Contamination of oysters with a variety of viruses is one key pathway to trigger outbreaks of massive oyster mortality as well as human illnesses, including gastroenteritis and hepatitis. Much effort has gone into examining the fate of viruses in contaminated oysters, yet the current state of knowledge of nonlinear virus-oyster interactions is not comprehensive because most studies have focused on a limited number of processes under a narrow range of experimental conditions. A framework is needed for describing the complex nonlinear virus-oyster interactions. Here, we introduce a mathematical model that includes key processes for viral dynamics in oysters, such as oyster filtration, viral replication, the antiviral immune response, apoptosis, autophagy, and selective accumulation. We evaluate the model performance for two groups of viruses, those that replicate in oysters  and those that do not , and show that this model simulates well the viral dynamics in oysters for both groups. The model analytically explains experimental findings and predicts how changes in different physiological processes and environmental conditions nonlinearly affect in-host viral dynamics, for example, that oysters at higher temperatures may be more resistant to infection by ostreid herpesvirus. It also provides new insight into food treatment for controlling outbreaks, for example, that depuration for reducing norovirus levels is more effective in environments where oyster filtration rates are higher. This study provides the foundation of a modeling framework to guide future experiments and numerical modeling for better prediction and management of outbreaks.IMPORTANCE The fate of viruses in contaminated oysters has received a significant amount of attention in the fields of oyster aquaculture, food quality control, and public health. However, intensive studies through laboratory experiments and in situ observations are often conducted under a narrow range of experimental conditions and for a specific purpose in their respective fields. Given the complex interactions of various processes and nonlinear viral responses to changes in physiological and environmental conditions, a theoretical framework fully describing the viral dynamics in oysters is warranted to guide future studies from a top-down design. Here, we developed a process-based, in-host modeling framework that builds a bridge for better communications between different disciplines studying virus-oyster interactions. The recognition of the importance of virus-oyster interactions is increasing, and the dynamics of viruses in oysters after infection have been recognized as a key linkage to the massive mortality of oysters  as well A group of viruses (referred to as “group 1” here) that can infect oyster cells and replicate in oysters, such as ostreid herpesvirus 1 (OsHV-1), can have harmful impacts on oysters and, therefore, on oyster aquaculture industries. These viruses, particularly the OsHV-1 μvars, have caused massive mortalities of farmed Pacific oysters and severe economic losses in Europe, New Zealand, and Australia , 4. The 5–Other viruses do not replicate inside the shellfish and do not have adverse effects on the host (referred to as “group 2”). These include viruses such as norovirus (NoV), Tulane virus (TV), hepatitis A virus, and feline calicivirus. However, some viruses in this group, like human enteric viruses , have been found to potentially impact human health through the consumption of shellfish like oysters and, therefore, threaten the seafood industry . This isThere is evidence that oysters are not passive filters or ionic traps but rather that they have specific ligands that selectively accumulate viruses . Therefo18–Thorough knowledge of the interaction between viruses and oysters is needed to support management decisions , which hAlthough the importance of marine viruses in the ecosystem is recognized, there are few mathematical models describing oyster-virus interactions within oysters. Many existing models, including those examining virus-host interactions in marine systems, are focused on viral transmission between host individuals and host population dynamics, and most of them are developed for studying the population dynamics of organisms other than shellfish, such as phytoplankton and bacterioplankton . In thesHere, we propose a new modeling framework that includes all essential processes when oysters are contaminated by viruses. The model is for in-host viral dynamics in one oyster and investigates virus infection between targeted tissue cells in that oyster.T (grams per oyster); (ii) the infected cell, I (grams per oyster); and (iii) the virus concentration in oyster tissues, V (copies per gram), respectively:T0, I0, V0). The definition of each parameter is listed in t (days) denotes time. m (grams per oyster) is the weight of the total target cells in one oyster if the oyster is not infected. β (grams per copy per day) is the infection rate. Note that β is considered on the scale of host cells within an oyster, which reflects the rate of entry of the virus into the host cells, a concept that differs from the concept of virus infectivity or virulence in studies focusing on virus transmission between host individuals. δT (per day) and δ (per day) are the death rates of uninfected and infected target cells, respectively, and it is naturally assumed that for oyster-pathogenic viruses, δ is less than or equal to δT. Ts (grams per oyster per day) is the production rate of new uninfected target cells. New uninfected cells can be generated (i) by the replacement of old cells that have been eliminated through natural death (δTT), (ii) by increasing cell numbers through oyster growth, and (iii) as a host response to infection that leads to an increase in cells, such as hemocyte infiltration in response to pathogens or at wound sites in oysters. p (copies per gram per day) is the rate of virus replication in infected cells. Because virus replication requires the infection of target cells, and this process results in the loss of those viruses from the oyster tissues through either the absorption of the viral particle into the cell or the injection of viral components such as nucleic acids into the cell to induce the production of new virus (a/m)βTV; a (copies per gram) describes how many copies of viruses are needed to infect 1 g of target cells. lV (copies per cubic meter) is the virus concentration in the surrounding water, f (cubic meter per day per oyster) is the filtration rate, and c (per day) is the in-host virus clearance/elimination rate. σ (cubic meters per oyster) is the volume of the individual oyster. εi is the bioaccumulation fraction in the filter-in process; εo is the fraction of virus that cannot be retained in oyster tissues during filter-out/shedding processes, referred to as the shedding fraction; and the two parameters depend on both selective accumulation and in-host physical trapping. An εi value of 1 means that all particles that are filtered into the oyster bind the tissues, while an εi value of 0 means that no filtered-in virus can accumulate in oyster tissues. An εo value of 0 means that the process of filtration cannot remove virus particles from the oyster body, while an εo value of 1 means that virus particles in oyster tissues are freely filtered out. Clearly, εi and εo show a negative relationship; i.e., a higher εi corresponds to a lower εo. In pI/m) − (a/m)βTV describes the in-host production of copies of the virus by infected cells, and the terms (εi/m)lfV − (εo/σ)fV and −cV describe net virus uptake through filtration  and in-host clearance, respectively.In-host dynamics are governed by three coupled equations, which are (i) the uninfected target cell, ew virus , a loss p = 0 so that lV is the variable connecting to the aquatic system, and it can be linked to a dynamic virus transport model in the water.For group 2 viruses , β = V that readsq = (p/m/δ − a/m)(Ts − δTT*), which represents the steady-state net in-host viral production, in addition to the filter-in, (εi/m)lfV, from the water. All parameters in lV is zero, there is another trivial equilibrium (V* = 0), indicating that the virus can be removed completely from the oyster tissues. Whether V approaches trivial or nontrivial equilibrium depends on the particular conditions . For group 2 viruses, q is zero, and V* is proportional to lV. If lV becomes zero, V* approaches zero.In natural waters, oysters are usually cultured for a relatively long time, and it is sometimes necessary to study the long-term accumulation of viruses in oysters. With long-term exposure to a virus, the concentration in oysters is expected to be varying around equilibrium values. Long-term viral dynamics for either group 1 or 2 viruses can be examined by the nontrivial steady-state solution for Many abiotic and biotic factors impact the various processes regulating viral accumulation and dynamics in oysters, including virus type; oyster factors such as size, age, species, the selective accumulation of some viruses, and host antiviral responses; and environmental factors such as temperature and salinity. These factors and their impacts are accounted for in the model through changes in parameter values.lV, is determined by the fate and transport of the virus following discharge into the system. This means that the viral concentration in oyster tissues is determined by the overall concentration of the virus in the water rather than the loading pattern , which agrees with previously reported experimental results  if observational data are available. The function can vary among oyster species  and an increase in clearance .Viruses use host metabolic machinery for replication, and their replication rate, er cells . p is ale system , and thee system . The oyse system  and apope system . These pp to δ in q in It has been suggested that apoptosis is an important defense mechanism that the oyster uses against OsHV-1 by limiting viral spread and eliminating infected cells . InhibitC. gigas genome are responsive to changes in environmental conditions  depends on the virus type, oyster species, and mechanisms of entry of the virus into host cells. The envelopes of some virus types can promote transmission from one host cell to another . In addii, and the shedding fraction, εo) varies among virus species  was estimated to be about 60-fold higher than the εi for TV (ranging from 3.23 × 10−6 to 1.61 × 10−5).Using NoV and TV as two representatives of group 2 viruses, the model describes the depuration process as observed in the laboratory well . The calz et al.  , will generally shorten the time required to reach the peak and increase the peak level of the virus in oyster tissues (o/σ)f + c, will decrease V , V* always increases with f, but V* may either increase or decrease with f for group 1 viruses. Also, an increased filter-in value, (εi/m)lfV, will shorten the time to reach bioaccumulation equilibrium for group 2 viruses and contribute more to the dynamics of group 1 viruses than with the net in-host viral production denoted by q.For long-term equilibrium, p, or infection rate, β; a higher in-host elimination rate, c; and a lower production rate of new uninfected target cells, Ts, in adult oysters than in juvenile oysters. Also, the resistance of oysters to ostreid herpesvirus disease varies among different oyster species and stocks and differs with exposure to OsHV-1 strains, including among the different OsHV-1 μvars . This is demonstrated by the model parameters of the two families A1 and A2 in the case study , that is, on the order of 0.1 day−1  for C. gigas to temperature follows the measurement reported previously by Bougrier et al.  and for the infection rate (kβ) and also the in-host elimination rate (c) in the model. Particularly, it is suggested that higher temperatures may reduce the viral levels by limiting OsHV-1 entry into oyster cells (0). The death rate of infected target cells (δ) is highly controlled by apoptosis. It is regulated by both pro- and antiapoptotic effects in oysters, which in turn may be influenced by the presence of the virus itself. The expression of both pro- and antiapoptotic genes is upregulated with an increase in temperature f + c]. Without considering any changes in the virus concentration in the surrounding water (lV), the equilibrium viral level decreases with temperature in all three scenarios. It is interesting to examine how temperature impacts the maximum virus concentration in the oyster (Vmax) during the acute phase and the time to reach Vmax  is the ratio of the total target cells in the infected oyster to the total target cells if the oyster has not been infected, and a very low T′ + I′ value may result in a low chance of survival during infection. For example, studies show that a high mortality rate of virally infected shrimp occurs when the density of the total hemocytes becomes significantly decreased (T′ + I′)min, is generally positively correlated with temperature for all the three scenarios min at low temperatures in the three scenarios indicates, from a numerical modeling perspective, that apoptosis may play an important role in regulating viral proliferation in oysters. In addition, among the three scenarios, scenario 2 has the highest δ and the largest values of (T′ + I′)min at low temperatures. This indicates that those oysters having a greater net proapoptotic effect might be better able to survive at low temperatures. Note that a virally induced decrease in the number of target cells may be only one factor contributing to oyster death. For example, as proposed previously by de Lorgeril et al. (T′ + I′)min may not necessarily decrease the chances of oyster survival. This was demonstrated in a laboratory experiment where juvenile C. gigas oysters were exposed to OsHV-1, and maximum mortalities were observed at between 16°C and 22°C, with lower mortality rates above and below that temperature range  with clean flowthrough water for a relatively short period of time f + c, and hence varies with the in-host elimination rate, c; the filtration rate, f; and the shedding fraction, εo. Thus, any changes leading to a higher value of (εo/σ)f + c can result in a more rapid decrease in virus levels in oyster tissues f contributes substantially to the virus removal rate.Environmental conditions also affect these parameters and, therefore, the depuration efficacy. Within a certain range, as temperature increases, both us types , 68. Nevirginica  and 19°CC. gigas . This inC. gigas , the depf, is impacted. An increased f enhances efficacy. This may explain the results of experiments where feeding oysters with phytoplankton facilitates a decrease in virus levels in oysters compared to those without feeding lfV. Thus, both the source and sink terms during bioaccumulation are impacted by environmental conditions, which need to be studied on a case-by-case basis. Specifically, for those laboratory experiments with a high viral load for bioaccumulation, the increase of V is dominated by (εi/m)lfV, and V increases with a higher bioaccumulation fraction, εi. Also, if laboratory experiments are conducted for examining how the bioaccumulation efficiency varies with temperature, εi is expected to be fixed for particular virus and oyster species during the short experimental period, and the bioaccumulation efficiency will be positively correlated with f and show a non-monotonic relationship with temperature, as simulated by our model experiment f + c, which indicates that the estimated removal rate has low uncertainty. The viral dynamics are not sensitive to changes in s′Tnet, indicating that this parameter has high uncertainty; however, the insensitivity also suggests that the uncertainty of this parameter has little impact on the model results or its implications. Overall, uncertainties exist and vary among different parameters. The uncertainty analysis warrants further examination in future studies. As data are expanding as a result of more intensive studies, the accuracy of parameter values can be expected to be improved. In addition, studies with different species and stocks of oysters and different strains of viruses will provide additional information on the host-virus interaction. Different oyster species and stocks have various susceptibilities to different variants of oyster herpesviruses, for example  an external parameter rather than explicitly simulating it. If an explicit simulation of lV is needed for a realistic case, it is straightforward to couple the in-host model with other models that simulate lV explicitly, such as disease transmission/epidemic models  phase of infection or explicitly consider the effects of a type I IFN-like immune response on viral replication and spread , 70. NevIn this study, we developed a process-based model to provide a modeling framework for fully describing viral dynamics in oysters. The in-host model builds a bridge for better communications between different disciplines studying virus-oyster interactions and theoretically describes complex interactions of various in-host processes and nonlinear viral responses to changes in physiological and environmental conditions. The model can be used to revisit the relationships between viral dynamics and various factors by experiments, predict unknown relationships, and help to propose hypotheses and design experiments. It can also be coupled with population models and transmission models to investigate oyster population dynamics under viral infection and the transmission of viruses between oysters, which will help with better prediction and management of outbreaks. In addition, this modeling framework can be easily adapted for studying other shellfish species such as clams.A mathematical model was developed for providing a framework for studying in-host viral dynamics in oysters. For both virus groups , filtration is the process through which viruses in the water are transferred to oysters (filter-in) and viruses in oysters are moved out of the oysters to the surrounding water . It is assumed that virus particles primarily enter and leave oysters through filtration. A fraction of the filtered-in virus will be accumulated in the oyster tissues, and it is this portion that actually contributes to the viral dynamics in oysters, while the remaining portion is directly removed during filtration. Laboratory experiments suggest that this fraction is generally very small . The fraT′ (cells/oyster); infected cells, I′ (cells/oyster); and the virus concentration in oyster tissues, V̂ (virus copies/oyster), respectively:T̂0, Î0, V̂0).For group 1 viruses, the in-host viral dynamics after initial infection can be described by the widely used basic model of human virus infection , 70, witp̂ = 0. Thus, V̂:V̂0.The governing equation for the dynamics of group 2 viruses is a specific case of p in the model must also be adjusted. We denote V the virus concentration in oyster tissues in one reported unit, distinguishing it from V̂ in units of virus copies per oyster, and also T and I the uninfected and infected target cells in reported units, respectively. Next, we set θ and ω as the conversion factors to convert the units of copies per oyster and cells per oyster to the new set of units, respectively, i.e., V=θV̂, T=ωT̂, and I=ωÎ. Substituting the expressions V̂=V/θ, T̂=T/ω, and Î=I/ω into the governing p=p̂/ω, a=â/ω, and Ts=ωTŝ, T0, I0, V0) = . It can be seen that by doing this, only the conversion factor θ appears explicitly in two terms in In practice, the units of virus concentration in oysters may be studied or reported in units other than virus copies per oyster, such as virus copies per gram of tissue. In this case, the units of parameters β and ^θ)TV−δIdVdt=θ, where m (grams per oyster) is the weight of the total target cells in one oyster if the oyster is not infected and ω (grams per cell) denotes the weight of one oyster cell. β, p, α, and Ts correspondingly have the units of grams per copy per day, copies per gram per day, copies per gram, and grams per oyster per day, respectively. Specifically, if the new units are grams per oyster for The equation for group 2 viruses becomesdVdt=εimfdT/dt = dI/dt = dV/dt = 0. The steady-state solution for V readsq = (p/m/δ − a/m)(Ts − δTT*) and T* is the steady-state solution for T. The values of all other parameters in V* = 0) also exists if lV is zero.For group 1 viruses, the virus level in oysters approaches an equilibrium in surviving oysters, and it can be solved by the steady-state solution of dV/dt = 0), which readsThe long-term accumulation of group 2 viruses can be estimated by solving The stabilities of the long-term equilibriums for group 1 and group 2 viruses are analyzed in the supplemental material.T and I, which are often unreported in observations. For T and I in any given units, by setting T′ = T/aT, and I′ = I/aT, where aT is the initial number of total target cells if the oyster is not infected, having the same units as T. From aT = Ts/δT when V is zero. Using the same procedure to obtain V0), where p′=apT=p̂(aT/ω), a′=aaT=â(aT/ω), and s′ T=s′ T/aT=(ω/aT)Tŝ. Correspondingly, the long-term equilibrium of V isq′ = q in −m1.The parameters in the model can be calibrated against observational data from laboratory experiments. To better use the in-host model, it may be helpful to normalize T, is sufficiently low, and it can be neglected in the calculation by accounting for the loss implicitly in s′Tnet, i.e., Tnet can be considered the net production of uninfected target cells due only to mechanisms 2 and 3 listed in Results, which is assumed to be constant during the short period of the acute phase. The loss term (−θa′βT′V) may also be considered small enough to be neglected during the acute phase. Thus, we haveV0). For experiments starting from low V0 and For group 1 viruses, in the acute infection phase, the dynamics of the virus concentration in oysters is regulated by dominant processes. To reduce the uncertainties in calibrating parameters that do not have critical effects on the viral dynamics, we can reduce the number of parameters in the model by retaining only the dominant processes when simulating the acute phase. It is reasonable to assume that the death rate of an uninfected target cell, δp and β in the forms p0 and β0 denote the viral replication rate and the infection rate at the beginning, respectively; pk and kβ have units of per day. For the parameter θp′ in 0, pk, kβ, δ, s′Tnet, θεifVl, and (εo/σ)f + c. Note that the filter-in (θεifVl) is comparably smaller than in-host viral production, and its role becomes significant only after the acute phase; thus, for the purpose of fitting, θεifVl can be simply set constant at the long-term equilibrium value without causing large biases.For the acute infection phase in the experiments, a type I IFN-like immune response can cause decreases in viral replication and spread . WithoutVOsHV are the reported and converted OsHV-1 concentrations, respectively. The concentrations of NoV and TV reported previously by Drouaz et al.  over an experimental period can be computed with V at the end of the period and the known values of lV, f, m, and the removal rate, (εo/σ)f + c. We computed εi for NoV and TV in the DT of oysters over the first hour of the bioaccumulation experiments, where V values at 1 h are 537 and 176 viral RNA copies/g DT for NoV and TV, respectively , during the short initial period, the decrease in lV can be neglected, and lV can be assumed to be constant in the computation. As we focused on the DT, m is the mean DT weight of an individual oyster, equal to 0.53 g DT oyster−1. The removal rates, (εo/σ)f + c, were computed based on the 8-day depuration experiments following the bioaccumulation experiments under similar conditions. In fact, given the small values of the removal rates (on the order of 0.1 day−1) and the short period considered (1 h), the effect of the removal rate on the bioaccumulation of V is negligible. This is shown by the fact that i due to the uncertainty of the removal rate is limited by using f, was computed using an empirical equation for the filtration rate of C. gigas (f of C. gigas, in comparison to C. virginica, for which a low TSS can reduce its f to 0.1× f (f at least in half in the bioaccumulation experiment reported by Drouaz et al. (f values from 0.0089 to 0.044 m3 day−1 oyster−1 for the bioaccumulation experiments.We also calculated the bioaccumulation abilities of NoV and TV in the bioaccumulation experiments reported by Drouaz et al. . By assuectively . Note thC. gigas  at expero 0.1× f . Our calz et al.  by compahttps://doi.org/10.6084/m9.figshare.19294580.v1.The MATLAB code and data used during the present study are available in figshare at


