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1.
Virus Res ; 336: 199209, 2023 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-37633596

RESUMEN

Classical swine fever virus (CSFV) shares high antigenic homology with other members of the genus Pestivirus. Because several pestivirus species can also infect swine, eliciting cross-reactive antibodies, it is important to define CSFV-specific epitopes for the differential diagnosis of classical swine fever (CSF) by serology. For this purpose, epitope mapping of seven monoclonal antibodies (mAbs), recognizing sites on the D/A domain of glycoprotein E2, was performed using recombinant expressed antigenic domains and mutants of E2, as well as an overlapping peptide library. Three CSFV-specific epitopes, i.e., 780-IEEMGDDFGFGLCPF-794, 810-NGSAFYLVCPIGWTG-824, and 846-REKPF-850, were identified within the D/A domain of E2. Site-directed mutagenesis further confirmed that residues 783-MGD-785, 789-FGLCPF-794, 813-AFYLVCPIGWTG-824, and 846-REK-848 were critical residues in these regions. In addition, a F789S difference within the epitope 780-IEEMGDDFGFGLCPF-794 was responsible for the absence of binding of two mAbs to the E2 protein of the live attenuated CSFV vaccine strain Riems. Structural modeling revealed that, the three epitopes are located near each other, suggesting that they may form a more complex conformational epitope on the D/A domain in vivo. Six of the mAbs neutralized viruses of diverse genotypes, indicating that the target epitopes are involved in virus interaction with cells. The binding of CSFV to cells was significantly reduced after pre-incubation with either truncated E2 proteins comprising the D/A domain or with the CSFV-specific mAbs targeting the domain D/A. These epitopes identified on the D/A domain are important targets for virus neutralization that might be involved in the early steps of CSFV infection. These findings reveal potential candidates for improving the differential diagnosis of pestiviruses by serology.

2.
Animals (Basel) ; 13(3)2023 Jan 21.
Artículo en Inglés | MEDLINE | ID: mdl-36766258

RESUMEN

Classical swine fever (CSF) is a systemic hemorrhagic disease affecting domestic pigs and wild boars. The modified live vaccine (MLV) induces quick and solid protection against CSF virus (CSFV) infection. Maternally derived antibodies (MDAs) via colostrum could interfere with the MLV's efficacy, leading to incomplete protection against CSFV infection for pigs. This study investigated CSFV transmission among experimental piglets with various post-MLV immune statuses. Nineteen piglets, 18 with MDAs and 1 specific-pathogen-free piglet infected with CSFV that served as the CSFV donor, were cohabited with piglets that had or had not been administered the MLV. Five-sixths of the piglets with MDAs that had been administered one dose of MLV were fully protected from contact transmission from the CSFV donor and did not transmit CSFV to the piglets secondarily exposed through cohabitation. Cell-mediated immunity, represented by the anti-CSFV-specific interferon-γ-secreting cells, was key to viral clearance and recovery. After cohabitation with a CSFV donor, the unvaccinated piglets with low MDA levels exhibited CSFV infection and spread CSFV to other piglets through contact; those with high MDA levels recovered but acted as asymptomatic carriers. In conclusion, MLV still induces solid immunity in commercial herds under MDA interference and blocks CSFV transmission within these herds.

3.
Viruses ; 13(8)2021 08 20.
Artículo en Inglés | MEDLINE | ID: mdl-34452520

RESUMEN

Classical swine fever virus (CSFV) shares high structural and antigenic homology with bovine viral diarrhea virus (BVDV) and border disease virus (BDV). Because all three viruses can infect swine and elicit cross-reactive antibodies, it is necessary to differentiate among them with regard to serological diagnosis of classical swine fever. To understand the mechanism of cross-reactivity, it is important to define common or specific epitopes of these viruses. For this purpose, epitope mapping of six monoclonal antibodies (mAbs) was performed using recombinant expressed antigenic domains of CSFV and BDV E2 proteins. One CSFV-specific conformational epitope and one CSFV and BDV common epitope within domain B/C of E2 were identified. Site-directed mutagenesis confirmed that residues G725 and V738/I738 of the CSFV-specific epitope and P709/L709 and E713 of the second epitope are important for mAbs binding. Infection of CSFV in porcine cells was significantly reduced after pre-incubation of the cells with the domain B/C of E2 or after pre-incubation of CSFV with the mAbs detecting domain B/C. 3D structural modeling suggested that both epitopes are exposed on the surface of E2. Based on this, the identified epitopes represent a potential target for virus neutralization and might be involved in the early steps of CSFV infection.


