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M Benkö

Publications and source records attributed to M Benkö.

At least 19 recordsLinked to original sources

Isolation of bovine adenovirus serotype 6 from a calf in the United Kingdom.

Two viruses, designated 99-8130(C) and 99-8130(I), were isolated in calf testis cells from the colon and ileum, respectively, of a suckled beef calf which had developed dysentery and died. Electron microscopy indicated that the mean (sd) size of the viral particles, 83 (2.5) nm, and their morphology were consistent with their being members of the family Adenoviridae. They were confirmed as adenoviruses by PCR when products of the expected size (608 bp) were amplified from both isolates by using a primer pair specific for members of the genus Atadenovirus. A comparison of the sequence of a 567 bp segment of the 99-8130(C) amplicon with that of other prototype bovine adenovirus (BAdV) strains of atadenoviruses identified the isolate as BAdV serotype 6 (BAdV-6), which had 99.3 per cent and 100 per cent identities at the nucleotide and amino acid levels, respectively, with the prototype BAdV-6 strain 671130. A virus neutralisation test was developed and indicated a high prevalence of antibody to BAdV-6 in Northern Irish cattle. There was no evidence of adenoviral inclusions in tissues from the affected calf and no antigen was detected when the tissues were stained by an immunoperoxidase technique, using a homologous antiserum raised in rabbits. The two viruses were the third reported isolation of BAdV-6, and the first from a clinically ill bovine animal.

Adenoviridae↗

Molecular evolution of adenoviruses.

New advances in the field of genetic characterization of adenoviruses originating from different animal species are summarized. Variations seen in the host range and specificity, pathogenicity, genomic arrangement or gene complement are much wider than expected based on previous studies of human adenoviruses. Several exceptional adenoviruses from the two traditional conventional genera are now removed, and proposed to form at least two new genera. The eventual host origin of the new genera, however, is not clarified. Novel results from the genomic and phylogenetic analyses of adenoviruses originating from lower vertebrate species (including reptiles, amphibians and fish) seem to imply that probably five major clusters of adenoviruses exist corresponding to the five major classes of Vertebrata. Adenoviruses, which are now suspected to have common origin with enterobacterium phages from the family Tectiviridae, are perhaps very ancient indeed, and may have undergone a co-evolution with vertebrate hosts.

Adenoviridae↗

DNA sequence of a small, unidentified plasmid isolated from a Haemophilus somnus strain: short communication.

One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cattle with respiratory disease was purified and cloned for DNA sequencing. The plasmid was found to be 1065 base pairs long with 39.2% G + C content, and showed no homology to any DNA sequenced so far. It has no capacity to code any protein longer than 43 residues. It is not clear yet if this plasmid codes Haemophilus somnus specific factors.

Animals↗

DNA sequencing and phylogenetic analysis of the protease gene of ovine adenovirus 3 suggest that adenoviruses of sheep belong to two different genera.

Until now, the only published ovine adenovirus DNA sequence was the complete genome of ovine adenovirus isolate 287 (OAV287) which, compared to other mammalian adenoviruses, possesses strikingly unique genomic organisation and should properly be classified into a new adenovirus genus. The protease gene sequence of ovine adenovirus type 3 (OAdV-3) was determined and analysed. The results of phylogenetic analysis of the 205 residue long protein demonstrated that OAdV-3 belongs to the genus Mastadenovirus, and is surprisingly closely related to bovine adenovirus type 2. In spite of the common host origin, the evolutionary distance between OAdV-3 and OAV287 proved to be great suggesting that sheep, similarly to cattle and fowl, might be infected by distantly related adenoviruses belonging to different genera.

Adenoviridae Infections↗

Comparison of the genome of ovine adenovirus types 1 through 5 by restriction enzyme analysis and DNA hybridisation.