Planococcus comstocki mealybug did not reproduce the strong symptoms observed on the donor plant infected with a severe form of GLD. This result raises questions about the contribution of each virus to the symptomatology of the plant.Grapevine leafroll disease (GLD) is one of the most economically damaging virus diseases in grapevine, with grapevine leafroll-associated virus 1 (GLRaV-1) and grapevine leafroll-associated virus 3 (GLRaV-3) as the main contributors. This study complements a previously published transcriptomic analysis and compared the impact of two different forms of GLD to a symptomless control treatment: a mildly symptomatic form infected with GLRaV-1 and a severe form with exceptionally early leafroll symptoms (up to six weeks before veraison) infected with GLRaV-1 and GLRaV-3. Vine physiology and fruit composition in 17-year-old Pinot noir vines were measured and a gradient of vigor, yield, and berry quality (sugar content and berry weight) was observed between treatments. Virome composition, confirmed by individual RT-PCR, was compared with biological indexing. Three divergent viromes were recovered, containing between four to seven viruses and two viroids. They included the first detection of grapevine asteroid mosaic-associated virus in Switzerland. This virus did not cause obvious symptoms on the indicators used in biological indexing. Moreover, the presence of grapevine virus B (GVB) did not cause the expected corky bark symptoms on the indicators, thus underlining the important limitations of the biological indexing. Transmission of GLRaV-3 alone or in combination with GVB by  Vitis vinifera) has been propagated and distributed throughout the world for centuries since its domestication, which is believed to have taken place about 8000 years ago [Closteroviridae. The biggest contributors to GLD are grapevine leafroll-associated virus 1 and 3 (GLRaV-1 and GLRaV-3) both belonging to the genus Ampelovirus. The name GLD can be attributed to the distinctive symptoms observed in susceptible cultivars after veraison. Additional symptoms include the early reddening of the leaf on red cultivars and a severe reduction in yield [Grapevine  has revolutionized the field of virology by providing an unprecedented tool for the detection and identification of viruses including novel viruses. However, at the same time, virology faces new bottlenecks caused by the imbalance between the large number of viral sequences available and the lack of corresponding biological data ,10.Here, to complement a transcriptomic analysis reported previously , we repoVitis berlandieri × Vitis riparia cv. Kober 5BB (referred as 5BB thereafter) and Vitis riparia × Vitis rupestris cv. 3309 Couderc (referred as 3309C thereafter). The experimental plot was grafted in 2005 and consisted of 10 replicate vines per treatment  per rootstock  for a total of 60 plants. The experiment was conducted at the Agroscope research station in Nyon, Switzerland. The different vines were planted on three adjacent rows in an area with homogeneous soil conditions. Vines were trained in the gobelet training system  with a planting density of 5500 vines per hectare (0.8 × 1.6 m). Four shoots per vine were maintained.The two virus sources used in our experiment were collected in 1984 on two single vines from Pinot noir in a vineyard. The vines showed contrasting symptoms of grapevine leafroll disease (GLD). One plant denoted GLD_S+ showed mild leafroll symptoms expression and was tested positive for the virus GLRaV-1 whereas another showed strong leafroll symptoms (GLD_S++) and was tested positive for GLRaV1, GLRaV-3, and GVB. A symptomless control was obtained by thermotherapy in order to set up a long-term experimental study to evaluate the agronomical and physiological impact of leafroll infection. The materials from the two infected vines together with symptomless control were grafted onto two certified rootstocks: −2 s−1) on symptomless leaves without virus symptoms when possible. Relative leaf chlorophyll concentrations were measured on mature leaves using a chlorophyll tester called N-tester ; the measurements (expressed as N-tester units) were taken between June and September. At commercial maturity, grapes were collected and weighed individually for each plant. Grapes were then manually pressed to extract juices, and fruit composition parameters ) were measured using Fourier transform infrared spectroscopy . Measurements of total anthocyanin content were performed according to the Glories method [Several parameters were assessed during the 6 years (2014–2019). Vine vigor and mortality were assessed on both rootstocks 3309C and 5BB. For gas exchange, fruit composition, and yield, only rootstock 5BB was evaluated. During winters, pruning wood was weighed for each individual plant. Trunk diameters were measured at ca. 10 cm above the soil level. Gas exchange measurements were conducted on two adult non-senescent leaves per plant on four plant per treatment using a LICOR 6400 XT portable photosynthesis system , as described previously . Net phos method : brieflys method . Vine mohttps://www.r-project.org/, accessed on 17 June 2022). Statistical analysis was performed by ANOVA followed by mean comparisons using Duncan test from R package Agricolae [Physiological and fruit composition data were analyzed using R software , Kober 5BB, and V. rupestris St. George. Accessions were graft-inoculated onto woody indicators. Symptoms from eight replicates were evaluated in the field over a 3-year period. Positive and negative controls used in the assay were selected from the Nyon grapevine collection [Biological indexing was performed for the two virus sources and the symptomless control following standard practices for certification . Brieflyllection .Vitis vinifera (PN40024 12X v2). Unmapped reads were de novo assembled and annotated by BlastN and BlastX analysis against reference sequences for viruses and viroids using Geneious v11.1 . In the case of GLRaV-1 and grapevine asteroid mosaic-associated virus (GAMaV), contigs were extended by additional rounds of reads mapping. Finally, in order to assess the representation of every virus/viroid in a library, the total number of reads were mapped against the assembled sequences or a similar reference genome from GenBank using the following parameters: medium low resolution and two iterations. For some genome regions of GLRaV-1 and GAMaV, the accuracy of the reconstructed genome was verified by direct Sanger sequencing of PCR products using primers designed from HTS data (data not shown).The Illumina sequencing data re-analyzed in this work were obtained from the analysis of a selection of individual berries from the Sequence Read Archive (SRA) database of the National Center for Biotechnology Information (NCBI) under the reference PRJNA594635 . The traVirus status of each of the ten individual plants grafted on 5BB was verified by RT-PCR for the eleven viruses detected by HTS. Two plants from the symptomless treatment were missing as they were removed from the collection at the time of sampling (February 2022). Nucleic acids were extracted from dormant wood with a 3% CTAB buffer  following publically available protocol . Primers and PCR conditions are presented in All sequences recovered by mapping with a genome coverage  greater than 94% were deposited on GenBank under accessions ON221453-ON221466 and ON237610. The sequences of GLRaV-1 and GAMaV obtained from both mapping and RT-PCRs were deposited under accessions ON221467, ON221468, and ON583999 .Pseudococcus comstocki was collected from a plum tree in a commercial orchard in Swiss orchards  and a colony was started and maintained on potato tubers (Solanum tuberosum) in jars. Given that mealybug species might be difficult to distinguish based on their morphology, DNA barcoding identification was performed: Genomic DNA was extracted using the DNeasy extraction kit (QIAGEN) and a part of the mitochondrial COI gene was amplified and sequenced according to Correa [In summer 2017, one individual mealybug o Correa . Virus tSymptoms observed in source vines were reproducible in our field experiment . Visual In 2018, sugar content and weight were assessed in single berries at the onset of veraison (when first visible signs of color change occurred). Sugar was significantly lower for the modality GLD_S++ when compared to the symptomless control or to GLD_S+. This sugar drop was not observed between GLD_S+ and the control .The seasonal evolution of leaf chlorophyll content is shown in −2 s−1 over the three seasons, from June to September. All measurements on GLD_S++ leaves showed a significant reduction in AN and transpiration rate (Net photosynthesis (AN) was assessed by measurements of gas exchange, transpiration and stomatal conductance over six days during three seasons . In leavion rate . This reVegetative growth was evaluated by measuring pruning weights and trunk diameter based on rootstock and viral status. Statistical analysis indicated that there was no significant viral treatment x rootstock interaction for vine vigor. However, we observed a tendency of a stronger effect on vine vigor for the treatment GLD_S++ on rootstock 3309C compared to 5BB . PruningFor trunk diameter and mortality evaluated at the end of the study, the data showed a more severe effect for GLD_S++ on rootstock 3309C compared to 5BB . In GLD_Fruit composition and yield at harvest were affected in each of the three seasons by leafroll disease . TreatmeVitis rupestris St-George indicator. This was the only disease detected from the symptomless control. Leafroll symptoms were recorded on samples GLD_S+ and GLD_S++ with early reddening and rolling of the Gamay indicator. Lastly, the fleck symptoms were recorded in the GLD_S+ samples on the indicator Vitis rupestris St-George, with a decreased vigor and vein clearing  analysis. For the bioassay, disease symptoms on indicators were evaluated during three seasons after graft inoculation. The presence of rupestris stem pitting was visible on the three treatments, with pitting and grooving observed under the bark of the clearing . The SRAclearing .The presence of these viral species derived from the SRA was confirmed using specific RT-PCR analysis on each individual vine, and the results are presented in In an attempt to disentangle the different viruses present in GLD_S++, mealybug transmissions were performed. Eight recipient Pinot noir plants were screened for GLRaV-1, GLRaV-3, and GVB infection. Healthy control vines used in transmission assay tested negative for all viruses. Three recipient plants were found to be infected with GLRaV-3 including one mixed-infection with GLRaV-3 and GVB. No virus was transmitted in the remaining five recipient plants. GLRaV-1 was not detected. There was no difference in symptoms expression between the plant mixed-infected with GLRaV-3 and GVB and the two other infected by GLRaV-3. Furthermore, symptoms on these three plants were less severe than the original viral source GLD_S++ .In this study, we examined the impact of leafroll disease on 17-year-old field-grown pinot noir in complement to the previously published transcriptomic analyses. The inoculum GLD_S++ was selected in this experiment because it consistently displayed one of the strongest leafroll symptoms amongst about 900 virus-infected Vitis accessions held in the Agroscope collection, of which at least a third contained one of the five main GLRaVs from different origins present in single or multiple infection . The GLDThese radical differences in symptomology were reflected by three highly distinct viromes, only sharing the two viroids (HSVd and GYSVd) and two viruses (GRSPaV and GRGV). However, with the notable exception of GLRaV-1, strong genetic variability was observed within species sequences obtained from different treatments . It is important to note, however, that these sequences were assembled from a pool of plants and only the dominant sequence or sequences were recovered. Virome homogeneity within a treatment was confirmed by RT-PCR on individual plants with the exception of GPGV and GAMaV only detected on a few plants and, to a lesser extent, GRGV and GSyV-1 detected in most but not all plants . From thP. comstocki. This similarity of symptoms suggests that the severe symptomology of GLD_S++ is caused by a synergy between GLRaV-1 and GLRaV-3 with a possible effect of GVB. However, a specific interaction between the viruses and the rootstock (here 3309C or 5BB) cannot be ruled out, since the recipient Pinot noir plants in mealybug transmission were own-rooted (no rootstock).Early interveinal