Flaviviridae, Parvoviridae, Phenuiviridae, and Rhabdoviridae. Notably, the Dabieshan tick virus (DBTV) of the family Phenuiviridae was prevalent in these ticks, with the minimum infection rate (MIR) of 9.09%, higher than previously reported in numerous provinces in China. In addition, sequences of tick-borne viruses of the family Rhabdoviridae have first been reported from the border area of Liaoning Province, China, after being described from Hubei Province, China. This research furthered the insight into pathogens carried by ticks in the northeastern border areas of China, offering epidemiological information for possible forthcoming outbreaks of infectious diseases. Meanwhile, we provided an essential reference for assessing the risk of tick bite infection in humans and animals, as well as for exploring into the evolution of the virus and the mechanisms of species transmission.Ticks play a significant role in transmitting arboviruses, which pose a risk to human and animal health. The region of Liaoning Province, China, with abundant plant resources with multiple tick populations, has reported several tick-borne diseases. However, there remains a scarcity of research on the composition and evolution of the tick virome. In this study, we conducted the metagenomic analysis of 561 ticks in the border area of Liaoning Province in China and identified viruses related to known diseases in humans and animals, including severe fever with thrombocytopenia syndrome virus (SFTSV) and nairobi sheep disease virus (NSDV). Moreover, the groups of tick viruses were also closely related to the families of  In recent decades, changes in the global natural ecology and frequent international trade have expanded the geographical distribution of tick species, while the number of potentials or known tick-borne pathogens continues to increase . Tick-boNext-generation sequencing (NGS) and metagenomics analysis have demonstrated superiority in discovering novel viruses in ticks and various invertebrate vectors and exploring viral evolution. These viral sequences reveal evolutionary and genetic features of the genome, and some of them appear likely to be the phylogenetic ancestors of infected vertebrates. Not only does this reveal the diversity of viruses, but it also contributes to a greater extent to the understanding of the microbiome and thus to public awareness of the viral community . As the Haemaphysalis longicornis represents the dominant tick species in Liaoning Province, carrying more than 50 species of pathogens , a total of 561 ticks were collected using the drag-flag method from 14 sites in four border cities in eastern Liaoning Province, namely Dandong city , Fengcheng city , Tieling city , and Benxi city  . Ticks w® SE Viral DNA/RNA Kit according to the manufacturer’s protocol by sieving 250 μL filtrate into a 1.5 mL centrifuge tube. Total RNA was eluted in 40 μL of Rnase-free H2O, and 1 μl RNase inhibitor was immediately added to each sample. One micro liter samples were taken and quantified using a Qubit® 2.0 fluorometer . The extracted RNA was stored in a − 80°C refrigerator for later use.According to the tick collection area, 561 ticks were divided into 58 groups, respectively, into 1 mL phosphate buffered saline (PBS), adding zirconium beads, and grinding in a frozen grinder at 60 hz for 10 min. After centrifugation at 12,000 × g for 5 min, 300 μL tick supernatant was absorbed into 0.45 nm filter membrane and centrifuged at 12,000 × g for 3 min to remove excess tick cells and bacteria as much as possible. Total viral nucleic acid was extracted using the OMEGA E.Z.N.A.® rRNA to remove kits rRNA. Following the manufacturer’s instructions, set the condition as follows: 94°C 7 Min, 75°C 2 Min, 70°C 2 Min, 65°C 2 Min, 60°C 2 Min, 55°C 2 Min, 37°C 5 Min, 25°C 5 Min, and hold at 4°C. Next, the NEBNext® ultra II directional RNA library prep Kit for Illumina was used to construct the library according to the manufacturer’s instructions. First, the de-host library samples were fragmented, and then the first strand of cDNA and the second strand of cDNA were synthesized. The end of the next generation phosphorylation level, and dA short tail and the connection joint. To make the molecules attached to the adaptor available for subsequent PCR reactions, the USER® enzyme was used to remove uracil from the NEBNext adaptor ring structure. Finally, PCR enrichment was carried out using the NEBNext® multiplex oligos for Illumina (96 index primers) Kit, followed by RNA library purification.Depending on the RNA concentration, 3–4 groups of tick RNA with the equivalent mass of total RNA belonging to the same collection site were mixed to generate a total of 18 RNA library pools. Enrichment of RNA, then use NEBNext−5  and protein (NR) database using BLASTx and BLASTn with an E-value cut-off of <10−5 . The hig® qPCR SYBR Green Master mix (Low Rox Plus) kit , the cycling program was set to pre-denaturing at 95°C for 5 Min, (40 cycles) denaturing at 95°C for 10s and finally annealing at 60°C for 30s. The positive viral sequences obtained by RT-PCR were mapped to the target sequence using CLC Genomics Workbench 12.0 to determine the sequencing depth and coverage H. longicornis were analyzed by morphological and molecular biological identification, and sequences from the sequencing results correlated with all COI barcode records in the NT database and the Barcode of Life Data System (BOLD). Eighteen library pools were generated for high-throughput sequencing, yielding a total of 709 million raw reads, with 318 million clean reads retained. The viral sequences were analyzed based on BLASTx and BLASTn, and contigs were aligned with NT or NR databases with E values ≤10−5 , Fengcheng city , and Tieling city (DBTV-TL), respectively , also identified using metagenomic sequencing analysis in a patient with a history of tick bites in northwest China, caused symptoms such as headache, fever, and erythema  in eastern Africa , 5,052 (M), and 1501 (S) bp, respectively  coveringectively . The NSDectively –D. Notabectively . NSDV coFlaviviridae spread widely around the world, mainly by arthropods , and several important tick-borne viruses include Tick borne encephalitis virus (TBEV), West Nile virus (WNV), Japanese encephalitis virus (JEV). Also, in recent years, new unclassified segmented flavi-like viruses such as Jingmen tick virus, ALSV, Yanggou tick virus, and others have been discovered , the VP1a and VP1b proteins, and was specific for JMVs . A phyloRhabdoviridae family and designated it Rhabdoviridae sp. strain RhV/LN (RhV/LN). The phylogenetic analysis of RdRp revealed  and Densovirinae VP1 (DNSV-VP1). At the amino acid level, DNSV-NS1 shared 73.52% amino acid identity with QTE18654.1TKDSV1, and DNSV-VP1 shared 91.47% amino acid identity with QTE18655.1TKDSV2  compared to previous tick DBTV surveys in Shandong, Yunnan, and Guizhou provinces. DBTV most likely predominates as a novel tick-borne virus in human populations. In addition, RhV/LN was reported for the first time in Liaoning Province, China, after being reported in Hubei Province, China. This research, therefore, not only enriches the diversity of tick-borne viruses but also reaffirms the need for continuous surveillance of tick-borne viruses in the border areas of northeastern China.Based on the results of this high-throughput sequencing, a total of 17 species infecting human viruses or potentially pathogenic families were detected in Flaviviridae, Nairoviridae, Phenuiviridae, and Retroviridae had reads in each library, while some tick-related virus families, such as Adintoviridae, Hepeviridae, had reads in only a minority of pools. The differences in the number of viruses detected compared to previous investigations of H. longicornis in forested areas of Liaoning Province seem to be due, on the one hand, to differences in the time and place of tick collection and, on the other hand, to the fact that some of the low concentrations of viruses were degraded under non-anthropogenic conditions . Although no serological evidence of NSDV infection in humans and animals has been temporarily discovered in China, NSDV has been successfully isolated from various host animals  and humans in Africa and South Asia. Haemaphysalis intermedia ticks have been considered as a possible vector for its transmission , while annotating the sequencing results as contigs for NSDV, with the sequences obtained covering the entire ORF of the genome. Phylogenetic analysis evidences the clustering of the NSDV identified for this work with previously studied strains from China, but also some sequence differences while reflecting the genetic diversity of the virus across countries. It is noteworthy that in the phylogenetic analysis, Chinese NSDV strain appears to be more closely related to the African strain than to the Indian strain in evolutionary terms. The results resembled those of Yang L et al. and suggested a potential evolutionary route for this virus  and systemic diseases such as hemorrhagic fever , with no specific drugs or vaccines currently available for the diseases , some of which may be unique hosts or may act as intermediate hosts for transmission to other animals or plants. The delineation of genera also requires significant differences in genome sequence and structure, antigenicity, and ecological characteristics , as recommended by ICTV.Rhabdoviridae family.The H. longicornis is a common tick species in northeastern China with a wide geographic distribution and host diversity, transmitting at least 30 human pathogens and serving as a vector for a variety of pathogens of medical and veterinary significance (Differences in environment and tick species may contribute to the diversity and abundance of viruses carried on ticks. ificance . In thisThe recurrence of tick-borne viruses (TBDs) and the frequent occurrence of novel TBD events in recent years have made the control and prevention of TBDs difficult. On the one hand, the abundance of forests in Liaoning and its mild and humid climate provides a favorable environment for ticks to reproduce and grow, and the frequent trade exchanges with Korea and Japan in Liaoning Province provide the basis for tick species diversity. On the other hand, it is mainly due to our limited knowledge of the background of tick-associated viral diseases and the complex mechanisms of their transmission in nature. To achieve these goals, firstly, virus surveillance should be strengthened in susceptible areas and regular testing in non-susceptible areas, and secondly, tick bites in outdoor areas such as jungle areas should be prevented by reducing skin exposure. Finally, given that human health relates to many aspects of the modern living environment, including the health of plants and animals, ecosystem stability, and natural climate change, future research should include modeling of infection and transmission and must adhere to a “one health” strategy .H. longicornis in Liaoning Province, identifying the known viruses SFTSV and NSDV that cause disease in humans and animals, and our results also confirm the presence of rhabdo-, phlebo-, flavi-like, and parvovirus sequences in ticks. Not only does it validate the desirability of virus surveillance in areas where tick-borne diseases are endemic, but it also provides information for possible future outbreaks of infectious diseases. At the same time, it offers an excellent reference for assessing the risk of infection by tick bites in both humans and animals, as well as insight into the evolution of the virus concerning mechanisms of species transmission.In conclusion, this study applied mNGS to systematically examine the virulence group carried by the dominant tick species The data presented in the study are deposited in the Genbank and repository, accession number OQ716537-OQ716539, OQ716521-OQ716522, OQ716519-OQ716520, OQ716534-OQ716536, OQ716542, OQ716540, OQ716541.YB and YL designed the experiments and performed the experiments. YT and XH provided reagents and materials. YB, YL, JL, and FT analyzed the data. YB drafted the manuscript. All authors critically read and revised the manuscript.This research was supported by the National Key Research and Development Program of China .