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1.
Sheng Wu Gong Cheng Xue Bao ; 37(9): 3221-3230, 2021 Sep 25.
Artículo en Chino | MEDLINE | ID: mdl-34622630

RESUMEN

The aim of this study was to develop a blocking enzyme-linked immunosorbent assay (bELISA) based on a biotinylated nanobody target the S1 protein of porcine epidemic diarrhea virus (PEDV) for detecting the anti-PEDV antibodies and evaluating the immune effect of the vaccine. The gene encoding the single-domain antibody sdAb3 target the PEDV S1 protein was amplified and the Avitag sequence was fused at its 3'-end. The PCR product was cloned into the expression vector pET-21b for expression and purification of the sdAb3-Avitag protein. The purified sdAb3-Avitag fusion protein was biotinylated and its activity was determined. Using the recombinant S1 protein as a coating antigen, a bELISA was established and optimized. Serum samples were tested in parallel by the bELISA and a commercial kit. The recombinant vector pET21b-sdAb3-Avitag was constructed to express the tagged sdAb3. After induction for expression, the biotin-labeled sdAb3 (sdAb3-Biotin) with high purity and good activity was obtained. For the optimized bELISA, the coating concentration of the S1 protein was 200 ng/well, the serum dilution was 1:2 and incubated for 2 h, the dilution ratio of the biotinylated sdAb3 was 1:8 000 and incubated for 30 min, the dilution of the enzyme-labeled antibody was 1:5 000 and incubated for 30 min. The bELISA had no cross reaction with the sera of major porcine viruses including transmissible gastroenteritis virus, porcine reproductive and respiratory syndrome virus and showed good specificity and reproducibility. For a total of 54 porcine serum samples tested, the overall compliance rate of the bELISA with a commercial kit was 92.56%. This study developed a rapid and reliable bELISA method, which can be used for serosurveillance and vaccine evaluation for PEDV.


Asunto(s)
Infecciones por Coronavirus , Virus de la Diarrea Epidémica Porcina , Anticuerpos de Dominio Único , Enfermedades de los Porcinos , Animales , Anticuerpos Antivirales , Infecciones por Coronavirus/veterinaria , Ensayo de Inmunoadsorción Enzimática , Virus de la Diarrea Epidémica Porcina/genética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Porcinos
2.
Sheng Wu Gong Cheng Xue Bao ; 37(8): 2603-2613, 2021 Aug 25.
Artículo en Chino | MEDLINE | ID: mdl-34472281

RESUMEN

Porcine epidemic diarrhea (PED) is a major disease of pigs that inflicts heavy losses on the global pig industry. The etiologic agent is the porcine epidemic diarrhea virus (PEDV), which is assigned to the genus Alphacoronavirus in the family Coronaviridae. This review consists of five parts, the first of which provides a brief introduction to PEDV and its epidemiology. Part two outlines the passive immunity in new born piglets and the important role of colostrum, while the third part summarizes the characteristics of the immune systems of pregnant sows, discusses the concept of the "gut-mammary gland-secretory IgA(sIgA) axis" and the possible underpinning mechanisms, and proposes issues to be addressed when designing a PEDV live vaccine. The final two parts summarizes the advances in the R&D of PEDV vaccines and prospects future perspectives on prevention and control of PEDV, respectively.


Asunto(s)
Infecciones por Coronavirus , Virus de la Diarrea Epidémica Porcina , Enfermedades de los Porcinos , Vacunas Virales , Animales , Anticuerpos Antivirales , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/veterinaria , Femenino , Inmunización , Embarazo , Porcinos , Enfermedades de los Porcinos/prevención & control
3.
Sheng Wu Gong Cheng Xue Bao ; 37(8): 2779-2785, 2021 Aug 25.
Artículo en Chino | MEDLINE | ID: mdl-34472295

RESUMEN

To investigate whether the engineered Lactobacillus plantarum expressing the porcine epidemic diarrhea virus (PEDV) S1 gene can protect animals against PEDV, guinea pigs were fed with recombinant L. plantarum containing plasmid PVE5523-S1, with a dose of 2×108 CFU/piece, three times a day, at 14 days intervals. Guinea pigs fed with wild type L. plantarum and the engineered L. plantarum containing empty plasmid pVE5523 were used as negative controls. For positive control, another group of guinea pigs were injected with live vaccine for porcine epidemic diarrhea and porcine infectious gastroenteritis (HB08+ZJ08) by intramuscular injection, with a dose of 0.2 mL/piece, three times a day, at 14 days intervals. Blood samples were collected from the hearts of the four groups of guinea pigs at 0 d, 7 d, 14 d, 24 d, 31 d, 41 d and 48 d, respectively, and serum samples were isolated for antibody detection and neutralization test analysis by enzyme-linked immunosorbent assay (ELISA). The spleens of guinea pigs were also aseptically collected to perform spleen cells proliferation assay. The results showed that the engineered bacteria could stimulate the production of secretory antibody sIgA and specific neutralizing antibody, and stimulate the increase of IL-4 and IFN-γ, as well as the proliferation of spleen cells. These results indicated that the engineered L. plantarum containing PEDV S1 induced specific immunity toward PEDV in guinea pigs, which laid a foundation for subsequent oral vaccine development.