reddening of the leaves observed on the red cultivars is the first symptom of GLD, followed by downward leaf rolling. It is interesting to note that GLD_S++-infected Pinot noir plants were consistently the first vines of the Agroscope collection to express those leafroll symptoms and more intriguing is that symptoms were observed as early as six weeks prior to veraison. Leafroll-infected plants are known to only express symptoms after veraison and that the timing of the symptom is not only dependent on the inoculation time but also the virus strain in the case of GLRaV-3 infections ,19. PlanSimilarly, we cannot exclude the impact of less studied viruses such as GVT for which its presence is much more likely be unnoticed, in addition to a known pathogen (GLRaV-3). In this respect, it is interesting to note that the increase in the number of virus species detected per treatment  was reflected by an increased percentage of virus reads in total sequencing, estimated around 0.35% for the symptomless treatment, 0.67% for GLD_S+, and 1.84% in GLD_S++ . MoreovePreviously, transcriptome analysis of those plants had shown an upregulation of the plant-innate immunity . The obsOn the other hands, all the assessed parameters from GLD_S++ were severely altered compared to the control . The lower sugar content and berry weight indicate a delay in the onset of veraison for the severe form of GLD; thus, a general delay in berry development is often reported in GLD-infected vines . The negAlthough most of the physiological data were obtained from the rootstock 5BB, vigor and mortality were measured on 3309C and showed an increased disease severity. The rootstock influence on symptoms severity is well documented ,42,43,44Finally, the strength of HTS for plant diagnostic was exposed when we compared the virome identified by sequencing to the results of a three-year long biological indexing assay . The bioP. comstocki. Finally, this study confirms the strength of HTS compared to biological indexing with a faster and more sensitive virus detection. It is important to make good use of this technology to ensure the sanitary status of the planting material as it is, together with the vector control, our only guarantee for a healthy vineyard.To conclude, our study confirmed the serious impact caused by GLD on the plant fitness and wine quality and that severity can be altered by the rootstock selected. In all quantitative and qualitative measurements, GLD_S+ vines showed an intermediate response to the disease between symptomless and GLD_S++, making it a good gradient model for the virus response, but the complexity of the viromes identified did not allow the severity of symptoms to be attributed to the addition of a single virus but rather to a synergy between the viruses present. This research reports the presence of GAMaV for the first time in Switzerland and the first transmission of GVB by


The first herpes virus to be described was types 1 and 2, whose denomination is herpes simplex 1 and 2 or HSV -1 and HSV -2. These viruses have specific biological characteristics, such as the ability to cause different kinds of diseases, as well as to establish host's latent or persistent lifetime infections and also of being reactivated, causing lesions that can be located at the same site of the initial primary infection or close to it. It is suggested that this virus reactivation in the geniculate ganglion may be related to Bell's palsy. In this situation, the viruses that would be latent in this ganglion, would suffer reactivation and replication, then be diffused through the facial nerve and its branches, among them the chorda tympani nerve, which by stimulating salivary secretion would enable the identification of the viral DNA in the patients’ saliva. Until recently, a great number of patients was diagnosed as holders of this kind of paralysis, named idiopathic or Bell's palsy. With the introduction of the technique studying the viral DNA by Polymerase Chain Reaction (PCR), several authors have found herpes simplex virus type I DNA in the cerebrospinal fluid, in the lachrymal secretion, in the saliva and in the geniculate ganglia of patients with Bell's palsy.observe the occurrence of herpes simplex type I virus using PCR technique in the saliva of patients with Bell's palsy and relating it to the clinical evolution of these cases.We evaluated 38 patients with Bell's palsy submitted to anamnesis, clinical and ENT examination and saliva sampling for viral DNA detection by PCR technique. The control group was ten normal adults.We found positive viral DNA in 11 cases out of the 38, which corresponded to 29% of the sample. This result was statistically significant if compared to the control group, in which we did not find any positive case.The end result was that the presence of HSV -1 in the saliva of patients with Bell's palsy indicating that the viral reactivation can be the etiology of this disease. The detection of the virus in these patients’ saliva does not influence the disease prognosis. Types 1 and 2 were the first herpes virus described (HHV-1 and HHV-2), called herpes simplex 1 and 2 or HSV-1 and HSV-2. These viruses bear particular biologic characteristics, such as the potential to cause different types of diseases, to establish latent or persistent infections during the host's life time and that of being reactivated causing lesions that may be located on the site of the primary initial infection or near it.The lesions caused by these viruses were first documented by Hypocrates (460/377 b.C.), who called them herpes, from reptile, because of the skin vesicles. But it was only in 1968, that two types of herpes virus - Simplex virus genus were classified on the basis of their biological and antigenic differences. These observations were fundamental for clinical, serological, immunological and epidemiological studies that peaked at the establishment of the anti-viral therapy, and it also demonstrated the genotypic and phenotypic differences among variants of HHV-1 and 2.The International Committee on Taxonomy of Viruses (ICTV) classified this virus in the Herpes viridae family, Alphaherpesvirinae subfamily, Simplex virus genera, types 1 and 2. Both HSV-1 and HSV-2 are made up of an electrodense core of 150-153 Kpb double linear DNA strains. HSV-1 and 2 infect different cell types, such as fibroblasts, squamous and mucous cell epithelia, polarized cells of the cylindrical epithelium, glial cells and nerve endings.These virus show the capacity to establish latent or persistent infection throughout its host's life, changing the latency site according to its subfamily. The major characteristics of the alfa-herpesviridae family members is to establish latency in the sensory nerves. During this latency period, the viral DNA is not completely silent. None of the viral genes expressed during the lytic phase is detected. However, some RNA called latency associated transcripts may be found in high levels. In the same way, these viruses may show the reactivation mechanism, when the virus may be reactivated in peripheral tissues, and this could, at least in part, explain the virus transmission in an asymptomatic period.Clinically, most HSV-1 is isolated in herpetic lesions of the oropharynx. After the acute or symptomatic phase of virus manifestation, it may remain latent in the sensitive ganglia neurons, as in the facial nerve case, the geniculate ganglion. The viral reactivation in this site may cause an inflammatory process able to trigger a peripheral facial paralysis. This is the main disease that affects the facial nerve and has broad etiology, where systemic, inflammatory and tumoral diseases, among other, may determine itClinically the peripheral Facial Paralysis is a disease characterized by the loss of muscle movement in half the face, generating a clear deformity during facial expressions.Until recently, a large number of patients was diagnosed as carriers of one type of this paralysis, the one called idiopathic or Bell's paralysis. These terms are still used in the literature to characterize those patients with acute signs of peripheral facial paralysis and without an etiological diagnosis defined even after conventional clinical, laboratorial and image investigationsStarting on the 70's, many studies tried to establish this disease etiologyIdentifying HSV-1 DNA virus in all these circumstances indicate that the type 1 herpes simplex virus may very well be the major agent responsible for this peripheral facial paralysis (PFP)The goal is to detect type 1 herpes simplex virus by PCR in the saliva of Bell's Peripheral Facial Paralysis patients, comparing them to a control group and correlating it to electroneurography and degree of paralysis results.The study was carried out in a tertiary University Hospital and was referred and approved by that institution Medical Ethics Committee.a)Acute peripheral facial paralysis with symptoms starting up to seven days after they were first seen at the hospital;b)Physical and otolaryngological exam without evidences of a determined cause for the PFP.38 consecutive patients were assessed and diagnosed as carriers of Bell's Peripheral Facial Paralysis according to the following inclusion criteria:The control group encompassed 10 normal adults.a)Medical follow upb)Material harvestThe patients were seen as they arrived at the medical facility, were then interviewed and went through general and otolaryngological physical exams.c)Lab examsIn this visit, saliva was harvested from the patients’ mouth using a disposable sterile syringe without the needle. The saliva was stored in a proper container for the PCR (EPENDORFF).d)Statistical analysisTests were carried out in order to detect the HSV-1 viral DNA through PCR using specific primers aforementioned for this virus.The results obtained both from the clinical and laboratorial views were statistically analyzed.38 patients with acute Bell's PFP were assessed, 16 males and 22 females. Of those, 11 (29%) had positive PCR for the HSV-1 virus in the first week of disease development. The positive PCR cases were divided in 5 females (23%) and 6 males (37%).The age ranged from 6 to 71 years, and 12 of these patients were between 21 and 30 years. We could see that in the assessed clinical cases, the stapedial reflex was not present in 21 cases and present in the other 13 cases. Among PCR positive cases, 9 patients (80%) did not have this reflex while only 2 patients (20%) had the reflex present.As far as development is concerned, of the 38 patients assessed, 27 were discharged without sequelae and three remained in grade II of the House-Brackmann classification. Two remained in grade V, three patients were decompressed and three did not have clinical follow up.Of the positive patients, three did not improve and one was referred to surgery, another remained in grade II and another, still, in grade V.In the acute PFP the medical team has to act quickly in order to establish the most adequate diagnosis and treatment for the patient and thus, avoid future facial motor sequelae. It is of fundamental importance to define the etiology both for treatment purposes and to provide a more accurate prognosis. It is not always easy though, therefore it is important to carry out a careful clinical evaluation and, in many cases, a number of complementary tests.Bell's palsy in our settings is characterized for being a diagnosis of exclusion. A new situation is arising with modern lab techniques for viral investigation that is making this current concept, which is around since the 90's, broadly modified and the facial nerve involvement in the PFP is better assessed.In this study, 29% of the Bell's PFP cases had HSV-1 positive test in their saliva. This figure is greater than the one found by Furuta et al., who evaluated 176 patients. Compared to the results from Abiko et al., our data coincide, although his sample was smaller (16 patients). We may believe that these positive cases may have had a virus reactivation process as etiology.The establishment of this relationship may decisively influence the introduction of antiviral drugs in the treatment of these patients. HSV-1 was not influenced by the age factor in our investigation.This finding is not broadly discussed in the literature and does not uphold the idea that viral reactivation occurs more frequently in elderly patients. One relevant fact was that we did not find the stapedial reflex in 80% of HSV PCR positive cases. PCR negative cases had equal distribution as far as the presence or absence of this reflex is concerned.Analyzing this fact, we can postulate that if the virus is reactivated in the geniculate ganglion, migrated through the facial nerve and reached the salivary gland (via chorda tympanis nerve), it is likely that there may have occurred an inflammatory process in the facial nerve tympanic-mastoid segment and, with that, involvement of its stapedial branch (reflex absence) happening more frequently.As to facial nerve electroneurography, we did not observe significant difference of results in comparison with PCR positiveness for HSV-1 . TherefoAs to PFP degree, we observed that negative PCR for HSV-1 had a homogenous distribution among grades II to IV. In positive PCR cases, 63% presented grades V or VI. This would indicated that the virus causes greater neural involvement. However, such data is not corroborated by facial nerve electroneurography findings.Another important fact is patient clinical evolution, assessed according to HSV-1 positiveness. As we observed, positive or negative PCR HSV-1 did not influence patient recovery. This fact was also observed by Furuta et al.HSV-1 present in the saliva of Bell's PFP patients tells us that viral reactivation may be the very etiology of this disease. Detecting the virus in the saliva does not alter the disease prognosis.


