Potential zoonotic pathogens hosted by endangered bonobos.

Few publications, often limited to one specific pathogen, have studied bonobos (Pan paniscus), our closest living relatives, as possible reservoirs of certain human infectious agents. Here, 91 stool samples from semicaptive bonobos and bonobos reintroduced in the wild, in the Democratic Republic of the Congo, were screened for different infectious agents: viruses, bacteria and parasites. We showed the presence of potentially zoonotic viral, bacterial or parasitic agents in stool samples, sometimes coinfecting the same individuals. A high prevalence of Human mastadenoviruses (HAdV-C, HAdV-B, HAdV-E) was observed. Encephalomyocarditis viruses were identified in semicaptive bonobos, although identified genotypes were different from those identified in the previous fatal myocarditis epidemic at the same site in 2009. Non-pallidum Treponema spp. including symbiotic T. succinifaciens, T. berlinense and several potential new species with unknown pathogenicity were identified. We detected DNA of non-tuberculosis Mycobacterium spp., Acinetobacter spp., Salmonella spp. as well as pathogenic Leptospira interrogans. Zoonotic parasites such as Taenia solium and Strongyloides stercoralis were predominantly present in wild bonobos, while Giardia lamblia was found only in bonobos in contact with humans, suggesting a possible exchange. One third of bonobos carried Oesophagostomum spp., particularly zoonotic O. stephanostomum and O. bifurcum-like species, as well as other uncharacterized Nematoda. Trypanosoma theileri has been identified in semicaptive bonobos. Pathogens typically known to be transmitted sexually were not identified. We present here the results of a reasonably-sized screening study detecting DNA/RNA sequence evidence of potentially pathogenic viruses and microorganisms in bonobo based on a noninvasive sampling method (feces) and focused PCR diagnostics.

Prevalence and characteristics of a novel bovine adenovirus type 3 with a natural deletion fiber gene.

Bovine adenovirus type 3 (BAdV-3) is an important pathogen causing bovine respiratory disease complex (BRDC). From Jun 2016 to Jun 2018, 108 nose swab samples were collected from cattle with BRDC from 11 farms in five cities, and 78.7% (85/108) samples were detected as BAdV-3 positive by Real-time PCR. Interestingly, the sequences of 7/10 fiber (852 bp) and hexon (785 bp) fragments cloned from 10 positive samples from eight farms were clustered into a single branch of the evolutionary tree. A BAdV-3 strain (BO/YB24/17/CH) was successfully isolated. The isolate caused pathological changes of lung, trachea and spleen in BALB/c mice. Notably, 79 amino acid deletions in the shaft domain and 74 unique amino acid mutations were found in the fiber gene of the isolate compared with the available complete sequences for fiber genes in the GenBank database. These characteristics indicated that the isolate may represent a novel fiber genotype of BAdV-3. A pair of specific primers covering the deletion region in the fiber gene was designed to screen the prevalence of BAdV-3 encoding the novel fiber gene. The results showed that 7 of the 10 strains possessed the novel fiber gene, and these novel fiber strains were detected from six farms in which calves were just imported from five provinces, indicating that this BAdV-3 with the natural deletion fiber gene has a wide geographical distribution in China. In conclusion, our results reveal that BAdV-3 is widespread in China and a pathogenic BAdV-3 strain with a novel fiber gene has been detected at high frequency, which is beneficial to understand the prevalence and genetic evolution of BAdV-3.

Evaluation of the Anti-Adenoviral Activity of ALTANT, an Ozonated Alcohol Disinfectant.

