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Previous research demonstrated the distribution of distinct microbial communities in the equine hindgut surrounding the pelvic flexure. The current study evaluated gene expression in epithelial tissues surrounding the pelvic flexure to characterize patterns that might correlate with microbial distribution. Gene expression was determined by analyzing RNA sequence data from the pelvic flexure, the left and right ventral colon, and the left and right dorsal colon. An average of 18,330 genes were expressed across the five tissues sampled. Most of the genes showed some level of expression in all five tissues. Tissue-restricted patterns of expression were also observed. Genes with restricted expression in the left ventral and left dorsal colons have communication, signaling, and regulatory functions that correlate with their known physiology. In contrast, genes expressed exclusively in the pelvic flexure have diverse functions. The ontology of genes differentially expressed between the pelvic flexure and the surrounding tissues was associated with immune functions and signaling processes. Despite being non-significant, these enrichment trends were reinforced by the functions of statistically significant expression differences between tissues of the hindgut. These results provide insight into the physiology of the equine hindgut epithelium that might influence the microbiota and its distribution.
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BACKGROUND: Equine influenza virus is a common cause of respiratory disease in equids. Few reports describe clinical presentation and disease progression in donkeys. HYPOTHESIS/OBJECTIVES: Describe the clinical and diagnostic findings, outcome, and pathologic lesions associated with influenza pneumonia in donkeys. ANIMALS: Thirteen unvaccinated donkeys ranging from 1 week to 12 years of age and sharing clinical signs and exposure history. METHODS: Retrospective case series. Medical records from June to July 2020 at the Colorado State Veterinary Teaching Hospital and collaborating referring veterinary practices were reviewed. The diagnosis was confirmed by molecular testing, virus isolation, and partial genetic and phylogenetic analysis of the virus. RESULTS: Survival in donkeys <1 year old was 16.6% (1/6) whereas survival in animals >1 year of age was 85.7% (6/7). Hemagglutinin gene sequencing and phylogenetic analysis confirmed a contemporary clade 1 Florida sublineage H3 virus as the causative agent. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinical signs of equine influenza virus infection in donkeys are similar to those observed in horses. Prognosis for survival generally is good, but deaths have been observed especially in foals born to seronegative dams. This finding emphasizes the importance of prenatal vaccination protocols in all equids, including donkeys.
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Enfermedades de los Caballos , Subtipo H3N8 del Virus de la Influenza A , Gripe Humana , Infecciones por Orthomyxoviridae , Caballos , Animales , Humanos , Subtipo H3N8 del Virus de la Influenza A/genética , Equidae , Gripe Humana/epidemiología , Filogenia , Estudios Retrospectivos , Hospitales Veterinarios , Enfermedades de los Caballos/epidemiología , Hospitales de Enseñanza , Infecciones por Orthomyxoviridae/epidemiología , Infecciones por Orthomyxoviridae/veterinaria , Brotes de Enfermedades/veterinaria , Progresión de la EnfermedadRESUMEN
Detection of Leptospira interrogans is difficult as a result of intermittent leptospiruria and brief leptospiremia. Hence, diagnosis relies heavily on serologic testing, the reference method of which is the microscopic agglutination test (MAT). In horses, clinical leptospirosis has been associated with abortion, recurrent uveitis, and sporadic cases of hepatic and renal disease. Little information exists on the seroprevalence of antibodies to L. interrogans in equids in the United States; past nationwide studies suggest that the seroprevalence in some areas is as high as 77% (reciprocal titer ≥ 100). We tested sera from 124 apparently healthy horses previously submitted for equine infectious anemia (EIA) serology using MAT for 6 serovars-Bratislava, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, and Pomona. When using a reciprocal MAT titer cutoff of ≥ 100, 102 of 124 (82%) of the samples were positive for at least one serovar. Seropositivity was significantly associated with increasing age. Query of specimens from clinical cases submitted to the Colorado State University Veterinary Diagnostic Laboratory for MAT since 2010 indicated significantly greater seroprevalence (p = 0.015) of pathogenic serovar Pomona in clinical cases compared to sera submitted from healthy equids for routine EIA testing. Information from our diagnostic laboratory submission forms also suggests a correlation between uveitis or other ophthalmic problems and serovar Pomona.
