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Keywords = small ruminant lentivirus (SRLV)

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12 pages, 1063 KiB  
Article
Identification of New Single Nucleotide Polymorphisms Potentially Related to Small Ruminant Lentivirus Infection Susceptibility in Goats Based on Data Selected from High-Throughput Sequencing
by Magdalena Materniak-Kornas, Katarzyna Ropka-Molik, Katarzyna Piórkowska, Joanna Kowalik, Tomasz Szmatoła, Jacek Sikora, Aldona Kawęcka and Jacek Kuźmak
Pathogens 2024, 13(10), 830; https://doi.org/10.3390/pathogens13100830 - 25 Sep 2024
Viewed by 188
Abstract
Small ruminant lentivirus (SRLV) infections are spread in the flocks of sheep and goats all over the world, causing economic loss. Although only a fraction of infected animals develop disease symptoms, all of them may shed the virus, causing uncontrolled spread of the [...] Read more.
Small ruminant lentivirus (SRLV) infections are spread in the flocks of sheep and goats all over the world, causing economic loss. Although only a fraction of infected animals develop disease symptoms, all of them may shed the virus, causing uncontrolled spread of the infection. Antibodies against the virus can be detected in the blood of infected animals and are the main marker of infection. Additionally, in most infected animals, proviral DNA can also be detected, but at different levels. Due to the lack of treatment or vaccines, the most effective strategy to prevent SRLV infections are control programmes introduced by several countries based on the elimination of seropositive individuals from the flock. An alternative approach, which has currently become the rationale, is an identification of host factors which may predispose certain individuals or breeds to resistance or susceptibility to small ruminant lentivirus infection. In our work, attention was paid to goats of the Carpathian breed infected with SRLV. Available RNA-seq results from the blood of 12 goats with a determined level of SRLV proviral load were used to analyse single nucleotide polymorphisms (SNPs) by the variant calling method. Six SNPs within five genes (POU2AF1, BCAT2, TMEM154, PARP14, UBASH3A) were selected for genotyping to determine their association with the level of small ruminant lentivirus proviral DNA in a group of 60 goats. Interestingly, in seronegative individuals, only the TT genotype of the PARP14 gene was observed, while the TMEM154 CC genotype was found only in seropositive goats. Both genes may be considered potential markers for resistance/susceptibility to SRLV infection. In contrast, polymorphisms identified in POU2AF1 and UBASH3A genes seemed to be deleterious for respective protein functions; therefore, these genes are less likely to be recognised as resistance/susceptibility markers of SRLV infection. Full article
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<p>Density plot depicting the distribution of identified genetic variants.</p>
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<p>The function and location of identified polymorphisms with their percentage share.</p>
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<p>The genotype distribution for SNPs detected in UBASH3A, POU2AF1, BCAT2, TMEM154 and PARP14 genes.</p>
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13 pages, 900 KiB  
Article
Cross-Sectional Study Assessing Management Practices and Udder Health in California Sheep Flocks and Seroprevalence of Small Ruminant Lentivirus
by Rose Digianantonio, Alda F. A. Pires and Roselle Busch
Animals 2024, 14(16), 2332; https://doi.org/10.3390/ani14162332 - 13 Aug 2024
Viewed by 633
Abstract
(1) Background: Information is lacking on small ruminant lentivirus (SRLV) status, prevalence, risk factors, and control measures for mastitis in California ewes. The goal of this survey was to outline characteristics of the sheep industry in California related to udder health and mastitis [...] Read more.
(1) Background: Information is lacking on small ruminant lentivirus (SRLV) status, prevalence, risk factors, and control measures for mastitis in California ewes. The goal of this survey was to outline characteristics of the sheep industry in California related to udder health and mastitis management. (2) Methods: An online survey consisting of 48 questions was completed by respondents between April 2022 and February 2023. Descriptive analysis and chi-squared tests were conducted to evaluate associations between variables. A multiple correspondence analysis (MCA) of general management practices, udder health management, and flock demographics was performed to assess clustering. A subset of respondents (20) participated in SRLV serology testing. (3) Results: Seventy-one completed surveys were submitted. The MCA showed two clusters. Larger flock sizes, the use of breeding ewes for meat or wool production or contract grazing, and extensive management practices were more closely related to >5% udder abnormalities per lactation and ≥5% orphan lambs. The flock-level seroprevalence of SRLV was 75% (15/20), and ewe-level seroprevalence was 14.1% (183/1106). (4) Conclusions: The results of this study highlight areas that need further research, such as exploring differences in mastitis and SRLV incidences among management systems, the efficacy of mastitis treatments, and education on critical timepoints for mastitis diagnosis and control. Full article
(This article belongs to the Collection Diseases of Small Ruminants)
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<p>Distributions of survey responses by region in California and flock size.</p>
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<p>Multiple correspondence analysis of 10 flock-level variables. Quadrants are labeled by number. The color scale represents the contribution of the response to the variance in the dataset. Yellow represents low contribution and navy represents high contribution to variance.</p>
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21 pages, 2513 KiB  
Article
Development of a Bead-Based Multiplex Fluorescent Immunoassay to Detect Antibodies against Maedi-Visna Virus in Sheep
by Anniken Jerre Borge, Barbara Colitti, Sergio Rosati, Anne B. Nordstoga, Britt Gjerset, Kristin Udjus, Chiara Nogarol, Stalin Chellappa, Ingunn Anita Samdal and Kari Lybeck
Animals 2024, 14(10), 1442; https://doi.org/10.3390/ani14101442 - 12 May 2024
Viewed by 1043
Abstract
The Maedi-visna virus (MVV) causes a persistent infection in small ruminants, and its high genetic heterogeneity affects the performance of diagnostic tests when used in different populations. Therefore, the aim of this study was to develop a bead-based multiplex immunoassay tailored to detect [...] Read more.
