RESUMEN
In-vitro fertilization is a routine livestock-breeding technique widely used around the world. Several studies have reported the interaction of bovine viral-diarrhea virus (BVDV) with gametes and in-vitro-produced (IVP) bovine embryos. Since, gene expression in BVDV-infected IVP bovine embryos is scarcely addressed. The aim of this work was to evaluate the differential expression of genes involved in immune and inflammatory response. Groups of 20-25 embryos on Day 6 (morula stage) were exposed (infected) or not (control) to an NCP-BVDV strain in SOF medium. After 24 h, embryos that reached expanded blastocyst stage were washed. Total RNA of each embryo group was extracted to determine the transcription levels of 9 specific transcripts related with antiviral and inflammatory response by SYBR Green real time quantitative (RT-qPCR). Culture media and an aliquot of the last embryos wash on Day 7 were analyzed by titration and virus isolation, respectively. A conventional PCR confirmed BVDV presence in IVP embryos. A significantly higher expression of interferon-α was observed in blastocysts exposed to NCP-BVDV compared to the controls (p < 0.05). In this study, the upregulation of INFα and TLR7 genes involved in inflammatory and immune response in BVDV-infected IVP bovine embryos is a new finding in this field. This differential expression suggest that embryonic cells could function in a manner like immune cells by recognizing and responding early to interaction with viral pathogens. These results provide new insights into the action of BVDV on the complex molecular pathways controlling bovine early embryonic development.
Asunto(s)
Diarrea Mucosa Bovina Viral , Bovinos , Virus de la Diarrea Viral Bovina/inmunología , Desarrollo Embrionario/inmunología , Expresión Génica/inmunología , Interferón-alfa , Animales , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/inmunología , Diarrea Mucosa Bovina Viral/virología , Bovinos/embriología , Bovinos/inmunología , Virus de la Diarrea Viral Bovina/aislamiento & purificación , Embrión de Mamíferos/inmunología , Embrión de Mamíferos/virología , Femenino , Fertilización In Vitro , Interferón-alfa/inmunología , Receptor Toll-Like 7/inmunologíaRESUMEN
The aim of the report was to present the circulation of BVDV (bovine viral diarrhea virus) in the cattle population and determine the cause of the failure of vaccination failure leading to the birth of the PI (persistently infected) calf. The case study was carried out at the BVDV-free animal breeding center and cattle farm, where the vaccination program against BVDV was implemented in 2012, and each newly introduced animal was serologically and virologically tested for BVDV. In this case, a blood sample was taken from a 9-month-old breeding bull. Positive RT-PCR and negative ELISA serology results were obtained. The tests were repeated at 2-week intervals, and the results confirmed the presence of the virus and the absence of specific antibodies, i.e., persistent infection. Additionally, sequencing and phylogenetic analysis were performed, and the BVDV-1d subgenotype was detected. The results of this study showed that pregnant heifers and cows that are vaccinated multiple times with the killed vaccine containing BVDV-1a may not be fully protected against infection with other subgenotypes of BVDV, including their fetuses, which can become PI calves.