Seven human mastadenovirus (HAdV) species (A-G) are known with more than 100 reported types. HAdV is highly resistant to common hand sanitizers. Epidemic keratoconjunctivitis and pharyngoconjunctival fever are caused by HAdV, which can be explosively transmitted in a confined space, resulting in outbreaks, such as nosocomial infections. Given the absence of an antiviral agent against the HAdV infection, it is important to prevent the spread of the infection by using disinfectants. Ozone has already been well-known for its bactericidal and virucidal effects. ALTANT is an ozonated alcohol preparation developed by E-TECH Co., Ltd. (Kobe, Hyogo, Japan). In this study, we mixed ALTANT with different HAdV types at a ratio of 9:1 and determined HAdV viability after instantaneous reactions for varying periods (flash to 5 minutes) using the TCID50 assay. The assay results demonstrated that the HAdV viability decreased by 1/10 to 1/100 within 1 minute after the reaction; additionally, slight differences in the reactivity were observed among the HAdV types. HAdV viability decreased by a factor of > 4log10, and the virus was eliminated within 3 minutes. This study demonstrated the potent HAdV disinfection effect of ALTANT.

Murine adenoviruses: tools for studying adenovirus pathogenesis in a natural host.

Small laboratory animals are powerful models for investigating in vivo viral pathogenesis of a number of viruses. For adenoviruses (AdVs), however, species-specificity poses limitations to studying human adenoviruses (HAdVs) in mice and other small laboratory animals. Thus, this review covers work on naturally occurring mouse AdVs, primarily mouse adenovirus type 1 (MAdV-1), a member of the species Murine mastadenovirus A. Molecular genetics, virus life cycle, cell and tissue tropism, interactions with the host immune response, persistence, and host genetics of susceptibility are described. A brief discussion of MAdV-2 (member of species Murine mastadenovirus B) and MAdV-3 (member of species Murine mastadenovirus C) is included. We report the use of MAdVs in the development of vectors and vaccines.

Molecular evolution of human adenovirus type 16 through multiple recombination events.

Human mastadenoviruses (HAdVs) are non-enveloped, double-stranded DNA viruses that are comprised of more than 85 types classified within seven species (A-G) based on genomics. All HAdV prototypes and many newly defined type genomes have been completely sequenced and are available. Computational analyses of the prototypes and newly emergent HAdV strains provide insights into the evolutionary history and molecular adaptation of HAdV. Most types of HAdV-B are important pathogens causing severe respiratory infections or urinary tract infections and are well characterized. However, HAdV-16 of the B1 subspecies has rarely been reported and its genome is poorly characterized. In this study, bioinformatics analysis, based on genome sequences obtained in GenBank, suggested that HAdV-16, a prototype HAdV-B species, evolved from multiple intertypic recombination events. HAdV-16 genome contains the hexon loop 1 to loop 2 region from HAdV-E4, the partial hexon conserved region 4 (C4) from the subspecies HAdV-B2, genome region 30,897-33,384 containing the fiber gene from SAdV-35, and other genomic parts from the subspecies HAdV-B1. Moreover, analysis of sequence similarity with HAdV-E4 LI, LII, and SAdV-36 strains demonstrated the recombination events happened rather early. Further, amino acid sequence alignment indicated that the amino acid variations occurred in hypervariable regions (HVRs). Especially, the major difference in HVR7, which contains the critical neutralization epitope of HAdV-E4, between HAdV-16 and HAdV-E4 might explain the low level of cross-neutralization between these strains. Our findings promote better understanding on HAdV evolution, predicting newly emergent HAdV strains, and developing novel HAdV vectors.

Temporal dynamics of human mastadenovirus species in cases of respiratory illness in southern Brazil

