In order to generate recombinant bovine respiratory syncytial virus (BRSV), the genome of BRSV strain A51908, variant ATue51908, was cloned as cDNA. We provide here the sequence of the BRSV genome ends and of the entire L gene. This completes the sequence of the BRSV genome, which comprises a total of 15,140 nucleotides. To establish a vaccinia virus-free recovery system, a BHK-derived cell line stably expressing T7 RNA polymerase was generated (BSR T7/5). Recombinant BRSV was reproducibly recovered from cDNA constructs after T7 RNA polymerase-driven expression of antigenome sense RNA and of BRSV N, P, M2, and L proteins from transfected plasmids. Chimeric viruses in which the BRSV leader region was replaced by the human respiratory syncytial virus (HRSV) leader region replicated in cell culture as efficiently as their nonchimeric counterparts, demonstrating that all cis-acting sequences of the HRSV promoter are faithfully recognized by the BRSV polymerase complex. In addition, we report the successful recovery of a BRSV mutant lacking the complete NS2 gene, which encodes a nonstructural protein of unknown function. The NS2-deficient BRSV replicated autonomously and could be passaged, demonstrating that NS2 is not essential for virus replication in cell culture. However, growth of the mutant was considerably slower than and final infectious titers were reduced by a factor of at least 10 compared to wild-type BRSV, indicating that NS2 provides a supporting factor required for full replication capacity.

Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter.

Here are two books on a topic new to Technometrics: statistical and mathematical demography. The first author of Applied Mathematical Demography wrote the first two editions of this book alone. The second edition was published in 1985. Professor Keyfritz noted in the Preface (p. vii) that at age 90 he had no interest in doing another edition; however, the publisher encouraged him to find a coauthor. The result is an additional focus for the book in the world of biology that makes it much more relevant for the sciences. The book is now part of the publisher’s series on Statistics for Biology and Health. Much of it, of course, focuses on the many aspects of human populations. The new material focuses on mature population models, the particular focus of the new author (see, e.g., Caswell 2000). As one might expect from a book that was originally written in the 1970s, it does not include a lot of information on statistical computing. The new book by Alho and Spencer is focused on putting a better emphasis on statistics in the discipline of demography (Preface, p. vii). It is part of the publisher’s Series in Statistics. The authors are both statisticians, so the focus is on statistics as used for demographic problems. The authors are targeting human applications, so their perspective on science does not extend any further than epidemiology. The book actually strikes a good balance between statistical tools and demographic applications. The authors use the first two chapters to teach statisticians about the concepts of demography. The next four chapters are very similar to the statistics content found in introductory books on survival analysis, such as the recent book by Kleinbaum and Klein (2005), reported by Ziegel (2006). The next three chapters are focused on various aspects of forecasting demographic rates. The book concludes with chapters focusing on three areas of applications: errors in census numbers, financial applications, and small-area estimates.

Univariate Discrete Distributions

Wildlife diseases are in fashion. This is creating an explosion of related knowledge. Despite this, the dynamics of both wildlife and diseases and the changes in livestock and wildlife management make it increasingly difficult to overview the current situation of wildlife diseases in Europe. This paper aims to discuss the available management possibilities and to highlight current research priorities. One area that causes severe concern to authorities is diseases largely under control in domestic populations but still existing as a reservoir in wildlife. Multihost situations are also of concern for wildlife management and conservation, as diseases can affect the productivity and density of wildlife populations with an economic or recreational value. Concern about emerging diseases is rising in recent years, and these may well occur at the fertile livestock-wildlife interface. Wildlife-related zoonoses are a diverse and complex issue that requires a close collabora- tion between wildlife ecologists, veterinarians and public health professionals. A few risk factors can be identified in most of the relevant wildlife diseases. Among them are (1) the introduction of diseases through movements or trans- locations of wild or domestic animals, (2) the consequences of wildlife overabundance, (3) the risks of open air livestock breeding, (4) vector expansion and (5) the expansion or introduction of hosts. Wildlife disease control requires the integration of veterinary, ecology and wildlife management expertise. In addition to surveillance, attempts to control wildlife diseases or to avoid disease transmission between wildlife and livestock have been based on setting up barriers, culling, hygienic measures, habitat manage- ment, vector control, treatments and vaccination. Surveil- lance and descriptive studies are still valuable in regions, species or diseases that have received less attention or are (at least apparently) emerging. Nonetheless, limiting the research effort to the mere reporting of wildlife disease outbreaks is of limited value if management recommenda- tions are not given at the same time. Thus, more experimental approaches are needed to produce substantial knowledge that enables authorities to make targeted management recommendations. This requires policy mak- ers to be more aware of the value of science and to provide extra-funding for the establishment of multidisciplinary scientific teams.

