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State-of-the-Art Animal Virus Research in Poland II

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A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Animal Viruses".

Deadline for manuscript submissions: closed (31 May 2023) | Viewed by 21939

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Prof. Dr. Jacek Kuźmak

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National Veterinary Reserach Institute, Pulawy, Poland
Interests: ruminant retroviruses; epidemiology; diagnosis; host–virus interaction
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Dr. Magdalena Materniak-Kornas

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Department of Biochemistry, National Veterinary Research Institute, Al. Partyzantów 57, 24-100 Puławy, Poland
Interests: molecular diagnostics; serological diagnostics; ruminant retroviruses; transcriptomics
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Dear Colleagues,

Poland, a large country in the middle of Europe, has a very interesting history and a variety of landscapes and ecosystems. Thanks to these features, Poland has intensive animal production and trade, and this industrial focus has been accompanied by extensive research in animal virology. The eradication of several animal diseases in the last decades is a great success, but as we have experienced recently, nature is unpredictable. Poland, like other countries, struggles with several emerging and re-emerging viruses in the field of animal health. Currently, new technologies and concepts allow multifactorial analyses of pathogen biology and interaction with the host, providing novel insights into pathogenesis and causality which may lead to new prevention or treatment strategies.

Hereby we announce the second edition of the Special Issue entitled “State-of-the-Art Animal Virus Research in Poland II”, dedicated to the recent research in the field of veterinary virology in Poland. We cordially invite Polish scientists to contribute their original studies, communications, case studies, and review articles.

Prof. Dr. Jacek Kuźmak
Dr. Magdalena Materniak-Kornas
Guest Editors

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Keywords

epidemiology
pathogenesis
diagnostics
(immuno)pathology
molecular and omics aspects
virus evolution
virus-host interactions
prophylaxis and prevention
zoonotic potential

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Published Papers (9 papers)

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15 pages, 842 KiB

Open AccessArticle

Digital PCR (dPCR) Quantification of miR-155-5p as a Potential Candidate for a Tissue Biomarker of Inflammation in Rabbits Infected with Lagovirus europaeus/Rabbit Hemorrhagic Disease Virus (RHDV)

by Beata Hukowska-Szematowicz, Ewa Ostrycharz, Wioleta Dudzińska, Paulina Roszkowska, Aldona Siennicka and Iwona Wojciechowska-Koszko

Viruses 2023, 15(7), 1578; https://doi.org/10.3390/v15071578 - 19 Jul 2023

Cited by 6 | Viewed by 2425

Abstract

MicroRNAs (miRNAs, miRs) are a group of small, 17–25 nucleotide, non-coding RNA sequences that, in their mature form, regulate gene expression at the post-transcriptional level. They participate in many physiological and pathological processes in both humans and animals. One such process is viral infection, in which miR-155 participates in innate and adaptive immune responses to a broad range of inflammatory mediators. Recently, the study of microRNA has become an interesting field of research as a potential candidate for biomarkers for various processes and disease. To use miRNAs as potential biomarkers of inflammation in viral diseases of animals and humans, it is necessary to improve their detection and quantification. In a previous study, using reverse transcription real-time quantitative PCR (RT-qPCR), we showed that the expression of ocu-miR-155-5p in liver tissue was significantly higher in rabbits infected with Lagovirus europaeus/Rabbit Hemorrhagic Disease Virus (RHDV) compared to healthy rabbits. The results indicated a role for ocu-miR-155-5p in Lagovirus europaeus/RHDV infection and reflected hepatitis and the impairment/dysfunction of this organ during RHD. MiR-155-5p was, therefore, hypothesized as a potential candidate for a tissue biomarker of inflammation and examined in tissues in Lagovirus europaeus/RHDV infection by dPCR. The objective of the study is the absolute quantification of ocu-miR-155-5p in four tissues (liver, lung, kidney, and spleen) of rabbits infected with Lagovirus europaeus/RHDV by digital PCR, a robust technique for the precise and direct quantification of small amounts of nucleic acids, including miRNAs, without standard curves and external references. The average copy number/µL (copies/µL) of ocu-miRNA-155-5p in rabbits infected with Lagovirus europaeus GI.1a/Rossi in the liver tissue was 12.26 ± 0.14, that in the lung tissue was 48.90 ± 9.23, that in the kidney tissue was 16.92 ± 2.89, and that in the spleen was 25.10 ± 0.90. In contrast, in the tissues of healthy control rabbits, the average number of copies/µL of ocu-miRNA-155-5p was 5.07 ± 1.10 for the liver, 23.52 ± 2.77 for lungs, 8.10 ± 0.86 for kidneys, and 42.12 ± 3.68 for the spleen. The increased expression of ocu-miRNA-155-5p in infected rabbits was demonstrated in the liver (a fold-change of 2.4, p-value = 0.0003), lung (a fold-change of 2.1, p-value = 0.03), and kidneys (a fold-change of 2.1, p-value = 0.01), with a decrease in the spleen (a fold-change of 0.6, p-value = 0.002). In the study of Lagovirus europaeus/RHDV infection and in the context of viral infections, this is the first report that shows the potential use of dPCR for the sensitive and absolute quantification of microRNA-155-5p in tissues during viral infection. We think miR-155-5p may be a potential candidate for a tissue biomarker of inflammation with Lagovirus europaeus/RHDV infection. Our report presents a new path in discovering potential candidates for the tissue biomarkers of inflammation. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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13 pages, 1954 KiB