Human mastadenovirus (HAdV) genus is related to several diseases, among them upper and lower respiratory tract illness. HAdV species B, C, D, and E are mainly associated with respiratory infections. The goal of this work was to identify the HAdV species associated with respiratory infections in hospitalized patients from southern Brazil. Samples were collected from 1996 to 2004 and 2011 to 2017. During this period, 28,524 samples were collected, and 9983 were positive for respiratory viruses, being 435 for HAdV. From these 435 samples, 57 were selected for characterization of HAdV species. For screening the presence of HAdV, a partial sequence of the DNA polymerase gene (DNApol gene) was amplified by nested PCR. Partial nucleotide sequencing was performed in positive samples, and HAdV (DNApol gene) was detected in 53 samples: species B (28;49.1%), C (16;8.0%), D (2; 3.5%), E (5; 8.7%), and untyped (2; 3.5%). Specie D was found only in 2017 and specie E in 2011 and 2012. The age of the patients ranged from < 1 to 81 years old, and 62.3%were male. No relationship between gender orage and identified HAdV species were observed. In addition, in the period of 2013­2017, 18 samples from patients who died were analyzed: 11 were related to species B, 4 to C, and 2 to D and 1 remained untyped. Circulation of HAdV species D and Evaried over the years, but species B and C were present throughout the evaluated period. In addition, respiratory infections by HAdVaffect elderly and children mainly. (AU)

Development of polymerase chain reaction-based diagnostic tests for detection of Malsoor virus & adenovirus isolated from Rousettus species of bats in Maharashtra, India.

BACKGROUND & OBJECTIVES: Bats are recognized as important reservoirs for emerging infectious disease and some unknown viral diseases. Two novel viruses, Malsoor virus (family Bunyaviridae, genus, Phlebovirus) and a novel adenovirus (AdV) (family, Adenoviridae genus, Mastadenovirus), were identified from Rousettus bats in the Maharashtra State of India. This study was done to develop and optimize real time reverse transcription - polymerase chain reaction (RT-PCR) assays for Malsoor virus and real time and nested PCR for adenovirus from Rousettus bats. METHODS: For rapid and accurate screening of Malsoor virus and adenovirus a nested polymerase chain reaction and TaqMan-based real-time PCR were developed. Highly conserved region of nucleoprotein gene of phleboviruses and polymerase gene sequence from the Indian bat AdV isolate polyprotein gene were selected respectively for diagnostic assay development of Malsoor virus and AdV. Sensitivity and specificity of assays were calculated and optimized assays were used to screen bat samples. RESULTS: Molecular diagnostic assays were developed for screening of Malsoor virus and AdV and those were found to be specific. Based on the experiments performed with different parameters, nested PCR was found to be more sensitive than real-time PCR; however, for rapid screening, real-time PCR can be used and further nested PCR can be used for final confirmation or in those laboratories where real-time facility/expertise is not existing. INTERPRETATION & CONCLUSIONS: This study reports the development and optimization of nested RT-PCR and a TaqMan-based real-time PCR for Malsoor virus and AdV. The diagnostic assays can be used for rapid detection of these novel viruses to understand their prevalence among bat population.

Matrix Metalloproteinase Activity in Infections by an Encephalitic Virus, Mouse Adenovirus Type 1.

Mouse adenovirus type 1 (MAV-1) infection causes encephalitis in susceptible strains of mice and alters the permeability of infected brains to small molecules, which indicates disruption of the blood-brain barrier (BBB). Under pathological conditions, matrix metalloproteinases (MMPs) can disrupt the BBB through their proteolytic activity on basement membrane and tight junction proteins. We examined whether MAV-1 infection alters MMP activity in vivo and in vitro Infected MAV-1-susceptible SJL mice had higher MMP2 and MMP9 activity in brains, measured by gelatin zymography, than mock-infected mice. Infected MAV-1-resistant BALB/c mice had MMP activity levels equivalent to those in mock infection. Primary SJL mouse brain endothelial cells (a target of MAV-1 in vivo) infected ex vivo with MAV-1 had no difference in activities of secreted MMP2 and MMP9 from mock cells. We show for the first time that astrocytes and microglia are also infected in vivo by MAV-1. Infected mixed primary cultures of astrocytes and microglia had higher levels of MMP2 and MMP9 activity than mock-infected cells. These results indicate that increased MMP activity in the brains of MAV-1-infected susceptible mice may be due to MMP activity produced by endothelial cells, astrocytes, and microglia, which in turn may contribute to BBB disruption and encephalitis in susceptible mice.IMPORTANCE RNA and DNA viruses can cause encephalitis; in some cases, this is accompanied by MMP-mediated disruption of the BBB. Activated MMPs degrade extracellular matrix and cleave tight-junction proteins and cytokines, modulating their functions. MAV-1 infection of susceptible mice is a tractable small-animal model for encephalitis, and the virus causes disruption of the BBB. We showed that MAV-1 infection increases enzymatic activity of two key MMPs known to be secreted and activated in neuroinflammation, MMP2 and MMP9, in brains of susceptible mice. MAV-1 infects endothelial cells, astrocytes, and microglia, cell types in the neurovascular unit that can secrete MMPs. Ex vivo MAV-1 infection of these cell types caused higher MMP activity than mock infection, suggesting that they may contribute to the higher MMP activity seen in vivo To our knowledge, this provides the first evidence of an encephalitic DNA virus in its natural host causing increased MMP activity in brains.

