In 1937, Derrick [1] described a disease in 20 out of 800 workers of a meat factory in Brisbane, Queensland, Australia. Since this disease was unknown, he called it query fever (Q fever). The pathogen was isolated from the blood and urine of the patients in Australia by Burnet and Freeman and was called Rickettsia (R. burnetii) [2]. At the same time, the pathogen was isolated from ticks in Montana, USA, by Davis and Cox [3] who called it Rickettsia diaporica. Later, it was renamed into Coxiella burnetii since one did not wish to fail to acknowledge either research group. In Germany, the infection was known in the army as ‘Balkan flu’ during World War II [4]. C. burnetii infection occurs worldwide. It is a zoonosis transmitted to humans especially by infected cattle, sheep and goats, but also by cats, dogs, rabbits, ducks, the faeces of ticks and dust. The pathogen causes acute or chronic infection and is acquired by contact or through airborne dissemination or inhalation of contaminated dust or consumption of unpasteurised milk and milk products [5, 6]. More than 20 different genomic C. burnetii strains have been described. As has been established using pulse-field gel electrophoresis, European strains differ from North American ones [7], but there are also regional strains, and differences exist between e.g. regional groups in Germany and in Russia. The genome length of the bacterium is between 1.5 and 2.1 million base pairs (bp).

The human immunodeficiency virus (HIV) is a member of the group of Lentivirinae in the family of Retroviridae, while the human T-cell leukemia virus (HTLV) is an oncovirus [1]. HIV is subdivided into types 1 and 2 (HIV-1 and HIV-2) on the basis of genetic properties and differences in antigens. The immunodeficiency viruses in monkeys are known as simian immunodeficiency viruses (SIV). The currently available epidemiological and molecular biological data indicate that HIV was transmitted from African primates to humans in the mid20th century [2].

1.1 Characteristics of HCMV Together with animal cytomegalovirus, human cytomegalovirus (HCMV), also referred to in recent literature as human herpesvirus 5 (HHV-5), belongs to the Herpesviridae family, subfamily Betaherpesvirinae, genus Cytomegalovirus. The name is derived from the fact that it causes enlargement of the infected cell (cytomegaly) and induces characteristic inclusion bodies. The HCMV genome consists of a doublestranded DNA with approximately 230,000 base pairs. The genome is enclosed by an icosahedral capsid (100–110 nm diameter, 162 capsomers). Between the capsid and the virus envelope is a protein layer known as the tegument. The virus envelope is derived from cell membranes. At least eight different viral glycoproteins are embedded in the lipid bilayer. The mature viral particle has a diameter of 150–200 nm (fig. 1). Like all herpesviruses, HCMV is sensitive to low pH, lipiddissolving agents and heat. HCMV has a half-life of approximately 60 min at 37 °C and is relatively unstable at –20 °C. It needs to be stored at at least –70 °C in order to maintain its infectivity. A distinction is made with herpesviruses between a) the lytic infection cycle and b) latency which leads to life-long infection of the organism. Characteristics of the beta-herpesviruses are their high level of host specificity, their slow replication cycle and the spread of infection from cell to cell in the cell culture even in the presence of neutralising antibodies. The lytic infection of cells can be monitored using protein expression patterns and the replication of nucleic acid. The immediate early (IE) proteins are responsible for the regulation of the early (E) proteins and also for that of the late (L) proteins. After adsorption of the virus onto the target cell with the aid of viral glycoproteins, the virus envelope fuses with the cell membrane, the capsid is released into the cell and is transported to the nucleus where the genome is released. Transcription of the IE proteins then takes place in the cell nucleus with the aid of the RNA polymerase II of the host cell. Tegument proteins of the infecting virus particle act as transactivators for the IE genes. The IE proteins regulate the following stages of viral replication and are also involved with cell regulation, including the expression and recognition of the HLA antigens (class I MHC proteins). IE proteins (in particular the phosphoprotein pp65) can be used as early markers of the virus infection in cell cultures. The E proteins include the HCMV-coded DNA polymerase the activity of which can be specifically inhibited with antiviral agents which have to be phosphorylated by viral nucleotide kinases. The synthesis of the structural proteins (L proteins) is regulated via the E proteins. The viral capsids are formed in the cell nucleus; export and enveloping of the viruses take place on the inner nuclear membrane (possibly also on other cell membranes). HCMV shows pronounced cellular association. Studies with monoclonal antibodies point to differences between viral strains and isolates. The most recent studies using primary isolates and the corresponding sera from patients show that strain-specific neutralising antibodies are formed. Whether the variability of antigens is the main cause here as with other virus families or whether there are serological subtypes has not yet been clarified [1].

