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2.
Arch Virol ; 164(12): 2975-2984, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31570994

RESUMO

Coxsackievirus A16 (CV-A16) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children and has become prevalent in the Asia-Pacific region in recent years. However, no approved vaccines or drugs are available for CV-A16 infection. CV-A16 virus-like particles (VLPs) are a potential vaccine candidate; however, whether the intranasal route of immunization is suitable for inducing immune responses against CV-A16 infection has not been clarified. In this study, the comprehensive immunogenicity and protective efficacy of the CV-A16 VLP vaccine were evaluated by multiple methods in a mouse model. In mice, a high neutralizing antibody (NTAb) titre could be elicited by intranasal immunization with CV-A16 VLPs, which produced NTAb levels similar to those induced by intranasal immunization with inactivated CV-A16. Passive immunity with NTAbs provided very good protection, as the survival rate of the immunized neonatal mice was 100% after challenges with CV-A16 at a dose of 1000 LD50. Passive protective effects were transferred to the neonates via the mother, thus protecting all the pups against challenges with the homologous or heterologous strains of CV-A16 at a dose of 1000 LD50. In addition, intranasal immunization with CV-A16 VLPs also induced the production of mucosal secretory IgA (s-IgA) antibodies, which may inhibit CV-A16 virus invasion. This study provides valuable supplemental information to facilitate our understanding of the specific protective efficacy of CV-A16 VLPs and has significance for development of the candidate vaccine into a safe and effective vaccine.


Assuntos
Enterovirus Humano A/imunologia , Infecções por Enterovirus/prevenção & controle , Nariz/virologia , Vacinas Virais/administração & dosagem , Animais , Animais Recém-Nascidos , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Modelos Animais de Doenças , Enterovirus Humano A/genética , Infecções por Enterovirus/imunologia , Infecções por Enterovirus/virologia , Feminino , Humanos , Imunização , Camundongos , Camundongos Endogâmicos ICR , Vacinas Virais/genética , Vacinas Virais/imunologia
3.
Food Environ Virol ; 11(4): 340-349, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31350695

RESUMO

Rhinovirus is a common picornavirus with over 150 serotypes and three species, which is responsible for half of the human common cold cases. In people with chronic respiratory conditions and elders, it may also cause life-threatening diseases. Transmission routes are not definitively established but may involve direct human-to-human and indirect transmission (surfaces and aerosols based). In the present study, year-long presence of virus was tested by qPCR in the nostrils of young healthy volunteers and indoor and outdoor air samples. Results were correlated to atmospheric conditions (meteorological and air quality parameters) and voluntaries immune system-related genetic polymorphisms (TOLLIP rs5743899, IL6 rs1800795, IL1B rs16944, TNFA rs1800629) typed by PCR-RFLP. Nasal samples showed increased frequency and viral titers of Rhinovirus in spring and autumn. No indoor air samples tested positive for Rhinovirus, whereas outdoor air samples tested positive in late autumn. Sun radiation, atmospheric SO2, and benzene levels correlated with nostrils Rhinovirus detection. Both IL6 and TOLLIP polymorphisms but not TNFA or IL1B influenced Rhinovirus detection in the nostrils of voluntaries. Taken together, the results indicate that Rhinovirus circulation is determined by environmental conditions (weather, air-borne virus, and air pollution) and genetically encoded individual variation in immunity.


Assuntos
Infecções por Picornaviridae/genética , Infecções por Picornaviridae/imunologia , Polimorfismo Genético , Rhinovirus/fisiologia , Adulto , Ar/análise , Microbiologia do Ar , Poluição do Ar , Feminino , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , Masculino , Nariz/imunologia , Nariz/virologia , Infecções por Picornaviridae/virologia , Rhinovirus/genética , Rhinovirus/isolamento & purificação , Estações do Ano , Adulto Jovem
4.
Virol J ; 16(1): 77, 2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31174549