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Diseases shared between wildlife and livestock: a European perspective

Bats are hosts to a variety of viruses capable of zoonotic transmissions. Because of increased contact between bats, humans, and other animal species, the possibility exists for further cross-species transmissions and ensuing disease outbreaks. We describe here full and partial viral genomes identified using metagenomics in the guano of bats from California and Texas. A total of 34% and 58% of 390,000 sequence reads from bat guano in California and Texas, respectively, were related to eukaryotic viruses, and the largest proportion of those infect insects, reflecting the diet of these insectivorous bats, including members of the viral families Dicistroviridae, Iflaviridae, Tetraviridae, and Nodaviridae and the subfamily Densovirinae. The second largest proportion of virus-related sequences infects plants and fungi, likely reflecting the diet of ingested insects, including members of the viral families Luteoviridae, Secoviridae, Tymoviridae, and Partitiviridae and the genus Sobemovirus. Bat guano viruses related to those infecting mammals comprised the third largest group, including members of the viral families Parvoviridae, Circoviridae, Picornaviridae, Adenoviridae, Poxviridae, Astroviridae, and Coronaviridae. No close relative of known human viral pathogens was identified in these bat populations. Phylogenetic analysis was used to clarify the relationship to known viral taxa of novel sequences detected in bat guano samples, showing that some guano viral sequences fall outside existing taxonomic groups. This initial characterization of the bat guano virome, the first metagenomic analysis of viruses in wild mammals using second-generation sequencing, therefore showed the presence of previously unidentified viral species, genera, and possibly families. Viral metagenomics is a useful tool for genetically characterizing viruses present in animals with the known capability of direct or indirect viral zoonosis to humans.

https://www.bu.edu/cecb/files/2009/08/Li-et-al_-bat-metagenomics1.pdf

Bat Guano Virome: Predominance of Dietary Viruses from Insects and Plants plus Novel Mammalian Viruses

Amongst the 60 viral species reported to be associated with bats, 59 are RNA viruses, which are potentially important in the generation of emerging and re-emerging infections in humans. The prime examples of these are the lyssaviruses and Henipavirus. The transmission of Nipah, Hendra and perhaps SARS coronavirus and Ebola virus to humans may involve intermediate amplification hosts such as pigs, horses, civets and primates, respectively. Understanding of the natural reservoir or introductory host, the amplifying host, the epidemic centre and at-risk human populations are crucial in the control of emerging zoonosis. The association between the bat coronaviruses and certain lyssaviruses with particular bat species implies co-evolution between specific viruses and bat hosts. Cross-infection between the huge number of bat species may generate new viruses which are able to jump the trans-mammalian species barrier more efficiently. The currently known viruses that have been found in bats are reviewed and the risks of transmission to humans are highlighted. Certain families of bats including the Pteropodidae, Molossidae, Phyllostomidae, and Vespertilionidae are most frequently associated with known human pathogens. A systematic survey of bats is warranted to better understand the ecology of these viruses.

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Bats as a continuing source of emerging infections in humans

Respiratory syncytial virus infections in human beings and in cattle

By using a monoclonal antibody directed against an epitope located on glycoprotein B of bovine herpesvirus 1 (BHV1), a simple, convenient blocking enzyme-linked immunosorbent assay (ELISA) which combines a high sensitivity with a low false-positive rate has been developed. The test can be performed at low variance on undiluted bovine serum samples. The epitope on glycoprotein B appears to be conserved, because it could be detected by immunostaining in all of 160 BHV1 isolates originating from 10 countries. In testing 215 anti-BHV1 antibody-negative and 179 anti-BHV1 antibody-positive serum samples, specificity and sensitivity were 0.96 and 0.99, respectively. This blocking ELISA is superior to a commercially available indirect ELISA and to the 24-h virus neutralization test in detecting low antibody levels in serum. In addition, this blocking ELISA is able to detect specific antibodies in serum as early as 7 days postinfection. To minimize any risk of introducing latent BHV1 carriers among noninfected cattle, this blocking ELISA would be, in our opinion, the test of choice.

A simple, specific, and highly sensitive blocking enzyme-linked immunosorbent assay for detection of antibodies to bovine herpesvirus 1.

