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Host range relationships and the evolution of canine parvovirus.

Canine parvovirus (CPV) is an example of an unusual class of emerging virus-those that gain an altered host range through genetic variation and subsequently become widespread pathogens of their new and previously resistant host species. CPV was first detected in 1978 as the cause of new diseases in dogs throughout the world, when it rapidly spread throughout domestic populations, as well as becoming widespread in wild dogs. CPV was soon shown to be a variant of the long recognized feline panleukopenia virus (FPV), from which it differed in less than 1% at the nucleotide sequence level. Genetic analysis showed that virtually all of the biological differences between CPV and FPV, including the canine host range, were determined by three or four sequence differences in the viral capsid protein gene. Analysis of the atomic structures of the CPV and FPV capsids showed that the differences controlling host range were located within two different structural regions and were exposed on the capsid surface. The CPV which first emerged in 1978 appeared to be derived from a single ancestral sequence, which has allowed the ready analysis of the subsequent evolution of the virus in nature. Sequence analysis has also revealed that CPV strains have undergone a series of evolutionary selections in nature which have resulted in the global distribution of new virus variants. This was first seen in the global replacement between 1979 and 1981 of the original (1978) strain of the virus by a genetically and antigenically variant strain, and the subsequent widespread selection of other variants which have also become globally distributed. The genetic and antigenic variation in the virus strains was also correlated with changes in the host range of the virus, in particular in the ability to replicate in cats, and in canine host range differences seen in tissue culture cells.

Amino Acid Sequence↗

Host-specific effects of the korA-korB operon and oriT region on the maintenance of miniplasmid derivatives of broad host-range plasmid RK2.

Two genetic determinants are sufficient for small derivatives of broad host-range plasmid RK2 to replicate in different Gram-negative bacteria: trfA, which encodes a replication initiator, and oriV, the origin of replication. In this study, nonessential RK2 determinants in the region encoding oriT, the origin of conjugative transfer, and the korA-korB operon, whose products regulate trfA expression, were tested for their effects on the stability of mini-RK2 plasmids in eight different hosts. We found that determinants of both regions can substantially alter plasmid stability, but the effects are not uniform in all hosts. The results also indicate that the effects of the korA-korB operon extend beyond that of the regulation of trfA transcription. This study further illustrates the different requirements for stable plasmid maintenance in diverse bacteria and the ability of wild-type RK2 to adapt to a variety of intracellular environments. The data also provide further evidence for the involvement of different regions of RK2 for stable maintenance in various hosts.

Conjugation, Genetic↗

Construction of recombinants between molecular clones of murine retrovirus MCF 247 and Akv: determinant of an in vitro host range property that maps in the long terminal repeat.

The leukemogenic mink cell focus-forming (MCF) retroviruses such as MCF 247 have biological properties distinct from those of their ecotropic progenitors. Nucleotide sequences encoding portions of gp70, Prp15E, and the long terminal repeat differ between the two types of viruses. To investigate the role of each of these genetic elements in determining the biological properties of MCF viruses, we prepared infectious molecular clones of MCF 247 and generated a set of recombinants between these clones and a molecular clone of Akv, the ecotropic parent of MCF 247. Each molecular clone of MCF 247 was distinct. All the recombinants between Akv and MCF 247 yielded infectious virus upon transfection. Most interestingly, recombinants which contain the long terminal repeat of MCF 247 were found to have an in vitro host range property that has been correlated with high oncogenic activity and thymotropism of certain MCF isolates; namely, they plated with higher efficiency on SC-1 cells than on NFS mouse embryo cells. Nononcogenic MCF isolates showed a slight preference for NFS cells, whereas Akv virus plated with approximately equal efficiency on the two cell types.

Animals↗

The Trichoplusia ni granulovirus helicase is unable to support replication of Autographa californica multicapsid nucleopolyhedrovirus in cells and larvae of T. ni.