The Maedi-visna virus (MVV) causes a persistent infection in small ruminants, and its high genetic heterogeneity affects the performance of diagnostic tests when used in different populations. Therefore, the aim of this study was to develop a bead-based multiplex immunoassay tailored to detect antibodies against a Norwegian MVV strain. We used tissue samples from 14 PCR-positive sheep from a recent MVV outbreak in Norway to sequence the viral strain and produced recombinant antigens based on sequences from one animal. The assay included commercial TM-A and recombinant Norwegian p25, p16–25 and SU5 antigens. Cut-off values for each antigen were determined using receiver operating characteristic curves on 40 ELISA-negative and 67 ELISA-positive samples from the outbreak. The intraplate and interplate repeatability were investigated by testing a quadruplicate of five samples over three days, while the analytical sensitivity (aSe) and specificity (aSp) were measured in comparison to a commercial ELISA. The repeatability showed a coefficient of variation below 15% for most positive samples. The aSe was equal or higher for the multiplex assay than the ELISA, and the aSp of each antigen was 91.7, 93.3, 95.0 and 93.3% for p25, p16–25, SU5 and TM-A, respectively. The assay shows promising results; however, further evaluations of diagnostic characteristics are necessary before implementation in the Norwegian surveillance programme. Full article
(This article belongs to the Special Issue Small Ruminants and Lentivirus Research: Future Directions)
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Graphical abstract
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<p>Coomassie blue staining of SDS-PAGE gel showing the Norwegian recombinant proteins with the expected molecular sizes of 29 kDa, 66 kDa, and 50 kDa for SU5 (lane 1), p16–25 (lane 2) and p25 (lane 3), respectively. The ladder is represented in lane 4.</p>
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<p>The median fluorescence intensity (MFI) of ELISA-positive samples (grey) and ELISA-negative samples (black) for all antigens and controls (GST and the BSA-coated control beads) in the bead-based immunoassay when MFI signal from the blank wells were subtracted. The samples were from panel two and were diluted 1:100. The amount of antigen coupled to beads was 8 µg for the TM-A antigen, the commercial p16–25, the Norwegian p16–25, and the Norwegian SU5 antigen, while 12 µg was coupled to the commercial p25, the Norwegian p25 and the GST-coated bead.</p>
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<p>Corrected median fluorescence intensity (corrected MFI) in the bead-based immunoassay for the beads with the Norwegian recombinant antigens p25, p16–25 and SU5, and the commercial TM-A antigen, when tested with twofold dilutions (1:25 to 1:800) of samples from panel 3. Corrected MFI was the MFI signal after subtraction of the MFI signal from the blank wells and the MFI signal of the BSA-coated control bead. The grey dots represent ELISA-positive samples (10 samples), and the black dots represent ELISA-negative samples (8 samples).</p>
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<p>The distribution of S/P% for the Norwegian recombinant p25, p16–25 and SU5, and the commercial and TM-A antigen in the bead-based multiplex immunoassay when testing samples from panel 5. The black bars represent ELISA-negative samples (<b>left panel</b>), while the grey bars represent ELISA-positive samples (<b>right panel</b>). The S/P% values are shown on the <span class="html-italic">x</span>-axis, and the <span class="html-italic">y</span>-axis illustrates the number of samples within each S/P% value category (bin width 100). The cut-off values for each antigen are illustrated as red vertical lines.</p>
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<p>Distribution of S/P% for each antigen p25, p16–25, SU5 and TM-A in the bead-based multiplex immunoassay after analysis of 60 ELISA-negative samples (black dots). The red dots represent the same sample that was positive for all three antigens. The black horizontal lines represent the cut-off values for each antigen.</p>
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<p>ELISA-positive samples from six flocks diagnosed with MVV in the 2019–2020 outbreak with a seroprevalence above 30% (15 samples from flocks A, C, E and F, 19 samples from flock G and 18 samples from flock I, from panels 5 and 8) were analysed by the bead-based multiplex immunoassay including antigens p25, p16–25, SU5 and TM-A. The flocks are defined as A–I, which corresponds to flock A–I in [<a href="#B25-animals-14-01442" class="html-bibr">25</a>]. The horizontal lines represent the cut-off values for each antigen.</p>
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<p>The S/P% from the bead-based multiplex immunoassay with antigens p25, p16–25, SU5 and TM-A, following the testing of samples positive in the screening and Elitest ELISA but negative in the verification ELISA (panel 9). The samples originated from flocks diagnosed with MVV (grey dots, 15 samples from 6 different positive flocks) and flocks without an MVV diagnosis (black dots, 14 samples from 12 different negative flocks). The horizontal lines represent the cut-off value for each antigen.</p>
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13 pages, 2847 KiB  
Brief 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 1657
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 [...] Read more.
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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<p>Neighbor-joining phylogenetic tree based on the alignment of the LTR fragment. Sequences from this study are labeled with black circles (s—sheep; g—goat). Reference SRLV strains are shown by name followed by their GenBank accession number and subtype. Numbers at the branches indicate the percentage of bootstrap values obtained from 1000 replicates.</p>
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<p>Alignment of U3-R sequences of the LTR region from Polish SRLV strains analyzed in this study. Dots indicate an identical nucleotide, and dashes represent gaps. Boundaries between U3, R and U5 are indicated by straight arrows. AP-1, AP-4, AML (vis) motifs, TATA box, GAS, TAS, Stat-1 and polyadenylation signal (poly A) are marked by boxes. NF-κB, IFR-1 and CAAT are shaded and indicated by a solid bar. s—sheep; g—goat.</p>
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<p>Alignment of U3-R sequences of the LTR region from Polish SRLV strains analyzed in this study. Dots indicate an identical nucleotide, and dashes represent gaps. Boundaries between U3, R and U5 are indicated by straight arrows. AP-1, AP-4, AML (vis) motifs, TATA box, GAS, TAS, Stat-1 and polyadenylation signal (poly A) are marked by boxes. NF-κB, IFR-1 and CAAT are shaded and indicated by a solid bar. s—sheep; g—goat.</p>
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<p>Alignment of U3-R sequences of the LTR region from Polish SRLV strains analyzed in this study. Dots indicate an identical nucleotide, and dashes represent gaps. Boundaries between U3, R and U5 are indicated by straight arrows. AP-1, AP-4, AML (vis) motifs, TATA box, GAS, TAS, Stat-1 and polyadenylation signal (poly A) are marked by boxes. NF-κB, IFR-1 and CAAT are shaded and indicated by a solid bar. s—sheep; g—goat.</p>
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<p>Basal transcriptional activity of SRLV LTR variants in TIGEF cells. Results present the mean of three independent transfection experiments, and standard deviations are shown as error bars. The asterisk indicates a significant difference (<span class="html-italic">p</span> &lt; 0.05) with the reference strain K1514 LTR according to the Mann–Whitney U Test.</p>
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<p>Subtype-specific transcriptional activity of Polish SRLV LTR variants in TIGEF cells. Differential activity of the SRLV LTR variants representing subtypes A1, A5, A12, A17, A18, A23 and A24. Results present the mean of independent transfection experiments, and standard deviations are shown as error bars. The asterisk indicates a significant difference (<span class="html-italic">p</span> &lt; 0.05) according the Mann–Whitney U test. Subtypes A1, A17, A18, A23 and A24 were not included in the inter-subtype analysis due to limited samples (&lt;5) representing each subtype.</p>
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9 pages, 373 KiB  
Article
Loss of Caprine Arthritis Encephalitis Virus (CAEV) Herd Accreditation: Characteristics, Diagnostic Approach, and Specific Follow-Up Scenarios on Large Dairy Goat Farms
by Karianne Peterson, René van den Brom, Marian Aalberts, Carlijn ter Bogt-Kappert and Piet Vellema
Pathogens 2022, 11(12), 1541; https://doi.org/10.3390/pathogens11121541 - 15 Dec 2022
Cited by 3 | Viewed by 1700
Abstract
The retrovirus causing caprine arthritis encephalitis (CAE), a slowly progressive inflammatory disease in goats, belongs to the group of small ruminant lentiviruses (SRLVs) which cause lifelong infections that ought to be avoided for animal welfare as well as economic reasons. SRLV accreditation has [...] Read more.