The DNA of the prototype strains of ovine adenovirus (OAdV) 1 through 5 was analysed by restriction enzyme (RE) digestion. The RE patterns generated by HindIII and PstI enzymes were characteristic of the examined strains. OAdV-2 and 3 resembled each other the most, and their EcoRI and HindIII patterns seemed to be identical. Considering the number of comigrating fragments, serotypes OAdV-2, 3, 4 and 5 looked more closely related to each other than to OAdV-1. This finding was strengthened by Southern blot hybridisations probed with random HindIII clones of OAdV-3. The estimated genome size of the examined OAdV types ranged between 31.9 and 32.8 kilobase pairs. The results supported the new genus classification of OAdVs.

Animals↗

Sequence, transcriptional analysis, and deletion of the bovine adenovirus type 1 E3 region.

The early 3 (E3) transcriptional unit of human adenoviruses (HAV) encodes proteins that modulate host antiviral immune defenses. HAV E3 sequences are highly variable; different HAV groups encode phylogenetically unrelated proteins. The role of the E3 region of many human and animal adenoviruses is unknown because the sequences are unrelated to previously characterized viruses and the functions of proteins encoded by these regions have not been studied. We sequenced a portion of the bovine adenovirus serotype 1 (BAV-1) genome corresponding to the putative E3 region. This sequence was substantially different from other adenoviral E3 sequences, including those of two other bovine adenoviruses. However, two regions of putative sequence conservation were identified. BAV-1 E3 sequences were identified in early and late transcripts, but, unlike HAV, introns were not detected in the E3 region transcripts. Like HAV E3, a majority of the BAV-1 E3 region was not essential for growth in cell culture, as demonstrated by the construction of a recombinant BAV-1 lacking 60% of the putative E3 region.

Adenovirus E3 Proteins↗

A proposal for a new (third) genus within the family Adenoviridae.

This article presents a proposal for the establishment of a new adenovirus genus to accommodate certain bovine, ovine, and avian adenoviruses with special characteristics which differentiate them from members of the existing genera Mastadenovirus and Aviadenovirus. This proposal has been developed from earlier versions with advice from the Adenovirus Study Group of the International Committee on Taxonomy of Viruses (ICTV).

Adenoviridae↗

Sequencing and phylogenetic analysis of the protease gene, and genetic mapping of bovine adenovirus type 10 define its relatedness to other bovine adenoviruses.

The complete genome of a bovine adenovirus (BAV) type 10 isolate was molecularly cloned and partially sequenced. The encoded proteins were predicted by computer analysis of the DNA sequences of the ends or the entire length of the cloned viral fragments, and thus a rough genetic map was constructed. The protease gene of BAV-10 was completely sequenced and used in phylogenetic analysis. Based on the results of the phylogenetic analysis, and the location and presence of certain genes thought to be specifically characteristic of subgroup 1 or subgroup 2 BAVs, it could be concluded that, in spite of the striking similarity in certain biological properties, BAV-10 is not related to subgroup 2 BAVs as originally described. It does not however fit clearly into subgroup 1 either, the members of which show closer relationship with human adenoviruses. BAV-10 therefore should best be considered as the first member of a third subgroup of BAVs.

Adenoviruses, Canine↗

Analysis of the hexon gene sequence of bovine adenovirus type 4 provides further support for a new adenovirus genus (Atadenovirus).

The putative hexon gene of bovine adenovirus type 4 (BAV-4), encoding 910 amino acid residues, has been identified and sequenced. A characteristic codon usage biased towards the use of AT-rich triplets was observed. Comparative analysis with other hexon sequences detected a high level of amino acid identity in the regions corresponding to the pedestals of the hexon. Substitutions, insertions and deletions were identified mainly in the variable regions forming the loops which are exposed on the outer surface of the virion. In these variable regions, BAV-4 shared similarity only with egg drop syndrome (EDS) virus and ovine adenovirus isolate 287 (OAV287). The close relationship of these viruses was also demonstrated by phylogenetic analysis of the hexon gene. In addition to the two groups of the Mastadenovirus and Aviadenovirus genera, a third cluster appeared comprising BAV-4, OAV287 and EDS virus.

