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Kaposi Sarcoma-associated Herpesvirus (KSHV) is the etiologic agent of Kaposi Sarcoma (KS), yet the viral genetic factors that lead to the development of KS in KSHV-infected individuals have not been fully elucidated. Nearly all previous analyses of KSHV genomic evolution and diversity have excluded the three major internal repeat regions: the two origins of lytic replication, IR1 and IR2, and the LANA repeat domain (LANAr). These regions encode protein domains that are essential to the KSHV infection cycle but have been rarely sequenced due to their extended repetitive nature and high GC content. The limited data available suggest that their sequences and repeat lengths are more heterogeneous across individuals than in the remainder of the KSHV genome. To assess their diversity, full-length IR1, IR2 and LANAr sequences, tagged with unique molecular identifiers (UMI), were obtained by Pacific Biosciences Single Molecule Real-Time sequencing (SMRT-UMI) from twenty-four tumors and six matching oral swabs from sixteen adults in Uganda with advanced KS. Intra-host single nucleotide variation involved an average of 0.16% of base positions in the repeat regions, compared to a nearly identical average of 0.17% of base positions in the remainder of the genome. Tandem repeat unit (TRU) counts varied by only one from the intra-host consensus in a majority of individuals. Including the TRU indels, the average intra-host pairwise identity was 98.3% for IR1, 99.6% for IR2 and 98.9% for LANAr. More individuals had mismatches and variable TRU counts in IR1 (12/16) than in IR2 (2/16). There were no open reading frames in the Kaposin coding sequence inside IR2 in at least fifty-five of ninety-six sequences. In summary, the KSHV major internal repeats, like the rest of the genome in individuals with KS, have low diversity. IR1 was the most variable among the repeats, and no intact Kaposin reading frames were present in IR2 of the majority of genomes sampled.

In this otherwise excellent article (Hoornweg et al., 2020) cited “Three human SEOV infections were confirmed, of which one was previously described as the first [2018] proven SEOV case in The Netherlands, based on IFT [immuno-fluorescence test] serology and an epidemiological link to SEOV RNA-positive [wild] rats) (Swanink et al., 2018), and now confirmed by comparative VNT [virus neutralization test].” This is historically incorrect: 27 years before Swanink et al, Groen et al. already reported wild rat-induced human SEOV infections in The Netherlands, moreover, in the same Dutch Institute of Public Health and Environmental Protection, Bilthoven, and with the same IFT (or IFA) technique (Groen et al., 1991). In addition, this 1991 report compared 14 non-laboratory cases of hemorrhagic fever with renal syndrome (HFRS), caused by the common European arvicolid orthohantavirus Puumala (PUUV), to 13 cases infected by the then rare (at least in early 1990s Western literature) orthohantavirus Seoul (SEOV), a juxtaposition in one single paper of two different local Dutch pathogenic orthohantaviruses, thus constituting at that moment the earliest such confrontation in nascent Western hantavirus literature (Figure 1). Moreover, not only 11 Dutch and/or Belgian laboratory rat-induced early 1980s SEOV-HFRS cases were detected, but also two wild rat-induced cases (arrows in Figure 1), thereby scoring yet another “first” in western hantavirus literature. Open in a separate window Figure 1 IFA: immuno-fluorescence assay or immuno-fluorescence test (IFT). ELISA, enzyme-linked immunosorbent assay. CTB ELISA, complex-trapping blocking, an inhibition ELISA variant. Closed circles: 1980s laboratory rat-acquired human hantavirus infections. Open circles: non-laboratory rat-acquired or “wild” human hantavirus infections, mostly being 1980s PUUV-induced, except for two “wild” cases (arrows), being 1980s wild rat-induced SEOV infections. Thick arrow: Dutch farmer, thin arrow: Belgian homeless vagabond, both with wild rat-exposure. In these two cases, SEOV infection was missed in IFA, showing surprisingly high cross-reacting PUUV titers (A), but confirmed, albeit with lower titers, in ELISA (B), and unmistakably ascertained in CTB-ELISA (C). Consequently, these two cases would have been mistaken for PUUV infections, if relying only on IFT/IFA. Adapted from Groen et al., 1991. Copyright © 1991 Wiley‐Liss, Inc., A Wiley Company.