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Enfermedades de los Caballos/epidemiología , Leptospirosis/veterinaria , Factores de Edad , Animales , Anticuerpos Antibacterianos/sangre , Colorado/epidemiología , Femenino , Enfermedades de los Caballos/microbiología , Caballos , Leptospira , Leptospirosis/epidemiología , Leptospirosis/microbiología , Masculino , Prevalencia , Estudios Seroepidemiológicos , SerogrupoRESUMEN
Detection of equine acute kidney injury (AKI) is hindered by limited markers of early renal damage in horses. N-acetyl-ß-D-glucosaminidase (NAG), a lysosomal enzyme in renal tubular epithelium released into urine during tubular insult, has shown promise for early identification of AKI in humans and other species. We validated an assay for NAG in equine urine and measured urinary NAG in 7 azotemic and 7 non-azotemic client-owned adult horses. The enzymatic NAG assay was validated using within- and between-run coefficients of variation (CVs), recovery following standard addition, and linearity of dilution. Intra- and inter-run CVs (21% and 3.2%, respectively), average recovery following standard addition (99-109%), and linearity under serial dilution (R2 = 0.997) were satisfactory. Urine NAG index was significantly correlated with urinary fractional excretion of sodium (FENa; ρ = 0.76, p < 0.001) and plasma creatinine (ρ = 0.74, p = 0.001). Median urine NAG indices were higher in azotemic horses (p = 0.006), in horses with increased urinary FENa (p = 0.006), and in horses with increased urine gamma-glutamyl transferase index (p = 0.032). Urine NAG can be measured in horses and shows positive correlation with 2 current renal biomarkers. Additional work is needed to establish normal equine reference intervals and characterize the increase of urine NAG index in horses in relation to tubular injury.
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Acetilglucosaminidasa/orina , Lesión Renal Aguda/veterinaria , Enfermedades de los Caballos/diagnóstico , Lesión Renal Aguda/diagnóstico , Lesión Renal Aguda/orina , Animales , Biomarcadores/orina , Creatinina/sangre , Femenino , Enfermedades de los Caballos/orina , Caballos , Humanos , Masculino , Sodio/orina , gamma-Glutamiltransferasa/orinaRESUMEN
For decades the horse has been viewed as an isolated or "dead-end" host for influenza A viruses, with equine influenza virus being considered as relatively stable genetically. Although equine influenza viruses are genetically more stable than those of human lineage, they are by no means in evolutionary stasis. Moreover, recent transmission of equine-lineage influenza viruses to dogs also challenges the horse's status as a dead-end host. This article reviews recent developments in the epidemiology and evolution of equine influenza virus. In addition, the clinical presentation of equine influenza infection, diagnostic techniques, and vaccine recommendations are briefly summarized.
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Enfermedades de los Caballos/prevención & control , Enfermedades de los Caballos/virología , Virus de la Influenza A/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Animales , Enfermedades de los Perros/virología , Perros , Caballos , Virus de la Influenza A/genética , Virus de la Influenza A/inmunología , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/inmunología , Infecciones por Orthomyxoviridae/prevención & control , Infecciones por Orthomyxoviridae/virologíaRESUMEN
With the widespread use of a recently developed canine influenza virus (CIV) H3N8 vaccine, continual molecular evaluation of circulating CIVs is necessary for monitoring antigenic drift. The aim of this project was to further describe the genetic evolution of CIV, as well as determine any genetic variation within potential antigenic regions that might result in antigenic drift. To this end, the hemagglutinin gene of 19 CIV isolates from dogs residing in Colorado, New York, and South Carolina humane shelters was sequenced and compared to CIV strains isolated during 2003-2012. Phylogenetic analysis suggests that CIV might be diverging into two geographically distinct lineages. Using a mixed-effects model for evolution and single likelihood ancestor counting methods, several amino acid sites were found to be undergoing selection pressure. Additionally, a total of six amino acid changes were observed in two possible antigenic sites for CIVs isolated from Colorado and New York humane shelters between 2009 and 2011. As CIV isolates might be diverging into geographically distinct lineages, further experiments are warranted to determine the extent of antigenic drift occurring within circulating CIV.
