Porcine circovirus 2 and 3 in wild boars in Southern Brazil

ABSTRACT: Porcine circovirus 2 (PCV2) has a considerable economic impact on the pork industry worldwide for more than two decades. In 2016, a new circovirus, porcine circovirus 3 (PCV3), was described; since then, it has been reported to be associated with diseased or even in clinically healthy swine in several countries. Considering the importance of wild boars as reservoirs of swine pathogens and the extensive distribution of these animals in Rio Grande do Sul and throughout the national territory, we searched for PCV2 and PCV3 in twenty-six wild boars coupled with necropsy and histologic examination of the sampled animals. Using PCR, 182 tissue samples were analyzed, including the heart, kidneys, liver, lung, lymph nodes, spleen, and tonsils. PCV2 and PCV3 were detected in 57.7% (15/26) and 15.4% (4/26) of wild boars, respectively. Furthermore, co-infection with PCV2 and PCV3 was detected in one of these animals, with PCV2 or PCV3 DNA detection in multiple organs. Histological examination showed mild to moderate and multifocal lymphoplasmacytic interstitial nephritis distributed randomly throughout the renal cortex, apparently unrelated to PCV2 or PCV3 detection. The wild boar population in Brazil is extensive, indicating the presence of a larger number of swine pathogen hosts. In the present study, more than half of the wild boars harbored PCV2; and although less frequently, PCV3 was also detected. Therefore, free-living wild boars can serve as reservoirs of swine circoviruses in southern Brazil.

Comparative Genome Analysis of Streptococcus suis Serotype 9 Isolates from China, The Netherland, and the U.K.

Streptococcus suis (S. suis) is an important swine pathogen and an emerging zoonotic agent worldwide. Serotype 9 is the most prevalent serotype in several European countries but it is relatively rare in China. In this study, through the investigation of the serotypes of 279 S. suis strains isolated from China from 2015 to 2017, it was found that serotype 9 is the second most prevalent serotype (43 out of 279), behind serotype 2 (83 out of 279). Next, the 43 serotype 9 isolates were sequenced and compared with those from the Netherland (28) and the U.K. (eight). For the purpose of comparison, the strain D12 (GCA_000231905), which has completed genome sequences, was also incorporated. Phylogenetic tree analysis showed that the strains from China and the U.K. were heterogeneous. In contrast, all but one from the Netherland belonged to the same clade. The dominant clades of Chinese strains (33) and strains from the Netherland (27) were very similar. Both of them may have originated from the same strain about 70 years ago. Then, the distributions of virulence-associated genes and antibiotic resistance genes among different clades and sources were analyzed. By comparison, strains from the Netherland carried more virulence-associated genes and those from the U.K. had more antibiotic resistance genes. Additionally, some virulence-associated genes (salK and salR) and antibiotic resistance genes (lincomycin and spectinomycin) existed only in several Chinese strains. In conclusion, our data displayed the population characteristics and differences of S. suis serotype 9 between China and Europe, suggesting that they have taken different evolutionary paths.

Neutrophils in Streptococcus suis Infection: From Host Defense to Pathology

Streptococcus suis is a swine pathogen and zoonotic agent responsible for economic losses to the porcine industry. Infected animals may develop meningitis, arthritis, endocarditis, sepsis and/or sudden death. The pathogenesis of the infection implies that bacteria breach mucosal host barriers and reach the bloodstream, where they escape immune-surveillance mechanisms and spread throughout the organism. The clinical manifestations are mainly the consequence of an exacerbated inflammation, defined by an exaggerated production of cytokines and recruitment of immune cells. Among them, neutrophils arrive first in contact with the pathogens to combat the infection. Neutrophils initiate and maintain inflammation, by producing cytokines and deploying their arsenal of antimicrobial mechanisms. Furthermore, neutrophilic leukocytosis characterizes S. suis infection, and lesions of infected subjects contain a large number of neutrophils. Therefore, this cell type may play a role in host defense and/or in the exacerbated inflammation. Nevertheless, a limited number of studies addressed the role or functions of neutrophils in the context of S. suis infection. In this review, we will explore the literature about S. suis and neutrophils, from their interaction at a cellular level, to the roles and behaviors of neutrophils in the infected host in vivo.

Comparative Exoproteome Analysis of Streptococcus suis Human Isolates

The swine pathogen Streptococcus suis is a Gram-positive bacterium which causes infections in pigs, with an impact in animal health and in the livestock industry, and it is also an important zoonotic agent. During the infection process, surface and secreted proteins are essential in the interaction between microorganisms and their hosts. Here, we report a comparative proteomic analysis of the proteins released to the extracellular milieu in six human clinical isolates belonging to the highly prevalent and virulent serotype 2. The total secreted content was precipitated and analyzed by GeLC-MS/MS. In the six strains, 144 proteins assigned to each of the categories of extracellular or surface proteins were identified, as well as 680 predicted cytoplasmic proteins, many of which are putative moonlighting proteins. Of the nine predicted signal peptide-I secreted proteins, seven had relevant antigenic potential when they were analyzed through bioinformatic analysis. This is the first work comparing the exoproteome fraction of several human isolates of this important pathogen.