Asunto(s)
Infecciones por Coronavirus , Lactobacillus plantarum , Virus de la Diarrea Epidémica Porcina , Enfermedades de los Porcinos , Vacunas Virales , Animales , Anticuerpos Antivirales , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/veterinaria , Cobayas , Lactobacillus plantarum/genética , Virus de la Diarrea Epidémica Porcina/genética , Porcinos , Vacunas Virales/genética
4.
Avian Dis ; 65(1): 18-25, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-34339117

RESUMEN

Severity of the tracheal histologic inflammatory response induced in broilers by ocular inoculation of two infectious bronchitis (IBV) and three Newcastle disease virus (NDV) commercial vaccines were evaluated. The vaccine was delivered by eye drop with a coarse spray to day-old chicks. The vaccines were given individually or in various combinations and were evaluated relative to nonvaccinated controls. Evaluations were performed on postvaccination (PV) days 7 and 14. Histologic endpoints included semiquantitative severity scoring of inflammatory components and quantitative morphometric determinations of inflammatory cell concentration, mucosal thickness, and percentage of ciliated mucosal surface. Strong positive correlations were observed between routine severity scoring and morphometric inflammatory parameters, whereas a negative correlation was present between inflammation severity and the percentage of mucosal ciliation. Variable, sometimes extensive, and often statistically significant differences in inflammatory responses were observed between the various vaccines. One IBV Massachusetts strain vaccine (IBV-A) produced the greatest overall inflammatory response when given alone or in combination with the NDV vaccines. Enhancement of tracheitis was seen on PV day 14 by covaccination of IBV-A with the NDV vaccines, but not by covaccination of another IBV Massachusetts strain vaccine (IBV-B) with NDV. Reduction in cilia percentage was observed for all vaccine groups relative to controls on PV day 7. However, although reactive cilia regeneration occurred on PV day 14 for most vaccine groups, a cilia regenerative response was not apparent for individual or NDV combination vaccination for IBV-A. The study also demonstrates that substantial microscopic trachea pathology may be present in vaccinated birds not exhibiting apparent clinical respiratory signs.


Asunto(s)
Pollos , Infecciones por Coronavirus/veterinaria , Virus de la Bronquitis Infecciosa/inmunología , Enfermedad de Newcastle/prevención & control , Virus de la Enfermedad de Newcastle/inmunología , Enfermedades de las Aves de Corral/prevención & control , Vacunación/veterinaria , Vacunas Virales/efectos adversos , Animales , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/virología , Enfermedad de Newcastle/virología , Enfermedades de las Aves de Corral/virología , Tráquea/patología , Tráquea/virología , Vacunación/efectos adversos , Vacunas Combinadas/efectos adversos
5.
Avian Dis ; 65(1): 40-45, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-34339120

RESUMEN

We performed viral metagenomics analysis of Japanese quail affected with enteritis to elucidate the viral etiology. Metagenomics generated 21,066,442 sequence reads via high-throughput sequencing, with a mean length of 136 nt. Enrichment in viral sequences suggested that at least three viruses were present in quail samples. Coronavirus and picornavirus were identified and are known as pathogens causing quail enteritis that match the observed morphology. Abundant reads of coronavirus from quail samples yielded four fragment sequences exhibiting six genomes of avian coronavirus. Sequence analysis showed that this quail coronavirus was related to turkey coronavirus and chicken infectious bronchitis virus. Quail picornavirus 8177 bp in size was identified and was similar to the QPV1/HUN/01 virus detected in quails without clinical symptoms in Hungary with 84.6% nucleotide and 94.6% amino acid identity. Our results are useful for understanding the genetic diversity of quail viruses. Further studies must be performed to determine whether quail coronavirus and quail picornavirus are pathogens of the digestive tract of quails.


Asunto(s)
Infecciones por Coronavirus/veterinaria , Coronavirus/genética , Coturnix/virología , Enteritis/veterinaria , Metagenómica/métodos , Enfermedades de las Aves de Corral/virología , Animales , Coronavirus/aislamiento & purificación , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/virología , Enteritis/epidemiología , Enteritis/virología , Genoma Viral , Picornaviridae/aislamiento & purificación , Infecciones por Picornaviridae/epidemiología , Infecciones por Picornaviridae/veterinaria , Infecciones por Picornaviridae/virología , Enfermedades de las Aves de Corral/epidemiología , República de Corea/epidemiología
6.
Avian Dis ; 65(1): 188-197, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-34339139

