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1.
Arch Virol ; 168(4): 122, 2023 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-36977931

RESUMO

The ORF 70 gene of equid alphaherpesvirus type 3 (EHV-3) encodes glycoprotein G (gG), which is conserved in the majority of alphaherpesviruses. This glycoprotein is located in the viral envelope and has the characteristic of being secreted into the culture medium after proteolytic processing. It modulates the antiviral immune response of the host by interacting with chemokines. The aim of this study was to identify and characterize EHV-3 gG. By constructing viruses with HA-tagged gG, it was possible to detect gG in lysates of infected cells, their supernatants, and purified virions. A 100-, 60-, and 17-kDa form of the protein were detected in viral particles, while a 60-kDa form was identified in supernatants of infected cells. The role of EHV-3 gG in the viral infection cycle was assessed by the construction of a gG-minus EHV-3 mutant and its gG-positive revertant. When growth characteristics in an equine dermal fibroblast cell line were compared, the plaque size and the growth kinetics of the gG-minus mutant were similar to those of the revertant virus, suggesting that EHV-3 gG does not play a role in direct cell-to-cell transmission or virus proliferation of EHV-3 in tissue culture. The identification and characterization of EHV-3 gG described here provide a solid background for further studies to assess whether this glycoprotein has a function in modulating the host immune response.


Assuntos
Infecções por Herpesviridae , Herpesvirus Equídeo 1 , Herpesvirus Equídeo 3 , Animais , Cavalos , Proteínas do Envelope Viral/metabolismo , Herpesvirus Equídeo 1/genética , Herpesvirus Equídeo 3/metabolismo , Linhagem Celular , Glicoproteínas/genética
2.
Vet Med Sci ; 9(2): 717-720, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36253808

RESUMO

Equine coital rash (ECE) is a highly contagious benign infection that induces lesions on external genitals, and it is caused by the equine herpesvirus type 3 (EHV-3). Although the disease is globally distributed, its presence in Chile has not been documented from a genetic point of view. Here, we performed polymerase chain reaction screenings for EHV-3 in lesions of external genitals in four horses belonging to a riding station at Bulnes, Ñuble Region, Chile. We sequenced a fragment of the glycoprotein G (gG) gene from three horses with clinical signs of ECE. The sequences were identical between them and 99.7% similar to a haplotype of EHV-3 detected in Brazil, and phylogenetically related with homologue from Japan, Russia and Brazil. Our results show the presence of EHV-3 for the first time in horses with ECE in Chile.


Assuntos
Infecções por Herpesviridae , Herpesvirus Equídeo 3 , Doenças dos Cavalos , Animais , Cavalos , Chile/epidemiologia , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/veterinária , Reação em Cadeia da Polimerase/veterinária , Sequência de Bases , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/epidemiologia
3.
Microbiol Spectr ; 9(2): e0098321, 2021 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-34668724

RESUMO

Distinct but related species of elephant endotheliotropic herpesviruses (EEHVs) circulate within Asian and African elephant populations. Primary infection with EEHVs endemic among Asian elephants can cause clinical illness and lethal EEHV hemorrhagic disease (EEHV-HD). The degree to which this occurs among African elephants has not been fully established. Recent cases of EEHV-HD caused by the EEHV3 species in African elephants housed in North American zoos has heightened concern about the susceptibility of this elephant species to EEHV-HD. In this study, we utilize the luciferase immunoprecipitation system (LIPS) to generate a serological assay specific for EEHV3 in African elephants by detecting antibodies against the EEHV3 E34 protein. The results showed that the majority of tested elephants from four separate and genetically unrelated herds, including five elephants that survived clinical illness associated with EEHV3, were positive for prior infection with EEHV3. However, African elephants who succumbed to EEHV3-HD were seronegative for EEHV3 prior to lethal infection. This supports the hypothesis that fatal EEHV-HD caused by EEHV3 is associated with primary infection rather than reactivation of latent virus. Lastly, we observed that African elephants, like Asian elephants, acquire abundant anti-EEHV antibodies prenatally and that anti-EEHV3 specific antibodies were either never detected or declined to undetectable levels in those animals that died from lethal disease following EEHV3 infection. IMPORTANCE Prior to 2019, only five cases of clinical disease from EEHV infection among African elephants had been documented. Since 2019, there have been at least seven EEHV-HD cases in North American zoos, resulting in three fatalities, all associated with EEHV3. Evidence is accumulating to suggest that EEHV-associated clinical illness and death among Asian elephants is due to primary infection and may be associated with waning anti-EEHV antibody levels in young elephants. The development of the EEHV3 serological test described in this study enabled us to confirm that similar dynamics may be contributing to EEHV-HD in African elephants. The ability to screen for EEHV immune status in African elephant calves will have a major impact on managing captive African elephant herds and will provide new tools for investigating and understanding EEHV in wild populations.