Rat-borne Seoul hantaviruses are only just being recognized globally as important human pathogens, even though they have been studied and isolated on 4 different continents (Asia, Europe, the Americas, and Africa) since the early 1980s. Recent reports from Europe and the USA described Seoul hantavirus infection in pet rats and their breeders or owners, suggesting the potential emergence of a “new” public health problem. Wild and laboratory rat-induced Seoul hantavirus infections have, however, been described since the early eighties, due to the omnipresence of the rodent reservoir, the brown rat Rattus norvegicus. Recent studies showed no fundamental differences between pathogenicity and phylogeny of pet rat-Seoul hantaviruses and their formerly described wild or laboratory rat counterparts. The paucity of diagnosed Seoul virus-induced disease in the West is in striking contrast to the thousands of cases recorded since the 1980s in the Far-East, particularly in China. This review of four continents (Asia, Europe, America, and Africa) puts this “emerging infection” in historic perspective, concluding there is an urgent need for greater medical awareness of Seoul virus-induced human pathology in many parts of the world. Given the mostly milder and atypical clinical presentation, sometimes with preserved normal kidney function, the importance of simple urine examination is stressed, since initial, massive but transient proteinuria and microhematuria are rarely lacking. With increasingly sophisticated technology and enhanced clinical awareness, many specific hantaviruses are now recognized; however, SEOV remains the globally most widely distributed hantavirus.&nbsp

The 2019 11th International Conference on Hantaviruses (ICH 2019) was organized by the International Society for Hantaviruses (ISH), and held on September 1-4, 2019, at the Irish College, in Leuven, Belgium. These ICHs have been held every three years since 1989. ICH 2019 was attended by 158 participants from 33 countries. The current report summarizes research presented on all aspects of hantavirology: ecology; pathogenesis and immune responses; virus phylogeny, replication and morphogenesis; epidemiology; vaccines, therapeutics and prevention; and clinical aspects and diagnosis. ispartof: ANTIVIRAL RESEARCH vol:176 ispartof: location:Netherlands status: published

Although Seoul orthohantavirus is the only globally spread hantavirus pathogen, few confirmed human infections with this virus have been reported in Western countries, suggesting lower medical awareness of the milder, transient, and often chameleon-like symptoms of this zoonosis. We describe lesser known clinical and laboratory characteristics to help improve underreporting of this virus. ispartof: EMERGING INFECTIOUS DISEASES vol:25 issue:2 pages:387-388 ispartof: location:United States status: published

Climate change and land-use change are two major drivers of vegetation change causing habitat and biodiversity loss and posing a threat to the sustained provisioning of ecosystem goods and services. Following-up on the Millennium Ecosystem Assessment, the Sustainable Development Goals have been a fresh stimulus to the current interest in ecosystem services. Dynamic Global Vegetation Models (DGVMs) offer the possibility of integrating large amounts of geospatial data to quantify and project a large range of ecological variables important for ecosystem service provisioning under future scenarios. We outline how such model output could be used for projecting ecosystem service provisioning.

