zürich and in infectious diseases at the University Hospital Zurich, Switzerland. In 2018, she started her independent Max Planck Research Group at the Max Planck Institute for the Science of Human History.During her Ph.D., Denise was among the first to combine phylogenetics with mathematical epidemiology in order to use virus genomes to understand viral transmission dynamics ,49,50. FDenise joined the EVBC in 10/2017 and recently presented her research in the viruses in silico lecture series. She taught virus phylodynamics at the 23rd VEME conference in Berlin, co-organized by EVBC, and is a co-author of the overview of computational tools for SARS-CoV-2 research .Prof. Dr. Ana Barroso Abecasis: Phylodynamics and antiviral drug resistance.Ana Barroso Abecasis is Assistant Professor at Institute for Hygiene and Tropical Medicine (IHMT), NOVA University of Lisbon. She trained as Medical Doctor , has a previous degree in Pharmaceutical Sciences and specialization degrees in Bioinformatics and Microbiology. She did her PhD in Medical Sciences at the KU Leuven  working in evolutionary dynamics of HIV-1.She coordinates the research line of Global Pathogen Dispersion & Mobility of Populations at the Global Health and Tropical Medicine Research Center. She is member of several European networks of phylodynamics and antiviral drug resistance, including European Virus Bioinformatics Center, EuResist and European Society for Translational Antiviral Research. Her research interests include virology, antiviral drug resistance, bioinformatics, evolution and global health.Ana has published around 81 articles in international peer-reviewed journals, with an h-index of 26 (Google Scholar) and 19 (ResearcherID). In 2012, received the L’Oreal Portugal FCT Unesco Medal of Honour for Women in Science. She teaches at several Masters and Doctoral Programmes at the IHMT and NOVA University.VIRULIGN [ArboTyping [Ana has been a founding member of the EVBC. She has organized and taught the Workshop on Virus Evolution and Molecular Epidemiology 2018 (VEME) with Anne-Mieke Vandamme see  togetherVIRULIGN  and ArboboTyping  in the vDr. Urmila Kulkarni-Kale: Virus evolution, diversification and typing.Urmila Kulkarni-Kale is a computational virologist and senior scientist at the Bioinformatics Centre, Savitribai Phule Pune University, India. With 30+ years of experience, she has made several contributions to the fields of virus bioinformatics, immunoinformatics and phyloinformatics. Urmila is elected fellow of the Maharashtra Academy of Sciences and a member of the editorial board of the Elsevier journal ‘Infection Genetics & Evolution’.Intrigued by the diversity of viruses, Urmila has been analysing a variety of virus data to answer some of the most fundamental questions on evolution, diversification, spill overs and emergence of immune escape for viruses. Urmila has successfully built a multi-disciplinary research program and successfully applied population genetics concepts in virology to explain lineage emergence and diversification of RNA viruses. Her group has developed novel algorithms for epitope predictions , alignmeUrmila joined the EVBC in 08/2020. She is co-author of several virus typing servers ,59,60 inDr. Magda Bletsa: Characterizing viral evolutionary processes.Magda Bletsa is a post-doctoral researcher at the Department of Hygiene, Epidemiology and Medical Statistics of the National and Kapodistrian University of Athens (Greece) and an external researcher at the Department of Microbiology, Immunology and Transplantation at the Rega Institute (Belgium). She holds a Bachelor’s degree on Biochemistry and Biotechnology and a Master’s degree on Molecular Biology and Genetics. In January 2021, she received her PhD from KU Leuven (Belgium) for her work on the molecular epidemiology of HIV and hepaciviruses ,65, follHer research aims at a better understanding of viral ecology and evolution. She is particularly interested in characterizing the evolutionary processes and dynamics underlying pathogen genomic diversity across multiple scales. This goes from studying within- to between-hosts evolution, reconstructing deep and recent evolutionary events up to surveying both the virus and their hosts’ genomes ,67,68. TMagda joined the EVBC in 11/2021. She is actively attending the monthly lecture series and the annual conference. At ViBioM 2022 , she chaProf. Dr. Karin Metzner: HIV genomics and viral escape.Karin Metzner is a molecular virologist and research group leader at the Department of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, Switzerland. She has a broad and long-standing experience in the research field of viral genetics and molecular biology including next-generation sequencing of complex virus populations ,71. Her V-Pipe [Karin joined the EVBC in 02/2021. She is part of the VIROINF network . The EVBV-Pipe  in the vDr. Maryam Zaheri: Clinical metagenomics and HIV vaccine research.Maryam Zaheri is a scientist and bioinformatician in the Institute of Medical Virology of University of Zurich where she is involved in computational virus research with the main focus on metagenomics analysis of clinical samples ,75, HIV Maryam obtained her B.Sc. from University of North Carolina in Charlotte with the final project on differential gene expression analysis  and MSc Since then and until 2018 when she started her current role, she worked as a researcher at ETHZ (the group of Niko Beerenwinkel) with the focus on viral next generation sequencing analysis, as well as a bioinformatician at Roche and Novartis.Maryam joined the EVBC in 08/2019. She has attended several EVBC lectures, as well as the annual conferences since 2019. She reads the newsletters to stay updated.Dr. Stefanie Deinhardt-Emmer: Elucidating pathomechanisms.As physician, Stefanie Deinhardt-Emmer entered the field of virology during her medical thesis (PMID: 27426251). After completing clinical experience, she began her specialization in microbiology, virology, and infectious diseases. She received her first fellowships in Clinician Scientist programs, giving her the opportunity to focus on experimental research and having a research fellowship in CA, USA . She wasCurrently, Stefanie is an assistant medical director, leading a group on translational microbiology/virology at Jena University Hospital, Germany, and coordinating a BMBF collaborative project on SARS-CoV-2. Particularly important to her is the flexibility to develop new projects and the opportunity to work anywhere—at home with her children, at congresses, in the lab or in the office—to achieve her research vision: the elucidation of pathomechanisms between viruses, bacteria and the host ,89,90,91Stefanie has been a founding member of the EVBC. She was an invited speaker at the annual conference in 2019 in Glasgow .Dr. Claudia Claus: Prenatal virus infections.Claudia Claus is a senior researcher at the Institute of Medical Microbiology and Virology of the Medical Faculty of Leipzig University, Germany. She studied Biological Sciences at Martin-Luther-University Halle-Wittenberg, Germany. During her enrollment at Queen’s University Belfast, UK as a non-graduating student she attended courses with a special focus on virology. Her research stay as a visiting scientist at Georgia State University, GA, USA during her Ph.D. studies at Leipzig University deepened her interest in the study of prenatal virus infections with a focus on rubella virus. During her postdoctoral research she used an induced pluripotent stem cell-derived cell culture