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1.
Res Vet Sci ; 109: 121-128, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27892860

ABSTRACT

This study describes the changes observed in the placentas of mice experimentally infected with an abortigenic strain of EHV-1 at mid-pregnancy and euthanized at days 3 and 4 post-infection. We analyzed microscopic vascular alterations, cell proliferation and death by immunohistochemistry, and the expression of IFN-γ, TNF-α and the IL-10 by qPCR and flow cytometry. Infected mice showed slight respiratory signs and ruffled fur during the first two days post-infection. Virus isolation and DNA detection were positive only in the lungs of the infected mice. Vascular congestion, increase in the labyrinth area, and a significant reduction in fetal capillary endothelium surface of infected placentas were found. Cell proliferation was significantly reduced in the infected placentas, whereas the apoptosis was significantly increased. IL10, TNF and IFN-γ showed different expression in the infected placentas and uteri. The effects of EHV-1 during pregnancy depend on different pathogenic mechanisms in which vascular alterations, and cell death and proliferation and local cytokine changes are compromised.


Subject(s)
Abortion, Veterinary/pathology , Cell Death , Cell Proliferation , Cytokines/genetics , Herpesviridae Infections/veterinary , Abortion, Veterinary/virology , Animals , Cytokines/metabolism , Female , Flow Cytometry , Herpesviridae Infections/pathology , Herpesviridae Infections/virology , Herpesvirus 1, Equid/physiology , Interferon-gamma/genetics , Interferon-gamma/metabolism , Interleukin-10/genetics , Interleukin-10/metabolism , Mice , Mice, Inbred BALB C , Placenta/pathology , Placenta/virology , Pregnancy , Real-Time Polymerase Chain Reaction , Specific Pathogen-Free Organisms , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Uterus/pathology , Uterus/virology
2.
J Comp Pathol ; 151(4): 384-93, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25124330

ABSTRACT

Equine herpesvirus (EHV)-1 induces respiratory infection, neurological disorders and abortion in horses. Most of the currently available attenuated or inactivated vaccines against this infection are administered intramuscularly and only provide partial protection against the respiratory disease. The present study examines the effect of intranasal immunization with purified EHV-1 recombinant glycoprotein D (gD) in BALB/c mice followed by challenge with three different EHV-1 strains during early to mid-pregnancy. The induced viral infection was evaluated by virus isolation, DNA detection by polymerase chain reaction, histopathology and immunohistochemical localization of antigen in the lung, placenta and uterus. Non-immunized mice showed clinical signs of infection, positive virus isolation from lungs and uteri, and abortion induced by one of the virus strains. Endometrial lesions developed in some of these animals that have been described previously only in horses. Immunized mice and their offspring had no viral infection or typical lesions. Intranasally administered gD therefore induced partial or complete protection against three different EHV-1 strains in BALB/c mice.


Subject(s)
Herpesviridae Infections/veterinary , Herpesvirus 1, Equid , Vaccination/methods , Viral Envelope Proteins/administration & dosage , Administration, Intranasal , Animals , Disease Models, Animal , Female , Herpesviridae Infections/prevention & control , Immunohistochemistry , Mice , Mice, Inbred BALB C , Polymerase Chain Reaction , Pregnancy , Recombinant Proteins/administration & dosage , Recombinant Proteins/immunology , Viral Envelope Proteins/immunology
3.
Res Vet Sci ; 95(2): 709-16, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23602434

ABSTRACT

Bovine herpesvirus (BoHV) type 1.1 (BoHV-1.1) causes repeated outbreaks of upper respiratory disease and abortion in cattle. The systemic effects of BoHV-1.1 in rabbits, using intranasal inoculation are reported. Female rabbits were divided into four groups and inoculated with the virus 10 days before mating, and at 15 or 22 days of pregnancy. Studies of the clinical signs, antibody production, virus isolation, and DNA detection as well as histological and immunohistochemical studies were carried out on lungs, kidneys, spleen, placentas, uteri and foetal tissues. All virus-inoculated animals developed respiratory clinical signs and a humoral response. BoHV-1.1 was isolated from nasal swabs and plasma rich in leukocytes, and viral DNA was detected in blood, dead foetuses and placentas. Histopathological lesions were found in the respiratory tract and some placentas and foetuses were immunohistochemically positive. Intranasal inoculation might be useful to study the systemic effects of BoHV-1.1 infection in the rabbit model.