The retrovirus causing caprine arthritis encephalitis (CAE), a slowly progressive inflammatory disease in goats, belongs to the group of small ruminant lentiviruses (SRLVs) which cause lifelong infections that ought to be avoided for animal welfare as well as economic reasons. SRLV accreditation has been in place for forty years in The Netherlands and is based on the screening of small ruminant sera for specific antibodies. This paper evaluates 38 dairy goat herds that lost CAEV accreditation between 2012 and 2022. The characteristics of these herds are discussed, and specific follow-up scenarios, depending on desired goals, are introduced. The herd size of the participating herds varies from approximately 400 to 4600 adult dairy goats. The larger herds tended to be more prone to lose herd accreditation and had more difficulties regaining accreditation. Possible routes of introduction are lined up. The Royal GD’s tailor-made approach and advice to support livestock farmers with herds that have lost CAE accreditation are discussed in detail. Specific emphasis is placed on the strategic deployment of various diagnostic tests (such as antibody ELISAs and PCR) in different media, such as (pooled) sera, (bulk)milk and tissue samples. Special attention is paid to the added value of retrospective bulk milk testing or the specific testing of groups based on housing and management, which enables the investigation of the moment of viral introduction and route of transmission into a herd. Furthermore, the prospective implementation of bulk milk and strategic pooled milk sample testing in the Dutch SRLV accreditation programs intensifies surveillance and enables the taking of swift action to prevent further transmission within and between herds. An appeal is made to share experiences to improve programs collectively, and to start research into the underlying mechanisms. Full article
(This article belongs to the Section Viral Pathogens)
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<p>Bulk milk testing for caprine arthritis encephalitis virus antibodies (ELISA) in herds that lost CAEV accreditation.</p>
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16 pages, 2938 KiB  
Article
The Prevalence of Histopathological Features of Pneumonia in Goats with Symptomatic Caprine Arthritis-Encephalitis
by Agata Moroz, Michał Czopowicz, Małgorzata Sobczak-Filipiak, Izabella Dolka, Magdalena Rzewuska, Magdalena Kizerwetter-Świda, Dorota Chrobak-Chmiel, Marcin Mickiewicz, Lucjan Witkowski, Olga Szaluś-Jordanow, Tomasz Nalbert, Adrian Valentin Potârniche, Karolina Barszcz, Iwona Markowska-Daniel, Ryszard Puchała, Emilia Bagnicka and Jarosław Kaba
Pathogens 2022, 11(6), 629; https://doi.org/10.3390/pathogens11060629 - 30 May 2022
Cited by 6 | Viewed by 2984
Abstract
Chronic interstitial pneumonia (CIP) is a main pathology of sheep infected with small ruminant lentivirus (SRLV). Caprine arthritis-encephalitis (CAE) is caused by the same pathogen; however, the presence of CIP has been only occasionally reported in SRLV-infected goats. We carried out a cross-sectional [...] Read more.
Chronic interstitial pneumonia (CIP) is a main pathology of sheep infected with small ruminant lentivirus (SRLV). Caprine arthritis-encephalitis (CAE) is caused by the same pathogen; however, the presence of CIP has been only occasionally reported in SRLV-infected goats. We carried out a cross-sectional study to determine the prevalence of histopathological lesions indicative of CIP in goats with symptomatic CAE, and to investigate whether CIP was associated with a higher prevalence of other types of pneumonia (purulent bronchopneumonia, fibrinous pleuropneumonia) or bacterial infections. Lung specimens and bronchial swabs were collected for histopathological and bacteriological examination, respectively, from 116 goats from a CAE-affected herd. All goats were euthanized due to severe clinical signs of CAE. The goats were seropositive for SRLV infection in two different ELISAs and the presence of SRLV antigen in the lung tissue was confirmed by immunohistochemistry. Histopathologically, pneumonia of any type was confirmed in 82 goats (70.7%) and CIP was present in 67 goats (57.8%). In most goats, the severity of the histopathological features of pneumonia was mild. Bacteria were detected in bronchial swabs from 73 goats (62.9%). CIP proved to be significantly positively linked to the occurrence of purulent bronchopneumonia (p < 0.001), fibrinous pleuropneumonia (p = 0.001), and of the infection of lungs with bacteria capable of causing pneumonia (p = 0.050). The causal character of these associations should be considered and warrants further investigation. Full article
(This article belongs to the Special Issue Emerging Infections in Small Ruminants)
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<p>The severity of three pneumonia types in a study goat population: chronic interstitial pneumonia (CIP), purulent bronchopneumonia (PBP), and fibrinous pleuropneumonia (FPP).</p>
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<p>The prevalence of three pneumonia types in a study goat population: chronic interstitial pneumonia (CIP), purulent bronchopneumonia (PBP), and fibrinous pleuropneumonia (FPP).</p>
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<p>Histopathological lesions observed in the chronic interstitial pneumonia (CIP): (<b>A</b>) lymphatic nodules hyperplasia (peribronchial and perivascular), interstitial (i.e., peribronchiolar and perialveolar) inflammatory infiltrates composed of macrophages and lymphocytes, interalveolar septum thickening, 100×, H-E; (<b>B</b>) interstitial (i.e., peribronchiolar and perialveolar) inflammatory infiltrates composed of macrophages and lymphocytes, interalveolar septum thickening, 200×, H-E; (<b>C</b>) smooth muscle hyperplasia in the walls of the bronchioles, 400×, H-E; and (<b>D</b>) interstitial connective tissue hyperplasia, 200×, Masson.</p>
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<p>Histopathological lesions observed in the purulent bronchopneumonia (PBP), 400×, H-E: (<b>A</b>) peribronchial inflammatory infiltrates composed of neutrophils and lymphocytes, purulent exudate in the lumen of bronchus; (<b>B</b>) focal necrosis of lung parenchyma; and (<b>C</b>) focal atelectasis.</p>
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<p>Histopathological lesions observed in the fibrinous pleuropneumonia pattern (FPP): (<b>A</b>) subpleural accumulation of fibrin, 200×, H-E; (<b>B</b>) subpleural accumulation of fibrin, 400×, Weigert; and (<b>C</b>) subpleural connective tissue hyperplasia, 400×, Masson.</p>
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<p>Representative images of immunohistochemistry (IHC) examination of caprine lung samples. The immunoreactivity for small ruminant lentivirus (SRLV) was developed with DAB chromogen (brown precipitate in the cell’s cytoplasm) and counterstained with Mayer’s hematoxylin (<b>A</b>–<b>D</b>) positive immunostaining and (<b>E</b>,<b>F</b>) negative controls: (<b>A</b>) the bronchiole lined by SRLV-positive epithelial cells (black arrowheads), 200×; (<b>B</b>) the SRLV-positive bronchial epithelial cells (black arrowhead) and a macrophage (white arrowhead) in the peribronchial inflammatory cells, 400×; (<b>C</b>) the SRLV-positive macrophages scattered in the thickened alveolar septa (white arrowheads), 200×; (<b>D</b>) the positive immunostaining of SRLV in the cytoplasm of the bronchial epithelial cells (black arrowhead) and macrophages (white arrowhead), 400×; (<b>E</b>) a negative tissue control, 200×; and (<b>F</b>) an isotype control, 200×.</p>
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6 pages, 544 KiB  
Communication
Serological Survey of Small Ruminant Lentivirus Infections in Free-Ranging Mouflon and Chamois in Slovenia
by Urška Kuhar, Diana Žele Vengušt and Gorazd Vengušt
Animals 2022, 12(8), 1032; https://doi.org/10.3390/ani12081032 - 15 Apr 2022
Cited by 3 | Viewed by 1825
Abstract
Small ruminant lentiviruses (SRLVs) belong to the genus Lentivirus in the Retroviridae family, which are responsible for the diseases maedi-visna and caprine arthritis-encephalitis in sheep and goats worldwide and are also widespread in Slovenian sheep and goats. SRLVs cause lifelong infections with chronic [...] Read more.