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Plant pathogenic viruses in agricultural systems significantly reduce crop yields and pose a serious threat to global food security systems. However, the advent of “viromics” in biology has uncovered a plethora of viruses whose effects on ecosystems and food security have not been significantly studied. Studies on viruses are attracting increasing attention in agriculture because of their ability to directly infect crops and the wide-ranging ecological consequences they can have on agricultural ecosystems. Viruses can help plants survive biotic and abiotic stresses under certain conditions. In addition, viruses can interact with bacteria, weeds, pathogenic and symbiotic fungi, insect pollinators, and other species in ways that benefit or harm final crop production.Viruses in agricultural systems: Interactions with plants, insect pollinators, and fungi” was developed under the sections “Virology” and “Microbe and Virus Interactions with Plants,” which belong to the journal “Frontiers in Microbiology.” The distribution, diversity, transmission, pathogenesis, and control of viruses infecting plants, fungi, and insects in agricultural ecosystems were considered for this topic. However, the final article collection was expanded to include viruses infecting other host species in agroecosystems not listed in the title, considering host-shifting of viruses and phylogenetically related cross-kingdom viruses. This topic has accepted eight original research articles and one review article covering a range of topics related to viruses in agricultural systems. Of these, five of the original research articles focused on the molecular processes of viral infections in different hosts in agricultural systems.The Research Topic “Gong et al., it was shown that protein lysine acetylation (Kac), an essential post-translational modification in eukaryotes or prokaryotes, has a major impact on the tomato spotted wilt virus (TSWV) infection of Nicotiana benthamiana. Gong et al. discovered around three thousand sites linked to acetylated lysine in approximately two thousand proteins by analyzing the acetylation status of proteins in virus-free and TSWV-infected N. benthamiana leaves. They also found that after the viral infection, 408 sites on 294 proteins were positively regulated and 284 sites on 219 proteins were negatively regulated. Altogether, 35 conserved motifs were recovered, and K*R was the most prevalent combination, accounting for 1,334 (31.63%) of the enriched motifs. According to their bioinformatics study, most proteins with lysine acetylation sites were found in the cytoplasm and chloroplast and were engaged in biological activities such as cellular and metabolic ones.In the work by Xu et al. investigated the influence of the Fragaria vesca LHC II-1L protein (FvLHC II-1L) in strawberry vein banding virus (SVBV) infection. They found that the movement protein SVBV P1 interacted with FvLHC II-1L in vitro and in vivo. SVBV-infected epidermal cells of N. benthamiana leaves displayed co-localization of SVBV P1 and FvLHC II-1L; however, F. vesca had elevated FvLHC II-1L protein synthesis. Additionally, Xu et al. discovered that FvLHC II-1L efficiently boosted SVBV P1 to compensate for the intercellular movement of movement-deficient potato virus X (PVXΔP25) and the systemic movement of movement-deficient cucumber mosaic virus (CMVΔMP). Further, it was shown that the overexpression of FvLHC II-1L can enhance SVBV infection of F. vesca via interacting with the SVBV P1 protein.Chlorophyll a/b-binding protein of the light-harvesting complex II type 1 like (LHC II-1L), which largely controls the integrity of the electron transport system, is an essential component of photosynthesis. Sun et al. challenged the notion that virions of a virus that infects the host have the same viral genome and properties by examining the genetic variability of the virions of the white spot syndrome virus (WSSV), a DNA virus that affects crustaceans. They found that the WSSV population was made up of a 1:3 ratio of Type A (containing the WSSV lncRNA-24) and Type B (carrying the wsv195 gene), where there are two nucleotide changes between these two types. All virus-infected cells and tissues from different hosts showed a consistent 1:3 structure in viral populations. The study showed that type A WSSV virus infection was boosted by WSSV lncRNA-24 by interacting with WSSV miRNAs in shrimp, but type B WSSV required the wsv195 gene for viral infection. When Type A or Type B WSSV was eliminated from the WSSV population, the copy number of the virus decreased by 100 times in shrimp, and infection was prevented by the simultaneous elimination of both WSSV types. The complementary effects on the WSSV population and the accomplishment of the viral infection by two distinct WSSV types expressing various operational genes show the significance of complementarity between virus population components in viral infection.The work by Lu et al. looked into the impact of this genetic differentiation on the ability of the aphid to respond to unfavorable plants. Single nucleotide polymorphism (SNP) sites in the APV genomes of the pea aphid colonies in Vicia faba and Vicia villosa, respectively, were discovered using a transcriptome investigation. When host aphids were switched from high-fitness plants V. faba to low-fitness plants V. villosa or Medicago sativa, the SNP at site 5,990, G5990A, located in the RNA-dependent RNA polymerase (RdRp) domain, showed a change from G to A. In RdRp, this SNP resulted in the asparagine (N) to serine (S) substitution at site 196. S196N was located at a random coil distance from the conserved active motifs, and the N196 type of RdRp's polymerase performed 44.5% better than the S196 type. The increased APV replication rate was caused by the enhanced enzymatic activity of RdRp positively impacted insects by lowering plant defenses against aphids, and it sheds light on a scenario in which a host can benefit from modifications of a symbiotic virus when the host adapts to the changed environmental conditions.Host adaptation causes significant genetic changes in symbiotic microorganisms in insects; however, it is still unknown how virus mutations in symbiotic hosts affect host adaptability to new environments and increase viral fitness in hosts. Acyrthosiphonpisum virus (APV), a symbiotic virus, has undergone genetic divergence in one location, and Bemisia tabaci (Hemiptera: Aleyrodidae) is a highly efficient vector for the spread of the chili leaf curl virus , which poses a considerable obstacle to the cultivation of chili in South Asia. Nekkanti et al. investigated the molecular processes driving interconnections between B. tabaci and ChiLCV transmission and found potential alternative candidates for the control of the B. tabaci-begomovirus complex. Transcriptome analysis of B. tabaci revealed 80 genes with distinct expression patterns after 6 h of ChiLCV intake, with 29 being positively regulated and 51 being negatively regulated. The Kyoto Encyclopedia of Genes and Genomes (KEGG) research of the differentially expressed genes (DEGs) revealed their role in metabolic activities, signaling channels, cellular processes, and organismal systems. Reverse transcription quantitative real-time PCR (RT-qPCR) was used to confirm the expression of positively regulated genes following the acquisition of ChiLCV. DEGs that were abundant in B. tabaci and promoted viral infection and circulation included replication factor A protein 1, fasciclin 2, inhibin beta chain, cytosolic carboxypeptidase 3, dual-specificity protein phosphatase 10, 15, dynein axonemal heavy chain 17, and Tob1.Alternaria alternata apple pathotype is responsible for Alternaria leaf blotch, a serious fungal disease that reduces apple output worldwide. Interest in mycoviruses as a potential biocontrol agent has grown significantly due to their ability to impart hypovirulence on their hosts. Li et al. found that A. alternata f. sp. mali strain QY21 was coinfected with A. alternata chrysovirus 1 strain QY2 (AaCV1-QY2) and A. alternata magoulivirus 1 (AaMV1), which belong to the genera Betachrysovirus and Magoulvirus, respectively. Hypovirulence of A. alternata was associated with these two mycoviruses, with AaCV1-QY2 likely contributing significantly. AaCV1-QY2 can alone postpone development and lessen host virulence, as shown by the fact that the lack of AaMV1 in strain QY21 had no influence on the hypovirulence phenotype. Viruses also prevented strains of A. alternata from overproducing the mycotoxin alternariol. AaCV1-QY2/AaMV1 mycoviruses may infect diverse strains of A. alternata, which can increase the effectiveness of interspecific dissemination of AaCV1-QY2.Kumar et al. identified co-infections of tobamoviruses, frangipani mosaic virus (FrMV), and plumeria mosaic virus (PluMV), from the temple tree, or Plumeria rubra f. acutifolia. The existence of PluMV was also observed in association with the distinctive symptoms of Gomphrena globosa (globe amaranth), a non-host of FrMV. To propagate PluMV, simple rub-inoculation was quite successful, and tobacco, globe amaranth, brinjal, datura, and chili were able to discriminate PluMV from FrMV in a host range assessment. The full genome sequence of PluMV was analyzed, revealing the genomic structure characteristics of tobamoviruses encoding four proteins: small replicase, large replicase, movement protein, and coat protein. The genome of PluMV was longer than that of FrMV; it shared significant sequence similarities with FrMV and other tobamoviruses, and it had a close but diverging evolutionary relationship with FrMV. RT-PCR assays confirmed the natural occurrence of PluMV in temple tree species in India, either alone or in combination with FrMV. In the second research article related to the diagnosis of viruses, Hamim et al. reported that virus-infected leaf samples stored in RNAlater® were acceptable for RT-PCR, PCR, Sanger sequencing, high-throughput sequencing (HTS), and enzyme-linked immunosorbent assay (ELISA)-based diagnostic tests. Agricultural productivity is significantly impacted by viral plant diseases, and accurate detection and characterization of viral infections are crucial for crop disease management. In order to carry out diagnostic tests using sensitive molecular techniques, intact nucleic acids present in plant tissue samples obtained from remote areas are required. RNAlater® offers effective, trustworthy sample preservation by preserving both RNA and DNA in plant tissue samples. Hamim et al. used leaf tissue samples from agricultural fields in Bangladesh that had viral symptoms to assess the viability of this method. Their research suggests that RNAlater® technology could be employed to characterize viruses in samples that have been kept for a long period and obtained from distant areas. This strategy will allow underdeveloped countries with limited laboratory facilities to greatly improve their capacity to identify and control viral infections in agricultural plants using cutting-edge analytical methods.Two original research articles about the diagnosis and characterization of plant viruses are included in this Research Topic. In the first paper, Zhuang et al. analyzed these findings and classified the impacts on biological processes in accordance with the viral life cycle.A comprehensive review article on this Research Topic contributes to our understanding of the biological roles of phosphorylation in the interactions between plants and viruses. One of the most well researched post-translational modifications, phosphorylation regulates multiple cell signaling pathways and is crucial for modulating the viral infection cycle in plants. The molecular mechanisms underlying the phosphorylation of plant virus proteins have been the subject of a considerable number of studies. In conclusion, this Research Topic provides state-of-the-art research methods on virus interactions with different hosts in agricultural systems. We would like to take this opportunity to express our gratitude to each of the authors and reviewers, whose outstanding work enabled the publication of this Research Topic. We believe that this collection will increase knowledge, and awareness of the significance of viral diseases, enable better surveillance and potential control of emerging viral diseases in agricultural systems, and prevent epidemics in agricultural crops.IH wrote the manuscript. All authors contributed to the article and approved the submitted version.