model to study rubella virus infection-induced alterations during the very early steps of human development . She obtClaudia joined the EVBC in 08/2019. She is attending the annual conference as well as EVBC lectures.Lore Van Espen: Human gut virome in chronic liver disease.https://microb-predict.eu/), both aiming at understanding the role of the global gut microbiome in chronic liver disease by integrating host and microbial multi-omics to identify diagnostic and/or predictive biomarkers.Lore Van Espen is a PhD researcher at the KU Leuven, Belgium, where she also obtained her BSc and MSc in Biomedical Sciences in respectively 2016 and 2018. She currently works at the Laboratory of Viral Metagenomics, Rega Institute, and is supported by an FWO fellowship. Her research focuses on the human gut virome, including eukaryotic viruses and phages, in patients with chronic liver disease. The gut viromes are studied using the in-house developed NetoVIR protocol  and a coLore joined the EVBC in 04/2021 and has attended several EVBC lectures, as well as the annual conference since 2018.Prof. Dr. Li Deng: Functional role of human virome.Li Deng studied Environmental Engineering at the Tsinghua University, China (BEng), Environmental Science at the University of Nottingham, UK (MSc), and Microbiology at the University of Bristol, UK (PhD). She studied freshwater cyanophages during her PhD under the supervision of Paul Hayes. She received post-doctoral training at the University of Arizona, US (supervisor: Matthew Sullivan) where she developed interests in using omics tools to address interactions between viruses and hosts, especially in a cultivation independent and single-cell manner. At the Helmholtz Centre Munich, Germany, she was head of a DFG Emmy Noether junior group, as well as Helmholtz Young Investigator group. She was promoted to W3 Professor of “Microbial Disease of Prevention” at the Technical University of Munich in 2022.Li conducts research in the area of phage biology. In contrast to the classical approaches, the Deng lab uses single-cell technologies, multi-omics, culture-independent techniques, and machine learning to isolate novel phages, develop phage-based therapeutics, and understand the underlying mechanisms of phage-host interactions and their impact on the human host  They appTo facilitate the acceptance of phage therapy, which uses phages to target pathogenic bacteria the Deng lab has developed highly efficient phage cocktails against multiple critical multiresistant bacteria. In addition, they promote increasing collaborations between scientists and physicians toward defining a roadmap for future translational phage research in Germany ,103.Li is a founding member of the EVBC and on the board of directors since then. She gave an invited talk at the founding conference in 2017. Li is also a principal investigator in the VIROINF network .Dr. Patricia Agudelo-Romero: Host-virus interaction phatosystem.Patricia Agudelo-Romero is a computational biologist interested in the integration of anti-viral responses of the host using different omics, such as transcriptomics and metabolomics. She holds an undergraduate engineering degree from the University of Llanos (Colombia) along with a Masters and PhD degree from The National School of Public Health of Spain and the Polytechnical University of Valencia. Since, she has held postdoctoral positions in the University of Lisbon , as well as the University of Western Australia and the Telethon Kids Institute . Through her postdocs, she has demonstrated great agility transitioning from plant to medical science and established a research platform.https://agudeloromero.github.io/EVEREST/ (accessed on 5 July 2022)). Her main purpose is to identify antiviral signatures that can be used as biomarkers to work out new treatment options in children with chronic respiratory diseases and exploring its translation into standard care.Currently, at Telethon, she is leading the bioinformatics research in the Respiratory Health for Kids team, investigating host-virus interactions in chronic respiratory diseases. She has developed bioinformatics pipelines for omics integration, viral dual RNA sequencing and viral contigs assembly and characterization  and Technical University (TU) Munich in 2003 and 2005, respectively. She obtained her Ph.D. in bioinformatics from the LMU in 2009 under the supervision of Ralf Zimmer and, after a short postdoctoral position, moved to Heidelberg University for an assistant professorship in 2010. She later returned to LMU and obtained a tenured associate professorship at the LMU in 2015.During her Ph.D., Caroline already investigated virus-host interactions, initially with a focus on protein-protein interactions for SARS  and herpHSV-1 Viewer [Caroline is a founding member of the EVBC and recently presented her research in the viruses in silico lecture series. She is co-author of the 1 Viewer  in the v1 Viewer .Prof. Dr. Irmtraud M. Meyer: Detecting functional RNA structures and RNA-RNA interactions.Irmtraud M. Meyer obtained a M.Sc. in Physics from the RWTH Aachen, Germany. She then joined Richard Durbin’s Bioinformatics research group at the Sanger Institute in Cambridge, UK, as a Wellcome Trust Prize Student and External Research Scholar of Trinity College, obtaining her PhD in 2003. After three years as a post-doc at the University of Oxford and the European Bioinformatics Institute in Cambridge, UK, she became a group leader and faculty member at the Centre for High-Throughput Biology and the Department of Computer Science at the University of British Columbia in Vancouver, Canada. Since 2016, she is a Senior Group Leader at the Berlin Institute for Medical Systems Biology at the Max Delbrück Center and Full Professor at the Freie Universität in Berlin, Germany.trans RNA-RNA interactions between different transcripts.Irmtraud is keen to investigate how the expression of genes is regulated within the transcriptome in vivo, from eukaryotes to various viruses. To that end, her research group has contributed a range of unique computational methods and analyses that are capable of detecting subtle signals within transcripts which regulate their expression in vivo ,136,137.Irmtraud joined the EVBC in 10/2020, contributing an invited lecture to the annual conference in 2020 in Basel  and partProf. Dr. Manja Marz: Bioinformatical tools for sequence analysis of RNA viruses and their hosts.Manja Marz obtained a diploma in Biology and Computer Science from the Leipzig University, Germany. After her PhD in Bioinformatics, she was immediately appointed to a group leader position at the University of Marburg, Germany, and after only two years she was offered a junior professorship at Friedrich Schiller University Jena, Germany. She became the youngest female professor to receive her full professorship three years later. During her time in Jena, she has helped shape the university by founding the Aging Research Center Jena, the Michael Stifel Center Jena for data driven Science and the European Virus Bioinformatics Center in 2017.Manja tries to bridge the world of virology and bioinformatics by (i) developing tools , (ii) applying existing tools, and finally (iii) trying to work theoretically on the existence and meaning of viruses.Together with her students, Manja developed tools to predict conserved long-range RNA-RNA interactions in full viral genomes , which hManja founded the EVBC in 2017 and has been EVBC director since then. Together with Franziska Hufsky, she is adding new features