Open AccessArticle

Phylogeny of Shrew- and Mole-Borne Hantaviruses in Poland and Ukraine

by Fuka Kikuchi, Satoru Arai, Janusz Hejduk, Ai Hayashi, Janusz Markowski, Marcin Markowski, Leszek Rychlik, Vasyl Khodzinskyi, Hajime Kamiya, Tetsuya Mizutani, Motoi Suzuki, Beata Sikorska, Paweł P. Liberski and Richard Yanagihara

Viruses 2023, 15(4), 881; https://doi.org/10.3390/v15040881 - 29 Mar 2023

Cited by 1 | Viewed by 1899

Abstract

Earlier, we demonstrated the co-circulation of genetically distinct non-rodent-borne hantaviruses, including Boginia virus (BOGV) in the Eurasian water shrew (Neomys fodiens), Seewis virus (SWSV) in the Eurasian common shrew (Sorex araneus) and Nova virus (NVAV) in the European mole (Talpa europaea), in central Poland. To further investigate the phylogeny of hantaviruses harbored by soricid and talpid reservoir hosts, we analyzed RNAlater^®-preserved lung tissues from 320 shrews and 26 moles, both captured during 1990–2017 across Poland, and 10 European moles from Ukraine for hantavirus RNA through RT-PCR and DNA sequencing. SWSV and Altai virus (ALTV) were detected in Sorex araneus and Sorex minutus in Boginia and the Białowieża Forest, respectively, and NVAV was detected in Talpa europaea in Huta Dłutowska, Poland, and in Lviv, Ukraine. Phylogenetic analyses using maximum-likelihood and Bayesian methods showed geography-specific lineages of SWSV in Poland and elsewhere in Eurasia and of NVAV in Poland and Ukraine. The ATLV strain in Sorex minutus from the Białowieża Forest on the Polish–Belarusian border was distantly related to the ATLV strain previously reported in Sorex minutus from Chmiel in southeastern Poland. Overall, the gene phylogenies found support long-standing host-specific adaptation. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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15 pages, 4066 KiB