Baculovirus DNA helicases are essential for replication and are determinants of host range. Helicases of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Trichoplusia ni granulovirus (TnGV) differ markedly, although both viruses replicate efficiently in the cabbage looper, T. ni. To determine whether the TnGV helicase (P137) could support replication of AcMNPV in T. ni cells or larvae, the native AcMNPV helicase gene (p143) was disrupted and substituted with p137. P137 did not support replication when synthesized by the P143-deficient AcMNPV. Moreover, P137 did not inhibit AcMNPV replication when co-synthesized in the presence of the AcMNPV P143. These results suggest that although TnGV and AcMNPV replicate efficiently in T. ni, specific protein-protein or protein-DNA interactions between baculoviral helicases and viral-specific factors which form the replicase complex are required for virus replication. A novel and rapid method for disrupting AcMNPV genes in E. coli using the commercial Bac-to-Bac AcMNPV baculovirus expression vector is described.

Animals↗

Hormonal regulation of the gene for the type C ecotropic retrovirus receptor in rat liver cells.

The infectibility of the regenerating rat liver by ecotropic retroviruses was studied relative to the expression of the gene coding for the ecotropic retrovirus receptor (Ecor) that functions as a cationic amino acid transporter. It is known that the gene for the receptor is expressed in primary hepatocytes and hepatoma cells but is absent in adult liver cells. Isolation of a 2.85-kb cDNA for the rat Ecor suggested that the rat viral receptor is 97% homologous to the mouse viral receptor and that it contains the envelope-binding domain that determines the host range of ecotropic murine retroviruses. This explains the efficient infection of rat cells by ecotropic retroviruses. Since cell division is required for liver cells to be infected, we determined the susceptibility of the regenerating rat liver to infection at different time points after partial hepatectomy (0 to 24 h) in relation to the presence of receptor mRNA. Infection of the liver occurred only when the liver was exposed to virus 4 h after partial hepatectomy. This time course of infection paralleled expression of the gene for the Ecor, which was rapidly induced between 2 and 6 h during liver regeneration. However, expression of the dormant receptor gene in quiescent liver cells can be induced by insulin, dexamethasone, and arginine, indicating that cell division is not required for expression of the receptor gene in liver cells. A diet high in carbohydrate (low in protein) significantly increased the concentration of receptor mRNA in liver cells, indicating that hormones play a role in the regulation of expression of this gene in vivo. We conclude that the gene for the viral receptor is expressed in the regenerating and quiescent liver when the urea cycle enzymes are down regulated. The infection of the regenerating rat liver by ecotropic retroviruses at the time point of expression of the receptor gene supports the requirement of expression of this transporter for infection.

Amino Acid Sequence↗

Analysis of host range phenotypes of primate hepadnaviruses by in vitro infections of hepatitis D virus pseudotypes.

Hepatitis B virus (HBV) and woolly monkey hepatitis B virus (WMHBV) have natural host ranges that are limited to closely related species. The barrier for infection of primates seems to be at the adsorption and/or entry steps of the viral replication cycle, since a human hepatoma cell line is permissive for HBV and WMHBV replication following transfection of cloned DNA. We hypothesized that the HBV and WMHBV envelope proteins contain the principal viral determinants of host range. As previously shown by using the hepatitis D virus (HDV) system, recombinant HBV-HDV particles were infectious in chimpanzee as well as human hepatocytes. We extended the HDV system to include HDV particles pseudotyped with the WMHBV envelope. In agreement with the natural host ranges of HBV and WMHBV, in vitro infections demonstrated that HBV-HDV and WM-HDV particles preferentially infected human and spider monkey cells, respectively. Previous studies have implicated the pre-S1 region of the large (L) envelope protein in receptor binding and host range; therefore, recombinant HDV particles were pseudotyped with the hepadnaviral envelopes containing chimeric L proteins with the first 40 amino acids from the pre-S1 domain exchanged between HBV and WMHBV. Surprisingly, addition of the human amino terminus to the WMHBV L protein increased infectivity on spider monkey hepatocytes but did not increase infectivity for human hepatocytes. Based upon these data, we discuss the possibility that the L protein may be comprised of two domains that affect infectivity and that sequences downstream of residue 40 may influence host range and receptor binding or entry.

Amino Acid Sequence↗

Genetic analysis of tox+ and tox- bacteriophages of Corynebacterium diphtheriae.