RESUMO

BACKGROUND: Influenza B virus is a main causative pathogen of annual influenza epidemics, however, research on influenza B virus in general lags behind that on influenza A viruses, one of the important reasons is studies on influenza B viruses in animal models are limited. Here we investigated the tree shrew as a potential model for influenza B virus studies. METHODS: Tree shrews and ferrets were inoculated with either a Yamagata or Victoria lineage influenza B virus. Symptoms including nasal discharge and weight loss were observed. Nasal wash and respiratory tissues were collected at 2, 4 and 6 days post inoculation (DPI). Viral titers were measured in nasal washes and tissues were used for pathological examination and extraction of mRNA for measurement of cytokine expression. RESULTS: Clinical signs and pathological changes were also evident in the respiratory tracts of tree shrews and ferrets. Although nasal symptoms including sneezing and rhinorrhea were evident in ferrets infected with influenza B virus, tree shrews showed no significant respiratory symptoms, only milder nasal secretions appeared. Weight loss was observed in tree shrews but not ferrets. V0215 and Y12 replicated in all three animal (ferrets, tree shrews and mice) models with peak titers evident on 2DPI. There were no significant differences in peak viral titers in ferrets and tree shrews inoculated with Y12 at 2 and 4DPI, but viral titers were detected at 6DPI in tree shrews. Tree shrews infected with influenza B virus showed similar seroconversion and respiratory tract pathology to ferrets. Elevated levels of cytokines were detected in the tissues isolated from the respiratory tract after infection with either V0215 or Y12 compared to the levels in the uninfected control in both animals. Overall, the tree shrew was sensitive to infection and disease by influenza B virus. CONCLUSION: The tree shrew to be a promising model for influenza B virus research.


Assuntos
Anticorpos Antivirais/sangue , Modelos Animais de Doenças , Vírus da Influenza B/imunologia , Infecções por Orthomyxoviridae/imunologia , Tupaiidae/virologia , Animais , Citocinas/imunologia , Feminino , Furões , Vírus da Influenza B/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nariz/virologia , Sistema Respiratório/imunologia , Sistema Respiratório/virologia , Árvores , Carga Viral , Replicação Viral
5.
Braz J Microbiol ; 50(3): 875-878, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31187445

RESUMO

Equid gammaherpesvirus 2 (EHV-2) and 5 (EHV-5) are members of the Herpesviridae family and have been reported in horse populations worldwide. This study aimed to evaluate the presence of herpesvirus DNA in the upper respiratory tract of horses. Twenty-six nasal swabs were collected from asymptomatic adult horses of two different horse farms (A, n = 18; B, n = 8), both located in Southern Brazil. The EHV-1, EHV-2, EHV-4, and EHV-5 DNA analyses were performed using nested PCR assays targeting the glycoprotein B gene. Four (15.3%) and 12 (46.1%) of the 26 nasal swab samples were positive for the EHV-2 and EHV-5, respectively. Four (15.3%) horses were detected with both viruses simultaneously. DNA of EHV-2 and EHV-5 in both single and mixed infections was identified in horses from both herds. All swab samples were negative for EHV-1 and EHV-4. This study reports the first detection of EHV-2 and EHV-5 in the upper respiratory tracts of horses in Brazil. The high detection rate of EHV-2 and EHV-5 in asymptomatic adult horses demonstrates that these gammaherpesviruses are circulating in Brazil.


Assuntos
Gammaherpesvirinae/isolamento & purificação , Infecções por Herpesviridae/veterinária , Doenças dos Cavalos/virologia , Nariz/virologia , Animais , Doenças Assintomáticas , Brasil , DNA Viral/genética , Feminino , Gammaherpesvirinae/classificação , Gammaherpesvirinae/genética , Infecções por Herpesviridae/virologia , Cavalos , Masculino
6.
Viruses ; 11(4)2019 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-31022948

RESUMO

Middle East respiratory syndrome coronavirus (MERS-CoV) transmission from dromedaries to humans has resulted in major outbreaks in the Middle East. Although some other livestock animal species have been shown to be susceptible to MERS-CoV, it is not fully understood why the spread of the virus in these animal species has not been observed in the field. In this study, we used rabbits to further characterize the transmission potential of MERS-CoV. In line with the presence of MERS-CoV receptor in the rabbit nasal epithelium, high levels of viral RNA were shed from the nose following virus inoculation. However, unlike MERS-CoV-infected dromedaries, these rabbits did not develop clinical manifestations including nasal discharge and did shed only limited amounts of infectious virus from the nose. Consistently, no transmission by contact or airborne routes was observed in rabbits. Our data indicate that despite relatively high viral RNA levels produced, low levels of infectious virus are excreted in the upper respiratory tract of rabbits as compared to dromedary camels, thus resulting in a lack of viral transmission.