Asunto(s)
Diarrea Mucosa Bovina Viral/prevención & control , Virus de la Diarrea Viral Bovina/inmunología , Enfermedades Fetales/prevención & control , Vacunas Virales/administración & dosificación , Vacunas Virales/inmunología , Animales , Anticuerpos Antivirales/sangre , Diarrea Mucosa Bovina Viral/sangre , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/virología , Bovinos , Virus de la Diarrea Viral Bovina/clasificación , Virus de la Diarrea Viral Bovina/genética , Virus de la Diarrea Viral Bovina/aislamiento & purificación , Femenino , Enfermedades Fetales/virología , Masculino , Infección Persistente/sangre , Infección Persistente/virología , Filogenia , Embarazo , Vacunación , Vacunas de Productos Inactivados/administración & dosificación , Vacunas de Productos Inactivados/genética , Vacunas de Productos Inactivados/inmunología , Vacunas Virales/genéticaRESUMEN
Bovine viral diarrhea virus (BVDV) infection occurs in the cattle population worldwide. Non-cytopathic (ncp) BVDV strains cause transient infection (TI) or persistent infection (PI) depending on the host's immune status. Immunocompetent adult animals and fetuses in late gestation resolve the infection. Fetal infection in early gestation results in PI with chronic viremia and life-long viral shedding, ensuring virus perpetuation in the population. Eighteen pregnant heifers, divided into three groups, were intranasally inoculated with ncp BVDV2 virus early (day 75) and late (day 175) in gestation, or kept BVDV-naïve. Fetuses were retrieved on day 190. Antiviral activity in blood of dams and fetuses, maternal expression of interferon (IFN) stimulated gene 15kDa (ISG15), virological and serological status of heifers and fetuses, and fetal growth were studied. A pronounced antiviral activity in blood of heifers and TI fetuses during acute BVDV infection was accompanied by drastic up-regulation of ISG15 mRNA in maternal blood. Only one PI fetus expressed low IFN response 115 days post inoculation despite high BVDV antigen and RNA levels. PI fetuses presented with growth retardation. Infection of pregnant heifers with ncp BVDV2 early in gestation adversely affects fetal development and antiviral responses, despite protective immune responses in the dam.
Asunto(s)
Diarrea Mucosa Bovina Viral/inmunología , Virus de la Diarrea Viral Bovina Tipo 2/inmunología , Enfermedades Fetales/veterinaria , Factores Reguladores del Interferón/genética , Interferón Tipo I/inmunología , Preñez/inmunología , Aborto Veterinario/virología , Animales , Anticuerpos Antivirales/sangre , Antígenos Virales/sangre , Bioensayo , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/virología , Bovinos , Efecto Citopatogénico Viral , Virus de la Diarrea Viral Bovina Tipo 2/patogenicidad , Femenino , Desarrollo Fetal , Enfermedades Fetales/inmunología , Feto , Factores Reguladores del Interferón/sangre , Interferón Tipo I/sangre , Embarazo , ARN Viral/sangre , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
In the first experiment, heifers were infected experimentally with bovine viral diarrhea virus type II (BVDV-type II, strain CD87; characterized by high morbidity and mortality). Subsequently, in vitro fertilized embryos were produced from oocytes collected on Day 4, 8, and 16 post infection. In a total of 29 heifers, the infectious virus was detected in 55% of the samples of the follicular fluid, in 10% of the oviductal cells, in 10% of the uterine flushes and in 41% of the in vitro fertilized embryos. The highest number of embryos associated with the virus was detected in the group of animals slaughtered on Day 8 post infection (58%). The amount of the virus (10(1.5-2.0) TCID50/mL) associated with the washed single embryos generated from oocytes of heifers 8 and 16 d post infection was sufficient for disease transmission by intravenous inoculation to the seronegative recipients (6/15). In the second experiment, uninfected oocytes were exposed in vitro to BVDV (10(5) TCID50/mL) in the maturation medium and then fertilized and cultured prior to viral assay. Virus was detected in 4 of 7 samples containing embryos but not in samples of embryos produced from the control group of uninfected oocytes. The presence of BVDV in the IVF system did not affect embryonic development in vitro. In conclusion, it appears that BVDV-type II has the ability to be transferred with oocytes through the IVF system, resulting in infectious embryos with normal morphological appearance which may have a potential for disease transmission.