Subject(s)
Herpesviridae Infections/veterinary , Herpesvirus 1, Bovine , Rabbits , Animals , Antibodies, Viral/blood , Female , Herpesviridae Infections/virology , Lung/pathology , Polymerase Chain Reaction , Pregnancy , Turbinates/pathology
4.
Vet Pathol ; 50(5): 849-56, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23381927

ABSTRACT

Mice are commonly used as an experimental model to investigate the Equid herpesvirus 1 (EHV-1) infection. This model easily reproduces the disease, and the clinical signs are more or less similar to those observed in the horse, the natural host. During natural infection, the acute course of respiratory infection is mandatory for the development of adaptive immune response. Since interactions between EHV-1 and anesthetics are possible, the study investigated whether the early events of murine pulmonary immune response could be affected by different anesthetics. Therefore, mice were experimentally infected with a unique EHV-1 strain under the effects of ether, ketamine/xylazine, or isoflurane. Clinical signs and histopathological lesions in the lungs were described, and the cell death and proliferation rates of sham-inoculated or infected animals were quantified using immunohistochemistry. Clinical signs were more severe in animals anesthetized with ether. Qualitative differences in the recruited inflammatory cells were observed following application of anesthesia. The level of infection between the infected groups was not statistically significant. However, lungs from ketamine/xylazine-anesthetized animals showed the highest cell death rates, whereas those from isoflurane-anesthetized animals showed the highest proliferation rates. It has been emphasized that anesthetics alone or their interactions with EHV-1 modify the response against the infection. An appropriate selection of the anesthetic during experimental studies is relevant to minimize wrong conclusions.


Subject(s)
Anesthetics/pharmacology , Disease Models, Animal , Herpesviridae Infections/immunology , Herpesviridae Infections/pathology , Herpesvirus 1, Equid , Lung/pathology , Analysis of Variance , Animals , Cell Death/drug effects , Cell Proliferation/drug effects , Immunohistochemistry , Isoflurane , Ketamine , Lung/drug effects , Mice , Xylazine
5.
Rev Argent Microbiol ; 41(4): 207-11, 2009.
Article in English | MEDLINE | ID: mdl-20085182

ABSTRACT

This work reports a method for rapid amplification of the complete genome of equine influenza virus subtype 2 (H3N8). A ThermoScript reverse transcriptase instead of the avian myeloblastosis virus reverse transcriptase or Moloney murine leukemia virus reverse transcriptase was used. This enzyme has demonstrated higher thermal stability and is described as suitable to make long cDNA with a complex secondary structure. The product obtained by this method can be cloned, used in later sequencing reactions or nested-PCR with the purpose of achieving a rapid diagnosis and characterization of the equine influenza virus type A. This detection assay might be a valuable tool for diagnosis and screening of field samples as well as for conducting molecular studies.


Subject(s)
Genome, Viral , Influenza A Virus, H3N8 Subtype/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Animals , Consensus Sequence , Conserved Sequence , Influenza A Virus, H3N8 Subtype/classification , Orthomyxoviridae Infections/diagnosis , Orthomyxoviridae Infections/veterinary , Orthomyxoviridae Infections/virology , RNA, Viral/genetics , RNA-Directed DNA Polymerase , Sequence Alignment , Sequence Homology, Nucleic Acid
6.
Virus Genes ; 38(1): 113-7, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19023651

ABSTRACT

Equid Herpesvirus 1 (EHV-1) has long been causally implicated in the occurrence of abortion, neonatal death, respiratory disease, and neurological disorders in horses. This study analyzed for the first time the characteristics of the genomic section of Argentinian EHV-1 strains and reconstructed the phylogeny in order to establish their origin. The phylogenetic dataset included 22 Argentinian strains and four additional reference strains isolated in other countries. The intergenic region between ORF 62 and ORF 63 was amplified by PCR and sequenced. The phylogenetic analysis carried out by parsimony algorithms showed that six of the Argentinian strains had the same origin as British and Japanese strains. The mapping of symptoms caused by EHV-1 suggested that neonatal disease developed through convergent evolution, which would constitute an adaptation mechanism of the virus. This study constitutes the first analysis carried out in South-American strains that establishes the phylogenetic relationship between Argentinian strains and rebuilds the evolutionary history of symptoms. This study focuses on a very important aspect of evolution of Herpesviridae infecting perissodactyls and attempts to shed light on the evolution of symptoms, an issue of high clinical interest.