to the EVBC, from the annual meeting, to a collection and testing tools, a newsletter, etc., to the creation of a viral database and the organization of workshops upon user request.Dr. Charlotte Tumescheit: RNA structure prediction in RNA viruses, Protein Structure, MSAs.Charlotte Tumescheit is a postdoctoral researcher in Martin Steinegger’s group at Seoul National University in Seoul, Korea, where she is currently part of the team developing Foldseek , a tool CIAlign [CIAlign cleans, interprets, and visualises multiple sequence alignments. She also performed bioinformatics analyses related to virus discovery, curation of multiple sequence alignment of viruses, and protein foldability [While originally trained in mathematics, Charlotte became interested in bioinformatics and its applications to viruses and developed a tool to predict RNA secondary structure in RNA viruses during her PhD studies at the University of Cambridge under the supervision of Andrew E Firth. Here, she also developed CIAlign —togetherdability ,156,157.CIAlign is part of the EVBC tool collection. She attended the annual International Virus Bioinformatics Meeting in 2020 [CIAlign poster won the best Scientific Poster Award.Charlotte joined the EVBC in 10/2019 and  in 2020 , where hProf. Dr. Anne-Mieke Vandamme: Molecular epidemiology of viruses.Anne-Mieke Vandamme was trained as biochemist, and holds a PhD in sciences. She joined the Rega Institute at the University of Leuven in 1990, where she started a unit on virus genetic testing for clinical and epidemiological purposes. She co-founded a new division, Clinical and Epidemiological Virology; and co-started the “Institute of the future”, a transdisciplinary research incubator. She is member of the Superior Health Council of the Belgian government and of the WHO ad-hoc advisory group on COVID-19.Anne-Mieke and her team perform research on the molecular epidemiology of viruses, on virus drug resistance testing in clinical context, and on bioinformatics  tools. She supported the development of a few widely used (bioinformatics) tools: the Rega HIV Drug Resistance algorithm, the Rega HIV typing and subtyping tool, RegaDB, and Genome Detective . She is In addition to her research activities, Anne-Mieke is founder and organizer of the yearly International Bioinformatics Workshop on Virus Evolution and Molecular Epidemiology , co-initiator and co-organizer of the European HIV Drug Resistance Workshop (20th in 2022), invited professor at the Universidade Nova de Lisboa, Portugal and visiting professor at the University of Belgrade, Serbia.GenomeDetective [ArboTyping [Anne-Mieke has been a founding member of the EVBC and organized the 23rd VEME together with the EVBC. She is co-author of etective  and ArboboTyping  in the tProf. Dr. Emma Thomson: Understanding new and emerging viral infections.Emma Thomson’s lab focuses on the use of next generation sequencing and functional assays to investigate viruses that present a risk to human health in the UK and Uganda. Emma’s work focuses on understanding how viruses evolve under immune pressure and with the use of antiviral treatments. Working with the Clinical Trials Facility in the Queen Elizabeth University Hospital, Emma is the principal investigator for NHS GG&C for several vaccine trials, including the phase III Oxford/AZ ChAdOx1 vaccine, the Novavax phase III trial, the Valneva phase III trial and the COV-BOOST and DOVE studies.In March 2020, her lab sequenced the first case of SARS-CoV-2 in Scotland and continued to carry out high-scale sequencing in order to monitor introductions of virus variants in the country. Working with PHS, PHE and other partners, Emma’s lab is a member of the Genotype2Phenotype (G2P) consortium, the UK Coronavirus Immunology Consortium (CIC) and ISARIC4C with the role to investigate how changes in the genome of the virus may translate to changes in the behaviour. Additionally, she has also investigated several emerging viruses that have been detected in recent years in the Scottish population, including Ebola virus and Seoul hantavirus.GLUE [Emma joined the EVBC in 02/2022. She is co-author of GLUE  in the vDr. Evangelia Georgia Kostaki: Applying molecular epidemiology to public health.Evangelia Georgia Kostaki is a post-doctoral researcher at the Department of Hygiene, Epidemiology and Medical Statistics of the National and Kapodistrian University of Athens (NKUA) Medical School in Greece. She holds a bachelor’s degree in Mathematics, a master’s degree in Biostatistics and a Ph.D. in Molecular Epidemiology of Infectious Diseases from the NKUA, Greece. Her PhD focused on characterizing the HIV-1 epidemic in Greece using molecular epidemiology methods. The results of her Ph.D. made a significant impact to the better understanding of the dispersal patterns of HIV-1 and HIV-1 resistant strains, as well as the factors associated with the local spread in Greece ,161.Evangelia is specialized in the molecular epidemiology of HIV-1, SARS-CoV-2, and Hepatitis viruses. Her research activity focuses on the fields of epidemiology, molecular epidemiology, virus evolution, bioinformatics, and applications of molecular epidemiology to public health. In the context of her research in virology, she has used a collection of bioinformatic analysis tools that helped her analyze large datasets of viral genome sequences. Most of these tools were used for subtyping/classification, alignment, sequence similarity, phylogenetic, phylogeographic and phylodynamic analysis. Specifically, she has been involved in studies related to the investigation of how transmission networks associate with social networks, the estimation of transmission dynamics, dispersal patterns, and spatiotemporal characteristics of viral infections, as well as outbreak investigation ,165,166.Evangelia joined the EVBC in 02/2018. She is regularly reading the newsletter to stay updated and using the tool collection.Dr. Neta S. Zuckerman: Discovering and monitoring viruses that threaten public health in Israel.Neta S. Zuckerman obtained her BSc, MSc and PhD degrees in computational biology at Bar-Ilan University, Israel, developing computational methods to characterize B-cell receptor evolution and immune responses in B cell-related malignancies and autoimmune diseases. As a post-doctorate at Stanford University, USA, she developed a novel method for gene expression deconvolution and applied it to study the composition, proportions and cell-specific gene expression of cell populations in the setting of breast cancer. At Genentech Inc., USA, as a lead bioinformatics scientist, she developed and applied novel methods and pipelines to research the role of the immune system in various lung-related disorders and autoimmune diseases to understand patient heterogeneity, define bio-markers for patient groups and identify new therapeutic targets.Currently, Neta heads the Bioinformatics and Genomics center at the National Virology Laboratory, Public Health Services, Israel Ministry of Health. The center specializes in whole genome sequencing of viruses from a myriad of clinical and environmental samples, and researches viruses concerning public health in Israel, including metagenomics virome discovery for diagnostics and environmental monitoring, molecular phylogenetics to characterize viral outbreaks and gene expression studies of viral infections. During COVID, the center plays a pivotal role in SARS-CoV-2 sequencing, variant surveillance and discovery in clinical and sewage specimen in Israel.Neta joined the EVBC