Assuntos
Elefantes/virologia , Transtornos Hemorrágicos/veterinária , Herpesvirus Equídeo 3/imunologia , Zoonoses Virais/diagnóstico , Zoonoses Virais/mortalidade , Animais , Animais de Zoológico/virologia , Anticorpos Antivirais/sangue , Feminino , Transtornos Hemorrágicos/diagnóstico , Transtornos Hemorrágicos/virologia , Herpesvirus Equídeo 3/patogenicidade , Masculino , Testes Sorológicos , Zoonoses Virais/patologia
4.
Acta Vet Scand ; 63(1): 6, 2021 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-33531030

RESUMO

Equine coital exanthema (ECE) caused by equid alphaherpesvirus 3 (EHV-3) is a contagious venereal disease. It is characterized by the formation of papules, vesicles, pustules and ulcers on the external genitals of both mares and stallions. The Icelandic horse is the only breed in Iceland and has lived isolated in the country for over 1000 years. Three types of equine herpesviruses (EHV) have been found in Iceland, EHV-4, EHV-2 and EHV-5, while EHV-1 has never been detected. Symptoms resembling ECE have previous been observed in horses in Iceland, arousing suspicion of EHV-3 infection, but this has never been confirmed using virological methods. Samples were collected from a mare with papules on the vulva and inoculated in primary equine kidney cells. Cytopathic effects developed as rounded cells and syncytial formation. Polymerase chain reaction and sequencing of the partial glycoprotein G and DNA polymerase genes identified the isolated virus as EHV-3. On the basis of the findings, EHV-3 infection was verified for the first time in the native Icelandic horse population.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/isolamento & purificação , Doenças dos Cavalos/diagnóstico , Animais , Diagnóstico Diferencial , Feminino , Infecções por Herpesviridae/diagnóstico , Doenças dos Cavalos/virologia , Cavalos , Islândia
5.
J Equine Vet Sci ; 94: 103199, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33077066

RESUMO

Equid alphaherpesvirus 3 (EHV-3) is the etiological agent of equine coital exanthema (ECE). Because no vaccines or antiviral therapies are available, prevention consists of clinical examination of mares and stallions before mating or semen collection and resting from breeding activities when lesions are present. However, this methodology does not identify subclinically infected animals. Ganciclovir is the most potent compound known to reduce EHV-3 replication. This study aimed to evaluate the efficacy of topical ganciclovir application to reduce EHV-3 replication in experimentally infected mares. A pilot study, after a double-blind completely randomized design, was carried out. Twenty mares were randomly divided into five groups (three treated with ganciclovir with different regimen of doses, one treated with a placebo, and one nontreated). Mares were experimentally infected with EHV-3 on day 0. Rectal temperature, clinical signs, and lesions were recorded. Daily perineal and vaginal swabs were evaluated by quantitative polymerase chain reaction for virus detection. The antibody response was assessed by a virus neutralization test in serum samples collected weekly. Mares experimentally infected with EHV-3 and treated with ganciclovir twice a day for 13 days showed reduced levels and duration of viral excretion and less severe lesions. The viral excretion period was reduced from 18 to nine days compared with the untreated groups. We concluded that ganciclovir had an antiviral effect on EHV-3 replication when topically administered in mares showing clinical signs of ECE. Further trials should be performed to optimize the dose of the antiviral for a definitive formulation.


