Archives of Virology

Reticuloendotheliosis retroviruses (REV) are known to cause immunosuppressive and oncogenic disease that affects numerous avian species. REV is present worldwide and recently has been reported in South America with cases of infected commercial flocks in Argentina. We surveyed for the presence of REV in birds from a state in the northern region of Brazil using real-time PCR. We report the first cases of REV in Brazil, detected in Muscovy ducks (Cairina moschata), wild turkeys (Meleagris gallopavo), and chickens (Gallus gallus) at a relatively high prevalence rate (16,8%). Phylogenetic analysis indicated a close relationship of this strain to variants in the United States. This study provides evidence of REV in the Amazon biome and provides a baseline for future surveillance of the virus in the region and throughout Brazil.

Ranavirus disease, caused by viruses within the genus Ranavirus (family Iridoviridae), is considered a globally emerging infectious disease linked to mass mortality events in both wild and cultured ectothermic vertebrates. Surveillance work is however limited in Asia hence prevalence and the dynamics of the disease remains poorly understood. To understand disease burden and the potential biotic and abiotic drivers in southern China region, we conducted a systematic surveillance of the ranavirus across Guangxi Zhuang Autonomous region (GAR). For this, we used a multifaceted approach involving screening of amphibians and other potential reservoirs, diagnostic tests, phylogenetic analyses, prevalence estimation, co-infection assessments, and climatic niche analyses. Over one thousand individuals were sampled across 25 sampling sites. We found ninety-two individuals from 18 species of ectothermic vertebrates to be infected with ranavirus. Two lineages were responsible - Rana nigromaculata ranavirus and Tiger frog virus were identified using phylogenetic analysis based on the major capsid protein (MCP) gene fragment. We also found evidence of a co-infection with ranavirus and Bd that can be highly detrimental to host populations; possibly the first such documentation in Asia. Our niche modelling analysis suggests that precipitation and seasonality play an important role in ranavirus prevalence in Guangxi region - southwestern, southeastern, central and northeastern regions of GAR can be considered to be optimum habitats for ranaviruses. Infection rates in wild frog species have reached 100% in some areas, even in nature reserves. Our research also indicates that culture facilities and pet farms are frequently infected, serving as likely vectors for the regional and global spread of ranaviruses. The knowledge generated suggests the need for systematic surveillance, stringent biosecurity measures, and control of international animal trade to prevent further transmission and protection of biodiversity and aquaculture industries across Asia.

Avian pox disease is a highly contagious infection caused by pox virus and has serious consequences on avian species with regards to economic and conservation aspects. This viral genus named as Avipox virus (APV) that infects nearly 300 bird species and lack of enough complete genome information creates hindrance to infer this virus biology. Thus in this study, we have revealed the first complete genome of an Indian pigeon pox virus that belongs to the genus APV followed by comparative genomics analysis. The entire genome of present isolate (PPV/Pur-Od-4b/01/Ind) having 280058 bp nucleotide sequences with the GC content 29.51%. The unique feature of this complete genome revealed the presence of 270 open reading frames (ORFs) circumscribed by inverted terminal repeats (ITRs) of 4,689 bp at each end and lack of recombination events. The concatenated amino acid phylogenetic tree deciphered the present isolate closely related with Feral Pigeon pox virus derived from Africa. The molecular markers, such as microsatellites were ubiquitously distributed throughout the genome and more prevalent within the functional genes.