in 08/2018. Neta regularly participates in the annual conferences, where she gave a talk on single cell molecular dynamics of West Nile virus in 2020 .Dr. Alba Pérez-Cataluña: Monitoring viruses in wastewater.Alba Pérez-Cataluña is a postdoctoral researcher at the Institute of Agrochemistry and Food Technology (IATA-CSIC) in Spain. She has a degree in Biology and a doctorate in Biomedicine.Her studies in virology have been focused on the bioinformatic analysis of the virome present in different types of samples  using massive sequencing techniques . In addiAlba joined the EVBC in 03/2019. She is attending the annual conference and was part of the organizing committee of ViBioM 2022 . She is Dr. Janina Rahlff: Investigating phage ecology in marine systems.Janina Rahlff is a marine biologist and postdoctoral researcher and works at the Linnaeus University in Kalmar, Sweden in the group of Karin Holmfeldt. Janina investigates viral-bacterial interactions in the sea-surface microlayer, which is the uppermost, 1-mm thin layer of the water surface, with recent investigations being carried out in the Baltic Sea and the central Arctic Ocean. Microorganisms and potentially viruses in the microlayer are important in controlling gas exchange and organic matter cycling at the air-sea interface ,170. SheJanina joined the EVBC in 11/2021. She enjoys being part of this vibrant and diverse community, which she recently met during the annual conference in 2022. In July 2022, she will present her research in the ECR Viromics Webinar Series. Also, her member profile was featured in the EVBC Newsletter Vol. 57 (02/2022).Dr. Laura Miozzi: Characterizing viral biodiversity.Tuber borchii and its host plant Cistus [Laura Miozzi is a plant bioinformatician at the Institute of Sustainable Plant Protection of the National Research Council of Italy (IPSP-CNR). After the bachelor’s degree in natural sciences, she obtained a PhD on the molecular basis of the interaction between the mycorrhizal fungus t Cistus .In 2004, she started working as bioinformatician in the computational biology group at the Department of Genetics, Biology and Biochemistry, University of Torino. Her activity was mainly focused on the development of data-mining approaches for gene functional annotation, based on expression data . Since 2Laura joined the EVBC in 08/2020. She is regularly reading the newsletter and actively attending the lectures and the annual conference. She often uses the tool collection as a precious reference for her work.The EVBC is bringing together the excellence of virology and bioinformatics for more than five years now. We are constantly trying to develop our services and activities to be of valuable help to the research community, to provide numerous opportunities for scientific exchange and collaboration, and to increase the international visibility of virus bioinformatics. With all our activities we are seeking to provide a supportive environment for talented researchers that is free of any bias, be it gender, ethnicity, nationality, disability, or LGBTQ. Our female members at various career stages have made outstanding scientific achievements, such as discovering novel and ancient viruses, developing vaccines and therapeutics, understanding viral transmission dynamics, and many more. The female scientists introduced here are enriching our community with their excellent research and engagement in the EVBC activities. In the future, we would like to expand our efforts beyond gender discrimination . We are http://evbc.uni-jena.de/membership-application/).If you think that the aims and services of the EVBC could also support your research, we would be delighted if you are interested in joining and enriching our community. To become a member, please fill in the application form (


Soybean mosaic virus (SMV) through phloem was studied. Two different methods of SMV inoculation were used in the study, one was direct friction of the virus on seedling leaves and the other was based on grafting scion and rootstock to create different resistance and sensitivity combinations. Veins, petioles of inoculated leaves and rootstock stems were stained with callose specific dye. Results from fluorescence microscope observation, pharmacological test, and PCR detection of SMV coat protein gene (SMV-CP) showed the role of callose in long-distance transportation of SMV through phloem during infection of soybean seedlings. When the inhibitor of callose synthesis 2-deoxy-D-glucose (2-DDG) was used, the accumulation of callose fluorescence could hardly be detected in the resistant rootstocks. These results indicate that callose deposition in phloem restricts the long-distance transport of SMV, and that the accumulation of callose in phloem is a main contributing factor for resistance to this virus in soybean.The function of callose and its deposition characteristics at phloem in the resistance to the long-distance transportation of  Soybean Mosaic Virus (SMV) is the most common and severe disease for soybean [Glycine max (L.) Merr.] production. The disease does not only reduce yield but also seed quality. As a result, defense mechanisms to this disease have been intensively investigated using approaches of both plant physiology and plant pathology. It is now known that the success of infection depends on intracellular transport of the virus through plasmodesmata (PD) (Wang PD) Wang  and the PD) Wang . Our prePD) Wang . HoweverConsiderable knowledge on phloem structure and its role in molecular transport (Bendix and Lewis .2O2) signal was associated with the regulation of callose accumulation in restricting the cell-to-cell movement of SMV through plasmodesmata  were obtained from Grain and Oil Crops Research Institute, Hebei Academy of Agriculture and Forest Science. The other one, Nannong 1138–2, was obtained from the Soybean Institute, Nanjing Agricultural University. Both SMV strains used (SC-8 and N3) were both obtained from the Soybean Institute, Nanjing Agricultural University. SMV was propagated using the susceptible genotype Nannong 1138–2, and sap from infected leaves was used as virus inoculum based on the method described previously . The samples were then treated for 15 min in the staining solution . The treated leaves were then washed again with ddH2O and examined with a fluorescence microscope at Ex/Em = 385 nm/495 nm  and boiled for 2 min, and then washed for 5 min with ddH−1 2-DDG were used to inject leaves using the method described previously , and 500 μmol LCP gene was identified as the SMV specific coat protein gene in a previously study  and reverse-transcribed into cDNA with a reverse transcription kit . Jihuang 13 leaves inoculated with SC-8 were used as a positive control, and negative controls were non-infected leaves of either nf58 or Wuxing 2. RT-PCR assays were performed based on the method described by Li et al. ..The expression of SMV CP protein product was detected using Western blotting. Rabbit polyclonal antibody prepared with recombinant SMV CP protein expressed in E. coli was used as primary antibody at a dilution ratio of 1:5000 and detected using HRP chemiluminescence. Detailed method of western blotting was done as previously described   Fig. G–L.Fig. CP was analyzed. This gene was detected in the veins and petioles of the inoculated leaves from plants of the incompatible combination, but it was not detected in either the stem . These results indicate that, when the virus enters the phloem of resistant genotypes through grafting, callose induced by virus