Culicoides) are vectors of arboviruses of both veterinary and medical importance. The surge of emerging and reemerging vector-borne diseases and their expansion in geographical areas affected by climate change has increased the importance of understanding their capacity to contribute to novel and emerging infectious diseases. The study of Culicoides virome is the first step in the assessment of this potential. In this study, we analyzed the RNA virome of 10 Culicoides species within the geographical area of Thrace in the southeastern part of Europe, a crossing point between Asia and Europe and important path of various arboviruses, utilizing the Ion Torrent next-generation sequencing (NGS) platform and a custom bioinformatics pipeline based on TRINITY assembler and alignment algorithms. The analysis of the RNA virome of 10 Culicoides species resulted in the identification of the genomic signatures of 14 novel RNA viruses, including three fully assembled viruses and four segmented viruses with at least one segment fully assembled, most of which were significantly divergent from previously identified related viruses from the Solemoviridae, Phasmaviridae, Phenuiviridae, Reoviridae, Chuviridae, Partitiviridae, Orthomyxoviridae, Rhabdoviridae, and Flaviviridae families. Each Culicoides species carried a species-specific set of viruses, some of which are related to viruses from other insect vectors in the same area, contributing to the idea of a virus-carrier web within the ecosystem. The identified viruses not only expand our current knowledge on the virome of Culicoides but also set the basis of the genetic diversity of such viruses in the area of southeastern Europe. Furthermore, our study highlights that such metagenomic approaches should include as many species as possible of the local virus-carrier web that interact and share the virome of a geographical area.Biting midges ( Analyses of arthropod-borne viruses (arboviruses) have spurred the interest of the scientific community. Epidemics caused by yellow fever virus , Zika viCulicoides encompasses the most important virus vectors in the family Ceratopogonidae and consists of more than 1,300 identified species , ticks (Ixodidae), sand flies , black flies (Simuliidae), and biting midges (Ceratopogonidae), the biting midges have received less attention regarding their ability to host or carry viruses, despite the fact that they are the most abundant hematophagous insects worldwide . The gen species  flourish animals . Most CuCulicoides are of both medical and veterinary importance not only because of their numerical abundance and severity of biting activity but because they carry, either actively or mechanically, a wide range of pathogens, including bacteria, viruses, protozoa, and nematodes ], Rhabdoviridae , and Peribunyaviridae family  (ematodes . Culicoirg virus  (SBV) anrg virus , mainly rg virus  (OROV) r (AKAV)] . The geo (AKAV)] . Changes (AKAV)]  have ass (AKAV)]  and SBV  (AKAV)] .Culicoides is the vector specificity and competence. Viruses such as BTV are vectored by multiple species in a specific area  was identified in virome screenings in bats several years before its emergence in the same geographic area . Despiter virome . MetagenScotland , C. imic Senegal , and Culin Japan . On the s virome .Culicoides as most species have distinct ecological and ethological characteristics, and control measures may vary greatly. Furthermore, biological control using the identified viruses against a certain Culicoides species may also be considered a possibility. In the context of this study, a holistic approach is attempted by simultaneously identifying the virome of 10 field-collected Culicoides species present in the geographic area of Thrace, Greece, at the southeastern part of Europe, an important path of various arboviruses between the continents of Asia and Europe. In contrast to previous attempts that analyzed either one species or mixtures of species of a taxon  data due to the stochastic nature of TRINITY assembler program. The results of Ion torrent sequencing and TRINITY assembly are summarized in Culicoides species showcased a wide range of viruses, which corresponded to 9 viral families: Solemoviridae, Phasmaviridae, Phenuiviridae, Reoviridae, Chuviridae, Partitiviridae, Orthomyxoviridae, Rhabdoviridae, Flaviviridae, and one marked as unclassified at the family level  possessed a single virus species each. Notably, Culicoides newsteadi and C. imicola did not yield any viral sequences in the context of this study, despite both being notorious virus vectors. The virome of each Culicoides sample is described analytically per examined species in the following paragraphs.All of the examined C. punctatus harbored a punctatus phasma virus, member of the Phasmaviridae viral family, which was also partially assembled. Members of the family Phasmaviridae typically comprise three diverse genomic segments, namely, the nucleocapsid (segment S), glycoprotein (segment M), and RdRp (segment L) (C. punctatus. The assembled N segment of punctatus phasma virus returned the nucleocapsid protein of Wuhan mosquito virus 1 (YP_009305133.1) as best overall BLASTp hit, whereas the assembled L segment matched the corresponding RdRp of Aedes phasmavirus (QOI91399.1) after a BLASTp search.gment L) . HoweverC. submaritimus belonging to the families Reoviridae and Rhabdoviridae, namely, submaritimus reo-like virus and submaritimus rhabdo-like virus, respectively. Four segments of submaritimus reo-like virus were successfully identified, constituting the RNA-dependent RNA-polymerase (RdRp), the RNA-binding protein (RBP), the major core capsid, and the minor core capsid (Two viruses were detected within e capsid . Only thCulex rhabdovirus (QEM39120.1) as top BLASTp hit.Moreover, submaritimus rhabdo-like virus was retrieved with its nearly complete nucleoprotein (N protein), as well as the partially assembled glycoprotein (G protein) and RdRp (L protein), whereas matrix and phosphoprotein transcripts could not be recovered . The besC. puncticollis species, which carried a puncticollis orthomyxo-like virus, member of the Orthomyxoviridae viral family, consisting of four partially assembled genomic segments, specifically one NP and three discrete polymerases  . The remaining PB1 and PB2 polymerases matched PB1 of Wuhan mosquito virus 3 (AJG39091.1) and PB2 of A. alboannulatus orthomyxo-like virus (ASA47421.1), respectively.Only 1 virus was obtained from B1, PB2) . DespiteC. univittatus showed the species was infected with two different viruses, a univittatus sobemo-like virus and a univittatus rhabdo-like virus, belonging to the families Solemoviridae and Rhabdoviridae, respectively. Solemoviridae and sobemo-like viruses typically consist of two discrete genomic segments, each one encoding two proteins (Vespa velutina RNA virus 3 (ATY36114.1) after a BLASTp search, whereas the second protein product of the same segment showed the highest identity to the RdRp of Atrato sobemo-like virus 1 (QHA33896.1). Nevertheless, the second segment of univittatus sobemo-like virus encoded only one protein noted as capsid. The assembled capsid of univittatus sobemo-like virus was closely related to the putative coat protein of Atrato sobemo-like virus 2 (QHA33889.1) according to BLASTp results. Univittatus rhabdo-like virus was also detected and recovered successfully from C. univittatus. However, it was assembled only partially as most of its genes were missing except for the encoded RdRp (Culex pseudovishnui rhabdo-like virus (BBQ04832.1) via BLASTp.Field-collected samples of proteins . Similarded RdRp . The assC. obsoletus was one of the most potent virus carriers of the examined species, carrying two viruses belonging to Partitiviridae and Rhabdoviridae families and one unclassified virus. Obsoletus partiti-like virus, obsoletus rhabdo-like virus, and an unclassified obsoletus Chaq virus were detected within the field-collected samples of C. obsoletus. Like all partitiviruses, obsoletus partiti-like virus had a bipartite genome, with each segment encoding the RdRp and capsid proteins of the virus, respectively, which were both assembled successfully (Culex rhabdo-like virus (AXQ04770.1), whereas BLASTp of the assembled glycoprotein returned the glycoprotein of Ohlsdorf virus (ATG83561.1) as a top hit. The RdRp of obsoletus rhabdo-like virus BLASTp matched the polymerase-associated protein of Ohlsdorf virus (ATG83562.1). Notably, C. obsoletus carried an unclassified obsoletus Chaq virus, whose closest relative was the corresponding ORF of Chaq-like virus (QMI58124.1) after BLASTp search , glycoprotein (segment M), and RdRp (segment L) proteins (cataneii bunya-like virus yielded the nucleocapsid of Austropotamobius brown spot virus (QCO84581.1) as top BLASTp hit, whereas the assembled glycoprotein BLASTp matched the glycoprotein of Dar es Salaam virus TZ-189 (QDF82061.1). A BLASTp of the RdRp of cataneii bunya-like virus returned the L protein of Blacklegged tick phlebovirus 1 (ANT80544.1) as top hit. A member of the recently described family Chuviridae, cataneii chu-like virus was detected in this study, but only the NP gene of which could be assembled partially . Another member of the family Rhabdoviridae was found within C. cataneii, named cataneii rhabdo-like virus. The glycoprotein and RdRp genes of cataneii rhabdo-like virus were recovered successfully and assembled partially, whereas the rest of the viral genes could not be retrieved (cataneii rhabdo-like virus glycoprotein. The large protein (RdRp) of Riverside virus 1 (AMJ52373.1) was the top BLASTp hit of cataneii rhabdo-like virus RdRp.Similarly, proteins . The assartially . The NP etrieved . The glyC. circumscriptus carried circumscriptus flavi-like virus, which resembled members of the family Flaviviridae. Although circumscriptus flavi-like virus characteristics have not been described fully, six viral genes were recovered successfully and assembled completely from the field-collected samples of C. circumscriptus (circumscriptus flavi-like virus showed the highest identity to the putative capsid protein of Wuhan aphid virus 2 (QDF44112.1), whereas the protein product following capsid transcription BLASTp matched the hypothetical protein of Wuhan aphid virus 1 (BBV14760.1). Similarly, an extra protein product was found before glycoprotein transcription highly similar to the hypothetical protein of Wuhan aphid virus 1 (YP_009179380.1), whereas the main glycoprotein product itself was closer to the putative glycoprotein of Wuhan aphid virus 2 (QDF44110.1) via BLASTp searches. The remaining two assembled genes