Establishment and application of multiplex real-time PCR for simultaneous detection of four viruses associated with porcine reproductive failure

Frontiers in Microbiology, Год журнала: 2023, Номер 14

Опубликована: Фев. 9, 2023

Many pathogens cause reproductive failure in sows suffering a broad spectrum of sequelae, including abortions, stillbirth, mummification, embryonic death, and infertility. Although various detection methods, such as polymerase chain reaction (PCR) real-time PCR, have been widely used for molecular diagnosis, mainly single pathogen. In this study, we developed multiplex PCR method the simultaneous porcine circovirus type 2 (PCV2), 3 (PCV3), parvovirus (PPV) pseudorabies virus (PRV) associated with failure. The R values standard curve PCV2, PCV3, PPV, PRV reached to 0.996, 0.997, 0.998, respectively. Importantly, limit (LoD) PRV, were 1, 10, 10 copies/reaction, Meanwhile, specificity test results indicated that is specific these four target does not react other pathogens, classical swine fever virus, respiratory syndrome epidemic diarrhea virus. Besides, had good repeatability coefficients variation intra- inter-assay less than 2%. Finally, approach was further evaluated by 315 clinical samples its practicality field. positive rates 66.67% (210/315), 8.57% (27/315), 8.89% (28/315), 4.13% (13/315), overall co-infection two or more 13.65% (43/315). Therefore, provides an accurate sensitive identification those underlying DNA viruses among potential pathogenic agents, allowing it be applied diagnostics, surveillance, epidemiology.

Язык: Английский

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Epizootiological surveillance of porcine circoviruses in free-ranging wild boars in China

Virologica Sinica, Год журнала: 2023, Номер 38(5), С. 663 - 670

Опубликована: Сен. 1, 2023

Four species of porcine circoviruses (PCV1-4) have been reported to circulate in Chinese domestic pigs, while the epizootiology these viruses free-ranging wild boars China remains unknown. In this study, tissue and serum samples collected from diseased or apparently healthy between 2018 2020 19 regions were tested for prevalence PCV1-4 infections. Positive rates PCV1, PCV2, PCV3 DNA 1.6% (4/247), 58.3% (144/247), 10.9% (27/247) respectively, with none positive PCV4. Sequence analysis viral genome showed that four PCV1 strains distributed Hunan Inner Mongolia shared 97.5%-99.6% sequence identity global reference strains. Comparison ORF2 gene sequences 80 PCV2 widely 18 79.5%-100% pigs boars, grouped into PCV2a (7), PCV2b (31) PCV2d (42). For PCV3, 17 sequenced 97.2%-100% nucleotide at genomic level could be divided PCV3a (3), PCV3b (2) PCV3c (12) based on phylogeny sequences. Serological data revealed antibody against 11.4% (19/167) 53.9% (90/167) respectively. The obtained study improved our understanding about epidemiological situations PCVs infection will valuable prevention control diseases caused by infection.

Язык: Английский

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Plasma Virome Reveals Blooms and Transmission of Anellovirus in Intravenous Drug Users with HIV-1, HCV, and/or HBV Infections

Microbiology Spectrum, Год журнала: 2022, Номер 10(4)

Опубликована: Июнь 27, 2022

Intravenous drug users (IDUs) are a high-risk group for HIV-1, hepatitis C virus (HCV), and B (HBV) infections, which the leading causes of death in IDUs. However, plasma virome IDUs how it is influenced by above viral infections remain unclear. Using metagenomics, we determined its association with HCV, and/or HBV infections. Compared healthy individuals, especially those major had higher abundance diversity. Anelloviridae dominated virome. Coinfections multiple anelloviruses were common, from same genus tended to coexist together. In this study, 4,487 anellovirus ORF1 sequences identified, including 1,620 (36.1%) less than 69% identity any known sequences, tripled current number. controls (HC), more observed neg-IDUs, further expanded sequence number IDUs, was characterized emergence novel divergent taxons blooms resident anelloviruses. Pegivirus mainly identified infected Five main pegivirus transmission clusters (TCs) phylogenetic analysis, suggesting link. Similar profiles within TC, anellome among Our data suggested that suffered burden anelloviruses, associated Blooms unprecedented diversity highlighted active evolution replication blood circulation, an uncharacterized role may engage host. IMPORTANCE Virome immune status determines or influences disease progression through both pathogenic viruses. Increased indicated suboptimal high infection risks these population. sensitive response other addition, cluster analysis revealed link individuals links shared similar profiles. In-depth monitoring populations not only needed surveillance emerging viruses networks neglected bloodborne viruses, but also important better understanding commensal their system.

