RESUMEN
Severe fever with thrombocytopenia syndrome (SFTS) is caused by infection with Dabie bandavirus [formerly SFTS virus (SFTSV)] and is an emerging zoonotic disease. Dogs can be infected with SFTSV, but its pathogenicity and transmissibility have not been fully elucidated. In experiment 1, immunocompetent dogs were intramuscularly inoculated with SFTSV. In experiment 2, immunosuppressed dogs (immunosuppressed group; oral azathioprine 5 mg/kg/day for 30 days) were intramuscularly inoculated with SFTSV. Both immunosuppressed and immunocompetent contact dogs were co-housed with the SFTSV-inoculated dogs that had been immunosuppressed. Immunocompetent SFTSV-infected dogs did not show any clinical symptom. However, immunosuppressed SFTSV-infected dogs showed high fever and weight loss without lethality. In all SFTSV-infected dogs, viral RNA could be measured in the serum only after 3 days post infection (DPI) and neutralizing antibodies were detected in the serum beginning 9 DPI. SFTSV shedding in the urine and faeces of some infected dogs occurred between 4 and 6 DPI. The immunocompromised SFTSV-infected dogs showed thrombocytopenia beginning 3 DPI to the end of the experiment (24 DPI). We confirmed SFTSV transmission to one of three immunocompetent co-housed dogs. This dog showed a high fever, weight loss, and shed viral RNA by urine. Viral RNA and neutralizing antibodies were also detected in the serum. These results demonstrated that intramuscular inoculation with SFTSV induced minor clinical symptoms in dogs, and intraspecies SFTSV transmission in dogs can occur by contact.
Asunto(s)
Infecciones por Bunyaviridae , Enfermedades de los Perros , Síndrome de Trombocitopenia Febril Grave , Animales , Anticuerpos Neutralizantes , Azatioprina , Infecciones por Bunyaviridae/veterinaria , Perros , Phlebovirus , ARN Viral , Síndrome de Trombocitopenia Febril Grave/veterinaria , Virulencia , Pérdida de PesoRESUMEN
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonotic disease, which causes high fever, thrombocytopenia, and death in humans and animals in East Asian countries. The pathogenicity of SFTS virus (SFTSV) remains unclear. We intraperitoneally infected three groups of mice: wild-type (WT), mice treated with blocking anti-type I interferon (IFN)-α receptor antibody (IFNAR Ab), and IFNAR knockout (IFNAR-/-) mice, with four doses of SFTSV (KH1, 5 × 105 to 5 × 102 FAID50). The WT mice survived all SFTSV infective doses. The IFNAR Ab mice died within 7 days post-infection (dpi) with all doses of SFTSV except that the mice were infected with 5 × 102 FAID50 SFTSV. The IFNAR-/- mice died after infection with all doses of SFTSV within four dpi. No SFTSV infection caused hyperthermia in any mice, whereas all the dead mice showed hypothermia and weight loss. In the WT mice, SFTSV RNA was detected in the eyes, oral swabs, urine, and feces at 5 dpi. Similar patterns were observed in the IFNAR Ab and IFNAR-/- mice after 3 dpi, but not in feces. The IFNAR Ab mice showed viral shedding until 7 dpi. The SFTSV RNA loads were higher in organs of the IFNAR-/- mice compared to the other groups. Histopathologically, coagulation necrosis and mononuclear inflammatory cell infiltration in the liver and white pulp atrophy in the spleen were seen as the main lesions in the IFN signaling lacking mice. Immunohistochemically, SFTSV antigens were mainly detected in the marginal zone of the white pulp of the spleen in all groups of mice, but more viral antigens were observed in the spleen of the IFNAR-/- mice. Collectively, the IFN signaling-deficient mice were highly susceptible to SFTSV and more viral burden could be demonstrated in various excreta and organs of the mice when IFN signaling was inhibited.