Small ruminant lentiviruses (SRLVs) belong to the genus Lentivirus in the Retroviridae family, which are responsible for the diseases maedi-visna and caprine arthritis-encephalitis in sheep and goats worldwide and are also widespread in Slovenian sheep and goats. SRLVs cause lifelong infections with chronic inflammatory lesions in various organ systems. Cross-species transmission of SRLV strains in sheep and goats is well documented, but there are few data on the ability of these viruses to infect wild ruminants. The objective of this study was to investigate whether SRLVs circulate among wild small ruminants in Slovenia. During the 2017–2018 hunting season, a total of 38 blood samples were collected from free-ranging chamois (Rupicapra rupicapra) and European mouflon (Ovis ammon musimon). The serum samples were tested for antibodies against SRLV by enzyme-linked immunosorbent assay (ELISA). The serological tests revealed that of all tested mouflons, 1 animal (11.1%) was seropositive, while all samples from chamois were negative. Based on the results of this study and considering the results of previous studies in which SRLV infections were detected in mouflons with low seroprevalence, it is very likely that the detected seropositive animal was an incidental spillover host for SRLV. Although no seropositive samples were found in chamois, we cannot speculate on whether chamois may not be a host for SRLV infection because of the small sample size and the disadvantages of the ELISA assay used when applied to samples from chamois. Full article
(This article belongs to the Special Issue Animals Viruses)
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<p>Geographical locations of SRLV antibody-negative and -positive samples from chamois and mouflon in Slovenia tested with ELISA.</p>
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23 pages, 4261 KiB  
Article
Molecular Characterization of Small Ruminant Lentiviruses Isolated from Polish Goats with Arthritis
by Monika Olech, Anna Kycko and Jacek Kuźmak
Viruses 2022, 14(4), 735; https://doi.org/10.3390/v14040735 - 31 Mar 2022
Cited by 6 | Viewed by 1893
Abstract
Previous studies revealed that the small ruminant lentivirus (SRLV) population in Poland is highly heterogeneous. All SRLVs detected from Polish sheep and goats so far have belonged to subtypes B1, B2, A1, A5, A12, A13, A16, A17, A18, A23 and A24. However, all [...] Read more.
Previous studies revealed that the small ruminant lentivirus (SRLV) population in Poland is highly heterogeneous. All SRLVs detected from Polish sheep and goats so far have belonged to subtypes B1, B2, A1, A5, A12, A13, A16, A17, A18, A23 and A24. However, all characterized strains originated from asymptomatic animals. This is the first study that characterizes the molecular properties of SRLVs isolated from different organs of six arthritic goats. Segments from three genomic regions (gag, LTR and env) were analyzed. In addition, we quantified the SRLV proviral load in the blood and different organs and examined its association with different degrees of histopathological lesions. All sequences obtained from the goats involved in this study were homogeneous, showing an average degree of variability of 4.8%, 3.7% and 8.8% for gag, LTR and env, respectively. Phylogenetic analysis revealed that the sequences from the analyzed goats were clustered within SRLVs group A and formed a new subtype within this group, tentatively named A27. The histopathological examination of the lung, mammary gland, synovial membranes of joints and brain of the analyzed goats revealed evidence of inflammatory processes associated with SRLV infection, which was confirmed by positive immunohistochemistry assays. No significant correlation was observed between histological features and alterations in the sequences from different tissues. No tissue-specific signature pattern was identified. It was shown that animals with a higher proviral load showed more lesion severity in various SRLV-affected tissues, indicating a positive association between these two parameters. Our results also revealed differences in the SRLV load between animals even though the sequences derived from all of the goats were closely related, suggesting that host factors may restrict and control viral replication. This study provides new information about SRLV variants isolated from arthritic goats; however, more studies, including the isolation and characterization of biological properties of these viruses, should be performed to evaluate their pathogenic potential. Full article
(This article belongs to the Special Issue State-of-the-Art Animal Virus Research in Poland)
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<p>Example photomicrographs of tissue sections from the CAEV-infected goats. (<b>A</b>–<b>C</b>,<b>E</b>,<b>G</b>)—Sections stained with hematoxylin and eosin (HE); (<b>D</b>,<b>F</b>,<b>H</b>)—Immunohistochemical (IHC) labeling for CAEV p28 detection. (<b>A</b>) Brainstem, goat 5. Locally present mononuclear infiltrations around the blood vessel; (<b>B</b>) Choroid plexus, goat 5. Mononuclear infiltration within the choroid; (<b>C</b>) Lung, goat 7. Moderate interstitial pneumonia. The alveolar septa and peribronchial interstitium are widened by diffuse mononuclear infiltration and eosinophilic amorphic material; (<b>D</b>) Lung, goat 7. Positive CAEV p28 immunolabeling visible as brown particles in the cytoplasm of the bronchiolar epithelium (black arrows) and single macrophage-like cells (white arrows); (<b>E</b>) Carpal joint, goat 5. Hypertrophic synovial membrane-forming villous projections thickened by mononuclear infiltrates; (<b>F</b>) Carpal joint, goat 7. Positive CAEV p28 immunolabeling visible as brown particles in the cytoplasm of singular cells (arrows) within subsynovial mononuclear infiltrations; (<b>G</b>) Mammary gland, goat 5. Multifocal to diffuse mononuclear infiltrations around lactiferous ducts, blood vessels and within interstitial tissue; (<b>H</b>) Mammary gland, goat 5. Positive CAEV p28 immunolabeling in the cytoplasm of macrophage-like cells infiltrating the periacinar area; Scale bars = 50 µm.</p>
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<p>Maximum-likelihood phylogenetic tree based on the alignment of the CA fragment of the <span class="html-italic">gag</span> gene. Sequences from this study are labeled by black circles. Numbers at the branches indicate the percentage of bootstrap values obtained from 1000 replicates. Bootstrap values &gt; 70% are shown. In the present study, the SRLV subtypes found by Colitti et al. [<a href="#B23-viruses-14-00735" class="html-bibr">23</a>] were renamed from A18 to A19 and from A19 to A20, and the subtypes found by Olech et al. [<a href="#B8-viruses-14-00735" class="html-bibr">8</a>] were renamed from A23 to A25 and from A24 to A26; goat 2—Blue, goat 3—Black, goat 4—Yellow, goat 5—Green, goat 6—Red, goat 7—Pink.</p>
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<p>Maximum-likelihood phylogenetic tree based on the alignment of the V4V5 fragment of the <span class="html-italic">env</span> gene. Sequences from this study are labeled by black circles. Numbers at the branches indicate the percentage of bootstrap values obtained from 1000 replicates. Bootstrap values &gt;70% are shown. In the present study, the SRLV subtypes found by Olech et al. [<a href="#B8-viruses-14-00735" class="html-bibr">8</a>] were renamed from A23 to A25 and from A24 to A26.</p>
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<p>Alignment (MUSCLE) of deduced amino acid sequences of the <span class="html-italic">gag</span>-p25 capsid protein of SRLVs obtained in this study and K1514 (GenBank accession number M60609) and Cork (GenBank accession number M33677) reference strains, which are MVV (group A) and CAEV (group B) prototype strains, respectively. Identical residues are indicated by dots (.). Two immunodominant epitopes and the major homology region (MHR) are within squares. PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland.</p>