ABSTRACT An infectious parvovirus B19 (B19V) genotype 2 variant was identified as a high-titer contaminant in a human plasma donation. Genome analysis revealed a 138-bp insertion within the p6 promoter. The inserted sequence was represented by an additional 30 bp from the end of the inverted terminal repeat adjacent to a 108-bp element found also, in inverted orientation, at the extreme right end of the unique sequence of the genome. However, despite the profound variations in the promoter region, the pattern of gene expression and DNA replication did not differ between genotype 1 and genotype 2 in permissive erythroid KU812Ep6 cells. Capsid proteins of both genotypes differ in their amino acid sequences. However, equivalent kinetics of virus inactivation at 56°C or pH 4 indicated a comparable physicochemical stability of virus capsids. Sera from six individuals infected by B19V genotype 1 were investigated on cross-neutralization of B19V genotype 2 in vitro. Similar neutralization of both B19V genotypes was observed in sera from three individuals, while the sera from three other individuals showed weaker cross-neutralization for genotype 2. In conclusion, the in vitro replication characteristics and physical stability of B19V capsids are very similar between human parvovirus B19 genotypes 1 and 2, and cross-neutralization indicates a close antigenic relation of genotypes 1 and 2.

‘Arboviruses’ (arthropod-borne viruses) is a collective term for viruses that replicate in both arthropods, such as mosquitoes or ticks, and vertebrates (birds, mammals). These viruses may be transmitted to vertebrates by arthropods via a bite when taking a blood meal. Arboviruses currently include approximately 400 representatives of different families or genera of viruses. Table 1 summarises the pathogens of greatest importance to humans, the vectors involved, the virus reservoirs and the geographical distribution of the diseases. Not every genus of a virus family includes pathogens of human diseases that are transmitted by arthropods. For example, only 3 out of the total of 6 genera in the family Bunyaviridae contain viral species that are transmissible to humans by arthropods. Hantaviruses, on the other hand, which belong to the Hantavirus genus of the Bunyaviridae family, are transmitted to humans not by arthropods but through human contact with excreta of the natural hosts, mice and rats. A high level of safety precautions is necessary when working with arboviruses as these pathogens are classified predominantly in risk groups 3 or 4 [1, 2].

The coding sequences (open reading frame, ORF) of the genome are flanked at both sides by noncoding sequences (noncoding region 5'-NCR and 3'-NCR). The 5'-NCR, as in HCV, contains an internal ribosomal entry site (IRES) for protein synthesis. In all isolates currently known the core sequence (C) is lacking or shortened with respect to the HCV core sequence. As in HCV one observes two virus envelope proteins (E1 and E2). The viral proteases are encoded by the genes for nonstructural proteins NS2 and NS3. NS3, as in all flaviviruses, also contains a helicase. For the genome regions NS4 and NS5A the function of the encoded proteins has not yet been elucidated. The NS5B gene encodes the viral replicase (RNA-dependent RNA polymerase).