RESUMEN
To date, the implications of interleukin 6 (IL-6) for immune responses in the context of Brucella infection are still unknown. In the present study, we found that Brucella abortus infection induced marked production of IL-6 in mice that was important for sufficient differentiation of CD8+ T cells, a key factor in Brucella clearance. Blocking IL-6 signaling also significantly induced serum IL-4 and IL-10, together with a decreased gamma interferon (IFN-γ) level, suggesting that IL-6 is essential for priming the T-helper (Th) 1 cell immune response during Brucella infection. The IL-6 pathway also activated the bactericidal activity of primary and cultured macrophages. Bacterial killing was markedly abrogated when IL-6 signaling was suppressed, and this phenomenon was mainly associated with decreased activity of lysosome-mediated killing. Interestingly, suppressor of cytokine signaling 3 (SOCS3) was important for regulating the IL-6-dependent anti-Brucella activity through the JAK/STAT pathway. During early infection, in the absence of SOCS3, IL-6 exhibited anti-inflammatory effects and lysosome-mediated killing inhibition; however, the increase in SOCS3 successfully shifted functional IL-6 toward proinflammatory brucellacidal activity in the late stage. Our data clearly indicate that IL-6 contributes to host resistance against B. abortus infection by controlling brucellacidal activity in macrophages and priming cellular immune responses.
Asunto(s)
Brucella abortus/fisiología , Citocinas/metabolismo , Interleucina-6/metabolismo , Macrófagos/microbiología , Animales , Anticuerpos , Células Presentadoras de Antígenos , Receptor gp130 de Citocinas/genética , Receptor gp130 de Citocinas/metabolismo , Citocinas/genética , Interleucina-6/genética , Ratones , Células RAW 264.7 , Interferencia de ARN , Receptores de Interleucina-6/genética , Receptores de Interleucina-6/metabolismo , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT1/metabolismo , Factor de Transcripción STAT3/genética , Factor de Transcripción STAT3/metabolismo , Transducción de Señal , Proteína 3 Supresora de la Señalización de Citocinas/genética , Proteína 3 Supresora de la Señalización de Citocinas/metabolismo , Células TH1/metabolismoRESUMEN
In this study, we aimed to assess trends in antimicrobial resistance and to investigate the characteristics of extended-spectrum ß-lactamase (ESBL)-producing isolates from bovine mastitic milk from 2012 to 2015. A total of 374 Escherichia coli isolates were analyzed (154 in 2012, 113 in 2013, 76 in 2014, and 31 in 2015). No consistent trends in antimicrobial resistance of E. coli isolates occurred during the 4-yr period. The most frequently observed resistance was tetracycline (23.3%), followed by streptomycin (17.1%), ampicillin (16.6%), neomycin (11.8%), and trimethoprim/sulfamethoxazole (11.2%). Multidrug resistance was observed in 15.5% of isolates. Among these isolates, 15 (4.0%) carried one or more blaCTX-M and AmpC ESBL genes from 11 different farms, including blaCTX-M-15 at 4 farms, blaCTX-M-3 at 2 farms, blaCTX-M-1 at 3 farms, and blaCMY-2 at 3 farms. This study is the first report of blaCTX-M-3-producing E. coli in dairy milk. Transfer of ESBL was observed in 3 blaCTX-M-3-producing isolates, 1 blaCTX-M-1-producing isolate, and all 3 blaCMY-2-producing isolates. Almost all blaCTX-M-15 and blaCTX-M-1 genes possessed an insertion sequence, ISECP1, upstream of the blaCTX-M gene. Identical pulsed-field gel electrophoresis profiles were also observed in blaCTX-M-producing E. coli from the same farm. These results suggested that ESBL might spread by both clonal and horizontal spread in dairy farms in South Korea. Although no significant changes occurred in the antimicrobial resistance of E. coli during the 4-yr study period, the resistance rates and presence of ESBL were high compared with those in other countries. Thus, these findings suggest the importance of control measures for E. coli, particularly ESBL-producing bacteria, on dairy farms to reduce treatment failure and transmission to humans.
Asunto(s)
Escherichia coli/aislamiento & purificación , beta-Lactamasas , Animales , Antiinfecciosos/uso terapéutico , Bovinos , Infecciones por Escherichia coli/veterinaria , Humanos , Leche/microbiología , PlásmidosRESUMEN
Five outbreaks of foot-and-mouth disease have occurred in South Korea during 2000-2011. Macro-analysis of these outbreaks showed a correlation with outbreaks in countries in eastern Asia. Genetic analyses of food-and-mouth disease viruses in South Korea showed a correlation with viruses that are prevalent in neighboring countries.