Intra-host tumor virus variants may influence the pathogenesis and treatment responses of some virally-associated cancers. However, the intra-host variability of Kaposi sarcoma-associated herpesvirus (KSHV), the etiologic agent of Kaposi sarcoma (KS), has to date been explored with sequencing technologies that possibly introduce more errors than that which occurs in the viral population, and these studies have only studied variable regions. Here, full-length KSHV genomes in tumors and/or oral swabs from 9 Ugandan adults with HIV-associated KS were characterized. Furthermore, we used deep, short-read sequencing using duplex unique molecular identifiers (dUMI)–random double-stranded oligonucleotides that barcode individual DNA molecules before library amplification. This allowed suppression of PCR and sequencing errors to ~10−9/base as well as afforded accurate determination of KSHV genome numbers sequenced in each sample. KSHV genomes were assembled de novo, and rearrangements observed were confirmed by PCR and Sanger sequencing. 131-kb KSHV genome sequences, excluding major repeat regions, were successfully obtained from 23 clinical specimens, averaging 2.3x104 reads/base. Strikingly, KSHV genomes were virtually identical within individuals at the point mutational level. The intra-host heterogeneity that was observed was confined to tumor-associated KSHV mutations and genome rearrangements, all impacting protein-coding sequences. Although it is unclear whether these changes were important to tumorigenesis or occurred as a result of genomic instability in tumors, similar changes were observed across individuals. These included inactivation of the K8.1 gene in tumors of 3 individuals and retention of a region around the first major internal repeat (IR1) in all instances of genomic deletions and rearrangements. Notably, the same breakpoint junctions were found in distinct tumors within single individuals, suggesting metastatic spread of rearranged KSHV genomes. These findings define KSHV intra-host heterogeneity in vivo with greater precision than has been possible in the past and suggest the possibility that aberrant KSHV genomes may contribute to aspects of KS tumorigenesis. Furthermore, study of KSHV with use of dUMI provides a proof of concept for utilizing this technique for detailed study of other virus populations in vivo., Author summary Kaposi sarcoma (KS) is a leading cancer in sub-Saharan Africa and in persons with HIV co-infection. Kaposi sarcoma-associated herpesvirus (KSHV, also referred to as human herpesvirus-8, or HHV-8) is the etiologic agent of KS, but the factors that contribute to the development of KS, which occurs in only a small subset of infected individuals, remain largely unknown. While strain differences or mutations in other tumor viruses are known to affect the risk and progression of their associated cancers, whether genetic variation in KSHV is important to the natural history of KS is unclear. Most studies of KSHV diversity have only characterized ~4% of its 165-kb genome, and the observed variation in some studies is likely to have been impacted by PCR or cloning artifacts. To precisely define genomic diversity of KSHV in vivo, we evaluated full-length viral genomes (except the internal repeat regions) using a technique that greatly lowers sequencing error rates and thus measures genomic diversity much more accurately than previous studies. In addition, we extended our analyses to look for potential tumor-specific changes in the KSHV genomes by examining viruses in both tumor and non-tumor tissues. To these ends, we performed highly sensitive, single-molecule sequencing of whole KSHV genomes in paired KS tumors and oral swabs from 9 individuals with KS. We found that KSHV genomes were virtually identical within the 9 individuals, with no evidence of quasispecies formation or multi-strain infection. However, KSHV genome aberrations and gene-inactivating mutations were found to be common in KS tumors, often impacting the same genes and genomic regions across individuals. Whether theses mutations influence KS tumorigenesis or result from genomic instability commonly found in tumors warrants further study. Lastly, aberrant KSHV genomes were found to be shared by distinct tumors within individuals, suggesting the capacity of KS tumor cells to metastasize and seed new lesions.

Climate change and land-use change are two major drivers of biome shifts causing habitat and biodiversity loss. What is missing is a continental-scale future projection of the estimated relative impacts of both drivers on biome shifts over the course of this century. Here, we provide such a projection for the biodiverse region of Latin America under four socio-economic development scenarios. We find that across all scenarios 5-6% of the total area will undergo biome shifts that can be attributed to climate change until 2099. The relative impact of climate change on biome shifts may overtake land-use change even under an optimistic climate scenario, if land-use expansion is halted by the mid-century. We suggest that constraining land-use change and preserving the remaining natural vegetation early during this century creates opportunities to mitigate climate-change impacts during the second half of this century. Our results may guide the evaluation of socio-economic scenarios in terms of their potential for biome conservation under global change.