Open AccessArticle

Epidemiology of Infectious Bursal Disease Virus in Poland during 2016–2022

by Anna Pikuła, Anna Lisowska and Katarzyna Domańska-Blicharz

Viruses 2023, 15(2), 289; https://doi.org/10.3390/v15020289 - 19 Jan 2023

Cited by 3 | Viewed by 2522

Abstract

Infectious bursal disease virus is an immunosuppressive ubiquitous pathogen that causes serious economic losses in poultry production. The virus is prone to genetic changes through mutations and reassortment, which drive the emergence of new variants and lead to a change in the epidemiological situation in a field. Such a situation is currently being reported due to a large wave of IBDV A3B1 reassortant infections in northwestern Europe. On the other hand, in Poland, which is the largest producer of chicken meat in the EU, the IBDVs of genotypes A3B2 and A3B4 were circulating just before the emergence of A3B1 reassortants. The purpose of the presented study was to update the IBDV epidemiological situation. The performed molecular survey based on the sequence of both genome segments showed the presence of very virulent strains (A3B2) and reassortants of genotypes A3B4 and A3B1; moreover, two of these genotypes are newly introduced IBDV lineages. In addition, a number of amino acid substitutions were demonstrated, including within antigenic epitopes and virulence determinants. In conclusion, the results obtained indicated a dynamic epidemiological situation in Poland, which highlights the need for further monitoring studies in the region and verification of protection conferred by the vaccines used against infection with detected IBDV. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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13 pages, 1066 KiB

Open AccessArticle

European Brown Hare Syndrome in Poland: Current Epidemiological Situation

by Andrzej Fitzner, Wiesław Niedbalski, Andrzej Kęsy, Bogusław Rataj and Marian Flis

Viruses 2022, 14(11), 2423; https://doi.org/10.3390/v14112423 - 31 Oct 2022

Cited by 5 | Viewed by 1885

Abstract

European brown hare syndrome (EBHS) is one of the main causes of mortality in brown hares (Lepus europaeus) and mountain hares (Lepus timidus) in Europe. Since the mid-1990s, this highly lethal and contagious plague has been widespread in many European countries, contributing to a drastic decline in the number of free-living and farmed hares. A second lagovirus, able to infect some species of hares is rabbit haemorrhagic disease virus 2 (RHDV2; GI.2) recognised in 2010, a new viral emergence of RHDV (GI.1) which is known to be responsible for haemorrhagic disease in rabbits—RHD. The aim of this study was to evaluate the current EBHS epidemiological situation on the basis of the presence of antibodies to European brown hare syndrome virus (EBHSV) and anti-RHDV2 antibodies in sera collected from free-ranging hares in Central and Southeastern Poland in 2020–2021. Additionally, studies on the presence of EBHSV and RHDV2 antigens or their genetic material in the blood and internal organs taken from brown hares between 2014–2021 have been carried out. The results of the serological examination showed nearly 88% of tested blood samples were positive for EBHSV antibodies. No EBHSV was identified in the examined hares using virological and molecular tests. The positive results of EBHS serological studies confirmed the circulation and maintenance of EBHSV in free-living brown hares in Poland. However, no serological, virological or molecular evidence was obtained indicating that the brown hares tested had been in contact with RHDV2. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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18 pages, 1357 KiB

Open AccessArticle

Occurrence of Equine Foamy Virus Infection in Horses from Poland

by Magdalena Materniak-Kornas, Wojciech Rożek, Jerzy Rola, Zbigniew Osiński, Martin Löchelt and Jacek Kuźmak

Viruses 2022, 14(9), 1973; https://doi.org/10.3390/v14091973 - 6 Sep 2022

Viewed by 1698

Abstract

Equine foamy virus (EFVeca) is a foamy virus of non-primate origin and among the least-studied members of this retroviral subfamily. By sequence comparison, EFVeca shows the highest similarity to bovine foamy virus. In contrast to simian, bovine or feline foamy viruses, knowledge about the epidemiology of EFVeca is still limited. Since preliminary studies suggested EFVeca infections among horses in Poland, we aimed to expand the diagnostics of EFVeca infections by developing specific diagnostic tools and apply them to investigate its prevalence. An ELISA test based on recombinant EFVeca Gag protein was developed for serological investigation, while semi-nested PCR for the detection of EFVeca DNA was established. 248 DNA and serum samples from purebred horses, livestock and saddle horses, Hucul horses and semi-feral Polish primitive horses were analyzed in this study. ELISA was standardized, and cut off value, sensitivity and specificity of the test were calculated using Receiver Operating Characteristic and Bayesian estimation. Based on the calculated cut off, 135 horses were seropositive to EFVeca Gag protein, while EFVeca proviral DNA was detected in 85 animals. The rate of infected individuals varied among the horse groups studied; this is the first report confirming the existence of EFVeca infections in horses from Poland using virus-specific tools. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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20 pages, 3452 KiB