Asunto(s)
Enfermedad de la Frontera/virología , Virus de la Enfermedad de la Frontera/inmunología , Virus de la Fiebre Porcina Clásica/inmunología , Peste Porcina Clásica/virología , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/inmunología , Animales , Virus de la Enfermedad de la Frontera/química , Virus de la Enfermedad de la Frontera/genética , Virus de la Fiebre Porcina Clásica/química , Virus de la Fiebre Porcina Clásica/genética , Mapeo Epitopo , Epítopos/química , Epítopos/genética , Epítopos/inmunología , Dominios Proteicos , Porcinos , Enfermedades de los Porcinos/virología , Proteínas del Envoltorio Viral/genética
4.
Pathogens ; 10(1)2020 Dec 22.
Artículo en Inglés | MEDLINE | ID: mdl-33375108

RESUMEN

Bluetongue is an infectious viral hemorrhagic disease of domestic and wild ruminants that has a considerable economic impact on domestic ruminants. There are currently at least 29 serotypes of bluetongue virus (BTV) in the world. Noteworthily, the pathogenesis among BTV serotypes is different, even in the same animal species. In this study, BTV2/KM/2003 and BTV12/PT/2003 were used to investigate the differential immunological effects on bovine peripheral blood mononuclear cells (PBMCs). The BTV viral load and the expression of cytokine messenger RNA (mRNA) in PBMCs were measured by fluorescence-based real-time reverse-transcription PCR (qRT-PCR). The immunofluorescence assay (IFA) was applied to detect BTV signals in monocyte-derived macrophages (MDMs). The SWISS-MODEL and IL-4pred prediction tools were used to predict the interleukin 4 (IL-4)-inducing peptides in BTV-coat protein VP2. Synthetic peptides of VP2 were used to stimulate PBMCs for IL-4-inducing capability. This study demonstrated that the cytokine profiles of BTV-induced PBMCs were significantly different between BTV2/KM/2003 and BTV12/PT/2003. BTV2 preferentially activated the T helper 2 (Th2) pathway, represented by the early induction of IL-4, and likely fed back to inhibit the innate immunity. In contrast, BTV12 preferentially activated the innate immunity, represented by the induction of tumor necrosis factor -α (TNF-α) and interleukin 1 (IL-1), with only minimal subsequent IL-4. The BTV nonstructural protein 3 antibody (anti-BTV-NS3) fluorescent signals demonstrated that monocytes in PBMCs and MDMs were the preferred targets of BTV replication. Bioinformatics analysis revealed that the capability to induce IL-4 was attributed to the tip region of the VP2 protein, wherein a higher number of predicted peptide segments on BTVs were positively correlated with the allergic reaction reported in cattle. Synthetic peptides of BTV2-VP2 induced significant IL-4 within 12-24 h post-infection (hpi) in PBMCs, whereas those of BTV12 did not, consistent with the bioinformatics prediction. Bovine PBMCs and synthetic peptides together seem to serve as a good model for pursuing the BTV-induced IL-4 activity that precedes the development of an allergic reaction, although further optimization of the protocol is warranted.

5.
Viruses ; 12(12)2020 12 02.
Artículo en Inglés | MEDLINE | ID: mdl-33276502

RESUMEN

Porcine epidemic diarrhea virus (PEDV) causes severe diarrhea and a high rate of mortality in suckling pigs. The epidemic of PEDV that occurred after 2013 was caused by non-insertion and deletion of S gene (S-INDEL) PEDV strains. During this epidemic, a variant of the non-S-INDEL PEDV strain with a large deletion of 205 amino acids on the spike gene (5-17-V) was also found to co-exist with a non-S-INDEL PEDV without deletion (5-17-O). Herein, we describe the differences in the complete genome, distribution, virulence, and antigenicity between strain 5-17-O and variant strain 5-17-V. The deletion of 205 amino acids was primarily located in the S1O domain and was associated with milder clinical signs and lower mortality in suckling pigs than those of the 5-17-O strain. The 5-17-V strain-induced antibody did not completely cross-neutralize the 5-17-O strain. In conclusion, the deletion in the S1 region reduces the virulence of PEDV and influences the virus-neutralizing activities of the antibody it induces.


