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Characterization of Newcastle disease virus vaccines by biological properties and sequence analysis of the hemagglutinin-neuraminidase protein gene.

Six commercially available monovalent Newcastle disease virus (NDV) live-vaccines were examined for their biological and genomic stability in comparison to their stated parent virus. Thermostability of the hemagglutinin at 56 degrees C for 5 min was consistently observed among the majority of the vaccine viruses. One exception was a recently developed NDV vaccine isolated from turkeys that had a thermostability of 15 min. Neuraminidase activity, as measured by elution rate of agglutinated red blood cells, varied among vaccine viruses and correlated with that of the parent isolate. Virulence as measured by intracerebral pathogenicity index ranged from 0 to 0.39 among NDV vaccine-type viruses, well within the range of avirulent lentogens. Sequence of the fusion protein cleavage site from all the NDV vaccine isolates examined was consistent with that for lentogens. The entire hemagglutinin-neuraminidase gene sequence was 98% similar among all the NDV vaccine viruses examined and phylogenetic classification of commercial vaccine types correlated with their respective parent virus. Consequently, the commercially produced NDV vaccines reported here appear relatively stable when mass produced in avian embryonated eggs.

Amino Acid Sequence↗

[Molecular epidemiology of Xinjiang hemorrhagic fever viruses].

BACKGROUND: To study the molecular biology of Xinjiang hemorrhagic fever (XHF) viruses, to explore its relationship with other Crimean-Congo hemorrhagic fever viruses, analyzing the epidemic origin and the tendency of geographic distribution of XHF. METHODS: The S partial segment from the patient and tick samples collected in 2001 and 2002 was tested by RT-PCR, the positive samples were sequenced directly. The nucleotide homology of S partial segment as well as the whole segments were analyzed and the phylogenetic tree of S and M gene segments was drawn by computer. RESULTS: All compared sequences of S partial segments from the patient and tick samples showed a high homology of nucleotide sequences. Phylogenetic tree divided all the analyzed viruses into three groups; Europe, African and Asian group. The Asian group can be divided further into another two branches: the middle Asian branch and the Chinese branch. All the Chinese isolates were clustered into one single group and was easy to be discriminated from the other isolates. The dividing of M segments seemed not completely related to the geographic origin of the viruses. CONCLUSION: M segment classification was not consistent to the geographic distribution of the viruses. S segments analysis showed the close relationship of genetic background between the patient isolates and the tick isolates. Besides, all the Chinese isolates have the common evolution route and the gene structure characteristics displayed the regional distribution pattern.

Animals↗

Evolutionary relationships among California serogroup viruses (Bunyaviridae) and Aedes mosquitoes (Diptera: Culicidae)

More than 100 published and four unpublished papers recording virus isolations of California (CAL) serogroup viruses from mosquitoes and other arthropods were analyzed, followed by a comparison of a contemporary classification of mosquitoes of the genus Aedes with a contemporary classification of CAL serogroup viruses. Using criteria of five or more total isolations of a given virus from a given species of mosquito, and a minimum infection rate of 1:10,000 or greater (more frequent), Aedes, and to a lesser extent Culiseta, species were found to be the arthropods most frequently involved as vectors. Such a comparison could not explain all virus-vector associations on the basis of coevolution of viruses in Aedes mosquito species, probably because of the crossing over of viruses to other mosquito species through horizontal transmission and because of deficiencies in currently available classifications. Coevolution of host-parasite systems is discussed, as are the inadequacies of contemporary taxonomic treatments of viruses and Aedes mosquitoes. The paper ends with hypothetical reconstructions of virus-mosquito relationships over time and possible routes of dispersal of California serogroup viruses to their present geographical distributions.

Aedes↗

Genetic typing of bovine viral diarrhoea virus: most Slovenian isolates are of genotypes 1d and 1f.