The Internal–External Locus of Control Short Scale–4 (IE-4) measures two dimensions of the personality trait locus of control with two items each. IE-4 was originally developed and validated in German and later translated into English. In the present study, we assessed the psychometric properties  of the English-language IE-4, compared these psychometric properties with those of the German-language source version, and tested measurement invariance across both language versions. Using heterogeneous quota samples from the UK and Germany, we find that the English-language adaptation has satisfactory reliability and plausible correlations with 11 external variables , which are comparable with those of the German-language source version. Moreover, metric measurement invariance of the scale holds when comparing the UK and Germany, implying the comparability of correlations based on the latent factors across the two nations. As an ultra-short scale (completion time < 30 s), IE-4 lends itself particularly to the assessment of locus of control in survey contexts in which assessment time or questionnaire space are limited. It can be applied in a variety of research disciplines, such as psychology, sociology, or economics. Locus of control is defined as a generalized expectation of internal or external control of reinforcement . IndividInternal and external locus of control are predictive of a variety of behavioral, cognitive, and affective outcomes in different areas of life, including well-being, satisfaction, and performance-related behavior and outcomes . StudiesAn empirical validation of the English-language IE-4 was hitherto lacking. The aim of the present study is to fill this gap. Specifically, first, we analyzed the reliability and factorial structure of the scale, and its correlations with a broad range of 11 external correlates in order to validate the scale and to embed it in a nomological network. Second, we compared these psychometric properties with those of the German-language source version. And finally, we investigated measurement invariance across the two language versions. For this purpose, we used heterogeneous quota samples of adults from the United Kingdom (UK) and Germany (DE).The theoretical background section of this paper is based on, and parts of it are taken from, Kovaleva et al. . The conIn his social learning theory, Rotter , 5 assumLocus of control also explains several other phenomena within and across individuals, thereby rendering it an important construct for a variety of research questions. It has been related to a variety of behavioral , affectiFor instance, a plethora of research indicates that individuals who have a more internal locus of control spend more time on intellectual and academic activities compared with externally controlled individuals . ConsequRegarding justice sensitivity, previous research has been reported the opposite direction compared to most other effects with locus of control, namely a negative correlation between internal locus of control and justice sensitivity as well as positive association between external locus of control and justice sensitivity , 27.People with a more internal locus of control have been found to be more willing to take risks , 29, to Locus of control has stimulated a lot of research since its initial introduction by Rotter , 5, and Based on these findings, and aware of the general time constraints in surveys, Jakoby and Jacob  developeGiven the lack of comprehensive validations of German-language locus of control scales such as the KMKB that were suitable for use in surveys, Kovaleva  and KovaFor the original German-language IE-4 scale, Kovaleva et al. . In a seTo enable social scientists to use IE-4 in an English-language context, the scale was adapted to the English language by Kovaleva et al. . Two prodoes not apply at all (1) to applies completely (5). For each subscale, the unweighted mean score of the respective two items is computed to obtain subscale scores for internal and external locus of control. Computing a total mean score across both subscales is not recommended. We suggest that individual answers should be aggregated to the scale level only if there are no missing values on any of the two items. In our two samples (UK and Germany) there were no missing values. If there are missing values, we recommend using appropriate methods for handling missing data, such as multiple imputation ) to 17 . An 18th category  was provided for those who had no income. None of the participants chose that category. Before computing the correlations, we recoded the negatively worded items of all short scales (for both language versions), the subdimension “minimizing negative qualities” of socially desirable responding (for both language versions), and the self-esteem scale (UK only), so that high values always represented high levels of the respective traits. Because the Big Five dimension Emotional Stability is negatively worded in relation to the construct Negative Emotionality in the BFI-2-XS, we recoded the respective items so that high values represented the positive pole of this dimension—that is, Emotional Stability. In addition, we recoded the employment status variable and tested two contrasts: (a) unemployed (out of work and looking for work/out of work but not currently looking for work) versus (self-)employed, and (b) retired/doing housework versus (self-)employed. We did not make further contrasts, and regarded all other employment status categories  as missing values because the sample sizes of these categories were too small.To assess income, respondents were asked to allocate their net income to one of 17 categories ranging from 1 . The code can be found in the r = .12 in the UK and r = −.29 in the German sample) and the inter-item correlations  the administrator (objectivity of application) and (b) the evaluator of the instrument , and when (c) unambiguous and user-independent rules are provided (objectivity of interpretation). The standardized questionnaire format and written instructions, the fixed scoring rules and labeled response categories, and the reference ranges ensured the objectivity of the application, evaluation, and interpretation of IE-4.rtt, over a period of about 28 days (Mdn) in the UK (NUK = 111) and 20 days (Mdn) in Germany (NDE = 117), respectively. Our reasoning was that this time span of 3 to 4 weeks was long enough to allow for meaningful test–retest stability estimates and short enough to preclude the occurrence of pronounced and systematic change in the true scores of internal and external locus of control. Because the test–retest stability is sensitive not only to measurement error but also to state fluctuations in dispositional internal and external locus of control  for internal locus of control and r = .56, 95% CI  for external locus of control.Despite some differences in the patterns of correlations between the UK and Germany, the overlapping confidence intervals suggest that many of these differences were statistically non-significant. Overall, the nomological networks were fairly similar, albeit not identical, across nations. The profile similarities  were 2 difference test is significant and/or ΔCFI ≤ −.010 either in combination with ΔRMSEA ≥ .015 or ΔSRMR ≥ .030; scalar and full uniqueness invariance must be rejected when the χ2 difference test is significant and/or ΔCFI ≤ −.010 either in combination with ΔRMSEA ≥ .015 or ΔSRMR ≥ .010.We assessed the comparability of IE-4 across the UK and Germany via measurement invariance tests with multiple-group confirmatory factor analyses (MG-CFA) , 78. The2(6) = 11.382, p = .077, CFI = .986 (robust CFI = .987), RMSEA = .044 (robust RMSEA = .047), SRMR = .025, BIC = 10,749.117—metric invariance can be accepted, implying the comparability of correlations based on the latent factors between both nations. When comparing the scalar model—χ2(6) = 86.554, p < .000, CFI = .793 (robust CFI = .820), RMSEA = .144 (robust RMSEA = .153), SRMR = .065, BIC = 10,819.123—with the metric model, the significant χ2 difference and the MLR-scaled CFI indicated that scalar invariance of IE-4 did not hold across the UK and Germany: Δχ2(2) = 84.548, p < .001, ΔCFI = –.193 (Δ robust CFI = –.167), ΔRMSEA = .010 (Δ robust RMSEA = .106), ΔSRMR = .004, ΔBIC = –70.006.Because the metric model showed a good fit—χThe aim of the present study was, first, to empirically assess the psychometric properties of the Internal–External Locus of Control Short Scale–4 (IE-4), the English-language adaptation of the German-language source version Internale-Externale-Kontrollüberzeugung-4, developed by Kovaleva et al. [The non-achievement of scalar invariance indicates that the latent means between the UK and Germany are not comparable without systematic bias, implying that the two countries do not have the same point of origin. A possible reasons for this could be, for example, that the two nations used different frames of reference when assessing their locus of control beliefs . To accoWe could replicate the two-dimensional structure of internal and external locus of control in both language versions. As expected, in most cases, these two dimensions were differentially associated with a total of 11 other psychological constructs, which we used to embed the scale in a nomological network. In other words, either the direction of the effects  or the effect sizes differed, or both. Taken together, we could, for the most part, support and expand the findings of the original validation study of the German-language source version  with resIn addition, there were also some discernible differences between the two nations, both in terms of the direction of the effects and of their strength. In the UK, we found, for example, a small positive effect between external locus of control and scores on the impulsive behavior subscale perseverance, whereas in Germany we found a small negative effect between the two variables. Moreover, there were medium-to-large-sized positive associations between both locus of control dimensions and external political efficacy in the UK, whereas there were zero correlations in Germany. By contrast, we found a similar pattern for the relation between internal locus of control and Agreeableness, but in this case with a medium-sized positive effect in Germany and a zero correlation in the UK.The size of the factor loading of Item 4 was also different between samples, and was twice as large in the UK as in the German sample. Furthermore, the correlation between internal and external locus of control also differed across nations: whereas in the UK, there was just a medium-sized positive association, the association in Germany was negative and twice as large as that in the UK. All these differences may be due to national specificities, such as differing socialization. However, this is purely speculative; further research is needed to explore these cross-national differences in depth. Moreover, future studies could also examine correlations between IE-4 and more fine-grained subdimensions/facets of each