Origin of human T-lymphotropic virus type 1 in rural Côte d'Ivoire.

Simian T-lymphotropic virus type 1 (STLV-1) strains occasionally infect humans. However, the frequency of such infections is unknown. We show that direct transmission of STLV-1 from nonhuman primates to humans may be responsible for a substantial proportion of human T-lymphotropic virus type 1 infections in rural Côte d'Ivoire, where primate hunting is common.

Pandemic human viruses cause decline of endangered great apes.

Commercial hunting and habitat loss are major drivers of the rapid decline of great apes [1]. Ecotourism and research have been widely promoted as a means of providing alternative value for apes and their habitats [2]. However, close contact between humans and habituated apes during ape tourism and research has raised concerns that disease transmission risks might outweigh benefits [3-7]. To date only bacterial and parasitic infections of typically low virulence have been shown to move from humans to wild apes [8, 9]. Here, we present the first direct evidence of virus transmission from humans to wild apes. Tissue samples from habituated chimpanzees that died during three respiratory-disease outbreaks at our research site, Côte d'Ivoire, contained two common human paramyxoviruses. Viral strains sampled from chimpanzees were closely related to strains circulating in contemporaneous, worldwide human epidemics. Twenty-four years of mortality data from observed chimpanzees reveal that such respiratory outbreaks could have a long history. In contrast, survey data show that research presence has had a strong positive effect in suppressing poaching around the research site. These observations illustrate the challenge of maximizing the benefit of research and tourism to great apes while minimizing the negative side effects.

A new flavivirus and a new vector: characterization of a novel flavivirus isolated from uranotaenia mosquitoes from a tropical rain forest.

A novel flavivirus was isolated from Uranotaenia mashonaensis, a mosquito genus not previously known to harbor flaviviruses. Mosquitoes were caught in the primary rain forest of the Taï National Park, Côte d'Ivoire. The novel virus, termed nounané virus (NOUV), seemed to grow only on C6/36 insect cells and not on vertebrate cells. Typical enveloped flavivirus-like particles of 60 to 65 nm in diameter were detected by electron microscopy in the cell culture supernatant of infected cells. The full genome was sequenced, and potential cleavage and glycosylation sites and cysteine residues were identified, suggesting that the processing of the NOUV polyprotein is similar to that of other flaviviruses. Phylogenetic analyses of the whole polyprotein and the NS3 protein showed that the virus forms a distinct cluster within the clade of mosquito-borne flaviviruses. Only a distant relationship to other known flaviviruses was found, indicating that NOUV is a novel lineage within the Flaviviridae.

High prevalence of porcine Hokovirus in German wild boar populations.

Porcine Hokovirus (PHoV) was recently discovered in Hong Kong. This new Parvovirus of pigs is closely related to the human Parvoviruses 4 and 5 (PARV4/5) and bovine Hokovirus (BHoV). So far, nothing is known about the presence and prevalence of PHoV in regions of the world other than Hong Kong. A study was initiated to investigate PHoV in German wild boars from five different geographical regions, using a newly established quantitative real-time PCR assay. Analysis of collected liver and serum samples revealed high overall prevalence (32.7%; 51/156) of PHoV in wild boars. The prevalence differed between the regions and increased with age. Two near full-length genomes and a large fragment for three additional isolates from different regions were sequenced and used for phylogenetic analysis. The German PHoV sequences from wild boars showed a close relationship with sequences of isolates from Hong Kong.

Anthrax kills wild chimpanzees in a tropical rainforest.

Infectious disease has joined habitat loss and hunting as threats to the survival of the remaining wild populations of great apes. Nevertheless, relatively little is known about the causative agents. We investigated an unusually high number of sudden deaths observed over nine months in three communities of wild chimpanzees (Pan troglodytes verus) in the Taï National Park, Ivory Coast. Here we report combined pathological, cytological and molecular investigations that identified Bacillus anthracis as the cause of death for at least six individuals. We show that anthrax can be found in wild non-human primates living in a tropical rainforest, a habitat not previously known to harbour B. anthracis. Anthrax is an acute disease that infects ruminants, but other mammals, including humans, can be infected through contacting or inhaling high doses of spores or by consuming meat from infected animals. Respiratory and gastrointestinal anthrax are characterized by rapid onset, fever, septicaemia and a high fatality rate without early antibiotic treatment. Our results suggest that epidemic diseases represent substantial threats to wild ape populations, and through bushmeat consumption also pose a hazard to human health.

Interspecies transmission of simian foamy virus in a natural predator-prey system.

Simian foamy viruses (SFV) are ancient retroviruses of primates and have coevolved with their host species for as many as 30 million years. Although humans are not naturally infected with foamy virus, infection is occasionally acquired through interspecies transmission from nonhuman primates. We show that interspecies transmissions occur in a natural hunter-prey system, i.e., between wild chimpanzees and colobus monkeys, both of which harbor their own species-specific strains of SFV. Chimpanzees infected with chimpanzee SFV strains were shown to be coinfected with SFV from colobus monkeys, indicating that apes are susceptible to SFV superinfection, including highly divergent strains from other primate species.