Hantavirus infections, recently renamed 'hantavirus fever' (HTVF), belong to the most common but also most underestimated zoonoses in the world. A small number of reports described the so-called 'lipid paradox' in HTVF, i.e. the striking contrast between a very low serum total cholesterol and/or high-density lipoprotein cholesterol (HDLc), and a paradoxical concomitant hypertriglyceridaemia. In a prospective study, with patients being their own control after illness, we wanted to verify if this quick and easy 'bedside test' was robust enough to warrant a preliminary diagnosis of acute kidney injury (AKI) caused by HTVF. The study cohort consisted of 58 Belgian cases (mean age 44 years), admitted with varying degrees of AKI and of thrombocytopaenia, both characteristic for presumptive HTVF. All cases were sero-confirmed as having acute HTVF. At or shortly after hospital admission, a significant (p

Transient proteinuria and acute kidney injury (AKI) are characteristics of Puumala virus (PUUV) infection. Albuminuria peaks around the fifth day and associates with AKI severity. To evaluate albuminuria disappearance rate, we quantified albumin excretion at different time points after the fever onset. The study included 141 consecutive patients hospitalized due to acute PUUV infection in Tampere University Hospital, Finland. Timed overnight albumin excretion (cU-Alb) was measured during the acute phase in 133 patients, once or twice during the convalescent phase within three months in 94 patients, and at six months in 36 patients. During hospitalization, 30% of the patients had moderately increased albuminuria (cU-Alb 20&ndash, 200 &mu, g/min), while 57% presented with severely increased albuminuria (cU-Alb &gt, g/min). Median cU-Alb was 311 &mu, g/min (range 2.2&ndash, 6460) &le, 7 days after fever onset, 235 &mu, g/min (range 6.8&ndash, 5479) at 8&ndash, 13 days and 2.8 &mu, g/min (range 0.5&ndash, 18.2) at 14&ndash, 20 days. After that, only one of the measurements showed albuminuria (35.4 &mu, g/min at day 44). At six months, the median cU-Alb was 2.0 &mu, g/min (range 0.6&ndash, 14.5). Albuminuria makes a flash-like appearance in PUUV infection and returns rapidly to normal levels within 2&ndash, 3 weeks after fever onset. In the case of AKI, this is a unique phenomenon.

Hantavirus disease is a viral zoonosis, caused by inhalation of infectious aerosolized excreta from chronically infected rodents, which are both the reservoir and the vector of different hantavirus species. Hantavirus infections manifest mainly as haemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome, which traditionally but incorrectly were thought to be caused by exclusively Old World hantaviruses and New World hantaviruses, respectively.Hantavirus diseases are characterized by non-specific flu-like symptoms, followed by a sometimes lethal capillary leak syndrome, haemorrhage, and rarely by shock. Infection is accompanied by augmented release of pro-inflammatory cytokines which indirectly causes organ damage. Diagnosis can be made by serology or plaque reduction neutralization tests, detection of viral proteins by Western blot assay, or detection of hantavirus genome by reverse transcription-polymerase chain reaction. Treatment is mainly supportive.Together with leptospirosis, haemorrhagic fever with renal syndrome is the only form of acute kidney injury against which vaccines are in use, but a World Health Organization-licensed vaccine is still lacking.

Hantavirus disease or at least its renal form, the so-called haemorrhagic fever with renal syndrome is the only globally emerging acute kidney injury (AKI) form, and currently without doubt the most underestimated form of community-acquired AKI. Hantavirus disease is a viral zoonosis, caused by inhalation of infectious aerosolized excreta from chronically infected rodents, which are both the reservoir and the vector of different hantavirus species. Clinical presentation consists of sudden flu-like symptoms (fever, headache, myalgia), followed by gastrointestinal discomfort and AKI, often with anuria or oliguria. More rarely, acute myopia and/or non-cardiogenic acute lung oedema or injury is the presenting or complicating symptom. Laboratory hallmarks are initial thrombocytopenia and proteinuria, raised C-reactive protein and lactate dehydrogenase, left-shift leucocytosis, and typical but transient serum lipid disturbances. Spontaneous remission occurs within 2–3 weeks without sequelae. Case fatality rate is between 0.1% and 15% according to the infecting hantavirus species, but most infections show in fact an asymptomatic or paucisymptomatic presentation. Treatment is only supportive, but may necessitate life-saving intensive care techniques. Together with leptospirosis, haemorrhagic fever with renal syndrome is the only form of AKI against which different vaccines are in use, but a World Health Organization-licensed hantavirus vaccine is still lacking.