A recently-surfaced virus called Monkeypox virus (MPXV) has gained widespread attention as it dominates the news, creating a sense of panic among people due to the potential threat it poses to their health.To evaluate knowledge about this virus and its disease, and to raise consciousness among the members of the Faculty of Natural and Life Sciences and Earth and Universe Sciences at the University of Tlemcen, we launched an online web-based survey for a twenty days’ period that contained sociodemographic and perceptiveness questions about the emergent virus, its disease, and vaccination.Our findings showed that the majority of the respondents of our study have a satisfactory level of knowledge about this emerging virus and its disease. Moreover, most participants showed a positive attitude towards the vaccine, considering it the best preventive means to fight against Monkeypox disease.Although the MPXV may not become a pandemic, but knowing the various ways that contribute to its spread is essential to avoid any possibility of a new one, especially in Algeria. The purpose of our study is to evaluate knowledge of teachers, administrative workers and students of the Faculty of Natural and Life Sciences and Earth and Universe Sciences of University of Tlemcen (NLS/EUS) about this contagious virus in order to promote awareness about the disease and its transmission for anticipating any outbreak in Algeria.The emergence and the growing number of Monkeypox virus (MPXV) cases worldwide pose a serious threat to human life, especially in non-endemic countries like Algeria. Additionally, the general poor public understanding of Monkeypox, which makes it a prerequisite for controlling and preventing this zoonotic virus and its disease  version 25 and Microsoft Office Excel 2007, as well as the creation of tables and graphs.In this section, results obtained from the survey were interpreted according to the following criteria.. These results may be explained by the fact that younger generations use more of social media than others, and more specifically, females that are interested in learning about the latest developments in the field of health.The results indicate that the majority of respondents were women and belonged the 21-40 years’ age group (46.4%) et al., 2022), Nigeria , China , and Jordan .Results show that most of the participants (90%) previously knew that this disease is related to a virus. These results are consistent with precedents studies carried out in Indonesia  also found the same result in their study from Lebanon. It could be explained by the fact that this virus caused several outbreaks in African and non-African countries so it becomes a topic of news in social media.Among the 125 participants, more than two-thirds (61%) have heard about the emergence of the new Monkeypox virus before completing this questionnaire, Youssef et al., 2022). As a result, all of these transmission routes may be present in combination, for that it is difficult to distinguish between them in any given individual exposure or outbreak scenario . In our study we found that among the 66 respondents who declared knowing the sources of transmission of the virus, the majority strongly agreed that Monkeypox could be transmitted through contact with an infected animal or person  .The answers of the respondents about the modes of the Monkeypox virus transmission show that just 47% of acknowledge knowing them. This relatively limited information can be explained by the fact that Algeria has not yet been exposed to an epidemic of this virus. This zoonotic virus can transmit through many routes . Historically, the virus did not spread efficiently between humans, requiring direct contact with infected body fluids or fomites (bed sheets) or sustained respiratory droplet exposure within 6 meters for 3 hours or more , but recently this emerging virus has proven its great ability to spread among people, and the biggest evidence was in May 2022 where more than over 17,300 confirmed and suspected cases were identified, and over 40.000 infected people in 87 countries that were not MPXV endemic .Almost half of participants (43%) did not know if MPXV does spread easily between people or not, and only 39% of them were aware that this virus does spread easily between people, which is in agreement with a previous study conducted in Saudi-Arabia in 2022, where only 41.9% of the participants unanimously agreed on the ease of its transmission between people . Besides, it is very important to note that homosexuality plays a major role in the emergence of this virus. The CDC Data suggest that most patients are amongst the homosexual or bisexual men in the current outbreak .According to the participants, there are several factors that promote their emergence and a large part of them (32.2%) consider that the decrease in vaccine protection against smallpox is the first responsible reason for this phenomenon et al., 2022 ; Harapan et al., 2022 ; Hraib et al., 2022 ; Titanji et al., 2022).Among the 24 respondents, they indicated that they may be exposed to this disease by carrying associated symptoms, like fever (39.5%), muscle pain and backache (23.2%), skin eruption (23.2%), and swollen lymph nodes (14%). This might be due to the high similarity between the symptoms of Monkeypox, Smallpox, Measles and Chickenpox which may also explain by the fact that Algeria has not yet recorded any official cases of this virus , and to maintain a distance of at least 1 meter from suspected individuals, as well as wearing a well-fitted mask and disposable gloves around them .The results show that 48% of the participants knew the preventive measures to avoid transmission of this disease. This result can indicate the consciousness of our participants to follow the news and be updated on the hygienic status concerning emerging viruses in order to avoid repeating the same mistakes made previously in the COVID-19 pandemic era. Whereas according to different studies, there are many preventive measures that can be considered to avoid MPX infection including, avoiding direct contact with animals that are suspected of harboring MPXV, especially in geographical locations where Monkeypox disease is prevalent, to avoid contact with any material that has been in contact with a sick animal or human . On top of that, those with suspected or confirmed MPV infection should isolate themselves at home or at another location covering all lesions, wear a mask around others, wash or disinfect their hands after touching lesions, and not share any personal items .Furthermore, cooking all foods containing animal meat or components thoroughly .In our study, 71 participants have declared that the best ways to prevent the transmission of this virus were to avoid contact with infected animals and sick people, wash hands frequently with soap and avoid illegitimate sexual relations  77.3% of the participants knew the therapeutic and preventive means against this disease, and most of them considered that a vaccine is the best way to prevent and treat this disease with 33.3%, followed by antiviral drugs (21.7%). To date, there are no specialized vaccines and antiviral drugs available for MPXV, although other previous vaccines for small Chickenpox viruses, Cidofovir, ST-246, and VIG can be used to control the Monkeypox epidemics . There are categories of people who are more concerned by this treatments including, those whom are suffering from serious diseases such as hemorrhagic diseases, confluent lesions, sepsis, encephalitis, or other diseases requiring hospitalization, also, the immunocompromised population, the pediatric population, especially those under 8 years old, without forgetting pregnant or breastfeeding women, those with a history of allergic dermatitis, and individuals with other active exfoliative skin diseases, in addition to those with one or more complications like, secondary bacterial skin infection ,bronchopneumonia, concurrent diseases, or other co morbidities .et al., 2021a,b ; Kaler et al., 2022). also as both of them share some common symptoms, like, fever, headache and vomiting ). But we also notice some differences between the two of them like the incubation period which is between 5 and 15 days for SARS-CoV-2 virus and between 5 to 3 weeks for Monkeypox virus, further, the difference in terms of the key symptom, like the classic rash of Monkeypox and ARDS (Acute Respiratory Distress Syndrome) for COVID-19 .Among all of our participants, 58% indicate that there are no similarities between the two infections, while the rest of them (42%) do distinguish that, there are some common ones . By comparing many studies, we notice that there are some similarities between the two infections in terms of methods of transmission, as both can be transmitted to people through close, direct contact, with infected person’s secretions and droplets, or in terms of the preventive measures that must be taken in order to limit their spread . Adding to that, the difficulty to eradicate this virus since it remerges and exists in multiple animal reservoirs . For that every moment there is a possibility and risk of exposure to this new pandemic.Our data shows that 52% of participants think that there is a risk of having a new pandemic like the one of SARS-CoV-2. However, the WHO director stated that the infectivity of MPXV is relatively low and this virus may not become a pandemic. Despite these allegations, MPX outbreak has become endemic in more than 20 countries since May 2022 (Gong In our study which aimed to evaluate knowledge of Teachers, Administrative-Workers and Students of the Faculty of Natural and Life Sciences and Earth and Universe Sciences of University of Tlemcen about Monkeypox virus in order to spread awareness about the disease and its transmission for anticipating any outbreak in Algeria, we found that our participants had a satisfactory level of knowledge for the mode of transmission, and the preventives measures to fight against the spread of this emerging virus. Furthermore, most of the participants showed a positive attitude towards vaccines, considering them as the best therapeutic means to combat the spread of MPX disease. Although there are still many unanswered questions about Monkeypox disease, animal reservoirs, and the virus itself, through this humble work we tried to answer and clarify some important points that all people should know about emerging viruses in general and Monkeypox virus and its disease in particular, in order to prepare in advance for any outbreaks in Algeria.The data that support the findings of this study are available from the corresponding author upon request.The authors declare no conflict of interest.MPXVMonkeypox VirusMPXMonkeypox diseaseARDSAcute Respiratory Distress SyndromeCOVID-19Coronavirus Disease 2019SARS-CoV-2Severe Acute Respiratory Syndrome Coronavirus 2NLS/EUSNatural and Life Sciences and Earth and Universe Sciences


Mops condylurus bats in Kenya, indicating the virus is well-tolerated. Our findings indicate M. condylurus bats may be a reservoir host for Bombali virus. Increased surveillance of these bats will be important to reduce potential virus spread. We found similar mild perivascular inflammation in lungs of Bombali virus–positive and –negative  Despite extensive research since the first documented human Ebola virus (EBOV) disease outbreak in 1976, animal species involved and mechanisms by which ebolaviruses spillover to humans remains enigmatic. Bats have been implicated as reservoir hosts , the sixth and most recent ebolavirus identified. First reported in M. condylurus bats; the BOMV-negative bats were used as negative controls  as described previously  support bats as a chronic subclinical BOMV reservoir with the potential for infection and sporadic virus excretion. However, the small number of samples, absence of recovery of infectious virus, and lack of longitudinal data make interpretation of our results difficult. As for all wildlife-borne ebolaviruses, no direct evidence currently exists regarding the transmission route of BOMV between susceptible hosts. Our findings suggest the mode of transmission involves activation of replication in either macrophages or other cells and virus dispersal through saliva droplets or feces. The identification of BOMV infection in lungs and occasional virus presence in intestines further supports this hypothesis (M. condylurus bats and an absence of substantial concurrent pathologic findings. Our results support M. condylurus bats as reservoir hosts for ebolaviruses and highlight the importance of bat surveillance to understand and mitigate potential emerging disease risks.In summary, we showed EBOV-VP40 immunolabeling, acting as a cross-reactive surrogate for BOMV antigen, in pulmonary macrophages of wild BOMV RT-PCR–positive Mops condylurus bats naturally infected with Bombali virus, Kenya. Additional information for no substantial histopathologic changes in