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<p>Alignment of deduced amino acid sequences of an immunodominant epitope of the ENV protein and variable region V4 of SRLVs obtained in this study and reference strains. Deletions are indicated by a dash (-), and identical residues are indicated by dots (.). PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland.</p>
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<p>Maximum-likelihood phylogenetic tree based on the alignment of the LTR fragment. Sequences from this study are labeled by black circles. Numbers at the branches indicate the percentage of bootstrap values obtained from 1000 replicates. Bootstrap values &gt;70% are shown. PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland, goat 2—Blue, goat 3—Black, goat 4—Yellow, goat 5—Green, goat 6—Red, goat 7—Pink.</p>
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<p>Alignment of U3-R sequences of LTR region from Polish SRLV strains. Sequences are aligned against prototype strains K1514 and Cork representative of SRLV groups A and B, respectively. Dots indicate identity with Cork, and dashes represent gaps. Boundaries between U3, R and U5 are indicated by straight arrows. AP-1, AP-4, AML (vis) motifs, TATA box, GAS, TAS and polyadenylation signal (poly A) are marked by boxes. Gray boxes represent CAAAT sequences. PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland.</p>
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<p>Alignment of U3-R sequences of LTR region from Polish SRLV strains. Sequences are aligned against prototype strains K1514 and Cork representative of SRLV groups A and B, respectively. Dots indicate identity with Cork, and dashes represent gaps. Boundaries between U3, R and U5 are indicated by straight arrows. AP-1, AP-4, AML (vis) motifs, TATA box, GAS, TAS and polyadenylation signal (poly A) are marked by boxes. Gray boxes represent CAAAT sequences. PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland.</p>
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<p>Quantification of SRLV proviral load in different organs and cells of goats analyzed in this study. PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland. Mean value of SRLV proviral copy number is shown above the bar.</p>
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<p>Quantification of SRLV proviral load in different organs and cells of goats analyzed in this study. PBMC—Peripheral blood mononuclear cells, L—Lung, SM—Synovial membrane, B—Brain, CHP—Choroid plexus, MG—Mammary gland. Mean value of SRLV proviral copy number is shown above the bar.</p>
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13 pages, 1971 KiB  
Article
Genetic Characterization of Small Ruminant Lentiviruses (SRLVs) Circulating in Naturally Infected Sheep in Central Italy
by Chiara Arcangeli, Martina Torricelli, Carla Sebastiani, Daniele Lucarelli, Marcella Ciullo, Fabrizio Passamonti, Monica Giammarioli and Massimo Biagetti
Viruses 2022, 14(4), 686; https://doi.org/10.3390/v14040686 - 25 Mar 2022
Cited by 12 | Viewed by 2507
Abstract
Small ruminant lentiviruses (SRLVs) represent a very heterogeneous group of ss-RNA viruses that infect sheep and goats worldwide. They cause important, deleterious effects on animal production and limit the animal trade. SRLVs show a high genetic variability due to high mutation rate and [...] Read more.
Small ruminant lentiviruses (SRLVs) represent a very heterogeneous group of ss-RNA viruses that infect sheep and goats worldwide. They cause important, deleterious effects on animal production and limit the animal trade. SRLVs show a high genetic variability due to high mutation rate and frequent recombination events. Indeed, five genotypes (A–E) and several subtypes have been detected. The aim of this work was to genetically characterize SRLVs circulating in central Italy. On this basis, a phylogenetic study on the gag-pol genetic region of 133 sheep, collected from 19 naturally infected flocks, was conducted. In addition, to evaluate the frequency of mutation and the selective pressure on this region, a WebLogo 3 analysis was performed, and the dN/dS ratio was computed. The results showed that 26 samples out of 133 were clustered in genotype A and 106 samples belonged to genotype B, as follows: A9 (n = 8), A11 (n = 10), A24 (n = 7), B1 (n = 2), B2 (n = 59), and B3 (n = 45). No recombination events were found. Mutations were localized mainly in the VR-2 region, and the dN/dS ratio of 0.028 indicated the existence of purifying selection. Since the genetic diversity of SRLVs could make serological identification difficult, it is important to perform molecular characterization to ensure a more reliable diagnosis, to maintain flock health status, and for the application of local and national control programs. Full article
(This article belongs to the Topic Veterinary Infectious Diseases)
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<p>Maximum likelihood phylogenetic tree based on the alignment of 642 nt from <span class="html-italic">gag-pol</span> region of 287 sequences: 133 analysed in this study (labeled by a different color for each subtype) and 154 reference strains available in GenBank (labeled by black color). Bar: number of substitutions per site. Correspondence between sample names and accession numbers are reported in <a href="#app1-viruses-14-00686" class="html-app">Table S1</a>.</p>
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<p>Amino acid sequence multiple alignment of SRLVs deduced from the <span class="html-italic">gag-pol</span> fragment. Each subtype has been aligned with the respective reference sequence. Immunodominant epitopes 2 and 3, major homology region (MHR) and variable region 2 (VR-2) are reported. Dots represent the same amino acid residue. Correspondence between sample names and accession numbers are reported in <a href="#app1-viruses-14-00686" class="html-app">Table S1</a>. (<b>a</b>) Subtypes A9, A11, A24, B1, B2; (<b>b</b>) Subtype B3.</p>
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<p>Amino acid sequence multiple alignment of SRLVs deduced from the <span class="html-italic">gag-pol</span> fragment. Each subtype has been aligned with the respective reference sequence. Immunodominant epitopes 2 and 3, major homology region (MHR) and variable region 2 (VR-2) are reported. Dots represent the same amino acid residue. Correspondence between sample names and accession numbers are reported in <a href="#app1-viruses-14-00686" class="html-app">Table S1</a>. (<b>a</b>) Subtypes A9, A11, A24, B1, B2; (<b>b</b>) Subtype B3.</p>
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<p>Graphical representation of the relative amino acid frequency of the partial SRLV <span class="html-italic">gag-pol</span> protein obtained by WebLogo 3 software. The height of the letter corresponding to each amino acid indicates its relative frequency at that specific position. Different colors indicate the physiochemical characteristics of the amino acid (black: non-polar, green: polar, red: aromatic, blue: positively charged, purple: negatively charged).</p>
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29 pages, 5442 KiB  
Article
Species-Specific Humoral Immune Responses in Sheep and Goats upon Small Ruminant Lentivirus Infections Inversely Correlate with Protection against Virus Replication and Pathological Lesions
by Rodolphe Michiels, Stefan Roels, Nick Vereecke, Elisabeth Mathijs, Laurent Mostin and Nick De Regge
Int. J. Mol. Sci. 2021, 22(18), 9824; https://doi.org/10.3390/ijms22189824 - 11 Sep 2021
Cited by 4 | Viewed by 1839
Abstract
Maedi-Visna-like genotype A strains and Caprine arthritis encephaltis-like genotype B strains are small ruminant lentiviruses (SRLV) which, for incompletely understood reasons, appear to be more virulent in sheep and goats, respectively. A 9-month in vivo infection experiment using Belgian genotype A and B [...] Read more.