First isolation of a new type of human adenovirus (genotype 79), species Human mastadenovirus B (B2) from sewage water in Japan.

Human mastadenoviruses (HAdVs) are highly infectious viral pathogens that survive for prolonged periods in environmental waters. We monitored the presence of HAdVs in sewage waters between April 2014 and March 2015. A total of 27 adenoviral strains were detected in 75% (18/24 in occasion-base) of 24 wastewater collected samples. We identified the types of the strains as HAdV-C2 (n = 5), HAdV-A31 (5), HAdV-C1 (4), HAdV-B3 (4), HAdV-C5 (4), HAdV-B11 (2), P11H34F11 (2), and HAdV-D56 (1). The complete genome sequence of one P11H34F11 (strain T150125) was determined by next-generation sequencing and compared to other genome sequences of HAdV-B strains. The comparisons revealed evidence of a recombination event with breaking point in the hexon encoding region, which evidenced high similarity to HAdV-B34, while half of the rest of the genome showed similarity to HAdV-B11, including regions encoding fiber and E3 region proteins. The penton base encoding region seemed to be a recombinant product of HAdV-B14, -34; however, it was evidenced to be divergent to both as a novel type despite showing low bootstrap to support a new clade. We propose T150125 (P11H34F11) is a strain of a novel genotype, HAdV-79. These results support the usefulness of environmental surveillance approaches to monitor circulating HAdVs including novel types.

Deletion of pV affects integrity of capsid causing defect in the infectivity of bovine adenovirus-3.

Members of the genus Mastadenovirus including bovine adenovirus 3 (BAdV-3) encode a genus-specific unique protein named pV. The pV encoded by BAdV-3 is a protein of 423 aa showing 40.9 % identity to pV of human adenovirus 2. Here, we report the construction and analysis of recombinant BAdV-3 (BAV.dV) containing deletion of pV. The BAV.dV could only be isolated in CRL.pV cells expressing pV, suggesting that pV appears essential for the infection of BAdV-3. Analysis of BAV.dV suggested that despite affecting some late gene expression in virus-infected cells, there was no significant difference in the incorporation of viral proteins in the mature virions. Moreover, analysis of mature virions revealed degraded capsids leading to change in morphology and infectivity of BAV.dV. Furthermore, analysis of the genome sequence of different clones of BAV.dV passaged in different cell lines revealed no mutations in core proteins pVII and pX\Mu suggesting that the replication defect may not be rescued. Our results suggest that pV is required for proper viral assembly of BAdV-3 as lack of pV produces aberrant capsids. Moreover, altered capsids lead to the production of non-infectious BAV.dV virions.

293T cells expressing simian virus 40 T antigen are semi-permissive to bovine adenovirus type 3 infection.

