RESUMO
BACKGROUND: Porcine circovirus type 2 (PCV2) is a major emerging virus of porcine circovirus-associated disease (PCVAD), which has brought huge economic losses to the global pig industry. Pigs are well known as the natural reservoir of PCV2. Recently, many researchers have revealed PCV2 could infect many other mammals like mice, calves, minks, dogs and goats. In 2018, our laboratory has admitted six cases of raccoon dogs from Qinhuangdao city of China, which were characterized by inappetence, lethargy, depression, abortion, and sterility. RESULTS: At last, six raccoon dog-origin PCV2 strains were isolated in this study. Pairwise-sequence comparisons demonstrated that the six raccoon dog-origin PCV2 strains shared a nucleotide similarity of 92.1-99.8% among 40 PCV2 representative strains. Phylogenetic analysis indicated these PCV2 isolates belonged to Chinese epidemic genotypes PCV2b and PCV2d. And aborted or sterile symptom was significantly associated with PCV2 infection in raccoon dogs by the chi-square test (χ2 = 87.3, p < 0.001). The retrospective study revealed that raccoon dog-origin PCV2 strains shared 100% sequence similarity with the PCV2 stains isolated from pig farms around these raccoon dog farms, respectively. CONCLUSION: In this study, the first supported evidence of PCV2 prevalence in raccoon dog farms of China was documented. PCV2 may be one of the most significant causative agents resulting in the reproductive failure of farmed raccoon dogs, implying that PCV2 could transmit from pigs to raccoon dogs. That indicated that PCV2 cross-species transmission will be a serious threat to China's fur animal farming industry.
Assuntos
Infecções por Circoviridae/veterinária , Circovirus/genética , Doenças do Cão/virologia , Animais , Infecções por Circoviridae/virologia , Cães/virologia , Genoma Viral/genética , Filogenia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Análise de Sequência de DNARESUMO
Canine distemper virus (CDV) is a major pathogen not only in raccoon dogs but also in a variety of carnivorous animals, including domesticated animals, particularly if they have not been vaccinated. In this study, a wild-type strain of CDV was isolated from lung tissue from a raccoon dog kept at a fur farm in Jilin Province, China. Cytopathic effects typical of CDV infection were observed after three blind passages in Vero cells, yielding a virus titer of 10(4.6) TCID50/mL. Virus identification was carried out by RT-PCR, immunofluorescence, electron microscopy, and genome sequencing. The results showed that the isolated virus, termed the SY strain, corresponded to the Asia-1 genotype of CDV and has a genome of 15,690 nucleotides. This represents the first complete nucleotide sequence of a CDV strain circulating in raccoon dogs in China.
Assuntos
Vírus da Cinomose Canina/classificação , Vírus da Cinomose Canina/isolamento & purificação , Cinomose/virologia , Cães Guaxinins/virologia , Animais , China , Análise por Conglomerados , Efeito Citopatogênico Viral , Vírus da Cinomose Canina/genética , Imunofluorescência , Pulmão/virologia , Microscopia Eletrônica , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Análise de Sequência de DNA , Homologia de Sequência , Cultura de VírusRESUMO
OBJECTIVE: This study aimed to identify genes regulated by cyclin dependent kinases 5 (CDK5) that participate in hair pigmentation in mice. METHODS: The mRNA expression profiles of skin samples from CDK5-knockdown mice were constructed using high-throughput RNA sequencing and compared with those of wild-type mice. RESULTS: In total, 8,002 known genes were differentially expressed between CDK5-knockdown and wild-type mice. Of these, 3,658 were upregulated and 4,344 were downregulated in the skin of CDK5-knockdown mice. An additional 318 previously unknown genes were also differentially expressed, with 171 downregulated and 147 upregulated genes in the skin of CDK5-knockdown mice. Of the known genes expressed in mouse skin, 80 were associated with hair color, with 61 showing lower expression and 19 exhibiting higher expression in skin of CDK5-knockdown mice. Importantly, the expression of the tyrosinase-related protein 1 (TYRP1) and the calcium signaling pathway were also found to be regulated by CDK5, suggesting that pigmentation is regulated by CDK5 via the calcium signaling pathway and TYRP1. CONCLUSION: The transcriptome profiles obtained from the skin of CDK5-knockdown mice compared to wild-type mice provide a valuable resource to help understand the mechanism by which CDK5 regulates melanogenesis in mice and other animals.