displayed high similarities with the corresponding NS3 and NS5 transcribed genes of flavi-like viruses. The NS3 of circumscriptus flavi-like virus retrieved NS3-like protein of soybean thrips virus 4 (QPZ88419.1) as the top BLASTp hit. Furthermore, its assembled NS5 showed the highest identity to the NS5-like protein of Wuhan aphid virus 1 (QPZ88370.1).Interestingly, scriptus . NotablyC. haranti also presented interesting findings in the tested field-collected samples as they harbored haranti chuvirus, a member of the recently described family Chuviridae. With little prior knowledge regarding the genomic structure and other characteristics of this viral family, two genes of haranti chuvirus were recovered successfully but assembled only partially  (Sobemovirus identified in the bird Abrornis inornate (Solemoviridae sp. QJI53819.1)   identified in the same region in Anopheles algeriensis mosquitoes (cataneii rhabdo-like virus formed their own cluster between Merhaviruses and Ohlsrhaviruses (Culiseta longiareolata and Coquillettidia richiardii) in the same area  . The onl53819.1) . Punctato, Japan . Interes99886.1) . Anotherdo virus . Submariubfamily . Four Cuectively . Univittsquitoes . Obsoletaviruses . Obsoletame area . Harantie family . Puncticus genus . Circumsus genus . Obsoletus genus . Didymotaq virus .Solemoviridae, Rhabdoviridae, Phasmaviridae, and Partitiviridae families, the novel viruses reported in this study were closely related to viruses previously identified in mosquitoes in the area of Thrace, Greece.In terms of phylogeography, the viruses identified in this study did not cluster according to area, country, or continent. Although in some cases such as in Culicoides were field-collected using CDC light traps with Photo Cell and Air Gate   . Collect States)  during afeatures .Culicoides were homogenized using pellet pestle , and total RNA was extracted by TRIzol reagent  according to the manufacturer’s protocol. As verification of the morphological identification, DNA barcoding was done using standard cytochrome c oxidase subunit 1 (COI) polymerase chain reaction (PCR) and Sanger sequencing. One microgram of the RNA extract was reverse-transcribed at 42°C for 60 min, using M-MLV reverse transcriptase  according to the manufacturer and COI_R primer (5′-AAAAATTTTAATTCCAGTTGGAACAGC-3′). Universal primers COI_F (5′-GGATTTGGAAATTGATTAGTTCCTT-3′) and COI_R were used to amplify a 600-bp PCR product. The PCR reaction mixture contained 0.25 × GC buffer, 1.5 mM MgCl2, 1 mM dNTPs mix, 0.2 μM of each primer, 1.5 U KAPA Taq DNA polymerase , and 1 μL of cDNA. The thermal profile of the PCR included 40 cycles of denaturation at 95°C for 30 s, annealing at 50°C for 45 s, and elongation at 65°C for 1 min, and a final elongation step at 65°C for 7 min. PCR products were purified using the NucleoSpin Gel and PCR Clean-up purification kit . Sanger sequencing was performed on the PCR product, and the resulting sequence was analyzed using the Barcode of Life Data System V4 platform   .Culicoides species, the COI sequences of the respective species  was used for the construction of the phylogenetic trees. In case the RdRp segment was not assembled, nucleocapsid (segment S or N) or glycoprotein (segment M or G) amino acid sequence was used. The length of the input sequence varied, depending on the length of the assembled contigs. The respective virus amino acid sequence obtained from ExPASy Translate online tool  accordinz-direct  by using website  for the  website ; alignme website ; phyloge website , based o website .Culicoides ecology and biology. As climate change alters vector ecology and distribution, vector-borne pathogens find new paths in naive territories , and ticks. Although viruses from mosquitoes seem to populate most of the phylogenetic clusters, the bias resulting from the extensive virome analysis in mosquitoes should be taken into consideration. Viruses similar to puncticollis othomyxo-like virus have been identified before only in mosquito species of the genera Culex   and Coquillettidia (QHA33915.1) (C. longiareolata (QRD99865.1) and C. richiardii (QRD99905.1) in Thrace  and near-complete or complete (6) genomes corresponding to species of the entified –4. The pra Culex  and Aedex (Aedes . Obsoletubgenus) , and Cul) (Culex . The afoke virus  identifi33915.1) . Interes33915.1) , Colombin Thrace . Similar Belgium . This coin China . ComparaCulicoides virome, it is not surprising that only Carapha virus identified in C. arakawae in Japan (punctatus phasma virus)  . To be nhe genus . Often, he genus . Hypothehe genus  or contahe genus . Studieshe genus . Submarieviously .Chuviridae family were identified in three Culicoides species, namely, obsoletus Chaq virus, haranti chuvirus, and cataneii chu-like virus. Chaq viruses were proposed to be satellite viruses of other viruses, as in its initial identification, Chaq virus contigs were always present together with Galbut virus during viral metagenomics (Chuviridae is a family of arthropod viruses often found only in metagenomic studies with variable genomic structure (cataneii chu-like virus with Wuchang cockroach virus 3 within the phylogenetic tree and their distance from Tacheng tick virus 5 and Bole tick virus 3 favored a segmented genome structure depiction (type II) as proposed by Flaviviridae have been shown to form EVEs , cataneii bunya-like virus was closer to a group of viruses from aphids  can be found below: IK designed the study. IK and SV obtained funding for the project. AN and PP conducted fieldwork and collected samples. MC performed morphological species identification. KK, MB, and EG performed experiments. KK and ND performed the analysis. KK, MB, and IK wrote the manuscript with input from all authors. All authors contributed to the article and approved the submitted version.PP was employed by the company Evrofarma S.A. AN was employed by the company Geotechno Ygeionomiki O.E. The remaining 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.


First pass reperfusion (FPR), that is, excellent reperfusion  2C-3) in one pass, after endovascular treatment (EVT) of an occluded artery in the anterior circulation, is associated with favorable clinical outcome, even when compared with multiple pass excellent reperfusion (MPR). In patients with posterior circulation ischemic stroke (PCS), the same association is expected, but currently unknown. We aimed to assess characteristics associated with FPR and the influence of FPR versus MPR on outcomes in patients with PCS.We used data from the MR CLEAN Registry, a prospective observational study. The effect of FPR on 24-hour National Institutes of Health Stroke Scale (NIHSS) score, as percentage reduction, and on modified Rankin Scale (mRS) scores at 3 months, was tested with linear and ordinal logistic regression models.Of 224 patients with PCS, 45 patients had FPR, 47 had MPR, and 90 had no excellent reperfusion (eTICI <2C). We did not find an association between any of the patient, imaging, or treatment characteristics and FPR. FPR was associated with better NIHSS (−45% (95% CI: −65% to −12%)) and better mRS scores (adjusted common odds ratio (acOR): 2.16 (95% CI: 1.23 to 3.79)) compared with no FPR. Outcomes after FPR were also more favorable compared with MPR, but the effect was smaller and not statistically significant (NIHSS: −14% (95% CI: −51% to 49%), mRS acOR: 1.50 (95% CI: 0.75 to 3.00)).FPR in patients with PCS is associated with favorable clinical outcome in comparison with no FPR. In comparison with MPR, the effect of FPR was no longer statistically significant. Nevertheless, our data support the notion that FPR should be the treatment target to pursue in every patient treated with EVT. Excellent reperfusion  2C-3) in one pass, first pass reperfusion (FPR), after endovascular treatment (EVT) of an occluded artery in the anterior circulation, is associated with favorable clinical outcome.No studies have been published about the association between FPR and outcomes in patients with PCS. In patients with anterior circulation ischemic stroke, FPR is associated with favorable outcome, independently of patient, imaging, and treatment characteristics, even when compared with multiple pass excellent reperfusion (MPR).We used data from the Multicenter Randomized Clinical Trial of Endovascular Treatment for Acute Ischemic Stroke in the Netherlands (MR CLEAN) Registry. This was a prospective observational study in all 18 centers performing EVT in the Netherlands. All patients undergoing EVT for acute ischemic stroke in the anterior and posterior circulation were registered, except for those who were treated in the BASICS trial.From the MR CLEAN Registry, we included patients aged 18 years or older, who had a symptomatic occlusion of the vertebral, basilar, or posterior cerebral artery confirmed by baseline computed tomography angiography. These data concerned patients who were treated with EVT between March 16, 2014 and December 31, 2018.An imaging core laboratory analyzed all patient imaging. The members of the core laboratory were blinded to all clinical data. Reperfusion grade was measured according to the eTICI scale on final digital subtraction angiography (DSA) by the core laboratory. The number of attempts used to achieve reperfusion was based on the information given by the local treating interventionalist.FPR was defined as a single pass of the device, without rescue treatment with intra-arterial thrombolytics, resulting in complete or near-complete reperfusion of the large vessel occlusion and its downstream territory: eTICI 2C-3. MPR was defined as eTICI 2C-3 after more than one pass or after one pass followed by rescue treatment with intra-arterial thrombolytics. No excellent reperfusion (NER) was defined as eTICI <2C independently of the number of passes.Patient characteristics included: age, sex, history of atrial fibrillation, history of hypertension, history of diabetes mellitus, history of myocardial infarction, history of peripheral artery disease, history of stroke, history of hyperlipidemia, smoking, use of antiplatelet agents, use of vitamin K antagonists, use of direct oral anticoagulants, National Institutes of Health Stroke Scale (NIHSS) score at baseline, and pre-stroke modified Rankin Scale (mRS) score.Imaging characteristics (scored by the core laboratory) included: level of obstruction, hyperdense artery sign, posterior circulation collateral score,Treatment characteristics included: estimated time of large vessel occlusion (eLVO) to presentation at intervention hospital, presentation at intervention hospital to groin time, intravenous alteplase