Psittacine beak and feather disease virus (PBFDV) is a widespread and highly pathogenic virus in parrots (Psittaciformes), threatening both captive and wild populations over the world. The disease typically presents with feather and beak abnormalities, along with possible immune system suppression. No cure or commercialized vaccine is currently available. Our understanding of the Psittacine beak and feather disease often come from infected individuals with visible symptoms. Limited knowledge exists regarding the pathology and role of asymptomatic individuals in disease transmission. Asymptomatic individuals could shed virus in their crop secretion, feces, or feathers. In this study, we investigated the temporal change in viral load in feather and fecal samples from 17 asymptomatic rosy-faced lovebirds (Agapornis roseicollis). We developed a qPCR assay for PBFDV viral load quantification in the studied lovebirds. Our results showed that most of the individuals had very low viral load, while three individuals with high viral load at the beginning of the experiment were observed to exhibit a decreasing trend in viral load in both fecal and feather samples. Surprisingly, the viral load in an individual can drop from a high level to an undetectable level within three months, which is contrary to the prevailing notion that the disease is highly lethal with few reports of complete recovery. We also showed that viral load in feathers was higher than in feces. Our study provides valuable insights into the infection dynamics of PBFDV in asymptomatic individuals and contribute to the understanding of disease transmission in parrots.

The genetic diversity and evolutionary origin of the Monkeypox virus (MPXV) that is currently creating a multi-country outbreak-2022 is not fully understood. Here we report that the MPXVs that cause outbreak-2022 (MPXVs-2022) have deletion/insertion of [~]500 to 2000bp nucleotide in multiple genomic regions. Our analyses revealed that MPXVs-2022 are very close to the West African Clade of MPXVs (WA-MPXVs) that caused the Outbreak in Nigeria in 2017-2018. Furthermore, we classified the WA-MPXVs detected before 2017 that could not be transmitted from human-to-human as WA-MPXVs-I and WA-MPXVs detected after 2017 that could be transmitted from human-to-human as WA-MPXVs-II (including MPXVs-2022), and human-to-human transmissible Central African MPXVs (CA-MPXV) remained as a separate clade. Overall our results suggest that although WA-MPXVs-II are almost identical to WA-MPXVs-I throughout the genome and two large genomic insertions ([~]500, 2000bp size insertion), they differ from WA-MPXVs-I in 5-inverted terminal repeat (5-ITR) (deletion of the last-2000bp-5-ITR) and 13 proteins that of CA-MPXVs, and the presence of seven unique proteins in WA-MPXVs-II is likely to be a significant cause of outbreak-2022. This study shed light on the genetic diversity and evolutionary origin of MPXVs causing outbreak-2022.

The Citrus yellow vein clearing virus (CYVCV) causes a viral disease that has been reported in specific citrus-growing regions in Euro-Asia including countries of Pakistan, India, Turkiye, Iran, China and south Korea. Recently, CYVCV was detected in a localized urban area in a town in heart of Californias citrus-growing region and marks the first occurrence of the virus in North America. CYVCV is spread by aphid and whitefly vectors and is graft and mechanically transmitted. Hence, it is an invasive disease that presents a significant threat to the California citrus industry, especially lemons which are highly susceptible to CYVCV. To elucidate the origin of the CYVCV California strain, we used long-read sequencing technology and obtained the complete genomes of three California CYVCV isolates, CA1, CA2, and CA3. The sequences of these isolates exhibited intergenomic similarities ranging from 95.4% to 97.4% to 54 publicly available CYVCV genome sequences which indicated a relatively low level of heterogeneity. However, CYVCV CA isolates formed a distinct clade from the other isolates when aligned against other CYVCV genomes and coat protein gene sequences. Based on a rooted Maximum Likelihood phylogenetic tree, CYVCV CA isolates shared the most recent common ancestor with isolates from India. Further examination of 79 coat protein gene sequences collected over a 31-year period that spanned regions from East and South Asia to the Middle East and California, Bayesian evolutionary inferences resulted in a spatiotemporal reconstruction that placed the origin of all CYVCV to the 1930s, with South Asia as the most plausible geographic source. This analysis also suggested that CYVCV CA isolates diverged from Indian lineages, possibly around the 2010s. Moreover, the spatiotemporal phylogenetic analysis indicated two additional virus diffusion pathways: one from South Asia to East Asia and another from South Asia to the Middle East. Collectively, our phylogenetic inferences offer insights into the probable dynamics of global CYVCV dissemination, emphasizing the need for citrus industries and regulatory agencies to closely monitor citrus commodities crossing state and international borders. Author SummaryA localized outbreak of CYVCV was detected in a central California town, marking its first appearance in North America. The study sequenced the complete genomes of three CYVCV isolates from California and employed statistical algorithms to investigate the population dynamics and origin of CYVCV. Upon comparing coat protein gene sequences, the CYVCV isolates from California formed a distinct group separate from those found in other geological regions. The studys spatiotemporal phylogenetic analysis highlighted that CYVCV likely originated in the 1930s, with South Asia as the most plausible source. Notably, the CYVCV isolates from California diverged from Indian lineages, possibly around the 2010s. This study contributes to a better understanding of CYVCVs genetic and molecular diversity, shedding light on virus ecology, evolution, and biology.