construct in order to further elaborate the nomological network of IE-4. In doing so, a network analysis could be performed to show the graphical representation of the correlation matrix.r = .92–.99) [Despite the benefits of our study, its scope was limited in three ways. First, both samples were restricted to participants in a Web-based survey (CASI); second, the English-language sample was restricted to the population of the UK. Consequently, we cannot generalize our findings to the whole population—including, for example, persons who are not computer literate—or to other English-speaking populations, for example, in the United States. Although there is no reason to expect major differences between survey modes or English-speaking nations, future research might address these issues. Third, due to survey time constraints , we could not include alternative measures of internal and external locus of control in our study. However, previous research has reported that the two dimensions of the German-language source scale of IE-4  correlated highly with the corresponding dimensions of another locus of control scale, KMKB , which h.92–.99) , indicatTo conclude, the results of the present validation study show for the first time the utility and psychometric properties of the English-language adaptation of the ultra-short IE-4 scale and the comparability of its psychometric properties with those of the German-language source version. Researchers in English-speaking nations now have the possibility of measuring internal and external locus of control in an economical and time-efficient way in assessment settings with limited resources, such as large-scale surveys in the social sciences. We recommend applying IE-4 only in self-report surveys for research purposes in measurement settings with severe time limitations, and not for individual diagnostics.S1 AppendixIE-4.(PDF)Click here for additional data file.S2 AppendixIE-4.(PDF)Click here for additional data file.S3 Appendix(PDF)Click here for additional data file.


In Germany, recommendations on infection prevention and control of current virus outbreaks are given as communications by the Association for Applied Hygiene e.V. (VAH) together with the joint Disinfectant Commission of the German Association for the Control of Virus Diseases e.V. (DVV) and the Society of Virology* (GfV). The DVV was founded in 1954 in response to the ongoing threat to the population from polio and was given its current name in 1977. The DVV is supported by the Federal Ministry of Health, the Ministries of Health of the Federal States, scientific societies, as well as social foundations and organisations. Private individuals cannot be members of the DVV. The Society of Virology e.V. (GfV) is a scientific society for all virological fields in Germany, Austria and Switzerland, and is thus the largest virological society in Europe. With numerous commissions, guidelines and statements, it is the authoritative contact for research, healthcare and politics. The joint commission “Virus Disinfection” of these scientific societies focuses on the efficacy of chemical disinfection procedures against viruses. The VAH bundles the expertise of scientific societies and experts on infection prevention and is particularly committed to the quality assurance of hygiene measures. With the VAH disinfectant list, the association provides the standard reference for the selection of high-quality disinfection procedures. This disinfectant list has a tradition of more than 60 years in Germany.The original German version of this document was published in August 2022 and has now been made available to the international professional public in English. The document contains recommendations on hygiene and disinfection measures for monkeypox virus infections. Disinfectants against monkeypox must have at least proven efficacy against enveloped viruses (active against enveloped viruses); products with the efficacy ranges “limited virucidal activity” and “virucidal” can also be used. The disinfectant list of the VAH or the disinfectant list of the Robert Koch Institute are available for the selection of products. Especially in the case of contamination with crust or scab material, it should be noted that protein contamination can have a protective or stabilising effect on monkeypox. Therefore, cleaning – before disinfection – should always be carried out in this situation. Preventive measures such as vaccination and hygiene in the vicinity of people with monkeypox must be taken to prevent transmission to small children, pregnant women or people with a pronounced immune deficiency. Poxviridae family. The genus of orthopoxviruses also includes, among others, the variola virus , the vaccinia virus, the horsepox virus and the cowpox virus  as a vaccine for the first time. The procedure called "vaccination" quickly became established. In addition to the variolation of cows or human-to-human vaccination, other animals such as rabbits, pigs, sheep, donkeys, horses and goats were also used as intermediate hosts to improve the effectiveness of the lymph. In fact, vaccines derived from cowpox or horsepox were used interchangeably for smallpox vaccination in the 19th century ; t; t13]; tThe WHO has established an Emergency Committee and published recommendations for action to protect public health. It also reported cases of infected children in the United Kingdom, in Spain and France with mild courses . Monkeypox is a rare but potentially serious viral disease that typically begins after an incubation period of 5–21 days with a flu-like illness , headache, muscle pain and fatigue) and swelling of the lymph nodes  and develops into a rash on the face and body.The eruptive stage begins with typical enanthema (oropharynx) and exanthema on the face, hands, forearms with centripetal spread over the body and subsequent development of redness and pox-typical uniform efflorescence stages . This occurs in about 80% of patients within a few days – 20% of sufferers develop polymorphous exanthema– similar to varicella. The lesions heal after drying and desquamation (sometimes with scarring). It was previously assumed that the infectivity lasts from the beginning of the prodromal stage at least until the crust of the skin lesions falls off . People vaccinated against smallpox generally develop fewer efflorescences than non-vaccinated persons. In addition, ulcerations on the mucous membranes of the oral cavity with pharyngitis and tonsillitis, conjunctivitis with eyelid oedema and very painful lesions in the genital area frequently appear in non-vaccinated persons. Rarely, blindness and disfiguring scars occur as permanent damage. Severe, fatal haemorrhagic forms are rare; mild forms with less than 10 pockmarks and subclinical infections are sometimes observed.Overall, the prognosis can be considered favourable. A higher probability of severe disease and mortality has only been observed in children under 8 years of age in the past , 17]. . 17]. Recent reports show that the symptomatology of the monkeypox clade 3 diseases now prevalent in Europe differs from earlier descriptions. Symptoms of the prodromal stage are often absent. The skin lesions first appear in the urogenital and anal regions and not on the hands and soles of the feet, as is more common. The lesions were also often at different stages and may have occurred before systemic symptoms. The team of authors of a study published in Lancet Infectious Diseases in July 2022 therefore suggest an adjustment of the case definition .Asymptomatic courses are also reported in a preprint study from Belgium. In anorectal swabs, a positive result could be detected in three (1.3%) of 224 MSM retrospectively tested for monkeypox. All those affected stated that they had not had any symptoms .The possibility of generalised and severe courses in the case of impaired immunity  should be taken into account. Therefore, it is also important to provide special protection for these population groups.When reporting vaccination status, it is advisable to record the vaccination status from the vaccination record. A study by Eurosurveillance found that positive vaccination status against smallpox virus was also reported by patients under 40 .Monkeypox virus can be transmitted via different routes, namely animal-to-animal (predominantly different rodent species), animal-to-human, and human-to-human. In addition, the possibility of human-to-animal transmission cannot be ruled out in the case of contact with high virus loads. Monkeys as well as humans are false hosts for the monkeypox virus . In human-to-human transmission in the current outbreak, the focus is on direct transmission via close contact with infectious skin lesions (e.g. via ruptured blisters). The blisters and pustules contain high viral loads. Ports of entry are small skin lesions as well as the mucous membranes and the respiratory tract. In addition, indirect transmission via infectious material  is possible. Vertical transmission from mother to child has also been described in rare cases , 23]. T. T23]. Tth day of accommodation of monkeypox patients. By means of PCR analysis, it was found that the surfaces of the wet cells close to the patients  had a particularly high viral load, as did seating surfaces of chairs and the display of the patients’ mobile phones, as well as textiles  used by the patients. Furthermore, surfaces that were presumably touched by medical staff and thus contaminated, such as cupboard handles, door handles of the anteroom, showed high viral loads. The authors restrictively point out that these are mainly results of a PCR analysis, i.e. viral DNA, and not the cultivation of infectious monkeypox viruses. Interestingly, however, they were able to culture monkeypox virus in three of the samples collected from one patient, namely from the investigator's glove, the operating lever of the soap dispenser and a towel on the patient's bed. All three samples had more than 106 copies per sample (>103 cp/cm2) , , . In the EU, the smallpox vaccine Imvanex is licensed against smallpox, which contains a modified form of the vaccinia virus Ankara (MVA) that is no longer able to replicate. In the U.S. and Canada, the approval of this vaccine also extends to vaccination against monkeypox. In the European Medicines Agency (EMA), the review of data on the indication extension of Imvanex has started . AccordiIn the meantime, the vaccine is available in the practices in Germany, and vaccinations according to the STIKO recommendations have started. The organisation and vaccination is regulated by the federal states.The most important non-pharmaceutical preventive measure for the further spread and disease of monkeypox is the avoidance of close contact with an infected person. Patients and also the persons living in the same household with a monkeypox patient should be advised by a doctor and, if possible, trained on which hygiene measures to take and how to carry them out properly Table 1 .“virucidal activity against enveloped viruses” , , . T. T“virucnt virus , 45]. . “viruci“limited spectrum of virucidal activity” and “virucidal activity” can also be used . V. V47]. V“limited spectrum of virucidal activity” or “virucidal activity” would only be discussed  if, due to their mechanism of action, they could penetrate crusts/scabs and also inactivate the virus inside, if prior efficient cleaning of the surfaces is not possible. The use of disinfectants with the effective range Laundry disinfectants are an exception here, as the European standardisation and also the VAH and RKI lists only specify the virucidal range of action. If no VAH certificate is available, it is recommended that an evaluation of the test reports and expert opinions submitted by the manufacturer be carried out by independent experts , 49]. . 49]. The criterion for the selection of a disinfectant should not be a specific active ingredient or group of active ingredients, but the manufacturer-independent proof of efficacy for the required spectrum of activity for a specific product. The importance of laundry preparation, including the preparation of mopping utensils, for instance, for floor cleaning (mops), should be emphasised. Care must be taken that bed linen and body laundry is collected in such a way that, as far as possible, there is no environmental contamination with skin crusts, as the viruses embedded in these are much more difficult for disinfectants to reach. Chemo-thermal reprocessing with a virus-effective, VAH- or RKI-listed procedure is required for laundry reprocessing in the clinical and nursing environment . Disinfectants against monkeypox must have at least proven efficacy against enveloped viruses (“active against enveloped viruses”); products with the efficacy ranges “limited virucidal activity” and “virucidal” can also be used. The disinfectant list of the VAH or also the disinfectant list of the Robert Koch Institute are available for the selection of products . EspeciaThe authors declare that they have no competing interests.This recommendation is also available in German:https://vah-online.de/de/ and https://g-f-v.org/komissionen/Eggers M, Exner M, Gebel J, Ilschner C, Rabenau HF, Schwebke I. Joint communication from VAH and the Virus Disinfection Commission of the DVV and GfV: Wirksamkeit von Desinfektionsmitteln gegen Affenpockenviren. Status 2022 Jul 18. HygMed. 2022;47(7-8):158-64. Available from:


There is empirical evidence of an association between periodontitis and coronary heart disease (CHD). However, it is uncertain whether periodontal treatment in CHD patients might lead to reduced healthcare costs. This study aims to assess the association between periodontal treatment and healthcare costs in newly diagnosed CHD patients. Data from 21,263 adults who were continuously insured between 2011 and 2016 and who were newly diagnosed with CHD in 2013 were selected from a German claims database. The study population was differentiated by the utilization of periodontal treatment. The average treatment effect (ATE) of periodontal treatment on healthcare costs  was investigated using weighted Poisson regression models conditional on covariates and is shown as a ratio (of geometric means). Periodontal treatment was documented for 4.7% of the persons in the study population. Newly diagnosed CHD patients showed an ATE of 0.98 for total healthcare cost (95% CI 0.90–1.06), 0.79 for inpatient costs (95% CI 0.61–1.04), and 0.95 for drug costs (95% CI 0.87–1.04). A statistically significant 7% increase in outpatient costs was shown (95% CI 1.01–1.13). Despite a lack of statistical significance in most cases, the study provides evidence of a meaningful decrease in inpatient costs after periodontal treatment. Further studies are needed. Periodontitis and coronary heart disease (CHD) are highly prevalent non-communicable diseases ,2. In 20CHD and periodontitis are two multifactorial diseases with common risk factors, such as diabetes or smoking ,8. FurthAgainst the background of high prevalence and an aging population, both periodontitis and CHD cause a high financial burden for the healthcare system ,21. WhetTo this end, the present study investigated the association between periodontal treatment and different kinds of healthcare costs in patients newly diagnosed with CHD. We hypothesized that periodontal treatment will significantly reduce healthcare costs in patients with CHD.We conducted a retrospective cohort study. The full dataset of the InGef  research database was used. The InGef research database includes longitudinal health insurance data on at least six million Germans insured with about 70 statutory health insurers, mainly company health insurance providers . Data foThe plausibility check showed negative or missing costs in the inpatient sector. Costs were imputed for the corresponding inpatient cases according to the following procedure: Based on inpatient cases from the entire InGef research database for which cost data were available, mean costs were calculated per year and per Diagnosis Related Group or, if no Diagnosis Related Group code was available, per treatment type for one hospital day. For inpatient cases with missing/negative costs the calculated mean costs were extrapolated to the respective length of stay.The study population comprises individuals who were continuously insured between 2011 and 2016, were at least 18 years old in 2013, and were newly diagnosed with CHD in 2013 . CHD wasThe study population was subdivided into an exposed and unexposed group. The exposed group included all individuals from the study population who had received at least one periodontal treatment in the index quarter or the eight subsequent quarters. The unexposed group was defined as having had no periodontal treatment in the index quarter or the following two years. The treatment fee codes P200-P203, 108, and 111 according to the German uniform assessment standard for dental services (BEMA) were used to determine the periodontal treatments  26]..26].In this study, total healthcare costs from the viewpoint of statutory health insurance are used as the outcome and defined as the sum of inpatient, outpatient, and drug costs. Costs are expressed in Euro. The costs of each sector individually were also considered. In the outpatient sector, only costs from the area of human medicine, i.e., excluding dental costs, were taken into account.Potential confounders of the association between the utilization of periodontal treatment and healthcare costs were identified as health status, motivation to seek healthcare, and area of residence, in addition to age (in the index quarter) and sex. Based on the available data, the variables were operationalized as follows: Health status via Charlson comorbidity index, and motivation to seek healthcare via (i) total healthcare costs one year before the index quarter , (ii) number of physician groups visited one year before the index quarter (excluding dentists), and (iii) dental visits (yes/no) one year before the index quarter .The information on area of residence was available at the federal state level. Due to low case numbers in some federal states, a regional allocation to Nielsen regions was made . The ChaTo examine the association between periodontal treatment and healthcare costs, the average treatment effect (ATE) was computed. A counterfactual framework was applied. Two potential outcomes are assumed for each individual; one with and one without periodontal treatment. However, in this cohort study, an individual can only be assigned to the exposed or unexposed group. That is why the counterfactual outcome of individual A was defined by searching the dataset for an individual B with comparable characteristics and different treatment to individual A. The exchangeability of the individuals is assumed . The difDescriptive and multivariate analyses were performed. As the main multivariate model, a doubly robust (DR) method was chosen. The DR method builds on the combination of outcome regression and inverse probability weighting methods. Based on a weighted Poisson regression model conditional on the covariates for both the exposed and unexposed group, the predicted values for the whole sample were obtained . The meaFor multivariate analysis, all costs were log transformed as a result of a right-skewed distribution (log (cost + 1)). For better understanding, the ATE results were back-transformed by an exponential function and displayed as a ratio (of geometric means). For comparison, a simple linear regression, an inverse probability weighted Poisson regression, as well as a Poisson regression conditional on the covariates were also conducted. The significance level alpha was set to 0.05 for all analyses. All analyses were performed using SQL and SAS 9.4 .n = 1003) of study participants could be assigned to the exposed group. Differences in exposed and unexposed group are most apparent with respect to dental care utilization before the index quarter and total healthcare costs before the index quarter. The exposed group visited dentists more often and had lower healthcare costs before the index quarter  is presented in This study analyzed the association between periodontal treatment and different kinds of healthcare costs in newly diagnosed CHD patients. Statistically significant results were only seen in outpatient costs. A slight increase in outpatient costs after periodontal treatment was evident. In contrast, a tendency towards more decreasing costs in the inpatient sector was observed, but without statistical significance. Our study is the first to investigate this topic based on German data in a population of CHD patients. However, the findings of our study cannot confirm the increase or reduction in total healthcare costs after periodontal treatment observed in two previous claims data studies. The US study by Jeffcoat et al. demonstrated a 10.7% annual reduction in medical costs (inpatient and outpatient) . In contFor the study by Albert et al. , it is lFurthermore, Jeffcoat et al. included only individuals with at least one periodontal treatment as diagnosed by a clinician. That means that patients with completed periodontal treatment  were compared with patients with three or fewer periodontal treatment visits (control group) . In contBesides reducing medical costs, Jeffcoat et al. demonstrated a 28.6% decrease in hospital admission rates in patients with CHD and periodontal treatment . A cohorTo our knowledge, the impact of periodontal treatment on outpatient or drug costs explicitly in CHD patients has not been investigated by other studies. While our data show a slight tendency towards decreasing drug costs in the absence of statistical significance, a reverse and statistically significant effect was seen for outpatient costs. Lower drug costs after periodontal treatment could be explained by the less frequent occurrence of adverse cardiac events and the positive effect of periodontal treatment on the whole complex of metabolic disorders , which cOur study confirms that compared to the high prevalence of periodontitis in Germany , especiaIf periodontitis is detected at an early stage, the burden of disease could already be reduced by regular professional tooth cleanings and adequate individual oral hygiene . The denOur study has strengths and limitations. Claims data have the advantage of providing a large and longitudinal dataset that allows for the cohort study design, and rare events can also be detected . FurtherIn contrast, the following limitations must be considered. First, our study is based on data mainly from company health insurance policies and the study population was restricted to patients continually insured between 2011 and 2016. That means that individuals who changed health insurance policies during this period  or died  could not be considered. Second, claims data are collected for billing purposes. In Germany, a considerable proportion of professional tooth cleaning is being provided outside of statutory health insurance coverage and such utilization is not included in our claims data. Professional tooth cleaning may already be sufficient for the treatment of mild periodontitis . EvidencThird, the oral health status of individuals is not available in German claims data and so neither the diagnosis of periodontitis nor information regarding the severity of the disease was available to us. However, the lack of diagnosis is not a major problem in this study, as the costs of periodontal treatment are only covered by the statutory health insurance if there is evidence of a periodontal disease. Given the high prevalence of periodontitis in Germany and the known association between periodontitis and CHD ,10,11,12Fourth, our study assessed outcome costs in the third year after the initial diagnosis of CHD. In the two previous years, the utilization of periodontal treatment was classified. We hypothesize that the elimination of periodontitis has a protective cardiovascular impact in our observation period, via the mechanisms described above. Nevertheless, it is possible that a longer period between exposure and outcome would reveal relevant cost effects due to fewer adverse cardiac events. This should be considered in further studies. Finally, because of the “small” exposed group and the imbalance between the exposed and unexposed groups, we may have failed to demonstrate statistically significant treatment effects in most cases.Due to the limitations, our findings can only be generalized with caution.In conclusion, due to a low utilization of periodontal treatments by insured persons, the results of our study are not sufficiently robust. In most cases, no statistically significant effects could be demonstrated. Prospective studies with higher numbers of exposed cases are needed. However, our study provides evidence of meaningful decreasing inpatient costs after periodontal treatment.