Amino Acid Sequence↗

Pathological and in situ hybridisation findings in calves experimentally infected with bovine adenovirus type 4.

Three colostrum deprived calves were inoculated intravenously with a strain of bovine adenovirus type 4 (BAV-4) isolated from a calf with acute fibrinous enteritis. The calves were monitored clinically and submitted to post mortem examination at 5, 6 and 10 days post inoculation (p. i.) respectively. The 3 animals showed raise in temperature and diarrhoea. Acute enteritis was observed in calves submitted to post mortem examination at 5 and 6 days p. i. In these animals viral DNA was specifically detected on histological sections by an in situ hybridization (ISH) technique; BAV-4 was isolated from most of the organs and numerous adenovirus-positive endothelial cells were present in the gastrointestinal tract, mesenteric lymph nodes and liver. In the calf examined 10 days p. i. no lesions were observed and rare ISH positive cells were detected. These results indicate that in experimental conditions our strain of BAV-4 is capable to induce clinical signs and lesions similar to those reported in field cases.

Adenoviridae Infections↗

Close phylogenetic relationship between egg drop syndrome virus, bovine adenovirus serotype 7, and ovine adenovirus strain 287.

A cloned egg drop syndrome (EDS) virus genomic DNA fragment containing the protease gene has been identified and the complete nucleotide sequence of the protease and partial nucleotide sequence of the hexon genes has been determined. Phylogenetic analysis of the protease gene has revealed EDS virus to be genetically more closely related to bovine adenovirus type 7 (BAV-7) and ovine adenovirus isolate 287 (OAV287) than either of these two viruses are to other members of the genus Mastadenovirus or EDS virus is to an other member of the Aviadenovirus genus. The three viruses share further similarities in that they have a high percentage AT content in their genome and are characterized by having more compact genomes than other adenoviruses. The protease gene from all three viruses contained the active site residues (H55-D72-C122 triad) and C104 (providing a disulfide bond to cofactor pVIc). However, P137, found in all other members of the Mastadenovirus genus, and thought to be involved in trafficking, was missing from the protease of the EDS virus, BAV-7, and OAV287. These results suggest that EDS virus should be classified along with BAV-7 and OAV287 in a separate taxon.

Adenoviridae↗

Bovine adenovirus type 10 identified in fatal cases of adenovirus-associated enteric disease in cattle by in situ hybridization.

A severe, naturally occurring enteric disease of cattle in which adenovirus inclusions are present in the intestinal vascular endothelium has been recognized in several countries; three different adenovirus serotypes have been isolated from affected animals. An in situ hybridization technique for the detection of bovine adenoviral DNA was developed and applied to tissue from 13 cattle in Northern Ireland diagnosed to have the disease. Bovine adenovirus serotype 10 (BAV-10) was identified in the vascular inclusions of all cattle, providing strong evidence that adenoviral enteric vascular disease in cattle is associated with this serotype. The existence of BAV-10 has only recently been recognized. The first molecular biology-based technique for the diagnosis of BAV-10 infection is described. The animals in the present study are the first in which BAV-10 has had a confirmed role in a pathologic process.

Adenoviridae↗

Detection of homologous DNA sequences in animal adenoviruses by polymerase chain reaction.

A primer pair originally designed for the universal detection of human adenovirus (HAV) serotypes of all subgenera was modified then tested and found feasible for the detection of different bovine, ovine, and porcine adenovirus (BAV, OAV, and PAV, respectively) serotypes. Apparently, in the examined viruses, parts of the DNA sequence coding for the basal part of the hexon protein are conserved enough for being applicable in polymerase chain reaction (PCR) as primers. Positive amplification could be obtained even from the so-called subgroup 2 BAVs, which viruses do not cross react with HAVs or subgroup 1 BAVs in Southern hybridisation.

Adenoviruses, Human↗

Restriction site mapping of a bovine adenovirus type 10 strain.