Efficient spread of respiratory viruses requires the virus to maintain infectivity in the environment. Environmental stability of viruses can be influenced by many factors, including temperature and humidity. Our study measured the impact of initial droplet volume  and relative humidity  on the stability of influenza A virus, bacteriophage Phi6 (a common surrogate for enveloped viruses), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) under a limited set of conditions. Our data suggest that the drying time required for the droplets to reach quasi-equilibrium  varied with RH and initial droplet volume. The macroscale physical characteristics of the droplets at quasi-equilibrium varied with RH but not with the initial droplet volume. Virus decay rates differed between the wet phase, while the droplets were still evaporating, and the dry phase. For Phi6, decay was faster in the wet phase than in the dry phase under most conditions. For H1N1pdm09, decay rates between the two phases were distinct and initial droplet volume had an effect on virus viability within 2 h. Importantly, we observed differences in virus decay characteristics by droplet size and virus. In general, influenza virus and SARS-CoV-2 decayed similarly, whereas Phi6 decayed more rapidly under certain conditions. Overall, this study suggests that virus decay in media is related to the extent of droplet evaporation, which is controlled by RH. Importantly, accurate assessment of transmission risk requires the use of physiologically relevant droplet volumes and careful consideration of the use of surrogates. Respiratory viruses, such as influenza A virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), contribute to high morbidity and mortality. These viruses must remain infectious in the environment for transmission to the next host to succeed. Understanding how environmental, host, and virus factors impact the stability of expelled virus will lead to a better assessment of virus transmission risk and ways to reduce it.Many factors can impact virus stability in the environment, including virion structure, temperature, relative humidity (RH), droplet composition, solute concentration, and fomite surface material –5. Howev–Studies measuring virus stability in the environment typically use one of two methods to generate droplets: nebulizers to produce aerosols or pipettes to create droplets with as much as 50 μL per droplet. While large droplets are commonly used to assess environmental virus stability, they do not mimic the physiological volume of a droplet created by an expulsion. The vast majority of expelled droplets from the respiratory tract are less than 0.5 μL in volume (approximately 1 mm in diameter for a sphere); a droplet of 50 μL (approximately 4.6 mm in diameter for a sphere) is about 5 times larger and 100 times greater in volume . StudiesThis study primarily used the 2009 pandemic influenza H1N1 virus  and bacteriophage Phi6, a commonly used virus surrogate, to examine environmental stability of enveloped viruses in three different droplet volumes at three different RHs over time. Specifically, we measured the viability of each virus in 50-, 5-, and 1-μL droplets on surfaces over time at 40%, 65%, and 85% RH. We observed that virus within smaller droplets decays quickly regardless of RH, while virus decay occurs more slowly in larger droplets. We also explored droplet evaporation rates and found that virus decay is closely correlated with the extent of evaporation, which is likely a proxy for the solute concentration in the droplet. Additionally, limited experiments with SARS-CoV-2 showed that influenza virus decayed similarly to SARS-CoV-2 at an intermediate (55 to 60%) RH in 50-, 5-, and 1-μL droplets. Overall, our results suggest that virus stability studies should use smaller, more physiologically relevant droplet volumes and should recognize the limitations of surrogate viruses.https://doi.org/10.6084/m9.figshare.21711119, https://doi.org/10.6084/m9.figshare.21711122, and https://doi.org/10.6084/m9.figshare.21711116). These RHs were selected to match values in other published work  for droplets to reach quasi-equilibrium at each RH. Droplet mass was measured on a microbalance in an environmental chamber and recorded every minute. Download FIG S1, PDF file, 0.1 MB.Initial droplet volume impacts drying kinetics. (A to C) Mass normalized to starting mass for all droplet volumes at 40% (A), 65% (B), or 85% (C) RH over time. (D) Summary data showing the time  were weighed over time. The time of quasi-equilibrium was determined and the mass at quasi-equilibrium was measured. Download Copyright © 2023 French et al.2023French et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the To directly examine how RH and droplet volume impact virus stability in the environment, we applied virus in droplets of different volumes to a polystyrene surface and quantified recovery of infectious virus over time . We comp10.1128/mbio.03452-22.2FIG S2https://doi.org/10.1056/NEJMc2004973) are unknown. Download FIG S2, PDF file, 0.4 MB.Environmental conditions of H1N1pdm09 droplets and Phi6 droplets were within 5% of targeted RH and maintained temperatures between 20 and 28°C. The RH and temperature of the environmental chamber were recorded every 15 min during H1N1pdm09 stability experiments at (A) 40% (A), 65% (B), and 85% (C) RH. The RH and temperature of the environmental chamber were recorded every minute during Phi6 stability experiments at 40% (D), 65% (E), and 85% (F) RH. The RH and temperature data for comparing decay of H1N1pdm09 at 60% (G) with SARS-CoV-2 at 55% (H) were recorded every 15 min or 1 min, respectively. Light green shows the temperature at each replicate, and dark green indicates the average temperature for the 3 independent replicates. Light blue shows the RH for each replicate, and dark blue indicates the average RH for the 3 independent replicates. (I) Absolute humidities were calculated for all experiments. The keys show the corresponding average RH for each condition instead of targeted RH. Conditions for data previously published by van Doremalen et al. /mL for SARS-CoV-2 and our viruses were 106 PFU or TCID50/mL, we believe this difference to be negligible  droplets than in small (1-μL) droplets, that decay rates during the wet phase are greater than or similar to decay rates during the dry phase regardless of droplet size and RH, and that differences in virus decay are more common in 50-μL droplets than in 1-μL droplets and at low RH.Our results raise questions about the relevance of prior studies on stability of viruses that employ large droplet volumes. For example, one study derived a half-life (~0.3-log decay) of 6.8 h for SARS-CoV-2 in 50-μL droplets on polypropylene plastic  9). The. The9). We have attempted to study viruses in a more realistic droplet volume than those used in past research, but even a 1-μL droplet is at the extremely large end of the range of droplet volumes observed in respiratory emissions. During talking, coughing, and sneezing, droplets of this volume are emitted in much lower numbers, by many orders of magnitude, than those that are 100 to 1,000 times smaller in diameter and that behave as aerosols . While wOur results also suggest caution in the use of surrogates to study the stability of pathogenic viruses and their potential for transmission. Surrogates that require fewer biosafety precautions are attractive for obvious reasons . They caOne limitation of this study is that we investigated virus persistence in culture medium, DMEM, that may not be representative of real respiratory fluid. We chose to use this medium for the purpose of comparing results with prior studies of SARS-CoV-2 in DMEM . Prior sExtrapolating our results to smaller droplet sizes and combining them with the findings of other studies may provide mechanistic insight into the dynamics of virus inactivation in droplets and aerosols. The biphasic virus decay that is readily observed in droplets likely occurs in aerosols, too , 25, 26.Although virus stability in droplets and aerosols appears to be a complex function of droplet size, composition, humidity, and other variables, mechanistically their role is to modulate the microenvironment surrounding a virion , 26, 27.10.1128/mbio.03452-22.3FIG S310 virus decay was plotted against percent original mass for Phi6, H1N1pdm09, and SARS-CoV-2 in 1 × 50 μL, 5 × 5 μL, and 10 × 1 μL at 40% RH, 65% RH, and 85% RH. Percent original mass was determined by determining droplet mass at 0, 20, and 40 minutes, then 1, 4, 8, and 24 h. RH listed in the legend show the targeted RH. Actual RH data are available in https://doi.org/10.1056/NEJMc2004973), for which actual RH values are unavailable. Download FIG S3, PDF file, 0.1 MB.Evaporation is major determinant of virus decay regardless of initial droplet volume. LogCopyright © 2023 French et al.2023French et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mbio.03452-22.10TABLE S7Table S7, PDF file, 0.1 MB.Genome copies for limited SARS-CoV-2 and H1N1pdm09 experiments demonstrate consistent recovery and genome stability. Download Copyright © 2023 French et al.2023French et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Due to our findings on the sensitivity of virus persistence to both droplet volume and composition, we urge a shift toward the use of more realistic conditions in future studies. They should employ droplets as close in volume as possible to those released from the respiratory tract  and whose chemical composition closely mimics that of real respiratory fluid. These findings are critical for pandemic risk assessment of emerging pathogens and useful to improve public policy on optimal transmission mitigation strategies.2 in minimum essential medium containing 10% fetal bovine serum (FBS), penicillin-streptomycin, and l-glutamine. Influenza A virus A/CA/07/2009 was derived from reverse genetics and grown in MDCK cells for 48 h in DMEM  containing 2% FBS, antibiotic-antimycotic, and l-glutamine. Stocks were diluted to 106 TCID50/mL using DMEM containing 2% FBS, penicillin-streptomycin, and l-glutamine for use in experiments. Virus titers were measured using the 50% tissue culture infectious dose assay on MDCK cells and calculated using the Spearman-Karber method (MDCK cells (obtained from the ATCC) were grown at 37°C in 5% COr method .6 PFU/mL for use in experiments. Virus titers were quantified by plaque assay  containing 2% FBS, antibiotic-antimycotic, and l-glutamine for use in experiments. All SARS-CoV-2-related work was conducted in biosafety level 3 (BSL3) laboratories.SARS-CoV-2 strain USA-WA1/2020  was passaged in Vero cells once before receipt and subsequently in Vero cells once upon receipt. Vero cells were grown in DMEM  supplemented with 5% FBS , 100 units/mL of penicillin, and 100 mg/mL of streptomycin  and maintained at 37°C and 5% COeviously . Stocks 50 and plaque assay have a linear relationship, virus decay should be comparable between virus quantification methods  placed in the environmental chamber.3, respectively  at room temperature and three  or four  RHs. A logger (HOBO UX100-011) placed inside the chamber recorded temperature and relative humidity. The temperature and humidity for all experiments are presented in ectively . DropletWe measured the stability of SARS-CoV-2 in an airtight desiccator at room temperature and 55% RH as described previously . The meaAll