Asunto(s)
Diarrea Mucosa Bovina Viral/embriología , Bovinos/embriología , Virus de la Diarrea Viral Bovina/patogenicidad , Fertilización In Vitro/veterinaria , Animales , Diarrea Mucosa Bovina Viral/patología , Diarrea Mucosa Bovina Viral/transmisión , Bovinos/fisiología , Técnicas de Cocultivo , Virus de la Diarrea Viral Bovina/aislamiento & purificación , Embrión de Mamíferos/virología , Trompas Uterinas/virología , Femenino , Líquido Folicular/virología , Masculino , Pruebas de Neutralización/veterinaria , Oocitos/virología , Embarazo , Útero/virologíaRESUMEN
BVDV shares with other Pestiviruses the ability to cross the placenta of pregnant host animals. The effects of fetal infections are complex and depend on a number of factors, e.g., age of the zygote/embryo stage, no infection seems to occur. During the last one third of gestation the infection is terminated by the ontogeny of the fetal immune system. This leaves a window of susceptibility during early stages of fetal development allowing establishment of viral persistence and/or the development of a number of fetopathologic effects. Additionally, fertility problems and abortions are observed. Calves that are born immunotolerant to BVDV and persistently viremic display a wide variety of abnormalities. However, there is an unknown proportion of calves born without any clinical signs indicative of persistent infection. The time of fetal infection during the first stages of pregnancy seems to play a crucial role with respect to the lesions induced. Generally, early infections seem to induce less damage compared with late infections, suggesting an indirect, possibly immune-mediated pathogenesis. Additionally, direct virus-cell interactions may play a role. Few data exist about the influence of differences in viral virulence on fetal pathology. Likewise the role of the viral target cell range is not clear.
Asunto(s)
Diarrea Mucosa Bovina Viral/complicaciones , Virus de la Diarrea Viral Bovina/fisiología , Enfermedades Fetales/veterinaria , Complicaciones Infecciosas del Embarazo/veterinaria , Animales , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/fisiopatología , Bovinos , Enfermedades del Sistema Nervioso Central/embriología , Enfermedades del Sistema Nervioso Central/veterinaria , Enfermedades del Sistema Nervioso Central/virología , Femenino , Enfermedades Fetales/fisiopatología , Enfermedades Fetales/virología , Infertilidad/fisiopatología , Infertilidad/veterinaria , Infertilidad/virología , Embarazo , Complicaciones Infecciosas del Embarazo/fisiopatología , Complicaciones Infecciosas del Embarazo/virologíaRESUMEN
Losses resulting from pestivirus infections in cattle are of considerable importance to the livestock industry yet, until relatively recently, they have been poorly understood. The escalation of research effort in this field has started to clarify the pathogenesis and epidemiology of the associated diseases but the evolution of understanding is far from complete. Sufficient information exists to indicate the means by which pestivirus infection is introduced to and maintained in populations of cattle. A brief review of current knowledge is given placing particular emphasis on the role played by persistent infections which have resulted from intrauterine infection. The crux of the problem of control is seen to be the avoidance of fetal infection in early gestation. Ways of achieving this which include the immunisation of female cattle with live or inactivated virus vaccines are discussed and areas requiring further work are indicated.
Asunto(s)
Diarrea Mucosa Bovina Viral/transmisión , Enfermedades de los Bovinos/transmisión , Animales , Anticuerpos Antivirales/análisis , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/prevención & control , Bovinos , Transferencia de Embrión , Femenino , Enfermedades Fetales/veterinaria , Masculino , Embarazo , Semen/microbiología , VacunasRESUMEN
The events which followed the introduction of a heifer into a dairy herd were consistent with the animal being persistently infected with bovine diarrhoea-mucosal disease virus. Obvious damage was limited to the progeny of cows which were in the first 168 days of gestation at that time. Only fetuses up to 81 days of gestation at the putative time of introduction of infection became persistently infected in calfhood and, although they exhibited body tremor, two such calves necropsied at three months of age lacked macroscopic or microscopic lesions in the central nervous system. In contrast calves which had been more advanced in gestation, at 146 and 153 days at the time of infection, had eliminated the infection and had lesions of cerebellar dysplasia and multifocal retinal atrophy.