treatment, general anesthesia, and used device.In patients with transient or mild neurological symptoms with secondary worsening consistent with the large vessel occlusion, the moment of secondary worsening was considered as the eLVO.The percentage change in 24-hour NIHSS (±12 hours) was used as the primary outcome. This has been shown to be more closely related to EVT and reperfusion than the mRS score at 3 months, and has a good predictive value for long-term stroke outcome.All baseline data were reported as crude. If successful reperfusion was not achieved during EVT, we used the time of last contrast bolus injection as the final reperfusion time. For the use in regression models we imputed missing data using multiple imputation with R  based on relevant covariates and outcomes. Any mRS score of 0 to 5 at follow-up assessed within 30 days of symptom onset was considered invalid and treated as missing.We compared baseline characteristics of patients with FPR with patients without FPR using descriptive statistics. To investigate the association between patient, imaging, and treatment characteristics and FPR we used an univariable logistic regression model and selected all variables with a P value ≤0.2. These selected variables were used in a multivariable logistic regression model with a backward stepwise selection procedure with three steps. In each additional step, variables with a P value >0.2 were dropped, except for age and sex which were forced into the model. In step one, we tested all patient characteristics. In step two, we added all imaging characteristics to the remaining variables from step one. In step three, we added treatment characteristics to the remaining variables from step two. The final model consisted of all variables with a P value ≤0.2 and age and sex.We analyzed the association between FPR and outcomes, adjusted for characteristics associated with FPR. First, we compared outcomes between FPR and no FPR. Second, we compared FPR with MPR. We used a linear regression model to analyze the NIHSS score at 24 hours and presented percentage change with 95% confidence intervals (CIs). Patients who had died before the time point of NIHSS assessment was reached, received the maximum NIHSS score of 42. The NIHSS was then log10 transformed, to better meet the assumption of normally distributed residuals in linear regression.Of all the patients in the MR CLEAN Registry, 264 adult patients had PCS. We excluded 40 patients who did not receive mechanical thrombectomy, because arterial access to the intracranial vasculature was not achieved or first DSA before EVT showed (spontaneous) reperfusion. Therefore, 224 patients were included . FPR wasWe analyzed the association of patient, imaging, and treatment characteristics with FPR compared with no FPR (MPR+NER). In the backward stepwise selection procedure only a history of hyperlipidemia was selected, next to age and sex which were forced into the model. None of these variables were associated with FPR (history of hyperlipidemia adjusted odds ratio (aOR) 1.17 (95% CI: 0.99 to 1.39)). We also analyzed the association of patient, imaging, and treatment characteristics with FPR contrasted with MPR. We selected age, sex, history of hyperlipidemia, and baseline pc-ASPECTS from the backward stepwise selection procedure. However, none of these variables were associated with FPR (history of hyperlipidemia aOR 1.27 (95% CI: 0.98 to 1.63), pc-ASPECTS aOR 1.04 (95% CI: 0.95 to 1.13)).In the univariable regression analysis, FPR compared with no FPR led to a statistically significant decrease in 24-hour NIHSS score of −45% (95% CI: −64 to −14) . Compare10.1136/neurintsurg-2021-017507.supp1Supplementary dataProcedure time for FPR was shorter than for no FPR . When prIn the univariable regression analyses, FPR led to more favorable mRS scores at 3 months compared with no FPR (cOR 1.96 (95% CI: 1.14 to 3.39)) and compared with MPR (cOR 1.33 (95% CI: 0.69 to 2.53)) . The shiIn our study, FPR was associated with favorable clinical and functional outcomes. There were no patient, imaging, or treatment characteristics associated with FPR. In a study of patients with PCS treated with aspiration first, a history of diabetes, onset-to-groin time, and cardioembolic etiology were found as predictors of FPR (in this study defined as eTICI ≥2B in one attempt).The association between FPR and 24-hour NIHSS is comparable to our results in patients with anterior circulation ischemic stroke .The effect of FPR over MPR and NER can be partly explained by a shorter procedure time. We showed that the effect of FPR on outcomes was not completely reduced when we adjusted for procedure time. A higher maneuver count is associated with a poor outcome despite successful recanalization.Parallel to the data collection of the MR CLEAN Registry, the BASICS trial was performed. Of all the hospitals in the MR CLEAN Registry, 10 participated in the BASICS trial and eight did not participate in the BASICS trial. There could be some selection bias in our cohort because we only collected data of patients treated with EVT outside the BASICS trial. In separate analyses of our data there were no differences in patient characteristics and outcomes between patients treated in a BASICS trial center and patients treated in an non-BASICS trial center.All our imaging was assessed by an independent core laboratory. We used very strict definitions for these assessments, and when the quality of imaging was low or there were only a few images available the imaging variables were scored as missing. Registries in general are prone to missing values. However, all data were verified by our study coordinators, and we made every effort to retrieve the missing data.During the inclusion period of the MR CLEAN Registry there was no clear evidence for a benefit of EVT over best medical management on outcome in patients with a basilar artery occlusion.Another limitation is the assessment of the eTICI score, which is less reliable in the posterior circulation. The assessment is more difficult due to the interference with abundant collateral flows, incomplete visualization of the perforating arteries to the brain stem, and the necessity to consider the antegrade flow from the anterior circulation.FPR in patients with PCS is associated with favorable clinical outcome in comparison with no FPR. In comparison with MPR, the effect of FPR was no longer statistically significant. Nevertheless, our data support the notion that FPR should be the treatment target to pursue in every patient treated with EVT.


SLC26A9 belongs to the solute carrier family 26 (SLC26), which comprises membrane proteins involved in ion transport mechanisms. On the basis of different preliminary findings, including the phenotype of SlC26A9-deficient mice and its possible role as a gene modifier of the human phenotype and treatment response, SLC26A9 has emerged as one of the most interesting alternative targets for the treatment of cystic fibrosis (CF). However, despite relevant clues, some open issues and controversies remain. The lack of specific pharmacological modulators, the elusive expression reported in the airways, and its complex relationships with CFTR and the CF phenotype prevent us from conclusively understanding the contribution of SLC26A9 in human lung physiology and its real potential as a therapeutic target in CF. In this review, we summarized the various studies dealing with SLC26A9 expression, molecular structure, and function as an anion channel or transporter; its interaction and functional relationships with CFTR; and its role as a gene modifier and tried to reconcile them in order to highlight the current understanding and the gap in knowledge regarding the contribution of SLC26A9 to human lung physiology and CF disease and treatment. SLC26A9 belongs to the solute carrier family 26 (SLC26), which comprises membrane proteins related to the phylogenetically older SLC26-SulP gene family. As part of the APC superfamily, such proteins operate as electroneutral or electrogenic exchangers of monovalent and divalent anions and as anion channels . The mamIn this context, SLC26A9 is under investigation as one of the most interesting targets. Such interest arises from various preliminary observations: (1) it was originally described as a protein expressed in the lung ; (2) givIn this review, we summarized the various results dealing with SLC26A9 expression at the tissue, cellular, and subcellular levels; its structure and function as anion channel or transporter; its interaction and functional relationships with CFTR; and its role as gene modifier, and tried to reconcile them in order to highlight the current understanding and the gap in knowledge regarding the contribution of SLC26A9 to human lung physiology and CF disease and treatment. The original description of SLC26A9 as a lung-expressed protein drew attention to SLC26A9 regarding its possible role in airway physiology and disease . HoweverAs mentioned above, the first observation by Lohi and colleagues detected SLC26A9 by immunohistochemistry with a self-produced antibody , the prostate, and the stomach (fundus/corpus and antrum region) as the main sites of expression, whereas lower levels have been found in the proximal duodenum  18,25,2,225,26. Given its possible role in CF, SLC26A9 expression has been investigated in epithelial cells collected from CF and non-CF individuals and cultured under air–liquid conditions. Different research teams have provided evidence of some SLC26A9 expression at the mRNA level ,28,29,30−- and BHK-transfected cells, particularly if cultured on porous supports [Concerning the subcellular localization, additional and sometimes controversial data come from studies relying on the heterologous expression of SLC26A9 in Xenopus oocytes and various immortalized cell lines. The transfection of the full-length coding sequence in HEK-293 cells resulted in a high rate of protein expression with the accumulation of intracellular aggregates and almost an absence of SLC26A9 protein at the plasma membrane ,33. Retesupports ,35,36. Isupports .− and HEK-293 cells was observed only by transfecting SLC26A9 constructs carrying a modification of the C-terminal coding sequence, including the deletion of the PDZ or/and the STAS domains  folding such as UraA and NBCe1 . Indeed,SLC26A9 exists as a homodimer. Each protomer can be divided into a TM domain and a cytosolic domain composed, respectively, of 14 transmembrane segments and the N- and C-terminal tails of the protein. As modeled in The C-terminal sequence of SLC26A9 displays unique features and appears folded in the intracellular pocket lined by TM5, TM8, TM10, and TM12. Such a position alters the surface charge from positive to negative. In this way, the C-terminal may reduce the ion accessibility from the cytosolic side, possibly modulating the ion transport kinetics . AccordiMoreover, a similar gain of function was obtained by mutating residues within the sequence involved in the interaction between the C-terminus and the TM12-TM5, further highlighting an inhibitory feature of the SLC26A9 C-terminus.