Maedi-Visna-like genotype A strains and Caprine arthritis encephaltis-like genotype B strains are small ruminant lentiviruses (SRLV) which, for incompletely understood reasons, appear to be more virulent in sheep and goats, respectively. A 9-month in vivo infection experiment using Belgian genotype A and B SRLV strains showed that almost all homologous (genotype A in sheep; genotype B in goats) and heterologous (genotype A in goats; genotype B in sheep) intratracheal inoculations resulted in productive infection. No differences in viremia and time to seroconversion were observed between homologous and heterologous infections. Higher viral loads and more severe lesions in the mammary gland and lung were however detected at 9 months post homologous compared to heterologous infection which coincided with strongly increased IFN-γ mRNA expression levels upon homologous infection. Pepscan analysis revealed a strong antibody response against immune-dominant regions of the capsid and surface proteins upon homologous infection, which was absent after heterologous infection. These results inversely correlated with protection against virus replication in target organs and observed histopathological lesions, and thus require an in-depth evaluation of a potential role of antibody dependent enhancement in SRLV infection. Finally, no horizontal intra- and cross-species SRLV transmission to contact animals was detected. Full article
(This article belongs to the Section Molecular Immunology)
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<p>SRLV RNA detection in target organs of sheep (SH) and goats (G) at 9 months after homologous and heterologous genotype A (panel <b>A</b>) and B (panel <b>B</b>) strain infection. Bars indicate the number of qPCR positive animals. Each dot corresponds to one animal and represents the viral load. The mean viral load is indicated by the horizontal line.</p>
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<p>Representative histopathological lesions in the mammary gland of genotype A and B SRLV strain infected sheep and goats at 9 months <span class="html-italic">p.i</span>. (<b>A</b>) Section from an uninfected sheep exhibiting structured acini and no lymphocyte infiltration. (<b>B</b>) Section from a sheep infected with a genotype A strain exhibiting a massive lymphocyte infiltration (asterisks) and a destruction of the acini structure (arrow). (<b>C</b>) Section from an uninfected goat exhibiting tissues with normal glandular structures and no lymphocytic infiltrate. (<b>D</b>) Section from a goat infected with a genotype B strain exhibiting lesions with massive lymphocyte infiltrations and an atrophy of the acini. Magnification ×20.</p>
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<p>Cytokines’ mRNA expression in the mammary gland of genotype A and genotype B infected sheep and goats. mRNA expression was detected by qPCR for IFN-α, IFN-β, IFN-γ, TNF-α, IL-8, IL-10, TGF-β1 and GM-CSF.</p>
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<p>Cytokines’ mRNA expression in the lung of genotype A and genotype B infected sheep and goats. mRNA expression was detected by qPCR for IFN-α, IFN-β, IFN-γ, TNF-α, IL-8, IL-10, TGF-β1 and GM-CSF.</p>
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<p>Cytokines’ mRNA expression in the cerebellum of genotype A and genotype B infected sheep and goats. mRNA expression was detected by qPCR for IFN-α, IFN-β, IFN-γ, TNF-α, IL-8, IL-10, TGF-β1 and GM-CSF.</p>
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<p>Pepscan analysis of sera from genotype A infected sheep and goats against linear peptides of the capsid and surface proteins of the genotype A strain used as inoculum. Sera were tested at a dilution of 1/100, and each bar represents the reactivity of one serum against one single peptide.</p>
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<p>Pepscan analysis of sera from genotype B infected sheep and goats against linear peptides of the capsid and surface proteins of the genotype B strain used as inoculum. Sera were tested at a dilution of 1/100, and each bar represents the reactivity of one serum against one single peptide.</p>
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<p>Detection of specific antibodies over time in sera collected from genotype A infected sheep and goats against selected epitopes of the capsid and surface proteins via pepscan analysis. Sera collected at 1, 3, 6 and 9 months (m) <span class="html-italic">p.i.</span> were tested at a dilution of 1/100, and each bar represents the reactivity of one serum against one single peptide.</p>
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<p>Detection of specific antibodies over time in sera collected from genotype B infected sheep and goats against selected epitopes of the capsid and surface proteins via pepscan analysis. Sera collected at 1, 3, 6 and 9 months <span class="html-italic">p.i.</span> were tested at a dilution of 1/100, and each bar represents the reactivity of one serum against one single peptide.</p>
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12 pages, 296 KiB  
Article
First Survey of SNPs in TMEM154, TLR9, MYD88 and CCR5 Genes in Sheep Reared in Italy and Their Association with Resistance to SRLVs Infection
by Chiara Arcangeli, Daniele Lucarelli, Martina Torricelli, Carla Sebastiani, Marcella Ciullo, Claudia Pellegrini, Andrea Felici, Silva Costarelli, Monica Giammarioli, Francesco Feliziani, Fabrizio Passamonti and Massimo Biagetti
Viruses 2021, 13(7), 1290; https://doi.org/10.3390/v13071290 - 1 Jul 2021
Cited by 10 | Viewed by 2562
Abstract
Maedi-visna virus (MVV) and caprine arthritis encephalitis virus (CAEV), referred to as small ruminant lentiviruses (SRLVs), belong to the genus Lentivirus of the Retroviridae family. SRLVs infect both sheep and goats, causing significant economic losses and animal welfare damage. Recent findings suggest an [...] Read more.