The West Nile virus (WNV), primarily transmitted by mosquitoes, is one of the most widespread flaviviruses globally, with past outbreaks occurring in the USA and Europe. Recent studies in parts of Africa, including Kenya, have identified the West Nile virus Koutango lineage (WN-KOUTV) among phlebotomine sandfly populations, however, our understanding of this virus remains limited. Hence, this study aimed to characterize WN-KOUTV from phlebotomine sandflies. Sandflies were sampled between 12-16th March 2021 from six villages in Baringo South, Kenya, using CDC light traps. Female sandflies were taxonomically identified and pooled based on genus. Virus isolation was performed in Vero cells. Viral genome was determined using next-generation sequencing. Phylogenetic and molecular clock analyses were done to decipher the viruss evolutionary relationships. Comparative analyses of amino acid sequences were performed to determine variations. Protein modeling in Pymol was conducted to elucidate variations in key protein regions. Evolutionary pressure analysis investigated the selection pressures on the virus. In vitro experiments were done to investigate the virus growth kinetics in mammalian (Vero-E6) and mosquito (C636) cells. We report the isolation of WN-KOUTV from Salabani Baringo South, Kenya. The isolated WN-KOUTV clustered with previously identified WN-KOUTV strains. Comparative analysis revealed unique amino acid at NS5 653. Diversifying pressure was acting NS3 267 of the WN-KOUTV lineage. WN-KOUTV replicates efficiently in Vero-E6 and C636 cells comparable to West Nile virus Lineage 1a, isolated from mosquitoes. The isolation of WN-KOUTV in sandflies points to them as potential vectors, however, vector competence studies would confirm this. The efficient replication in mammalian and mosquito cell lines elucidated its adaptability to host and vector. We speculate the close genetic relationship of WN-KOUTV strains is enabled by the bird migratory route between East and West Africa. If proven, this may point to a potential future pandemic pathway for this virus.

The Totiviridae family of viruses has a unique genome consisting of double-stranded RNA with two open reading frames that encode the capsid protein (Cap) and the RNA-dependent RNA polymerase (RdRp). Most virions in this family are isometric in shape, approximately 40 nm in diameter, and lack envelope. There are five genera within this family, including Totivirus, Victorivirus, Giardiavirus, Leishmaniavirus, and Trichomonasvirus. While Totivirus and Victorivirus primarily infect fungi, Giardiavirus, Leishmaniavirus, and Trichomonasvirus infect diverse hosts, including protists, insects, and vertebrates. Recently, new totivirus-like species have been discovered in fish and plant hosts, and through metagenomic analysis, a novel totivirus-like virus (named Tianjin totivirus) has been isolated from bat guano. Interestingly, Tianjin totivirus causes cytopathic effects in insect cells but cannot grow in mammalian cells, suggesting that it infects insects consumed by insectivorous bats. In this study, we used next-generation sequencing and identified totivirus-like viruses in liver tissue from Molossus molossus bats () in the Amazon region of Brazil. Comparative phylogenetic analysis based on the RNA-dependent RNA polymerase region revealed that the viruses identified in Molossus bats belong to two distinct phylogenetic clades, possibly comprising different genera within the Totiviridae family. Notably, the mean similarity between Tianjin totivirus and the totiviruses identified in Molossus bats is less than 18%. These findings suggest that the diversity of totiviruses in bats is more extensive than previously recognized and highlight the potential for bats to serve as reservoirs for novel toti-like viruses.