Assuntos
Infecções por Coronavirus/transmissão , Coronavírus da Síndrome Respiratória do Oriente Médio/fisiologia , Nariz/virologia , Coelhos/virologia , Organismos Livres de Patógenos Específicos , Animais , Anticorpos Antivirais/sangue , Camelus/virologia , Infecções por Coronavirus/virologia , Reservatórios de Doenças/virologia , Feminino , Masculino , Coronavírus da Síndrome Respiratória do Oriente Médio/imunologia , RNA Viral/análise , Sistema Respiratório/virologia , Eliminação de Partículas Virais
7.
Viruses ; 11(3)2019 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-30871054

RESUMO

Following the successful eradication of rinderpest, the World Organization of Animal Health (OIE) and the Food and Agriculture Organisation (FAO) have set a goal to globally eradicate Peste des petits ruminants (PPR) by 2030. To support the eradication programme we have quantified the levels of PPR virus (PPRV) nucleic acid excreted in body fluids (blood, feces, saliva, nasal and eye swabs) of PPRV-infected goats to ascertain which days post-infection animals are potentially infectious, and hence direct quarantine activities. The data will also indicate optimal sample strategies to assess presence of PPR infection in the naturally infected herd. Peak PPRV nucleic acid detection in different bodily fluids was between 5 and 10 days post-infection. As such, this period must be considered the most infectious period for contact transmission, although high viral load was observed through RNA detection in nasal excretions from two days post-infection until at least two weeks post-infection. Percentage sample positivity was low both in eye swabs and saliva samples during the early stage of infection although RNA was detected as late as two weeks post-infection. From the individual animal data, PPRV was detected later post-infection in fecal material than in other body fluids and the detection was intermittent. The results from this study indicate that nasal swabs are the most appropriate to sample when considering molecular diagnosis of PPRV.


Assuntos
Erradicação de Doenças , Surtos de Doenças/veterinária , Doenças das Cabras/virologia , Peste dos Pequenos Ruminantes/prevenção & controle , Vírus da Peste dos Pequenos Ruminantes/isolamento & purificação , RNA Viral/análise , RNA Viral/genética , Animais , Surtos de Doenças/prevenção & controle , Fezes/virologia , Cabras , Masculino , Nariz/virologia , Peste dos Pequenos Ruminantes/sangue , Vírus da Peste dos Pequenos Ruminantes/genética , Reação em Cadeia da Polimerase em Tempo Real , Saliva/virologia
8.
Emerg Microbes Infect ; 8(1): 94-102, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30866763

RESUMO

There was a substantial increase with infections of H7N9 avian influenza virus (AIV) in humans during Wave 5 (2016-2017). To investigate whether H7N9 had become more infectious/transmissible and pathogenic overall, we characterized the receptor binding and experimentally infected ferrets with highly pathogenic (HP)- and low pathogenic (LP)-H7N9 isolates selected from Wave 5, and compared their pathogenicity and transmissibility with a Wave 1 isolate from 2013. Studies show that A/Anhui/1/2013 (LP) and A/Chicken/Heyuan/16876/2016 (HP) were highly virulent in ferrets, A/Guangdong/Th008/2017 (HP) and A/Chicken/Huizhou/HZ-3/2017 (HP) had moderate virulence and A/Shenzhen/Th001/2016 (LP) was of low virulence in ferrets. Transmission was observed only in ferrets infected with A/Anhui/1/2013 and A/Chicken/Heyuan/16876/2016, consistent with the idea that sicker ferrets had a higher probability to transmit virus to naive animals. Given the Varied virulence and transmissibility observed in circulating H7N9 viruses from Wave 5, we conclude that the current public health risk of H7N9 has not substantially increased compared to 2013 and the circulating viruses are quite diverse.


Assuntos
Furões/virologia , Subtipo H7N9 do Vírus da Influenza A/patogenicidade , Infecções por Orthomyxoviridae/transmissão , Receptores de Superfície Celular/metabolismo , Proteínas Virais/metabolismo , Animais , Genótipo , Humanos , Subtipo H7N9 do Vírus da Influenza A/genética , Subtipo H7N9 do Vírus da Influenza A/isolamento & purificação , Subtipo H7N9 do Vírus da Influenza A/metabolismo , Influenza Humana/virologia , Nariz/virologia , Infecções por Orthomyxoviridae/virologia , Faringe/virologia , Virulência
9.
Emerg Microbes Infect ; 8(1): 103-108, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30866764