− and one Na+, were resolved in the high-resolution structure provided by Chi and colleagues [− and Na+ to the substrate-binding site, a behavior corresponding to the rapid Cl− flux observed in SLC26A9, and proposed Cl− as being the main ion transported [Multiple ions, possibly two Cllleagues . The molnsported ,33.−, and C₂O₄²− by isotope uptake assays in Xenopus oocytes expressing the human protein [−/HCO3− exchange and Cl−-independent HCO3− extrusion [−/HCO3− exchange, electrogenic Na+/nAnion− cotransport, and anion channel [− secretion with linear current–voltage relationship [The transport properties of SLC26A9 have been investigated by means of various experimental approaches. SLC26A9 was found to be permeable to SO₄²-, Cl protein . Intracextrusion . Subsequ channel . The anitionship ,34,37. Rtionship ,33.Several studies have investigated the physical and functional interaction between SLC26A9 and CFTR.Xenopus oocytes and resulted in the inhibition of SLC26A9 activity [A possible direct interaction between SLC26A9 and CFTR, involving the STAS domain of SLC26A9 and the regulatory domain of CFTR, was depicted in activity . Many ot− and 16HBE14o−), SLC26A9 overexpression and silencing resulted in increased and reduced CFTR expression and function, respectively [Concerning CFTR activity, the co-transfection of SLC26A9 and wt-CFTR in HEK-293 cells resulted in enhanced forskolin-stimulated CFTR-dependent current . Importaectively .Xenopus laevis oocytes) [Concerning SLC26A9 activity, some studies reported a low contribution to anion transport, either in the presence or in the absence of CFTR ,42. Rathoocytes) ,35,36. SSuch relationships were further confirmed by studies involving CFTR modulators. In BHK cells transfected with F508del-CFTR and SLC26A9, the treatment with low temperature or the CFTR corrector VX-809  increased the SLC26A9 cell surface expression . Similar−-free condition and was inhibited by GlyH-101, niflumic acid, and DIDS , but not by a specific CFTR inhibitor, CFTRinh-172 [The contribution of SLC26A9 to transepithelial anion secretion has been investigated in different epithelial models. The heterologous expression of SLC26A9 in FRT cells cultured on porous supports increased the baseline anion current measured by short-circuit current recordings . Such acRinh-172 ,34. A coRinh-172 ,35. ThisRinh-172 ,35. Thes− secretion in the airways of wild-type but not SLC26A9-deficient mice, with the latter thus exhibiting airway mucus obstruction [In mice airways, SLC26A9 may play a clearer role, particularly under inflammatory conditions characterized by a Th-2 response. Indeed, IL-13 treatment increased Cltruction . SLC26A9truction .Different studies have defined SLC26A9 as a modifier gene of the gastrointestinal tract manifestations of cystic fibrosis. Various SNPs at its 5′ have been found to be associated with meconium ileus, pancreatic damage, immunoreactive trypsinogen at birth, and CF-related diabetes (CFRD) ,45,46. AP. aeruginosa and residual lung function, SLC26A9 was shown not to be involved in lung function [Nevertheless, the association of SLC26A9 SNPs with lung function is controversial. Several SLC26A9 SNPs have been investigated. For example, the rs7512462 SNP (T > C) occurring in intron 5 is highly debated. The minor allele C was associated with improved lung function in individuals with at least one G551D allele  . In anotfunction .Additionally, the association between SLC26A9 SNPs and the therapeutic efficacy of CFTR modulators has been the focus of different investigations. The rs7512462 SNP explained 22% to 28% of the response variability to ivacaftor treatment for patients carrying G551D mutation, with improved response associated with the C allele ,48. MoreSLC26A9 is emerging as a focus for the development of novel therapeutic strategies for CF disease. Already-approved drugs, based on potentiator and correctors able to rescue the function of mutant CFTR, are showing very promising results in the treatment of several CF patients, particularly those carrying gating mutations and F508del. However, there is a significant fraction of CF patients with “undruggable” mutations. In such cases, as well as for possible adjuvant or synergistic approaches, a strategy based on an alternative target can be considered. In the case of SLC26A9, it could be speculated that the potentiation of its anion transport activity may circumvent the reduced chloride secretion due to the loss of function of CFTR .However, the knowledge of the role of SLC26A9 in human physiology and CF disease is still in its infancy. Besides different studies seem to indicate a relevant role of this protein in the gastrointestinal tract, the contribution of SLC26A9 to the lung function is still elusive, highlighting the need for further investigations. For example, despite a few studies reporting some SLC26A9 expression in human bronchial airways ,24, othe


Cell surface receptors play a key role in a virus’ ability to recognize and invade cells and tissues, which basically defines viral pathogenicity. The ongoing COVID-19 pandemic caused by the SARS-CoV-2 virus has shown us the central role of the receptor(s) in the infectious entry of pathogenic viruses. The importance of receptors is further emphasized by their role as vaccine targets. However, as we have been witnessing in the case of SARS-CoV-2, viral surface proteins may rapidly mutate during virus evolution, which may make viruses more infectious and increase the speed by which viruses are able to infect cells. Conversely, viruses may also adapt to avoid neutralizing antibodies, alleviating the effects of vaccines. Viruses has managed to attract four original articles and two reviews on a wide range of topics. Papers about RSV receptor interactions, airway epithelial cell tropism, or structural comparisons between the receptor binding of pathogenic viruses are included in the issue and have attracted a lot of interest, with more than 1000 views per article—these numbers are still increasing. The various natures of the papers in addition to the viruses studied indicate that there is still room for receptor studies; we are far from understanding virus receptor–cell surface protein interactions and, more importantly, how they could be used for the benefit of humankind, e.g., in antiviral and vaccine development. I express my sincere thanks to all authors who contributed to this Special Issue, “Virus receptors and viral tropism”.While COVID-19 has severely affected research on topics other than the SARS-CoV-2 virus, this Special Issue, “Virus receptors and viral tropism”, of


Orolabial and genital herpes are common conditions caused by herpes simplex virus (HSV) that affect many individuals. Although skin findings may mimic other conditions, an HSV infection may still be diagnosed clinically. We present a case of polymerase chain reaction (PCR) negative orolabial herpes in a young male with a high-risk social history. Although testing was negative, oral HSV was clinically diagnosed and the patient was adequately treated with IV acyclovir. Both clinical history and physical examination are essential to provide an accurate diagnosis even in cases presenting with a negative diagnostic test result. Herpes simplex virus (HSV)-1 and HSV-2 are responsible for primary and recurrent orolabial and genital herpes, respectively. Although infrequent, HSV-2 also has the potential to cause orolabial herpes. Clinically, orolabial herpes presents as superficial ulcerative lesions of the tongue and surrounding oral mucosa [A 33-year-old male with a history of genital herpes and IV drug abuse presented with a painfully swollen tongue of two-day duration. He began oral acyclovir four days prior to presentation for recurrent genital herpes. Within two days of treatment, he experienced a “tingling” sensation of the tongue, leading to pain and edema. He had unprotected sexual intercourse and receptive oral sex with multiple male partners in the past month. Physical exam revealed large hemorrhagic erosions with scalloped borders along the lateral surface of the tongue. The large white lesion was not detachable upon scraping. The lower lip demonstrated erosions with overlying slough and yellow crust Figures -2. AdditFollowing primary infection of HSV-1 or HSV-2, the virus lies dormant in the nervous system  causing recurrent breakouts, which can be frustrating for patients. Reactivation occurs in response to a trigger stimulus such as psychological stress, fatigue, sunlight, exposure to cold or heat, sexual intercourse, and immunosuppression . ReactivDuring the initial work-up, our differential diagnosis included Behçet’s syndrome, aphthous stomatitis, erythema multiforme, and Stevens-Johnson syndrome (SJS) as they all present with oral and/or genital lesions. Aphthous ulcers do not involve the keratinized epithelium, such as the lower lip erosions seen in our patient, and Behçet’s syndrome is a multisystem inflammatory disease that also has ocular, neurological, and vascular manifestations. While oral ulcers are frequently the presenting sign of Behçet’s syndrome, our patient’s history of unprotected oral intercourse and rapid response to acyclovir is consistent with herpes infection . ErythemClinicians should always have a high index of suspicion for HSV in high-risk patients who present with similar signs and symptoms. Multiple tests may have to be completed just to yield accurate results . StudiesHSV is a prevalent virus with a relapsing and chronic course that affects a large portion of our society. This burdensome virus is known for its pathognomonic clinical presentation of numerous clear, fluid-filled vesicles upon an erythematous base. However, it is uncommon for HSV to primarily present on the tongue, straying from its common presentation on the lips. If testing is completed and a negative PCR result is obtained, this may complicate the picture, further leading clinicians down a path of misdiagnosis with delayed treatment. A careful history and physical examination are vital for early symptomatic management and to exclude clinical mimickers from the differential diagnosis.