Assuntos
Exantema , Herpesvirus Equídeo 3 , Doenças dos Cavalos , Animais , Exantema/veterinária , Feminino , Ganciclovir , Cavalos , Masculino , Projetos Piloto
6.
J Vet Med Sci ; 82(9): 1299-1305, 2020 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-32684614

RESUMO

In 2017, two Thoroughbred stallions, A and B in Farms A and B, respectively, in Hokkaido in Japan showed clinical signs of equine coital exanthema (ECE). In 2020, stallion C in Farm B showed clinical signs of ECE. Eighteen mares were mated within five days before stallion A developed ECE. Ten mares that mated within 3 days before onset showed clinical signs of ECE on the external genitalia. Equine herpesvirus 3 (EHV-3) was isolated from vaginal swabs from three mares that mated within 2 days before onset. Swabs from 12 mares that mated within 4 days before onset were real-time PCR (rPCR)-positive and nine of those mares had an increased EHV-3 antibody titer. The three stallions were administered valaciclovir orally and topical acyclovir ointment was applied. Treatment started on the next day after onset in stallion A and on the day of onset in stallions B and C. EHV-3 was firstly isolated from penis swabs of stallions A and B before treatment and from penis swabs of stallion C 2 days after treatment. EHV-3 was not isolated after 8, 5 and 8 days from onset in stallions A, B and C, respectively. However, swabs were rPCR-positive for at least 12, 9 and 15 days after onset of stallions A, B and C, respectively. EHV-3 was excreted from the stallions at least within 4 days before the onset of ECE, and acyclovir treatment resulted in the termination of excretion within 8 days after onset.


Assuntos
Exantema , Infecções por Herpesviridae , Herpesvirus Equídeo 1 , Herpesvirus Equídeo 3 , Doenças dos Cavalos , Aciclovir/uso terapêutico , Animais , Exantema/veterinária , Feminino , Infecções por Herpesviridae/tratamento farmacológico , Infecções por Herpesviridae/veterinária , Doenças dos Cavalos/tratamento farmacológico , Cavalos , Japão , Masculino
7.
Virology ; 526: 105-116, 2019 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-30388626

RESUMO

Equid alpha-herpesviruses (EHV) are responsible for different diseases in equine population. EHV-1 causes respiratory diseases, abortions and nervous disorders, EHV-4 causes respiratory diseases and sporadic abortion, while EHV-3 is responsible of equine coital exanthema. In view of the lack of efficacy of vaccines against EHV-1 and EHV-4 and in the absence of vaccines against EHV-3, the use of antiviral treatment is of great interest. In this study, we documented the interest of the Real-Time Cell Analysis (RTCA) technology to monitor the cytopathic effects induced by these viruses on equine dermal cells, and established the efficacy of this method to evaluate the antiviral effect of aciclovir (ACV) and ganciclovir (GCV). In addition, the RTCA technology has also been found appropriate for the high-throughput screening of small molecules against EHV, allowing the identification of spironolactone as a novel antiviral against EHV.


Assuntos
Antivirais/farmacologia , Impedância Elétrica , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/efeitos dos fármacos , Ensaios de Triagem em Larga Escala/métodos , Animais , Linhagem Celular , Efeito Citopatogênico Viral/efeitos dos fármacos , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 3/efeitos dos fármacos , Herpesvirus Equídeo 4/efeitos dos fármacos , Cavalos , Espironolactona/farmacologia
8.
Rev Argent Microbiol ; 50(4): 380-390, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29779880

RESUMO

Equid alphaherpesvirus 3 (EHV3) is the etiological agent of equine coital exanthema (ECE), which is a venereal, highly contagious disease, characterized by the formation of papules, vesicles, pustules and ulcers on the external genitalia of mares and stallions. EHV3 remains in a latent state after a successful infection and there are latently infected animals in which the virus is reactivated and generally re-excreted subclinically. There are no available vaccines for this condition and prevention is based on the clinical examination of mares prior to mating, which allows to segregate those showing clinical signs. As this approach does not eliminate the risk of contagion in stallions from subclinically infected mares, there is a need for a specific EHV3 treatment. Nowadays, there exist various antiviral compounds of proven effectiveness for other alphaherpesviruses affecting humans and animals. The aim of the present study was to compare the efficacy of three antiviral compounds, acyclovir, ganciclovir and cidofovir against EHV3 in vitro, and to assess their efficacy against six EHV3 Argentinian field isolates. To determine the efficacy of these compounds in vitro, three parameters were analyzed: reduction of plaque number, reduction of plaque size and reduction of viral production. Additionally, the effectiveness of the three compounds at an optimum concentration previously determined in this study was investigated for the EHV3 field isolates. Based on our results, ganciclovir was the most potent antiviral compound to reduce EHV3 replication in vitro and may thus be a valuable candidate for treatment and prevention of ECE in mares and stallions.