Asunto(s)
Anticuerpos Antivirales/inmunología , Formación de Anticuerpos/inmunología , Infecciones por Coronavirus/veterinaria , Interacciones Huésped-Patógeno/inmunología , Virus de la Diarrea Epidémica Porcina/fisiología , Eliminación de Secuencia , Glicoproteína de la Espiga del Coronavirus/genética , Secuencia de Aminoácidos , Animales , Anticuerpos Neutralizantes/inmunología , Diarrea/veterinaria , Genoma Viral , Genómica/métodos , Pruebas de Neutralización , Variantes Farmacogenómicas , Filogenia , Virus de la Diarrea Epidémica Porcina/clasificación , Porcinos , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/virología , Taiwán , Virulencia/genética , Secuenciación Completa del Genoma
6.
Viruses ; 12(11)2020 10 29.
Artículo en Inglés | MEDLINE | ID: mdl-33138189

RESUMEN

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


Asunto(s)
Biología Computacional , Mapeo Epitopo/métodos , Epítopos/genética , Epítopos/inmunología , Sueros Inmunes , Teschovirus/inmunología , Animales , Anticuerpos Neutralizantes/inmunología , Simulación por Computador , Femenino , Ratones , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Serogrupo , Porcinos , Enfermedades de los Porcinos/virología , Teschovirus/patogenicidad
7.
Pathogens ; 9(9)2020 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-32927731

RESUMEN

Recent reemergence of classical swine fever (CSF) in previous CSF-free areas reminds the veterinary community of this old disease [...].

8.
Vet Immunol Immunopathol ; 226: 110071, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32540689

RESUMEN

Bluetongue is a fatal viral disease in ruminants and has serious economic impacts on the livestock industry. Interactions between bluetongue virus (BTV) and immune cells are interesting because of the unique scenarios in each combination of animal species/breed and viral virulence/serotype. This study investigated the immune response in bovine peripheral blood mononuclear cells (PBMC) infected by the BTV2 Taiwan strain. The replication of the virus was limited in monocytes and monocyte-derived macrophages (MDM), and lymphocytes were less permissive. The cytokine mRNA of IL-4 in PBMC was expressed earlier and in greater quantities than that of innate immunity (TNFα, IL-1ß) and cell mediated immunity (CMI) (IFNγ), and the IL-4 protein was stably present in the culture medium until 72 h post-infection (hpi). Even in MDM reconstituted with autologous lymphocyte (MDM-Lymphocyte), the IL-4 still had high mRNA expression level. The level of IgE antibody also increased at 24-72 hpi, suggestive of the engagement of type I hypersensitivity in the pathogenesis. The anti-viral activity contained in the culture supernatant was transferrable to recipient infected PBMC from other cows. However, in infected MDM largely free of lymphocytes, mRNA expressions of IL-1ß, TNFα and IL-12p40 were normally expressed from 6 to 48 hpi, supporting the notion that IL-4 elaborated by lymphocytes in PBMC mediated the inhibition of both innate immunity and CMI to BTV2. The sum of responses subsequent to the early IL-4 expression likely constitutes part of the unique scenario in the current BTV2-Cow experimental combination biased toward Th2 response.