Human cells do not normally support productive bovine adenovirus type 3 (BAdV-3) infection. Here, the outcome of BAdV-3 infection of both 293 cells and 293 cells modified to constitutively express the simian virus 40 (SV-40) T antigen (293T cells) was studied. Whereas BAdV-3 could efficiently infect 293 cells, there was a block in virus DNA replication, late-gene expression and virus production. In contrast, replication and efficient virus production could be detected in 293T cells infected with BAdV-3 or transfected with a replication-competent genomic BAdV-3 clone (pFBAV304). Early-phase gene expression was detected readily in both BAdV-3-infected 293 and 293T cells. However, the progression to efficient viral DNA synthesis and late-phase protein synthesis occurred only in 293T cells. Electron microscopy and virus growth kinetics demonstrated the formation of progeny virus in 293T cells. The SV-40 T antigens act to overcome a barrier in BAdV-3 DNA replication in 293 cells.

Acute respiratory infection with mouse adenovirus type 1.

Studies of the pathogenesis of adenovirus respiratory disease are limited by the strict species-specificity of the adenoviruses. Following intranasal inoculation of adult C57BL/6 mice with mouse adenovirus type 1 (MAV-1), we detected MAV-1 early region 3 (E3) and hexon gene expression in the lungs at 7 days post-infection (dpi). We detected MAV-1 E3 protein in the respiratory epithelium at 7 dpi. We did not detect viral mRNA or protein at 14 dpi, but MAV-1 DNA was detected by PCR at 21 dpi. Chemokine transcript levels increased between 7 and 14 dpi in the lungs of infected mice. MAV-1 infection induced a patchy cellular infiltrate in lungs at 7 and 14 dpi. This is the first report demonstrating the presence of MAV-1 in the respiratory epithelium of infected mice and describing chemokine responses in the lung induced by MAV-1 respiratory infection. MAV-1 infection of mice has the potential to serve as a model for inflammatory changes seen in human adenovirus respiratory disease.

Role of bovine adenovirus-3 33K protein in viral replication.

The L6 region of bovine adenovirus type (BAdV)-3 encodes a nonstructural protein named 33K. To identify and characterize the 33K protein, rabbit polyclonal antiserum was raised against a 33K-GST fusion protein expressed in bacteria. Anti-33K serum immunoprecipitated a protein of 42 kDa in in vitro translated and transcribed mRNA of 33K. However, three proteins of 42, 38, and 33 kDa were detected in BAdV-3 infected cells. To determine the role of this protein in virus replication, a recombinant BAV-33S1 containing insertional inactivation of 33K (a stop codon created at the seventh amino acid of 33K ORF) was constructed. Although BAV-33S1 could be isolated, the mutant showed a severe defect in the production of progeny virus. Inactivation of the 33K gene showed no effect on early and late viral gene expression in cells infected with BAV-33S1. However, formation of mature virions was significantly reduced in cells infected with BAV-33S1. Surprisingly, insertional inactivation of 33K at amino acid 97 (pFBAV-33.KS2) proved lethal for virus production. Although expression of early or late genes was not affected, no capsid formation could be observed in mutant DNA-transfected cells. These results suggest that 33K is required for capsid assembly and efficient DNA capsid interaction.

Bovine adenovirus type 3 containing heterologous protein in the C-terminus of minor capsid protein IX.

Earlier, we detected pIX of BAdV-3 as a 14-kDa protein in purified virions. Analysis of BAdV-3 pIX using different region antibodies revealed that the N-terminus and central domain of the pIX contain immunogenic sites and are not exposed on the surface of BAdV-3 virion. This suggested that the C-terminus of BAdV-3 pIX (125 amino acid) may be exposed on the virion and may be used as a site for incorporation of heterologous peptides or proteins. We constructed recombinant BAV950 containing a small peptide (21 amino acid), including the RGD motif or recombinant BAV951 containing enhanced yellow-green fluorescent protein (EYFP) fused to the C-terminus of pIX. Western blot analysis demonstrated that the chimeric pIX-RGD was incorporated into virion capsids. Incorporation of the RGD motif into the pIX resulted in significant augmentation of BAdV-3 fiber knob-independent infection of the integrin-positive cells, suggesting that RGD motifs are displayed on the surface of virion capsids and are accessible for binding to integrins. Analysis of BAV951 revealed that the chimeric pIX is incorporated into virion capsids and EYFP containing the C-terminus of pIX is exposed on the surface of the virion. Moreover, insertion of chimeric pIXs was maintained without change through successive rounds of viral replication. These results suggested that in contrast to major capsid proteins (hexon, penton, fiber), the minor capsid protein IX can be use for the incorporation of targeting ligands based on either small peptides or longer polypeptides.