Borealpox virus (BRPV; formerly Alaskapox) is an orthopoxvirus that has caused seven reported human infections in Alaska since 2015, including a fatal case in 2023. The natural reservoir of BRPV is unknown, although previous investigations have raised the possibility of wild small mammals transmitting the virus to humans, either through direct contact or via domestic cats and dogs. To understand which species may be involved in the maintenance and/or spillover of BRPV in Alaska, we trapped and sampled wild small mammals (including voles, shrews, and squirrels) in 2021 and 2024 near reported human case locations in Fairbanks and the Kenai Peninsula, respectively. We found evidence of previous exposure to orthopoxviruses in five species (including the House Mouse, Mus musculus) and detected BRPV DNA as well as viable virus in Northern Red-backed Voles (Clethrionomys rutilus). Further, screening of tissues from historical museum specimens revealed BRPV DNA in C. rutilus specimens collected in Denali National Park and Preserve in 1998 and 1999, 17 years before the first reported human case of BRPV. Phylogenomic analysis of all human and animal BRPV isolates strongly supports the hypothesis of local human infections through multiple spillover events. These findings suggest C. rutilus as a possible reservoir species for BRPV and indicate that BRPV has been present in Alaskan wild small-mammal populations for at least 25 years. Our study highlights the potential of museum collections to elucidate the temporal, spatial, and host ranges of emerging pathogens. Further museum- and field-based sampling will clarify the true geographic range of BRPV, which is closely related to Old World orthopoxviruses and may be circulating beyond North America.

The ongoing worldwide monkeypox outbreak is caused by viral lineages (globally referred to as hMPXV1) that are related to but distinct from clade IIb MPXV viruses transmitted in Nigeria. Analysis of genetic differences indicated that APOBEC-mediated editing might be responsible for the unexpectedly high number of mutations observed in hMPXV1 genomes. Here, using 1624 hMPXV1 publicly available sequences, we analyzed mutations that accrued since 2017 until the emergence of the current predominant variant (B.1), as well as those that that have been accumulating during the 2022 outbreak. We found that substitutions tend to cluster and mutational hot-spots are observed. Investigation of the sequence context of C to T changes indicated a preference for 5-TCA/G-3 motifs, suggesting APOBEC3F- or APOBEC3A-mediated editing. The sequence context has remained unchanged since 2017, indicating that the same mutational mechanism that is driving the accumulation of substitutions during the ongoing human-to-human transmission, was already operating before the virus left Africa. We suggest that APOBEC3A is the most likely candidate, given its expression in the skin and its known role in the editing of human papillomavirus.

Reptarenaviruses are viruses belonging to the genus Reptarenavirus within the family Arenaviridae, which infect snakes and cause inclusion body disease (IBD), a fatal condition characterized by behavioral abnormalities and wasting. Although many reptarenaviruses have been identified thus far, the phylogenetic gaps between reptarenaviruses and the other arenaviruses suggest the existence of yet-to-be-identified reptarenaviruses filling the gaps. In this study, we identified a novel reptarenavirus from publicly available RNA-seq data derived from Amazon coral snake (Micrurus spixii) and tentatively named it Amazon coral snake virus 1 (ACSV-1). We identified four ACSV-1 contigs containing the putative full-length open reading frames of the NP, GP, and L genes, as well as the partial Z gene. Phylogenetic analyses showed that ACSV-1 is highly divergent from known reptarenaviruses. The NP, GP, and L genes showed 48.3%, 42.3%, and 45.7% nucleotide sequence identities, respectively, with those of the closest relatives. Based on the International Committee on Taxonomy of Viruses (ICTV) species demarcation criteria, ACSV-1 can be assigned to a novel species of virus within the genus Reptarenavirus. This study expands our understanding of the diversity and evolution of reptarenaviruses.