Varroa destructor, is associated with high levels of colony losses. Divergent DWV types may differ in their pathogenicity and are reported to exhibit superinfection exclusion upon sequential infections, an inevitability in a Varroa-infested colony. We used a reverse genetic approach to investigate competition and interactions between genetically distinct or related virus strains, analysing viral load over time, tissue distribution with reporter gene-expressing viruses and recombination between virus variants. Transient competition occurred irrespective of the order of virus acquisition, indicating no directionality or dominance. Over longer periods, the ability to compete with a pre-existing infection correlated with the genetic divergence of the inoculae. Genetic recombination was observed throughout the DWV genome with recombinants accounting for ~2% of the population as determined by deep sequencing. We propose that superinfection exclusion, if it occurs at all, is a consequence of a cross-reactive RNAi response to the viruses involved, explaining the lack of dominance of one virus type over another. A better understanding of the consequences of dual- and superinfection will inform development of cross-protective honey bee vaccines and landscape-scale DWV transmission and evolution.Deformed wing virus (DWV) is the most important globally distributed pathogen of honey bees and, when vectored by the ectoparasite  Apis mellifera) are globally important pollinators of wild flowers and agricultural crops, and the source of honey, with annual global production worth in excess of $7bn ..S and VVE replicated extensively, albeit somewhat delayed when compared with VDDE-only infected pupae, in pupae that had received VVV as the primary virus . In arthropods antiviral RNAi response acts via generation of short double stranded RNAs (siRNA) from virus RNA replication intermediates through cleavage by the enzyme Dicer. These are further used by the RNA induced silencing complex to target the destruction of complementary sequences , 60. HenSupplementary materials


Reported for the first time in 1955 in Malaysia, Tembusu virus (TMUV) remained, for a long time, in the shadow of flaviviruses with human health importance such as dengue virus or Japanese encephalitis virus. However, since 2010 and the first large epidemic in duck farms in China, the threat of its emergence on a large scale in Asia or even its spillover into the human population is becoming more and more significant. This review aims to report current knowledge on TMUV from viral particle organization to the development of specific vaccines and therapeutics, with a particular focus on host-virus interactions. Culex tritaeniorhynchus mosquitoes and then occasionally reported in different surveys in Southeast Asia (SEA) during the 1970s [Tembusu virus (TMUV) is an emerging arbovirus (arthropod-borne virus), which was initially identified in Malaysia in 1955 from he 1970s . As withhe 1970s ,4,5. HowTMUV causes encephalitis and neurological disorders in avian populations, resulting in morbidity rates of up to 90%, associated with damage to the female reproductive system leading to severe loss of egg production in farms . Since 2To date, knowledge on TMUV ecology and biology remains incomplete, and the driving cause of its sporadic emergence remains unknown. This strongly contrasts with other emerging arboviruses causing human diseases such as yellow fever (YFV), DENV, West Nile (WNV), JEV and Chikungunya (CHIKV) viruses. In the last decade, important progress in the understanding of TMUV biology has been made through both experimental animal models and in vitro studies. Here, we review the epidemiology, virus-host interaction and cell biology of avian TMUV infection. We also discuss approaches to increase the knowledge of this virus in order to properly evaluate the associated risk of spillover in humans.As with other flaviviruses, viruses from the TMUV group have a ~11 kb positive strand RNA genome, which is composed of a single open reading frame (ORF) flanked by 5’terminal and 3’terminal non-coding regions (UTR) . The 5’ Despite a lack of information on TMUV replication, this process is thought to be more or less identical to that of other members of the Flavivirus genus . Prior tA range of endocytic pathways are usually used by flaviviruses to penetrate into the cell . Zhang eTo complete its cellular cycle, TMUV has to (1) translate its genomic RNA into viral proteins, (2) replicate its viral RNA genome, (3) encapsidate its genome, (4) assemble the genome into an immature virion in the endoplasmic reticulum, (5) mature the virus particles in the Golgi apparatus and (6) be secreted as infective virus particles in the extracellular space by exocytosis. Although these steps are not well known for TMUV, they do not vary among flaviviruses and have been extensively described elsewhere ,15. TMUVTMUV presents a typical flavivirus structured 3’UTR, which plays an important role in the virus’s fitness . During Flaviviridae family, genus Flavivirus [Aedes- and Culex-transmitted viruses. Aedes-borne viruses, including DENV, YFV and Zika virus (ZIKV), are generally associated with hemorrhagic fevers and non-human primate ancestors, and those transmitted by the Culex genus are associated with meningoencephalitis diseases and bird reservoirs such as JEV, WNV and TMUV [Culex-transmitted MBF are classified depending on the serology in two groups, the Japanese encephalitis serocomplex including JEV and WNV, and the Ntaya serocomplex including Ntaya virus and TMUV.The TMUV members belong to the avivirus . Based oavivirus . Mosquitand TMUV . Culex-tCx. tritaeniorhynchus in Malaysia in 1955 [Cx. annulus and Cx. tritaeniorhynchus mosquitoes was grouped in the cluster TMUV, despite the fact that no TMUV has ever been reported in Taiwan [The TMUV group is genetically distinct within the Ntaya serocomplex and includes homologous strains that were initially considered as single species, including Sitiawan virus , BYD vir in 1955  and repr in 1955 ,25. Fromn Taiwan . Long-din Taiwan ,28. A siSince the beginning of the 21st century, TMUV circulation has been reported in SEA and China . HoweverAltogether, phylogenetic analysis suggests a circulation through insular and land territories of SEA up to the 2000s. Since 2010, TMUV has been circulating actively in China and in SEA, with potential different introductions in China from SEA. The isolation in Taiwan of a strain that groups with the original TMUV cluster has led to speculation that the viruses were introduced by migratory birds, and this may hold true for earlier introductions in Asia. In Cluster 2 and Cluster 3, the phylogenetic association between strains from China and Thailand, and in Cluster 1 between strains from Thailand and Malaysia, indicates a spatial segregation of the TMUV genotypes within Asia. However, despite increasing information on the regional phylodynamic of TMUV, many questions remain about its circulation, particularly about the dynamic of the virus in other Asian countries. Regarding the importance of regional exchange for farming and agriculture trading in Asia, and also with regard to the migration flows of birds, TMUV can be expected to continue its expansion, and this should be investigated.Culex tritaeniorhynchus mosquito captured in Kuala Lumpur, Malaysia, in 1955 and has subsequently been described in different Culex species , with its natural maintenance cycle involving birds and mosquitoes . The etiIt is now clearly established that TMUV is a neurotropic virus for birds, although the mechanisms of diffusion at the neuronal level are still poorly understood. However, a recent study on a duckling model shed light on the modalities of passage from the blood-brain barrier (BBB) to the central nervous system . Thus, iWhile symptoms due to TMUV infection are remarkably well described in birds, disease presentations in other vertebrate hosts are sparse. In 2020, Yurayart et al. reported neuropathogenesis and global dissemination in mice intracerebrally infected with TMUV. The animals exhibited a wide range of clinical signs with additional severe internal organ lesions that led to death . Tang etSince the first isolation of TMUV, several techniques have been developed to study the virus or detect it for diagnostic purposes. The development of diagnostic tools was particularly motivated to prevent large outbreaks in farms and preserve the poultry industry from economic losses. Prevention of outbreaks in countries producing and exporting a large amount of poultry is a major issue, especially for China as it was the first producer of ducks in the world. Kono et al. identified viral particles by electron microscopy in the transformed chicken B lymphocyte cell line BK3 (LSCC-BK3) infected with TMUV . AlthougNowadays, common methods for diagnosis of TMUV include serological detection by enzyme-linked immunosorbent assay (ELISA), based on NS1 protein detection, and the plaque reduction neutralization test (PRNT) ,82. The To ensure its transmission, TMUV must replicate in mosquito vectors prior to a vertebrate host . In paraDifferent types of avian cells have been used to evaluate TMUV biology and include BK3 cells, a transformed chicken B lymphocyte cell line, the chicken macrophage-derived cell line HD-11 and the DF1 cell line, isolated from chicken embryo fibroblasts. Besides specific usage for research, chicken embryo fibroblasts and duck and chicken eggs are routinely used for virus isolation and maintenance ,30,93,94TMUV is also able to replicate in a wide range of non-avian cells which are used to assess the impact of TMUV in mammalian hosts. The primary cells and cell lines used in TMUV investigation are summarized in Flaviviruses trigger a strong innate immune response, which induces the production of a broad range of complementary antiviral molecules. Innate immunity is initiated by recognition of specific viral components, named pathogen-associated molecular patterns (PAMPs), via cellular pattern recognition receptors (PRRs) and was largely investigated in mammals and in birds ,102,103.Several studies investigated the innate immune response against TMUV both in vivo and in vitro in avian and mammalian species. After experimental infection of female ducks or ducklings, Li et al. and Zhang et al. showed a rapid multiplication of the virus in different organs  associated with severe lesions. The infection triggers activation of the RLR and TLR pathways and overexpression of PRRs such as RIG-I or MDA-5. This activation leads to the upregulation of interferon-stimulated genes (ISGs) from the Mx and OAS families in the brain and spleen ,106. UprIn geese, TMUV infection triggers an immune response with an increase in proinflammatory cytokines and interferons in several organs as well as in PMBCs. Interestingly, immunohistochemical analysis presented co-localization of CD8+T cells and TMUV, associated with high cytokine expression, suggesting an activation cascade due to viral infection and leading to the establishment of an antiviral status . MoleculFlaviviruses have developed different immune evasion strategies . Viral pAutophagy is a cellular mechanism of recycling and degradation of cytoplasmic components to prevent cell death and promote homeostasis of cells exposed to an extrinsic or intrinsic stress, including viral infection . DespiteThe endoplasmic reticulum (ER) is an important intracellular organelle in the protein synthesis pathways where proteins are folded and matured before intracellular or extracellular release. The replication cycle of TMUV, as with other flaviviruses, mainly takes place in close interaction with the ER of the infected cell . ER actsAs with some other flavivirus infections, there is no specific treatment for TMUV. The antiviral efficacy of different molecules is currently being evaluated. For example, minocycline, a tetracycline analogue, has shown some efficiency in vitro against neuronal cell death infected by neurotropic viruses. This neuroprotective effect was also found with duck neuronal cells infected with TMUV . EpigallCurrently, there is no commercial vaccine against TMUV. However, regarding the large spread of the virus in Asia since the first outbreak in China in 2010 and the association with a high economic impact on the poultry industry, the development of an effective vaccine is a primary objective. Since the first report of the virus, different research teams have taken on the challenge. In China, attenuated viruses by passaging in embryonated chicken eggs were developed as a live vaccine strategy ,135. TheTwo recombinant duck enteritis viruses were also designed to express a part of the TMUV E protein and the pre-membrane protein. Immunization of ducks led to the production of protective neutralizing antibodies, and further TMUV-challenged animals exhibited resistance to the infection . A reverRecently, vaccine development efforts were dedicated to providing a recombinant vaccine against TMUV. Several studies reported the development of antibodies targeting immunogenic domain regions of the virus particles, particularly the loop of domain II of the E protein . Among tStrategies using attenuated pathogens are widely used for vaccine development. However, an interesting way to develop new vaccines would be to use the strategy based on messenger RNA (RNA vaccine) as it is currently being utilized with success against SARS-CoV2. This type of strategy, coupled with liposome delivery, is already being used in attempts to develop a vaccine against other flaviviruses such as DENV and ZIKV . DespiteAs with other epornitic arboviruses, TMUV has taken advantage of increasingly favorable conditions over recent decades to spread across Asia and invade new territories. The first outbreaks in China in 2010 led to a reconsideration of the danger of this virus. Its transmission is largely facilitated by the conditions of intensive poultry rearing in farms, containing several thousands of animals. In such conditions, TMUV even seems to spread between animals without the intervention of mosquito vectors. Finally, the impact of migratory birds has not been clearly demonstrated but could play a significant role in the expansion of the virus across Asia. Thus, intensive animal husbandry, increasing trade and environmental changes have facilitated the wide spread of TMUV. This situation is all the more worrying since the passage of this virus to humans has been demonstrated and thus could lead to the emergence of a new zoonotic disease. The clinical presentation of TMUV infections in birds is similar to that of JEV, WNV or ZIKV, and it is easy to imagine the impact on the human population if TMUV were to develop therein. Due to the lack of widespread surveillance, the virus has, thus far, been detected in only three Asian countries and actively studied only in China and Thailand. However, the remoteness of these countries and the phylogenetic pattern of the virus suggest that the virus is more widely established in Asia and SEA.In recent years, research on host-pathogen interactions has mainly been focused on avian models, but a better understanding of the biology of the virus in mammals and, in particular, in humans is important. Finally, the presence of potential vectors in non-Asian regions may facilitate spreading outside of Asia as it has recently been the case for other arboviruses. Thus, in order to respond to the challenges posed by this emerging virus, an integrated One Health approach could allow a broader understanding of the different parameters involved in the ecology of TMUV.