Maedi-visna virus (MVV) and caprine arthritis encephalitis virus (CAEV), referred to as small ruminant lentiviruses (SRLVs), belong to the genus Lentivirus of the Retroviridae family. SRLVs infect both sheep and goats, causing significant economic losses and animal welfare damage. Recent findings suggest an association between serological status and allelic variants of different genes such as TMEM154, TLR9, MYD88 and CCR5. The aim of this work was to investigate the role of specific polymorphisms of these genes in SRLVs infection in some sheep flocks in Italy. In addition to those already known, novel variants in the TMEM154 (P7H, I74V, I105V) gene were detected in this study. The risk of infection was determined finding an association between the serological status and polymorphisms P7H, E35K, N70I, I74V, I105V of TMEM154, R447Q, A462S and G520R in TLR9 gene, H176H* and K190K* in MYD88 genes, while no statistical association was observed for the 4-bp deletion of the CCR5 gene. Since no vaccines or treatments have been developed, a genetically based approach could be an innovative strategy to prevent and to control SRLVs infection. Our findings are an important starting point in order to define the genetic resistance profile towards SRLVs infection. Full article
(This article belongs to the Special Issue Non-Primate Lentiviruses 2021)
17 pages, 4164 KiB  
Article
A Comparison of Oxidative Stress Biomarkers in the Serum of Healthy Polish Dairy Goats with Those Naturally Infected with Small Ruminant Lentivirus in the Course of Lactation
by Natalia Kurhaluk, Halyna Tkachenko, Michał Czopowicz, Jacek Sikora, Daria M. Urbańska, Aldona Kawęcka, Jarosław Kaba and Emilia Bagnicka
Animals 2021, 11(7), 1945; https://doi.org/10.3390/ani11071945 - 29 Jun 2021
Cited by 6 | Viewed by 2300
Abstract
The present study examines the effects of natural infection by small ruminant lentivirus (SRLV) in the two most common goat breeds in Poland, i.e., Polish white improved and polish fawn improved. It focuses on biomarkers of oxidative stress, oxidatively modified proteins and antioxidant [...] Read more.
The present study examines the effects of natural infection by small ruminant lentivirus (SRLV) in the two most common goat breeds in Poland, i.e., Polish white improved and polish fawn improved. It focuses on biomarkers of oxidative stress, oxidatively modified proteins and antioxidant defenses, ceruloplasmin level as an acute phase protein, and the activities of antioxidant enzymes in the goat serum. It was conducted on 24 goats divided into two equal groups: one SRLV-seropositive (SRLV-SP) and another SRLV-seronegative (SRLV-SN). Both groups were identical in terms of breed and parity. Despite infection, the SRLV-SP goats demonstrated no symptoms of caprine arthritis-encephalitis. In addition, the SRLV-SP goats did not reveal pronounced dysfunctions in oxidative stress biomarkers in the serum compared to the SRLV-SN animals. However, both groups demonstrated elevated levels of the aldehydic and ketonic derivatives of oxidatively modified proteins during the lactation period. In addition, both groups retained a high total antioxidant capacity in serum despite the decrease of enzyme antioxidant defenses throughout the 200-day lactation period. Full article
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<p>Lipid peroxidation, estimated by TBARS content (µmol MDA·L<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b, c, d—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’, c’, d’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Ketonic derivatives of oxidatively modified proteins (nmol·mL<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Aldehydic derivatives of oxidatively modified proteins (nmol·mL<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>The total antioxidant capacity (TAC, %) in the serum of SRLV-SN and and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>The total antioxidant capacity (TAC, %) in the serum of SRLV-SN and and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Ceruloplasmin level (Cp, mg·L<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b, c—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’, c’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Superoxide dismutase activity (SOD, U·mL<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b, c—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’, c’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Catalase activity (CAT, mmol H<sub>2</sub>O<sub>2</sub>·min<sup>−1</sup>·L<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b, c—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’, c’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Glutathione reductase activity (GR, μmol NADPH<sub>2</sub>·min<sup>−1·</sup>mL<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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<p>Glutathione peroxidase activity (μmol GSH·min<sup>−1</sup>·mL<sup>−1</sup>) in the serum of SRLV-SN and SRLV-SP goats during lactation. Values are expressed as means ± SEM; a, b, c—different letters indicate significant differences between stages of lactation within the SRLV-SN group at <span class="html-italic">p</span> &lt; 0.05; a’, b’, c’—different letters indicate significant differences between stages of lactation within the SRLV-SP group at <span class="html-italic">p</span> &lt; 0.05; SRLV-SP—small ruminant lentivirus seropositive goats; SRLV-SN—small ruminant lentivirus seronegative goats.</p>
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15 pages, 806 KiB  
Article
A DNA Regulatory Element Haplotype at Zinc Finger Genes Is Associated with Host Resilience to Small Ruminant Lentivirus in Two Sheep Populations
by Alisha T. Massa, Michelle R. Mousel, Codie J. Durfee, Maria K. Herndon, Kaneesha M. Hemmerling, J. Bret Taylor, Holly L. Neibergs and Stephen N. White
Animals 2021, 11(7), 1907; https://doi.org/10.3390/ani11071907 - 26 Jun 2021
Cited by 1 | Viewed by 2517
Abstract
Small ruminant lentivirus (SRLV) causes Maedi-Visna or Ovine Progressive Pneumonia in sheep and creates insidious livestock production losses. This retrovirus is closely related to human immunodeficiency virus and currently has no vaccines or cure. Genetic marker assisted selection for sheep disease resiliency presents [...] Read more.
Small ruminant lentivirus (SRLV) causes Maedi-Visna or Ovine Progressive Pneumonia in sheep and creates insidious livestock production losses. This retrovirus is closely related to human immunodeficiency virus and currently has no vaccines or cure. Genetic marker assisted selection for sheep disease resiliency presents an attractive management solution. Previously, we identified a region containing a cluster of zinc finger genes that had association with ovine SRLV proviral concentration. Trait-association analysis validated a small insertion/deletion variant near ZNF389 (rs397514112) in multiple sheep breeds. In the current study, 543 sheep from two distinct populations were genotyped at 34 additional variants for fine mapping of the regulatory elements within this locus. Variants were selected based on ChIP-seq annotation data from sheep alveolar macrophages that defined active cis-regulatory elements predicted to influence zinc finger gene expression. We present a haplotype block of variants within regulatory elements that have improved associations and larger effect sizes (up to 4.7-fold genotypic difference in proviral concentration) than the previously validated ZNF389 deletion marker. Hypotheses for the underlying causal mutation or mutations are presented based on changes to in silico transcription factor binding sites. These variants offer alternative markers for selective breeding and are targets for future functional mutation assays. Full article
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<p>Association with small ruminant lentivirus proviral concentration at variants within regulatory elements for zinc finger genes on ovine chromosome 20. ChIP-seq annotations for H3K4me3 enrichment are shown as green bars at the bottom, above Refseq gene annotations in black. Population 1 (purebred Rambouillet) are identified by blue dots and population 2 (crossbred Rambouillet, Columbia) are identified by orange dots. The previously validated marker (rs397514112) is highlighted by green circles for each population.</p>
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<p>Association with SRLV proviral concentration at higher resolution for variants within the two regulatory elements near ZNF389 that flank the previously validated insertion/deletion marker (rs397514112) on chromosome 20. Below, the ChIP-seq signal tracks for two animals enriched for H3K27ac (active proximal enhancers) and H3K4me3 (active promoters) is displayed. Transcription start site data from cap analysis gene expression (CAGE) from alveolar macrophages is also displayed [<a href="#B27-animals-11-01907" class="html-bibr">27</a>]. Peak calls significant in both animals are indicated by the solid blue or green bars. Refseq gene annotation is displayed in black along the bottom.</p>
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<p>Haplotypes and population frequency as determined by PHASE. On the left are the haplotypes within population 1 (purebred Rambouillet) and on the right are haplotypes for population 2 (crossbred, Rambouillet, Columbia). The same three haplotypes were the most common in both populations. The fifteen loci are shown in columns listed in the same order as in <a href="#animals-11-01907-t001" class="html-table">Table 1</a> and <a href="#animals-11-01907-t002" class="html-table">Table 2</a>, with the previously validated marker listed as +. An asterisk (*) is above each locus in the two key regulatory elements upstream of <span class="html-italic">ZNF389</span>. For loci 09 and 14, D represents the deletion allele and I represents the insertion allele.</p>
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11 pages, 604 KiB  
Article
Does Small Ruminant Lentivirus Infection in Goats Predispose to Bacterial Infection of the Mammary Gland? A Preliminary Study
by Daria Urbańska, Ryszard Puchała, Justyna Jarczak, Michał Czopowicz, Jarosław Kaba, Karina Horbańczuk and Emilia Bagnicka
Animals 2021, 11(7), 1851; https://doi.org/10.3390/ani11071851 - 22 Jun 2021
Cited by 5 | Viewed by 1826
Abstract
The aim of this study was to determine whether asymptomatic small ruminant lentivirus seropositive (SRLV-SP) goats were more susceptible to bacterial infection of the udder when lactating by comparing the presence and species of pathogenic bacteria in their milk with the values for [...] Read more.