Assuntos
Aciclovir/farmacologia , Antivirais/farmacologia , Cidofovir/farmacologia , Ganciclovir/farmacologia , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/efeitos dos fármacos , Doenças dos Cavalos/virologia , Animais , Células Cultivadas , Feminino , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 3/isolamento & purificação , Cavalos
9.
J Virol Methods ; 257: 29-32, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29627335

RESUMO

Equine coital exanthema (ECE) is an infectious, venereally transmitted muco-cutaneous disease affecting mares and stallions, caused by equid alphaherpesvirus 3 (EHV3). Diagnostic tools for rapid identification of EHV3 are of primary importance to diminish the risk of EHV3 dissemination at the time of breeding. In the last years, it has been shown that the performance of the insulated-isothermal polymerase chain reaction (iiPCR) is comparable to virus isolation, nested PCR and real-time PCR (qPCR) in detecting pathogens of various animal species. Analytical sensitivity and specificity of the iiPCR were compared with a qPCR, using a plasmid containing the target region of the EHV3 glycoprotein G gene and an Argentinian EHV3 isolate (E/9283/07 C3A). In order to evaluate the diagnostic performance of the iiPCR, nucleic acids of 85 perineal and genital swabs (PGS) of mares and stallions were extracted by tacoTM mini and tested by both techniques. EHV3 was detected in 46 and 45 of the 85 PGS by the iiPCR and qPCR, respectively. There was almost perfect agreement between the two diagnostic methods (98.82%; 95% CI: 95.03-100%; κ = 0.98). The iiPCR had a limit of detection of 95.00% at 6 genome equivalents per reaction and a detection endpoint for viral DNA comparable to that of the qPCR, and did not react with six non-targeted equine pathogens. The iiPCR represents a sensitive and specific method for the rapid on-site diagnosis of EHV3 infection. Its routinely implementation in breeding facilities, and artificial insemination and embryo transfer centers, will contribute to prevent the dissemination of this venereal, highly contagious disease in horses.


Assuntos
Genitália/virologia , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/isolamento & purificação , Doenças dos Cavalos/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Períneo/virologia , Reação em Cadeia da Polimerase/métodos , Animais , Infecções por Herpesviridae/diagnóstico , Doenças dos Cavalos/virologia , Cavalos , Testes Imediatos , Sensibilidade e Especificidade
10.
Theriogenology ; 109: 22-30, 2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29249328

RESUMO

This paper reviews that state of our knowledge concerning follicular wave dynamics, monitoring and manipulation. All camelids have overlapping follicular waves in absence of ovulation which is induced by a seminal plasma factor (ßNGF). The interval between follicular waves varies. The size of the ovulatory follicle varies between 11 and 25 mm in camels and between in 6 and 13 mm in South American Camelids. The interval between induction of ovulation and next ovulatory follicle is 15 ± 1 day for all camelids. Follicular activity is best monitored by transrectal ultrasonography. Progesterone therapy for 7-15 days seems to suppress follicular dominance but does not completely inhibit follicular recruitment. Combination of estradiol and progesterone seems to provide better control of follicular activity. Both methods have provided variable results in the synchronization of follicular waves. Combination of induction of ovulation with GnRH and luteolysis at predetermined times shows some promise in synchronization of follicular dominance. These synchronization protocols require further investigation in order to provide practical approaches for fixed-time breeding. Ovarian superstimulation with FSH and eCG alone or in combination is somewhat successful. The best results are obtained when treatment is initiated at the emergence of a new follicular wave after induction of ovulation or following treatment with progesterone for 7-14 days. However, response remains extremely variable particularly in terms of ovulation rate and number of recovered embryos. Sources of this variability need to be studied including the effects of season, nutrition, doses and frequency of administration of gonadotropin.


Assuntos
Camelidae/fisiologia , Folículo Ovariano/fisiologia , Animais , Cruzamento/métodos , Estradiol/administração & dosagem , Sincronização do Estro/métodos , Feminino , Hormônio Foliculoestimulante/administração & dosagem , Hormônio Liberador de Gonadotropina/farmacologia , Gonadotropinas Equinas/administração & dosagem , Herpesvirus Equídeo 3 , Luteólise/fisiologia , Monitorização Fisiológica/métodos , Monitorização Fisiológica/veterinária , Folículo Ovariano/anatomia & histologia , Folículo Ovariano/efeitos dos fármacos , Ovário/fisiologia , Ovulação/fisiologia , Indução da Ovulação/métodos , Indução da Ovulação/veterinária , Progesterona/administração & dosagem , Ultrassonografia/métodos , Ultrassonografia/veterinária
11.
J Vet Med Sci ; 79(3): 632-635, 2017 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-28123140