Open AccessArticle

Transcriptomic Analysis of MDBK Cells Infected with Cytopathic and Non-Cytopathic Strains of Bovine Viral Diarrhea Virus (BVDV)

by Paweł Mirosław, Marzena Rola-Łuszczak, Jacek Kuźmak and Mirosław P. Polak

Viruses 2022, 14(6), 1276; https://doi.org/10.3390/v14061276 - 11 Jun 2022

Cited by 8 | Viewed by 3038

Abstract

Bovine viral diarrhea virus (BVDV) belongs to the Flaviviridae family and the Pestivirus genus. Infection with BVDV causes a disease with a wide spectrum of clinical symptoms, most often mild, although infections with this virus constitute a serious economic problem all over the world. The virus is characterized by a high genetic variability, while the accumulation of single mutations leads to the formation of its new variants. The aim of this study was to better understand the complicated pathogenesis of this disease at the molecular level via the analysis of the transcriptome of cells infected with this virus. The bovine kidney cell line (MDBK), the cytopathic (cp) reference strain, and two non-cytopathic (ncp) BVD virus field strains were used in transcriptomic studies. The cell transcriptome was tested 24 and 72 h after infection. The results of the microarray analysis revealed changes in the expression levels of numerous genes. Genes with changed expression as a result of infection with the cp strain caused changes in the expression levels of a large number of genes and enriched a number of pathways. Genes with increased expression levels were enriched among other pathways involved in the cell cycle, while genes with reduced expression levels enriched pathways mostly related to metabolism. Genes with increased expression levels as a result of infection with ncp strains enriched a much smaller number of pathways, among them, pathways related to signaling activity 24 h post-infection and serine biosynthetic pathways both 24 and 72 h post-infection. Pathways enriched by genes with reduced expression levels were related to the innate immune response (72 h post-infection) or metabolism (24 and 72 h post-infection). The results of microarray studies can help us to better understand the host’s response to BVDV infection. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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16 pages, 2144 KiB

Open AccessArticle

Functional Analysis of a Frontal miRNA Cluster Located in the Large Latency Transcript of Pseudorabies Virus

by Weronika Hoffmann, Andrea D. Lipińska and Krystyna Bieńkowska-Szewczyk

Viruses 2022, 14(6), 1147; https://doi.org/10.3390/v14061147 - 26 May 2022

Cited by 1 | Viewed by 2206

Abstract

MicroRNAs (miRNAs) have been identified as a class of crucial regulators of virus-host crosstalk, modulating such processes as viral replication, antiviral immune response, viral latency, and pathogenesis. Pseudorabies virus (PRV), a model for the study of alphaherpesvirus biology, codes for 11 distinct miRNAs mapped to the ~4.6 kb intron of Large Latency Transcript (LLT). Recent studies have revealed the role of clusters consisting of nine and eleven miRNA genes in the replication and virulence of PRV. The function of separate miRNA species in regulating PRV biology has not been thoroughly investigated. To analyze the regulatory potential of three PRV miRNAs located in the frontal cluster of the LLT intron, we generated a research model based on the constitutive expression of viral miRNAs in swine testis cells (ST_LLT [1–3] cell line). Using a cell culture system providing a stable production of individual miRNAs at high levels, we demonstrated that the LLT [1–3] miRNA cluster significantly downregulated IE180, EP0, and gE at the early stages of PRV infection. It was further determined that LLT [1–3] miRNAs could regulate the infection process, leading to a slight distortion in transmission and proliferation ability. Collectively, our findings indicate the potential of LLT [1–3] miRNAs to retard the host responses by reducing viral antigenic load and suppressing the expansion of progeny viruses at the early stages of infection. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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20 pages, 4442 KiB

Open AccessArticle

Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

by Beata Gromadzka, Milena Chraniuk, Lilit Hovhannisyan, Karolina Uranowska, Bogusław Szewczyk, Magdalena Narajczyk and Mirosława Panasiuk