Genetic diversity, attributable to the low fidelity of reverse transcription, recombination and mutation, is an important feature of infectious retroviruses. Under selective pressure, such as that imposed by superinfection interference, gammaretroviruses commonly adapt their envelope proteins to use alternative receptors to overcome this entry block. The first characterized koala retroviruses KoRV subgroup A (KoRV-A) were remarkable in their absence of envelope genetic variability. Once it was determined that KoRV-A was present in all koalas in US zoos, regardless of their disease status, we sought to isolate a KoRV variant whose presence correlated with neoplastic malignancies. More than a decade after the identification of KoRV-A, we isolated a second subgroup of KoRV, KoRV-B from koalas with lymphomas. The envelope proteins of KoRV-A and KoRV-B are sufficiently divergent to confer the ability to bind and employ distinct receptors for infection. We have now obtained a number of additional KoRV envelope variants. In the present studies we report these variants, and show that they differ from KoRV-A and KoRV-B envelopes in their host range and superinfection interference properties. Thus, there appears to be considerable variation among KoRVs envelope genes suggesting genetic diversity is a factor following the KoRV-A infection process.

Background: Puumala hantavirus (PUUV) induces an acute tubulointerstitial nephritis and acute kidney injury (AKI). Our aim was to evaluate the prognosis of severe AKI associated with PUUV infection. Methods: We examined 556 patients who were treated at Tampere University Hospital during 1982-2013 for acute, serologically confirmed PUUV infection. Plasma creatinine was measured during hospitalization, convalescence, and 1, 2, and 5 years after the acute infection. Results: Plasma creatinine concentration was elevated (>100 μmol/l) in 459 (83%) patients, while altogether 189 patients (34%) had severe AKI defined as Kidney Disease: Improving Global Outcomes (KDIGO) stage 3, that is, plasma creatinine ≥353.6 μmol/l (4.0 mg/dl) or need of dialysis. There were no fatal cases during the hospitalization or the following 3 months. Fatality rate during the years following PUUV infection did not differ between patients who had suffered from severe AKI versus those without severe AKI. Post-hospitalization plasma creatinine values were available for 188 (34%) patients. One month after the acute infection, patients with prior severe AKI had higher median plasma creatinine concentration (82 µmol/l, range 54-184) than patients without severe AKI (74 µmol/l, range 55-109, p = 0.005). After 1 year, no significant difference existed in median plasma creatinine concentrations between patients with (71 µmol/l, range 36-123) and without prior severe AKI (72 µmol/l, range 34-116, p = 0.711). After 5 years all but 1 patient had normal creatinine levels. Conclusions: In contrast to the worldwide well-accepted KDIGO criteria, severe AKI associated with PUUV infection is not associated with excess fatality but has a very good prognosis, both in the short and long terms.

The European prototype of hantavirus, Puumala virus (PUUV), isolated from a common wild rodent, the bank vole (Myodes glareolus), causes nephropathia epidemica (NE). NE can perfectly mimic haemolytic-uraemic syndrome (HUS), progressing from an aspecific flu-like syndrome to acute kidney injury with thrombocytopaenia, and presenting with some signs of haemolytic anaemia and/or coagulopathy. Moreover, both NE and HUS can occur in local outbreaks. We report an isolated case of NE, initially referred for plasmapheresis for suspected HUS, although signs of overt haemolysis were lacking. Early suspicion of hantavirus infection, later confirmed by serology and reverse transcription polymerase chain reaction (RT-PCR), prevented subsequent excessive treatment modalities. ispartof: European Journal of Clinical Microbiology & Infectious Diseases vol:37 issue:1 pages:135-140 ispartof: location:Germany status: published

