Isolation, identification and pathogenic characteristics of tick-derived parainfluenza virus 5 in northeast China.

The number of parainfluenza virus 5 (PIV5) infection cases has increased worldwide over the past six decades; however, factors underlying this increase remain unclear. PIV5 has been emerging or re-emerging in humans and animal species. To date, no information is yet available regarding PIV5 infection in arthropod ticks. Here, we successfully isolated tick-derived PIV5 from the Ixodes persulcatus species designated as HLJ/Tick/2019 in Heilongjiang, China. Phylogenetic analysis revealed that the tick-derived PIV5 is closely related to subclade 2.2.6, which has become the dominant subtype prevalent in dogs, pigs and wildlife across China. Further experiments to understand the importance of this virus as an infectious vector revealed that a ferret animal model experimentally infected with Tick/HLJ/2019 via the oronasal and ocular inoculation routes developed moderate respiratory distress with pneumonia and neurologic tissue damage from inflammation for the first time. Further surveillance of PIV5 in vectors of viral transmission is necessary to enhance our knowledge of its ecology in reservoirs and facilitate the control of re-emerging diseases.

Novel Mammalian orthorubulavirus 5 Discovered as Accidental Cell Culture Contaminant.

A distinct Russian Mammalian orthorubulavirus 5 (PIV5) was detected in cell culture exhibiting cytopathic effect and hypothesized to be contaminated by a scientist with respiratory symptoms. The identification of the divergent strain indicated a lack of knowledge on the diversity of PIV5 strains and calls for surveillance of global PIV5 strains.

Complete Genome Sequence of Parainfluenza Virus 5 (PIV5) from a Sunda Pangolin (Manis javanica) in China.

We report here the complete genome sequence of the parainfluenza virus PIV5-GD18 strain, isolated from a wild Sunda pangolin (Manis javanica) in China in 2017. It was 15,246 nucleotides with four nucleotides substitutions, which resulted in four changes of amino acid that were found only in PIV5-GD18, which further broadens the PIV5 infection host spectrum and will aid in our understanding of the complete genome sequence of PIV5 in different hosts.

Isolation and molecular characterization of parainfluenza virus 5 in diarrhea-affected piglets in China.

Recently, parainfluenza virus 5 (PIV5) infection has been increasingly reported in mammals. In this study, five PIV5 strains were isolated from diarrhea-affected piglets from four provinces or municipalities in China. An F-gene-based phylogenetic tree indicated that the five isolated strains were closely related to the PIV5 strain ZJQ-221 from a lesser panda in China, and the PIV5 strain 1168-1 from a dog in South Korea. The new isolates differed genetically from other pig, calf, rhesus macaque kidney cells, human, and dog PIV5 reference strains. Our study reveals the presence of PIV5 in intestinal tissue samples collected from diarrhea-affected piglets, and provides novel information regarding the epidemiology and tissue tropism of PIV5.

Isolation and genomic characterization of a canine parainfluenza virus type 5 strain in China.

A canine parainfluenza virus type 5 strain was isolated from a lung sample from a diseased dog. The genome sequene of this isolate, named HeN0718, was determined and compared tho those of other previously reported canine parainfluenza viruses. Unlike previously reported viruses, the HeN0718 strain contained several nucleotide mutations in the SH gene that led to a frame shift in the open reading frame. Phylogenetic analysis based on the complete virus genome and the P, F, and HN genes showed that HeN0718 was genetically closest to D277, a Korean strain that was isolated in 2008.

First complete genome sequence of parainfluenza virus 5 isolated from lesser panda.

Parainfluenza virus 5 (PIV5) is widespread in mammals and humans. Up to now, there is little information about PIV5 infection in lesser pandas. In this study, a PIV5 variant (named ZJQ-221) was isolated from a lesser panda with respiratory disease in Guangzhou zoo in Guangdong province, southern China. The full-length genome of ZJQ-221 was found to be 15,246 nucleotides and consisted of seven non-overlapping genes encoding eight proteins (i.e., NP, V, P, M, F, SH, HN and L). Sequence alignment and genetic analysis revealed that ZJQ-221 shared a close relationship with a PIV5 strain of canine-origin (1168-1) from South Korea. The findings of this study confirm the presence of PIV5 in lesser panda and indicate this mammal as a possible natural reservoir. Furthermore they highlight the urgent need to strengthen viral surveillance and control of PIV5 in zoo animals.

Molecular characteristics of canine parainfluenza viruses type 5 (CPIV-5) isolated in Korea.

Three canine parainfluenza viruses type 5 (CPIV-5) were isolated from lung tissues of 3 Korean dogs with mild pneumonia between 2008 and 2009. The isolates were fully sequenced and compared with published reference sequences. The size of the genome was 15 246 nucleotides long and no remarkable differences were found when compared with previously published reference sequences. In phylogenetic analysis based on the F and P genes, parainfluenza virus 5 (PIV-5) strains were divided into at least 3 subgroups. Three CPIV-5 strains were clustered with CPIV-5 T1, H22 and 78524 strains. All PIV-5 strains were independent of the host species, geographical distribution, and the isolated period.

