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Widespread circulation of Alongshan virus in Austria and serological evidence for infection in humans: a nationwide molecular and serological observational study.

BACKGROUND: Alongshan virus, a segmented RNA virus in the Jingmenvirus group of flaviviruses, was first identified in 2017 in China in patients with tick-borne encephalitis-like illness. Alongshan virus has since been detected in ticks and mammals in several European countries. In this study, we aimed to characterise the temporal and geographical distribution of Alongshan virus in Austria through nationwide tick surveillance combined with serological and molecular screening in individuals with suspected tick-borne encephalitis or tick exposure. METHODS: In this nationwide molecular and serological observational study, we conducted a PCR-based screening of ticks collected across Austria in 2024, using flagging, animal hosts, and citizen submissions. We also collated genomic data from stored nucleic acid extracts from ticks collected in Austria in 2005 and 2013 in previous surveillance studies and stored extracts of paired tick-human samples collected between 2015 and 2018. Alongshan virus-positive tick samples were subjected to whole-genome sequencing and phylogenetic analysis. In addition to tick samples, blood samples from Austrian patients with reported tick bite or suspected tick-borne encephalitis, submitted to the National Reference Center for Human Arbovirus Infections, Austria, were screened for Alongshan virus infection and subjected to serological and molecular testing. FINDINGS: 2952 ticks were collected between Feb 1, 2024, and Dec 6, 2024, from 29 (83%) of 35 NUTS-3 regions in Austria; the median detection rate for Alongshan virus was 1·2% (IQR 0·4-3·5). In addition, 1816 archived tick samples were analysed, with three testing positive for Alongshan virus. For the virus-positive samples, phylogenetic analysis showed that sequences from Austria grouped within the European clade, with Austrian sequences from the same region showing high sequence similarity. 1361 human serum samples collected between March 1, 2023, and May 16, 2025, were assessed for anti-Alongshan virus IgG antibodies. Two individuals had high antibody titres against Alongshan virus proteins VP1a and VP2. INTERPRETATION: Our study shows detection of Alongshan virus in archived tick samples dating back to 2005, representing the earliest documented occurrence of the virus to date and suggesting that Alongshan virus has been circulating in Austria for at least two decades. The detection of Alongshan virus-specific antibody titres in two individuals suggests past infection and previously unrecognised exposure. These results highlight the need for continued molecular surveillance of tick populations and serological monitoring in humans to define the epidemiology and public health relevance of Alongshan virus in Europe. FUNDING: One Health surveillance and Vector monitoring for cross-border pathogens (OH SURVector) and UNITED4Surveillance.

Adolescent↗

Virus safety of human immunoglobulins: efficient inactivation of hepatitis C and other human pathogenic viruses by the manufacturing procedure.

Human immunoglobulins are plasma derivatives with a low risk of transmitting viral infections. To the present, no proven case of human immunoglobulins transmitting human immunodeficiency viruses has been reported. However, there have been a few reports on the transmission of hepatitis C virus by these plasma proteins. To improve further the safety of both 5s iv human immunoglobulins and 7s im immunoglobulins, we introduced a 10-hour heat treatment of the aqueous solutions at 60 degrees C (i.e., pasteurization) into the manufacturing procedure. This treatment was not added to the manufacturing procedure of 7s iv immunoglobulin that already contained the S-sulfonation as a virus inactivating method. We now report on experimental data that show that the whole manufacturing procedures of the above immunoglobulins inactivate efficiently hepatitis C virus and that the specific virus inactivation methods alone, namely, pasteurization or S-sulfonation, also inactivate completely viruses of the flavivirus family, to which the hepatitis C virus belongs. The inactivation of the Flaviviridae bovine viral diarrhea virus, tick-borne encephalitis virus, and yellow fever virus by pasteurization or S-sulfonation was at least 10(5). The clearance of HCV achieved by the entire manufacturing process of each of these immunoglobulins was also at least 10(5). The experiments therefore show that pasteurization or S-sulfonation provides a high margin of safety to human immunoglobulins regarding the transmission of hepatitis C virus.

Animals↗

Surveillance of tick-borne viruses in the border regions of the Tumen River Basin: Co-circulation in ticks and livestock.