Asunto(s)
Lengua Azul/inmunología , Citocinas/inmunología , Hipersensibilidad Inmediata/veterinaria , Inmunidad Innata , Leucocitos Mononucleares/virología , Animales , Virus de la Lengua Azul/clasificación , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/virología , Medios de Cultivo , Hipersensibilidad Inmediata/inmunología , Inmunidad Celular , Leucocitos Mononucleares/inmunología , Células TH1/inmunología , Células Th2/inmunología , Replicación Viral
9.
Pathogens ; 9(4)2020 Apr 03.
Artículo en Inglés | MEDLINE | ID: mdl-32260208

RESUMEN

In Taiwan, the prevalent CSFV population has shifted from the historical genotype 3.4 (94.4 strain) to the newly invading genotype 2.1 (TD/96 strain) since 1996. This study analyzed the competition between these two virus genotypes in dual infection pigs with equal and different virus populations and with maternally derived neutralizing antibodies induced by a third genotype of modified live vaccine (MLV), to simulate that occurring in natural situations in the field. Experimentally, under various dual infection conditions, with or without the presence of maternal antibodies, with various specimens from blood, oral and fecal swabs, and internal organs at various time points, the TD/96 had consistently 1.51-3.08 log higher loads than those of 94.4. A second passage of competition in the same animals further widened the lead of TD/96 as indicated by viral loads. The maternally derived antibodies provided partial protection to both wild type CSFVs and was correlated with lower clinical scores, febrile reaction, and animal mortality. In the presence of maternal antibodies, pigs could be infected by both wild type CSFVs, with TD/96 dominating. These findings partially explain the CSFV shift observed, furthering our understanding of CSFV pathogenesis in the field, and are helpful for the control of CSF.

10.
Vet Pathol ; 55(5): 673-677, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-29661121

RESUMEN

Plasmacytoid and rhabdoid variants of urothelial carcinomas (UCs) of the urinary bladder have been described in humans with plasma cell-like or rhabdoid cellular appearance and aggressive clinical outcome. Canine UC of the bladder is generally classified as papillary/nonpapillary and infiltrating/noninfiltrating with limited information regarding other histological patterns. We report 3 cases of UC of the urinary bladder showing a unique discohesive cellular morphology with malignant behavior resembling the human plasmacytoid and rhabdoid variants of UC, which may raise some difficulties in diagnosis. Epithelial-mesenchymal transition and reduced E-cadherin expression were revealed by immunohistochemistry in 2 cases, possibly explaining the discohesive and invasive behavior of the tumor cells. The findings broaden the morphological spectrum as well as the distinct clinical features of canine UC of the urinary bladder.


Asunto(s)
Carcinoma/veterinaria , Enfermedades de los Perros/patología , Transición Epitelial-Mesenquimal , Neoplasias de la Vejiga Urinaria/veterinaria , Animales , Cadherinas/metabolismo , Carcinoma/diagnóstico , Carcinoma/patología , Enfermedades de los Perros/diagnóstico , Perros , Femenino , Masculino , Vejiga Urinaria/citología , Vejiga Urinaria/patología , Neoplasias de la Vejiga Urinaria/diagnóstico , Neoplasias de la Vejiga Urinaria/patología , Urotelio/patología
11.
J Vet Diagn Invest ; 30(3): 430-437, 2018 May.
Artículo en Inglés | MEDLINE | ID: mdl-29528810

RESUMEN

Nephroblastomas are uncommon embryonal tumors in dogs. We report herein a blastema-predominant nephroblastoma with gingival metastasis in an 8-y-old Miniature Pinscher dog. Histologically, the mass was composed mainly of blastemal elements with minor epithelial and mesenchymal differentiation. Metastatic masses in the gingiva had histologic and immunohistochemical features similar to those of the primary renal nephroblastoma. Neoplastic cells were extensively positive for both vimentin and PAX8, and scattered positive for cytokeratin. Using the clinical staging of human Wilms tumor, we staged our case as stage IV with <4 mo of survival time. We summarized previous studies of canine renal and spinal nephroblastomas, and analyzed the correlations among clinical staging, histologic classification, and mean survival time of dogs with renal nephroblastomas. Clinical staging was significantly correlated with survival time, as shown in humans. In dogs, however, additional factors can potentially influence the outcome of treatment and disease development.