A selection of 43 bovine viral diarrhoea viruses isolated from mainly persistently infected cattle on 23 Slovenian farms between 1997 and 2001 were characterised genetically. Viral RNA was extracted from infected cell cultures, reverse transcribed and amplified by PCR with primers targeting the 5'-UTR and the N(pro) gene, followed by direct sequencing of purified PCR products obtained for both genomic regions. The N(pro) sequences provided the best genetic resolution, and gave also higher statistical support for phylogenetic classification of the viruses. Thirty-eight of the Slovenian isolates were of genetic subtypes 1d and 1f, four were 1b, and one subtype 1g. No BVDV type 2 viruses were found. This genetic prevalence matched those previously reported for neighbouring countries, as opposed to findings reported for more distant European countries, e.g. France, Spain and the UK. From eight cattle herds several virus isolates were analysed; with one exception all isolates from each herd were of the same genetic group. Extended sequencing of the N(pro) and part of the C gene of virus isolates with identical 5'-UTR sequences allowed differentiation between isolates obtained at different times from one herd.

5' Untranslated Regions↗

The properties of two recent isolates of cytoplasmic polyhedrosis viruses.

Cytoplasmic polyhedrosis viruses (CPVs) were isolated from laboratory cultures of Bombyx mori and Spodoptera exempta. The electrophoretic profile of the RNA segments of both viruses showed similarities with that of type 1 CPV. However, whereas the virus isolate from B. mori appeared identical to type 1 CPV (as assessed by serology, as well as the comparative mobilities of RNA segments and structural polypeptides), the isolate from S. exempta was distinct. Three of the ten viral RNA segments did not co-run with type 1 RNA, and the structural polypeptides of polyhedra had different molecular weights. It is proposed that the S. exempta isolate be included as a new 'type' (type 12) in a classification of these viruses.

Antigens, Viral↗

Electron microscopy of equine infectious anemia virus.

Equine infectious anemia (EIA) virus was observed in thin sections of infected cultured horse leukocytes by electron microscopy. The virus particles had a spherical shape and were between 80 and 120 nm in diameter. Most of them contained an electron-dense nucleoid 40 to 60 nm in diameter. They were observed to form by a process of budding from the plasma membrane and appeared to have thin surface projections. The particles described were not detected in uninfected cultured cells, and their appearance could be prevented by adding EIA immune serum to the inoculum. The implications of these findings in the classification of EIA virus are discussed.

Animals↗

Tick-borne flavivirus NS1 gene: identification of conserved peptides and antigenic analysis of recombinant louping ill virus NS1 protein.

The nucleotide sequence of the NS1 gene of louping ill (LI) virus has been determined. The sequence shows a high degree of homology with other members of the tick-borne serocomplex of flaviviruses and a lower homology with the mosquito-borne flaviviruses. Alignment of the deduced NS1 amino acid sequences with all tick-borne flavivirus NS1 sequences, identified four peptide regions which were conserved for all tick-borne flaviviruses, but were variable amongst mosquito-borne flaviviruses. A dendrogram, derived from the alignment of the NS1 protein sequences, indicated an evolutionary relationship that quite closely reflects the recognised serological classification. The LI virus NS1 protein expressed in Escherichia coli and baculoviruses showed similar antigenic reactivity to the authentic virus-coded protein when tested with NS1-specific monoclonal antibodies, but did not form high molecular weight complexes and was not secreted from cells.

Amino Acid Sequence↗

Genomic analysis of some Japanese isolates of Getah virus.

Using the reverse transcription-polymerase chain reaction (RT-PCR) and direct sequencing, capsid protein and non-structural protein 1 (nsP1) regions of Sagiyama virus and eight Getah virus strains were analysed. The viruses were isolated from Malaysia and various areas of Japan over a period of 30 years. Based on the available published sequence data, oligonucleotide primers were designed for RT-PCR and the sequences were determined. Our findings showed that though there were differences in the nucleotide sequences in the nsP1 region, there was 100% amino acid homology. On the other hand, in the capsid region, the nucleotide differences caused a major difference in the amino acid sequence. Therefore, the difference in the capsid region is one of the useful markers in the genetic classification between Sagiyama virus and strains of Getah virus, and might be responsible for the serological difference in complement fixation test. The genomic differences among the Getah virus strains are due to time factor rather than geographical distribution.

Alphavirus↗

An epidemic of paralytic poliomyelitis characterized by dual infections with poliomyelitis and Coxsackie viruses.