Asunto(s)
Diarrea Mucosa Bovina Viral/embriología , Enfermedades de los Bovinos/embriología , Aborto Séptico/etiología , Aborto Veterinario/etiología , Animales , Diarrea Mucosa Bovina Viral/complicaciones , Diarrea Mucosa Bovina Viral/transmisión , Bovinos , Enfermedades de los Bovinos/etiología , Cerebelo/patología , Anomalías Congénitas/patología , Anomalías Congénitas/veterinaria , Femenino , Edad Gestacional , Embarazo , Retina/patologíaRESUMEN
Paraffin sections from various organs of sheep fetuses following transplacental infection with non-cytopathogenic (ncp) bovine viral diarrhoea virus (BVDV) or cytopathogenic (cp) BVDV were stained immunohistochemically with BVDV-specific monoclonal antibodies. Comparison of the distribution of viral antigen in sections from fetuses of experiment A revealed that in organs such as parotid, thyroid, thymus, lung, spleen, kidney, liver and skin from 20 days post inoculation (p.i.) onwards numerous antigen-containing cells were present. In organs of fetuses infected with cp BVDV, however, antigen-positive cells were only detectable until days 10 and 14 p.i. These findings suggest that the ncp BVDV used in experiment A replicated considerably faster and more efficient than the cp BVDV used in experiment B and that the two virus biotypes differ considerably concerning their tropism for fetal ovine organs.
Asunto(s)
Diarrea Mucosa Bovina Viral/embriología , Feto/virología , Pestivirus/clasificación , Animales , Antígenos Virales/análisis , Bovinos , Femenino , Inmunohistoquímica/métodos , Especificidad de Órganos , Pestivirus/aislamiento & purificación , Pestivirus/fisiología , Embarazo , Ovinos , Replicación ViralRESUMEN
The objective was to determine whether a multivalent modified-live virus vaccine containing noncytopathic bovine viral diarrhea virus (BVDV) administered off-label to pregnant cattle can result in persistently infected fetuses and to assess whether vaccinal strains can be shed to unvaccinated pregnant cattle commingling with vaccinates. Nineteen BVDV-naïve pregnant heifers were randomly assigned to two groups: cattle vaccinated near Day 77 of gestation with modified-live virus vaccine containing BVDV-1a (WRL strain), bovine herpes virus-1, parainfluenza 3, and bovine respiratory syncytial virus (Vx group; N = 10) or control unvaccinated cattle (N = 9). During the course of the study a voluntary stop-sale/recall was conducted by the manufacturer because of the presence of a BVDV contaminant in the vaccine. At Day 175 of gestation, fetuses were removed by Cesarean section and fetal tissues were submitted for virus isolation, and quantitative reverse transcription polymerase chain reaction using BVDV-1- and BVDV-2-specific probes. Nucleotide sequencing of viral RNA was performed for quantitative reverse transcription polymerase chain reaction-positive samples. Two vaccinated and two control heifers aborted their pregnancies, but their fetuses were unavailable for BVDV testing. Virus was isolated from all eight fetuses in the Vx group heifers and from 2 of 7 fetuses in the control unvaccinated heifers. Only BVDV-2 was detected in fetuses from the Vx group, and only BVDV-1 was detected in the two fetuses from the control group. Both BVDV-1 and BVDV-2 were detected in the vaccine. In conclusion, vaccination of pregnant heifers with a contaminated modified-live BVDV vaccine resulted in development of BVDV-2 persistently infected fetuses in all tested vaccinated animals. Furthermore, BVDV was apparently shed to unvaccinated heifers causing fetal infections from which only BVDV-1 was detected.