RESUMO
A novel circovirus called porcine circovirus type 4 (PCV4) was recently detected in pigs suffering from severe clinical diseases in Hunan province, China. There are few reports on the origin and evolution of PCV4, although some researchers have conducted epidemiological investigations of PCV4 and found that PCV4 is widespread in pigs. Based on the previous study, we detected PCV2 in farmed foxes and raccoon dogs with reproductive failure. To explore whether the PCV4 genome also exists in fur animals, we detected 137 cases admitted from fur animal farms in Hebei China between 2015 and 2020, which were characterized by inappetence, lethargy, depression, abortion, and sterility. The overall infection rate of PCV4 was 23.36% (32/137), including 20.37% (22/108) for raccoon dogs, 18.75% (3/16) for foxes, and 53.85% (7/13) for minks. Finally, five raccoon dog-origin PCV4 strains and one fox-origin PCV4 strain were sequenced in our study, whose nucleotide identities with other representative PCV4 strains varied from 96.5% to 100%. Phylogenetic analysis based on the complete genomes of PCV4 strains indicated a close relationship with those of PCV4 strains identified from pigs. To our knowledge, this is the first study to detect PCV4 in fur animals. Interestingly, we also identified PCV4 in a mixed farm (feeding pigs and raccoon dogs at the same time). In summary, our findings extend the understanding of the molecular epidemiology of PCV4 and provide new evidence for its cross-species transmission.
RESUMO
Porcine Epidemic Diarrhea (PED) is a highly contagious intestinal disease which mostly caused by Porcine Epidemic Diarrhea Virus (PEDV). The PED has caused huge economic losses to the pig industry all over the world and a valid PEDV vaccine is needed to prevent the infection. In this study, we constructed expression plasmid based on the spike (S) gene of the epidemic PEDV strain. The recombinant eukaryotic S (Se) and prokaryotic S (Sp) subunit proteins were expressed and purified as vaccine antigens. We designed a new subunit vaccine based on S proteins, adjuvanted with layered double hydroxide (LDH). The results indicated that the LDH adjuvanted subunit vaccines induced a better immune effect in terms of antibody level and cellular immune response. In conclusion, this study showed a new design of a PEDV subunit vaccine with nanotechnology and demonstrated the potential for its clinical application.
Assuntos
Infecções por Coronavirus/imunologia , Hidróxidos/química , Imunidade , Nanopartículas/química , Vírus da Diarreia Epidêmica Suína/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Vacinas de Subunidades Antigênicas/imunologia , Adjuvantes de Vacinas/química , Animais , Anticorpos Antivirais , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Epidemias , Células HEK293 , Humanos , Nanotecnologia/métodos , Proteínas Recombinantes/imunologia , Suínos , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/virologia , Desenvolvimento de Vacinas/métodosRESUMO
From 2010, porcine epidemic diarrhea virus (PEDV) variants caused sequential outbreaks of disease in Asia and the United States. In this retrospective study, 49 complete spike (S) gene sequences were obtained from PEDV strains collected in China from 2014 to 2016. We observed that variant PEDV strains with novel insertions, deletions, and multiple S gene recombination types were present in China. In addition, mixed infections involving different variant strains were observed in some areas. Based on phylogenetic and recombination analyses, we determined that the newly emerged PEDV variants potentially originated via recombination between the earliest Chinese G1 genogroup strain, JS-2004-2 and earlier Korean pandemic strains. These findings provide important information for understanding ongoing PEDV outbreaks and suggest that novel variants make it more difficult to prevent PEDV infection.
Assuntos
Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/genética , Doenças dos Suínos/virologia , Animais , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Surtos de Doenças/veterinária , Genótipo , Filogenia , Estudos Retrospectivos , Suínos , Doenças dos Suínos/epidemiologiaRESUMO
Porcine circovirus type 2 (PCV2) is a causative agent of porcine circovirus-associated disease (PCVAD), which is a serious problem in the swine industry worldwide. In recent years, nonporcine-origin PCV2 has attracted more and more attention of the researchers. This study reported on the first identification of PCV2 in farmed foxes with reproductive failure. Three fox-origin PCV2 strains were successfully isolated, sequenced, and designated as FoxHB1, FoxHB2, and FoxHB3 respectively. Pairwise-sequence comparisons of the complete genomes revealed that three fox-origin PCV2 strains had nucleotide identities varied from 91.9% to 99.7% with representative strains of PCV2 different genotypes. Meanwhile, phylogenetic analysis based on complete genomes of 44 PCV2 strains indicated that the fox-origin PCV2 strains belonged to Chinese epidemic genotypes PCV2b and PCV2d. These results provided the first supported evidence that PCV2 could infect foxes, implying that the cross-species transmission of PCV2 would be a big threat to Chinese fur animal-bearing industry.