To study the epidemiology of respiratory syncytial virus (RSV) infections during the year, the incidences of primary infections and reinfections were monitored by titrating antibodies to bovine RSV (BRSV) in cattle above 2 months of age in 6 dairy herds in the Netherlands. From August 1990 until September 1991, 884 cattle were sampled at one-month intervals. A total of 155 cattle, most under two years of age, had a primary antibody response. Antibody rises were found in 259 cattle of all ages. The highest incidences of BRSV infections were found in one period either in autumn or winter. In other seasons, primary infections were rare, whereas reinfections were not uncommon. In 5 out of the 6 herds, two seronegative sentinel calves were introduced at the end of the winter and none developed specific antibodies before the next winter. The observations strongly suggest that, in spite of regular reinfections, BRSV circulates during spring or summer at a very low level or not at all. Persistent BRSV infection in a number of cows might be a means for the virus to survive during summer, but a steady rate of reinfection of seropositive cows throughout the year at a low level might also maintain a reservoir of infectious virus. This study adds to the knowledge of frequency and timings of primary infections and reinfections of BRSV and it might contribute to the study of these issues of human RSV.

Dynamics of bovine respiratory syncytial virus infections: a longitudinal epidemiological study in dairy herds

To study European bat lyssavirus (EBLV) in bat reservoirs in the Netherlands, native bats have been tested for rabies since 1984. For all collected bats, data including species, age, sex, and date and location found were recorded. A total of 1,219 serotine bats, Eptesicus serotinus, were tested, and 251 (21%) were positive for lyssavirus antigen. Five (4%) of 129 specimens from the pond bat, Myotis dasycneme, were positive. Recently detected EBLV RNA segments encoding the nucleoprotein were sequenced and analyzed phylogenetically (45 specimens). All recent serotine bat specimens clustered with genotype 5 (EBLV1) sequences, and homologies within subgenotypes EBLV1a and EBLV1b were 99.0%-100% and 99.2%-100%, respectively. Our findings indicate that EBLVs of genotype 5 are endemic in the serotine bat in the Netherlands. Since EBLVs can cause fatal infections in humans, all serotine and pond bats involved in contact incidents should be tested to determine whether the victim was exposed to EBLVs.

/pdf/european-bat-lyssaviruses-the-netherlands-4ifdk9bnzm.pdf

European bat lyssaviruses, The Netherlands.

Bats classified in the order Chiroptera are the most abundant and widely distributed non-human mammalian species in the world. Several bat species are reservoir hosts of zoonotic viruses and therefore can be a public health hazard. Lyssaviruses of different genotypes have emerged from bats in America (Genotype 1 rabies virus; RABV), Europe (European bat lyssavirus; EBLV), and Australia (Australian bat lyssavirus; ABLV), whereas Nipah virus is the most important recent zoonosis of bat origin in Asia. Furthermore, some insectivorous bat species may be important reservoirs of SARS coronavirus, whereas Ebola virus has been detected in some megachiropteran fruit bats. Thus far, European bat lyssavirus (EBLV) is the only zoonotic virus that has been detected in bats in Europe. New zoonotic viruses may emerge from bat reservoirs and known ones may spread to a wider geographical range. To assess future threats posed by zoonotic viruses of bats, there is a need for accurate knowledge of the factors underlying disease emergence, for an effective surveillance programme, and for a rapid response system. In Europe, primary efforts should be focussed on the implementation of effective passive and active surveillance systems for EBLVs in the Serotine bat, Eptesicus serotinus, and Myotis species (i.e., M. daubentonii and M. dasycneme). Apart from that, detection methods for zoonotic viruses that may emerge from bats should be implemented. Analyses of data from surveillance studies can shed more light on the dynamics of bat viruses, (i.e., population persistence of viruses in bats). Subsequently, studies will have to be performed to assess the public health hazards of such viruses (i.e., infectivity and risk of infection to people). With the knowledge generated from this kind of research, a rapid response system can be set up to enhance public health awareness of emerging zoonotic viruses of bats.

J. A. Kramps

Papers

Respiratory syncytial virus infections in human beings and in cattle

A simple, specific, and highly sensitive blocking enzyme-linked immunosorbent assay for detection of antibodies to bovine herpesvirus 1.

Dynamics of bovine respiratory syncytial virus infections: a longitudinal epidemiological study in dairy herds

European bat lyssaviruses, The Netherlands.

Public health awareness of emerging zoonotic viruses of bats: a European perspective.