RESUMO

Equine coital exanthema (ECE) has been reported in many countries, but equine herpesvirus 3 (EHV-3) has been isolated only once in Japan. In 2015, symptoms of ECE were found, and EHV-3 was isolated in two stallions. Valacyclovir, an anti-herpesvirus agent, was administered orally. The stallions rested from mating for more than two weeks, causing enormous financial losses because of their high fees. This is the first study in which valacyclovir was administered for ECE. Though valacyclovir treatment did not shorten the duration of healing, the affected area did not expand after administration of valacyclovir. Valacyclovir therefore seems to be effective for suppression of EHV-3 infection. Further investigation about the administration protocol might be required.


Assuntos
Aciclovir/análogos & derivados , Exantema/veterinária , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/isolamento & purificação , Doenças dos Cavalos/epidemiologia , Doenças Virais Sexualmente Transmissíveis/veterinária , Valina/análogos & derivados , Aciclovir/uso terapêutico , Animais , Antivirais/uso terapêutico , Exantema/tratamento farmacológico , Exantema/epidemiologia , Exantema/virologia , Infecções por Herpesviridae/tratamento farmacológico , Infecções por Herpesviridae/epidemiologia , Doenças dos Cavalos/tratamento farmacológico , Doenças dos Cavalos/virologia , Cavalos , Japão , Masculino , Doenças Virais Sexualmente Transmissíveis/tratamento farmacológico , Doenças Virais Sexualmente Transmissíveis/epidemiologia , Valaciclovir , Valina/uso terapêutico
12.
J Vet Med Sci ; 79(3): 636-643, 2017 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-28132964

RESUMO

In the spring of 2015, two stallions reared in Farms A and B in Hokkaido in Japan showed symptoms of equine coital exanthema. Equine herpesvirus 3 (EHV-3) was isolated from penis swab samples of both stallions, and the isolates from each stallion in Farms A and B were designated as SS-1 and YS-1 strains, respectively. BamHI restriction profiles of SS-1 and Japanese reference strain Iwate-1 were indistinguishable, but the BamHI-A fragment of YS-1 was larger than those of SS-1 and Iwate-1 by 1.9 kbp because of the lack of two BamHI sites. Nucleotide sequence analyses of glycoprotein G (gG), gB, gC and VP13/14 coding regions revealed that SS-1 and YS-1 had 99.77% to 100% identities to each other. These results suggested that the origins of SS-1 and YS-1 were different. For a sero-epidemiological survey, serum neutralizing tests using SS-1 against 319 sera of horses from eight farms in Hokkaido were conducted. Six of the eight farms were EHV-3 antibody-positive, and positive rates ranged from 2.6% to 17.6%. To determine the infection time of four EHV-3 antibody-positive horses, a retrospective study was conducted. Infection time of the four horses was in the breeding season, and re-infection or reactivation of latently infected EHV-3 might have occurred in one horse. However, these four horses had never shown any clinical symptoms. The results suggested that several EHV-3 strains are distributed in Japan and that infection is maintained widely in horses without clinical symptoms.


Assuntos
Exantema/veterinária , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/isolamento & purificação , Doenças dos Cavalos/virologia , Doenças Virais Sexualmente Transmissíveis/veterinária , Animais , Exantema/epidemiologia , Exantema/virologia , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Doenças dos Cavalos/epidemiologia , Cavalos , Japão , Masculino , Análise de Sequência de DNA , Estudos Soroepidemiológicos , Doenças Virais Sexualmente Transmissíveis/epidemiologia , Doenças Virais Sexualmente Transmissíveis/virologia
13.
Virus Res ; 228: 30-38, 2017 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-27865864