Foot and mouth disease is highly contagious and notifiable transboundary disease of cattle that can cause a huge cattle productivity and production loss. A cross-sectional study was performed to estimate sero prevalence, assess associated risk factors and molecular detection of FMDV in cattle. Cluster sampling technique was employed for the selection of sampling units for the seroprevalence study. A total of 245 blood samples were collected using plain vacutainer tubes and the obtained sera were tested by 3ABC-Ab ELISA at the Animal Health institute. Twenty nine (29) epithelial tissue and vesicular fluid samples were collected purposively from outbreak cases for the molecular detection of FMDV. Kebeles and individual cattle were randomly selected, while households were designated using systematic random sampling method. An overall prevalence of 22.45% (95%, CI=17.22%-27.67%) was recorded. Multivariable logistic regression analysis indicated that herd size, age, new animal introduction into the herd and management system were the major risk factors, significantly associated with FMD sero positivity (P<0.05). The large herd size had 4-times (OR=3.97; P=0.000) more odds of FMD sero-positivity compared to the small herd sizes. The FMD seropositivity decreases 0.11013 as the Cattle age increases by 5years with the (coefficient=-0.11; P=0.172). The animals from herd to which new animals was introduced had nearly 9-times more odds (OR=9.40; P=0.000) of sero positivity than the animals sampled from no new animal introduction. Likewise, cattle those reared under extensive management system were 4-times (OR=4.10; P=0.009) at higher chance of being sero-positive compared to the intensive one. From outbreak cases, 27 (93.1%) were identified positive for FMDV serotype SAT 2. A total of 124 individuals were interviewed, and the majority responded that there is no practice of reporting disease outbreak, free animal movement, free rangeland grazing and they use traditional case management as a means of controlling the disease. The finding of FMD virus antibodies in cattle from all study areas indicate endemic circulation of the virus. The implementation of regular vaccination could minimize the occurrence and further molecular characterization should be needed to identify other serotypes of FMD virus that could inform to supply an appropriate vaccine to the area.

Foot-and-mouth disease virus (FMDV) has proven its potential to propagate across local and international borders on numerous occasions, but yet details about the directionality of the spread along with the role of the different host in transmission remain unexplored. To elucidate FMDV global spread characteristics, we studied the spatiotemporal phylodynamics of serotypes O, A, Asia1, SAT1, SAT2, and SAT3, based on more than 50 years of phylogenetic and epidemiological information. Our results revealed phylogeographic patterns, dispersal rates, and the role of host species in the dispersal and maintenance of virus circulation. Contrary to previous studies, our results showed that three serotypes were monophyletic (O, A, and Asia1), while all SATs serotypes did not evidence a defined common ancestor. Root state posterior probability (RSPP) analysis suggested Belgium as the country of origin for serotype O (RSPP=0.27). India was the ancestral country for serotypes A (RSPP= 0.28), and Asia-1 (RSPP= 0.34), while Uganda appeared as the most likely origin country of all SAT serotypes (RSPP> 0.45). Furthermore, we identified the key centers of dispersal of the virus, being China, India and Uganda the most important ones. Bayes factor analysis revealed cattle as the major source of the virus for most of the serotypes (RSPP> 0.63), where the most important host-species transition route for serotypes O, A, and Asia1 was from cattle Bos taurus to swine Sus scrofa domesticus (BF>500), while, for SAT serotypes was from B. taurus to African buffalo Syncerus caffer. This study provides significant insights into the spatiotemporal dynamics of the global circulation of FMDV serotypes, by characterizing the viral routes of spread at serotype level, especially uncovering the importance of host species for each serotype in the evolution and spread of FMDV which further improve future decisions for more efficient control and eradication.

Cotton (Gossypium spp.) is a globally significant cash crop cultivated for its versatile fiber, widely used in the textile industry. Cotton, as other crops, is vulnerable to infectious pathogens. Several of them, including viruses, are a major threat to cotton production. Geminiviruses (family Geminiviridae) are insect transmitted, small non-enveloped viruses, with circular single-stranded DNA genomes, which are encapsidated in quasi-icosahedral geminated virions. Here we present evidence of a novel begomovirus (genus Begomovirus) infecting cotton from Argentina. Two circular ssDNA virus sequences were assembled from high-throughput sequencing data from Gossyipium hirsutum cotton samples showing mosaic symptoms from Argentina. Structural and functional annotation indicated that the virus sequences corresponded to complete DNA components A and B of a novel New World bipartite begomovirus. Genetic distance and evolutionary analyses support that the detected sequences correspond to a new virus, a tentative prototype member of a novel species which we propose the name "Cotton mosaic virus" (CoMV).