H1N1pdm09 and Phi6 collections were performed in technical duplicates and independent triplicates. Due to biosafety constraints and the additional resources required, the SARS-CoV-2 work was performed in technical duplicates with two independent replicates. Data from all replicates are presented for each virus.m(0)] immediately after the droplets were deposited onto the polystyrene surface. We calculated the percent original mass of the droplet over time using the following equation:We measured the initial mass -TCAGGCCCCCTCAAAGCCGA-3BHQ1-3′). In vitro-transcribed RNA served as standards to determine genome copies. Each qPCR plate contained duplicates of standards, samples, and no-template controls.We performed reverse transcription-quantitative PCR (qRT-PCR) on one independent replicate each of H1N1pdm09 and SARS-CoV-2 to address genome instability as a possible mechanism of inactivation. For SARS-CoV-2 the Qiagen QIAamp Viral RNA minikit was used to isolate RNA according to the manufacturer’s protocol. The 2019-nCoV RUO primer/probe kit targeting the N1 gene (IDT) was used in combination with the iTaq universal probe one-step kit. Synthetic SARS-CoV-2 RNA (BEI Resources) was used as a standard. For H1N1pdm09, the Qiagen QIAamp viral RNA minikit was used to isolate RNA according to the manufacturer’s protocol. The influenza virus M gene was targeted using the iTaq universal probe one-step kit with primers  for Phi6 wet-phase decay, but there was statistically significant variance for H1N1pdm09 wet-phase decay (P = 0.037), indicating that there is an interaction between initial droplet volume and RH that affects wet-phase decay.We conducted a multivariate analysis of variance (MANOVA) to determine if there was an interaction between initial droplet volume and RH that would account for differences in the wet phase decay of the viruses. There was no statistically significant amount of variance (https://doi.org/10.6084/m9.figshare.c.6458767. In addition, movies corresponding to the droplet drying over time at 3 different relative humidities can be found within the collection and at these individual links: https://doi.org/10.6084/m9.figshare.21711119, https://doi.org/10.6084/m9.figshare.21711122, and https://doi.org/10.6084/m9.figshare.21711116.All raw data associated with this study are available on Figshare at


We then developed an innovative and reproducible approach based on finger-pad transfer to evaluate the antiviral activity of HITS against a murine norovirus inoculum under conditions closely reflecting real-life surface exposure.Environmental surfaces, including high-touch surfaces (HITS), bear a high risk of becoming fomites and can participate in viral dissemination through contact and transmission to other persons, due to the capacity of viruses to persist on such contaminated surface before being transferred to hands or other supports at sufficient concentration to initiate infection through direct contact. Interest in the development of self-decontaminating materials as additional safety measures towards preventing viral infectious disease transmission has been growing. Active materials are expected to reduce the viral charge on surfaces over time and consequently limit viral transmission capacity through direct contact. In this study, we compared antiviral activities obtained using three different experimental procedures by assessing the survival of an enveloped virus (influenza virus) and non-enveloped virus  over time on a reference surface and three active materials. Our data show that experimental test conditions can have a substantial impact of over 1 log Several reviews and models have suggested that indirect contact transmission involving contaminated surfaces (fomites) could be the predominant transmission route for certain respiratory and enteric viruses ,2. In fanfection ,9. Such nfection ,11,12. Lnfection ,14 provinfection . The potnfection ,17. StudIn recent years, growing interest in the use of self-decontaminating surface materials as additional safety measures towards preventing disease transmission has emerged . These m10 TCID50/PFU per cm2) of active materials corresponds to the log10 difference between the infectious titer of a virus found on an untreated product versus an antiviral-treated product after inoculation or contact with viruses during a defined period. Among the limited number of experimental procedures reported, the international standard ISO21702, the counterpart standard of the ISO22196 for antibacterial activity, makes it possible to determine antiviral activity of an active surface onto which a viral inoculum in liquid state has been deposited for a defined contact period at 25 °C and at 90% RH [The antiviral activity  and nonenveloped virus  over time on a reference surface and three active materials. We then developed an innovative approach based on finger-pad transfer to evaluate the antiviral activity of high-touch active materials against a murine norovirus inoculum, a surrogate for human gastroenteritis viruses, under conditions closely reflecting real-life surface exposure.Murine norovirus , feline calicivirus  and human influenza virus  were produced on specific cell lines: RAW 264.7 (ATCC TIB-71) for MNV-1, KE-R (CCLV-RIE 0138) for FCV and MDCK (ATCC CCL-34) for H3N2 virus. Cells were cultivated in DMEM 1 g/L, DMEM 4.5 g/L or in EMEM supplemented with L-glutamine, antibiotics, and a controlled quantity of FCS (10% or 5%). Virus production and quantification were performed in an infection medium (EMEM or DMEM with 2% or without FCS). For influenza virus, Trypsin acetylated from bovine pancreas at 2 µg/mL was added to the culture medium.®, MetalSkin Technologies, Balaruc les Bains, France).Stainless steel discs  were used as a reference surface for both antiviral activity and fingerprint experiment. Stainless steel discs were functionalized, which included a DECON 5% bath and a 70% isopropanol bath interspersed with water bath. Stainless steel discs were sterilized by autoclaving for 15 min at 121 °C. AS1 material corresponds to a PVC membrane with a topcoat layer (PMMA acrylic and PVC resins) containing micro-silver glass phosphate particles (<15%). AS2 material corresponds to the 1.4 mm layer obtained by extraction, containing 82–87% PVC resin, 0.8–1% silver, 1–2% Ca/Zn heat stabilizer and 12–15% of additives . The AS3 material corresponds to a 120 µm layer of a polyester/metal mix (90–93% copper) sprayed over stainless steel coupons  and the Spearman–Kärber technique. The adapted protocol (ISO21702AD) corresponds to the ISO21702 protocol, except that the inoculum was not covered by a film and the RH during incubation period was 41%. The NFS90700AD protocol consisted of depositing 20 µL (1.107 TCID50/mL) onto a 1 cm2 surface [50) and the Spearman–Kärber technique.Three protocols for the determination of antiviral activity were compared in this study. The first one corresponds to the ISO21702 standard . A volum surface . The ino2 area of a middle fingertip previously cut from a nitrile glove and put under hood flow for quick drying by streaking the inoculum. Once dry, the fingertips were positioned onto the middle finger of a gloved hand of an operator and pressed on to the test surfaces for 10 s with one rotating movement of the finger (90°) and a mechanical pressure of 1 kg controlled using a scale. The surfaces where the fingertips made contact were washed in maintenance medium and the virus remaining on contaminated nitrile-gloved middle fingertips was recovered by sonication procedure . The samples were then prepared for quantification utilizing tissue-culture infectious dose 50% (TCID50). The residual virus was quantified by Spearman–Kärber methods. For the deposit of complex inoculum, 20 µL of mixture (V/V) containing murine norovirus (1.33.107 TCID50/mL) and Enterococcus faecium bacteria (4.56.107 CFU/mL) was deposited onto a 1 cm2 area. The surfaces where the fingertips made contact were washed in 2 mL of maintenance medium. Half of the volume was used for bacterial quantification on Tryptone soy agar . The CFU were quantified after 24–48 h at 37 °C. The other half of the volume was centrifuged to eliminate the bacteria and used for viral quantification using tissue-culture infectious dose 50% (TCID50).A volume of 20 µL of viral suspension (1.107 TCID50/mL) was deposited onto 1 cm5 TCID50) onto a 16 cm2 area, then left the inoculated surfaces in an incubator at 25 °C, half with a RH of 90%  and the other half at a RH of 50% (ISO21702AD protocol). At 90% RH, the inoculum remained in a liquid state throughout the incubation period, while the viral inoculum completely dried after an incubation period of 5 to 6 h post application at 50% RH (data not shown). We also prepared a second group of inoculated supports according to the NFS90700 modified protocol (NFS90700AD), corresponding to the deposition of 20 µL of viral solution (2.10 × 105 TCID50) onto a 1 cm2 surface area with drying through mechanical contact with pipette tips, allowing the complete drying of the inoculum in 5 min post application. Inoculated supports were then left in an incubator at 20 °C with an RH of 50% (NFS90700AD protocol). For each test surface and experimental condition, inoculated supports were retrieved at desired time points  and immediately soaked with viral culture media before determining the infectious viral titer (TCID50) (Spearman and Kärber Methods). To compare viral persistence on stainless steel using different experimental conditions, biphasic linear regression plots (log10 titer vs. time) of the survival data were used to calculate the half-life , the D value  and the residual number of infectious viruses on the support at 5, 15, 30 and 60 min and 2, 8, 24 and 48 hours post application. We reported all experimental measurements as means of 18 replicates with SD. A Student statistical test with a confidence interval of 95% was carried out to evaluate significant differences among the data .We aimed to investigate the impact of different experimental parameters used in three antiviral activity protocols , including virus deposition- and environmental-specific parameters, on viral persistence on a particular surface. To this end, we first tested the impact of these parameters on the survival of both enveloped H3N2 influenza virus and the non-enveloped feline calicivirus on stainless steel surfaces, used as the reference support to determine the antiviral activity of active material over a defined period of time. We prepared one group of inoculated supports according to the ISO21702 protocol, corresponding to the deposition of 400 µL of viral solution . For H3N2, among all experimental parameters from the ISO21702 and ISO21702AD protocols, only the amount of virus at 24 h post application was found to be significantly different (p = 0.039). However, the amount of virus at 8 h (p = 0.005) and 24 h (p = 0.005) post application, half-life (p = 0.001) and D (p = 0.001) values were significantly different between ISO21702 and NFS90700AD protocols (p = 0.01), were significantly different between ISO21702AD and NFS90700AD protocols. As observed for H3N2 virus, feline calicivirus showed high stability with no significant differences among the three experimental procedures at 60 min post application (p = 0.005) and 24 h (p = 0.036) post application was significantly different between the two protocols, associated with an average loss of 1.6 log10 TCID50 at 48 h post application (p < 0.001/p < 0.0001), 8 h (p < 0.0001/p < 0.0001), 24 h (p = 0.005/p < 0.0001) and 48 h (p < 0.001/p = 0.013) post application, half-life (p = 0.001/p = 0.018) and D (p = 0.001/p = 0.006). A further loss of around 0.8 log10 TCID50 was observed at 48 h post application using the NFS90700AD protocol as compared to the other two protocols  was determined. The average antiviral activity Rm (log10 TCID50/cm2), including a 95% confidence interval (Kr) of the three active materials from three independent experiments, was then determined. Antiviral activity R corresponded to the difference in the logarithm of the infectivity titer of the virus found on an untreated product and an antiviral-treated product after a period of incubation