Assuntos
Infecções por Circoviridae/veterinária , Circovirus/genética , Circovirus/isolamento & purificação , Raposas/virologia , Animais , Infecções por Circoviridae/virologia , DNA Viral/genética , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Genoma Viral/genética , Genótipo , Filogenia , Reação em Cadeia da Polimerase/veterináriaRESUMO
Clathrin-mediated endocytosis is a mechanism used for the invasion of cells by a variety of viruses. Mortalin protein is involved in a variety of cellular functions and plays a role in viral infection. In this study, we found that mortalin significantly inhibited the replication of porcine epidemic diarrhea virus (PEDV) through restricting virus entry. Mechanistically, a biochemical interaction between the carboxyl terminus of mortalin and clathrin heavy chain (CLTC) was been found, and mortalin could induce CLTC degradation through the proteasomal pathway, thereby inhibiting the clathrin-mediated endocytosis of PEDV into host cells. In addition, artificial changes in mortalin expression affected the cell entry of transferrin, further confirming the above results. Finally, we confirmed that this host-mounted antiviral mechanism was broadly applicable to other viruses, such as vesicular stomatitis virus (VSV), rotavirus (RV), and transmissible gastroenteritis virus (TGEV), which use the same clathrin-mediated endocytic to entry. These results reveal a new function of mortalin in inhibiting endocytosis, and provide a novel strategy for treating PEDV infections.
Assuntos
Clatrina/metabolismo , Endocitose/fisiologia , Proteínas de Choque Térmico HSP70/metabolismo , Interações entre Hospedeiro e Microrganismos/fisiologia , Vírus da Diarreia Epidêmica Suína/fisiologia , Internalização do Vírus , Replicação Viral/fisiologia , Animais , Linhagem Celular , Chlorocebus aethiops , Regulação para Baixo/fisiologia , Inativação Gênica , Proteínas de Choque Térmico HSP70/genética , Células HeLa , Humanos , RNA Interferente Pequeno/metabolismo , Células VeroRESUMO
Porcine circovirus type 2 (PCV2), the primary causative agent of porcine circovirus-associated disease (PCVAD), causes severe economic losses to the pig industry in China since 2002. To investigate the molecular epidemic characteristics and genetic evolution of PCV2, 12 PCV2 isolates obtained from different pig farms with various clinical symptoms of PCVAD in Hebei, China from 2004 to 2014 were sequenced and analyzed. The phylogenetic analysis showed that the 12 isolates were divided into two distinct genotypes, PCV2b (7/12) and PCV2d (5/12), based on the sequences of either viral complete genome or open reading frame 2 (ORF2). Of the 7 PCV2b strains, 5 were isolated from 2004 to 2008 while all PCV2d were isolated from 2009 to 2014. This exhibited that PCV2b isolates were the most common before 2009 and then PCV2d isolates became predominant and widely distributed in pig farms. Sequence comparisons among total isolates indicated that the nucleotide identity ranged from 95.5% to 100% for complete genome and 93.1%-100% for ORF2. Compared with seven PCV2b isolates, there were thirteen amino-acid substitutions in the ORF2 region and one additional amino-acid K at this region terminal for five PCV2d isolates. The results suggest that a higher genetic variation and a distinct genotype shift occurred among the PCV2 isolates collected from 2004 to 2014 in Hebei.
Assuntos
Circovirus/genética , Variação Genética , Substituição de Aminoácidos , Animais , China , Infecções por Circoviridae/veterinária , Infecções por Circoviridae/virologia , Circovirus/classificação , Circovirus/isolamento & purificação , Genótipo , Fases de Leitura Aberta , Filogenia , Análise de Sequência de Proteína , Suínos , Doenças dos Suínos/virologiaRESUMO
Pseudorabies is an economically important disease in a variety ot animals caused by pseudorabies virus. Since 2011, pseudorabies outbreaks occurred in many regions of China. Related researches on this virus become a hot topic in virology and veterinary. One of the difficulties for pseudorabies prevention and control is innate immune evasion. Explorations on this issue are conducive to the development of vaccine and drugs. Therefore, this review summarized the recent research progress on the mechanisms of pseudorabies virus innate immune evasion. Theoretical direction was provided on effetive prevention and control of pseudorabies owing to this review.