RESUMO

Dromedary camels are natural host of the Middle East respiratory syndrome coronavirus (MERS-CoV). However, there are limited studies of MERS-CoV infection of other domestic mammals exposed to infected dromedaries. We expanded our surveillance among camels in Egypt, Tunisia, and Senegal to include other domestic mammalian species in contact with infected camels. A total of 820 sera and 823 nasal swabs from cattle, sheep, goats, donkeys, buffaloes, mules, and horses were collected. Swabs were tested using RT-PCR and virus RNA-positive samples were genetically sequenced and phylogenetically analysed. Sera were screened using virus microneutralization tests and positive sera (where available) were confirmed using plaque reduction neutralization tests (PRNT). We detected 90% PRNT confirmed MERS-CoV antibody in 35 (55.6%) of 63 sera from sheep collected from Senegal, two sheep (1.8%) of 114 in Tunisia and a goat (0.9%) of 107 in Egypt, with titres ranging from 1:80 to ≥1:320. We detected MERS-CoV RNA in swabs from three sheep (1.2%) of 254 and five goats (4.1%) of 121 from Egypt and Senegal, as well as one cow (1.9%) of 53 and three donkeys (7.1%) of 42 from Egypt. Partial sequences of the RT-PCR amplicons confirmed specificity of the results. This study showed that domestic livestock in contact with MERS-CoV infected camels may be at risk of infection. We recommend expanding current MERS-CoV surveillance in animals to include other livestock in close contact with dromedary camels. The segregation of camels from other livestock in farms and live animal markets may need to be considered.


Assuntos
Animais Domésticos/virologia , Anticorpos Antivirais/sangue , Infecções por Coronavirus/diagnóstico , Coronavírus da Síndrome Respiratória do Oriente Médio/isolamento & purificação , Nariz/virologia , Análise de Sequência de RNA/métodos , Animais , Animais Domésticos/sangue , Animais Domésticos/imunologia , Bovinos , Infecções por Coronavirus/imunologia , Egito , Cabras , Cavalos , Humanos , Coronavírus da Síndrome Respiratória do Oriente Médio/classificação , Coronavírus da Síndrome Respiratória do Oriente Médio/genética , Coronavírus da Síndrome Respiratória do Oriente Médio/imunologia , Testes de Neutralização , Filogenia , Vigilância da População , Senegal , Ovinos , Tunísia
10.
Influenza Other Respir Viruses ; 13(4): 319-330, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30925029

RESUMO

BACKGROUND: Internet-based participatory surveillance systems, such as the German GrippeWeb, monitor the frequency of acute respiratory illnesses on population level. In order to interpret syndromic information better, we devised a microbiological feasibility study (GrippeWeb-Plus) to test whether self-collection of anterior nasal swabs is operationally possible, acceptable for participants and can yield valid data. METHODS: We recruited 103 GrippeWeb participants (73 adults and 30 children) and provided them with a kit, instructions and a questionnaire for each sample. In the first half of 2016, participants took an anterior nasal swab and sent it to the Robert Koch Institute whenever an acute respiratory illness occurred. Reporting of illnesses through the GrippeWeb platform continued as usual. We analysed swabs for the presence of human c-myc-DNA and 22 viral and bacterial pathogens. After the study, we sent participants an evaluation questionnaire. We analysed timeliness, completeness, acceptability and validity. RESULTS: One hundred and two participants submitted 225 analysable swabs. Ninety per cent of swabs were taken within 3 days of symptom onset. Eighty-nine per cent of swabs had a corresponding reported illness in the GrippeWeb system. Ninety-nine per cent of adults and 96% of children would be willing to participate in a self-swabbing scheme for a longer period. All swabs contained c-myc-DNA. In 119 swabs, we identified any of 14 viruses but no bacteria. The positivity rate of influenza was similar to that in the German physician sentinel. CONCLUSION: Self-collection of anterior nasal swabs proofed to be feasible, was well accepted by participants, gave valid results and was an informative adjunct to syndromic data.