Asunto(s)
Enfermedades de los Perros/diagnóstico , Neoplasias Gingivales/veterinaria , Neoplasias Renales/veterinaria , Tumor de Wilms/veterinaria , Animales , Diagnóstico Diferencial , Enfermedades de los Perros/patología , Perros , Neoplasias Gingivales/diagnóstico , Neoplasias Gingivales/secundario , Neoplasias Renales/diagnóstico , Neoplasias Renales/patología , Masculino , Metástasis de la Neoplasia , Estadificación de Neoplasias/veterinaria , Tumor de Wilms/diagnóstico , Tumor de Wilms/secundario
12.
Dis Aquat Organ ; 123(3): 239-249, 2017 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-28322210

RESUMEN

A mass mortality event of captive Hong Kong warty newts Paramesotriton hongkongensis with non-granulomatous necrotic lesions occurred in Taipei Zoo, Taiwan, in 2014. Clinically, the sick newts were lethargic and often covered with water mold Saprolegnia sp. on the skin of the body trunk or extremities. Predominant pathological findings were multifocal non-granulomatous necrotic lesions in the liver, spleen, and kidneys and severe skin infection with Saprolegnia sp., with deep invasion and involvement of underlying muscles. The possibility of ranavirus infection was ruled out by negative PCR results. Unexpectedly, abundant intralesional acid-fast positive bacilli were found in the necrotic lesions of the liver, spleen, and kidney in all 14 sick newts. PCR targeting the hsp65, ITS region, and partial 16S rRNA genes was performed, and the sequence identity from amplified amplicons of hsp65 and partial 16S rRNA genes was 100% identical to that of the corresponding gene fragment of Mycobacterium marinum. Further molecular investigations demonstrated that the current M. marinum was a mycolactone-producing mycobacterium with the presence of esxA/esxB genes. Mycolactone is a plasmid-encoded, immunosuppressive, and cytotoxic toxin. The possible immunosuppression phenomenon characterized by systemic non-granulomatous necrotic lesions caused by M. marinum and the unusual deep invasive infection caused by water mold might be associated with the immunosuppressive effect of mycolactone. Therefore, it should be noted that non-granulomatous necrotic lesions in amphibians can be caused not only by ranavirus infection but also by mycobacteriosis.


Asunto(s)
Macrólidos/metabolismo , Infecciones por Mycobacterium no Tuberculosas/veterinaria , Mycobacterium marinum/metabolismo , Salamandridae/microbiología , Animales , Secuencia de Bases , ADN Bacteriano/genética , Infecciones por Mycobacterium no Tuberculosas/inmunología , Infecciones por Mycobacterium no Tuberculosas/microbiología , Infecciones por Mycobacterium no Tuberculosas/mortalidad , Mycobacterium marinum/genética , Salamandridae/inmunología
13.
Virus Res ; 228: 39-45, 2017 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-27889614

RESUMEN

Classical swine fever (CSF), an economically important and highly contagious disease of pigs, is caused by classical swine fever virus (CSFV). In Taiwan, CSFVs from field outbreaks belong to two distinct genotypes. The historical genotype 3.4 dominated from the 1920s to 1996, and since 1996, the newly invading genotype 2.1 has dominated. To explain the phenomenon of this virus shift in the field, representative viruses belonging to genotypes 2.1 and 3.4 were either inoculated alone (single infection) or co-inoculated (co-infection), both in vivo and in vitro, to compare the virus replication and pathogenesis. In pigs co-infected with the genotype 2.1 TD/96/TWN strain and the genotype 3.4 94.4/IL/94/TWN strain, the newly invading genotype 2.1 was detected earlier in the blood, oral fluid, and feces, and the viral loads were consistently and significantly higher than that of the historical genotype 3.4. In cell cultures, the ratio of secreted virus to cell-associated virus of the genotype 2.1 strain was higher than that of the genotype 3.4 strain. This study is the first to demonstrate a possible explanation of virus shift in the field, wherein the newly invading genotype 2.1 replicates more efficiently than did genotype 3.4 and outcompetes the replication and pathogenicity of genotype 3.4 in pigs in the field.