RESUMO

Equine herpesvirus type 3 (EHV-3) is the causal agent of equine coital exanthema, a disease characterized by pox-like lesions on the penis of stallions and the vulva of mares. Although the complete genomic sequence of EHV-3 has been recently made available, its genomic content remains poorly characterized and the molecular mechanisms of disease development not yet elucidated. In an attempt to facilitate genetic manipulation of EHV-3, we describe here the construction of a full-length infectious bacterial artificial chromosome (BAC) clone of EHV-3. Mini-F vector sequences were inserted into the intergenic region between ORF19 and ORF20 (UL41 and UL40, respectively) of EHV-3 strain C175 by homologous recombination in equine dermal cells (NBL-6). DNA of the resulting recombinant virus was electroporated into E. coli and a full-length EHV-3 BAC clone was recovered. Virus reconstituted after transfection of the EHV-3 BAC into NBL-6 cells showed growth properties in vitro that were indistinguishable from those of the parental virus. To assess the feasibility of mutagenesis of the cloned EHV-3 genome, recombinant viruses targeting the glycoprotein E (gE) gene were generated using Red recombination in E. coli and in vitro growth properties of the recombinant viruses were evaluated. We first repaired the gE (ORF74) coding region, since the parental virus used for BAC cloning specifies a truncated version of the gene, and then created gE-tagged and gE-null versions of the virus. Our results demonstrated that: (i) EHV-3 can be efficiently cloned as a BAC allowing easy manipulation of its genome; (ii) gE is dispensable for EHV-3 growth in vitro and is expressed as a product of approximately 110-kDa in infected cells; (iii) viruses having a deletion compromising gE expression or with a truncation of the cytoplasmic and transmembrane domains are significantly compromised with regard cell-to-cell spread. The cloning of EHV-3 as a BAC simplifies future studies to identify the role of its coding genes in viral pathogenesis and host immune responses.


Assuntos
Cromossomos Artificiais Bacterianos , DNA Recombinante , Vetores Genéticos , Genoma Viral , Herpesvirus Equídeo 3/genética , Células Cultivadas , Clonagem Molecular , Expressão Gênica , Ordem dos Genes , Engenharia Genética , Vetores Genéticos/genética , Mutagênese , Fases de Leitura Aberta , Transfecção , Ensaio de Placa Viral , Replicação Viral
14.
Vet Res ; 47: 19, 2016 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-26768993

RESUMO

Replication kinetics and invasion characteristics of equine herpesvirus-1 and -3 (EHV-1/-3) in nasal and vaginal mucosae were compared using explants. The explants were cultured during 96 h with little change in viability. The tissues were inoculated with EHV-1 03P37 (neuropathogenic), 97P70 (abortigenic) and EHV-3 04P57, collected at 0, 24, 48 and 72 h post inoculation (pi) and stained for viral antigens. Both EHV-1 and EHV-3 replicated in a plaquewise manner. The plaques were already observed at 24 h pi, their size increased over time and did not directly cross the basement membrane (BM). However, EHV-1 infected the monocytic cells (MC) and hijacked these cells to invade the lamina propria. In contrast, EHV-3 replication was fully restricted to epithelial cells; the virus did not breach the BM via a direct cell-to-cell spread nor used infected MC. EHV-1-induced plaques were larger in nasal mucosa compared to vaginal mucosa. The opposite was found for EHV-3-induced plaques. Both EHV-1 strains replicated with comparable kinetics in nasal mucosa. However, the extent of replication of the abortigenic strain in vaginal mucosa was significantly higher than that of the neuropathogenic strain. Two-to-five-fold lower numbers of EHV-1-infected MC underneath the BM were found in vaginal mucosa than in nasal mucosa. Our study has shown that (i) EHV-1 has developed in evolution a predisposition for respiratory mucosa and EHV-3 for vaginal mucosa, (ii) abortigenic EHV-1 replicates better in vaginal mucosa than neuropathogenic EHV-1 and (iii) EHV-3 demonstrated a strict epithelial tropism whereas EHV-1 in addition hijacked MC to invade the lamina propria.


Assuntos
Herpesvirus Equídeo 1/fisiologia , Herpesvirus Equídeo 3/fisiologia , Mucosa/virologia , Replicação Viral/fisiologia , Animais , Feminino , Cavalos , Mucosa/citologia , Nariz , Técnicas de Cultura de Tecidos , Vagina
15.
Vet Ophthalmol ; 18(5): 416-21, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25594353