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Enfermedades de los Perros/virología , Evolución Molecular , Variación Genética , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Subtipo H3N8 del Virus de la Influenza A/enzimología , Subtipo H3N8 del Virus de la Influenza A/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Sustitución de Aminoácidos , Animales , Análisis por Conglomerados , Colorado , Perros , Epítopos/genética , Subtipo H3N8 del Virus de la Influenza A/genética , Datos de Secuencia Molecular , New York , Infecciones por Orthomyxoviridae/virología , Filogeografía , ARN Viral/genética , Análisis de Secuencia de ADN , Homología de Secuencia , South CarolinaRESUMEN
Phylogenetic analyses indicate that canine influenza viruses (CIVs) (H3N8) evolved from contemporary equine influenza virus (EIV). Despite the genetic relatedness of EIV and CIV, recent evidence suggests that CIV is unable to infect, replicate, and spread among susceptible horses. To determine whether equine H3N8 viruses have equally lost the ability to infect, cause disease, and spread among dogs, we evaluated the infectivity and transmissibility of a recent Florida sublineage EIV isolate in dogs. Clinical signs, nasal virus shedding, and serological responses were monitored in dogs for 21 days after inoculation. Real-time reverse transcription-PCR and hemagglutination inhibition assays showed that both the viruses have maintained the ability to infect and replicate in dogs and result in seroconversion. Transmission of EIV from infected to sentinel dogs, however, was restricted. Furthermore, both CIV and EIV exhibited similar sialic acid- α 2,3-gal receptor-binding preferences upon solid-phase binding assays. The results of the in vivo experiments reported here suggesting that dogs are susceptible to EIV and previous reports by members of our laboratory showing limited CIV infection in horses have been mirrored in CIV and EIV infections studies in primary canine and equine respiratory epithelial cells.
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Sustained transmission of canine Influenza A virus (CIV) H3N8 among U.S. dogs underscores the threat influenza continues to pose to canine health. Because rapid and accurate detection of infection is critical to the diagnosis and control of CIV, the 2 main objectives of the current study were to estimate and compare the sensitivities of CIV testing methods on canine swab samples and to evaluate the performance of Flu Detect™ (Synbiotics Corp., Kansas City, MO) for detecting CIV nasal shedding in high-risk shelter dogs. To address the first objective, nasal and pharyngeal swab samples were collected from 124 shelter and household dogs seen by Colorado State University Veterinary Teaching Hospital clinicians for canine infectious respiratory disease between April 2006 and March 2007 and tested for CIV shedding using virus isolation, the rapid influenza diagnostic test Directigen Flu A+B™ (BD Diagnostic Systems, Sparks, MD), and real-time reverse transcription polymerase chain reaction (RT-PCR). For the second objective, 1,372 dogs with unknown respiratory health status were sampled from 6 U.S. shelters from December 2009 to November 2010. Samples were tested for presence of CIV using real-time RT-PCR and Flu Detect. Using a stochastic latent class modeling approach, the median sensitivities of virus isolation, rapid influenza diagnostic test, and real-time RT-PCR were 72%, 65%, and 95%, respectively. The Flu Detect test performed poorly for detecting CIV nasal shedding compared to real-time RT-PCR. In conclusion, the real-time RT-PCR has the highest sensitivity for detecting virus nasal shedding and can be used as a rapid diagnostic test for CIV.