and contact with the virus. Stainless steel supports were taken as the untreated (reference) surface.We next evaluated the amount of residual H3N2 or feline calicivirus infectious viruses on each of three inoculated active materials  over time . As desc10 TCID50 was observed on AS03 at 15 min post application, between NFS90700AD and ISO21702 or ISO21702AD protocols (10 TCID50/cm2) of the three active surfaces, corresponding to the difference in the logarithm of the infectivity titer of H3N2 virus found on stainless steel supports versus antiviral-treated, were determined over time. While the antiviral activity of AS01 at 2 h post application showed no significant difference among the three protocols, antiviral activities were significantly higher at 8 and 24 h post application using the NFS90700AD compared to the ISO21702/ISO21702AD protocols  was quickly dried onto a 1 cm2 surface of the middle finger cut from a nitrile glove. Once dry, the contaminated fingertip was positioned onto the middle finger of a gloved hand of an operator and pressed onto the test surfaces for 10 s with one rotating movement of the finger (90°), associated with a mechanical pression of 1 kg at 20 °C and 40–60% RH. The surface where the fingertip made contact was washed in maintenance medium, and the residual amount of virus on the contaminated nitrile-glove fingertip was recovered by a sonication procedure. The samples were then prepared for the quantification of infectious viral titer (TCID50).To better understand if virus transfer can occur from the touching of a contaminated fomite, we next developed an experimental procedure adapted from the finger-pad transfer experiment proposed by Bidawid et al.  to enablWe first assessed the possibility of transferring murine norovirus (MNV-1), a surrogate for human gastroenteritis viruses, from gloved fingertips to a reference stainless steel surface . A soili10 TCID50 and 1.76 ± 0.18 log10 TCID50, respectively; the amount of virus isolated from the reference surface after the third contact was below the lower limit of quantification, but still detectable on the glove itself  was also evaluated  and Enterococcus faecium (105 PFU) in the presence of 3 g/L of BSA, from gloved fingertips to both reference stainless steel and AS03 supports  (The average amount of infectious norovirus isolated from the reference surface inoculated through contact with a gloved finger contaminated with the complex mixture was 3.6.105 log10) a. This c5 log10) a.10 TCID50 was observed at 5 min following the transfer onto the stainless steel supports in the presence of bacteria as compared to the virus alone (p = 0.01). The virus survival profiles obtained from the two experimental conditions then become very similar over time, up to 60 min post application when the residual amount of virus on the reference surface was significantly greater in the presence of bacteria (p = 0.001). The average amount of Enterococcus faecium bacteria isolated from the inoculated surface after complex-mixture-contaminated gloved-finger contact was 4.2×103 CFU (3.62 ± 0.21 log10)  c, correstransfer d.Enterococcus faecium over time after a finger-pad transfer onto AS03 active material  and 15 (p = 0.002) minutes post transfer between experimental conditions with or without bacteria (10 TCID50/cm2) at 5 min post transfer was not significantly different with or without bacteria. Similarly, the residual quantity of Enterococcus faecium bacteria after a finger-pad contact on the AS03 material decreased over time, almost reaching the LLOQ at 15 min post transfer , the amount of virus deposited per cm2 (5.30 vs. 4.14 log10 TCID50), and the relative humidity during the incubation period (50 vs. 90%). While the three experimental procedures had only a moderate impact on virus survival on the stainless steel support over time, observed differences were found to mainly depend on the viral strain tested. As expected, differences in residual virus quantity on the stainless steel support between the enveloped influenza virus and the non-enveloped calicivirus were apparent at 48 h post application. This is in accordance with previous findings of infectious enveloped viruses persisting for up to 5–7 days, and non-enveloped viruses for up to several weeks, on various smooth surfaces [2 compared to gentle drying on 16 cm2 inoculum according to the ISO21702AD protocol. To date, no study has been performed describing the impact of inoculum size (surface area and quantity) on viral survival on smooth surfaces. Interestingly, virus survival profiles over time are similar between ISO21702 and ISO21702AD experimental conditions, except for the residual quantity of the virus at 8 h post application for calicivirus and at 24 h post application for both H3N2 and calicivirus. Multiple mechanisms relating to the level of RH (50 vs. 90%), including the desiccation and interaction of viral capsids at the air-water interface (AWI) of a solution, may contribute to viral inactivation on surfaces [3 plaque-forming units (PFU) applied to a 1 cm2 coupon occurred in less than 60 min under dry conditions [2 than when the same quantity is spread over 16 cm2 or maintained in a liquid state. The antiviral activities of AS01 and AS02 were found to be stronger using the ISO21702 method against feline calicivirus compared to those measured using the NFS90700AD experimental procedure. The observed differences between the two protocols could be due to several factors, including the loss of virus on the reference surface and the potentially stronger action of ions released in the liquid inoculum on non-enveloped viruses [2) on the antiviral activity, with the viruses being in contact with a greater quantity of active elements. As observed for the enveloped virus, the NFS90700AD protocol enabled the strongest antiviral activity of the copper alloy in AS03 against the non-enveloped calicivirus, compared to the other two protocols. To our knowledge, our study is the first aimed at directly comparing different standardized or modified experimental procedures to evaluate the antiviral activity of active materials. Our data show that experimental test conditions can have a substantial impact of over 1 log10 on the antiviral activity of the active material for the same contact period. The observed impact of these protocols on the antiviral activities directed towards enveloped and non-enveloped viruses should now be confirmed on a large panel of active materials and using several other viral strains of interest. Projections of biological fluids from an infected individual onto surrounding surfaces are generally small in terms of volume, tending to dry quickly. Considering that the antiviral action of self-decontaminating surface material depends, in real-life conditions, on micro-nanomaterial/virus interactions occurring within a dry inoculum, the NFS90700AD protocol thus seems the most relevant experimental approach to evaluate antiviral activity of an active material.The role played by contaminated surfaces in the transfer and spread of viruses is now well established ,6. Theirsurfaces ,43. The surfaces ,46. The surfaces . Considesurfaces ,23. Durisurfaces ,23,47. Anditions . The ant5 TCID50 norovirus, a concentration plausibly present on the finger of a contaminated person. We transferred the virus efficiently to both the reference surface and the copper alloy active material representing the HITS. The use of the contaminated gloved finger-pad allowed the transfer of around 104 infectious norovirus particles onto the surfaces, corresponding to a finger/surface transfer rate of 4.4%, slightly lower than the 13% reported by Bidawid et al. with calicivirus [2 infectious norovirus particles remained present on the recipient reference surface at 60 min post transfer. The minimal dose of norovirus required to cause human infection could be between 10 and 100 infectious viral units [10 loss in the first 5 min post transfer, reaching the lower limit of quantification at 15 min post application. Considering the hypothesis of a 5% transfer efficiency of the virus through finger contact with a contaminated surface, the residual amount of norovirus on the active surface at 5 min post transfer was insufficient to allow the spread of the disease by finger contact. Thus, the theoretical transfer of the same quantity of virus, such as remains on a stainless-steel surface at 15 min post transfer, from AS03 onto a finger is only possible from 0 to 5 min post transfer. By comparing the amount of infectious virus on both reference steel and active surfaces, this experimental approach enables the determination of the antiviral activity of HITS that have been contaminated by contact under conditions closely resembling real life situations. The transfer via this approach of a mixture of virus and bacteria in soiled inoculum onto different surfaces has allowed the evaluation of antiviral activity of active HITS contaminated by a complex artificial mixture likely mimicking infectious microbial-containing fecal deposits transferred from contact with a soiled finger of an infected individual. Interestingly, the presence of the bacteria Enterococcus faecium present in human feces and used as a model in our study seemed to participate in the protection of norovirus by reducing the desiccation effect observed early on stainless steel supports. The protective role of bacteria on viruses has previously been described by Lopez et al. [Laboratory simulations have demonstrated that hand-to-hand and fomite-to-hand contact are plausible modes of transmission for enteric viruses provided that the viral particles remain viable on hands and fomites in a sufficient quantity to reach the digestive tracts via finger contamination ,14. Whilal units . In a woStreptococcus pneumoniae by finger-surface contact using the proposed approach and we are developing an innovative technology combining autofluorescent viruses and living human skin explants for the evaluation of antiviral activity of active materials.The experimental procedure proposed in our study and others offer the possibility of proposing a standardizable approach allowing the reproducible contamination of HITS in conditions resembling real-life via the transfer of infectious pathogens by finger contact. In response to the strong rise in the development of innovative self-decontaminating surface materials and in their promotion as a complementary strategy for disinfection to reduce the spread of microorganisms, our experimental approach should help meet the needs of production to test new surfaces in accordance with stringent end-user needs and requirements. We are currently studying the transfer of a mixture comprising enveloped respiratory virus (SARS-CoV-2 and influenza virus)/mucus nasal/saliva in the presence or not of