Jaagsiekte retrovirus (JSRV) causes ovine pulmonary adenomatosis (OPA) that resembles bronchioloalveolar carcinoma (BAC) in humans. To test the possible role of JSRV in human diseases, DNA specimens from 103 individuals either healthy or suffering from lung carcinomas were analyzed for JSRV sequences. orf-x sequences were detected in 19 of 64 samples and gag-prt sequences in 4 of 38 samples, predominantly in individuals from Africa. Sequences obtained from orf-x amplimers varied in-between each other and differed from control endogenous ovine JSRV sequence. No association with lung cancer was found. This is the first report of JSRV-like sequences detected in humans.

Phylogenetically new insertions of repetitive sequences may contribute to genome evolution by altering the function of preexisting proteins. One example is the SVA sequence, which forms the C-terminal coding exon of the human leptin receptor isoform 219.1. Here, we report that the SVA insertion into the LEPR locus has occurred after divergence of humans and chimpanzees. The SVA element was inserted into a Hal-1/LINE element present in all monkeys and apes tested. Structural features point toward an integration event that was mediated by the L1 protein machinery acting in trans. Thus, our findings add evidence to the hypothesis that retrotransposition events are a driving force in genomic evolution and that the presence or absence of specific retroelements are one distinguishing feature that separates humans from chimpanzees.

Background: Human parvovirus B19 (B19) DNA can be frequently detected in plasma-derived coagulation factor concentrates. The production of some clotting factor products includes heat treatment steps for virus inactivation, but the effectiveness of such steps for B19 inactivation is unclear. Moreover, detailed transmission case reports including DNA sequence analysis and quantification of B19 DNA from contaminated heat-treated blood components have not been provided so far. Therefore, the correlation between B19 DNA in blood components and infectivity remains unclear.Study design and methods: Asymptomatic B19 infections of two patients with hemophilia A were detected by anti-B19 seroconversion after administration of B19-contaminated heat-treated clotting factors. The suitability of nucleic acid sequence analysis for confirmation of B19 transmission was investigated. Furthermore, the B19,DNA level in blood components was determined and the drug administration was reviewed to calculate the amount of inoculated B19 DNA.Results: Both B19 transmissions from clotting factor products could be confirmed by identical nucleic acid sequences of virus DNA from patients and blood components while sequences from unrelated controls could be differentiated. One patient received, for 4 days, a total of 180 mL vapor heat-treated prothrombin complex concentrate containing 8.6x10(6) genome equivalents per mL of B19 DNA. The other patient received 966 mL of low-contamination (4.0x10(3) genome equivalents/mL) dry heat-treated FVIII concentrate over a period of 52 days.Conclusion: B19 transmissions can be confirmed by nucleic acid sequencing. However, due to the low variability of the B19 genome, a large part of the B19 genome must be analyzed. The transmissions show that the applied heat treatment procedures were not sufficient to inactivate B19 completely.

A new problem on hepatitis for Indonesian is hepatitis-C virus (HCV). This infection is endemic, majority sub-clinic and progressive in chronic. Viral transmission is primarily via a parenteral route, while other routes are still in debate. Diagnostic approach should be focused on how this virus developed. KeyWords: hepatitis-C virus molecular biology Westem-blot-HCV blood transfusion epidemiology

A mixture of Tri-n-butyl phosphate (TNBP) and Polysorbate 80 (Tween 80) is often used for virus inactivation during the manufacture of medicinal products derived from human plasma. This procedure, known as solvent/detergent treatment, is of high effectiveness for inactivation of enveloped viruses. Tween 80 can be manufactured from bovine tallow or from vegetable material. As the bovine-derived Tween 80 is normally used for the solvent/detergent treatment, the question has been raised whether vegetable-derived Tween 80 can be applied as an alternative substance for the solvent/detergent treatment. Comparable inactivation studies were therefore performed using Vesicular Stomatitis Virus (VSV), Pseudorabiesvirus (PRV), Semliki Forest Virus (SFV) and Bovine Diarrhoea Virus (BVDV). In principle, no differences were observed in the effectiveness of the solvent/detergent treatment when bovine or vegetable-derived Tween 80 was used. The comparability in the efficiency of both detergents for virus inactivation was shown to be independent of solvent/detergent concentration, of temperature (16°C and 6°C vs. 27°C and 25°C) and protein concentration (10% and 5% human albumin). In summary, vegetable-derived Tween 80 is of the same effectiveness as bovine-derived Tween 80, when used for virus inactivation by the solvent/detergent treatment. Copyright 2002 The International Association for Biologicals. Published by Elsevier Science Ltd. All rights reserved.