RESUMEN

Infectious bronchitis virus (IBV) causes significant losses in the poultry industry throughout the world. Here we characterize the lesions of infectious bronchitis (IB) and IBV prevalence and identify the circulating strains in small flocks in California. Backyard chickens (BYCs) submitted to the Davis (Northern California; NorCal) and San Bernardino (Southern California; SoCal) branches of the California Animal Health and Food Safety Laboratory System from January through March 2019 were included in the study. Trachea, kidney, and cecal tonsils were collected for real-time reverse transcriptase (qRT)-PCR, histology, immunohistochemistry (IHC), and sequence analysis. A total of 50 chickens out of 169 submissions tested positive for IBV by qRT-PCR. Of these, 16% (20/123) were from NorCal and 65% (30/46) from SoCal laboratory. The cecal tonsil was the most frequently positive tissue by qRT-PCR and IHC. Lymphoplasmacytic tracheitis was the most frequent histopathologic finding in 24 of 39 birds, while the kidney showed interstitial nephritis, tubular necrosis, tubular dilation, and/or gout in 14 of 43 chickens. Infectious bronchitis virus played a primary role or a synergistic effect in the mortality of chickens that succumbed to other infectious diseases. The sequences of IBV detected in 22 birds were analyzed, and 14 strains were most similar to CA1737. One strain each matched Conn46, Cal99, and ArkDPI, and the remaining five did not have a substantial match to any available reference strains. The findings in this study indicate that small flocks can be reservoirs of IBV and might facilitate evolution of new variants as well as reversion of attenuated strains to virulence.


Asunto(s)
Pollos , Infecciones por Coronavirus/veterinaria , Virus de la Bronquitis Infecciosa/aislamiento & purificación , Enfermedades de las Aves de Corral/virología , Animales , California/epidemiología , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/virología , Virus de la Bronquitis Infecciosa/clasificación , Enfermedades de las Aves de Corral/epidemiología
7.
Poult Sci ; 100(9): 101324, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34358949

RESUMEN

Avian infectious bronchitis (IB), a highly contagious disease hazardous to the poultry industry, is caused by an etiological agent called the infectious bronchitis virus (IBV). Some IBV strains (IBVs) alone usually do not cause high mortality in field conditions if not with secondary pathogens including Escherichia coli (E. coli). Herein, we established an IBV and E. coli co-infection model to evaluate the protective efficacy of two IBV vaccine strains against a new emerging genotype GVI-1 with mild virulence in experimental conditions. Chickens were inoculated with IBV field isolate ZQX (genotype GVI-1) and challenged 4 dlater with the E. coli strain MS160427 (serotype O8). Subsequently, these chickens were euthanized at seven days postchallenge (d.p.c.) with E. coli. An autopsy revealed that lesions in the IBV plus E. coli co-infection group were more severe than those in the IBV-infected group. This pathological model was used to assess the protective effect of two commonly used vaccine strains (H120 and 4/91) against the IBV ZQX strain, and a significantly better protective efficacy was observed for 4/91 compared with H120. Thus, IBV and E. coli co-infection could be employed in assessing the protective efficacy of IBV vaccines.


Asunto(s)
Bronquitis , Coinfección , Infecciones por Coronavirus , Virus de la Bronquitis Infecciosa , Enfermedades de las Aves de Corral , Vacunas Virales , Animales , Bronquitis/veterinaria , Pollos , Coinfección/veterinaria , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/veterinaria , Escherichia coli , Enfermedades de las Aves de Corral/prevención & control
8.
Viruses ; 13(8)2021 07 29.
Artículo en Inglés | MEDLINE | ID: mdl-34452362

RESUMEN

The revealed prevalence of coronaviruses in wild bird populations in Poland was 4.15% and the main reservoirs were birds from orders Anseriformes and Charadriiformes, with a prevalence of 3.51% and 5.59%, respectively. Gammacoronaviruses were detected more often than deltacoronaviruses, with detection rates of 3.5% and 0.7%, respectively. Gammacoronaviruses were detected in birds belonging to six orders, including Anseriformes, Charadriiformes, Columbiformes, Galliformes, Gruiformes, and Passeriformes, indicating a relatively wide host range. Interestingly, this was the only coronavirus detected in Anseriformes (3.51%), while in Charadriiformes, the prevalence was 3.1%. The identified gammacoronaviruses belonged to the Igacovirus and Brangacovirus subgeneras. Most of these were igacoviruses and formed a common phylogenetic group with a Duck Coronavirus 2714 and two with an Avian Coronavirus/Avian Coronavirus9203, while the viruses from the pigeons formed a distinct "pigeon-like" group, not yet officially represented. The presence of deltacoronaviruses was detected in birds belonging to three orders, Charadriiformes, Galliformes, and Suliformes indicating a narrower host range. Most identified deltacoronaviruses belonged to the Buldecovirus subgenus, while only one belonged to Herdecovirus. Interestingly, the majority of buldecoviruses were identified in gulls, and they formed a distinct phylogenetic lineage not represented by any officially ratified virus species. Another separate group of buldecoviruses, also not represented by the official species, was formed by a virus identified in a common snipe. Only one identified buldecovirus (from common pheasant) formed a group with the ratified species Coronavirus HKU15. The results obtained indicate the high diversity of detected coronaviruses, and thus also the need to update their taxonomy (establishing new representative virus species). The serological studies performed revealed antibodies against an infectious bronchitis virus in the sera of white storks and mallards.