will deposit in the phloem, thereby limiting the long-distance transmission of the virus.CP gene based on RT-PCR was used to assess whether the virus is present in scion and rootstock. SMV-CP gene was detected in the leaves of scion nf58 and stem of rootstock Wuxing 2 below the grafting site when inoculated with SMV strain N3. However, the gene was not detected in the upper leaves nor the upper section of the rootstock stem (Fig. SMV-CP gene was detected from inoculated leaves, upper and bottom sections of stem as well as the upper leaves of Wuxing 2 when 2-DDG was used to treat the nf58 scion prior to inoculation with N3 (Fig. SMV-CP gene of the virus was detected at 96 h after inoculation. It was found that the virus was transported from the leaves of scion Jihuang 13 to the stems of rootstock Jidou 7, indicating that grafting technology successfully solved the problem of virus inoculation in phleom, and providing technical support for the study of long-distance transportation of virus through phleom and the mechanism of long-distance transportation of the virus in resistant genotypes.Phloem is the principle route for viruses transportation, studying this tissue remains a challenge (Kappagantu et al. .Sonchus yellow net virus can replicate in phloem and spread through long-distance transportation. Complete virus particles or structural proteins exist in phloem, xylem parenchyma cells, and even xylem vessels and other components, but there is no direct evidence showing that complete virus particles can be spread through long-distance transportation via xylem vessels in plants (Zhou et al. CP gene was also detected from them. The results indicated that callose accumulation was hindering the long-distance transportation of virus.The obstacles facing the long-distance transportation include the plasmodesmata mediated selective symplastic transport between companion cells and bundle sheath cells (Peleg et al. In what tissues does callose play the role in blocking the long-distance transport of the virus? It is well known that callose exists at the sieve plate of phloem (Kappagantu et al. .We have demonstrated in previous studies that callose plays an important role in restricting the cell-to-cell movement of SMV through plasmodesmata (Li et al.


Turdus merula) in Luxembourg in September 2020. The strain clustered within the Africa 3.1 lineage identified in Western Europe since 2016. Our results suggest maintenance of the virus in Europe despite little reporting during 2019–2020, rather than a new introduction.We detected Usutu virus in a dead Eurasian blackbird ( Flaviviridae, share several epidemiologic traits and cocirculate in Europe. Both viruses are maintained through a transmission cycle involving bird and mosquito vectors. Migratory birds likely play a role in long-distance spread of USUV, similarly to WNV, and in the recent introduction of the virus to Europe from Africa (West Nile virus (WNV) and Usutu virus (USUV), members of the family Turdus merula) have become a hallmark of USUV circulation in Western Europe, enabling its detection through passive surveillance , which are considered dead-end hosts  detected in Belgium, Germany, France, and the Netherlands in 2016 and 2018 (Phylogenetic analyses assigned the USUV strain from Luxembourg to the Africa 3 lineage. This lineage was first identified in Germany in 2014 . More prand 2018  Figure. The transmission of both WNV and USUV is governed by a combination of factors, such as temperature, which influences both the developmental cycles of mosquitoes and virus transmissibility (Additional information about Usutu virus Africa 3 lineage, Luxembourg, 2020


Rana grylio virus (RGV), Andrias davidianus ranavirus (ADRV), Grass carp reovirus (GCRV), Paralichthys olivaceus rhabdovirus (PORV), and Scophthalmus maximus rhabdovirus (SMRV), were measured and compared in a fish cell line. The examination of virus titers after different treatment in fish cells showed that the two iridoviruses, RGV and ADRV, had a higher tolerance to all of the environmental factors, such as they only had a decay rate of 22–36% when incubated at 37 °C for 7 days. However, the rhabdovirus SMRV was sensitive to all of the factors, with a decay rate of more than 80% in most of the treatments; even a complete inactivation (100%) can be observed after drying treatment. To address the potential threat to mammals, infectivity and limitation factors of the five viruses in Baby hamster kidney fibroblast cells (BHK-21) were tested, which showed that three of the five viruses can replicate at a low temperature, but a high temperature strongly inhibited their infection and none of them could replicate at 37 °C. This study clarified the sensitivity or tolerance of several different types of aquatic animal viruses to the main environmental factors in the aquatic environment and proved that the viruses cannot replicate in mammalian cells at normal physiological temperature.Aquatic animal viruses infect and transmit in aquatic environments, causing serious harm to the aquaculture industry and a variety of wild aquatic animals. How are they affected by environmental factors and do they represent potential threat to mammalian heath or not? Here, the effects of environmental factors (ultraviolet radiation (UV), temperature, pH, and drying) and their threshold on five epidemic aquatic animal viruses infecting amphibians and bony fish, including  Iridoviridae, Rhabdoviridae, and Reoviridae. These viruses include enveloped (Iridoviridae and Rhabdoviridae) or non-enveloped (Reoviridae) capsids, and double-stranded DNA (dsDNA), double-stranded RNA (dsRNA) or single-stranded RNA (ssRNA) genomes, which could infect specific host or non-natural host, potentially threatening aquacultural and wild aquatic animals [Viruses are major aquatic microorganism players which can cause devastating or transboundary diseases. In recent years, the rapid development of aquaculture industry has made outstanding contributions to the protection of global food nutrition and safety ,2. Howev animals . So far, animals ,10.Rana grylio virus (RGV) and Andrias davidianus ranavirus (ADRV) are ranaviruses isolated from diseased pig frog R. grylio (anura amphibian) and Chinese giant salamander A. davidianus (urodele amphibian), respectively. They are enveloped large dsDNA viruses within the genus Ranavirus in the family Iridoviridae [Ranavirus represent great threats to the aquaculture industry and wild animal populations [Ctenopharyngodon idellus) with a genome composed of 11 segments of dsRNA [Reoviridae, and mainly infects yearling and fingerling grass carp, and other cyprinids. Grass carp aquaculture accounts for a large share of freshwater aquaculture in China, and, therefore, outbreaks of grass carp hemorrhagic disease have caused great damage to the aquaculture industry every year [Paralichthys olivaceus rhabdovirus (PORV) and Scophthalmus maximus rhabdovirus (SMRV) are rhabdoviruses isolated from diseased flounder Paralichthys olivaceus and turbot Scophthalmus maximus, respectively. They are enveloped viruses with a genome consisting of a non-segmented negative-sense ssRNA within the family Rhabdoviridae [Epithelioma papulosum cyprinid (EPC) cells, which is a commonly used cell line in aquaculture research [oviridae ,12,13. Dulations ,15,16. Gof dsRNA . It is aery year . Paralicoviridae ,20. The oviridae . The fivresearch ,22,23.Considering these viruses exist in aquatic environment, the survival of the viruses are affected by different environmental factors. Which