Asunto(s)
Virus de la Fiebre Porcina Clásica/fisiología , Peste Porcina Clásica/virología , Replicación Viral , Animales , Línea Celular , Peste Porcina Clásica/diagnóstico , Virus de la Fiebre Porcina Clásica/patogenicidad , Variación Genética , Genotipo , Cinética , Porcinos , Evaluación de Síntomas , Carga Viral , Acoplamiento Viral
14.
J Vet Diagn Invest ; 28(6): 744-749, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27698165

RESUMEN

An ~21-year-old female Barbary sheep (Ammotragus lervia) died spontaneously following a lengthy episode of difficulty in walking. An ~6 × 3 × 3 cm, unilocular cystic growth was found in the cranioventral thorax. The fibrotic cystic wall, lined by a single layer of flattened to cuboidal epithelial cells, was invaginated and partially encircled solid masses of fusiform neoplastic cells with multiple intratumoral cystic structures. The fusiform neoplastic cells were intensely positive for cytokeratin (CK) and partially positive for α-smooth muscle actin and vimentin, but negative for thyroid transcription factor-1 (TTF-1) and neuron-specific enolase (NSE). The intratumoral cysts were lined by CK-positive but TTF-1- negative, NSE-negative, flattened, cuboidal to columnar epithelial cells, suggestive of cystically dilated medullary duct epithelium-derived structures. Based on the location and histopathologic findings of the growth, concurrent spindle-cell thymoma and thymic cysts was diagnosed. We also discuss the correlation between thymic cysts and thymoma and review the literature of thymomas in ovine and wildlife species.


Asunto(s)
Quiste Mediastínico/veterinaria , Rumiantes , Timoma/veterinaria , Neoplasias del Timo/veterinaria , Animales , Diagnóstico Diferencial , Femenino , Quiste Mediastínico/complicaciones , Quiste Mediastínico/diagnóstico , Timoma/complicaciones , Timoma/diagnóstico , Neoplasias del Timo/complicaciones , Neoplasias del Timo/diagnóstico
15.
J Vet Diagn Invest ; 28(5): 599-603, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27493139

RESUMEN

A 5-year-old male captive corn snake (Pantherophis guttatus) with caudal coelomic swelling was admitted for surgical treatment. Laparotomy revealed a 5 × 4 × 2.5 cm, firm, expansile, irregularly shaped mass arising from the middle portion of the right kidney with a mild lobulated pattern and mottled white-to-tan. Microscopically, the mass was composed of numerous bizarre angulated tubules of polygonal neoplastic cells separated by a scirrhous stroma with remarkable heterophilic infiltrates. The neoplastic cells were nonciliated and mucin secreting, with abundant brightly eosinophilic cytoplasm. There were marked cellular and nuclear atypia, frequent cell individualization, and stromal invasion, indicative of malignant behavior, which was confirmed by metastasis to the left kidney 1.5 months postoperatively. Both neoplastic epithelial cells and mesenchymal cells contributing to the scirrhous stroma had variable immunopositivity for pan-cytokeratin. The neoplasm was considered a renal adenocarcinoma resembling human collecting duct carcinoma.


Asunto(s)
Carcinoma de Células Renales/veterinaria , Neoplasias Renales/veterinaria , Serpientes , Animales , Carcinoma de Células Renales/diagnóstico , Carcinoma de Células Renales/patología , Diagnóstico Diferencial , Neoplasias Renales/diagnóstico , Neoplasias Renales/patología , Masculino
16.
Vet Microbiol ; 182: 150-5, 2016 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-26711042

RESUMEN

Porcine teschoviruses (PTVs) belong to the genus Teschovirus within the family Picornaviridae. PTVs are universal contaminants in pig herds in endemic and multi-infection statuses. Previous research has demonstrated PTV antigens and nucleic acid in renal glomeruli and tubular epithelia, suggesting the possibility that PTVs might be shed and transmitted via urine. The study aimed to demonstrate, in the context of pathogenesis, the presence of PTVs in the urine of naturally infected pigs. Viral loads of fluid and tissue samples quantified by an established qRT-PCR showed detection rates of 100% by head and in urine, feces, plasma and nasal swabs, and 38% in kidney. As predicted, PTVs were present in urine at 10(4.02 ± 1.45) copies/100 µl volume, equivalent to 17% of that in plasma. No significant differences were observed between healthy and culled pigs or among the 7 sampled herds. The presence of PTVs in urine was further substantiated by molecular serotyping. In particular, PTV-10 was identified in the urine of 3 piglets from 3 separate herds, consistent with the most prevalent serotype found in this study, and in plasma. The urine mixes with feces to form slurry making it easier for PTV to spread and contaminate the environment.