Asunto(s)
Enfermedades Transmisibles Emergentes , Virus de la Fiebre Aftosa , Fiebre Aftosa/epidemiología , Animales , Asia Sudoriental/epidemiología , Brotes de Enfermedades , Asia Oriental/epidemiología , Fiebre Aftosa/historia , Virus de la Fiebre Aftosa/clasificación , Virus de la Fiebre Aftosa/genética , Geografía Médica , Historia del Siglo XXI , Humanos , Ganado , República de Corea/epidemiología , Factores de RiesgoRESUMEN
Foot-and-mouth disease (FMD) is a highly contagious infectious disease, and the use of vaccines is known to be effective for its prevention. In 2010/2011, there was an epidemic of the South East Asia (SEA) topotype in East Asian countries. We adapted the SEA topotype virus isolated in November 2010 in Korea in cells to analyze the characteristics of the virus and evaluate its possibility as a vaccine. After cell culture adaptation, the FMD virus particle 146S was purified to develop an inactivated oil vaccine for SEA or other topotypes. To measure its immunogenicity, pigs were inoculated with the experimental vaccine at different concentrations of the antigen. The results indicated that the groups immunized with at least 7.5 µg antigen were protected from homologous challenge. The immunized pigs were also protected against heterologous virus (ME-SA topotype) challenge. The genetic variations between the two field isolates and the adapted vaccine strains were identified in six amino acids by complete genome sequencing.
Asunto(s)
Fiebre Aftosa/prevención & control , Enfermedades de los Porcinos/prevención & control , Vacunas Virales/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Asia Sudoriental/epidemiología , Fiebre Aftosa/epidemiología , Virus de la Fiebre Aftosa/clasificación , Virus de la Fiebre Aftosa/genética , Genoma Viral , Sus scrofa/inmunología , Porcinos , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/virología , Vacunas de Productos Inactivados/inmunologíaRESUMEN
Chronic wasting disease (CWD) is a naturally occurring prion disease in North American deer (Odocoileus species), Rocky mountain elk (Cervus elaphus nelsoni) and moose (Alces alces). The disease was first confirmed in the Republic of Korea in 2001, and subsequent cases were diagnosed in 2004, 2005 and 2010. The experimental host range of CWD includes ferrets, several species of voles, white-footed mice, deer mice and Syrian golden hamsters. In addition, CWD was transmitted to the transgenic mouse over-expressing elk or deer prion protein efficiently, but not to wild type mouse. Here, we report the experimental transmission of elk CWD to conventional VM/Dk mice reaching 100% attack rate after second passage. The CWD-prion-affected wild type mice will be a useful model for future CWD studies.
Asunto(s)
Encéfalo/patología , Modelos Animales de Enfermedad , Ratones Endogámicos , Enfermedad Debilitante Crónica/fisiopatología , Animales , Inmunohistoquímica , Ratones , República de Corea , Especificidad de la Especie , Enfermedad Debilitante Crónica/transmisiónRESUMEN
Chronic wasting disease (CWD) has been recognized as a naturally occurring prion disease in North American deer (Odocoileus species), Rocky Mountain elk (Cervus elaphus nelsoni) and moose (Alces alces). The disease was confirmed only in elk in the Republic of Korea in 2001, 2004 and 2005. Epidemiological investigations showed that CWD was introduced via importation of infected elk from Canada between 1994 and 1997. In spite of the increasing geographic distribution and host range of CWD, little is known about the prion strain (s) responsible for distinct outbreaks of the disease. We carried out strain characterization, using transgenic mice overexpressing elk prion protein, including clinical assessment, pathological examination and biochemical analyses, in brain tissues derived following primary through tertiary transmissions. The final incubation period was shortened to approximately 130 dpi due to adaptation. Biochemical profiles remained identical between passages. Lesion profiling in recipient mice brains showed similar patterns of vacuolation scores and intensity. It is clear that there were no biochemical or histopathological differences in Korean CWD cases in 2001 and 2004, suggesting a single strain was responsible for the outbreaks.
Asunto(s)
Encéfalo/patología , Ciervos , Brotes de Enfermedades/veterinaria , Priones/genética , Enfermedad Debilitante Crónica/epidemiología , Enfermedad Debilitante Crónica/metabolismo , Animales , Western Blotting/veterinaria , Encéfalo/metabolismo , Ratones , Ratones Transgénicos , Priones/metabolismo , República de Corea/epidemiología , Enfermedad Debilitante Crónica/patologíaRESUMEN
Elk prion protein (PrP(C)) has been confirmed to be capable of rendering rabbit epithelial RK13 cells permissive to temporal infection by chronic wasting disease (CWD) prions. The present study satisfactorily generated persistently CWD prion-affected RK13 cells (RKC1-11) using elk PrP(C) expressing cells (elkRK13) that were generated via the lentiviral expression system with high efficiency. The elkRK13 cells have been shown to be permissive to accumulation of abnormal isoforms of prion protein (PrP(Sc)) resulting from CWD prions up to 97 serial passages thus far. This novel prion-affected cell line will help facilitate investigation of the molecular basis of CWD prion pathogenesis and confirmation of CWD prion infectivity in vitro.