BACKGROUND : It has been shown that HSA may be contaminated with parvovirus B19 (B19) DNA. However, the presence of B19 DNA does not necessarily indicate infectious virus. HSA is pasteurized at 60°C for 10 hours and it remains unclear whether this procedure inactivates B19. Studies with animal parvoviruses indicate considerable heat resistance at 60°C. However, due to the lack of a suitable cell culture system, the pasteurization process has not been investigated in the past. STUDY DESIGN AND METHODS : The recently described cell clone KU812Ep6 was used to establish a system for investigation of B19 inactivation during pasteurization. Virus‐infected cells were detected by immunofluorescent staining of viral capsid antigen and by RT‐PCR assay of virus‐specific capsid mRNA. RESULTS : B19 was inactivated after 10 minutes at 60°C for ≥4 log . In contrast, porcine parvovirus was widely resistant at 60°C. Inactivation of B19 was independent of the analyzed albumin products (5, 20, and 25% albumin from three manufacturers) and from the specific virus source used for the inactivation experiments. Degradation of B19 DNA by deoxyribonuclease I treatment after pasteurization indicated that the virus capsid is destroyed during heat treatment. CONCLUSION : Heat resistance of B19 markedly differs from heat resistance of animal parvoviruses. While animal parvoviruses widely withstand pasteurization of albumin, B19 was rapidly inactivated. These results confirm the safety of pasteurized albumin and are in line with its good clinical safety record with respect to B19 infection. However, conclusions regarding the safety of other blood‐derived medicinal products should not be derived from B19 inactivation in albumin, because different processes or different composition of product intermediates may significantly influence B19 stability during heat treatment.

Vox SanguinisVolume 81, Issue 3 p. 204-206 Human T-cell lymphocytotrophic virus prevalence in German blood donors and ‘at-risk’ groups M. Nübling Dr, M. Nübling DrSearch for more papers by this authorC. M. Nübling, C. M. Nübling Paul-Ehrlich-Institut (PEI), Abteilung für Virologie, Langen, GermanySearch for more papers by this authorE. Seifried, E. Seifried Institut für Transfusionsmedizin und Immunhämatologie, Blutspendedienst des Deutschen Roten Kreuzes gGmbH, Frankfurt, GermanySearch for more papers by this authorW. Weichert, W. Weichert Institut für Transfusionsmedizin und Immunhämatologie, Blutspendedienst des Deutschen Roten Kreuzes gGmbH, Frankfurt, GermanySearch for more papers by this authorJ. Löwer, J. Löwer Paul-Ehrlich-Institut (PEI), Abteilung für Virologie, Langen, GermanySearch for more papers by this author M. Nübling Dr, M. Nübling DrSearch for more papers by this authorC. M. Nübling, C. M. Nübling Paul-Ehrlich-Institut (PEI), Abteilung für Virologie, Langen, GermanySearch for more papers by this authorE. Seifried, E. Seifried Institut für Transfusionsmedizin und Immunhämatologie, Blutspendedienst des Deutschen Roten Kreuzes gGmbH, Frankfurt, GermanySearch for more papers by this authorW. Weichert, W. Weichert Institut für Transfusionsmedizin und Immunhämatologie, Blutspendedienst des Deutschen Roten Kreuzes gGmbH, Frankfurt, GermanySearch for more papers by this authorJ. Löwer, J. Löwer Paul-Ehrlich-Institut (PEI), Abteilung für Virologie, Langen, GermanySearch for more papers by this author First published: 28 July 2009 https://doi.org/10.1046/j.1423-0410.2001.00102.xCitations: 6Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume81, Issue3October 2001Pages 204-206 RelatedInformation