We examined the cause of death during a 12-month period (1995/96) in all consecutive patients admitted to hospital with leptospiral infection in Seychelles (Indian Ocean), where the disease is endemic. Leptospirosis was diagnosed by use of the microscopic agglutination test and a specific polymerase chain reaction assay on serum samples. Seventy-five cases were diagnosed and 6 patients died, a case fatality of 8%. All 6 patients died within 9 days of onset of symptoms and within 2 days of admission for 5 of them (5 days for the 6th). On autopsy, diffuse bilateral pulmonary haemorrhage (PH) was found in all fatalities. Renal, cardiac, digestive and cerebral haemorrhages were also found in 5, 3, 3 and 1 case(s), respectively. Incidentally, haemoptysis and lung infiltrate on chest radiographs, which suggest PH, were found in 8 of the 69 non-fatal cases. Dengue and hantavirus infections were ruled out. In conclusion, PH appeared to be a main cause of death in leptospirosis in this population, although haemorrhage in other organs may also have contributed to fatal outcomes. This cause of death contrasts with the findings generally reported in endemic settings.

Dobrava-Belgrade virus (DOBV) is a human pathogen that has evolved in, and is hosted by, mice of several species of the genus Apodemus. We propose a subdivision of the species Dobrava-Belgrade virus into four related genotypes – Dobrava, Kurkino, Saaremaa, and Sochi – that show characteristic differences in their phylogeny, specific host reservoirs, geographical distribution, and pathogenicity for humans.

ispartof: Eurosurveillance vol:21 issue:39 pages:27-28 ispartof: location:Sweden status: published

To the editor: The recent confirmation by RT-PCR of a case of haemorrhagic fever with renal syndrome (HFRS), induced by Tula virus (TULV) in France [1], confirms the pathogenicity of this arvicolid hantavirus, a fact not generally acknowledged yet, or at least still contested [2]. The clinical presentation of the demonstrated TULV HFRS case was, however, unusual: besides the classic fever with thrombocytopenia and elevated transaminases, leukopenia instead of leukocytosis with left shift was found, and the renal function remained strictly within normal limits. However, renal involvement was nevertheless indicated by transient microscopic haematuria. Regrettably, transient but massive and unselective proteinuria, the renal hallmark in probably all hantavirus infections, was once more not discussed. Interestingly, a false-positive serological screening result for another arvicolid hantavirus, Puumala virus (PUUV), was obtained in three assays of two different formats (immunofluorescence assay (IFA) and ELISA), but could not be confirmed by routine RT-PCR, i.e. by using PUUV-specific primers [1]. As subsequently shown by Reynes et al., TULV and PUUV are two closely related, yet genetically distinct hantavirus species, both carried by distinct voles of the Arvicolinae, a subfamily of the Cricetidae rodent family [1].

Impacts • Dynamic data-based model generates stochastic forecasts of the occurrence of nephropathia epidemica (NE) epidemics 3 months ahead. • We have demonstrated that NE outbreaks can be accurately predicted using climatic and vegetation data or bank voles' population dynamics with a dynamic data-based model. • Such a modelling approach can be used to develop new tools to help prevent future NE outbreaks. Summary Wildlife-originated zoonotic diseases in general are a major contributor to emerging infectious diseases. Hantaviruses more specifically cause thousands of human disease cases annually worldwide, while understanding and predicting human hantavirus epidemics pose numerous unsolved challenges. Nephropathia epidemica (NE) is a human infection caused by Puumala virus, which is natu- rally carried and shed by bank voles (Myodes glareolus). The objective of this study was to develop a method that allows model-based predicting 3 months ahead of the occurrence of NE epidemics. Two data sets were utilized to develop and test the models. These data sets were concerned with NE cases in Finland and Belgium. In this study, we selected the most relevant inputs from all the available data for use in a dynamic linear regression (DLR) model. The number of NE cases in Finland were modelled using data from 1996 to 2008. The NE cases were predicted based on the time series data of average monthly air tem- perature (� C) and bank voles' trapping index using a DLR model. The bank voles' trapping index data were interpolated using a related dynamic harmonic regression model (DHR). Here, the DLR and DHR models used time-varying parameters. Both the DHR and DLR models were based on a unified state-space estimation framework. For the Belgium case, no time series of the bank voles' population dynamics were available. Several studies, however, have suggested that the population of bank voles is related to the variation in seed production of beech and oak trees in Northern Europe. Therefore, the NE occurrence pat- tern in Belgium was predicted based on a DLR model by using remotely sensed phenology parameters of broad-leaved forests, together with the oak and beech seed categories and average monthly air temperature (� C) using data from 2001 to 2009. Our results suggest that even without any knowledge about hantavirus dynamics in the host population, the time variation in NE outbreaks in Finland