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Enfermedades de los Perros/diagnóstico , Subtipo H3N8 del Virus de la Influenza A/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Animales , Enfermedades de los Perros/virología , Perros , Infecciones por Orthomyxoviridae/diagnóstico , Infecciones por Orthomyxoviridae/virología , Sensibilidad y EspecificidadRESUMEN
We conducted a cross-sectional convenience sampling study of dogs racing in the 2010 Iditarod to determine the seroprevalence of canine influenza virus (CIV) in the sled dog population. Questionnaires were completed detailing medical and CIV vaccination history, kennel size and location, travel history, and social interactions for each team. A total of 399 dogs were tested for CIV antibodies by hemagglutination inhibition assay. None of these, including 39 samples from dogs reported as CIV vaccinated, were seropositive for CIV antibodies. All of the vaccinated dogs were also negative on virus microneutralization assay. Risk factors for CIV seropositivity could not be determined due to a lack of positive samples. This is the first published study investigating the prevalence of CIV in sled dogs and additional studies are warranted to assess CIV infection among racing sled dogs and to evaluate the ecology of CIV and the vaccine efficacy in this population of dogs.
Séroprévalence du virus de la grippe canine (H3N8) chez les chiens de traîneau de la course Iditarod. Nous avons réalisé une étude par sondage des chiens de la course Iditarod 2010 afin de déterminer la séroprévalence du virus de la grippe canine (VGC) dans la population de chiens de traîneau. Les questionnaires remplis fournissaient des détails complets sur les antécédents médicaux et l'historique de vaccination contre le VGC, la taille du chenil et l'emplacement, l'historique des déplacements et les interactions sociales entre chaque équipe. Un total de 399 chiens a été testé pour les anticorps du VGC par un test d'inhibition de l'hémagglutination. Aucun de ces tests, incluant les 39 échantillons provenant de chiens déclarés comme étant vaccinés contre le VGC, étaient séropositifs pour les anticorps de la VGC. Tous les chiens vaccinés ont aussi eu des résultats négatifs au test de microneutralisation. Les facteurs de risque pour la séropositivité au VGC n'ont pas pu être déterminés en raison d'une absence d'échantillons positifs. Il s'agit de la première étude publiée étudiant la prévalence du VGC chez les chiens de traîneaux et des études additionnelles sont nécessaires pour évaluer l'infection par le VGC chez les chiens de traîneau de course et déterminer l'écologie du VGC et l'efficacité du vaccin chez cette population de chiens.(Traduit par Isabelle Vallières).
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Anticuerpos Antivirales/sangre , Subtipo H3N8 del Virus de la Influenza A/inmunología , Infecciones por Orthomyxoviridae/veterinaria , Animales , Canadá/epidemiología , Estudios Transversales , Perros , Femenino , Pruebas de Inhibición de Hemaglutinación/veterinaria , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/inmunología , Masculino , Infecciones por Orthomyxoviridae/epidemiología , Estudios SeroepidemiológicosRESUMEN
OBJECTIVE: To evaluate whether an equine-derived canine H3N8 influenza A virus was capable of infecting and transmitting disease to ponies. ANIMALS: 20 influenza virus-seronegative 12- to 24-month-old ponies. PROCEDURES: 5 ponies were inoculated via aerosol exposure with 10(7) TCID(50) of A/Canine/Wyoming/86033/07 virus (Ca/WY)/pony. A second group of 5 ponies (positive control group) was inoculated via aerosol exposure with a contemporary A/Eq/Colorado/10/07 virus (Eq/CO), and 4 sham-inoculated ponies served as a negative control group. To evaluate the potential for virus transmission, ponies (3/inoculation group) were introduced 2 days after aerosol exposure and housed with Ca/WY- and Eq/CO-inoculated ponies to serve as sentinel animals. Clinical signs, nasal virus shedding, and serologic responses to inoculation were monitored in all ponies for up to 21 days after viral inoculation. Growth and infection characteristics of viruses were examined by use of Madin-Darby canine kidney cells and primary equine and canine respiratory epithelial cells. RESULTS: Ponies inoculated with Ca/WY had mild changes in clinical appearance, compared with results for Eq/CO-inoculated ponies. Additionally, Ca/WY inoculation induced significantly lower numbers for copies of the matrix gene in nasal secretions and lower systemic antibody responses in ponies than did Eq/CO inoculation. The Ca/WY isolate was not transmitted to sentinel ponies. CONCLUSIONS AND CLINICAL RELEVANCE: Inoculation of ponies with the canine H3N8 isolate resulted in mild clinical disease, minimal nasal virus shedding, and weak systemic antibody responses, compared with responses after inoculation with the equine H3N8 influenza isolate. These results suggested that Ca/WY has not maintained infectivity for ponies.