Asunto(s)
Línea Celular , Ciervos/metabolismo , Lentivirus/metabolismo , Priones/metabolismo , Enfermedad Debilitante Crónica/metabolismo , Animales , Vectores Genéticos/metabolismo , Lentivirus/genética , Priones/genética , Conejos , República de CoreaRESUMEN
Aino, Akabane and Chuzan viruses are arthropod-borne (arbo) viruses transmitted by blood-sucking insects like mosquitoes and Culicoides biting midges. These arbovirus infections are mainly associated with abortion, stillbirth and congenital defects in pregnant cattle, sheep and goats, which induces a considerable economic loss in livestock industry. The viruses seem to be widely distributed in Southeast Asia and Australia. As a control strategy, an inactivated trivalent vaccine against Aino, Akabane and Chuzan virus was developed by using binary ethylenimine or formalin as an inactivating agent. The newly developed trivalent vaccine is evaluated for its safety and immunogenicity in animals such as mice, guinea pigs and cattle. The immune responses were significantly detected within 2-weeks after second vaccination without any side effects. Since the field application of experimental vaccine also revealed increased antibodies in inoculated cattle, we demonstrated that these trivalent vaccines could be used as a vaccine to control the arboviral infections in ruminants.
Asunto(s)
Orthobunyavirus/inmunología , Virus Palyam/inmunología , Vacunas Virales/aislamiento & purificación , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Infecciones por Bunyaviridae/inmunología , Infecciones por Bunyaviridae/prevención & control , Infecciones por Bunyaviridae/veterinaria , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/prevención & control , Femenino , Formaldehído , Enfermedades de las Cabras/inmunología , Enfermedades de las Cabras/prevención & control , Cabras , Cobayas , Ratones , Ratones Endogámicos ICR , Orthobunyavirus/patogenicidad , Virus Palyam/patogenicidad , Embarazo , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/prevención & control , Infecciones por Reoviridae/veterinaria , Ovinos , Enfermedades de las Ovejas/inmunología , Enfermedades de las Ovejas/prevención & control , Vacunas de Productos Inactivados/administración & dosificación , Vacunas de Productos Inactivados/aislamiento & purificación , Vacunas Virales/administración & dosificaciónRESUMEN
Bovine leukemia virus (BLV) envelope glycoprotein (gp51/ gp30(T-)), consisting of BLV gp51 and BLV gp30 that lacked its C-terminal transmembrane domain, was expressed in insect cells under the control of the baculovirus polyhedron promoter. Recombinant BLV gp51/gp30(T-) secreted from insect cells was determined by immunofluorescence, enzyme-linked immunosorbent and western blot assays using a BLV-specific monoclonal antibody and BLV-positive bovine antibodies. An agar gel immunodiffusion (AGID) test using gp51/gp30(T-) as the antigen for the detection of BLV antibodies in serum was developed and compared to traditional AGID, which uses wild type BLV antigen derived from fetal lamb kidney cells. AGID with the recombinant BLV gp51/gp30(T-) was relatively more sensitive than traditional AGID. When the two methods were tested with bovine sera from the field, the recombinant BLV gp51/gp30(T-) and traditional antigen had a relative sensitivity of 69.8% and 67.4%, respectively, and a relative specificity of 93.3% and 92.3%. These results indicated that the recombinant BLV gp51/gp30(T-) is an effective alternative antigen for the diagnosis of BLV infection in cattle.