Rousettus aegyptiacus) and other Chiroptera species. The disease progression involves early viral replication impacting immune cells like monocytes, macrophages, and dendritic cells, followed by damage to the spleen, liver, and secondary lymphoid organs. Subsequent spread occurs to hepatocytes, endothelial cells, fibroblasts, and epithelial cells. MV can evade host immune response by inhibiting interferon type I (IFN-1) synthesis. This comprehensive investigation aims to enhance understanding of pathophysiology, cellular tropism, and injury sites in the host, aiding insights into MVD causes. Clinical data and treatments are discussed, albeit current methods to halt MVD outbreaks remain elusive. By elucidating MV infection’s history and mechanisms, this review seeks to advance MV disease treatment, drug development, and vaccine creation. The World Health Organization (WHO) considers MV a high-concern filovirus causing severe and fatal hemorrhagic fever, with a death rate ranging from 24 to 88%. The virus often spreads through contact with infected individuals, originating from animals. Visitors to bat habitats like caves or mines face higher risk. We tailored this search strategy for four databases: Scopus, Web of Science, Google Scholar, and PubMed. we primarily utilized search terms such as “Marburg virus,” “Epidemiology,” “Vaccine,” “Outbreak,” and “Transmission.” To enhance comprehension of the virus and associated disease, this summary offers a comprehensive overview of MV outbreaks, pathophysiology, and management strategies. Continued research and learning hold promise for preventing and controlling future MVD outbreaks.The Marburg virus (MV), identified in 1967, has caused deadly outbreaks worldwide, the mortality rate of Marburg virus disease (MVD) varies depending on the outbreak and virus strain, but the average case fatality rate is around 50%. However, case fatality rates have varied from 24 to 88% in past outbreaks depending on virus strain and case management. Designated a priority pathogen by the National Institute of Allergy and Infectious Diseases (NIAID), MV induces hemorrhagic fever, organ failure, and coagulation issues in both humans and non-human primates. This review presents an extensive exploration of MVD outbreak evolution, virus structure, and genome, as well as the sources and transmission routes of MV, including human-to-human spread and involvement of natural hosts such as the Egyptian fruit bat ( Recent cases in Guinea, Ghana, and the ongoing Equatorial Guinea outbreak accentuate the necessity for continuous surveillance and containment. The latter reported a swift succession of deaths, and Tanzania also reported fatalities attributed to the virus, highlighting the ongoing threat  and human-to-human transmission parallel other Ebola viruses, such as Sudan virus, Bundibugyo virus, and EBOV. Ongoing research has elucidated MV’s natural sources, including Hipposideros caffer, in tandem with Rousettus aegyptiacus (The Marburg virus (MV) has emerged as a significant global threat, instigating multiple fatal outbreaks with a considerable mortality rate since its discovery and characterization in 1967. Its initial appearance spanned regions including Marburg, Frankfurt (Germany), Yugoslavia (now Serbia), and Belgrade, with Africa primarily bearing the brunt of its impact . Accordig threat . MVD’s ig threat . As a me species . Its zooyptiacus . With anyptiacus . This re2.The Marburg virus (MV) exhibits an enveloped and pleomorphic structure, displaying uniform diameter but variable length filamentous, non-segmented, rod-like, cobra-like, circular/annular, and branched particles . Its vir3.3.1.Chlorocebusaethiops), was brought on by these imports. According to grivets’ reports, staff employees were primarily exposed to these sick wild animals’ meat and organs, which caused them to contract MV  and explored several bat-inhabited caves. After his return to Johannesburg, he exhibited symptoms such as fever, headache, myalgia, and vomiting, ultimately succumbing on 5 February 1975. His travel companion and a nurse who cared for him were secondary cases, both of whom survived through supportive treatment. Swift case isolation and contact tracing successfully contained the outbreak. While the source of infection remained unconfirmed, suspicion centered on the index case acquiring the virus through exposure to bats or their droppings within the caves .3.3.In 1980, Kenya experienced the third documented outbreak of Marburg-virus disease. The initial patient contracted the infection in western Kenya, leading to the subsequent infection of a doctor in Nairobi who had close contact with the patient, ultimately resulting in severe haematemesis. However, no additional instances of transmission within medical settings were observed. Surveillance efforts in western Kenya did not uncover evidence of Marburg-virus disease, but they did indicate the potential existence of Ebola hemorrhagic fever .3.4.In 1987, a single case of MVD outbreak emerged in Kenya, centering around a 15-year-old Danish boy who fell victim to the infection and subsequently passed away. The boy had encountered the virus within a cave inhabited by Egyptian fruit bats. This marked the inaugural documented instance of Ravn virus transmission, a near kin of the Marburg virus responsible for MVD. Effective containment measures were enacted through patient isolation and contact tracing, successfully preventing the emergence of secondary cases .3.5.In both 1998 and 2000, there was a single MVD outbreak situated in Durba, Democratic Republic of the Congo (DRC). The affected individuals were gold miners who toiled within a mine known to be home to Egyptian fruit bats, the virus’s natural reservoir. This outbreak encompassed a total of 154 cases and tragically led to 128 fatalities, resulting in an 83% case fatality rate. Remarkably, this marked the initial occurrence of a substantial MVD outbreak and the primary instance of a combined outbreak involving Marburg virus and Ravn virus. These two closely related viruses both contribute to causing MVD .3.6.Rousettus aegyptiacus, and the first definite filovirus reservoir was discovered by sampling bats . An epidemic that occurred in Uganda in 2007 only resulted in four confirmed cases. The patients were Ibanda district workers at the Kitaka mine. The two workers contracted the disease after sharing a tent camp with the index case in the Kashoya-Kitomi Central Forest Reserve close to the mine. The fourth patient was working at the mine when the epidemic started, without any personal protective equipment (PPE). The mining adit was surrounded by bats, and the only personal protective equipment (PPE) accessible was a pair of gloves; masks, respirators, or goggles were not. The main source of infection was direct contact with bats or bat excretions. During this epidemic, MV was isolated from ing bats .3.7.Rousettus aegyptiacus bats were once more connected to the outbreak in 2012. It is interesting to note that the epidemic hit the Rousettus aegyptiacus bat population during the second half of the yearly viral cycle  in Gabon. However, real-time polymerase chain reaction (RT-PCR) testing at CIRMF yielded negative results for the presence of Marburg and Ebola viruses . On 12 F3.10.The socioeconomic impact of MVD outbreaks can significantly affect countries with weaker economies, leading to inadequate containment and management . Epidemi4.Rousettus aegyptiacus is the most important natural source of MV infection. In addition, some Chiroptera and Hipposideros caffer may act as infectious agents. There are several ways in which MV strains can be transmitted from bat to bat. Recent research found MV in rectal, oral, and urine samples from infected bats as well as in blood and oral samples from bats that had come into contact with humans. According to this research, MV is horizontally transmitted from infected bats to bat contacts  has an incubation period of two to 21 days. According to the World Health Organization (WHO), high fever, severe malaise, and headache are some of the sudden symptoms associated with Marburg virus infection. Patients frequently complain of muscle pain and pains at the same time. Patients may experience severe watery diarrhea, cramping in the stomach, nausea, and vomiting on the third day. Up to a week may pass during the phase of diarrhea. Patients frequently exhibit a characteristic appearance during this stage that is characterized by sunken eyes, indifferent facial expressions, and extreme sluggishness. Within 2 to 7 days of the onset of symptoms, a non-itchy rash commonly develops . Hemorrh5.1.Flu-like symptoms such a high fever (>40°C), a strong headache, chills, myalgias, and malaise appear during the generalization phase. The fifth day from the start of the disease is when this phase may last, after which there is a rapid attenuation. There have been reports of exhaustion, general malaise, appetite loss, nausea, vomiting, abdominal pain, and excessively watery diarrhea . Pharyng5.2.A prolonged high temperature and other nonspecific symptoms define the early organ phase, which lasts 5 to 13 days following the onset of symptoms. Patients may also experience conjunctival infections, edoema, tiredness, dyspnea, viral exanthems, and aberrant vascular permeability . Neurolo5.3.There are two distinct outcomes in the late stages of MV infection: either the patient enters a protracted period of recovery or the sickness is fatal. Eight to 16 days after the onset of the initial symptoms, death frequently occurs. Typically, multiorgan failure and shock are the two main causes of demise . The lat6.MVD models, and lab animals can all be used to study host pathophysiology and immune responses, as is widely recognized. Currently, NHPs , hamsters, guinea pigs, and mice have been used to establish four MV disease models. NHPs are the “gold standard” among these models because to their great susceptibility to MV infections, which are almost always fatal, and their specific clinical traits that are like those of human infections. Furthermore, it has been demonstrated , that NH6.1.Multiple human organs may be harmed by the Marburg virus disease (MVD), which can result in organ failure and death . The liv6.2.Due to the fact that the symptoms and signs of MVD overlap with those of endemic illnesses like Ebola and Lassa fever as well as infectious diseases like malaria, dengue fever, and typhoid fever, it can be difficult to diagnose. The fact that MVD symptoms cannot be present at first presentation and that fever may even go away entirely during the disease adds to its complexity . This ma7.To prevent the disease’s spread, the WHO has prescribed several measures to manage the virus . When ca7.1.The World Health Organization (WHO) advocates the implementation of vigilant surveillance for Marburg virus disease (MVD) as a preventive measure against outbreaks, alongside aiding countries at risk in formulating preparedness plans . The subHealthcare practitioners are advised to assess individuals exhibiting illness for the presence of Marburg virus disease, especially if they have been in close proximity to a person with suspected or confirmed MVD within the preceding 21 days or have visited an area with an ongoing Marburg outbreak during the same period .Healthcare providers should exercise heightened vigilance and conduct evaluations for patients displaying symptoms suggestive of MVD .Health care personnel tending to patients with presumed or confirmed Marburg virus infections are to implement supplementary infection control measures to avert contact with the patient’s blood, bodily fluids, and contaminated surfaces or items such as garments and bedding .A variety of precautions can be taken by those who live in or travel to areas where the Marburg virus may be present to protect themselves and prevent the virus from spreading. These precautions include avoiding contact with the bodily fluids of those who are ill, delaying interaction with the semen of those who have recovered from MVD until the virus has been proven to be absent, and avoiding animals that may be carriers .Marburg virus infection is categorized as a notifiable condition, necessitating the implementation of rigorous isolation precautions in instances of suspected infection. It is imperative to emphasize that Marburg virus disease is classified as a notifiable condition, mandating healthcare professionals to promptly report any confirmed case or suspected instances .7.2.The Marburg virus infection can be addressed by departmental cooperation and public awareness efforts. Public education efforts can increase understanding of the disease’s severity and methods of prevention, influencing people to adopt healthy habits and seek the right medical care . To effe7.3.Several strategies are crucial in controlling Marburg virus (MV) outbreaks, as evidenced by various scientific studies and expert recommendations. Case isolation has been identified as a potent measure; a study published by the National Center for Biotechnology Information (NCBI) emphasizes that timely case isolation can effectively contain a MV outbreak . Success8.The illness is caused by a virus that has no known cure and can only be passed from person to person or animal to animal through direct contact. Getting vaccinated is the greatest way to prevent the Marburg virus from spreading. Unfortunately, there is no Marburg virus vaccine available for use in the prevention of infection currently. Despite this, researchers have been working hard to create a vaccine for the Marburg virus, and several potential strategies have been assessed. Vaccination strategies, including the use of inactivated or destroyed Marburg virus particles, have been studied and assessed . This ap8.1.The two-day-old vaccine that prevents illness is supported by the National Institute of Health. In this investigation, a deadly dose of the Marburg virus vaccine was given to rhesus macaques. After 48 h, just two of the five to six monkeys that had received the vaccine were still alive . These i9.in vitro and using macaque models, including the Ebola virus, Lassa virus, and Marburg virus  and remdesivir, have exhibited efficacy against Marburg virus disease (MVD) through preclinical investigations. These agents function as RNA polymerase inhibitors, impeding viral replication and transcription processes . T-705 . A researg virus . This rerg virus . T-705, nfection . Mice thnfection . Vero E6nfection . A broadnfection . Respiranfection . Pyrexianfection . A combinfection . The comnfection . High tenfection ,c. Marbunfection . In summ10.Clinical trials have been conducted for some drugs to treat MV; some of these trials have been completed and others are ongoing. MV drug development has progressed much more slowly than preclinical and clinical trials for the treatment of EBOV. Antiviral effectiveness against MV infections in nonhuman primates (NHPs) has long been regarded as the gold standard to assess prospective efficacy in humans and support subsequent clinical trials, much like EBOV. Following exposure in NHPs with advanced disease, several promising strategies, such as pan-filoviral small-molecule antivirals and MV-specific monoclonal antibodies, have shown excellent success. 