The first known epidemic of poliomyelitis in Easton, Pennsylvania, occurred in 1949, and was unusual in the high proportion of paralytic to non-paralytic cases. Both poliomyelitis and Cosxackie, or C, viruses were isolated from more than half the patients studied during the acute stage of the disease. One month later C virus was only occasionally recovered. Classification of the 28 strains of C virus which were isolated revealed that 24 belonged to one antigenic type, Easton-2 (related to Albany type 1 virus). Patients from whom C virus was isolated showed a rise during convalescence to the Easton-2 or homologous type antibody. Two patients with paralytic poliomyelitis were studied for the quantitative development of antibodies to the poliomyelitis virus and to the C virus found in their stools. Using the neutralization test in monkeys and in newborn mice, respectively, a simultaneous rise in antibodies to both agents was observed. The situation at present can be summarized as follows:-Poliomyelitis virus or C virus may produce infection in man, with a specific antibody response. Both agents may be carried, particularly in the intestines, without causing any serious illness and healthy carrier states have been observed for each. Both viruses can be found in nature in flies and in sewage. However there has been no evidence to suggest that these two viruses bear a relationship to each other, even when isolated from the same patient. Thus, when both viruses are found in a patient with paralysis, it is not yet possible to say with any degree of accuracy to what extent each is responsible in the over-all pattern of the disease. How frequently dual infections of this nature may occur remains for future investigations to determine. Certainly all cases of poliomyelitis are not complicated by a superimposed infection with a C virus. However, this will have to be one more item to consider in epidemic poliomyelitis.

Animals↗

Mitogen therapy for biological warfare/terrorist attacks and viral hemorrhagic fever control.

Ken Alibek was for 17 years a leader in Biopreparat, the Soviet Union's top secret agency involved in developing and stockpiling the most lethal bacteria, viruses, and toxins in the history of mankind before he defected with his family to the United States in 1992. Very contrite when he discovered he had been misled to believe that his efforts had been essential to the survival of his homeland, Alibek has become active sounding an alarm about, among other things, thousands of unemployed Russian scientists who have been seeking survival by selling their destructive expertise to rouge states and bioterrorists. Working full time in devising protective measures that might help control the damaging effects of terrorist attacks, Alibek has placed strong emphasis on stimulating nonspecific immunities of victims mainly with interleukins and other cytokines. A more productive alternative would be giving mitogens such as PHA and PWM to reinforce vaccine and antibiotic actions, at the same time stimulating protective immune, myelopoietic, and lymphopoietic responses. A key objective would be to find an effective management for the dreaded viral hemorrhagic fevers. Using Ebola infection as an experimental model, Yang et al. have shown that PHA can block both the viral secretions that inhibit neutrophil immune responses and the viral transmembrane glycoprotein that facilitates damage of the human endothelial cells responsible for the lethal hemorrhagic manifestations. Normal serum glycoproteins have in the past been clearly shown to inhibit the functions of PHA, thereby increasing dosage requirements. Extrapolation of this interaction with serum glycoproteins suggests that PHA given intravenously in adequate dosage should readily be able to block the deleterious Ebola virus glycoprotein effects. Data in an extensive classification of the hemorrhagic fever viruses recently presented by Barry make it possible to predict that mitogen therapy should be effective for virtually all of the disorders included. Therapeutic trials should best start with intravenous administration of PHA since this is the mitogen about which most is known and the only one given to humans, although the nonagglutinating advantages of fraction i.v. of PHA should be evaluated as a replacement. Functioning in a different mode, PWM has the advantage of much greater potency, and can be given either intravenously or orally, since these appear to be equally effective routes of administration. The best means of properly integrating the use of these mitogens needs to be determined.

Adjuvants, Immunologic↗

Characterization of Mycoplasmatales virus-laidlawii 3.

Purified preparations of Mycoplasmatales virus-laidlawii 3 were negatively stained and studied by electron microscopy. They were seen to consist of uniform sized particles having a polyhedral head, 57 nm by 61 nm, and a short tail, 25 nm long, joined to the head at one vertex by a collar. The particles were shown to have buoyant densities of 1-477 g/ml in CsC1, 1-32 g/ml and 1-26 g/ml in potassium tartrate and a sedimentation coefficient, S20,W, OF 290 +/- 13. They are composed of 35-2% double-stranded DNA and five structural polypeptides with approximate mol. wt. of 172000, 81000 73000, 68000 and 43000. The classification of the virus from its morphology and chemical properties is discussed.

Acholeplasma laidlawii↗

Morphogenesis of avian infectious bronchitis virus and a related human virus (strain 229E).