Asunto(s)
Baculoviridae/metabolismo , Regulación Viral de la Expresión Génica/fisiología , Inmunodifusión/veterinaria , Virus de la Leucemia Bovina/metabolismo , Proteínas del Envoltorio Viral/metabolismo , Agar , Animales , Anticuerpos Antivirales/sangre , Antígenos Virales/inmunología , Bovinos , Línea Celular , Leucosis Bovina Enzoótica/sangre , Leucosis Bovina Enzoótica/inmunología , Inmunodifusión/métodos , Riñón/citología , Virus de la Leucemia Bovina/genética , Biología Molecular , Ovinos , Proteínas del Envoltorio Viral/genéticaRESUMEN
Transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) are major etiological agents of diarrhea and death in piglets. Multiplex real-time reverse transcriptase (RT)-PCR was developed for simultaneous differential quantification of each virus in a single reaction tube, using Cy5- and FAM-labeled TaqMan-probes based on sequences from the TGEV and PEDV nucleocapsid genes. The copy numbers for transcripts of TGEV and PEDV were quantified using this assay over a range from 9x10(7) to 9x10(1) copies and 7x10(7) to 7x10(1) copies, respectively. The variability of the intra-assay and inter-assay were evaluated using standard solutions of each transcript, with coefficients of variation (CV) less than 3.43 and 3.33%, respectively. Piglets were experimentally infected with virulent TGEV and PEDV, and the amounts of virus from the onset of diarrhea were measured. Samples obtained from farms experiencing PED or TGE were quantified between 10(2) and 10(5) RNA copies. In conclusion, this assay provides an effective etiological diagnostic tool for detecting and quantifying viral loads. The assay may also prove useful for detecting infections, ultimately leading to better disease control on farms.
Asunto(s)
Diarrea/veterinaria , Virus de la Diarrea Epidémica Porcina/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Enfermedades de los Porcinos/virología , Virus de la Gastroenteritis Transmisible/aislamiento & purificación , Carga Viral/métodos , Animales , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Diarrea/virología , Heces/virología , Gastroenteritis Porcina Transmisible/virología , Sensibilidad y Especificidad , PorcinosRESUMEN
Vector-borne arboviruses produce mild to severe symptoms in domestic animals. Bovine ephemeral fever (BEF), Akabane, Aino, and Chuzan virus have been primarily attributed to reproductive disorders or febrile diseases in cattle, and Japanese encephalitis virus (JEV) is mainly associated with reproductive failures in swine. We investigated antibody titers from domestic swine against four bovine arboviruses (BEF, Akabane, Aino, and Chuzan virus) and from cattle against JEV in Korea. While the positive rates for Akabane and BEF were 37.4% and 15.7%, the positive incidence of Chuzan and Aino were relatively low, with positive rates of 3.04% and 0.4%, respectively, based on a virus neutralization assay. Antibody titers against more than one virus were also frequently detected in domestic swine. The incidence of JEV was 51.3% among domestic cattle. In addition, one positive case was detected in the thoracic fluids from 35 aborted calves, based on the hemagglutination inhibition test. Our results indicate that swine are susceptible hosts of bovine arboviruses without showing clinical symptoms in a natural environment. Moreover, we confirmed that JEV could be associated with reproductive failure in pregnant cattle, as were other vector-borne bovine arboviruses assessed in this study.
Asunto(s)
Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/virología , Virus de la Encefalitis Japonesa (Especie)/inmunología , Encefalitis Japonesa/veterinaria , Virus de la Fiebre Efímera Bovina/inmunología , Fiebre Efímera/epidemiología , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/virología , Animales , Anticuerpos Antivirales/sangre , Bovinos , Encefalitis Japonesa/sangre , Encefalitis Japonesa/epidemiología , Encefalitis Japonesa/virología , Fiebre Efímera/sangre , Fiebre Efímera/virología , Pruebas de Hemaglutinación , Incidencia , Corea (Geográfico)/epidemiología , Pruebas de Neutralización , PorcinosRESUMEN
Akabane, Aino and Chuzan virus are arthropod-borne (arbo) viruses mainly associated with reproductive failures in cattle. We investigated apoptosis in Vero cells (C-1586) infected with Akabane, Aino and Chuzan virus. The fragmentation of chromosomal DNA was simultaneously detected with the progress of cytopathic effect from 48 hr to 72 hr post infection, depending on viruses. Although the treatment of cycloheximide blocked apoptosis in Vero cells infected with three viruses, actinomycin D did not prevent DNA oligomerization, thus indicating that de novo viral protein synthesis is critical for viral apoptosis. In addition, the activation of caspase-3 was also detected in Vero cells by indirect fluorescent assay. From the present results, it is of future interest whether apoptotic characteristics of these viruses are related to pathogenecity in vivo.