Assuntos
Monitoramento Epidemiológico , Nariz/virologia , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Manejo de Espécimes/métodos , Vírus/isolamento & purificação , Doença Aguda/epidemiologia , Adulto , Criança , Estudos de Viabilidade , Alemanha , Humanos , Influenza Humana/diagnóstico , Manejo de Espécimes/normas , Inquéritos e Questionários , Vírus/genética
11.
Influenza Other Respir Viruses ; 13(1): 91-105, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29996007

RESUMO

BACKGROUND: Influenza B viruses (IBVs) have never been isolated from natural-infected pigs in clinical cases, although the susceptibility of domestic pigs to experimental IBV infections had been confirmed as well as IBV-specific antibodies were detected from pigs under natural and experimental conditions. OBJECTIVES: We aimed to assess and investigate the activities for infection and circulation of IBVs in pigs. METHODS: Annual active surveys for influenza have been implemented on swine populations in Taiwan since July 1998. Nasal swabs, trachea, lungs, and blood from pigs were tested using virological and serological assays for influenza. Gene sequences of influenza viral isolates were determined and characterized. Preliminary sero-epidemiological data for influenza virus were investigated. RESULTS: Three strains of IBV were isolated and identified from natural-infected pigs in 2014. Genetic characterization revealed the highest identities (>99%) of molecular sequence with the contemporary IBVs belonged to the B/Brisbane/60/2008 genetic clade of Victoria lineage in the phylogenetic trees for all 8 genes. IBV-specific antibodies were detected in 31 (0.2%; 95%CI: 0.1%-0.2%) of 15 983 swine serum samples from 29 (2.8%; 95%CI: 1.9%-3.9%) of 1039 farm visits under annual active surveys from 2007 through 2017. Seropositive cases have been found sparsely in 1-5 of test prefectures every year except 2015 and 2017 as well as scattered loosely over 26 townships/districts of 11 prefectures in Taiwan cumulatively in 11 years. CONCLUSIONS: Influenza B viruse infections from humans to pigs remained sporadic and accidental currently in Taiwan but might have paved potential avenues for newly emerging zoonotic influenza in the future.


Assuntos
Anticorpos Antivirais/sangue , Vírus da Influenza B/isolamento & purificação , Infecções por Orthomyxoviridae/veterinária , Doenças dos Suínos/epidemiologia , Animais , Fazendas , Vírus da Influenza B/genética , Nariz/virologia , Infecções por Orthomyxoviridae/epidemiologia , Filogenia , Testes Sorológicos/veterinária , Sus scrofa , Suínos , Doenças dos Suínos/virologia , Taiwan/epidemiologia , Replicação Viral
12.
Ophthalmic Plast Reconstr Surg ; 35(1): e21-e22, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30562334

RESUMO

This is the first reported case of necrotizing nasal and sinus herpes infection with orbital involvement. A 94-year-old male with a past medical history of hypertension, congestive heart failure, and chronic kidney disease presented with necrotic nasal cellulitis and a red left eye with blurry vision. Onset was 2 weeks prior with an upper lip fever blister. He was admitted to medicine and started on antimicrobials. Maxillofacial computed tomography scan revealed pansinusitis with involvement of bilateral medial orbital walls. Initial nasal biopsy was negative. Repeat biopsy was positive for herpes simplex virus 1. His nose was debrided with full-thickness skin grafting. While there are reports of necrotizing herpes simplex, none were to the extent of our patient nor raised suspicion for invasive fungal sinusitis. Differentiating is important as this patient made a full recovery in comparison to patients with mucormycosis where mortality is often as high as 80%.


Assuntos
Herpes Simples/diagnóstico , Nariz/virologia , Órbita/virologia , Sinusite/diagnóstico , Tomografia Computadorizada por Raios X/métodos , Idoso de 80 Anos ou mais , DNA Viral/análise , Herpes Simples/virologia , Humanos , Masculino , Nariz/diagnóstico por imagem , Órbita/diagnóstico por imagem , Simplexvirus/genética , Sinusite/virologia
13.
Virol Sin ; 33(6): 484-492, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30570714