Avian infectious bronchitis virus (IBV) and strain 229E, a virus recently recovered from patients with colds, have been shown to possess a similar distinctive morphology in negatively stained preparations. An electron microscopic study of the morphogenesis of IBV in the chorioallantoic membrane and of strain 229E in WI-38 cells was performed. In infected cells, round electron-dense particles 82 mmu in diameter were observed to form by a process of budding from membranes of the endoplasmic reticulum and cytoplasmic vesicles. The particles in IBV-infected cells were similar in size and shape to those in strain 229E-infected cells but showed certain differences in internal structure. The evidence that the particles represent virions and the implications of these findings in the classification of this virus group are discussed.

Animals↗

Characterization of a cowpox-like orthopox virus which had caused a lethal infection in man.

In August 1990 an orthopox virus (OPV) had been isolated from a severe case of a generalized infection with lethal outcome in an immunosuppressed 18-year-old man. In this communication we present a detailed characterization of the causative virus strain. Based on distinct epitope configurations detected by various monoclonal antibodies the isolate could be differentiated from other OPV species and was classified as a cowpox virus (CP). This classification was confirmed by a species-specific PCR assay and by establishing physical maps for the restriction enzymes HindIII and XhoI. Based on serological data of neutralization assays, blocking-ELISAs and Western blotting analysis evidence is provided that the infection had been acquired from a stray cat.

Adolescent↗

AI-enabled viral genomics: from virus discovery to host prediction and emerging variant forecasting.

The rapid expansion of metagenomic sequencing has generated vast repositories of viral sequence data that far outpace our capacity to interpret them using conventional approaches. Highly divergent sequences, sparse functional annotation, and taxonomically uneven sampling present fundamental challenges for reference-dependent methods, which lose sensitivity precisely for novel and understudied viruses with high public health relevance. Artificial intelligence (AI) provides a new avenue to address these challenges by enabling predictive inference from viral genomes and proteins while reducing dependence on sequence similarity. In this Review, we discuss representative advances in AI for virus discovery, taxonomic classification and functional annotation, prediction of host range and zoonotic potential, and efforts toward forecasting emerging variants. These advances are transforming viral genomics from a largely descriptive discipline into one with increasing predictive capability. We also critically assess the major challenges that constrain current approaches, including the availability of high-quality and representative datasets, rigorous model evaluation, biological interpretability and responsible governance for increasingly capable AI models.

Artificial Intelligence↗

Identification and characterization of an adeno-associated virus integration site in CV-1 cells from the African green monkey.

Adeno-associated virus (AAV) is a classification given to a group of nonpathogenic, single-stranded DNA viruses known to reside latently in primates. During latency in humans, AAV type 2 (AAV2) preferentially integrates at a site on chromosome 19q13.3ter by targeting a sequence composed of an AAV Rep binding element (RBE), a spacer, and a nicking site. Here, we report the DNA sequence of an African green monkey AAV integration site isolated from CV-1 cells. Overall, it has 98% homology to the analogous human site, including identical spacer and nicking sequences. However, the simian RBE is expanded, having five perfect directly repeated GAGC tetramers. We carried out a number of in vitro and in vivo assays to determine the effect of this expanded RBE sequence on the Rep-RBE interaction and AAV targeted integration. Using electromobility shift assays it was demonstrated that AAV4 Rep68 bound the expanded RBE with a sixfold-greater affinity than the human RBE. To determine the basis for the affinity increase, DNase I protection and methylation interference (MI) assays were performed. Comparison of footprints on both the human and simian RBEs revealed nearly identical protection; however, MI analysis suggested greater interaction with the guanine nucleotides of the expanded RBE, thus providing a biochemical basis for the increased binding activity. In vivo, integration targeted to the simian RBE was demonstrated by PCR analysis of latently infected Cos-7 cells. Interestingly, the frequency of site-specific integration was twofold greater in Cos-7 cells than in HeLa cells. Overall, these experiments establish that the simian RBE, identified in CV-1 cells, functions analogously to the human RBE and provide further evidence for a developing model that proposes individual roles for the RBE and the spacer and nicking site elements.

Animals↗

[Virology of acute upper respiratory tract infections (author's transl)].

The virology of acute upper respiratory tract infections is presented in a survey. The first part includes general properties of viruses (structure, replication, maturation, classification), aetiological and immunological problems of viral infections. The different respiratory viruses and their clinical picture in particular are summarized in the second part.

Acute Disease↗