Asunto(s)
Animales Salvajes/virología , Biodiversidad , Enfermedades de las Aves/virología , Infecciones por Coronavirus/veterinaria , Gammacoronavirus/aislamiento & purificación , Animales , Animales Salvajes/clasificación , Anseriformes/virología , Charadriiformes/virología , Columbiformes/virología , Infecciones por Coronavirus/virología , Patos/virología , Galliformes/virología , Gammacoronavirus/clasificación , Gammacoronavirus/genética , Filogenia , Polonia
9.
Viruses ; 13(8)2021 07 30.
Artículo en Inglés | MEDLINE | ID: mdl-34452372

RESUMEN

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel coronavirus in humans, has expanded globally over the past year. COVID-19 remains an important subject of intensive research owing to its huge impact on economic and public health globally. Based on historical archives, the first coronavirus-related disease recorded was possibly animal-related, a case of feline infectious peritonitis described as early as 1912. Despite over a century of documented coronaviruses in animals, the global animal industry still suffers from outbreaks. Knowledge and experience handling animal coronaviruses provide a valuable tool to complement our understanding of the ongoing COVID-19 pandemic. In this review, we present an overview of coronaviruses, clinical signs, COVID-19 in animals, genome organization and recombination, immunopathogenesis, transmission, viral shedding, diagnosis, treatment, and prevention. By drawing parallels between COVID-19 in animals and humans, we provide perspectives on the pathophysiological mechanisms by which coronaviruses cause diseases in both animals and humans, providing a critical basis for the development of effective vaccines and therapeutics against these deadly viruses.


Asunto(s)
Enfermedades de los Animales/virología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Coronavirus/fisiología , Enfermedades de los Animales/epidemiología , Animales , COVID-19/epidemiología , COVID-19/virología , Coronavirus/genética , Infecciones por Coronavirus/epidemiología , Humanos , Salud Pública , SARS-CoV-2/genética , SARS-CoV-2/fisiología
10.
Emerg Infect Dis ; 27(9): 2301-2311, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34423762

RESUMEN

After the first detection of Middle East respiratory syndrome coronavirus (MERS-CoV) in camels in Jordan in 2013, we conducted 2 consecutive surveys in 2014-2015 and 2017-2018 investigating risk factors for MERS-CoV infection among camel populations in southern Jordan. Multivariate analysis to control for confounding demonstrated that borrowing of camels, particularly males, for breeding purposes was associated with increased MERS-CoV seroprevalence among receiving herds, suggesting a potential route of viral transmission between herds. Increasing age, herd size, and use of water troughs within herds were also associated with increased seroprevalence. Closed herd management practices were found to be protective. Future vaccination strategies among camel populations in Jordan could potentially prioritize breeding males, which are likely to be shared between herds. In addition, targeted management interventions with the potential to reduce transmission between herds should be considered; voluntary closed herd schemes offer a possible route to achieving disease-free herds.


Asunto(s)
Infecciones por Coronavirus , Coronavirus del Síndrome Respiratorio de Oriente Medio , Animales , Camelus , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/veterinaria , Jordania/epidemiología , Masculino , Factores de Riesgo , Estudios Seroepidemiológicos
11.
J Gen Virol ; 102(8)2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-34424155

RESUMEN

Infectious bronchitis virus (IBV) is an economically important coronavirus, causing damaging losses to the poultry industry worldwide as the causative agent of infectious bronchitis. The coronavirus spike (S) glycoprotein is a large type I membrane protein protruding from the surface of the virion, which facilitates attachment and entry into host cells. The IBV S protein is cleaved into two subunits, S1 and S2, the latter of which has been identified as a determinant of cellular tropism. Recent studies expressing coronavirus S proteins in mammalian and insect cells have identified a high level of glycosylation on the protein's surface. Here we used IBV propagated in embryonated hens' eggs to explore the glycan profile of viruses derived from infection in cells of the natural host, chickens. We identified multiple glycan types on the surface of the protein and found a strain-specific dependence on complex glycans for recognition of the S2 subunit by a monoclonal antibody in vitro, with no effect on viral replication following the chemical inhibition of complex glycosylation. Virus neutralization by monoclonal or polyclonal antibodies was not affected. Following analysis of predicted glycosylation sites for the S protein of four IBV strains, we confirmed glycosylation at 18 sites by mass spectrometry for the pathogenic laboratory strain M41-CK. Further characterization revealed heterogeneity among the glycans present at six of these sites, indicating a difference in the glycan profile of individual S proteins on the IBV virion. These results demonstrate a non-specific role for complex glycans in IBV replication, with an indication of an involvement in antibody recognition but not neutralisation.