RESUMO

We describe the first genome isolation of Middle East respiratory syndrome coronavirus (MERS-CoV) in Kenya. This fatal zoonotic pathogen was first described in the Kingdom of Saudi Arabia in 2012. Epidemiological and molecular evidence revealed zoonotic transmission from camels to humans and between humans. Currently, MERS-CoV is classified by the WHO as having high pandemic potential requiring greater surveillance. Previous studies of MERS-CoV in Kenya mainly focused on site-specific and archived camel and human serum samples for antibodies. We conducted active nationwide cross-sectional surveillance of camels and humans in Kenya, targeting both nasal swabs and plasma samples from 1,163 camels and 486 humans collected from January 2016 to June 2018. A total of 792 camel plasma samples were positive by ELISA. Seroprevalence increased with age, and the highest prevalence was observed in adult camels (82.37%, 95% confidence interval (CI) 79.50-84.91). More female camels were significantly seropositive (74.28%, 95% CI 71.14-77.19) than male camels (P < 0.001) (53.74%, 95% CI 48.48-58.90). Only 11 camel nasal swabs were positive for MERS-CoV by reverse transcription-quantitative PCR. Phylogenetic analysis of whole genome sequences showed that Kenyan MERS-CoV clustered within sub-clade C2, which is associated with the African clade, but did not contain signature deletions of orf4b in African viruses. None of the human plasma screened contained neutralizing antibodies against MERS-CoV. This study confirms the geographically widespread occurrence of MERS-CoV in Kenyan camels. Further one-health surveillance approaches in camels, wildlife, and human populations are needed.


Assuntos
Camelus/virologia , Infecções por Coronavirus/veterinária , Reservatórios de Doenças/veterinária , Genoma Viral , Coronavírus da Síndrome Respiratória do Oriente Médio/genética , Fatores Etários , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Infecções por Coronavirus/transmissão , Estudos Transversais , Reservatórios de Doenças/virologia , Feminino , Humanos , Quênia , Masculino , Coronavírus da Síndrome Respiratória do Oriente Médio/isolamento & purificação , Nariz/virologia , Filogenia , Prevalência , RNA Viral/genética , RNA Viral/isolamento & purificação , Estudos Soroepidemiológicos , Sequenciamento Completo do Genoma , Zoonoses/transmissão , Zoonoses/virologia
15.
PLoS One ; 13(11): e0206679, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30440016

RESUMO

The equine herpesvirus type 1 (EHV-1) ORF1 and ORF71 genes have immune modulatory effects in vitro. Experimental infection of horses using virus mutants with multiple deletions including ORF1 and ORF71 showed promise as vaccine candidates against EHV-1. Here, the combined effects of ORF1 and ORF71 deletions from the neuropathogenic EHV-1 strain Ab4 on clinical disease and host immune response were further explored. Three groups of EHV-1 naïve horses were experimentally infected with the ORF1/71 gene deletion mutant (Ab4ΔORF1/71), the parent Ab4 strain, or remained uninfected. In comparison to Ab4, horses infected with Ab4ΔORF1/71 did not show the initial high fever peak characteristic of EHV-1 infection. Ab4ΔORF1/71 infection had reduced nasal shedding (1/5 vs. 5/5) and, simultaneously, decreased intranasal interferon (IFN)-α, interleukin (IL)-10 and soluble CD14 secretion. However, Ab4 and Ab4ΔORF1/71 infection resulted in comparable viremia, suggesting these genes do not regulate the infection of the mononuclear cells and subsequent viremia. Intranasal and serum anti-EHV-1 antibodies to Ab4ΔORF1/71 developed slightly slower than those to Ab4. However, beyond day 12 post infection (d12pi) serum antibodies in both virus-infected groups were similar and remained increased until the end of the study (d114pi). EHV-1 immunoglobulin (Ig) G isotype responses were dominated by short-lasting IgG1 and long-lasting IgG4/7 antibodies. The IgG4/7 response closely resembled the total EHV-1 specific antibody response. Ex vivo re-stimulation of PBMC with Ab4 resulted in IFN-γ and IL-10 secretion by cells from both infected groups within two weeks pi. Flow cytometric analysis showed that IFN-γ producing EHV-1-specific T-cells were mainly CD8+/IFN-γ+ and detectable from d32pi on. Peripheral blood IFN-γ+ T-cell percentages were similar in both infected groups, albeit at low frequency (~0.1%). In summary, the Ab4ΔORF1/71 gene deletion mutant is less virulent but induced antibody responses and cellular immunity similar to the parent Ab4 strain.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/genética , Herpesvirus Equídeo 1/patogenicidade , Doenças dos Cavalos/imunologia , Doenças dos Cavalos/virologia , Proteínas Virais/genética , Animais , Anticorpos Antivirais/metabolismo , Temperatura Corporal , Citocinas/metabolismo , Feminino , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Cavalos , Imunidade Celular , Imunoglobulina G/metabolismo , Masculino , Mutação , Nariz/imunologia , Nariz/virologia , Distribuição Aleatória , Viremia/imunologia , Viremia/veterinária , Virulência , Eliminação de Partículas Virais
16.
Virol Sin ; 33(5): 410-417, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30311100

RESUMO

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050) samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to detect IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged > 10 years (81.37%) followed by those aged 3.1-10 years (78.65%) and ≤ 3 years (58.19%). Higher prevalence was observed in female (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 2016-2017. Among the sampled population, 840 humans were camel herders. Although we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels.