A series of mutants derived from the temperate corynebacteriophages beta(tox+), gamma(tox-), and L(tox+) was isolated and characterized. In three-factor crosses between mutant beta phages the relative map order of the genetic markers determining extended host ranges (h and h') and loss of ability to lysogenize (c) was found to be h--c--h'. Recombination between markers was observed in matings between phage beta and the heteroimmune corynebacteriophages gamma and L. In such matings between heteroimmune phages the c markers of phages beta and gamma failed to segregate from the imm markers which determine the specificity of lysogenic immunity in these phages. The factor which directs the synthesis of diphtherial toxin during infection of appropriate corynebacterial hosts by toxinogenic corynebacteriophages is designated tox(+). It was possible to show that the tox(+) determinant of phage beta behaves as a single genetic element which occupies a position between the loci h and imm on the genetic map of this phage. Genetic recombination between mutants of phage beta occurred at very low frequencies in biparental matings performed by mixed infection of Corynebacterium diphtheriae C7(s)(-)(tox-). Considerably higher recombination frequencies were observed when lysogenic bacterial strains carrying one parental phage as prophage were induced by ultraviolet irradiation and then superinfected by the second parental phage. Maximal stimulation of genetic recombination between mutant beta phages was detected when superinfection followed ultraviolet irradiation of the lysogenic cells within a limited period of time. In matings between phages with incomplete genetic homology, the stimulation of recombination by ultraviolet radiation was much less effective.

Animals↗

A recombinant human parainfluenza virus type 3 (PIV3) in which the nucleocapsid N protein has been replaced by that of bovine PIV3 is attenuated in primates.

The shipping fever (SF) and Kansas (Ka) strains of bovine parainfluenza virus type 3 (BPIV3) are restricted in their replication in rhesus monkeys 100- to 1,000-fold compared to human parainfluenza virus type 3 (HPIV3), and the Ka strain also was shown to be attenuated in humans. To initiate an investigation of the genetic basis of the attenuation of BPIV3 in primates, we produced viable chimeric HPIV3 recombinants containing the nucleoprotein (N) open reading frame (ORF) from either BPIV3 Ka or SF in place of the HPIV3 N ORF. These chimeric recombinants were designated cKa-N and cSF-N, respectively. Remarkably, cKa-N and cSF-N grew to titers comparable to those of their HPIV3 and BPIV3 parents in LLC-MK2 monkey kidney and Madin-Darby bovine kidney cells. Thus, the heterologous nature of the N protein did not impede replication in vitro. However, cKa-N and cSF-N were each restricted in replication in rhesus monkeys to a similar extent as Ka and SF, respectively. This identified the BPIV3 N protein as a determinant of the host range restriction of BPIV3 in primates. These chimeras thus combine the antigenic determinants of HPIV3 with the host range restriction and attenuation phenotype of BPIV3. Despite their restricted replication in rhesus monkeys, the chimeric viruses induced a level of resistance to HPIV3 challenge in these animals which was indistinguishable from that conferred by immunization with HPIV3. The infectivity, attenuation, and immunogenicity of these BPIV3/HPIV3 chimeras suggest that the modified Jennerian approach described in the present report represents a novel method to design vaccines to protect against HPIV3-induced disease in humans.

Animals↗

Closing the ranks to attack by powdery mildew.

Powdery mildews are among the most common plant diseases, infecting over 650 monocot and over 9000 dicot species. Analysis in domesticated barley and wild Arabidopsis has begun to unravel the genetic and molecular frameworks underlying the mechanisms of susceptibility and resistance to these biotrophic fungal pathogens. This has revealed multiple pathways regulating host defense, some of which are also involved in determining the host range of the pathogen. Host-cell death and rapid cell-wall remodeling have emerged as frequent themes in powdery-mildew resistance. Several mutants have been isolated that might shed light on the enigma of susceptibility determinants in plants.

Fungi↗

The ability of two different Vibrio spp. bacteriophages to infect Vibrio harveyi, Vibrio cholerae and Vibrio mimicus.