Over the last fewyears, a growing interest has surfaced aboutthe possibility of enhancing solar harvester efficiency by couplingphotovoltaic (PV) cells with thermoelectric generators (TEGs). Tobe effective solutions, hybrid thermoelectric-photovoltaic (HTEPV)solar harvesters must not only increase the solar conversion efficiencybut should also be economically competitive. The aim of this paperis to estimate the profitability of HTEPV solar harvesters with noreference to specific materials, relating it instead to their physicalproperties only and thus providing a tool to address research efforttoward classes of HTEPV systems able to compete with current PV technologies.An economic convenience index is defined and used to assess the economicsustainability of hybridization. It is found that, although hybridizationoften leads to enhanced solar power conversion, power costs (USD/W)may not always justify HTEPV deployment at the current stage of technology.An analysis of the cost structure shows that profitability requireslargely enhanced thermoelectric stages, concentrated solar cells,or PV materials with favorable temperature efficiency coefficients,such as perovskite solar cells. Photons with lowerenergies are transmitted while photons with higher energies generatehot hole–electron pairs that relax by dissipating the heatenergy exceeding the energy gap. Further reduction of efficiency resultsfrom technological factors, including hole-pair recombination at defects.Larger conversion efficiencies may be obtained by multiple-junctionor tandem cells, pairing more absorbing materials. However, this unavoidablyleads to higher fabrication costs and complexity.Photovoltaic(PV) cells are devices capable of converting electromagneticradiation into electricity. The efficiency of a PV cell is intrinsicallylimited by the physical mechanism of power conversion11 although hybrid solar harvesters have not yet moved to manufacturing.Hybrid thermoelectric-photovoltaic(HTEPV) cells are a possibleway to enhance solar harvesting efficiency by  recoveringthe heat dissipated by the PV stage(s) through a thermoelectric (TE)stage. Over the last years, several approaches have been pursued toeffectively pair PV and TE stages. Laboratory prototypes of HTEPVgenerators have been fabricated and tested as well,The goal of this paper is to provide a comparative estimate ofpower costs for PV and hybrid solar harvesters built using contactTE-PV pairing with no reference to specific classes of TE and PV materials.An economic convenience index (EcCI) will be proposed to evaluateboth concentrated and nonconcentrated HTEPVs. Analysis of power costswill be complemented by an evaluation of the payback period (PBP)of hybrid solar harvesters. This work will focus on single-junctionsolar cells working at small solar concentrations, which are the dominatingtechnology in the domestic (rooftop) market. Since the analysis thatwill be proposed relies on the cost structure of the hybrid solarharvesters, multiple-junction solar cells will not be considered to keep the economic analysis manageable andcompendious. Additionally, with multiple-junction solar cells withPV efficiencies commonly exceeding 35%, making HTEPV harvesters profitablewould face the challenge of achieving efficiency improvements of severalpercentage points to pay back the additional costs related to theuse of segmented thermoelectric generators (TEGs) operating over extendedtemperature ranges.14 and opticalcoupling.15 However, while such pairingschemes avoid temperature compromises between PV and TE stages, theyinevitably make unavailable to the TE conversion the large amountof heat released by the PV stage. Furthermore, HTEPV harvesters arenot the only possibility to use sun power more extensively.Seemingly, direct (contact) pairing 1 might n16 van der Sark evaluatedthe affordability of pairing TEGs to PV cells by considering the extracost of a TEG retrofit sized in such a way to convert all heat releasedby the PV stage, reaching the conclusion that HTEPV could have attainedeconomic convenience only when TEG costs would have dropped down byat least a factor of 10. More recently, Zhu et al.17 computed the power costs χHTEPV (USD/W)of hybrid harvesters by optimizing costs ahead of power conversion.Economic convenience was properly assumed to occur when the powercost of the HTEPV was lower than that of the PV module, under theadditional requirement that the total power output also exceeded aminimum output power set by user-application needs. Unfortunately,Zhu et al. fully neglected the areal cost of installation. In addition,PV efficiency was taken as a constant, not depending on (decreasingwith) hybridization. This makes their results valid only for spectrum-split solutions.Beyond energetic convenience, for HTEPV harvesters to be a viabletechnology, their economic profitability also needs to be assessed.Possibly, the first attempt to estimate HTEPV convenience was advancedby van der Sark.18 Direct pairing was investigated, showing that, for current TE technologies(ZT ≈ 1), further to an improvement of efficiency, suitablechoice of heat exchangers makes HTEPV also economically viable. Theweight of heat exchanger costs was further considered by Rodrigo etal.,19 still investigating highly concentratedsolar harvesters and triple-junction solar cells. While high-efficiency,yet unavailable, TEGs disclose excellent opportunities of enhancedconversion, both energetically and economically, a trade-off was searchedbetween cost and efficiency by optimizing the concentration factor,the heat sink thermal resistance, and the TEG area. A predicted costreduction of about 38% is found at 1900 suns for realistic TEG figuresof merit. Unfavorable conclusions were reached instead for nonconcentratedSi-based PV cells20 where, compared tostand-alone PVs, an optimal levelized cost of energy higher by 8.7–90%was computed.In more recent years, several papershave been published reportingaccurate evaluations of the economic viability of TE hybridizationof given classes of PV modules. Integration of the concentrated PVsystem based on triple-junction cells (GaInP/GaInAs/Ge) with TEGswas analyzed by Rezania and Rosendahl.21 accounting forthe concurrent cost increase due to the TEG stage and the decay ofPV performance due to the heating of the PV stage based upon the physicalcharacteristics of PV and TE materials only. In this paper, we willfully develop such a methodology. Its main merit is that it makeseconomic analyses an additional tool to be used to scout suitablePV-TE material pairs for HTEPV generators. Although the design ofefficient HTEPV generators critically depends on a number of factors,22 active materials obviously play a pivotal role.We anticipate that the PV material governs the power effectivenessof any hybridization. As mentioned, in most hybridization schemes,the PV cell temperature raises, so the pairing of a PV stage witha TE stage implies either compromises or innovative layouts to befound. While the PV efficiency is commonly large at lower temperatures,TEGs require high temperatures at their hot side and large temperaturedifferences across their legs. This notwithstanding, inorganic PVmaterials may disclose realistic opportunities of successful hybridizationleading to increased total efficiencies. Specifically, performancesof large-gap PV materials are less sensitive to the increased temperaturesneeded by pairing with TEGs. Therefore, concentrated solar cells basedon a-Si, Cu2ZnSnS4 (CZTS), copper gallium selenideCuGaSe2, and GaInP may be sensibly expected to be suitablefor hybridization, along with stabilized perovskite solar cells. Instead,this rules out some common PV systems .Furthermore, low-cost, nontoxic materials with acceptable ZTs at moderate-to-lowtemperatures are needed for the TE legs.A different approach to profitability evaluationwas suggestedby the present authors,23 and very low emittances fromleg lateral surfaces also remain important in HTEPV generators. Onthe PV side, effective heat mirrors limiting the upward radiativedissipation of heat were shown to remarkably enhance the overall efficiencyof TEGs.24 Furthermore, heat dissipationat the TEG cold side, playing a key role in any TEG, is a formidablechallenge still nowadays, calling for innovative materials and advancedsurface finishing.26 However, differently from PVand TE materials, such materials issues do not set conflicting requirementswhen designing HTEPV cells.Materials issues arehowever not limited to active parts of thetwo solar stages. As an example, suppressing thermal cross-talk amongTE legs is of paramount relevance in solar thermoelectric generators(STEGs),2Economicconvenience of a renewable power technology is set, ingeneral terms, by the capital cost of the generator, namely, the costof the harvester (including the electronics needed to make the electricpower output useable) and of its installation. Therefore, power cost(USD/W), defined as the electric power generated divided by the capitalcost of the plant, is the basis of comparison among competing renewableenergy technologies. Economic convenience of a power technology integratingan existing renewable power source should be evaluated both in comparisonto existing renewable technologies and with respect to all power sources,including nonrenewable ones.In what follows, the analysis ofcapital costs of HTEPV generatorswill be split into installation costs and costs of the HTEPV components.Power cost is however only a part of the analysis. PBP measuresthe profitability of a renewable power technology compared to theoverall market of power sources (including nonrenewable). PBP is definedas the period of time needed for the energy obtained by the powerplant to compensate the plant capital costs. Therefore, PBP dependsupon the current energy price. For a technology to be profitable,PBP must be shorter than the average lifetime of the power plant.Therefore, PBP will also be evaluated.2.130 Among them, the approach developed by Yee et al.30 is especially suitable for the forthcoming analysis ofthe HTEPV economic convenience. As observed, optimization of a TEGon its efficiency