Assuntos
Anticorpos Antivirais/sangue , Camelus/virologia , Infecções por Coronavirus/veterinária , Monitoramento Epidemiológico/veterinária , Coronavírus da Síndrome Respiratória do Oriente Médio , Animais , Anticorpos Neutralizantes/sangue , Criança , Pré-Escolar , Infecções por Coronavirus/epidemiologia , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Nariz/virologia , Paquistão/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real , Estudos Soroepidemiológicos , Inquéritos e Questionários
17.
Vet Microbiol ; 225: 25-30, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30322528

RESUMO

Bovine viral diarrhea virus (BVDV) belongs to the genus Pestivirus and can cause reproductive problems in cattle. However, there is still a lack of research to clarify its pathogenicity in different gestational periods of sows and its effects in neonates. In this study, 12 gilts divided into groups (G) were experimentally inoculated with the strain BVDV-2 (SV-253) oronasally at a dose of 106·85 TCID50; one group was inoculated 30 days before insemination (G0; n = 2), three groups were inoculated during gestation (first (G1; n = 2), second (G2; n = 3), third (G3; n = 3)), and a fourth was the control group (G4; n = 2). Samples of blood and nasal swabs from the gilts were collected every three days until delivery for a virus neutralization (VN) test, qRT-PCR, and blood count. On the day of delivery, 40% of the neonates were euthanized to obtain tissue and blood samples at necropsy for histopathology and qRT-PCR. The sows were seroconverted between 12 and 33 days after inoculation, and the virus was detected in the blood between 3 and 12 days and on the nasal swab between 6 and 24 days in the G0, G1, G2 and G3 sows but was not detected in piglet tissues, and no significant alterations were found through histopathology. The mean and standard deviation of the mean cycles (Cq) from blood (Cq = 34.87 ± 0.60) and nasal swab (Cq = 34.61 ± 0.87) samples were between 107 and 490 TCID50/ml. Transient infection was demonstrated with a low viral load, but transplacental infection was not possible in gilts.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina Tipo 2/patogenicidade , Transmissão Vertical de Doença Infecciosa/veterinária , Placenta/virologia , Animais , Anticorpos Antivirais/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/imunologia , Bovinos , Vírus da Diarreia Viral Bovina Tipo 2/genética , Vírus da Diarreia Viral Bovina Tipo 2/imunologia , Vírus da Diarreia Viral Bovina Tipo 2/isolamento & purificação , Feminino , Testes de Neutralização , Nariz/virologia , Gravidez , Soroconversão , Suínos , Vacinação
18.
J Vet Diagn Invest ; 30(6): 920-923, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30264670

RESUMO

Influenza A virus (IAV) is a zoonotic pathogen threatening animal and public health; therefore, detection and monitoring of IAV in animal populations are critical components of a surveillance program. Swine are important hosts of IAV, wherein the virus can undergo rapid evolution. Several methods (i.e., nasal swabs, nasal wipes, and oral fluids) have been used to collect samples from swine for IAV surveillance. We utilized nasal wipes made from cotton gauze and multiple, polyester or mixed polyester fabrics to compare performance in the molecular detection and isolation of IAV. In vitro experiments revealed that no polyester or mixed polyester fabric was superior to cotton gauze for molecular IAV detection; however, 3 polyester or mixed polyester fabrics yielded significantly more viable IAV than cotton. In a field trial, both cotton gauze and the polyester or mixed polyester fabric yielded similar proportions of IAV isolates from swine. The results indicate that cotton gauze remains a practical and useful material for swine nasal wipes.