The aim of this study was to determine whether asymptomatic small ruminant lentivirus seropositive (SRLV-SP) goats were more susceptible to bacterial infection of the udder when lactating by comparing the presence and species of pathogenic bacteria in their milk with the values for seronegative goats (SRLV-SN). Milk samples were collected during morning milking on days 20, 40, 60, 150, and 210 of lactation for three consecutive years and subjected to bacteriological examination. Staphylococcus caprae and S. xylosus were the most frequent strains identified in both SRLV-SP and SRLV-SN goats. The prevalence of pathogenic bacteria was the highest in the 1st lactation, regardless of SRLV status. Moreover, the prevalence of pathogenic bacteria was significantly higher in SRLV-SP goats, but only those in the 5th or further lactation (p = 0.010). This suggests a relationship between long-lasting SRLV infection and susceptibility to bacterial infections of the udder. Full article
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<p>Prevalence of bacteria in milk samples from subsequent lactations, regardless of small ruminant lentivirus infection status. The asterisk (*) indicates statistical significance at α = 0.05.</p>
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<p>Prevalence of bacteria in milk samples from small ruminant lentivirus-seropositive (SRLV-SP) and seronegative (SRLV-SN) goats in subsequent lactations. Whiskers indicate CI 95% and the asterisk (*) indicates statistical significance at α = 0.05.</p>
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Article
Molecular Characterization of Small Ruminant Lentiviruses of Subtype A5 Detected in Naturally Infected but Clinically Healthy Goats of Carpathian Breed
by Monika Olech and Jacek Kuźmak
Pathogens 2020, 9(12), 992; https://doi.org/10.3390/pathogens9120992 - 26 Nov 2020
Cited by 13 | Viewed by 2068
Abstract
Small ruminant lentiviruses (SRLVs) are widespread in sheep and goats in Poland, and several subtypes were identified and molecularly characterized up to date. This is the first study that characterizes the molecular properties of A5 strains of SRLV detected in naturally infected, but [...] Read more.
Small ruminant lentiviruses (SRLVs) are widespread in sheep and goats in Poland, and several subtypes were identified and molecularly characterized up to date. This is the first study that characterizes the molecular properties of A5 strains of SRLV detected in naturally infected, but clinically healthy, Carpathian goats. Segments from three genomic regions (gag, env, and LTR) were analyzed. Genetic distance, pairwise comparison, and phylogenetic analysis revealed that Polish SRLV A5 sequences are closely related to the Swiss and German A5 sequences suggesting a common origin. The epidemiological linkage was identified particularly between the small ruminants of Germany and Poland. Amino acid sequences of immunodominant regions in CA protein were well-conserved within analyzed strains; however, they showed some remarkable changes like substitution (D) to (E), at position 90 in Major Homology Region (MHR) and (T) to (S), at position 141 in epitope 3. In contrast, aa sequences of surface glycoprotein exhibited the highest variability confirming type-specific variation in SU5 epitope. Two deletions in the U3 region of A5 strains were noted: One (8 nt) located near the 5′ end of the U3 region and the other (29 nt) located in the central region of U3. Additionally, all A5 strains had specific deletion (10 nt) in the R region. Furthermore, we did not find a correlation between copies of the CAAAT motif and clinical manifestation in infected animals. These data showed some remarkable features in the viral genome of A5 strains, which may be related to the attenuated phenotype in vivo, characterized by the lack of any clinical signs in infected goats. Certainly, more studies are required to support the hypothesis that these A5 viruses are of low pathogenicity for goats. We want to focus our future studies on the analysis of the whole genomes of these isolates and their biological properties, as well as on clinicopathological studies of goats infected by A5 SRLV, aiming to clarify the pathogenic potential of these viruses. Full article
(This article belongs to the Section Viral Pathogens)
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<p>Maximum-likelihood phylogenetic tree based on the alignment of <span class="html-italic">gag</span> sequences (423 bp) from 77 sequences: Nine analyzes in this study sequences (labeled by black circles), and 69 reference small ruminant lentiviruses (SRLVs) strains originated from different geographical areas. Strains are shown by name followed by GenBank accession number, genotype, and country of origin. Numbers at the branches indicate the percentage of bootstrap values obtained from 1000 replicates. Bootstrap values &gt; 50% are shown.</p>
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<p>Pairwise percent identity of <span class="html-italic">gag</span> nucleotide and amino acid sequences of Polish SRLV A5 sequences compared to different subtypes of genotype A. The lower matrix shows the percent identity of nucleotide sequences, and the upper matrix shows the percent identity of amino acid sequences. The intensity of the color is correlated with pairwise percent identity.</p>
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<p>Sequence homology of Gag (<b>A</b>) and Env (<b>B</b>) immunodominant regions between Polish SRLV. A5 strains were analyzed in this study and the reference strains from different genotypes. Deletions are indicated by a dash (-), and identical residues are indicated by dots (.).</p>
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<p>Alignment of U3-R sequences of the long terminal repeats (LTR) region from Polish A5 SRLV strains. Sequences were aligned against virulent B1 strain Cork, neurovirulent A2/A3 strain 697, neurovirulent A1 strain Kv1772, arthritic B2 strain 496, and attenuated A4 strain 6221. Dots indicate identity with Cork, and dashes represent gaps. The boundaries between U3, R, and U5 are indicated by straight arrows. AP-1, AP-4, AML (vis) motifs, the TATA box, and polyadenylation signal (poly A) are marked by boxes. The grey boxes represent CAAAT sequences.</p>
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