BACKGROUND: The unique eco-geographical patterns and climatic conditions of the China-Tumen River border region, combined with frequent cross-border tourism and trade activities, collectively establish this area as a recognized hotspot for tick-borne disease outbreaks. However, critical knowledge gaps persist regarding the eco-epidemiology of emerging tick-borne viruses and the distribution of their potential reservoir hosts within this trinational ecosystem spanning China, North Korea, and Russia. METHODS: We collected a total of 2,004 ticks from the study area, along with blood samples obtained from 42 sheep and 45 cattle. Following viral metagenomic analysis of the ticks, dual verification of target pathogens in all samples was performed using qRT-PCR and RT-PCR assays. Phylogenetic trees were constructed and nucleotide sequences were analyzed to delineate relationships between the obtained virus strains and reference sequences. RESULTS: Viral metagenomics identified three viruses in ticks: Dabieshan tick virus (DBTV), Songling virus (SGLV), and Yanggou tick virus (YGTV). PCR analysis detected DBTV exclusively in Hunchun ticks (minimum infection rates, MIR:4.73%) and YGTV in Antu specimens (MIR:0.97%). Conversely, SGLV was detected in ticks from all four regions, with MIR of 1.68% (Helong), 0.74% (Hunchun), 1.61% (Antu), and 4.79% (Longjing). Concurrently, SGLV was detected in 19 sheep blood samples from Longjing, yielding a positivity rate of 45.24%, while YGTV was identified in 13 cattle blood samples from Antu, with a positivity rate of 28.89%. Phylogenetically, the DBTV strain clustered with previously reported DBTV and Yongjia tick virus 1 isolates. Sheep-derived SGLV strains shared close evolutionary ties with tick-borne SGLV, whereas YGTV from cattle and ticks formed a distinct cluster with Russian strains but diverged into two branches from Chinese variants, suggesting evolutionary instability. CONCLUSION: These findings address critical knowledge gaps in the transmission dynamics and genetic diversity of emerging arboviruses while providing vital insights for developing cross-border surveillance strategies with significant public health implications.

Animals↗

[Biological properties of tick-borne encephalitis strains isolated in different parts of its geographic range].

A comparative study of over 200 tick-borne encephalitis (TBE) virus strains isolated from patients and ticks in different regions varying in the proportion of severe clinical forms of TBE was carried out. The strains were tested for their pathogenicity for white mice, cytopathic and plaque-forming properties as well as for the interferon-inducing capacity in SPEV cells. All the strains were found to be highly neurovirulent for mice, to produce plaques with diameters varying form less than 1 to 10 mm, and to induce interferon production in SPEV cell. Among the TBE virus strains isolated in the Baltic republics and Vologda region there occurred strains causing complete destruction of SPEV cells at later intervals than the prototype strains. No association of the examined properties of TBE virus strains with the region of isolation or with the clinical forms of human disease was established.

Animals↗

The influence of specific antisera and unheated guinea pig serum on the pathogenicity of "Runde" virus for mice.

Antisera from various animal species containing antibodies to "Runde" virus were not able to neutralize virus infection in newborn mice, the outcome of which is an acute, fatal CNS disease. There was, however, one noticeable exception. Mixtures of virus and hyperimmune mouse serum or ascitic fluid inoculated intracerebrally into newborn mice resulted in a persistent infection and a chronic disease which had previously only been recognized in 2 to 3-week-old mice inoculated with "Runde" virus. A serum pool from persistently infected mice had the same effect, though this was less pronounced. The addition of unheated guinea pig serum to the virus-hyperimmune serum mixtures reinforced the tendency to persistence and chronic disease, and unheated guinea pig serum alone modified the infection in the same way. The results suggest an immunological basis for the virus persistence and chronic disease in suckling mice.

Age Factors↗

Experimental encephalitis in monkeys caused by the Powassan virus.

We have carried out a comparative study of the experimental infection of monkeys with the P-40 strain of the Powassen virus, isolated in the Primor'e Territory of the USSR, and with the Canadian prototype LB strain. The Powassan virus was found to be pathogenic for Macaca rhesus. The clinical and pathomorphological picture of the experimental encephalitis was studied, and the full identity of the infection produced in the monkeys by the P-40 strain and the Canadian LB strain of the Powassan virus was demonstrated. On electron microscopic examination of the central nervous system the virus was detected in the neurons, glial cells, and intercellular spaces. The virions of the strains studied have identical morphological parameters, being 37-45 nm in diameter and of spherical shape. The data obtained indicated a marked neurotropism of the virus. They will contribute to the elucidation of the role of the virus in the infection pathology of humans, i.e., in the differentiation of encephalitis cases not associated etiologically with the virus of the spring-summer tickborne encephalitis.

Animals↗

[Pathogenic factors in viral diseases of the central nervous system].