BackgroundIn agroecosystems, viruses are well known to influence crop health and a few cause phytosanitary and economic problems, but their diversity in non-crop plants and role outside the disease perspective is less known. An extensive virome exploration that includes both crop and diverse weed plants is therefore needed to better understand roles of viruses in agroecosystems. Such unbiased exploration is possible through viromics, which could generate biological and ecological insights from immense high-throughput sequencing (HTS) data. ResultsHere, we implemented HTS-based viromics to explore viral diversity in tomatoes and weeds in farming areas at a nation-wide scale. We detected 125 viruses, including 79 novel species, wherein 65 were found exclusively in weeds. This spanned 21 higher-level plant virus taxa dominated by Potyviridae, Rhabdoviridae, and Tombusviridae, and four non-plant virus families. We detected viruses of non-plant hosts and viroid-like sequences, and demonstrated infectivity of a novel tobamovirus in plants of Solanaceae family. Diversities of predominant tomato viruses were variable, in some cases, comparable to that of global isolates of same species. We phylogenetically classified novel viruses, and showed links between a subgroup of phylogenetically-related rhabdoviruses to their taxonomically-related host plants. Ten classified viruses detected in tomatoes were also detected in weeds, which might indicate possible role of weeds as their reservoirs, and that these viruses could be exchanged between the two compartments. ConclusionsWe showed that even in relatively well studied agroecosystems, such as tomato farms, a large part of very diverse plant viromes can still be unknown and is mostly present in understudied non-crop plants. The overlapping presence of viruses in tomatoes and weeds implicate possible presence of virus reservoir and possible exchange between the weed and crop compartments, which may influence weed management decisions. The observed variability and widespread presence of predominant tomato viruses and the infectivity of a novel tobamovirus in solanaceous plants, provided foundation for further investigation of virus disease dynamics and their effect on tomato health. The extensive insights we generated from such in-depth agroecosystem virome exploration will be valuable in anticipating possible emergences of plant virus diseases, and would serve as baseline for further post-discovery characterization studies.

Mesta yellow vein mosaic disease (MYVMD), one of the major diseases circulating mesta growing regions of Indian sub-continent, is responsible for serious yield loss in mesta crops. A complex of monopartite begomovirus, Mesta yellow vein mosaic virus (MYVMV) and associated betasatellite, is reported in several studies as the causal agent of MYVMD. However, all-inclusive molecular evolutionary analysis of so far available MYVMVs and associated betasatellites disseminating in this region is still lacking. In this study, by estimating and analyzing various indexes of population genetics and evolutionary parameters, we discussed the sources of genetic variations, population dynamics and different forces acting on the evolution of MYVMVs and associated betasatellites. The study finds recombination as a vital force in the evolution and diversification of begomovirus complexes in different geographic locations however, betasatellites were found to be exposed to more diverse recombination events compared to MYVMVs. Indian isolates are reported to have high frequency of polymorphism in this study which suggests a balancing selection or expansion occurring in Indian populations of begomoviruses. Higher degree of genetic differentiation and lower rate of gene flow calculated between the viral populations of Bangladesh and Pakistan is justified by the relatively far geographical distance between these two countries. Although the study detects overall purifying selection, the degrees of constraints acting on individual gene tested are found different. Coat protein (AV1) is estimated with very high nucleotide substitution rate which is very likely to result from the strongest purifying selection pressure (dN/dS = 0.131) calculated in this study on coat protein. The findings of this study on different evolutionary forces that shape the emergence and diversification of MYVMVs and associated betasatellites may provide directions towards future evolutionary trend analysis and development of comprehensive disease control strategies for begomoviruses.