Decontamination of Personal Protective Equipment.

Exploratory field analyses of the inactivation capacity of disinfectants on contaminated personal protective equipment (PPE) are required to select a suitable surrogate for biohazardous agents like spores of Bacillus anthracis. The objectives of our study were (1) the determination of an appropriate surrogate for the inactivation of spores of B. anthracis with peracetic acid (PAA), and (2) application of optimized inactivation conditions for an effective decontamination of PPE with PAA under field conditions. For inactivation studies, B. anthracis spores from different strains and B. thuringiensis spores were fixed by air drying on carriers prepared from PPE fabric. Time and concentration studies with PAA-based disinfectants revealed that the spores of the B. thuringiensis strain DSM 350 showed an inactivation profile comparable to that of the spores of the B. anthracis strain with the highest stability, implying that B. thuringiensis can serve as an appropriate surrogate. Rapid (3 to 5 minutes) and effective surface decontamination was achieved with 2% PAA/0.2% surfactant. In field studies, PPE contaminated with spores of B. thuringiensis was treated with the disinfectant. Optimizing the decontamination technique revealed that spraying in combination with brushing was effective within 5 minutes of exposure.

Orthopoxvirus detection in environmental specimens during suspected bioterror attacks: inhibitory influences of common household products.

After terrorists attacked the United States in 2001, the appearance of letters and other objects containing powdery substances with unknown potentials for biological threat focused attention on the speed, sensitivity, and reliability of diagnostic methods. This study summarizes the abilities and limitations of real-time PCR, electron microscopy (EM), and virus isolation when used to detect potential bioweapons. In particular, we investigated the inhibitory influences of different common household products present in environmental specimens on PCR yield, EM detection, and virus isolation. We used vaccinia virus as a model for orthopoxviruses by spiking it into specimens. In the second part of the study, we describe modifications of diagnostic methods to overcome inhibitory effects. A variety of PCR amplification enhancers, DNA extraction protocols, and applications of internal controls were evaluated to improve diagnostic simplicity, speed, and reliability. As a result, we strongly recommend using at least two different frontline techniques in parallel, e.g., EM and PCR. A positive result obtained by any one of these techniques should be followed by a biological method to confirm the putative diagnosis. Confirmatory methods include virus isolation followed by an agent-specific immunofluorescence assay to confirm the presence of replication-competent particles.

Lack of detectable West Nile virus RNA in brains and kidneys of dogs and cats with immunohistological precipitates using virus-specific antibodies.

Five dogs and four cats from Germany suffering from encephalitis revealed positive immunoreactivity using two West Nile virus (WNV) specific monoclonal antibodies in brain and in kidney. However, WNV-infection could not be confirmed by additional PCR analyses. This study indicated that positive immunoreactivity for WNV in dogs and cats must be interpreted cautiously and should be confirmed by a second virus-specific technique.

Cowpox infection transmitted from a domestic cat.

A 21-year-old immunocompetent woman developed a cowpox infec-tion,while working as a veterinarian's assistant in a rural area. She had never received vaccinia immunization and came in contact with a fatally-infected house cat. Under symptomatic treatment, the infection remained localized to one cheek and cleared over 3 weeks with substantial dermal-subcutaneous tissue destruction. Orthopoxvirus detection by PCR is a modern diagnostic standard, in addition to identification by negative-contrast electron microscopy. A promising therapeutic option is cidofovir, but this virostatic drug is not yet approved for the treatment of cowpox.

One-step selection of Vaccinia virus-binding DNA aptamers by MonoLEX.

BACKGROUND: As a new class of therapeutic and diagnostic reagents, more than fifteen years ago RNA and DNA aptamers were identified as binding molecules to numerous small compounds, proteins and rarely even to complete pathogen particles. Most aptamers were isolated from complex libraries of synthetic nucleic acids by a process termed SELEX based on several selection and amplification steps. Here we report the application of a new one-step selection method (MonoLEX) to acquire high-affinity DNA aptamers binding Vaccinia virus used as a model organism for complex target structures. RESULTS: The selection against complete Vaccinia virus particles resulted in a 64-base DNA aptamer specifically binding to orthopoxviruses as validated by dot blot analysis, Surface Plasmon Resonance, Fluorescence Correlation Spectroscopy and real-time PCR, following an aptamer blotting assay. The same oligonucleotide showed the ability to inhibit in vitro infection of Vaccinia virus and other orthopoxviruses in a concentration-dependent manner. CONCLUSION: The MonoLEX method is a straightforward procedure as demonstrated here for the identification of a high-affinity DNA aptamer binding Vaccinia virus. MonoLEX comprises a single affinity chromatography step, followed by subsequent physical segmentation of the affinity resin and a single final PCR amplification step of bound aptamers. Therefore, this procedure improves the selection of high affinity aptamers by reducing the competition between aptamers of different affinities during the PCR step, indicating an advantage for the single-round MonoLEX method.