Asunto(s)
Enfermedades Endémicas/veterinaria , Infecciones por Picornaviridae/veterinaria , Enfermedades de los Porcinos/virología , Teschovirus/fisiología , Orina/virología , Animales , Infecciones por Picornaviridae/genética , Infecciones por Picornaviridae/transmisión , Infecciones por Picornaviridae/virología , Serogrupo , Sus scrofa/virología , Porcinos , Enfermedades de los Porcinos/genética , Enfermedades de los Porcinos/transmisión , Teschovirus/genética , Teschovirus/aislamiento & purificación , Carga Viral , Esparcimiento de Virus
17.
Arch Virol ; 160(11): 2709-18, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26246243

RESUMEN

Porcine reproductive and respiratory syndrome virus (PRRSV) was first identified in Taiwan in 1991, but the genetic diversity and evolution of PRRSV has not been thoroughly investigated over the past 20 years. The aim of this study was to bridge the gap in understanding of its molecular epidemiology. A total of 31 PRRSV strains were collected and sequenced. The sequences were aligned using the MUSCLE program, and phylogenetic analysis were performed by the maximum-likelihood method and the neighbor-joining method using MEGA 5.2 software. In the early 1990s, two prototype strains, WSV and MD001 of the North American genotype, were first identified. Over the years, both viruses evolved separately. The population dynamics of PRRSV revealed that the strains of the MD001 group were predominant in Taiwan. Evolution was manifested in changes in the nsp2 and ORF5 genes. In addition, a suspected newly invading exotic strain was recovered in 2013, suggesting that international spread is still taking place and that it is affecting the population dynamics. Overall, the results provide an important basis for vaccine development for the control and prevention of PRRS.


Asunto(s)
Síndrome Respiratorio y de la Reproducción Porcina/virología , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Virus del Síndrome Respiratorio y Reproductivo Porcino/aislamiento & purificación , Animales , Variación Genética , Genoma Viral , Genotipo , Epidemiología Molecular , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Filogenia , Síndrome Respiratorio y de la Reproducción Porcina/epidemiología , Virus del Síndrome Respiratorio y Reproductivo Porcino/química , Virus del Síndrome Respiratorio y Reproductivo Porcino/clasificación , Alineación de Secuencia , Porcinos , Taiwán/epidemiología , Proteínas Virales/química , Proteínas Virales/genética
18.
Viruses ; 7(7): 3506-29, 2015 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-26131960

RESUMEN

Pestiviruses, which include economically important animal pathogens such as bovine viral diarrhea virus and classical swine fever virus, possess three envelope glycoproteins, namely Erns, E1, and E2. This article discusses the structures and functions of these glycoproteins and their effects on viral pathogenicity in cells in culture and in animal hosts. E2 is the most important structural protein as it interacts with cell surface receptors that determine cell tropism and induces neutralizing antibody and cytotoxic T-lymphocyte responses. All three glycoproteins are involved in virus attachment and entry into target cells. E1-E2 heterodimers are essential for viral entry and infectivity. Erns is unique because it possesses intrinsic ribonuclease (RNase) activity that can inhibit the production of type I interferons and assist in the development of persistent infections. These glycoproteins are localized to the virion surface; however, variations in amino acids and antigenic structures, disulfide bond formation, glycosylation, and RNase activity can ultimately affect the virulence of pestiviruses in animals. Along with mutations that are driven by selection pressure, antigenic differences in glycoproteins influence the efficacy of vaccines and determine the appropriateness of the vaccines that are currently being used in the field.