In the present issue of Infectious Diseases, an article by Zheng et al. considers the persistence of IgG antibodies (Abs) after hantavirus vaccination, and compares it to the persistence of IgG Abs...

The connection between nephropathia epidemica (NE) and vegetation dynamics has been emphasized in recent studies. Changing climate has been suggested as a triggering factor of recently observed epidemiologic peaks in reported NE cases. We have investigated whether there is a connection between the NE occurrence pattern in Belgium and specific trends in remotely sensed phenology parameters of broad-leaved forests. The analysis of time series of the MODIS Enhanced Vegetation Index revealed that changes in forest phenology, considered in literature as an effect of climate change, may affect the mechanics of NE transmission. ispartof: International Journal of Environmental Research and Public Health vol:7 issue:6 pages:2486-2500 ispartof: location:Switzerland status: published

Hantaviruses, members of the family Bunyaviridae, are the causative agents of hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome. Hantaviruses are currently demarcated into species based on the guidelines provided by the International Committee on Taxonomy of Viruses (ICTV). These guidelines however, are often ignored by the descriptors of novel hantaviruses. With this study we attempted to refine the second ICTV guideline for hantavirus species demarcation by phylogenetically analyzing all in Genbank available complete sequences derived from the S, M or L segments of hantaviruses. S and M segment amino acid sequence comparison allowed clear and unequivocal distinction between different hantavirus species, and lead us to propose additional criteria for the demarcation of hantavirus species (S segment amino acid distance >10% or M segment amino acid distance >12%) and hantavirus groups (S segment amino distance >24% or M segment amino acid distance >32%). With this study, we propose to adjust the second rule of the ICTV classification guidelines ("a 7% difference in amino acid identity when comparing the complete S segment and M segment sequences") to a more appropriate rule, "a 10% difference in S segment similarity and a 12% difference in M segment similarity based on complete amino acid sequences" in accordance with the current situation in the hantavirus field.

Hantaviruses, which are mainly rodent-borne viruses, cause hemorrhagic fever with renal syndrome in the Old World, and hantavirus pulmonary syndrome in the New World. A neutralization test based on quantitative RT-PCR, the replication reduction neutralization test (RRNT), was developed for efficient detection of hantavirus-neutralizing antibodies. The effectiveness of the RRNT was evaluated by examining several hantaviruses and hantavirus-specific convalescent human serum samples. All convalescent serum samples tested by RRNT caused significant decreases in hantavirus genomes with only one specific hantavirus species, which allowed a straightforward identification of the related hantavirus. The results obtained by RRNT were completely comparable with the results obtained by focus reduction neutralization test (FRNT). The RRNT approach is a reliable and rapid alternative for FRNT, hitherto considered as the gold standard for hantavirus serology.

(2007). ACUTE KIDNEY INJURY IN EMERGING, NON-TROPICAL INFECTIONS. Acta Clinica Belgica: Vol. 62, No. 6, pp. 387-395.