The possible immunological significance of the different kinds of cells participating in the inflammatory reaction in cerebrospinal infections is to be considered. Presence of plasma cells and occasional perivenous microglial seams in early stages of the neurotropic virus diseases must be regarded as immunological manifestations. The brainstem nuclei are sites of predilection both in entero- and in arbovirus diseases. The aetiologically different neurotropic virus diseases obtain their individual character from the involvement of certain neurones. Neural pathways are to be taken into consideration mainly for the propagation of enteroviruses, but may have a part also in the propagation of arboviruses. With massive invasion by highly cytopathogenic viruses inflammatory reaction may fail to appear in the immature nervous tissue. Some encephalitides, of both viral and non-viral aetiology, in which the prevalence of plasma cells in the infiltrations bears evidence of an intense immune process, may be interpreted as immunologically determined late diseases.

Animals↗

Adjuvant effect of human growth hormone with an inactivated flavivirus vaccine.

Vaccines made by inactivating pathogenic microorganisms have been dramatically successful in controlling diseases in humans and animals. Despite their successes, they have a major disadvantage in that several inoculations are required for them to be effective. To overcome this problem, a commercial inactivated vaccine preparation against tickborne encephalitis was combined with human growth hormone (HGH). This formulation produced complete protection in a murine model with only one dose of vaccine, apparently by binding hormone and antigen to an insoluble matrix containing aluminium hydroxide. Thus it is postulated that when virus-specific lymphocytes are attracted to the site of injection, the hormone is at a high local concentration and stimulates the clonal expansion of antigen-specific T cells. The development of genetically engineered HGH now gives unlimited supplies of hormone, potentially resulting in an increase in efficacy of a wide variety of vaccines, especially those needing prolonged immunization schedules such as those being developed to combat human immunodeficiency virus infection.

Adjuvants, Immunologic↗

Establishment of reverse genetics systems for Colorado tick fever virus.

The Colorado tick fever virus (CTFV), which has 12-segmented double-stranded RNA genomes, is a pathogenic arbovirus that causes severe diseases in humans. However, little progress has been made in the analysis of replication mechanisms and pathogenicity. This virological constraint is due to the absence of a reverse genetics system for CTFV; therefore, we aimed to establish the system. Initially, the efficacy of CTFV replication was investigated in various cell lines. CTFV was found to grow in many cell types derived from different hosts and organs. Subsequently, BHK-T7 cells stably expressing T7 RNA polymerase were transfected with plasmids encoding each of the 12 CTFV gene segments, expression plasmids encoding all CTFV proteins, and a vaccinia virus RNA-capping enzyme. Following transfection, the cells were co-cultured with Vero or HeLa cells. Using this system, we rescued monoreassortants and recombinant viruses harboring peptide-tagged viral proteins. Furthermore, an improved system using Expi293F cells expressing T7 RNA polymerase was established, which enabled the generation of recombinant reporter CTFVs. In conclusion, these reverse genetics systems for CTFV will greatly contribute to the understanding of viral replication mechanisms, pathogenesis, and transmission, ultimately facilitating the development of rational treatments and candidate vaccines.

Animals↗

Targeting ticks for control of selected hemoparasitic diseases of cattle.

Development in and transmission of hemoparasites by tick vectors are phenomena closely synchronized with the tick feeding cycle. In all known life cycles, initial infection of tick tissues occurs in midgut epithelial cells and transmission is effected as ticks feed after parasites have developed and multiplied in salivary glands. Many factors reviewed affect development and transmission of hemoparasites by ticks including age of ticks, artificial temperature, climate and/or season, tick stage or sex, hemoparasite variation, concurrent infection of ticks with other pathogens, host cell susceptibility, transovarial transmission, effect of hemoparasites on tick biology, and the effect of infecting parasitemia level in cattle on infection rates in ticks. Four hemoparasites of cattle, Anaplasma marginale, Cowdria ruminantium, Theileria parva, and Babesia spp., are all dependent on ticks for biological transmission. Babesia is transmitted transovarially whereas the other three are transmitted transstadially. Mechanical transfer of infective blood via fomites and mouthparts of biting arthropods is also a major means of transmission for Anaplasma marginale but not of the others. Potential control methods for hemoparasites that target parasites as they are developing in their respective tick hosts include tick control, vaccines (against ticks and parasites), and drugs (against ticks and parasites). Successful application of control strategies will be dependent upon thorough understanding of parasite developmental cycles, biology of the tick vectors and the immune response of cattle to ticks and to hemoparasites. The most effective control measures will be those that are targeted against both ticks and the hemoparasites they vector.

Animals↗

Plaque production by arboviruses in Singh's Aedes albopictus cells.