are the major environmental factors affecting the stability of the viruses? Previously, discrepant virus abundance and activities were observed in various strata of the sea along with different physicochemical factors , and virIn addition, some aquatic animal viruses have been reported to possess interspecies infection ability ,21, whicIn the present study, the five aquatic animal viruses  were selected as representatives, and used to measure and compare the effect of environmental factors such as UV radiation, temperature, pH, and drying stresses on the virus titers; we also assessed their infection ability in a cultured mammalian cell line.Siniperca chuatsi skin cell line (SCSC) that was established in our lab was maintained in L-15 medium with 10% FBS at 25 °C and was used in TCID50 assay to determine virus titers [2 [EPC cells were maintained in Medium 199 supplemented with 10% fetal bovine serum  at 25 °C . When uss titers . Baby haiters [2 .RGV, ADRV, GCRV, PORV, and SMRV were previously stored at −80 °C in our laboratory ,18,19,20−1 to 10−8). Titrations were carried out in triplicate, and subsequent experiments were conducted under this condition. Infected cells were incubated for 7 days at 25 °C except PORV at 15 °C, and the CPE was recorded. TCID50 was determined according to the previous study [For virus titration, 96-well plates containing 100 μL of confluent SCSC cells were infected with 100 μL of viruses with 10-fold serial dilutions /lgiTCID50. 50iTCID indicates the initial titer of each virus, and 50fTCID indicates the final titer obtained after different treatment.To study the effects of UV on the viruses, virus suspensions of RGV, ADRV, GCRV, PORV, and SMRV were added to sterile 6-well plates, then were treated with UV  at room temperature. Initial virus suspensions were titrated and used as 0 h data. After UV exposure for different times  , each virus was collected for titration using the TCIDTo investigate the effect of incubation at different temperatures on the viruses, virus suspensions of RGV, ADRV, GCRV, PORV, and SMRV were added into 1.5 mL EP tube and placed at 4, 15, 25, and 37 °C, respectively. Then, 150 μL of each virus suspensions were taken for titration as described above after incubating for 1, 3, and 7 days. Before incubating at different temperatures, virus suspensions were titrated and used as 0 days data.To clarify the effect of various pH on the viruses, pH values of PBS were adjusted to 3, 5, 7, 9, and 11 using 12 M HCl and 6 M NaOH, and then autoclaved for decontamination prior to provide different pH environments for the viruses. The virus suspensions were mixed with 10 times the volume of PBS at different pH, respectively, and incubated at 4 °C. A total of 150 μL of each virus suspensions were taken after incubating for 0, 1, 3, and 7 days, firstly neutralized by 1 M HEPES solution  to pH 7, and then used for titration as described above.In order to probe into the effect of drying on aquatic animal virus infectivity, 150 μL of each virus suspension was placed in individual wells of a sterile 24-well plates and allowed to dry at room temperature (25 °C) and a relative humidity (RH) of 70–80%. The lids of plates were half-opened to keep air circulating with the environmental chamber. The environmental conditions were produced in a biochemical incubator. During the evaporation period, the volume of residual virus suspension was measured and recorded at the time point , and then fresh medium was added to 1500 μL and used for titration after resuspension. Once it entered the drying phase, at each time point , samples were collected by 10~15 min incubation and rinsed with 1500 μL medium, and then used for titration.50, and then incubated at 28 °C for 1 h. After incubation, the virus suspensions were removed and a fresh medium was added. The infected cells were cultured at 28 °C and collected at 12 and 24 h post-infection (hpi) for virus gene expression detection.For determination of the sensitivity of the five viruses to BHK-21 cells, the cells were placed into 24-well plates and cultured at 37 °C to obtain a monolayer. The virus suspensions were added into each well at a dose of 100 TCID46R-3Flag) that we constructed in our previous study and that expressed EGFP were used [To test the appropriate temperature for infection of the ranaviruses (RGV and ADRV) in BHK-21 cells, two recombinant viruses , and first-strand cDNA synthesis was performed with HiScript III RT SuperMix for qPCR (+gDNA wiper) . RT-qPCR was conducted on a Quantagene q225MX Real-Time PCR Detection System . Each RT-qPCR mixture contained 5 μL of SYBR Premix (2×), 0.5 μL of forward and reverse primers (for each primer), 1 μL of cDNA, and 3 μL of ultrapure water. The RT-qPCR conditions were as follows: 95 °C for 5 min; 40 cycles of 95 °C for 15 s, 60 °C for 40 s; and a melt curve analysis from 60 to 95 °C. The MCP gene of RGV and ADRV, N gene of SMRV and PORV, and VP6 gene of GCRV were detected. The β-actin gene was used as internal control. The mRNA relative expression ratios of the treated group versus that of the control group were calculated by the 2ICP18, dUTPase, and MCP) were detected in RGV and ADRV infected cells and N gene was detected in SMRV infected cells, which were performed as described previously [For RT-PCR, three genes , the reduction in titers of viruses slowed down. Finally, the titers decreasing from low to high were, respectively, ADRV, PORV, RGV, GCRV, and SMRV. TCID50 were 102.4, 102.5, 102, 100, and 100, with a decay rate of 63%, 66%, 71%, 100%, and 100%, respectively. It indicated that SMRV was the most sensitive virus to drying stress, losing titers soon after completely dried, which suggested that drying could be an effective strategy to inactivate SMRV.In the drying assays (25 °C and 70~80% RH), the total process could be divided into two stages, the evaporation phase  and the quasi-equilibrium phase , as shown in MCP gene of RGV and ADRV and N gene of SMRV was significantly increased from 12 to 24 hpi  can be detected in different time points after infection, indicating the three viruses cannot be replicated in BHK-21 cells at 37 °C.To confirm the virus infectivity in BHK-21 cells at 37 °C, gene expressions of the three viruses, RGV, ADRV, and SMRV, were detected by RT-PCR when incubated at 37 °C. As shown in As a large group of microorganisms, viruses respond to various physicochemical factors in the environment. In this study, the effects of four environmental factors on five aquatic animal viruses were compared, and the virus infectivity on mammalian cells was analyzed. Many attractive results were observed, such as two rhabdoviruses SMRV and PORV, which showed divergent resistance to the same environmental factor; GCRV presented sensitivity to pH and resistance to UV; the enveloped dsDNA virus RGV and ADRV performed high tolerance to the tested factors; and some viruses can infect BHK-21 cell but they are limited by temperature, as shown in Rhabdoviridae. In this study, it was observed that SMRV was the most sensitive virus to UV, thermal, strong acidic pH, and drying stresses, which was significantly different to PORV, especially when encountering thermal and drying stresses. As previously reported, they, respectively, belong to genus Novirhabdovirus and Vesiculovirus. There is an NV gene in the former genome, while it does not exist in the latter. The gene has been studied in other fish rhabdoviruses, which identified its important role in supporting virus replication [PORV and SMRV are both in the family