AIMS: To determine the host range of the Vibrio harveyi myovirus-like bacteriophage (VHML) and the cholera toxin conversion bacteriophage (CTX Phi) within a range of Vibrio cholerae and V. mimicus and V. harveyi, V. cholerae and V. mimicus isolates respectively. METHODS AND RESULTS: Three V. harveyi, eight V. cholerae and five V. mimicus isolates were incubated with VHML and CTX Phi. Polymerase chain reaction (PCR) was used to determine the presence of VHML and CTX Phi in infected isolates. We demonstrated that it was possible to infect one isolate of V. cholerae (isolate ACM #2773/ATCC #14035) with VHML. This isolate successfully incorporated VHML into its genome as evident by positive PCR amplification of the sequence coding part of the tail sheath of VHML. Attempts to infect all other V. cholerae and V. mimicus isolates with VHML were unsuccessful. Attempts to infect V. cholerae non-01, V. harveyi and V. mimicus isolates with CTX Phi were unsuccessful. CONCLUSIONS: Bacteriophage infection is limited by bacteriophage-exclusion systems operating within bacterial strains and these systems appear to be highly selective. One system may allow the co-existence of one bacteriophage while excluding another. VHML appears to have a narrow host range which may be related to a common receptor protein in such strains. The lack of the vibrio pathogenicity island bacteriophage (VPI Phi) in the isolates used in this study may explain why infections with CTX Phi were unsuccessful. SIGNIFICANCE AND IMPACT OF THE STUDY: The current study has demonstrated that Vibrio spp. bacteriophages may infect other Vibrio spp.

Bacteriophages↗

Relationship between temperature optima and secreted protease activities of three Pythium species and pathogenicity toward plant and animal hosts.

The in vitro physiological characteristics of three species of Pythium (oomycetes) that utilize different food sources were compared with their ecological activities: P. insidiosum is a pathogen of mammals (including humans), P. graminicola infects the roots of graminaceous hosts, and P. grandisporangium is an enigmatic water mold isolated from mangrove leaves and marine algae. P. insidiosum and P. graminicola showed peak growth rates at 37 degrees C before complete inhibition of growth at 40 degrees C; P. grandisporangium grew fastest at 22 degrees C. Differences between the invasive pressures exerted by the hyphae of these microorganisms were not considered significant in relation to the substrates colonized by these water molds. All three species showed substantial secreted protease activity, producing three or more serine proteases with weights ranging from 24-38 kDa. Fastest growth rates were supported when collagen was supplied as the sole carbon source, and none of the species were able to grow on purified plant cell wall polysaccharides. The growth and nutritional characteristics of P. graminicola and P. grandisporangium bear little obvious relationship to the ecological niches that they inhabit. This highlights the caution necessary in extrapolating from laboratory analyses to the natural environment, and points to the potential importance of ecological opportunity in determining the host range and food source of certain microorganisms.

Animals↗

Rhizobium etli and Rhizobium gallicum nodulate common bean (Phaseolus vulgaris) in a traditionally managed milpa plot in Mexico: population genetics and biogeographic implications.

The stability of the genetic structure of rhizobial populations nodulating Phaseolus vulgaris cultivated in a traditionally managed milpa plot in Mexico was studied over three consecutive years. The set of molecular markers analyzed (including partial rrs, glnII, nifH, and nodB sequences), along with host range experiments, placed the isolates examined in Rhizobium etli bv. phaseoli and Rhizobium gallicum bv. gallicum. Cluster analysis of multilocus enzyme electrophoresis and plasmid profile data separated the two species and identified numerically dominant clones within each of them. Population genetic analyses showed that there was high genetic differentiation between the two species and that there was low intrapopulation differentiation of the species over the 3 years. The results of linkage disequilibrium analyses are consistent with an epidemic genetic structure for both species, with frequent genetic exchange taking place within conspecific populations but not between the R. etli and R. gallicum populations. A subsample of isolates was selected and used for 16S ribosomal DNA PCR-restriction fragment length polymorphism analysis, nifH copy number determination, and host range experiments. Plasmid profiles and nifH hybridization patterns also revealed the occurrence of lateral plasmid transfer among distinct multilocus genotypes within species but not between species. Both species were recovered from nodules of the same plants, indicating that mechanisms other than host, spatial, or temporal isolation may account for the genetic barrier between the species. The biogeographic implications of finding an R. gallicum bv. gallicum population nodulating common bean in America are discussed.

Agriculture↗

Use of type 1/type 2 chimeric polioviruses to study determinants of poliovirus type 1 neurovirulence in a mouse model.