Apoptosis in calf pneumonia induced by endobronchial inoculation with bovine adenovirus type 3 (BAV-3).

Three calves aged 1 week (group 1), three aged 6 weeks (group 2) and three aged 6 weeks (having been pretreated with dexamethasone) (group 3) were infected endobronchially with bovine adenovirus type 3 (BAV-3). All calves had received colostrum. The histopathological, immunohistochemical, ultrastructural and TUNEL features were examined on post-inoculation day (PID) 3, 5 and 7. Viral replication and intranuclear inclusions were frequently observed in groups 1 and 3, but not in group 2. The lesions became progressively severe on PID 5 and 7 in group 1. In group 3, however, the cellular injury caused by BAV-3 was of short duration and the lesions began to resolve at PID 7. Numerous apoptotic cells were seen in the PID 3 calves of all three groups, and in the PID 7 calves of groups 2 and 3; however, the PID 5 and 7 calves of group 1 showed only a few apoptotic cells in the alveolar septa. The results indicated that (1) the durability of BAV-3 infection in the lung was closely related to apoptosis, and (2) the host defence mechanism that induced apoptosis in infected cells was age-related.

Effect of adenovirus type 1 and influenza A virus on Streptococcus pneumoniae nasopharyngeal colonization and otitis media in the chinchilla.

Considerable evidence has implicated respiratory tract virus potentiation of bacterial adherence, colonization, and superinfection as a significant factor contributing to the pathogenesis of otitis media (OM). Influenza A and B viruses, adenovirus, and respiratory syncytial virus are the primary respiratory tract viruses associated with this disease. Investigations have established a dramatic increase in the development of experimental OM in chinchillas co-inoculated with influenza A virus and Streptococcus pneumoniae (Spn). The mechanism underlying this phenomenon was suggested to involve, in part, viral compromise of eustachian tube mucosal integrity and function. This study was designed to assess and compare the effect of adenovirus and influenza A virus infection on adherence, the kinetics of colonization, and invasion of the middle ear by Spn in the chinchilla model of OM. Cohorts were inoculated intranasally with adenovirus type 1 or influenza A virus, and then inoculated intranasally 7 days later with Spn 6A. All cohorts were observed over a 14-day period after challenge with Spn, and the incidence and severity of OM were assessed by several methods, including culture of the nasopharynx and middle ear effusions. The data indicated that influenza A virus promotes a significant increase in nasopharyngeal colonization by Spn, an increased incidence and severity of OM, and a sustained presence of Spn in the effusions. Adenovirus infection, however, did not enhance colonization by Spn or result in an increased incidence or severity of OM.

The role of the L4 33K gene in adenovirus infection.