Viruses 2022, 14(4), 730; https://doi.org/10.3390/v14040730 - 30 Mar 2022

Cited by 2 | Viewed by 3333

Abstract

(1) Background: Avian influenza viruses (AIVs) promptly evade preexisting immunity by constantly altering the immunodominant neutralizing antibody epitopes (antigenic drift) or by procuring new envelope serotypes (antigenic shift). As a consequence, the majority of antibodies elicited by infection or vaccination protect only against closely related strains. The immunodominance of the globular head of the main glycoprotein has been shown to mask the immunogenicity of the conserved regions located within the hemagglutinin (HA) protein. It has been shown that the broadly neutralizing universal antibodies recognize the HA2 domain in headless hemagglutinin (HA-stalk). Therefore, the HA-stalk is a highly conserved antigen, which makes it a good candidate to be used in universal vaccine development against AIVs. (2) Methods: Sf9 insect cells were used to produce triple H5N1/NA-HA-M1 influenza virus-like particles (VLPs) via co-expression of neuraminidase, hemagglutinin and matrix proteins from a tricistronic expression cassette. Purified influenza VLPs were used to immunize broiler hens. An in-depth characterization of the immune response was performed with an emphasis on the pool of elicited universal antibodies. (3) Results: Our findings suggest, that after vaccination with triple H5N1/NA-HA-M1 VLPs, hens generate a pool of broad-spectrum universal anti-HA-stalk antibodies. Furthermore, these universal antibodies are able to recognize the mammalian-derived HA-stalk recombinant proteins from homologous H5N1 and heterologous H7N9 AIVs as well as from the heterosubtypic human H1N1 influenza strain. (4) Conclusions: Our findings may suggest that highly pathogenic avian influenza H5 HA protein contain functional epitopes that are attractive targets for the generation of broad-spectrum antibodies against AIVs in their native hosts. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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13 pages, 2847 KiB

Open AccessBrief Report

Genetic Diversity of the LTR Region of Polish SRLVs and Its Impact on the Transcriptional Activity of Viral Promoters

by Monika Olech and Jacek Kuźmak

Viruses 2023, 15(2), 302; https://doi.org/10.3390/v15020302 - 22 Jan 2023

Cited by 2 | Viewed by 1751

Abstract

A long terminal repeat (LTR) plays an indispensable role in small ruminant lentivirus (SRLV) gene expression. In this study, we present the LTR sequence of Polish SRLVs representing different subtypes, and analyzed their impact on SRLV promoter activity, as measured in transient transfection assays. Although certain nucleotide motifs (AML(vis), TATA box and the polyadenylation site (AATAAA)) were conserved across sequences, numerous mutations within the LTR sequences have been identified. Single nucleotide polymorphisms (SNPs) were detected in both regulatory (AP-1, AP-4, Stat and Gas) and non-regulatory sequences, and subtype-specific genetic diversity in the LTR region of Polish SRLVs was observed. In vitro assays demonstrated subtype-specific functional differences between the LTR regions of distinct SRLV subtypes. Our results revealed that the promoter activity of Polish strains was lower (1.64–10.8-fold) than that noted for the K1514 reference strain; however, the differences in most cases were not statistically significant. The lowest promoter activity was observed for strains representing subtype A5 (mean 69.067) while the highest promoter activity was observed for strain K1514 representing subtype A1 (mean 373.48). The mean LTR activities of strains representing subtypes A12, A17, A23, A18 and A24 were 91.22, 137.21, 178.41, 187.05 and 236.836, respectively. The results of the inter-subtype difference analysis showed that the promoter activity of strains belonging to subtype A5 was significantly lower than that for subtype A12 strains (1.32-fold; p < 0.00). The promoter activities of the A5 strain were 1.98-fold and 2.58-fold less active than that of the A17 and A23 strains, and the promoter activities of A12 strains were 1.955 and 1.5 times lower than the promoter activity of A23 and A17 strains, respectively. Furthermore, the promoter activity of A17 strains was 1.3 lower than the promoter activity of A23 strains. Our findings suggest that subtype-specific genetic diversity, mainly in the transcription factor’s binding sites, has an impact on their transcriptional activity, producing a distinct activity pattern for the subtypes. This study provides new information that is important for better understanding the function of the SRLV LTR. However, further research including more strains and subtypes as well as other cell lines is needed to confirm these findings. Full article

(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland II)

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