Asunto(s)
Diarrea Mucosa Bovina Viral/etiología , Virus de la Diarrea Viral Bovina Tipo 1/inmunología , Contaminación de Medicamentos , Enfermedades Fetales/etiología , Transmisión Vertical de Enfermedad Infecciosa , Complicaciones Infecciosas del Embarazo/etiología , Vacunación/efectos adversos , Vacunas Atenuadas/efectos adversos , Animales , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/inmunología , Diarrea Mucosa Bovina Viral/prevención & control , Bovinos , Virus de la Diarrea Viral Bovina Tipo 1/aislamiento & purificación , Femenino , Enfermedades Fetales/sangre , Enfermedades Fetales/inmunología , Transmisión Vertical de Enfermedad Infecciosa/prevención & control , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Uso Fuera de lo Indicado , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/prevención & control , Vacunación/veterinaria , Vacunas Atenuadas/inmunología , Viremia/transmisión , Viremia/veterinariaRESUMEN
The objective of this study was to chronologically investigate the effect of bovine viral diarrhea virus (BVDV) infection on early pregnant cows before placenta formation. Six cows were intravenously inoculated with either BVDV (treated, n=4) or growth medium (control, n=2) (day 0) at day 26 of pregnancy. Two treated cows and one control cow were euthanized on day 3 post-infection and the remaining animals were euthanized on day 6. BVDV was isolated from maternal tissues such as lymphoid or reproductive tissues of treated animals on days 3 and 6 post-infection. Additionally, one treated cow autopsied on day 6 post-infection had evidence of infectious BVDV in the allantoic membranes, allantoic fluid and embryos. In three treated cows, a significant decline in progesterone concentration was also observed post-infection while in control cows they remained constant. Therefore, BVDV can infect bovine embryos before placenta formation and may affect progesterone profiles in cows during early pregnancy.
Asunto(s)
Diarrea Mucosa Bovina Viral/virología , Virus de la Diarrea Viral Bovina/fisiología , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Animales , Diarrea Mucosa Bovina Viral/embriología , Diarrea Mucosa Bovina Viral/orina , Bovinos , Membranas Extraembrionarias/virología , Femenino , Tejido Linfoide/virología , Ovario/virología , EmbarazoAsunto(s)
Diarrea Mucosa Bovina Viral/embriología , Bovinos/anomalías , Enfermedades Fetales/patología , Animales , Animales Recién Nacidos , Diarrea Mucosa Bovina Viral/patología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Resultado Fatal , Enfermedades Fetales/embriología , Masculino , Prevalencia , Estudios SeroepidemiológicosRESUMEN
Bovine viral diarrhea virus (BVDV) can associate with in vitro fertilized (IVF) bovine embryos despite washing and trypsin treatment. An antiviral compound, DB606 (2-(4-[2-imidazolinyl]phenyl)-5-(4-methoxyphenyl)furan), inhibits the replication of BVDV in bovine uterine tubal epithelial cells, Madin Darby bovine kidney cells, and fetal fibroblast cells. As well, DB606 in in vitro culture medium does not affect embryonic development. Antiviral-treated-IVF embryos placed into recipients developed into clinically normal calves. The objective of this project was to determine if these resultant heifer calves were capable of reproducing. Seven heifers from each of the treatment groups (natural breeding, IVF embryo, and IVF embryo cultured in DB606) of the previous study were used. At 20-27 months of age, the heifers were exposed to a fertile bull in a single pasture during a 63 d breeding season. Five of the seven heifers originating from natural breeding were pregnant 35 d after removal of the bull and calved. All of the heifers resulting from transfer of untreated IVF embryos were pregnant at 35 d; however, one aborted the fetus at 5-7 months of gestation. All of the heifers derived from transfer of IVF embryos cultured in DB606 were pregnant and calved. Offspring from dams of all treatment groups were clinically normal at birth. Adjusted 205 d weaning weights were not significantly different among the offspring of the treated and untreated dams. These results indicate that culture of bovine-IVF embryos in DB606 does not impair future reproductive capacity of resulting heifers.