Zusammenfassung Viele Substanzen, die für die Herstellung von Arzneimitteln notwendig sind, werden aus Organen, Geweben oder Körperflüssigkeiten von Rindern gewonnen. Daher stellt sich die Frage nach der BSE-Sicherheit auch von immunbiologischen Arzneimitteln wie Impfstoffen, Allergenpräparaten und Blutprodukten. Um eine vernünftige Risikoeinschätzung vornehmen zu können, müssen drei Aspekte berücksichtigt werden: Welche Art von Rinderbestandteilen eingesetzt werden (Rindermaterialien oder Rinderprodukte), ob diese als Ausgangsstoff für ein Arzneimittel dienen bzw. im Fertigarzneimittel enthalten sind oder ob sie als Hilfssubstanzen zur Herstellung benötigt, aber im Laufe der Herstellung wieder entfernt werden, in welche Risikokategorien die Organe und Gewebe eingestuft werden, von denen die verwendeten Rindermaterialien stammen oder aus denen die Rinderprodukte gewonnen werden. Nach diesen Kriterien werden für die Produktgruppen Impfstoffe, Allergenpräparate und Blutprodukte jeweils vier Fälle durchgespielt, die der Frage nachgehen, in welcher Form Substanzen vom Rind eingesetzt werden und ob dadurch ein BSE-Risiko von diesen Produkten ausgeht. Anhand dieser jeweils vier Fälle wird deutlich, dass von immunbiologischen Arzneimitteln, die in Deutschland in Verkehr sind, keine BSE-Gefahr ausgeht.

The principle of live attenuated influenza vaccines has been known for many decades. However, the pharmaceutical and clinical development according to current regulations, of modern live influenza vaccines based on cold adapted influenza viruses (CAIV) started only recently and these vaccines will most probably become an alternative within the next couple of years to licensed inactivated influenza vaccines that have been used routinely since the early 1940s. In contrast to contemporary trivalent inactivated influenza vaccines, which are administered intramuscularly, trivalent CAIV-based vaccines will be administered intranasally as a spray. Quality, safety and efficacy aspects related to CAIV vaccines as well as possible risks linked to the widespread use of these vaccines will be discussed in the following overview and compared to established influenza vaccines. Moreover, issues of practicality of CAIV vaccines focusing on the necessity of an annual update of influenza vaccines are addressed.

ABSTRACT It was recently reported that the human endogenous retrovirus HTDV/HERV-K encodes the regulatory protein Rec (formerly designated Corf), which is functionally equivalent to the nuclear export adapter proteins Rev of human immunodeficiency virus and Rex of human T-cell leukemia virus. We have demonstrated that the Rec protein interacts with a characteristic 429-nucleotide RNA element, the Rec-responsive element (RcRE), present in the 3′ long terminal repeat of HTDV/HERV-K transcripts. In analogy to the Rev and Rex proteins, which have distinct RNA binding sites in their responsive elements, we have proposed that Rec may also have a defined binding site in the RcRE. In this report, we demonstrate that not every HTDV/HERV-K copy present in the human genome contains an active RcRE, and we characterize mutations that abrogate Rec function. In addition, we demonstrate that Rec function requires binding to a complex, folded RNA structure rather than binding to a discrete specific binding site, in contrast to Rev and Rex and their homologous responsive elements. We define four stem-loop structures in the RcRE that are essential for Rec function. Finally, we demonstrate that both Rev and Rex can mediate nuclear export through the RcRE but that their binding sites are different from each other and from that of Rec.