RESUMO

OBJECTIVES: To determine the role of equine herpesvirus (EHV) in idiopathic keratoconjunctivitis in horses and to determine whether sample collection method affects detection of EHV DNA by quantitative polymerase chain reaction (qPCR). ANIMALS STUDIED: Twelve horses with idiopathic keratoconjunctivitis and six horses without signs of ophthalmic disease. PROCEDURES: Conjunctival swabs, corneal scrapings, and conjunctival biopsies were collected from 18 horses: 12 clinical cases with idiopathic keratoconjunctivitis and six euthanized controls. In horses with both eyes involved, the samples were taken from the eye judged to be more severely affected. Samples were tested with qPCR for EHV-1, EHV-2, EHV-4, and EHV-5 DNA. Quantity of EHV DNA and viral replicative activity were compared between the two populations and among the different sampling techniques; relative sensitivities of the sampling techniques were determined. RESULTS: Prevalence of EHV DNA as assessed by qPCR did not differ significantly between control horses and those with idiopathic keratoconjunctivitis. Sampling by conjunctival swab was more likely to yield viral DNA as assessed by qPCR than was conjunctival biopsy. EHV-1 and EHV-4 DNA were not detected in either normal or IKC-affected horses; EHV-2 DNA was detected in two of 12 affected horses but not in normal horses. EHV-5 DNA was commonly found in ophthalmically normal horses and horses with idiopathic keratoconjunctivitis. CONCLUSIONS: Because EHV-5 DNA was commonly found in control horses and in horses with idiopathic keratoconjunctivitis, qPCR was not useful for the etiological diagnosis of equine keratoconjunctivitis. Conjunctival swabs were significantly better at obtaining viral DNA samples than conjunctival biopsy in horses in which EHV-5 DNA was found.


Assuntos
Infecções por Herpesviridae/veterinária , Doenças dos Cavalos/diagnóstico , Ceratoconjuntivite Infecciosa/diagnóstico , Animais , Feminino , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 1 , Herpesvirus Equídeo 3 , Herpesvirus Equídeo 4 , Doenças dos Cavalos/virologia , Cavalos , Ceratoconjuntivite Infecciosa/virologia , Masculino , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Rhadinovirus , Manejo de Espécimes/veterinária , Varicellovirus , Carga Viral/veterinária
16.
Vet Microbiol ; 160(3-4): 319-26, 2012 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-22776511

RESUMO

Equine coital exanthema (ECE), caused by equid herpesvirus 3 (EHV-3), has been recognized as an economically significant venereal disease for years. However, no infection models on the natural host have been established. In order to set up an experimental infection protocol, seronegative and seropositive mares were topically inoculated in the perineal region with 4 × 10(6)TCID(50)/ml of EHV-3. Clinical signs were then evaluated by means of a designed scoring system, and body temperature was recorded daily. Virological, and serological studies were also performed. Typical ECE lesions, with clinical scores of 90, 92, 160 and 172, were observed in the four seronegative animals. Only mild ECE lesions were observed in the two seropositive mares, being the clinical scores 53 and 41. Both groups of mares shed the virus, but the duration of virus shedding was shorter and its intensity was lower in seropositive mares than in seronegative ones. Moreover, EHV-3 antibody response was detected in both seronegative and seropositive mares after experimental infection and re-infection, being more moderate in seropositive ones. As a conclusion, EHV-3 infection of mares was experimentally achieved in a reproducible manner. The typical lesions of ECE were observed after topical EHV-3 infection in seronegative mares, in association with virus excretion and neutralizing antibody kinetics.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/fisiologia , Doenças dos Cavalos/imunologia , Doenças dos Cavalos/patologia , Infecções Sexualmente Transmissíveis/veterinária , Animais , Anticorpos Antivirais/sangue , Formação de Anticorpos , Sequência de Bases , Feminino , Genótipo , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologia , Doenças dos Cavalos/virologia , Cavalos , Dados de Sequência Molecular , Alinhamento de Sequência , Infecções Sexualmente Transmissíveis/imunologia , Infecções Sexualmente Transmissíveis/patologia , Infecções Sexualmente Transmissíveis/virologia , Fatores de Tempo , Proteínas do Envelope Viral/genética , Eliminação de Partículas Virais
17.
Vet J ; 191(1): 35-40, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21354835

RESUMO

Equine coital exanthema (ECE) caused by equid herpesvirus 3 (EHV-3) is a contagious venereal disease characterised by the formation of painful papules, vesicles, pustules and ulcers on the external genitalia of both mares and stallions. EHV-3 is an alphaherpesvirus that is distinct from the other equine herpesviruses and endemic in most horse breeding populations worldwide. The negative impacts of ECE on equine breeding enterprises are the forced, temporary disruption of mating activities of mares and stallions, the additional care and supportive treatment that is required for affected horses, and the risk of virus spread by either fresh or frozen semen as well as by artificial insemination and embryo transfer. Because there are no effective surveillance systems to report ECE, its true prevalence and economic impact are difficult to assess and are probably underestimated. The purpose of this review is to describe the recent advances in understanding of EHV-3 infections and to consider the economic consequences of ECE within the current context of the equine industry.