We report plaquing tests of 124 virus strains, mostly arboviruses of 21 serological groups, in Singh's line of Aedes albopictus cells. Thirty of these plaqued: all were arboviruses of six groups and were known or presumed to be mosquito borne. Failing to plaque were 86 strains of arboviruses, mostly tick borne, two strains of insect pathogens, and six animal viruses not classified as arboviruses. Among mosquito-borne agents, plaquing ability appeared related to serological classification. California group and most A-group viruses failed to plaque, but nearly all members of B and Bunyamwera groups readily plaqued. Within serological group B, 14 of 16 mosquito-borne agents plaqued, but none of 13 tick-borne or vector-unassociated viruses did so. Some implications of these results for recognition and classification of arboviruses are discussed.

Aedes↗

DNA characterization of the spirochete that causes Lyme disease.

Lyme disease, a tick-borne disease long recognized in Europe but only recently recognized in the United States, was shown in 1982-1983 to be caused by a spirochete, the Lyme disease spirochete. Whether one or more species of the spirochete exists is unknown, as is its taxonomic status. To answer these questions, we determined (i) the DNA base (guanidine-plus-cytosine) content for five strains; (ii) the DNA relatedness of 10 strains from Europe or the United States (isolated from ticks, humans, and a mouse) by DNA hybridization (hydroxyapatite assay at 50 and 65 degrees C); and (iii) the DNA relatedness to other pathogenic spirochetes. The guanine-plus-cytosine content of the Lyme disease spirochete strains was 27.5 to 29.0 mol%, most similar to those of Borrelia hermsii (30.6 mol%) and Treponema hyodysenteriae (25.6 mol%) among the other spirochetes tested. DNA hybridization studies with 32P-labeled DNA from Lyme disease spirochete strain TLO-005, a human blood isolate, revealed divergence (unpaired bases) within related nucleotide sequences of only 0.0 to 1.0% for all nine Lyme disease spirochete strains tested for relatedness to TLO-005. Relatedness values of seven strains to TLO-005 were 58 to 98% (mean, 71%) in 50 degrees C reactions and 50 to 93% (mean, 69%) in 65 degrees C reactions. Two other strains, from which very low yields of DNA were obtained, showed less relatedness (36 to 50 degrees C, 38 to 47% at 65 degrees C). These were nonetheless considered to belong to the same species because of the low amount of divergence in the sequences related to TLO-005 and the absence of decreased relatedness in reactions done at 65 degrees Celsius compared with those done at 50 degrees Celsius. DNA from strain TLO-005 showed relatedness of 1% to DNAs of two leptospires and 16% relatedness to DNA from T. hyodysenteriae. B. hermsii DNA was 30 to 40% related to three Lyme disease spirochete strains in 50 degrees Celsius reactions. Divergence in these reactions was 16.5 to 18.5%, and relatedness in 65 degree Celsius reactions was 8 to 10%. On the bases of phenotypic similarity, guanine-plus-cytosine content, and DNA relatedness to B. hermsii, we conclude the Lyme disease spirochete is single previously undescribed species which belongs in the genus Borrelia.

Animals↗

Identification of the tick-borne relapsing fever spirochete Borrelia hermsii by using a species-specific monoclonal antibody.

Borrelia hermsii causes a relapsing fever in humans and is one of several species of tick-borne spirochetes known to occur in the western United States. Spirochetes observed in the peripheral blood of patients acutely ill have been presumptively identified in the past by the geographic location of exposure and the probable species of tick vector. We describe a monoclonal antibody (H9826) that bound to the flagellar protein of B. hermsii but not to those of any of the other species tested, which included B. parkeri, B. turicatae, B. coriaceae, B. anserina, B. burgdorferi, and Leptospira interrogans serovar ballum. This antibody bound efficiently to B. hermsii in an indirect immunofluorescence assay and was used to rapidly detect and identify this spirochete in the peripheral blood of experimentally infected mice and in the central ganglia of Ornithodoros hermsi ticks. H9826 can rapidly confirm the identification of B. hermsii to increase our understanding concerning the geographic distribution, vector specificity, and epidemiological significance of this zoonotic human pathogen.

Antibodies, Bacterial↗

Serotypes of tick-borne spotted fever group rickettsiae from western California.