Asunto(s)
Coronavirus/fisiología , Polisacáridos/metabolismo , Glicoproteína de la Espiga del Coronavirus/química , Glicoproteína de la Espiga del Coronavirus/metabolismo , Alcaloides/química , Alcaloides/farmacología , Secuencia de Aminoácidos , Animales , Sitios de Unión , Células Cultivadas , Cromatografía Liquida , Biología Computacional/métodos , Coronavirus/efectos de los fármacos , Infecciones por Coronavirus/veterinaria , Regulación Viral de la Expresión Génica , Glicosilación/efectos de los fármacos , Virus de la Bronquitis Infecciosa/fisiología , Modelos Moleculares , Conformación Molecular , Peso Molecular , Pruebas de Neutralización , Oligosacáridos/química , Oligosacáridos/metabolismo , Polisacáridos/química , Enfermedades de las Aves de Corral/virología , Transporte de Proteínas , Espectrometría de Masa por Ionización de Electrospray , Glicoproteína de la Espiga del Coronavirus/genética , Relación Estructura-Actividad , Replicación Viral/efectos de los fármacos
12.
Avian Dis ; 65(3): 364-372, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34427409

RESUMEN

We previously reported that recombinant Newcastle disease virus LaSota (rLS) expressing infectious bronchitis virus (IBV) Arkansas (Ark)-type trimeric spike (S) ectodomain (Se; rLS/ArkSe) provides suboptimal protection against IBV challenge. We have now developed rLS expressing chicken granulocyte-macrophage colony-stimulating factor (GMCSF) and IBV Ark Se in an attempt to enhance vaccine effectiveness. In the current study, we first compared protection conferred by vaccination with rLS/ArkSe and rLS/ArkSe.GMCSF. Vaccinated chickens were challenged with virulent Ark, and protection was determined by clinical signs, viral load, and tracheal histomorphometry. Results showed that coexpression of GMCSF and the Se from rLS significantly reduced tracheal viral load and tracheal lesions compared with chickens vaccinated with rLS/ArkSe. In a second experiment, we evaluated enhancement of cross-protection of a Massachusetts (Mass) attenuated vaccine by priming or boosting with rLS/ArkSe.GMCSF. Vaccinated chickens were challenged with Ark, and protection was evaluated. Results show that priming or boosting with the recombinant virus significantly increased cross-protection conferred by Mass against Ark virulent challenge. Greater reductions of viral loads in both trachea and lachrymal fluids were observed in chickens primed with rLS/ArkSe.GMCSF and boosted with Mass. Consistently, Ark Se antibody levels measured with recombinant Ark Se protein-coated ELISA plates 14 days after boost were significantly higher in these chickens. Unexpectedly, the inverse vaccination scheme, that is, priming with Mass and boosting with the recombinant vaccine, proved somewhat less effective. We concluded that a prime and boost strategy by using rLS/ArkSe.GMCSF and the worldwide ubiquitous Mass attenuated vaccine provides enhanced cross-protection. Thus, rLS/GMCSF coexpressing the Se of regionally relevant IBV serotypes could be used in combination with live Mass to protect against regionally circulating IBV variant strains.


Asunto(s)
Infecciones por Coronavirus/veterinaria , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Virus de la Bronquitis Infecciosa/inmunología , Virus de la Enfermedad de Newcastle/genética , Enfermedades de las Aves de Corral/prevención & control , Glicoproteína de la Espiga del Coronavirus/inmunología , Animales , Anticuerpos Antivirales/inmunología , Pollos/genética , Pollos/inmunología , Pollos/virología , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/virología , Protección Cruzada , Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Virus de la Bronquitis Infecciosa/química , Virus de la Bronquitis Infecciosa/genética , Virus de la Bronquitis Infecciosa/fisiología , Virus de la Enfermedad de Newcastle/metabolismo , Enfermedades de las Aves de Corral/inmunología , Enfermedades de las Aves de Corral/virología , Dominios Proteicos , Glicoproteína de la Espiga del Coronavirus/administración & dosificación , Glicoproteína de la Espiga del Coronavirus/química , Glicoproteína de la Espiga del Coronavirus/genética , Tráquea/inmunología , Tráquea/virología , Vacunación , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunología , Carga Viral
13.
Res Vet Sci ; 139: 32-42, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34246941

RESUMEN

Porcine epidemic diarrhea virus (PEDV) is an enteropathogenic coronavirus; it causes diarrhea in pigs and is associated with high morbidity and mortality in sucking piglets. In this study, we performed in vitro and in vivo experiments to determine the inhibitory effects of Lactobacillus plantarum metabolites (LPM) on PEDV replication. Gas chromatography-mass spectrometry revealed exopolysaccharides to be one of the main components of LPM. We then determine whether L. plantarum exopolysaccharides (LPE) have an antiviral effect and also detected the expression levels of the apoptosis-related genes Bax and Bcl-2 and of the pro-apoptotic protein caspase-3. Further, we assessed the transcription levels of an immune-related protein (STAT1) and antiviral factors (MX1, MX2, ISG15, ZAP, PKR, and OAS1). Our results showed that the most effective method was to pretreat cells with LPM and that the optimal dose of LPM that could be safely administered to Vero cells was 1/8 times of the stock solution. LPE had a strong inhibitory effect on PEDV; the most effective method of administration was to co-incubate cells with LPE and PEDV, and the optimal concentration of LPE was 1.35 mg/mL. To conclude, LPE prevented PEDV adsorption and also alleviated inflammatory responses and induced early apoptosis of injured cells, but it could not regulate the immune function of cells.