This study explores the genomic diversity of three novel Konjac mosaic virus (KoMV) variants, originating from distinct Zantedeschia (calla lily) commercial cultivars. Virus-derived small RNA sequencing was performed and the complete KoMV variant genome sequences were determined by de novo assembly of short reads. All KoMV variants showed substantial nucleotide, as well as protein differences as compared to the KoMV RefSeq sequence.

BackgroundOn May 7, 2022, a case of monkeypox virus (MPXV) has been reported to the WHO. It causes a viral zoonotic disease with characteristics comparable to that of smallpox cases. Monkeypox could be a serious health concern as it is already spreading to multiple countries worldwide. To prevent the monkeypox infection and for its early detection, a potential vaccine and diagnostic candidate is urgently required. MethodIn present study we have used different in silico methods to screen the entire genome with its approximately 191 genes for potential epitopes present in proliferation and virulent proteins of monkeypox virus. All protein sequences were retrieved from different genomic and proteome databases listed in Uniprot or NCBI. ResultsIn the present study we have screened potential epitopes from 11 different proteins of Monkeypox. All the included protein play an important role in pathogenesis and/or proliferation of Monkeypox virus. We have identified in total 984 CTL and 168 HTL epitopes with highest score in our epitope screening. The reported epitopes could be potential candidates for the design of an early detection diagnostic kit specific for the monkeypox virus. Out of these target peptides we have included a total of 39 CTL epitopes and 39 HTL epitopes in design of multi-epitope vaccine candidates. These shortlisted epitopes are highly conserved amongst different strains and origin of monkeypox viruses. The population coverage by joint administration of CTL and HTL MEVs is predicted to be high with the epitopes showing potential to bind upto 24 different CTL and HTL HLA allele molecules. The epitopes used in MEVs are examined to be highly immunogenic, non allergic but antigenic, and non toxic. All the CTL and HTL MEVs designed utilizing the epitopes have physiochemical properties favor its over expression in human cells. The optimized cDNA constructs of CTL and HTL MEVs also favor over expression of MEVs in human cells. Overall, the MEV construct proposed by us are fissile for expression in the lab and for further in vivo studies. ConclusionControl and fight against emerging diseases such as MPXV requires pathogen diagnostic and novel vaccine approaches. We screened for several epitopes and designed a MEV providing a potential solution for both purposes. Our method allows rapid screening and provides a rational strategy for the development of vaccine candidate effective in fighting MPXV and other unexpected upcoming diseases.

Lloviu cuevavirus (LLOV) was the first identified member of Filoviridae family outside the Ebola and Marburgvirus genera. A massive die-off of Schreibers bent-winged bats (Miniopterus schreibersii) in the Iberian Peninsula in 2002 led to its discovery. Studies with recombinant and wild-type LLOV isolates confirmed the susceptibility of human-derived cell lines and primary human macrophages to LLOV infection in vitro. Based on these data, LLOV is now considered as a potential zoonotic virus with unknown pathogenicity to humans and bats. We examined bat samples from Italy for the presence of LLOV in an area outside of the currently known distribution range of the virus. We detected one positive sample from 2020, sequenced the complete coding sequence of the viral genome and established an infectious isolate of the virus. In addition, we performed the first comprehensive evolutionary analysis of the virus, using the Spanish, Hungarian and the Italian sequences. The most important achievement of this article is the establishment of an additional infectious LLOV isolate from a bat sample using the SuBK12-08 cells, demonstrating that this cell line is highly susceptible to LLOV infection. These results further confirms the role of these bats as the host of this virus, possibly throughout their entire geographic range. This is an important result to further understand the role of bats as the natural hosts for zoonotic filoviruses.

Kindia tick virus (KITV) is a novel multicomponent virus first detected by direct sequencing of Rhipicephalus geigyi ticks in Guinea in 2017. Here, we present a complete coding genome sequence for all four segments of KITV/2017/1. This virus appears to be evolutionarily related to unclassified flaviviruses, such as Alongshan virus.