A rickettsial survey of ixodid ticks known to bite man was conducted in 1979 in four coastal counties of California to obtain isolates from tick species that might be involved in the transmission of spotted fever-like illnesses, and to examine serologic characteristics of the rickettsiae relative to defined members of the spotted fever group (SFG). One hundred and seventy (19.4%) of 877 ticks comprising three species were shown by hemolymph test to harbor rickettsia-like organisms. A total of 85 SFG rickettsia isolates was obtained by Vero-cell culture; 82 were from Dermacentor occidentalis, two were from D. variabilis, and one was from Ixodes pacificus. As determined by microimmunofluorescence, the isolates comprised four distinct serotypes. Two serotypes were obtained only from D. occidentalis, and one each only from D. variabilis and I. pacificus, respectively. Most D. occidentalis isolates possessed the serologic characteristics of Rickettsia rhipicephali, but there were similar to yet distinguishable from, R. rickettsii and are members of an unclassified serotype referred to as 364D. The two isolates from D. variabilis resembled the unclassified 369C serotype previously shown to be associated with this species and D. andersoni elsewhere in the United States. The I. pacificus isolate was similar to strains of the unclassified Tillamook serotype isolated from this tick in several localities in western Oregon. Representative strains of the four serotypes could also be distinguished on the basis of pathogenicity for Vero cells, chick embryos, guinea pigs, and/or meadow voles. The significance of these findings relative to occurrence of tick-associated illnesses in western California is briefly discussed.

California↗

Variability of Powassan virus cultured in tissue explants and organism of Hyalomma anatolicum ticks.

The variability of Powassan virus was studied during successive passages in Hyalomma anatolicum ticks or prolonged reproduction in their tissue explants. It had been shown that in the course of tick passages and during reproduction in the explants, pathogenicity of the virus in respect to causing acute disease in mice after peripheral inoculation was decreased, while virus ability to cause death after intracerebral (i.c.) inoculation remained unchanged. In mice infected with the original strain P-40 of Powassan virus damaging effect of the immune response prevailed, while in mice infected with the strains passaged for a long time in ticks (strain P-40Hat) or in tick tissue explants (strain P-40Haex), the protective effect of the immune response was prominent.

Animals↗

[Tick-borne viruses in tropical area (author's transl)].

Arboviruses, transmitted by ticks and pathogenic to man in tropical areas, are discussed. They are considered according to, first the faunal region in which they can be found, then the type of syndrome they cause. The greater part is of little medical importance, some are noxious but of a rare occurrence (Thogoto, Wanowrie, Langat) or geographically restrained (Kyasanur forest disease), so the Crimean-Congo hemorrhagic fever is the only infection to show a notorius importance. As severe in the tropics of the oriental faunal region (Pakistan, India) as it is Europe, it seems to be of little importance in the afrotropical faunal region. Nevertheless, its severity remains to be ascertained by the means of extensive serological surveys and by taking in account this virus in presence of every hemorrhagic syndrome.

Animals↗

[Tick-borne arboviruses associated with marine birds; a general review (author's transl)].

About forty different arboviruses have been isolated from ticks associated with marine birds along all the coast of the world. The majority of these viruses belongs to 8 serogroups: Hughes, B, Kemerovo, Quaranfil, Nyamanini, Sakhalin, Unkuniemi and Upolu, while some others are non-grouped arboviruses. Two kinds of specialized ticks chiefly harbour these arboviruses: 1. The ticks of Ornitbodoros capensis complex from tropical, subtropical and temperate zones - 2. The ticks Ixodes uriae (= I. putus) and Ixodes signatus from subpolar, arctic and antarcitc zones and the surrounding temperate areas. All these ticks may attack humans, and severe pruritus and/or undifferenciated febrile illness may result. From a pathological point of view, some of these arboviruses, i.e. Soldado, Aride, Saumarez Reef, act as pathogens for marine birds inducing nest desertion and death or abnormal development of newly batched chicks. On the other hand, antibodies against other viruses, with well known or suspected epidemic potency, such as West Nile, Tyuleniy, Okhotskiy viruses, are currently found in human beings in certain coastal areas. These epidemiological data deserve further attention from concerned Health autorities.

Animals↗

[Rickettsia slovaca: an agent of the group of exanthematous fevers, in Portugal].

BACKGROUND: Identification of Rickettsia isolated from Dermacentor marginatus ticks from Portugal. METHODS: Using recently developed techniques (shell-vial isolation, PCR/RFLP genetical methods, SDS-PAGE), serological studies and animal induced pathology to isolate and characterize tick-borne rickettsiae. RESULTS: A total of 632 adults D. marginatus ticks were captured. From the collected ticks, we isolated 18 Rickettsia strains and the results confirm that the isolates from the haemolymph were identical to R. slovaca reference strain. CONCLUSIONS: Dermacentor marginatus ticks infected with Rickettsia slovaca were found in Portugal. This occurrence extended the known, geographical area for this microorganism. It is also a factor that could alter the seroprevalence rates of butonneuse fever. The pathogenicity of this Rickettsia should be evaluated to determine its responsibility for diseases other than butonneuse fever.

Animals↗