Subject(s)
Herpesvirus 1, Equid/classification , Herpesvirus 1, Equid/genetics , Phylogeny , Animals , Argentina , Cluster Analysis , DNA, Viral/chemistry , DNA, Viral/genetics , Herpesvirus 1, Equid/isolation & purification , Horses , Molecular Epidemiology , Molecular Sequence Data , Polymerase Chain Reaction/methods , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid
7.
Rev Sci Tech ; 28(3): 1085-90, 2009 Dec.
Article in English | MEDLINE | ID: mdl-20462167

ABSTRACT

Equine herpesvirus 1 (EHV-1) is a major cause of epidemic abortion, neonatal mortality, respiratory disease and neurological disorders in horses. In South America, the virus has been isolated in Brazil, Argentina and Colombia. In Chile pathological findings from one aborted foetus have been reported, and in Uruguay only serological data about EHV-1 activity have been found. Some pathological findings were reported in Uruguay several years ago, but these data have never been officially confirmed. The present work describes the relevant findings of a study of EHV-1 infections in the Uruguayan equine population using polymerase chain reaction (PCR) and histological and immunohistochemical analysis techniques. The sequence analysis of a portion of the glycoprotein C gene amplified by PCR confirmed EHV-1 activity. The real-time PCR revealed the association of the virus with the non-neuropathogenic genotype. This study describes for the first time the immunohistochemical and molecular detection of EHV-1 in Uruguay.


Subject(s)
Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/isolation & purification , Horse Diseases/diagnosis , Immunohistochemistry/veterinary , Polymerase Chain Reaction/veterinary , Aborted Fetus/pathology , Aborted Fetus/virology , Abortion, Veterinary/virology , Animals , Herpesviridae Infections/diagnosis , Herpesviridae Infections/microbiology , Herpesviridae Infections/pathology , Herpesvirus 1, Equid/genetics , Horse Diseases/microbiology , Horse Diseases/pathology , Horses , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/microbiology , Respiratory Tract Infections/pathology , Respiratory Tract Infections/veterinary , Uruguay
8.
Article in English | MEDLINE | ID: mdl-17062117

ABSTRACT

Equine herpesvirus 1 (EHV-1) was first isolated in Argentina in 1979. This strain SPv has special restriction patterns, but a previous study demonstrated that SPv did not modify its growth in cell culture. In addition, it showed low virulence in the mouse respiratory model consistently with results found in female BALB/C at different state of gestation. This study evaluates in a mouse respiratory model, if primary infection with SPv strain protects animals from subsequent challenge with a pathogenic strain. Body weight loss was not observed in mice intranasally inoculated with SPv strain and challenged with HH1 Japanese strain. The SPv primary infection does not completely prevent clinical presentation by HH1 infection but the SPv inoculated animals recovered more quickly, with less intense and less persistent histological lesions. The challenge infection caused a rapid and prolonged increase in anti-EHV-1 antibodies in the mice previously infected with SPv, along with a more rapid reduction of viral titres in lungs. In this work it was demonstrated that this EHV-1 strain constitute a good immunogen. These results show that this SPv strain could be considered to produce an EHV-1 vaccine.


Subject(s)
Disease Models, Animal , Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/genetics , Herpesvirus 1, Equid/pathogenicity , Horse Diseases/virology , Polymorphism, Restriction Fragment Length , Animals , Antibodies, Viral/blood , Argentina , Herpesviridae Infections/virology , Horses , Mice , Mice, Inbred BALB C , Random Allocation , Species Specificity , Specific Pathogen-Free Organisms , Virulence/genetics
9.
Rev Sci Tech ; 25(3): 1075-9, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17361771

ABSTRACT

To determine the genomic variation of equine herpesviruses (EHVs) isolated in Argentina between 1979 and the first half of 2004, DNA sequences from all 69 strains isolated were analysed. Sixty strains were recovered from aborted fetuses, one from leucocyte-rich plasma from a horse with respiratory signs and eight from cases of neonatal disease. The DNA was extracted from rabbit kidney epithelial (RK13) cells infected with each strain and digested with three restriction endonucleases (BamHI, Bg/II and KpnI). Two strains could be differentiated using BamHI restriction and were assigned to the EHV-1 1B prototype group. Only one of these two strains was typed EHV-1 1B with Bg/II. DNA digestion with KpnI was ineffective. The results obtained in this study demonstrate that the EHV-1 1B genome has been present in Argentina since at least 1996. The finding of two strains with this electropherotype suggests that there is genomic heterogeneity among Argentinian isolates.


Subject(s)
Abortion, Veterinary/virology , DNA, Viral/analysis , Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/genetics , Horse Diseases/virology , Pregnancy Complications, Infectious/veterinary , Animals , Argentina/epidemiology , Base Sequence , DNA Restriction Enzymes , Female , Genetic Variation , Genome , Herpesviridae Infections/diagnosis , Herpesviridae Infections/epidemiology , Herpesviridae Infections/virology , Herpesvirus 1, Equid/classification , Herpesvirus 1, Equid/isolation & purification , Horse Diseases/diagnosis , Horse Diseases/epidemiology , Horses , Pregnancy , Pregnancy Complications, Infectious/diagnosis , Pregnancy Complications, Infectious/epidemiology , Pregnancy Complications, Infectious/virology , Restriction Mapping/methods , Restriction Mapping/veterinary
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