We previously described the characteristics of a type 1/type 2 (PV-1/PV-2) chimeric poliovirus, v510, which contains the six amino acids specific for PV-2 in the B-C loop of VP1. This virus was found to be mouse-adapted, as PV-2 and in contrast with PV-1. Determinants of host range were studied in detail and are reported here. PV-1/PV-2 chimeras containing partial PV-1----PV-2 substitutions in the B-C loop of VP1 were obtained by making use of a mutagenesis cartridge on PV-1 cDNA. Analysis of mouse neurovirulence of these chimeras, when correlated with the three-dimensional structure of the v510 capsid, revealed that PV-2 residues important for mouse tropism are those which determine the particular conformation of the B-C loop of VP1 in v510. The mutation of the adenine residue at position 480 of the 5' noncoding region into a guanine residue has been shown to be an important determinant of PV-1 attenuation in monkeys. We show that introduction of this mutation in the v510 genome results in a virus which is partially attenuated for mice. This suggests that analysis of genomic determinants important for PV-1 neurovirulence could be carried out in a mouse model by making use of a mouse-adapted PV-1/PV-2 chimera.

Amino Acid Sequence↗

H5N1 Clade 2.3.4.4b Infections in Domestic Cats During an Avian Influenza Outbreak in Italy: Implications for One Health Surveillance.

BACKGROUND: H5Nx goose/Guangdong (Gs/GD) lineage highly pathogenic avian influenza (HPAI) viruses pose a significant public health threat due to their global spread, mutation accumulation, and expanding host range. The descendant clade 2.3.4.4b has been causing widespread infections in birds and increasing spillover events in mammals. METHODS: This report documents the first fatal case of highly pathogenic avian influenza virus (HPAIV) H5N1 clade 2.3.4.4b infection in a domestic cat in Italy, detected in early January 2025. The cat (CAT 1) resided on a backyard poultry farm experiencing a high pathogenicity avian influenza outbreak and succumbed rapidly following the onset of respiratory signs. A second exposed cat (CAT 2) developed clinical disease without fatal outcome. Comprehensive outbreak investigations were conducted, including pathological, serological, molecular analyses, and genomic characterization. RESULTS: Pathological examination of CAT 1 revealed acute necrotizing bronchointerstitial pneumonia, non-suppurative meningoencephalitis, and disseminated foci of hepatic necrosis. Interestingly, the PB2-E627K mutation associated with mammalian virus adaptation was observed in the feline viral isolate compared to avian isolates. Such polymerase complex mutations are key determinants of host range and increase pathogenicity in mammals. CAT 2 from the same farm tested negative for AIV genome detection but subsequently seroconverted for antibodies against NPA, H5, and N1. CONCLUSION: Sharing these findings is crucial for surveillance aimed at enabling early identification of increased risks to human and animal health, preventing cross-species viral transmission and mitigating the risk of potential spillover events.

Animals↗

Two novel adeno-associated viruses from cynomolgus monkey: pseudotyping characterization of capsid protein.

We demonstrated the presence of two adeno-associated viruses (AAVs), designated AAV10 and AAV11, in cynomolgus monkeys by isolating and sequencing the entire viral coding regions from the monkey DNA. AAV10 and AAV11 capsid proteins shared 84% and 65%, respectively, of amino acids with AAV2. A phylogenetic analysis of AAV capsid proteins showed that AAV10 and AAV11 resembled most AAV8 and AAV4, respectively. To characterize the capsid protein, we pseudotyped an AAV2 vector with the monkey AAV capsid proteins and examined the resulting pseudotypes AAV2/10 and AAV2/11, in comparison with the AAV2 vector, for their host ranges in cell lines and tissue tropism in mice. AAV2/10 and AAV2/11 transduced primate cells less efficiently than AAV2. Whereas AAV2 transduced undifferentiated C2C12 mouse myoblasts more efficiently than differentiated ones, AAV2/10 and AAV2/11 transduced the undifferentiated myoblasts less efficiently than differentiated ones. Three weeks after injection to the muscle of the hind legs, AAV2/10 and AAV2 induced transgene expression similarly, but AAV2/11 did not transduce the skeletal muscle. Six weeks after systemic administration, transduced vector DNA was detected by PCR in the liver and spleen of mice inoculated with AAV2, in the liver, heart, muscle, lung, kidney, and uterus of mice with AAV2/10, and the muscle, kidney, spleen, lung, heart, and stomach of mice with AAV2/11. Mouse antisera against capsid protein VP2 of the three AAVs neutralized the respective vector particles in a type-specific manner. The results indicate that AAV10 and AAV11 capsid proteins, which are antigenically distinct from each other and AAV2, are likely to determine their host ranges and tissue tropism that are different from AAV2s, suggesting that cynomolgus AAVs could provide a broader choice of pseudotype AAV vectors for gene therapy.