Trois virus parainfluenza canins de type 5 (CPIV-5) furent isolés entre 2008 et 2009 à partir de tissu pulmonaire de trois chiens coréens souffrant de pneumonie légère. Les isolats furent complètement séquencés et comparés à des séquences de référence publiées. La taille du génome était de 15 246 nucléotides de longueur et aucune différence notable ne fut trouvée lorsque comparé avec les séquences de référence. Lors d'analyses phylogénétiques basées sur les gènes F et P, les souches de virus parainfluenza de type 5 (PIV-5) furent séparées en au moins trois sous-groupes. Les trois souches de CPIV-5 étaient regroupées avec les souches de CPIV-5 T1, H22 et 78524. Toutes les souches de PIV-5 étaient indépendantes de l'espèce hôte, de la distribution géographique et de la période à laquelle elles furent isolées.(Traduit par Docteur Serge Messier).

Stability of the parainfluenza virus 5 genome revealed by deep sequencing of strains isolated from different hosts and following passage in cell culture.

UNLABELLED: The strain diversity of a rubulavirus, parainfluenza virus 5 (PIV5), was investigated by comparing 11 newly determined and 6 previously published genome sequences. These sequences represent 15 PIV5 strains, of which 6 were isolated from humans, 1 was from monkeys, 2 were from pigs, and 6 were from dogs. Strain diversity is remarkably low, regardless of host, year of isolation, or geographical origin; a total of 7.8% of nucleotides are variable, and the average pairwise difference between strains is 2.1%. Variation is distributed unevenly across the PIV5 genome, but no convincing evidence of selection for antibody-mediated evasion in hemagglutinin-neuraminidase was found. The finding that some canine and porcine, but not primate, strains are mutated in the SH gene, and do not produce SH, raised the possibility that dogs (or pigs) may not be the natural host of PIV5. The genetic stability of PIV5 was also demonstrated during serial passage of one strain (W3) in Vero cells at a high multiplicity of infection, under conditions of competition with large proportions of defective interfering genomes. A similar observation was made for a strain W3 mutant (PIV5VΔC) lacking V gene function, in which the dominant changes were related to pseudoreversion in this gene. The mutations detected in PIV5VΔC during pseudoreversion, and also those characterizing the SH gene in canine and porcine strains, predominantly involved U-to-C transitions. This suggests an important role for biased hypermutation via an adenosine deaminase, RNA-specific (ADAR)-like activity. IMPORTANCE: Here we report the sequence variation of 16 different isolates of parainfluenza virus 5 (PIV5) that were isolated from a number of species, including humans, monkeys, dogs, and pigs, over 4 decades. Surprisingly, strain diversity was remarkably low, regardless of host, year of isolation, or geographical origin. Variation was distributed unevenly across the PIV5 genome, but no convincing evidence of immune or host selection was found. This overall genome stability of PIV5 was also observed when the virus was grown in the laboratory, and the genome stayed remarkably constant even during the selection of virus mutants. Some of the canine isolates had lost their ability to encode one of the viral proteins, termed SH, suggesting that although PIV5 commonly infects dogs, dogs may not be the natural host for PIV5.

Characterization in vitro and in vivo of a novel porcine parainfluenza virus 5 isolate in Korea.

A novel porcine parainfluenza 5 (pPIV5), KNU-11, in the genus Rubulavirus of the subfamily Paramyxovirinae, was isolated from the lung of a piglet in Korea in 2011. To understand the importance of this virus as an infectious agent, in vitro and in vivo characteristics of KNU-11 virus was investigated. KNU-11 was remarkably cytopathogenic, showing distinct cell rounding and clumping evident in porcine alveolar macrophage (PAM), porcine kidney (PK-15), and swine testicle (ST) cells within 12h postinfection and capable of hemagglutinating guinea pig red blood cells. Interestingly, this cytopathology was found to be absent in cell lines from other mammalian species. To evaluate the in vitro immunity of the pPIV5 isolate, we sought to explore alteration of inflammatory cytokine and chemokine expression in PAM cells infected with KNU-11 by using quantitative real-time RT-PCR. Most cytokine and chemokine genes including type 1 interferons (IFN-α/ß) and IFN-related antiviral genes were found to be significantly elevated in KNU-11 virus-infected PAM cells. A serum neutralization test-based serosurvey demonstrated that neutralizing antibodies against KNU-11 are readily detected in domestic swine populations, suggesting high prevalence of pPIV5 in Korean pig farms. Animal studies showed that KNU-11 fails to establish an acute respiratory illness, indicating that pPIV5 is non- or very mildly pathogenic to pigs.