Viremia in human Cowpox virus infection.

BACKGROUND: Several poxviruses can infect humans and cause diseases of varying severity. Besides the eradicated Variola virus that induced high mortality rates, numerous further human pathogenic orthopoxviruses are potentially fatal but generally cause less severe infections. While infection-related viremia was described for Variola virus and seems to be rare for Monkeypox virus, it is still debated for Vaccinia virus. So far, viremia in Cowpox virus-infected humans has not been reported. OBJECTIVES: To estimate the potential risk of Cowpox virus to disseminate and develop severe infections, two Cowpox virus patients were examined for viremia. STUDY DESIGN: Whole blood, serum and fluid from virus-induced lesions were analyzed by serology or quantitative real-time PCR. RESULTS: Real-time PCR and sequence analysis of the hemagglutinin gene confirmed Cowpox virus in the lesions of both patients. Serology performed on serum obtained at the same time as the lesion specimens demonstrated orthopoxvirus-specific IgG and IgM antibodies, indicating a recent orthopoxvirus infection. In addition, Cowpox virus DNA was detectable in whole blood, but not in serum, as late as week 4 post-infection. CONCLUSIONS: In contrast to observations following vaccination with Vaccinia virus, DNAemia in patients with localized symptoms of a Cowpox virus infection does not seem to be a rare event. However, its relevance for Cowpox virus pathogenicity has to be elucidated.

Serologic evidence of West Nile virus infections in wild birds captured in Germany.

To assess the risk of acquiring a West Nile virus (WNV) infection in Germany, we investigated samples from migrating and from resident birds. Because of their stay in or migration through WNV-endemic regions, these birds are at risk to become infected with WNV. Blood samples from 3,399 birds, representing 87 bird species, were collected in Germany in 2000 and in 2002-2005. Overall, 53 birds belonging to 5 species had WNV-neutralizing antibodies. Fifty-nine birds belonging to 9 species were reactive by WNV immunofluorescence assay, and 8 birds had neutralizing antibodies against Usutu virus. Because of maternal antibody transfer via egg yolk, WNV-antibody titers in white stork nestlings were generally lower than those in adults. Despite a relatively high percentage of stork nestlings with antibodies, no viral genomes were detectable by polymerase chain reaction. In Germany, the prevalence of antibodies to WNV in migrating birds wintering in Africa or southern Europe is comparatively low.

Detection of West Nile virus lineages 1 and 2 by real-time PCR.

West Nile virus (WNV) is a Flavivirus attracting worldwide attention because it has spread rapidly across the Americas since its first appearance in New York City in 1999. Several PCR-based diagnostic methods have been developed for the detection of WNV. The focus of these assays has been WNV lineage 1 which can be found worldwide, while lineage 2 viruses were thought to be endemic only in some regions of Africa. However, both lineages may be imported from Africa to Europe by migrating birds. In order to determine the incidence of WNV in Germany, a real-time-based PCR assay was developed, targeting a conserved region of WNV lineages 1 and 2. This assay is a suitable tool for the diagnosis of WNV and for surveillance studies.

Failure to detect Borna disease virus antigen and RNA in human blood.

BACKGROUND: Borna disease virus (BDV) is the etiological agent of a rare progressive meningoencephalitis that affects mostly horses and sheep. There is an unresolved debate whether also humans are susceptible to infection with BDV and if so, whether this might be associated with neuropsychiatric diseases. One recent key publication employing an ELISA-based sandwich assay reported prevalences of BDV-specific circulating immune complexes in human blood as high as 30% in the normal population and up to 100% in psychiatric patients [Bode L, Reckwald P, Severus WE, Stoyloff R, Ferszt R, Dietrich DE, et al. Borna disease virus-specific circulating immune complexes, antigenemia, and free antibodies--the key marker triplet determining infection and prevailing in severe mood disorders. Mol Psychiatry 2001;6(4):481-91]. However, this report did not examine for the physical presence of BDV antigens in human blood, and therefore, these seemingly high prevalences may not reflect Borna virus-specific signals. OBJECTIVES: We attempted to correlate string plasma signals in the particular sandwich ELISA system with the presence of BDV antigens. STUDY DESIGN: Four preselected plasma samples with high reactivity in the described assay were analysed by immunoaffinity purification and highly sensitive real-time RT-PCR. RESULTS: Neither method did provide any evidence for the presence of viral proteins or nucleic acids. CONCLUSIONS: Our findings argue against the concept that the described sandwich ELISA reliably detects BDV-specific antigens in human blood, therefore do not support the hypothesis that BDV is a pathogen of humans.