Asunto(s)
Lactobacillus plantarum/metabolismo , Polisacáridos/farmacología , Polisacáridos/uso terapéutico , Virus de la Diarrea Epidémica Porcina/efectos de los fármacos , Virus de la Diarrea Epidémica Porcina/crecimiento & desarrollo , Enfermedades de los Porcinos/tratamiento farmacológico , Enfermedades de los Porcinos/virología , Replicación Viral/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Chlorocebus aethiops , Infecciones por Coronavirus/tratamiento farmacológico , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Diarrea/tratamiento farmacológico , Diarrea/veterinaria , Diarrea/virología , Inflamación/tratamiento farmacológico , Virus de la Diarrea Epidémica Porcina/inmunología , Porcinos , Enfermedades de los Porcinos/inmunología , Células Vero , Acoplamiento Viral/efectos de los fármacos
14.
Viruses ; 13(7)2021 06 25.
Artículo en Inglés | MEDLINE | ID: mdl-34201926

RESUMEN

The SARS-CoV-2 coronavirus is the focus of attention as it has caused more than three million human deaths globally. This and other coronaviruses, such as MERS-CoV, have been suggested to be related to coronaviruses that are hosted in bats. This work shows, through a bibliographic review, the frequency of detection of coronavirus in bats species of the Americas. The presence of coronavirus in bats has been examined in 25 investigations in 11 countries of the Americas between 2007 and 2020. Coronaviruses have been explored in 9371 individuals from 160 species of bats, and 187 coronavirus sequences have been deposited in GenBank distributed in 43 species of bats. While 91% of the coronaviruses sequences identified infect a single species of bat, the remainder show a change of host, dominating the intragenera change. So far, only Mex-CoV-6 is related to MERS-CoV, a coronavirus pathogenic for humans, so further coronavirus research effort in yet unexplored bat species is warranted.


Asunto(s)
Quirópteros/virología , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/veterinaria , Coronavirus/clasificación , Coronavirus/genética , Américas/epidemiología , Animales , COVID-19/epidemiología , Coronavirus/aislamiento & purificación , Infecciones por Coronavirus/virología , Humanos , Coronavirus del Síndrome Respiratorio de Oriente Medio/genética , Coronavirus del Síndrome Respiratorio de Oriente Medio/aislamiento & purificación , Filogenia , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación
15.
J Virol ; 95(18): e0085321, 2021 08 25.
Artículo en Inglés | MEDLINE | ID: mdl-34232065

RESUMEN

Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus causing acute intestinal infection in pigs, with high mortality often seen in neonatal pigs. The newborns rely on innate immune responses against invading pathogens because of lacking adaptive immunity. However, how PEDV disables the innate immunity of newborns toward severe infection remains unknown. We found that PEDV infection led to reduced expression of histone deacetylases (HDACs), especially HDAC1, in porcine IPEC-J2 cells. HDACs are considered important regulators of innate immunity. We hypothesized that PEDV interacts with certain host factors to regulate HDAC1 expression in favor of its replication. We show that HDAC1 acted as a negative regulator of PEDV replication in IPEC-J2 cells, as shown by chemical inhibition, gene knockout, and overexpression. A GC-box (GCCCCACCCCC) within the HDAC1 promoter region was identified for Sp1 binding in IPEC-J2 cells. Treatment of the cells with Sp1 inhibitor mithramycin A inhibited HDAC1 expression, indicating direct regulation of HDAC1 expression by Sp1. Of the viral proteins that were overexpressed in IPEC-J2 cells, the N protein was found to be present in the nuclei and more inhibitory to HDAC1 transcription. The putative nuclear localization sequence 261PKKNKSR267 contributed to its nuclear localization. The N protein interacted with Sp1 and interfered with its binding to the promoter region, thereby inhibiting its transcriptional activity for HDAC1 expression. Our findings reveal a novel mechanism of PEDV evasion of the host responses, offering implications for studying the infection processes of other coronaviruses. IMPORTANCE The enteric coronavirus porcine epidemic diarrhea virus (PEDV) causes fatal acute intestinal infection in neonatal pigs that rely on innate immune responses. Histone deacetylases (HDACs) play important roles in innate immune regulation. Our study found PEDV suppresses HDAC1 expression via the interaction of its N protein and porcine Sp1, which identified a novel mechanism of PEDV evasion of the host responses to benefit its replication. This study suggests that other coronaviruses, including SARS-CoV and SARS-CoV-2, also make use of their N proteins to intercept the host immune responses in favor of their infection.