Язык: Английский

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Vaccination Failures in Pigs—The Impact of Chosen Factors on the Immunisation Efficacy

Vaccines, Год журнала: 2023, Номер 11(2), С. 230 - 230

Опубликована: Янв. 19, 2023

Infectious diseases that often lead to economic losses still pose a severe problem in the pig production sector. Because of increasing restrictions on antibiotic usage, vaccines may become one major approaches controlling infectious diseases; much research has proved they could be very efficient. Nevertheless, during their life, pigs are exposed various factors can interfere with vaccination efficacy. Therefore, present paper, we reviewed influence chosen immunisation process, such as stress, faecal microbiota, host genetics, presence MDAs, infections immunosuppressive pathogens, and treatment antibiotics mycotoxins. Many them turned out have an adverse impact vaccine

Язык: Английский

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A Diverse Virome Is Identified in Parasitic Flatworms of Domestic Animals in Xinjiang, China

Microbiology Spectrum, Год журнала: 2023, Номер 11(3)

Опубликована: Апрель 12, 2023

Parasitic flatworms infect diverse vertebrates and are major threats to animal even human health; however, little is known about the virome of these lower life forms. Using viral metagenomic sequencing, we characterized parasitic collected from domestic animals, including Dicrocoelium lanceatum Taenia hydatigena, Echinococcus granulosus sensu stricto multilocularis. Seven three different viruses were discovered D. T. respectively, no sequences found in adult tapeworms protoscoleces E. Two out five flatworm species carry viruses, showing a host specificity viruses. These belong Parvoviridae, Circoviridae, unclassified circular, Rep-encoding single-stranded (CRESS) DNA virus, Rhabdoviridae, Endornaviridae, RNA The presence multiple highly divergent especially those that cluster with marine implies deep evolutionary history parasite-associated In addition, high identity common pathogens dogs, canine circovirus parvovirus 2. parasites they may but does not completely exclude possibility contamination intestinal contents. Furthermore, demonstrated certain such as CRESS virus integrate into genome their host. Our results expand knowledge diversity important highlighting need for further investigations prevalence among other key animals. IMPORTANCE Characterizing unveiling diversity, evolution, ecology will help understand "Russian doll" pattern parasites, data indicate present specific flatworms, have an ancient currently circulating parasite-infected also raise question whether acquire and/or some transmission potential through study virus-parasite-host interactions, influence infection on cycle parasite, well its fitness pathogenicity host, could find new strategies prevent control diseases.

Язык: Английский

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The first dog-origin porcine circovirus type 4 complete genomic sequence have high homology with that of pig-derived strains

Frontiers in Microbiology, Год журнала: 2023, Номер 14

Опубликована: Фев. 24, 2023

Introduction: Porcine circovirus 4 (PCV4) was discovered in 2019 and then proved to be pathogenic piglets. Nevertheless, few studies were currently available about PCV4 infection species other than pigs there is no information the prevalence of dogs. Methods: To fill this gap, 264 dog samples collected from animal hospitals Southwest China 2021 2022 screened for PCV4. Moreover, complete genome one strain (SCABTC-Dog2022) obtained successfully shared a high identity (97.9-99.0%) with strains derived pigs, dairy cows, raccoon dogs foxes. The SCABTC-Dog2022 analyzed together 51 reference sequences. Results Discussion: detected results showed low percentage PCV-4 DNA (1.14%, 3/264), indicating that could identified southwest China. Phylogenetic tree clustered closed relative geographically coherent branch four provinces (Sichuan, Fujian, Hunan Inner Mongolia) our knowledge, it first detection globally. association between status clinical syndromes deserves additional investigations.