Amino Acid Sequence↗

Role of nonstructural proteins 3A and 3B in host range and pathogenicity of foot-and-mouth disease virus.

The genome of foot-and-mouth disease virus (FMDV) differs from that of other picornaviruses in that it encodes a larger 3A protein (>50% longer than poliovirus 3A), as well as three copies of protein 3B (also known as VPg). Previous studies have shown that a deletion of amino acids 93 to 102 of the 153-codon 3A protein is associated with an inability of a Taiwanese strain of FMDV (O/TAW/97) to cause disease in bovines. Recently, an Asian virus with a second 3A deletion (amino acids 133 to 143) has also been detected (N. J. Knowles et al., J. Virol. 75:1551-1556, 2001). Genetically engineered viruses harboring the amino acids 93 to 102 or 133 to 143 grew well in porcine cells but replicated poorly in bovine cells, whereas a genetically engineered derivative of the O/TAW/97 virus expressing a full-length 3A (strain A12) grew well in both cell types. Interestingly, a virus with a deletion spanning amino acid 93 to 144 also grew well in porcine cells and caused disease in swine. Further, genetically engineered viruses containing only a single copy of VPg were readily recovered with the full-length 3A, the deleted 3A (amino acids 93 to 102), or the "super" deleted forms of 3A (missing amino acids 93 to 144). All of the single-VPg viruses were attenuated in porcine cells and replicated poorly in bovine cells. The single-VPg viruses produced a mild disease in swine, indicating that the VPg copy number is an important determinant of host range and virulence. The association of VPg copy number with increased virulence in vivo may help to explain why all naturally occurring FMDVs have retained three copies of VPg.

Animals↗

Systemic movement of an RNA plant virus determined by a point substitution in a 5' leader sequence.

The ability of viruses to move through infected plants is an important determinant of host range and pathogenicity. We have investigated the genetic basis for the inability of the Type strain of barley stripe mosaic hordeivirus to undergo long-range systemic movement in the tobacco Nicotiana benthamiana. We show that, in this model system, a short open reading frame in the 5' leader of the smallest viral genomic RNA prevents long-range vascular movement. As predicted by the ribosome scanning model, the leader open reading frame decreases the efficiency with which the 5'-proximal gene is translated in vitro. Thus, systemic pathogenicity in this system may be determined by the efficiency of translation of a viral gene in vivo and is not determined by the primary sequence of the encoded protein.

Amino Acid Sequence↗

Mapping of host range restriction of the rakkyo strain of tobacco mosaic virus in Nicotiana tabacum cv. bright yellow.

The rakkyo strain (TMV-R) and the common strain (TMV-U1) of tobacco mosaic virus exhibit distinct host range differences. TMV-R infects rakkyo plants, a monocot host that TMV-U1 is unable to infect. However, TMV-R causes only latent infection in Nicotiana tabacum cv. Bright Yellow (BY) in inoculated leaves, whereas TMV-U1 infects BY systemically and induces mosaic symptoms. Complete nucleotide sequencing of the TMV-R genomic RNA revealed amino acid changes in the 130K/ 180K replicase proteins, the 30K protein, and the coat protein and nucleotide changes in the 5' and 3' noncoding regions compared to TMV-U1. To identify viral components involved in determination of differences in host range, we have mapped determinants for the differential infection phenotype in BY plants by constructing chimeric viruses between the two strains in the present study. Examination of the infection phenotypes of the 14 constructed chimeric viruses in BY showed that determinants defining the differential infection phenotype in BY reside in the 130K/180K replicase proteins and the 3' noncoding region. Cognate combination of the 130K/180K replicase proteins and the 3' noncoding region of TMV-U1 origin is required to produce systemic infection in BY plants.

DNA-Directed DNA Polymerase↗