Rhipicephalus ticks in Guangdong, southern China to identify RNA viruses. Meta-transcriptome analysis revealed the virome in Rhipicephalus ticks, resulting in the discovery of 10 viruses, including Lihan tick virus, Brown dog tick phlebovirus 1 and 2 in the family Phenuiviridae, Mivirus and Wuhan tick virus 2 in the family Chuviridae, Wuhan tick virus 1 in the family Rhabdoviridae, bovine hepacivirus in the family Flaviviridae, Guangdong tick quaranjavirus (GTQV) in the family Orthomyxoviridae, Guangdong tick orbivirus (GTOV) in the family Reoviridae, and Guangdong tick Manly virus (GTMV) of an unclassified family. Phylogenetic analysis showed that most of these TBVs were genetically related to the strains in countries outside China, and GTQV, GTOV, and GTMV may represent novel viral species. These findings provided evidence of the long-distance spread of these TBVs in Guangdong, southern China, suggesting the necessity and importance of TBV surveillance.Tick-borne viruses (TBVs) have increasingly caused a global public health concern. This study collected  Flaviviridae and Crimean-Congo hemorrhagic fever virus (CCHFV) in the family Nairoviridae are responsible for encephalitis and hemorrhagic fever in northeastern and northwestern China, respectively. Emerging TBVs, such as severe fever with thrombocytopenia virus  are considered to be next to mosquitoes as vectors for pathogenic agents. Tick-borne viruses (TBVs) include a large group of viruses of at least 12 genera, nine families, and two orders . In Chinia virus , Jingmenia virus , Alongshia virus , Songlinia virus , Beiji nia virus , and Tacia virus , have bein China .Rhipicephalus sanguineus and Rhipicephalus microplus ticks in Guangdong, southern China using metagenomics and detected 10 viruses, including 3 novel viruses, suggesting a high viral diversity in Rhipicephalus ticks.High-throughput sequencing technology has been widely used to investigate viromes in ticks and diagnose unknown infectious diseases. To date, tick viromes have been reported in Heilongjiang, Liaoning, Hebei, Henan, Hubei, and Yunnan provinces of China , revealig for 5 min at 4°C. Supernatants were collected for RNA extraction with TRIzol LS reagent .From May–June 2020, ticks were collected from cattle and dogs in Zhanjiang and Jieyang, Guangdong, southern China. Tick species were morphologically identified by an experienced technician and confirmed by sequencing the mitochondrial 16S ribosomal RNA (16S rRNA) gene . The samMeta-transcriptome sequencing was conducted as previously described . In briede novo assemble by the Megahit program, and the resulting contigs were compared against the nr database using the diamond BLASTX program. The confirmed viral contigs with assembly overlaps or from the same scaffold were merged using the SeqMan program . Reads were mapped to the target contigs using Bowtie 2, and the integrated genomics viewer (IGV) was used to check for assembly faults in order to validate the assembly results. Gaps between the contigs were filled by RT-PCR and Sanger sequencing. The genomic terminus of targeting viruses was determined using the 5′/3′ RACE kits . The complete viral genome was confirmed by Sanger sequencing with overlapping primers that covered the entire viral genome.Sequencing reads were adaptor- and quality-trimmed using the FASTP program, followed by The discovered viruses were classified based on the nucleotide (nt) and amino acids (aa) identities. If the species demarcation criteria remain unclear within a genus, a novel viral species is defined if it holds less than 80% nt identity across the complete genome or less than 90% aa identity of the RNA-dependent RNA polymerase (RdRp) domain with known viruses. All the novel viruses were named “Guangdong tick,” followed by common viral names according to their taxonomy. Viruses classified into established taxonomies were marked with “Guangdong (GD)” to distinguish them from other viral strains.Reference virus sequences were acquired from the GenBank database and examined using Mafft v7.450 to determine the phylogenetic relationships of the identified viruses . Phylogen = 306) and dogs (n = 33) in Jieyang (n = 242) and Zhanjiang (n = 97) of Guangdong Province, China, including 243 R. sanguineus and 96 R. microplus  of total viral reads, including 1.04% , 22.89% , and 27.49% reads  mapped to the genomes of LTV, BDTPV1, and BDTPV2, respectively . We obtaR. sanguineus tick pools.The genomes of these viruses contained bi-segments, large (L) and small (S) segments, encoding the putative RdRp and nucleoprotein, respectively . As showUukuvirus group. They were genetically related to those identified in Brazil, Thailand, Colombia, or Trinidad and Tobago  in 3 pools were matched with WTV2  from 3 ith WTV2 .The complete genomes of miviruses were obtained and named MIV GD and WTV2 GD JY02-3 and ZJ2103. Both the WTV2 and MIV genome of the study were confirmed to belong to the circular structure by performing PCR with the forward primer located in 3′ of the genome and the reverse primer in the 5′ part . The genPhylogenetically, both chuviruses identified in this study were grouped into the Mivirus clade and closely related to the viral strains discovered in other countries, such as Brazil, Thailand, Colombia, or Trinidad and Tobago .Metagenomics revealed 3.42% of viral reads  mapped to WTV1 , and fouHepacivirus in the family Flaviviridae (Approximately 6.1% of reads (233/3642) in GDZJ/02 collected from cattle in Zhanjiang were assigned to BovhepV in the genus iviridae . Three siviridae .Metagenomics showed that 56,128 reads  in five pools were annotated to GTQV . We idenGTQV shared 57.4–86.2% aa identity to the quaranjaviruses , whose POrthomyxoviridae. Compared with other quaranjaviruses, the GTQV genome had a specific terminal sequence  were mapped with the reovirus-like virus in the hanjiang . The comhanjiang .Rhipicephalus in Sudan . Alpharicinrhavirus is a genus of Alpharhabdovirinea, containing three species of viruses discovered in ticks. Wuhan tick virus 1 (WTV1), belonging to the Alpharhabdovirinea subfamily, was first detected in R. microplus from Wuhan, China  in the family Orthomyxoviridae, Guangdong tick orbivirus (GTOV) in the family Reoviridae, and Guangdong tick Manly virus (GTMV) of an unclassified family, in which most of them were genetically related to the virus strains in countries outside China, and GTQV, GTOV, and GTMV may represent novel viral species. These findings provided the evidences of the long-distance spread of these TBVs in Guangdong, southern China, suggesting the necessity and importance of TBV surveillance.This study investigated the virome of The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/ZW and QL conceived the project. JM, JL, WH, LZ, and KZ collected the samples. LG, MC, WL, JZ, QJ, HH, and LZ conducted experiment. LG and QL analyzed the data and drafted the manuscript. All authors were involved in critically revised the manuscript and approved the final version.The study was funded by the Science and Technology Innovation Project in Foshan, Guangdong Province, China (2020001000151), the Pearl River Talent Plan in Guangdong Province of China (2019CX01N111), and Guangzhou Science and Technology Project of China (202103000008).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.