Asunto(s)
Antivirales/farmacología , Diarrea Mucosa Bovina Viral/embriología , Bovinos , Embrión de Mamíferos/efectos de los fármacos , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Reproducción/efectos de los fármacos , Animales , Antivirales/uso terapéutico , Diarrea Mucosa Bovina Viral/tratamiento farmacológico , Bovinos/embriología , Bovinos/fisiología , Virus de la Diarrea Viral Bovina , Eficiencia/efectos de los fármacos , Técnicas de Cultivo de Embriones/veterinaria , Embrión de Mamíferos/fisiología , Embrión de Mamíferos/virología , Femenino , Fertilización In Vitro/métodos , Furanos/farmacología , Furanos/uso terapéutico , Imidazolinas/farmacología , Imidazolinas/uso terapéutico , Embarazo , Índice de Embarazo , Efectos Tardíos de la Exposición Prenatal/veterinaria , Efectos Tardíos de la Exposición Prenatal/virología , Reproducción/fisiologíaRESUMEN
Eighteen pregnant Merino ewes were inoculated intravenously between days 65 and 68 of gestation with the unpurified cytopathogenic (cp) bovine virus diarrhoea virus (BVDV) strain Indiana (experiment I). In experiment II, three ewes were inoculated with the same virus after two successive plaque isolations in order to compare its pathogenicity for the fetus with special regard to lesions in the fetal brain. In experiment I, fetal blood and tissue samples, allantoic fluids and placentomes were collected sequentially between 10 and 80 days post-inoculation (p.i.). BVDV was recovered from 6 of 19 fetuses examined during the first 3 weeks after inoculation. From fetuses sampled between 30 and 50 days p.i. virus was isolated from three cases only, and from 60 days p.i. onwards virus was no longer recovered. BVDV was longer detected in the allantoic fluid than in fetal tissues and continued to be present until 80 days post-inoculation. From tissue samples of two fetuses of experiment I, only non-cytopathogenic BVDV was isolated, whilst samples from seven fetuses contained the cp BVDV biotype as revealed by an immunoplaque assay. The cp biotype was also isolated from placentomes. In experiment II, virus was not isolated from any of the tissue samples of two living fetuses collected at 67 days post-inoculation. In both experiments, cp BVDV was recovered from allantoic fluid samples. In contrast to the developing fetal brain, other tissues or organs seemed to be less vulnerable to the cp BVDV strain Indiana. The partial purification of this virus strain did not affect its pathogenicity for the brains of the developing fetuses.
Asunto(s)
Diarrea Mucosa Bovina Viral/embriología , Virus de la Diarrea Viral Bovina/aislamiento & purificación , Feto/virología , Complicaciones Infecciosas del Embarazo , Enfermedades de las Ovejas/embriología , Alantoides/virología , Animales , Anticuerpos Antivirales/sangre , Diarrea Mucosa Bovina Viral/virología , Bovinos , Virus de la Diarrea Viral Bovina/inmunología , Femenino , Feto/patología , Embarazo , Complicaciones Infecciosas del Embarazo/virología , Ovinos , Enfermedades de las Ovejas/virologíaRESUMEN
Despite the importance of congenital viral infections in both veterinary and human medicine, only limited experimental work has been carried out to elucidate the mechanisms involved in transplacental virus infections. To further an understanding of fetal infection with pestiviruses, the distribution of bovine pestivirus in the uterine and fetal tissues of ewes in early pregnancy, following a natural route of infection, was investigated. On the 18th day of pregnancy, nine ewes were inoculated by the intranasal route with 1 x 10(5) 50% tissue culture infective doses of an Australian isolate of noncytopathic bovine pestivirus (bovine viral diarrhea virus genotype 1). All ewes were ovariohysterectomized at approximately 100 hours postinfection. Samples from the reproductive tract and conceptus were examined histologically and tested for bovine pestivirus by nested reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry and for interferon-tau mRNA expression by nonnested RT-PCR. Although no histopathologic changes were observed in the maternal or fetal tissues, virus was detected in the reproductive tract of all nine ewes and in all of the conceptuses examined. At the time of surgery, only two of the nine ewes were demonstrably viremic. This study demonstrates that bovine pestivirus can spread from a natural site of infection to the ovine fetus within 4 days in the absence of maternal immunity and despite the presence of interferon expression in the reproductive tract.