11.The emergence of Marburg virus (MV) has presented a complex global challenge characterized by fatal outbreaks and significant clinical hurdles. The extensive analysis conducted in this study sheds light on the multifaceted nature of MV, encompassing its origin, transmission dynamics, clinical manifestations, and the intricate interplay between the virus and its human hosts. The documented outbreaks underscore the urgency of understanding and effectively addressing the threat posed by MV to global health security. Throughout this investigation, it has become evident that the clinical challenges posed by MV are substantial, with its virulent nature leading to high mortality rates and a range of severe symptoms. The lack of specific antiviral therapies or vaccines further amplifies the urgency of comprehensive prevention and control strategies. The cases studied emphasize the need for rapid, coordinated responses involving healthcare providers, researchers, and governmental bodies to curtail the impact of MV outbreaks. The global perspective on MV’s emergence emphasizes the need for proactive, multidisciplinary approaches to prevent and mitigate its devastating impact. The lessons learned from past outbreaks underscore the importance of preparedness, collaboration, and innovation in addressing this ongoing threat to public health. Through these concerted efforts, we can hope to avert future MV outbreaks and minimize their toll on human lives.12.Looking ahead, several crucial research and intervention directions emerge from our analysis. First, continued efforts are imperative to elucidate the intricate mechanisms of MV transmission from its reservoir hosts to humans, allowing for the identification of potential intervention points. Collaborative studies integrating virology, epidemiology, and ecology will be pivotal in achieving a comprehensive understanding of the virus’s life cycle. Second, the development of effective preventative measures remains paramount. Advances in vaccine technologies offer promise for the creation of MV-specific vaccines, analogous to strategies employed against related pathogens. In parallel, research should focus on identifying small molecule antiviral compounds that can impede MV replication and spread. Third, bolstering healthcare infrastructure in regions susceptible to MV outbreaks is essential. Strengthening diagnostic capabilities, training healthcare workers in effective infection control measures, and establishing rapid response protocols are vital components of managing MV cases and preventing wider disseminations.SSr, DS, SK, and AS: concept and original draft. RR, AA, SA, SR, SSa, ZA-q, PB, AM, AR-M, and RS: reviewing and editing. RS: supervision. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. 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Mpox  ravaged the non-endemic world in 2022 with dermatological and systemic manifestations. The rapid propagation of this virus shed light on the scarcity of information for a virus that was first reported in 1958. We present the first probable neonatal case of mpox with ocular involvement. Ophthalmologists may be the first to diagnose mpox or be a part of the multidisciplinary team required for adequate work-up and treatment to prevent life-long sequelae in the neonatal population. Previously, variations of the monkeypox virus resulted in a relatively innocuous infection that was endemic to Africa. However, the latest mpox virus has taken the world by storm in the past year due to its rapid and diverse appearance without geographic confinements. The first reported case of monkeypox was in Copenhagen in 1958, in a group of monkeys originating from Africa . The firMpox is an enveloped, double-stranded DNA virus in the genus Orthopoxviridae. This genus includes the deadly variola virus , which caused over 300 million deaths worldwide. Additionally, it also includes the vaccinia virus, cowpox virus, camelpox virus, and ectromelia virus (mousepox). The mpox virus is divided into Clades; Clade I (formerly Congo Basin Clade) and Clade II (formerly West African Clade). Clade II is further subdivided into Clade IIa and Clade IIb. The current international outbreak is of Clade IIb .As the virus was previously localized to Africa, there is a lack of discourse and research regarding the manifestations, outcomes, and effective treatment for monkeypox infection. With the international involvement of mpox virus Clade IIb, resources are now being utilized to identify the differences in the present outbreak compared to those of the past. Collective sharing of cases is now the crux to building the knowledge required to optimally treat this disease process and prevent resultant permanent sequelae. We report the first suspected neonatal mpox case with ocular involvement.A full-term, 10-day-old African American female, with an uncomplicated delivery and pre-natal care, presented to the emergency department (ED) due to a new-onset progressive rash and a subjective fever. The medical, ocular, family, and social history were unremarkable, except for a history of herpes simplex virus (HSV) in the patient’s mother. She was prophylactically treated with valganciclovir 10 days before delivery. Incidentally, on the day of delivery, the patient’s mother noticed a macular lesion, which progressed to a pustular lesion on her left thumb two days after delivery Figure . The motIn the pediatric ED, the patient’s vital signs were all normal except for a fever of 100.7 F. The patient had leukocytosis of 17,100 WBCs/µL  without a left shift, regular oral intake, urine, and stool output. Physical examination was significant for numerous similar-appearing circular lesions between 1 mm to 2 mm in diameter that were umbilicated, pustular with erythematous bases located on the face (predominantly on the forehead and glabella), eyelids involving the margin, abdomen, perineum, and extremities Figure . The remOphthalmology was consulted after admission due to concerns about eyelid and ocular involvement. On ophthalmic examination, the patient blinked to light in both eyes with no afferent pupillary defect or elevated intraocular pressure. A portable slit lamp examination revealed a normal anterior ocular exam. A dilated fundus exam also revealed a normal retinal exam with sharp optic nerves and no retinal lesions. The only notable exam finding was umbilicated lesions involving the bilateral eyelid and lid margin that appeared nearly identical to the lesions on the body Figure . FluoresSepsis workup returned negative for varicella zoster virus (VZV), HSV, coronavirus-19, influenza virus, and respiratory syncytial virus. A lumbar puncture with gram stain and culture of the cerebrospinal fluid was negative for any growth. The encephalitis and meningitis panels, blood, and urine cultures were also negative. Mpox polymerase chain reaction (PCR) for the patient was inconclusive, but results from the mother returned positive. Based on the disease presentation, exposure, and inconclusive mpox PCR from the neonate, the Centers for Disease Control (CDC) was consulted for management recommendations. As the mother had a positive mpox PCR with similar lesions as the neonate, the CDC recommended initiating tecovirimat (TPOXX) treatment with lubrication of the eye with bacterial coverage for suspected neonatal monkeypox.On day 15 of life, the patient remained afebrile with crusting and healing of the lesions. She was discharged home with TPOXX, Resinol (petroleum-based lubricant), acetaminophen, moxifloxacin eye drops, and erythromycin ointment to the eyelids. The patient followed up two weeks later in an infectious disease clinic with near-resolution of the skin lesions with no evident sequelae Figure . Before the outbreak in non-endemic regions, information was scarce regarding the presentation, disease course, treatment, and outcome of the human mpox infection. According to the most recent CDC update, there have been 86,956 reported cases of mpox since January 1, 2022, of which only 1,454 cases are from endemic regions. The vast majority of non-endemic cases have been reported from the European and American  regions. The United States alone has reported 30,347 cases that resulted in 42 deaths. There is a racial predilection, with 64.07% of patients reported as Black, African American, or Hispanic persons [Globally, the World Health Organization data reveals that there is a strong predisposition to mpox among men between the ages of 18 and 44 years old (78.9% of cases). The age group of 0-4 years only makes up 0.039% of cases worldwide, with a resulting lack of information on the management of this niche population. Additionally, there are increasing reports that mpox infection may be correlated with immunosuppression . The neoMpox should be considered on the differential in a neonate presenting with a vesicular rash, in addition to the traditional differential of VZV, HSV, bacterial skin infection, and bacterial sepsis. A lumbar puncture and a full sepsis workup should be performed on a neonate less than 28 days old and presenting with fever due to the high risk of mortality from disseminated infection . Ocular The patient’s mother was the only confirmed close contact with positive mpox PCR, and she developed pustular lesions on the patient’s day two of life. Our patient’s presentation followed the expected poxvirus incubation period of 5-21 days, with the development of fever and macular lesions at day seven of life . On initCurrently, no standard treatment guideline exists for mpox-related ophthalmic disease. Limited data are available regarding the efficacy of mpox treatment with antivirals, and its use has been typically reserved for those with severe disease . TPOXX aMost cases of mpox have been self-limited and managed with supportive therapy . HoweverMpox infection in the pediatric population is rare, and the clade IIb variant continues to mutate at a rate more than six to 12-fold greater than expected . We sharMpox in newborns is rare, and maintaining a high degree of clinical suspicion for new-onset vesicular lesions is essential for preventing long-term sequelae. In addition, monitoring for the involvement of lesions in and around the eye is crucial for preventing vision loss. To our knowledge, this is the first suspected case of neonatal mpox with ocular involvement that was determined as a diagnosis of exclusion. As the mpox outbreak progresses, we hope this case will help guide management in the future.