Assuntos
Exantema/veterinária , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/isolamento & purificação , Infecções Sexualmente Transmissíveis/veterinária , Animais , Bélgica , Cruzamento , Transmissão de Doença Infecciosa/veterinária , Transferência Embrionária/veterinária , Exantema/prevenção & controle , Feminino , Genitália/virologia , Infecções por Herpesviridae/prevenção & controle , Cavalos , Indústrias , Inseminação Artificial/veterinária , Masculino , Infecções Sexualmente Transmissíveis/prevenção & controle
18.
Theriogenology ; 74(4): 576-80, 2010 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-20494427

RESUMO

The temporary disruption of reproductive activities due to equine coital exanthema (ECE), caused by equid herpesvirus 3 (EHV-3), at thoroughbred breeding facilities and embryo transfer centres, has an appreciable economic impact. The aim of the present study was to estimate the prevalence of excretion of EHV-3 in mares without clinical symptoms under field conditions and the re-excretion patterns of the virus in two seropositive (presumably latently infected) mares maintained in isolation for 11 mo. The EHV-3 virus was detected in perineal-vaginal swabs by real time PCR in 14 (6%) of 220 thoroughbred mares without clinical symptoms at the time of breeding. In the two isolated mares, re-excretion of EHV-3 was demonstrated on two occasions, 3 mo apart (each for a 3 d interval) in one mare, and on only 1 d in the other mare. Antibodies against EHV-3 were identified by seroneutralization in 105 (48%) of the thoroughbred mares, and during the entire period in the two isolated mares. Therefore, the present study provided evidence of EHV-3 shedders in a healthy mare population under both field and isolation conditions. Furthermore, at least two periods of spontaneous EHV-3 reactivation and re-excretion in the presence of serum antibodies occurred in one mare in an 11 mo interval. These findings could assist in the design and implementation of measures to minimize the spread of EHV-3 and control ECE outbreaks.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 3/isolamento & purificação , Doenças dos Cavalos/virologia , Eliminação de Partículas Virais , Animais , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 3/fisiologia , Doenças dos Cavalos/imunologia , Cavalos , Periodicidade , Ativação Viral
20.
Vet Immunol Immunopathol ; 135(1-2): 93-99, 2010 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-20004981

RESUMO

Recombinant equine interferon-gamma (reIFN-gamma) was prepared using a baculovirus expression system and its antiviral activity was investigated using several equine viruses. The reIFN-gamma suppressed the replication of all equine viruses used in the present experiment in horse cell cultures, but did not affect the growth of host cells at concentrations of less than 1000 u/ml. A strong antiviral effect was observed, especially against RNA viruses. Equine picornavirus, equine rhinovirus and equine arteritis virus could not be propagated at all in 100 u/ml reIFN-gamma when 100 TCID(50) of infective viruses was inoculated to cultivated horse cells. DNA viruses, equine herpesvirus types 1, 2, 3 and 4 and equine adenovirus, were less sensitive to reIFN-gamma but their growth became less than 1/100 in the cells treated with 100 u/ml reIFN-gamma compared to untreated cells. The antiviral effects were decreased in the cells of heterologous species and more than 1000 u/ml reIFN-gamma was required to induce an antiviral effect.


Assuntos
Adenoviridae/efeitos dos fármacos , Alphavirus/efeitos dos fármacos , Antivirais/farmacologia , Aphthovirus/efeitos dos fármacos , Equartevirus/efeitos dos fármacos , Herpesvirus Equídeo 1/efeitos dos fármacos , Herpesvirus Equídeo 3/efeitos dos fármacos , Herpesvirus Equídeo 4/efeitos dos fármacos , Interferon gama/farmacologia , Picornaviridae/efeitos dos fármacos , Rhadinovirus/efeitos dos fármacos , Animais , Antivirais/efeitos adversos , Células Cultivadas , Relação Dose-Resposta a Droga , Doenças dos Cavalos/tratamento farmacológico , Doenças dos Cavalos/virologia , Cavalos/virologia , Interferon gama/efeitos adversos , Proteínas Recombinantes , Replicação Viral/efeitos dos fármacos
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