followed by the search for the lowest possible systemcost forces the analysis to pursue the smallest possible cost at thehighest peak power while neglecting scenarios where lower peak powersmight lead to lower power costs. Therefore, the overnight capitalcosts were evaluated. Here and in the rest of this paper, we willuse double and triple primed symbols to refer to areal and volumecosts, respectively. Volumetric module costs CTEG‴ (USD/m3), i.e., costs of TEG components scaling with the module volume, were combined with areal module costs CTEG″ (USD/m2), i.e., costs of TEG components scaling withthe module contact area , and with heatexchanger costs cHX (USD/(W K–1)). Total TEG cost CTEG accounted toF isthe filling factor, defined as the ratio between the total TE legarea and the TEG footprint area STEG, L is the leg length, and U is the heattransfer coefficient. The power output PTEG reads30pn ≡ αp – αn (with αp andαn being the Seebeck coefficients of the p and nlegs), and ΔT is the temperature differencebetween the two heat reservoirs. Therefore, The analysisof the economic sustainability of TEGs has been the subject of severalpapers.EG″ USD/m, i.e., cF and UL/κ. Nonetheless, the χTEG surface displaysa narrow region around the line F = UL/(2κ) where χTEG takes low values, resultingfrom a competition between costs and TE performances.30 For smaller L (at constant F), costs decrease along with the temperature drop across the device,so that power output also decreases. Furthermore, a characteristicpoint existsL and F has only marginal benefits on χTEG. Exemplar values for CTEG″, CTEG‴, and CHX lead to optimal χTEG around60 USD/W, including the large contribution arising from the heat exchanger.Yee et al. showed that CHX = 0 in TEG maybe achieved for leg lengths L shorter than the thermal-impedance-matchingvalue 2Fκ/U, showing how morefavorable power costs may be obtained for TEGs operating under nonoptimizedconditions. Economic convenience stems from lower material costs overcompensatingthe reduced power output.Of special relevance to hybrid solar generators is the scenariowherein no heat exchanger cost adds up. Thus 2.22 for site preparation , mounting, and structural installation,with the complement to the BoS due to business costs.31 Business costs will be disregarded in what follows, sincewe focus on overnight capital costs only. Total costs and cost structuresquite differ for other PV materials, although module costs quite lineup to polycrystalline silicon modules. For CuInxGax1–Se2 (CIGS),module costs are reported to range from 0.63 to 0.69 USD/W, whileCdTe modules are quoted at 0.49 USD/W.32 Slightly lowercosts imply lower efficiencies  thatrequire larger module areas per output watt. In the forthcoming analysis,we will split BoS costs as costs per watt χBoS,1  andareal costs CBoS,2″ .Coststructure of PV plants, a current deployed technology, is paradoxicallymore complex to analyze than that of TEGs, namely a forthcoming technology.This is basically due to the rapid changes of the cost structure,redistributing costs across diverse technology elements. We will focusthis analysis on rooftop (domestic) solar plants, with typical poweroutputs below 20 kW. In this class of harvesters, PV capital costsmay be split into four main components, namely, PV materials, substrates(commonly glass), labor, and OEM costs. The dominating technologybased on polycrystalline silicon reports rooftop module costs of 0.85USD/W, to which about 1.00 USD/W must be added due to the balance-of-system(BoS) costs, accounting to 0.12 USD/W for the inverter, 0.08 USD/Wfor wiring and transformers, 0.05 USD/W for electrical installation,0.002 USD/m2.3Based on the frameworkset up for TEG and PV modules, economicconvenience of hybrid solar harvesters may be evaluated.33 This was clearly proved for STEGs, where improvements of power outputsby a factor of 3 were achieved by accurately minimizing thermal shuntsin the generator.34 Therefore, any economicestimation based upon computational models might easily turn out tobe overoptimistic, especially in view of the low technological maturityof HTEPV technologies. Furthermore, TEG market is still modest andmostly limited to Bi2Te3-based systems, so thatcost estimates might turn out to be prospectively inaccurate. Thissaid, hybrid harvesters must meet two requirements to be profitable.On the one side, their power costs must be lower than the power costof their associated PV technology, so that hybridization leads toan economic advantage for the final user. Furthermore, and independently,power costs must also be competitive with those of leading renewabletechnologies, currently set forth by polycrystalline silicon (pcSi)PV plants.35A fulleconomic analysis of HTEPV generators is a very complex task for severalreasons. First of all, layouts to pair PV and TE stages are extremelydiversified, encompassing direct thermal contact, thermal concentration,and solar-splitting strategies. In addition, power output from HTEPVcritically depends on many subtle constructive details.36 This is a key point that mustbe stressed. Often, radiative dissipation is disregarded in the thermalbudget analysis. In the absence of heat mirrors, instead, a majoramount of heat is irradiated, even at relatively low temperatures, that acts as a black body converting all electromagnetic radiationtransmitted by the PV cell. We further assume that the whole devicebe suitably encapsulated, so as to prevent convective heat dissipation.Furthermore, radiative dissipation from the PV cell is fully blockedby a suitable heat mirror, namely, a coating on the part of the encapsulatingpackage facing the sun that reflects electromagnetic radiation emittedby the PV cell (mostly in the infrared range) while transmitting solarradiation. and ref ). Theref1.CBOS, will be split in two parts: one, roughly proportionalto the power output, will account for the electronics , while costs encompassingsite preparation, mounting, and installation will be set proportionalto the module footprint. ThusThe costs of the BoS, G is the solar inputpower density, γ is the optical concentration, and ηHTEPV is the efficiency of the HTEPV harvester;2.SPVThe cost of the PVcell will scalewith the nonhybridized PV power output, that is, with its area PV is the PV costper watt and ηPVM is the efficiency of the PV module;3.The costof the TE stage will be computedin view of its areal and volume costs through 4.SSSAAdditional costs will arise from theSSA, scaling with its area 5.U and the technology it is based upon, namelyCosts for the heatexchanger will dependon the heat flux to be dissipated through its heat transfer coefficient SHX is itsarea and cHX units are USD/(W/K).To compute the projected power costs of HTEPV harvesters, we willmove from the following assumptions.SPV = STEG = SSSA = SHX ≡ S2Te3 TEGs. The effect of their variations for alternate materials willbe analyzed in Therefore, assuming hereafter 3In this section, efficiencyand power output for hybridized (HTEPV)and nonhybridized (PV) solar harvesters will be computed. We wouldlike to stress once again that the present effort to compute poweroutputs for a yet-to-be technology is unavoidably frustrated by manyfactors ruling the actual efficiency of hybrid solar harvesters. Thus,the computations we propose are to be meant as a best-case evaluationfor domestic (rooftop) solar harvesters. Therefore, they should beused as a no-go criterion, namely, showing when pairing between PVand TE stages may not be profitable, even in the absence of any factorfurther degrading the efficiency gain.3.1T with the heat flux throughthe TEG. Neglecting any lateral heat dissipation (due to the encapsulation)as well as the  fraction of heat converted into electric powerby the TEG,39 the temperature drop acrossthe TEG accounts to ϕ′/KTEG, where ϕ′ is the heat flux input from the PV stageinto the TEG and KTEG is the thermal conductanceof the TEG. It simply computes to22 . Therefore, ϕ′ is a best-case estimation of the realheat flux converted by the TEG, in accordance with the general purposeof this work.To estimate thepower output of the HTEPV harvester, it is convenient to rewrite theTEG power output N legs of length L and cross-section A, if κ is thethermal conductivity of the legs ,thenF = NA/S. The power outputof aTEG is PTEG = αpn2ΔT2/(4RTEG) where the electrical resistance RTEG reads 4L/(σSF), with the factor 4 accounting for the series electrical connectionof the legs.30 Thus30 since here the computationis carried out by setting the temperature of the hot side along withthe heat flux at the cold side. It is also remarkable that the poweroutput is formally independent of the thermal resistance of the heatexchanger, which enters instead to set the temperatures T1 (and T2) of the hot andcold sides of the TEG.For a TEG made of Eg, and its technologicalreadiness is computed according to standard models (cf. Supporting Information for details). It accountstoz ≡ eγEREΦ↑/r0, ERE is the external radiative efficiency,40e is the Neper’s number, r0 is the radiative recombination eq S1, Φ↑ is the fluxof photons with E > Eg at 1 sun eq S6, kB is the Boltzmann constant, and W(z) is the Lambert function. Note that ηPV depends on TPV. Replacing it into TPV – Ta (where Ta is ambient temperature) may be obtained byusing Fourier equation under the sensible assumption that the temperatureof the PV cell is equal to that of the hot side of the TE stage, namely, TPV = T1. SinceKtot–1 = KTEG–1 + KHX–1), replacing KHX with US, one obtainsTPV.The efficiency of the single-junctionPV stage along with its dependenceon γ, PHTEPV = PPV + PTEG = ηHTEPVSGγand leads toAssuming that each generator providesits output to an independent,optimized electric load, the total power output reads 3.2TPV is lower than in the hybriddevice due to the lack of the TEG and its contribution to the totalthermal resistance.Forthe standard PV module, computations simplify. Fourier equationreads in this case437 one of the presentauthors introduced an energetic convenience index to evaluate theenergetic advantage of pairing PV and TEG generators into a hybridsolar harvester. In what follows, an