SUMMARYTwo global (re-)emerging zoonoses, leptospirosis and hantavirus infections, are clinically indistinguishable. Thirty-one patients, hospitalized in Sri Lanka for acute severe leptospirosis, were after exclusion of other potentially involved pathogens, prospectively screened with IgM ELISA for both pathogens. Of these, nine (29·0%) were positive for leptospirosis only, one (3·2%) for hantavirus only, seven (22·5%) for both pathogens concomitantly, whereas 13 (41·9%) remained negative for both. Moreover, in a retrospective study of 23 former patients, serologically confirmed for past leptospirosis, six (26·0%) were also positive in two different IgG ELISA hantavirus formats. Surprisingly, European Puumala hantavirus (PUUV) results were constantly higher, although statistically not significantly different, than Asian Hantaan virus (HTNV), suggesting an unexplained cross-reaction, since PUUV is considered absent throughout Asia. Moreover, RT–PCR on all hantavirus IgM ELISA positives was negative. Concomitant leptospirosis-hantavirus infections are probably heavily underestimated worldwide, compromising epidemiological data, therapeutical decisions, and clinical outcome.

A truncated recombinant nucleocapsid protein (rNp118) consisting of the first 118 amino-terminal amino acids (AA) of the Puumala hantavirus (PUUV) nucleocapsid protein expressed in Escherichia coli, was evaluated for its antigenicity and reliability as serodiagnostic antigen in an enzyme-linked immunosorbent assay (ELISA) for detection of PUUV antibodies in human sera. The PUUV nucleocapsid protein has been shown to contain several B-cell epitopes, mapped within the first 118 amino-terminal AA. This finding makes the rNp118 an interesting recombinant protein to use as serodiagnostic antigen. The sensitivity of this new PUUV-rNp118 ELISA, was compared with those of a commercially available PUUV ELISA assay and an home-made ELISA based on a recombinant whole nucleocapsid protein of PUUV. Eighty-six human serum samples clinically suspected for PUUV-induced nephropathia epidemica, and previously screened with the reference assays, were tested. The sensitivity of the new assay was compared with that of the reference assays and an excellent correlation between the assays was found. Sera found to be negative by other methods were also negative in our assay. The ELISA based on rNp118 represents an alternative and valid test for detection of antibodies to PUUV in human sera.

J Clement (jan.clement@uzleuven.be)1, M Van Esbroeck2, K Lagrou3, J Verschueren2, N P Sunil-Chandra4, M Van Ranst1 1. National Reference Centre for Hantaviruses, Clinical and Epidemiological Virology, University Hospitals of Leuven, Leuven, Belgium 2. National Reference Laboratory for Leptospirosis, Department of Clinical Sciences, Institute of Tropical Medicine, Antwerp, Belgium 3. Department of Microbiology & Immunology, Catholic University Leuven, and Clinical Department of Laboratory Medicine, University Hospitals of Leuven, Leuven, Belgium 4. Department of Medical Microbiology, Faculty of Medicine, University of Kelaniya, Sri-Lanka

Nephropathia epidemica (NE) is a human infection caused by Puumala virus (PUUV), which is naturally carried and shed by bank voles (Myodes glareolus). The objective was to develop a dynamic model of the NE cases and the bank vole population in both Finland and Belgium by defining the periodic components with a dynamic harmonic regression (DHR) model. The defined periodic components can be further used to adapt mechanistic Susceptible and Infective (SI) models regionally. Despite the difference in bank vole population dynamics and NE cases between the Western European temperate zone and boreal zones the DHR model was able to quantify the dynamics of NE cases in Belgium and Central Finland with a coefficient of determination (R2) of 0.70 and 0.82 respectively and to quantify the dynamics of bank vole population in Belgium and Central Finland with R2 of 0.80 and 0.98 respectively. DHR identified 18 month cycles in the bank vole population in Belgium. This approach demonstrated two year cycles in Belgian NE outbreaks. DHR identified three year cycles in Finnish bank vole populations which in turn cause three year cycles in the NE outbreaks in Central Finland. Because the bank vole population data in Finland was contemporary with the data of NE cases, the DHR showed a three month delay between the NE cases and the bank vole population in Central Finland. This approach may help us in our understanding of the spatial and temporal dynamics of NE cases and the bank vole populations in different regions.