Asunto(s)
Infecciones por Coronavirus/veterinaria , Células Epiteliales/virología , Histona Desacetilasa 1/antagonistas & inhibidores , Mucosa Intestinal/virología , Factor de Transcripción Sp1/metabolismo , Enfermedades de los Porcinos/virología , Proteínas no Estructurales Virales/metabolismo , Replicación Viral , Animales , Células Cultivadas , Infecciones por Coronavirus/metabolismo , Infecciones por Coronavirus/virología , Células Epiteliales/metabolismo , Células Epiteliales/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Virus de la Diarrea Epidémica Porcina/patogenicidad , Factor de Transcripción Sp1/genética , Porcinos , Enfermedades de los Porcinos/metabolismo , Enfermedades de los Porcinos/patología , Proteínas no Estructurales Virales/genética
16.
Vet Microbiol ; 260: 109151, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34237662

RESUMEN

Avian infectious bronchitis virus (IBV), belonging to Gammacoronavirus, is an economically important respiratory virus affecting poultry industry worldwide. The virus can infect chickens at all ages, whereas young chickens (less than 15 day old) are more susceptible to it. The present study was conducted to investigate effects of dietary supplementation of black soldier fly (Hermetia illucens L.) larvae (BSFL) on immune responses in IBV infected 10-day-old chickens. BSFL were ground to powder and mixed with commercial fodder (1%, 5%, and 10 % [mass] BSFL powder) to feed 1-day-old yellow broilers for ten days and then challenged with IBV. Our results indicated that commercial fodder supplemented with 10 % BSFL [mass] reduced mortalities (20 %) and morbidities (80 %), as well as IBV viral loads in tracheas (65.8 %) and kidneys (20.4 %) from 3-day post challenge (dpc), comparing to that of IBV-infected chickens fed with non-additive commercial fodder. Furthermore, at 3-day post challenge (dpc), 10 % BSFL [mass] supplemented chickens presented more CD8+ T lymphocytes in peripheral blood and a rise in interferon-g (IFN-γ) at both mRNA and protein levels in spleens, comparing with chickens fed with commercial fodder. Furthermore, the mRNA abundance of MHC-I, Fas, LITAF, and IL-2 in the spleens of 10 % BSFL [mass] supplemented chickens increased at different time points after challenge. The present results suggest that supplemental BSFL could improve CD8+ T lymphocytes proliferation, thus benefit young chickens to defend against IBV infection.


Asunto(s)
Linfocitos T CD8-positivos/inmunología , Pollos/fisiología , Infecciones por Coronavirus/veterinaria , Dípteros/fisiología , Virus de la Bronquitis Infecciosa/inmunología , Enfermedades de las Aves de Corral/dietoterapia , Alimentación Animal/análisis , Animales , Linfocitos T CD8-positivos/citología , Pollos/inmunología , Pollos/virología , Infecciones por Coronavirus/dietoterapia , Infecciones por Coronavirus/inmunología , Dieta/veterinaria , Virus de la Bronquitis Infecciosa/genética , Larva , Masculino , Enfermedades de las Aves de Corral/inmunología
17.
Vet Microbiol ; 260: 109179, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34271305

RESUMEN

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of coronavirus disease 2019 (COVID-19), has rapidly spread worldwide. Studies of transmission of the virus carried out in animals have suggested that certain animals may be susceptible to infection with SARS-CoV-2. The aim of the present study was to investigate the infection of SARS-CoV-2 in pets (18 cats and 20 dogs) from owners previously confirmed as COVID-19-positive. Oropharyngeal and rectal swabs were taken and analyzed by real-time RT-PCR assays, while blood samples were taken for antibody detection. Of the total pets analyzed, one cat was found reactive to SARS-CoV-2 by real-time RT-PCR of an oropharyngeal and a rectal swab. This cat presented only sneezing as a clinical sign. Serological analysis confirmed the presence of antibodies in the serum sample from this cat, as well as in the serum from another cat non-reactive to real-time RT-PCR. Complete sequence and phylogenetic analysis allowed determining that the SARS-CoV-2 genome belonged to the B.1.499 lineage. This lineage has been reported in different provinces of Argentina, mainly in the Metropolitan Area of Buenos Aires. This study notifies the first detection of the natural infection and molecular analysis of SARS-CoV-2 in a cat from Argentina whose owner where COVID-19-positive. Although there is currently no evidence that cats can spread COVID-19, results suggest that health authorities should test pets with COVID-19-positive owners.


Asunto(s)
Enfermedades de los Gatos/virología , Infecciones por Coronavirus/veterinaria , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , Animales , Argentina , Prueba de Ácido Nucleico para COVID-19/veterinaria , Enfermedades de los Gatos/diagnóstico , Gatos , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/virología , ADN Complementario/química , Perros , Femenino , Genoma Viral/genética , Secuenciación de Nucleótidos de Alto Rendimiento/veterinaria , Filogenia , ARN Viral/genética , ARN Viral/aislamiento & purificación , SARS-CoV-2/clasificación
18.
Arch Virol ; 166(9): 2461-2468, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34212242

RESUMEN

Bovine coronavirus (BCoV) can be spread by animal activity. Although cattle farming is widespread in Turkey, there are few studies of BCoV. The aim of this study was to evaluate the current situation regarding BCoV in Turkey. This is the first study reporting the full-length nucleotide sequences of BCoV spike (S) genes in Turkey. Samples were collected from 119 cattle with clinical signs of respiratory (n = 78) or digestive tract (n = 41) infection on different farms located across widely separated provinces in Turkey. The samples were screened for BCoV using RT-nested PCR targeting the N gene, which identified BCoV in 35 samples (9 faeces and 26 nasal discharge). RT-PCR analysis of the S gene produced partial/full-length S gene sequences from 11 samples (8 faeces and 3 nasal discharge samples). A phylogenetic tree of the S gene sequences was made to analyze the genetic relationships among BCoVs from Turkey and other countries. The results showed that the local strains present in faeces and nasal discharge samples had many different amino acid changes. Some of these changes were shown in previous studies to be critical for tropism. This study provides new data on BCoV in Turkey that will be valuable in designing effective vaccine approaches and control strategies.