Assuntos
Vírus da Influenza A/isolamento & purificação , Nariz/virologia , Infecções por Orthomyxoviridae/veterinária , Vigilância de Evento Sentinela/veterinária , Manejo de Espécimes/veterinária , Doenças dos Suínos/diagnóstico , Animais , Técnicas e Procedimentos Diagnósticos/instrumentação , Técnicas e Procedimentos Diagnósticos/veterinária , Manejo de Espécimes/instrumentação , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia
19.
J Med Microbiol ; 67(10): 1527-1532, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30136923

RESUMO

PURPOSE: Nontypeable Haemophilus influenzae (NTHi) is a commensal in the human nasopharynx and the cause of pneumonia, meningitis, sinusitis, acute exacerbations of chronic obstructive pulmonary disease and acute otitis media (AOM). AOM is the most common ailment for which antibiotics are prescribed in the United States. With the emergence of new strains of antibiotic-resistant bacteria, finding an effective and broad coverage vaccine to protect against AOM-causing pathogens has become a priority. Mouse models are a cost-effective and efficient way to help determine vaccine efficacy. Here, we describe an NTHi AOM model in C57BL/6J mice, which also utilizes a mouse-adapted H1N1 influenza virus to mimic human coinfection. METHODOLOGY: We tested our coinfection model using a protein vaccine formulation containing protein D, a well-studied NTHi vaccine candidate that can be found in the 10-valent Streptococcus pneumoniae conjugate vaccine. We verified the usefulness of our mouse model by comparing bacterial loads in the nose and ear between protein D-vaccinated and control mice. RESULTS: While there was no measurable difference in nasal bacterial loads, we did detect significant differences in the bacterial loads of ear washes and ear bullae between vaccinated and control mice. CONCLUSION: The results from this study suggest that our NTHi AOM coinfection model is useful for assessing protein vaccines.


Assuntos
Proteínas de Bactérias/imunologia , Proteínas de Transporte/imunologia , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/imunologia , Haemophilus influenzae/imunologia , Imunoglobulina D/imunologia , Lipoproteínas/imunologia , Otite Média/prevenção & controle , Administração Intranasal , Animais , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/genética , Proteínas de Transporte/administração & dosagem , Proteínas de Transporte/genética , Coinfecção/microbiologia , Coinfecção/prevenção & controle , Coinfecção/virologia , Modelos Animais de Doenças , Feminino , Infecções por Haemophilus/microbiologia , Vacinas Anti-Haemophilus/administração & dosagem , Vacinas Anti-Haemophilus/genética , Haemophilus influenzae/genética , Haemophilus influenzae/crescimento & desenvolvimento , Humanos , Imunoglobulina D/administração & dosagem , Imunoglobulina D/genética , Vírus da Influenza A Subtipo H1N1/fisiologia , Lipoproteínas/administração & dosagem , Lipoproteínas/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nariz/microbiologia , Nariz/virologia , Otite Média/imunologia , Otite Média/microbiologia , Vacinas Conjugadas/administração & dosagem , Vacinas Conjugadas/imunologia
20.
Sci Rep ; 8(1): 11411, 2018 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-30061588

RESUMO

The role of nasal and fecal microbiota in viral respiratory infections has not been established. We collected nasal swabs and washes, and fecal samples in a clinical study assessing the effect of probiotic Bifidobacterium animalis subsp. lactis Bl-04 on experimental rhinovirus infection. The nasal and fecal microbiota were characterized by 16S rRNA gene sequencing. The resulting data were compared with nasal inflammatory marker concentrations, viral load, and clinical symptoms. By using unsupervised clustering, the nasal microbiota divided into six clusters. The clusters predominant of Staphylococcus, Corynebacterium/Alloiococcus, Moraxella, and Pseudomonadaceae/Mixed had characteristic inflammatory marker and viral load profiles in nasal washes. The nasal microbiota clusters of subjects before the infection associated with the severity of clinical cold symptoms during rhinovirus infection. Rhinovirus infection and probiotic intervention did not significantly alter the composition of nasal or fecal microbiota. Our results suggest that nasal microbiota may influence the virus load, host innate immune response, and clinical symptoms during rhinovirus infection, however, further studies are needed.


Assuntos
Inflamação/patologia , Microbiota , Nariz/microbiologia , Nariz/virologia , Rhinovirus/fisiologia , Carga Viral , Bactérias/classificação , Biodiversidade , Biomarcadores/metabolismo , Análise por Conglomerados , Fezes/microbiologia , Humanos , Infecções por Picornaviridae/microbiologia , Infecções por Picornaviridae/virologia , Adulto Jovem
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