EcCI will be defined and analyzedto establish the economic profitability of such a coupling.In a previouspaper,F = 0.1, L = 0.1 cm, and ηHTEPV = ηPVM = 0.2, one may note how the dominating cost factors are in all casesthose of components which are not directly related to the harvesters,since BoS and the heat exchanger mostly set the power costs.Before proceeding to the analysis of the EcCI, some preliminarycomments may be in order. CBoS,2″.The power cost (USD/W) of the HTEPV harvester reads eq The corresponding power costfor the nonhybridized PV module isinstead eq An EcCIis defined as55.1Z, Eg, and the ERE),on constructive specifications , and on the operative conditions (γ).Optimization (maximization) of EcCI on the constructive parametersleads therefore to maximal profitability. It is easy to verify thatχHTEPV (L/(κF). In most cases, however, it is technology to set limitsto both L and F values since leglengths in excess of 1 cm are possibly too critical to manufactureand use, and filling factors larger than 0.1 would induce large radiativeinterleg cross-talking.30 Furthermore, EcCI is found to admit a maximumfor large U values. However, such values largelyexceed any realistic capability of heat dissipation by conventionalexchangers. Thus, a limit of 104 W/m2K has beenstipulated in all computations. As a result, optimized EcCI ultimatelydepends on the materials parameters and on solar concentration only.Consequently, we split optimal EcCI analysis into two major cases,namely, concentrated and nonconcentrated harvesters.TheEcCI is a multivariable function depending on material parameters(tχHTEPV 24 is a d5.2Z = 0.004 K–1). Beyond the real availability of TEGs with suchhigh performances, in all cases, the increase of economic profitabilityis truly marginal. For realistic Z values of ≈0.003K–1, EcCI is larger than unity only for Eg > 2.2 eV in technologically mature PV materials(ERE = 1 × 10–2). For less optimized PV absorbers,instead, EcCI exceeds 1 only for Eg >2.4 eV. This is sensible since for low EREs, the heating of the PVstage is more severe, and larger energy gaps are needed to make thereduction of PV efficiency less severe. The dominating role of thePV stage at low EREs and current Z values is confirmedconsidering that, instead, at  large Zs, EcCI >1 for almost any Eg value since extremely efficient TEGs would enable TE power generationovercompensating the decrease of PV efficiency.Inspection of For concentrated solar harvesters, analyses will be limited tothe case of γ = 5, a typical concentration rate for rooftopsolar plants.–2) requires Eg > 2.1 eV. As in the nonconcentrated case, less-performingPV absorbers (ERE = 1 × 10–6) need insteadlarger energy gaps (>2.3 eV) to attain EcCI > 1.In concentrated generators, EcCI shows slightlylarger margins4. For cuZ = 0.003 K–1. Rather, a wide-gap PV materialsuch as CdS (Eg = 2.4 eV) would benefitfrom PV-TE pairing, with EcCIs ranging from 1.003  to 1.01 , reaching 1.026 and 1.050 forhigh ERE and PVs at γ = 1 and 5, respectively. It should bestressed that an EcCI of 1.05, although seemingly meager, is insteadequivalent to an efficiency enhancement of 5% of solar conversionefficiency at a constant price. Thus, although profit margins arelimited, one may conclude that rooftop concentrated solar harvestersmight be anyway a possible driver to support the initial developmentof the HTEPV technology using novel, low-cost wide-gap PV materials—withan advantage when moderate solar concentration is considered. Thisis especially interesting if one considers that rooftop panels areamong the most rapidly expanding market niches in the sector of renewableenergies and that profitability may be eventually enhanced by triplecogeneration, with sanitary hot water being made available in additionto electric power production.41For polycrystalline silicon, a key player in the PV technology,it follows that in all cases, one computes EcCI < 1 for 5.3It is interestingto consider how a prospective cost rebate ofsingle parts of the technologies involved in HTEPV would impact theEcCI.CTEG‴) would lead to an absolute EcCIimprovement of ≈0.004. Even smaller is the impact of a similarreduction of areal costs (CTEG″), causing EcCI to increase by≈0.001. In both cases, changes of EcCI are comparable independentlyof the PV energy gap, ERE, and solar concentration. It is insteadpossibly unexpected that savings up to 50% of exchanger costs perW/K (cHX) would worsen EcCI—byabout 0.04 for γ = 1 and by about 0.005 for γ = 5. Thisis basically due to the fact that savings are more relevant to PVmodules. Thus, even if HTEPV costs would obviously decrease, nonethelessthey would decrease more in nonhybrid PV modules, and therefore EcCIwould worsen. Stated differently, a rebate of the exchanger cost wouldimprove EcCI only if the TE module had a significantly larger efficiency.Numerical estimates show that reductions up to 50% ofvolumetric module costs  display a remarkably differentdependence of their efficiency upon temperature and solar concentrationas the underlying physics of solar conversion in PSC somewhat differsfrom ordinary solar cells.46 have shown that thePSC efficiency, instead of decreasing, increases with temperature,reaching a maximum in the temperature range 45–55 °C,then rapidly dropping at higher temperatures. This is consistent witha tetragonal-to-cubic phase transition reported for methylammoniumlead iodide perovskites.49 Furthermore, the efficiency wasshown to increase with optical concentration (from 1 to 5 suns).50 Thus, EcCI was computed using data for a specificPSC50 adding the constraint that TPV < 323 K. Z values. Since estimates of cost parameters for PSCs arelargely variable, we inherited the same cost parameters used for thewhole analysis, namely, those typical of silicon solar cells  by anticipatingtheir commercial viability to a widely expanding market as that ofdomestic solar harvesting.The overall landscape we outlined should address researchalongtwo directions. Further to the prospective that would be opened bythe availability of TEG modules with higher efficiencies, HTEPV couldenable the concurrent use of TEs and low-efficiency, lower-cost PVmaterials, promoting at one time the diversification of PV materialsand reopening application interests toward age-old materials suchas On the other side, perovskites—andpossibly other new PVmaterials showing a temperature range wherein the PV efficiency remainsconstant or even increases with temperature—open great prospectsfor a beneficial and cost-effective TE hybridization.


We report molecular evidence of Tula virus infection in an immunocompetent patient from Germany who had typical signs of hantavirus disease. Accumulating evidence indicates that Tula virus infection, although often considered nonpathogenic, represents a threat to human health. Hantavirus disease, also called hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome, is a zoonosis; hantaviruses are transmitted from their reservoirs (rodents) to humans. The clinical course is characterized by initial high fever and body pain, potentially proceeding to renal, pulmonary failure, or both. The case-fatality rate depends on the causal virus species and can reach up to 50%  (Tula virus (TULV), a member of the family We report molecular evidence of TULV infection in a 21-year old immunocompetent man who originated from a small village near Hamburg, northern Germany. He was admitted to hospital for sudden fever, sickness, severe headache, abdominal pain, and limb pain since the day before. His medical history was unremarkable. The patient worked as a sanitary and heating engineer in the northern part of Germany. Except for elevated body temperature, the physical examination did not reveal any abnormalities. Blood testing at the day of admission revealed thrombocytopenia (63 platelets/nL), markedly elevated C-reactive protein (63.6 mg/L), and borderline leukocyte (9.8 cells/nL) and serum creatinine (1.2 mg/dL) values. Because of biochemical signs of infection, ultrasound findings of a moderate enlargement of spleen and cervical lymph nodes as well as hints of a small lung infiltration, an antibiotic regime (ampicillin/sulbactam and clarithromycin) was initiated. Over the next few days, the fever resolved.At day 4, biochemical signs of disturbed retention function indicated acute kidney injury. An enhanced serum creatinine value (1.6 mg/dL) and impaired (59 mL/min) glomerular filtration rate (GFR), determined using the CKD-EPI method, were noted; thrombocytopenia and an elevated C-reactive protein level persisted. The GFR, determined using the cystatin method, as measured at day 5, was reduced to 48 mL/min. The patient did not observe signs of oliguria. Under calculated intravenous fluid intake, the creatinine, GFR, and platelet values normalized until day 8, and the C-reactive protein declined to 12.1 mg/L. The patient was discharged from hospital at day 8 in good general condition.https://www.euroimmun.com). This assay does not contain TULV antigen, but the patient’s serum showed strong IgG reactivity with Dobrava–Belgrade virus (DOBV), Hantaan virus (HTNV), and Puumala virus (PUUV) antigens as well as strong band intensity on PUUV in the IgM assay. The recomLine HantaPlus IgG and IgM assays  revealed strong IgG reactivity to PUUV and in the IgM blot strong band intensities on PUUV, Sin Nombre virus, and DOBV. On the basis of these serologic findings, a PUUV infection of the patient was suspected.Initial laboratory diagnostics were based on hantavirus serologic test results  using the Hantavirus Profile 1 immunoblot  reacted exclusively with TULV diagnostic antigen (Myodes glareolus) and is a well-known pathogenic agent broadly distributed in Europe and parts of Asia  and mass reproduction of common voles in several parts of Europe, TULV should be considered as a threat to human health.The ability of TULV to cause hantavirus disease even in a previously healthy person shows the pathogenic potential of this virus. Therefore, the vole species hosting TULV should be considered as infection sources in their respective geographic ranges. Because of the broad distribution (