Asunto(s)
Enfermedades de los Bovinos/virología , Infecciones por Pestivirus/veterinaria , Pestivirus/metabolismo , Enfermedades de los Porcinos/virología , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/metabolismo , Animales , Bovinos , Pestivirus/química , Pestivirus/genética , Pestivirus/patogenicidad , Infecciones por Pestivirus/virología , Porcinos , Proteínas del Envoltorio Viral/genética , Internalización del Virus
19.
J Virol Methods ; 201: 13-9, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24560782

RESUMEN

The porcine respiratory disease complex (PRDC) is the most common disease in commercial pork production worldwide. Porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), the most important agents of PRDC, usually co-infect in the same pigs. In order to survey the prevalence of PCV2 and PRRSV in pigs of various ages, a duplex reverse transcription real-time PCR (DRT-rPCR) was developed and applied in the present study. The DRT-rPCR did not cross-react with 10 swine viruses other than PCV2 and PRRSV, with detection limits of 1 TCID50/ml for PCV2 and 6.3 TCID50/ml for PRRSV. Surveillance using DRT-rPCR together with serology revealed that in the five farms studied, pigs were most susceptible to PRRSV at 6-14 weeks of age, whereas susceptibility to PCV2 varied by the management system but was mostly at 10-14 weeks of age. Cross analysis of viral loads versus antibody titers revealed that PCV2 load was affected negatively by anti-PCV2 ORF2 antibody, which constituted the most important non-infectious factor affecting the development of PMWS. These results indicated that DRT-rPCR was developed and applied successfully to the surveillance of PCV2 and PRRSV in the field.


Asunto(s)
Infecciones por Circoviridae/veterinaria , Circovirus/aislamiento & purificación , Reacción en Cadena de la Polimerasa Multiplex/métodos , Síndrome Respiratorio y de la Reproducción Porcina/diagnóstico , Virus del Síndrome Respiratorio y Reproductivo Porcino/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Animales , Anticuerpos Antivirales/sangre , Infecciones por Circoviridae/diagnóstico , Infecciones por Circoviridae/virología , Circovirus/genética , Monitoreo Epidemiológico , Síndrome Respiratorio y de la Reproducción Porcina/virología , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Sensibilidad y Especificidad , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/virología , Carga Viral
20.
Vet Microbiol ; 168(1): 69-77, 2014 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-24268804

RESUMEN

Porcine teschoviruses (PTVs) belong to the genus Teschovirus within the family Picornaviridae. PTVs are universal contaminants in pig herds in endemic and multi-infection status. To further the understanding of PTV pathogenesis in endemically infected pigs, a set of samples was studied by real time reverse transcription PCR (qRT-PCR) to quantitate viral loads in tissues and by in situ hybridization (ISH) to locate PTV signals in target cells, both targeting the 5'-NTR. cRNA of PTV-1 and PTV-7, in vitro transcribed from cloned fragments of 5'-NTR of 2 viruses, was used to construct standard curves and to run parallel in qRT-PCR, which had detection limits of 10(1) copies/per reaction, with a linearity in between 10(1) and 10(7) copies/per reaction and correlation coefficients of 0.997-0.9988. The qRT-PCR specifically amplified RNA from PTV-1 to -11, while excluding those of Sapelovirus, PEV-9 and PEV-10. Inguinal lymph node (LN) had the highest viral load of all (assuming 100%), followed by ileac LN (89-91%), tonsil (66-68%), ileum (59-60%), spleen (38-40%), and kidney (30-31%), with the least in brain (22.9%) of the inguinal LN. The 22.9% load in brain was higher than that anticipated from a simple fecal-oral-viremia operative model. The results suggested in addition that intranasal infection and retrograding axonal infection from the tonsils were equally operative and significant. ISH revealed PTV signals in a wider variety of tissue cell types than before. PTV signals were noted most impressively in neurons of the cerebral cortex and hippocampus and in the dark zone of the germinal center and adjacent paracortex of regional LN. Multiple operative models indicated that PTVs seemed to have no difficulty invading the brain. The key to whether encephalitis would ensue resided in the animal's immune status and topographic differences of neurons' susceptibilities to PTVs. When common co-infected agents are present, as is typical in the field, PTVs may synergize in causing diseases.


Asunto(s)
Enfermedades Endémicas/veterinaria , Infecciones por Picornaviridae/veterinaria , Enfermedades de los Porcinos/patología , Enfermedades de los Porcinos/virología , Teschovirus/patogenicidad , Animales , Heces/virología , Infecciones por Picornaviridae/patología , Infecciones por Picornaviridae/virología , Reproducibilidad de los Resultados , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Sensibilidad y Especificidad , Porcinos , Carga Viral
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