Asunto(s)
Enfermedades de los Bovinos/epidemiología , Infecciones por Coronavirus/veterinaria , Coronavirus Bovino/genética , Diarrea/veterinaria , ARN Viral/genética , Infecciones del Sistema Respiratorio/veterinaria , Glicoproteína de la Espiga del Coronavirus/genética , Agricultura , Sustitución de Aminoácidos , Animales , Bovinos , Enfermedades de los Bovinos/virología , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/virología , Coronavirus Bovino/clasificación , Diarrea/epidemiología , Diarrea/virología , Monitoreo Epidemiológico/veterinaria , Evolución Molecular , Heces/virología , Humanos , Mutación , Cavidad Nasal/virología , Filogenia , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Turquia/epidemiología
19.
Virus Res ; 302: 198497, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34217778

RESUMEN

Porcine deltacoronavirus (PDCoV) is an emerging coronavirus that causes vomiting, diarrhea, dehydration, and even death of piglets, resulting in significant losses to the pig industry worldwide. However, the epitopes of PDCoV remain largely unknown. In this study, a monoclonal antibody (mAb) against the PDCoV nucleocapsid (N) protein, termed 9G1, was prepared using the lymphocyte hybridoma technique, and was identified as a type IgG1 with a κ light chain and reacted with the native N protein of PDCoV. Furthermore, the epitope recognized by the 9G1 mAb was subjected to western blot and an ELISA using truncated recombinant proteins and synthetic polypeptides of the PDCoV N protein. The results indicate that 9G1 mAb recognized the epitope, G59TPIPPSYAFYY70 (EP-9G1), a novel linear B cell epitope of the PDCoV N protein. A comparison analysis revealed that the EP-9G1 epitope was highly conserved among PDCoV strains, in which four residues (G59-F68YY70) were observed among different coronavirus genera. These data demonstrate that the EP-9G1 epitope identified in this study provides some basic information for further characterization of the antigenic structure of the PDCoV N protein and has potential use for developing diagnostic reagents for PDCoV.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Infecciones por Coronavirus/veterinaria , Deltacoronavirus/inmunología , Epítopos de Linfocito B/inmunología , Proteínas de la Nucleocápside/inmunología , Secuencia de Aminoácidos , Animales , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/virología , Deltacoronavirus/genética , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Inmunoglobulina G/inmunología , Ratones Endogámicos BALB C , Proteínas de la Nucleocápside/genética , Proteínas Recombinantes , Alineación de Secuencia , Porcinos
20.
Vet Microbiol ; 259: 109155, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34197977

RESUMEN

Turkey coronavirus (TCoV) can cause a highly contagious enteric disease in turkeys with severe economic losses in the global turkey industry. To date, no commercial vaccines are available for control of the disease. In the present study, we isolated a field strain (NC1743) of TCoV and evaluated its pathogenicity in specific-pathogen-free (SPF) turkey poults to establish a TCoV disease model. The results showed that the TCoV NC1743 isolate was pathogenic to turkey poults with a minimal infectious dose at 106 EID50/bird. About 50 % of one-day-old SPF turkeys infected with the virus's minimal infectious dose exhibited typical enteric disease signs and lesions from 6 days post-infection (dpi) to the end of the experiment (21 dpi). In contrast, fewer than 20 % of older turkeys (1- or 2-week-old) infected with the same amount of TCoV displayed enteric disease signs, which disappeared after 15-18 dpi. Although all infected turkeys, regardless of age, shed TCoV, the older turkeys shed less virus than the younger birds, and 50 % of the 2-week-old birds even cleared the virus at 21 dpi. Furthermore, the viral infection caused day-old turkeys more body-weight-gain reduction than older birds. The overall data demonstrated that the TCoV NC1743 isolate is a highly pathogenic strain and younger turkeys are more susceptible to TCoV infection than older birds. Thus, one-day-old turkeys infected with the minimal infectious dose of TCoV NC1743 could be used as a TCoV disease model to study the disease pathogenesis, and the TCoV NC1743 strain could be used as a challenge virus to evaluate a vaccine protective efficacy.


Asunto(s)
Infecciones por Coronavirus/veterinaria , Coronavirus del Pavo/patogenicidad , Enfermedades de las Aves de Corral/prevención & control , Pavos/virología , Animales , Anticuerpos Antivirales/sangre , Infecciones por Coronavirus/sangre , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/virología , Coronavirus del Pavo/clasificación , Modelos Animales de Enfermedad , Enfermedades de las Aves de Corral/sangre , Enfermedades de las Aves de Corral/virología , Organismos Libres de Patógenos Específicos
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