lication ,36. Howe50 significantly decreased in the weak acid or alkaline environment compared to other viruses. As previously reported, the nonenveloped reoviruses have two pH sensing proteins in capsid proteins, one of which can detach from the capsid and the other of which can undergo structural changes at low and high pH, thus affecting infection efficiency [254 exposure than the dsDNA virus and ssRNA virus genomes [GCRV was sensitive to pH, even nearly the neutral pH. There were no obvious differences among GCRV and other viruses in the face of pH 3. However, its TCIDficiency ,38. GCRVficiency ,39, whic genomes .Our results about RGV and ADRV showed that they both were relatively resistant to various tested environmental factors in this study. There have been several studies involved in environmental resistance of ranaviruses in laboratories or natural conditions ,42,43. TOn the whole, the tested viruses were all more sensitive to high temperature and low pH than low temperature and high pH, which could be related to the low stability or low enzymatic activity of the viral proteins during these situations. Although the exact mechanisms need future exploration, the unfavorable environmental factors to the viruses could be considered in disease control of aquatic animals.Iridoviridae [Cross-species infection has been reported in some aquatic animal viruses, such as ranaviruses in the family oviridae . In the The infectivity of five aquatic animal viruses under four kinds of environmental factors were tested and compared in fish cell, which showed that the tolerance of these viruses to the factors might be related to various aspects, such as the enveloped dsDNA viruses RGV and ADRV are relatively stable, but the ssRNA rhabdovirus SMRV is sensitive to UV (dose 1.5J), thermal (37 °C), and drying stresses. In addition, the aquatic animal viruses cannot replicate in mammalian cells under a normal physiological temperature, indicating aquatic animal viruses cannot propagate in mammals.


Regulatory authorities place stringent guidelines on the removal of contaminants during the manufacture of biopharmaceutical products. Monoclonal antibodies, Fc-fusion proteins, and other mammalian cell-derived biotherapeutics are heterogeneous molecules that are validated based on the production process and not on molecular homogeneity. Validation of clearance of potential contamination by viruses is a major challenge during the downstream purification of these therapeutics. Virus filtration is a single-use, size-based separation process in which the contaminating virus particles are retained while the therapeutic molecules pass through the membrane pores. Virus filtration is routinely used as part of the overall virus clearance strategy. Compromised performance of virus filters due to membrane fouling, low throughput and reduced viral clearance, is of considerable industrial significance and is frequently a major challenge. This review shows how components generated during cell culture, contaminants, and product variants can affect virus filtration of mammalian cell-derived biologics. Cell culture-derived foulants include host cell proteins, proteases, and endotoxins. We also provide mitigation measures for each potential foulant. A virus filtration step is frequently included to provide a robust size-based clearance of both enveloped and non-enveloped viruses during the manufacture of mammalian cell-derived biotherapeutics, such as monoclonal antibodies (mAbs) and Fc-fusion proteins ,2. BeforVirus filtration is different from typical pressure-driven membrane filtration processes, as the filter is designed to obtain very high levels of removal of potential virus contaminants. Further, as it is impractical to validate that there is zero carryover of any trapped virus particles, reuse of the virus filter is impossible. Consequently, these are single-use devices. Virus filters are typically run in normal flow (dead end) mode, rather than tangential flow mode used for protein ultrafiltration, since normal flow is less complex and requires only a single pump.−2) before the filtrate flux has decreased to unacceptably low levels (for operation at constant transmembrane pressure). Since biopharmaceutical manufacturing operations are still essentially batch processes, the virus filter is often sized such that the entire batch can be processed in one shift.The performance of virus filters is measured in terms of product recovery, log reduction value (LRV) of the virus (defined as the logarithm to base 10 of the ratio of the virus concentration in the feed to that in the permeate), and the productivity of the filter. Productivity is typically expressed as the volume of feed that can be processed per membrane area . Removing particulate matter such as cells and cell debris occurs at the interface between upstream and downstream unit operations. These bioreactor clarification operations are sometimes referred to as midstream processes ,14.Frequently, two polishing steps are used to remove the remaining impurities and product variants/aggregates . Resin- As shown in 10–1015 retrovirus-like particles per mL [Many mammalian cell lines produce endogenous retrovirus-like particles . These ps per mL . Removals per mL . Recent s per mL .Adventitious virus contamination is a concern in the manufacture of biologics. Validation of virus clearance is shown by conducting scale-down testing . The feeVirus filtration uses porous polymeric membranes in normal flow mode ,23,24. TThese virus filters are designed to ensure that only monomeric biomolecules with a hydrodynamic diameter less than 20 nm can pass through the pores. Much research is needed to understand how a multidomain, anisotropic mAb with varied surface moieties interacts with virus filtration membranes, prefilters, and other product monomers .Recent studies focusing on virus filtration of mAbs showed that membrane performance depends on the mAb properties , buffer conditions, membrane material, and operating pressure ,31. BuffThis section describes the major classes of foulants in virus filtration. This includes irreversible and reversible product aggregates and minor product variants that differ in their charge or hydrophobicity. Product variants arise because mammalian cell-derived biotherapeutics are heterogeneous. The product is defined based on the production process and not on a single molecular species. Product variants with different post-translational modifications can have different hydrophobicity, charge, and conformations. If present, HCP, proteases, and nucleic acids can also foul the virus filter.Aggregation is a typical occurrence with mAbs and other therapeutic proteins. Several pathways have been proposed to describe the aggregation of proteins. They include agglomeration of monomers in their native states, aggregation of conformationally altered or chemically modified monomers, nucleation, and surface-induced aggregation ,35,36. SPhysical or chemical perturbations that put a strain on the native conformation of biotherapeutic proteins such as mAbs can result in clipping or aggregation . Such coInterfacial damage can also affect the stability of a product monomer, especially at the air–liquid interface, which induces nucleation and aggregation . SurfaceReversible aggregates are usually a precursor to nucleation , followeAggregation can occur through different pathways, resulting in aggregates that are reversible or irreversible ,37. mAb Rayfield et al.  investigWhen product dimers and trimers undergo