Physical maps of the genome of a bovine adenovirus (BAV) type 10 strain (isolate Belfast1) were constructed for eight restriction enzymes. The size of the viral genome was estimated to be around 29,800 base pairs (bp). The orientation of the maps was determined on the basis of partial DNA sequences of the cloned PstI/D fragment of Belfast1. This work is an introduction to DNA sequence and phylogenetic analysis of BAV-10 in order to clarify its taxonomic place.

Adenoviridae↗

Synthesis and processing of the haemagglutinin-esterase glycoprotein of bovine coronavirus encoded in the E3 region of adenovirus.

The haemagglutinin-esterase gene (HE) of bovine coronavirus (BCV) encodes a major viral membrane glycoprotein that elicits BCV-neutralizing antibodies. The BCV HE gene was cloned into a human adenovirus serotype 5 (Ad5) transfer vector in place of early transcription region 3, and a helper-independent recombinant virus was constructed by rescue of the transcription unit by homologous in vivo recombination between the vector and Ad5 genomic DNA. The BCV HE polypeptide expressed by this recombinant Ad was characterized in vivo and in vitro. A 65K polypeptide was identified using an anti-BCV antibody in both human (293) and bovine (MDBK) cells infected with the recombinant Ad. In the absence of a reducing agent, migration of the 65K polypeptide was shifted to 130K, indicating that the recombinant HE polypeptide existed in a dimeric form. The HE polypeptide was glycosylated, as demonstrated by labelling with [3H]glucosamine, and was immunoreactive with three distinct groups of conformation-specific anti-HE monoclonal antibodies (MAbs). Cells infected with recombinant Ad expressing BCV HE exhibited both haemadsorption activity and acetylesterase activity. In addition, the anti-HE group A MAbs HC10-5 and KD9-40 inhibited both the haemadsorption activity and esterase activity of the recombinant HE polypeptide, suggesting that the antigenic domain responsible for BCV neutralization may overlap (or is closely associated with) the domain(s) responsible for haemagglutination and/or acetylesterase activities. When mice were inoculated intraperitoneally with live recombinant Ad, a significant level of BCV-neutralizing HE-specific antibody was induced. These results indicate that the recombinant Ad replicates and directs the synthesis of the BCV HE polypeptide in vivo.

Adenoviridae↗

Molecular cloning and physical mapping of the DNA of bovine adenovirus serotype 4; study of the DNA homology among bovine, human and porcine adenoviruses.

The DNA of bovine adenovirus (BAV) serotype 4, a member of the subgroup 2 BAVs, has been cloned and mapped with 11 restriction enzymes. Southern blot hybridizations probed by a clone containing about 50% of the BAV-4 genome revealed a very strong and extended DNA sequence homology amongst the members of subgroup 2, but no homology was detectable to the subgroup 1 bovine, or any of those human (HAV) and porcine adenovirus serotypes examined. These findings were strengthened by reciprocal hybridizations. When using the cloned hexon gene region of BAV-3 (subgroup 1) or the total genome of HAV-2 as probes, again no homology could be shown to the bovine subgroup 2 serotypes. The extent of DNA homology detectable between the members of bovine subgroup 1, the porcine and the human serotypes was variable, but in general less expressed than that observed within the bovine subgroup 2.

Adenoviridae↗

Restriction site mapping of bovine adenovirus type 1.

Physical maps of the DNA of bovine adenovirus (BAV) type 1 were established using ApaI, BamHI, BstEII, EcoRI and KpnI enzymes. The size of the viral genome was found to be around 35,000 base pairs. The orientation of the maps was determined by hybridizing Southern blots of BAV-1 DNA with the cloned hexon gene region of BAV-3.

Adenoviridae↗

Alternating one-letter symbols for the representation of codon variations at the level of amino acid sequences.

An idea is presented to use different letter types (e.g. L. 1, L, 1, L, l) for the alternating triplets coding the same amino acids. Such compact and demonstrative amino acid sequence comparisons can make easy the identification of differences at the DNA level if not seen in amino acid sequences (e.g. for choosing specific or non-specific primers for polymerase chain reaction).

Amino Acid Sequence↗