The late phase of adenovirus infection is characterized not only by the synthesis of late proteins and the assembly of new virions, but also by the inhibition of early gene expression and host cell translation. Previous work has demonstrated that both of these inhibitory effects depend upon expression from the major late transcription unit (MLTU), controlled by the major late promoter (MLP). Furthermore, the repression of early gene expression has been shown to be mediated in trans, suggesting a role for one or more MLTU-encoded soluble factor(s). A possible candidate for such a factor is the L4-encoded 33K gene product, a protein conserved throughout the Mastadenoviridae, but of no known function. To test the role of this protein in viral infection, a stop codon was placed at the 20th position of the 33K ORF. Viable virus with genomes containing the mutation were recovered in an overlap recombination assay. Phenotypic analysis revealed that the mutant virus had a significant deficiency in both kinetics of replication and final yield, as compared to the wild-type virus. Detailed analysis of infected cells showed that there was no detectable change in the regulation of expression of several early genes and the pIX gene. This suggests either that 33K is not involved in this late phase phenomenon or that this function is replaceable by another late protein(s). Late protein synthesis and accumulation were similar to those in wild-type-infected cells. However, the reduced yield of infectious mutant virus could be accounted for by a marked deficiency in the accumulation of intermediate particles and completed capsids, suggesting a role for 33K in the process of assembly. In addition there was a small but reproducible deficiency in the shutoff of host cell translation. These results show that the 33K protein plays an important, although apparently not essential, function in the late phase of virus infection.

[The patterns in the molecular biology of the adenoviruses (the results of a 20-year study at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine)]. / Nekotorye zakonomernosti molekuliarnoi biologii adenovirusov (itogi 20-letnego izlucheniia v Institute mikrobiologii i virusologii NAN Ukrainy).

Scientific data of the highest importance and priority concerning regularities of structural and functional organization of proteins of adenoviruses capsids and peculiarities of expression of the virus genome are as follows: New antigen determinants of hexon and adenovirus fiber have been discovered, their different nature (conformational or linear) and different orientation, depending on the spatial organization of proteins, have been proved; localization of some epitopes has been determined with the help of synthesised antigen-active peptides, imitating them. Some regularities of structural and functional organization of adenovirus hexon have been determined on the basis of comparative analysis of antigenic specificity and primary structure of proteins being apart in taxonomic respect. The conception of immunoactivation (infectivity neutralization) of adenoviruses has been developed, and a mathematical model of this process has been first proposed, which determines the impact of antibodies to several antigenic determinants of hexon and fiber as well as interferon and complement. The unknown peculiarities of the adenovirus genome expression were studied in the dynamics of productive infection or under the effect of modified nucleosides, proteolysis inhibitors and those of different nature promising for chemotherapy of adenovirus infection. Lymphotropicity of adenoviruses was established and a model of the mixed infection of lymphocytes with adenoviruses, HIV, and Epstein-Barr virus of the herpes virus family was proposed for the first time. It was determined that the mutual interference of viruses was developed at the process of a single or successive infection and this was important to understand AIDS immunopathogenesis. Data presented substantiate the ways of creation of modern efficient preparations for diagnosis, prophylaxis and chemotherapy of adenovirus infection.

Generation of infectious genome of bovine adenovirus type 3 by homologous recombination in bacteria.

The widely used technique of generating adenovirus vectors by homologous recombination in mammalian cells is usually not very efficient. This communication describes a simple method of generating a plasmid containing the full-length genome of an adenovirus by homologous recombination in bacteria. Following transfection of a suitable mammalian cell line with the full-length adenovirus genome, infectious virus progeny could easily be generated. Using this technique the generation of adenovirus recombinants would be efficient and straightforward.

Isolation of a bovine adenovirus serotype 10 from a calf in the United States.

Virus isolated from the lung, liver, kidney, and small intestine of a 3-month-old Holstein heifer with a clinical history of pneumonia and lesions in multiple organs was identified as an adenovirus on the basis of morphological and physicochemical characteristics. The adenovirus was determined to be a serotype 10 bovine adenovirus and represents the first reported isolation of this serotype in the United States. Inoculation of calves with this isolate resulted in mild to moderate clinical response consisting of fever, inappetence, increased respiratory rate, cough, and listlessness. Gross lesions were minimal in the respiratory tract and consisted of fibrin in the airways and small areas of consolidation in the cranial lobes of the lung. Mucofibrinous foci were present on the mucosa of the upper small intestine.