Integrative gene network analysis identifies TIMP1 as key factors for Haemophilus parasuis infection

Haemophilus parasuis (H.parasuis), an important swine pathogen, causes Glässer's disease leading to pulmonary fibrosis, polyserositis, meningitis, and arthritis. However, the common molecular response and reaction from the host remain unknown. In this study, to uncover novel host factors involved in H.parasuis infection, we identified the global transcriptomics of porcine lung, spleen, blood, alveolar macrophages (PAM), peripheral blood mononuclear cell (PBMC) and aortic vascular endothelial cells (PAVECs) after infection of H.parasuis (Hps0165 strains) using microarray data and high throughput sequencing from Gene Expression Omnibus (GEO), respectively. The results showed that fifteen overlapped genes were significantly regulated in H.parasuis infected porcine lung and spleen, and then were compared with the data from porcine blood, revealing RETN, TIMP1 and C4BPA play potentially an important role for H.parasuis invasion. Furthermore, through analysing porcine cells infected with H.parasuis, we uncover the only overlap gene TIMP1 remarkably upregulated in all assembled data, indicating that TIMP1 could function as key target for the treatment of H.parasuis infection.

A Highly Conserved Epitope (RNNQIPQDF) of Porcine teschovirus Induced a Group-Specific Antiserum: A Bioinformatics-Predicted Model with Pan-PTV Potential

Porcine teschovirus (PTV) is an OIE-listed pathogen with 13 known PTV serotypes. Heterologous PTV serotypes frequently co-circulate and co-infect with another swine pathogen, causing various symptoms in all age groups, thus highlighting the need for a pan-PTV diagnostic tool. Here, a recombinant protein composed of a highly conserved “RNNQIPQDF” epitope on the GH loop of VP1, predicted in silico, and a tandem repeat of this epitope carrying the pan DR (PADRE) and Toxin B epitopes was constructed to serve as a PTV detection tool. This recombinant GST-PADRE-(RNNQIPQDF)n-Toxin B protein was used as an immunogen, which effectively raised non-neutralizing or undetectable neutralizing antibodies against PTV in mice. The raised antiserum was reactive against all the PTV serotypes (PTV–1–7) tested, but not against members of the closely related genera Sapelovirus and Cardiovirus, and the unrelated virus controls. This potential pan-PTV diagnostic reagent may be used to differentiate naturally infected animals from vaccinated animals that have antibodies against a subunit vaccine that does not contain this epitope or to screen for PTV before further subtyping. To our knowledge, this is the first report that utilized in silico PTV epitope prediction to find a reagent broadly reactive to various PTV serotypes.

Comprehensive genome analysis and comparisons of the swine pathogen, Chlamydia suis reveals unique ORFs and candidate host-specificity factors

Chlamydia suis, a ubiquitous swine pathogen, has the potential for zoonotic transmission to humans and often encodes for resistance to the primary treatment antibiotic, tetracycline. Because of this emerging threat, comparative genomics for swine isolate R19 with inter- and intra-species genomes was performed. A 1.094Mb genome was determined through de novo assembly of Illumina high throughput sequencing reads. Annotation and subsystem analyses were conducted, revealing 986 putative genes (Chls_###) that are predominantly orthologs to other known Chlamydia genes. Subsequent comparative genomics revealed a high level of genomic synteny and overall sequence identity with other Chlamydia while 92 unique C. suis open reading frames were annotated. Direct comparison of Chlamydia-specific gene families that included the plasticity zone, inclusion membrane proteins, polymorphic membrane proteins, and the major outer membrane protein, demonstrated high gene content identity with C. trachomatis and C. muridarum. These comparisons also identified diverse components that potentially could contribute to host-specificity. This study constitutes the first genome-wide comparative analysis for C. suis, generating a fully annotated reference genome. These studies will enable focused efforts on factors that provide key species specificity and adaptation to cognate hosts that are attributed to chlamydial infections, including humans.

Update on Streptococcus suis Research and Prevention in the Era of Antimicrobial Restriction: 4th International Workshop on S. suis

Streptococcus suis is a swine pathogen and a zoonotic agent afflicting people in close contact with infected pigs or pork meat. Sporadic cases of human infections have been reported worldwide. In addition, S. suis outbreaks emerged in Asia, making this bacterium a primary health concern in this part of the globe. In pigs, S. suis disease results in decreased performance and increased mortality, which have a significant economic impact on swine production worldwide. Facing the new regulations in preventive use of antimicrobials in livestock and lack of effective vaccines, control of S. suis infections is worrisome. Increasing and sharing of knowledge on this pathogen is of utmost importance. As such, the pathogenesis and epidemiology of the infection, antimicrobial resistance, progress on diagnosis, prevention, and control were among the topics discussed during the 4th International Workshop on Streptococcus suis (held in Montreal, Canada, June 2019). This review gathers together recent findings on this important pathogen from lectures performed by lead researchers from several countries including Australia, Canada, France, Germany, Japan, Spain, Thailand, The Netherlands, UK, and USA. Finally, policies and recommendations for the manufacture, quality control, and use of inactivated autogenous vaccines are addressed to advance this important field in veterinary medicine.

Examining the Effect of Host Recruitment Rates on the Transmission of Streptococcus suis in Nursery Swine Populations

Streptococcus suis is a swine pathogen that is capable of causing severe outbreaks of disease in the nursery. Demographic parameters such as host recruitment rates can have profound effects on the transmission dynamics of infectious diseases and, thus, are critically important in high-turnover populations such as farmed swine. However, knowledge concerning the implications that such parameters have on S. suis disease control remains unknown. A stochastic mathematical model incorporating sub-clinically infected pigs was developed to capture the effects of changes in host recruitment rate on disease incidence. Compared to our base model scenario, our results show that monthly introduction of pigs into the nursery (instead of weekly introduction) reduced cumulative cases of S. suis by up to 59%, while increasing disease-removal rates alone averted up to 64% of cases. Sensitivity analysis demonstrated that the course of infection in sub-clinically infected pigs was highly influential and generated significant variability in the model outcomes. Our model findings suggest that modifications to host recruitment rates could be leveraged as a tool for S. suis disease control, however improving our understanding of additional factors that influence the risk of transmission would improve the precision of the model estimates.