Язык: Английский

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Virome of wild rats (Rattus norvegicus) captured far from pig farms in Jiangsu province of China reveals novel porcine circovirus type 2d (PCV2d) sequences

Virology Journal, Год журнала: 2023, Номер 20(1)

Опубликована: Март 9, 2023

Abstract Background Porcine circovirus type 2 (PCV2) has caused great economic losses in the global pig industry. There have been published records of wild rats acting as reservoirs PCV2 (only PCV2a and PCV2b), but almost all which were related to PCV2-infected swine herds. Results In this study, we carried out detection, amplification, characterization novel strains that captured far from farms. Nested PCR assay demonstrated kidney, heart, lung, liver, pancreas, large small intestines screened positive for PCV2. We subsequently sequenced two full genomes sample pools, designated js2021-Rt001 js2021-Rt002. Genome sequence analysis indicated they had highest similarity nucleotide sequences porcine-origin isolates Vietnam. Phylogenetically, js2021-Rt002 a part PCV2d genotype cluster, is predominant circulating worldwide recent years. The antibody recognition regions, immunodominant decoy epitope, heparin sulfate binding motif complete genome coincided with those previously reported. Conclusions Our research reported genomic (js2021-Rt001 js2021-Rt002) provided first supported evidence could naturally infect China. However, whether newly identified potential nature vertical horizontal transmission or inter-species jumping between pigs needs further research.

Язык: Английский

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Live attenuated Salmonella enterica serovar Choleraesuis vector delivering a virus-like particles induces a protective immune response against porcine circovirus type 2 in mice

Vaccine, Год журнала: 2022, Номер 40(33), С. 4732 - 4741

Опубликована: Июнь 27, 2022

Язык: Английский

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Detection and Complete Genome Analysis of Porcine Circovirus 2 (PCV2) and an Unclassified CRESS DNA Virus from Diarrheic Pigs in the Dominican Republic: First Evidence for Predominance of PCV2d from the Caribbean Region

Viruses, Год журнала: 2022, Номер 14(8), С. 1799 - 1799

Опубликована: Авг. 17, 2022

We report here high rates (47.5%, 48/101) of detection porcine circovirus 2 (PCV2) in diarrheic pigs from three pig farms the Dominican Republic. Seventeen PCV2 positive samples, representing farms, different age groups and sampling periods (2020-2021), were amplified for complete genome. Based on analysis open reading frame genome sequences, 17 strains assigned to PCV2d genotype. Significant differences observed between vaccinated (20% (10/50)) unvaccinated (62.5% (10/16) 80% (28/35)) corroborating previous observations that PCV2a-based vaccines confer protection against heterologous genotypes. The present study is first molecular characterization Republic, warranting large-scale epidemiological studies backyard systems across country. For time, was identified as predominant genotype a Caribbean region, suggesting shift PCV2b might be happening which mirrored current scenario many other parts world. Besides PCV2, we also pigeon circovirus-like virus, circular Replication-associated protein (Rep)-encoding single-stranded (CRESS) DNA characterized CRESS virus shared similar genomic organization related unclassified CRESSV2 viruses (belonging Order Cirlivirales) feces Hungary, indicating are circulating among geographical regions, further epidemiology biology these novel viruses.

Язык: Английский

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Genetic Diversity of Porcine Circovirus 2 Subtypes in Wild Boar and Domestic Pigs in Ukraine

Опубликована: Апрель 7, 2022

Porcine circovirus type 2 (PCV2) is responsible for a number of porcine associated diseases (PCAD) that can severely impact domestic pig herds. For non-enveloped virus with small genome (1.7 kb ssDNA), PCV2 remarkably diverse, 8 subtypes (a-h). New spread through migration wild boars, which are thought to infect pigs and further the trade. Despite large swine population, diversity in Ukraine has been undersampled, few sequences reported past decade. To gain deeper understanding subtype Ukraine, samples blood serum were collected from boars (n = 107) hunted during November-December 2012 hunting season. We found 34/107 (31.8%) prevalence by diagnostic PCR. pigs, liver 16) commercial market near Kharkiv 2019, 6/16 (37%) positive PCV2. sequenced genotyping locus ORF2, gene encoding viral capsid (Cp), 11 boar 6 using an Oxford Nanopore MinION device. Of 17 resolved subtypes, b was most common (10/11, 91%), while infected d. also detected b/d b/a co-infections respectively, f Poltava first time Ukraine. Building maximum likelihood phylogeny, we identified sublineage infections both swine, suggesting possible epizootic cluster ecological interaction northeastern

Язык: Английский

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