further aggregation, they attain a critical mass where the aggregate can no longer remain soluble. These large aggregates then precipitate out of the solution . The preIrreversible aggregation is prevalent with denatured product monomers ,37. ChemHCPs feature significantly in the downstream processing of protein-based therapeutics . HCPs inEnzymatic HCPs (host cell proteases) can clip or denature product monomers, expose hydrophobic residues and charged moieties, and alter the product’s biophysical properties. Denatured product monomers with exposed residues induce virus filter fouling by adsorptive processes in addition to mAb–mAb and mAb–HCP association. Host cell proteases have been reported to result in the fragmentation of mAb products, with increased susceptibility to nucleation and aggregation . HoweverHCP diminishes the biotherapeutic quality of biotherapeutic products and increases downstream processing costs. If HCPs are not sufficiently removed, they could potentially induce flux decay during virus filtration. HCPs have a range of biophysical properties, such as pI (2–11) and mass (10–200 kDa), which can be used to separate the HCP from the biotherapeutic ,66. ProtEndotoxins or lipopolysaccharides (LPS) are contaminants that can enter the process through growth media or other cell culture additives used in mammalian cell cultures. LPS are produced by Gram-negative bacteria, commonly used in recombinant DNA production ,69,70. ELPS have been reported to have a high affinity for some biotherapeutic proteins . LPS andLPS have molecular masses ranging from 3–40 kDa, which vary due to their polysaccharide chain lengths ,70. EndoThe charge variant profile is a critical quality attribute of mAbs  and Fc-fThe net charge and surface charge distribution of glycoproteins change with buffer pH ,81,82. THydrophobic interaction is the preferential association of non-polar residues in aqueous media . Amino aThe glycan appendages of glycoproteins also contribute to the final stable conformation, and glycan variation can introduce minor hydrophobicity variations. Careful handling and mild changes in formulation conditions will reduce the formation of conformational variants, which could foul virus filters or induce product aggregation.Monoclonal antibodies and Fc-fusion proteins consist of amino acids in specific sequences that form secondary, tertiary, and quaternary structures. Sequence variants arise due to genetically unprogrammed amino acid substitutions, omissions, or insertions during biosynthesis . SequencThe primary structure (amino acid sequence) of a glycoprotein can determine intermolecular, monomeric association, and aggregation propensity . Even miE. coli expresses mostly insoluble, non-glycosylated variants [mAbs, antibody fragments, bispecific antibodies, and Fc-fusion proteins are expressed in mammalian cells such as Chinese Hamster Ovary (CHO) cells for pharmaceutically relevant glycosylation profiles . Flynn evariants . Hybridovariants ,92,93.During cell culture and harvesting operations, expressed glycoproteins are usually not uniformly glycosylated ,94,95. GGlycans are hydrophilic oligosaccharide moieties typically appended to glycoproteins in the cell during glycoprotein synthesis . GlycansGlycan type and abundance can alter the product’s biophysical properties. Several studies have looked at the stability of different mAb glycoforms. These results show that aggregation is more prevalent in unglycosylated mAbs since glycans modulate aggregation ,112. FurEven though the support structure of the virus filter can function as an inline prefilter, significant fouling is often observed due to the product- and process-related foulants listed above that could be present in the feed stream. Standard practice involves the inclusion of a virus prefilter to remove these contaminants. Virus prefilters may rely on one or more mechanisms of action for the removal of foulants.A prefilter, often inline, is added upstream of a virus filter to increase permeate flux and productivity. The improvement in performance depends on the biotherapeutic product properties, prefilter material, and buffer conditions . The mecThe Sartobind S and Q are commonly used as polishing steps, usually before the virus filter. They are run as a separate unit operation (not inline) and can also provide significant virus clearance by adsorption. Brown et al. indicate that virus filter throughput may be increased by adsorptive ion exchange membrane prefiltration . ConsequWickramasinghe et al. opined that trace amounts of aggregates that have a diameter less than 50 nm play a significant role in virus filtration membrane fouling . These sEndotoxins can be removed using hydrophobic prefilters, which bind the phosphorylated lipid moiety, or using anion exchange prefilters to capture the polysaccharide moiety ,72,116. Hydrophobic prefilters require a moderate/high salt content (ionic strength) to reduce the product’s solvation layer, enabling exposed hydrophobic patches to adsorb on the hydrophobic prefilter. Hydrophobic interaction prefilters can be effective in removing product variants with different hydrophobicity, as well as some of the more hydrophobic product aggregates.Ion exchange prefilters are helpful in the downstream removal of HCPs due to the pI difference between mAbs and most HCPs. DNA is strongly negatively charged in aqueous solution and can be effectively removed using anion exchange membranes during polishing operations ,116.Multimodal prefilters are useful to filter out foulants that cannot be removed by ion exchange, size exclusion, or hydrophobic interaction-based prefilters alone. These multimodal prefilters include the three prefilters from MilliporeSigma, as shown in Monoclonal antibody properties are highly dependent on the buffer conditions and excipients that are part of the formulation. Excipients are non-drug substance components of the formulation. During high-throughput screening of mAbs for optimum buffer conditions, a specific buffer type and composition may be found to inhibit aggregation and mitigate fouling of virus filters. Phosphate, acetate, and tris buffers may work for some biomolecules, while viscosity-inhibiting buffers may be preferred for highly concentrated mAbs. Arginine reduces mAb monomeric self-association, non-specific membrane interactions, and mAb aggregation . ExcipieAchieving high levels of virus clearance for mammalian cell-derived biotherapeutics will continue to be a challenge. There is a continuing need for better virus filters that maximize productivity, flux, and LRV for batch processes. As biomanufacturing moves towards continuous manufacturing platforms, there will be a need to develop new virus filters. Unlike current virus filters, which are designed to process a product batch in one shift, in continuous biomanufacturing, the virus filter will be run for much longer times and likely at much lower filtrate flux/transmembrane pressure. Further development of virus filters for continuous operations will be needed.There is a growing demand for virus particles and virus-like particle-based vectors for delivery of gene therapies and vaccines. Virus particle-based delivery systems such as attenuated, recombinant, infectious, and inactivated virus particles, as well as virus-like particles and even subunits of virus particles, are highly effective therapeutics. However, downstream purification of these new therapeutics is challenging. Future virus filter designs will need to be optimized for these emerging therapeutics.