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Anticuerpos Antivirales/sangre , Células Epiteliales/virología , Enfermedades de los Caballos/virología , Subtipo H3N8 del Virus de la Influenza A/patogenicidad , Infecciones por Orthomyxoviridae/veterinaria , Tráquea/virología , Animales , Formación de Anticuerpos , Células Cultivadas , Enfermedades de los Perros/patología , Enfermedades de los Perros/transmisión , Enfermedades de los Perros/virología , Perros , Enfermedades de los Caballos/patología , Enfermedades de los Caballos/transmisión , Caballos , Subtipo H3N8 del Virus de la Influenza A/aislamiento & purificación , Datos de Secuencia Molecular , Infecciones por Orthomyxoviridae/patología , Infecciones por Orthomyxoviridae/transmisión , Infecciones por Orthomyxoviridae/virología , Distribución Aleatoria , Tráquea/citología , Esparcimiento de VirusRESUMEN
Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, ß-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, ß-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-ß, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions.
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Caballos/inmunología , Mucosa Respiratoria/inmunología , 2',5'-Oligoadenilato Sintetasa/genética , Animales , Secuencia de Bases , Diferenciación Celular , Células Cultivadas , Quimiocinas/genética , Citocinas/genética , Cartilla de ADN/genética , Femenino , Proteínas de Unión al GTP/genética , Expresión Génica , Caballos/genética , Caballos/metabolismo , Inmunidad Innata , Lactoferrina/genética , Masculino , Proteínas de Resistencia a Mixovirus , ARN Mensajero/genética , ARN Mensajero/metabolismo , Mucosa Respiratoria/citología , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo , beta-Defensinas/genéticaRESUMEN
Infection of dogs with canine influenza virus (CIV) is considered widespread throughout the United States following the first isolation of CIV in 2004. While vaccination against influenza A infection is a common and important practice for disease control, antiviral therapy can serve as a valuable adjunct in controlling the impact of the disease. In this study, we examined the antiviral activity of nitazoxanide (NTZ) and tizoxanide (TIZ) against three CIV isolates in vitro. NTZ and TIZ inhibited virus replication of all CIVs with 50% and 90% inhibitory concentrations ranging from 0.17 to 0.21 µM and from 0.60 to 0.76 µM, respectively. These results suggest that NTZ and TIZ are effective against CIV and may be useful for treatment of canine influenza in dogs but further investigation of the in vivo efficacy against CIV as well as the drug's potential for toxicity in dogs is needed.
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Equine influenza virus remains an important problem in horses despite extensive use of vaccination. Efficacy of equine influenza vaccination depends on the onset and duration of protective immunity, and appropriate strain specificity of the immune response. This study was designed to test the protective immunity resulting from vaccination with the North American commercial ALVAC equine influenza vaccine (RECOMBITEK Influenza, Merial, USA)(1) against challenge with American lineage influenza viruses. In experiment 1, 12 ponies were vaccinated twice, at a 35 day interval, using the ALVAC-influenza vaccine expressing the HA genes of influenza A/eq/Newmarket/2/93 and A/eq/Kentucky/94 (H3N8), and 11 ponies served as unvaccinated controls. Six months after the second vaccination, all ponies were challenged with A/eq/Kentucky/91. In experiment 2, 10 ponies received one dose of the ALVAC-influenza vaccine, 10 ponies served as unvaccinated controls, and all ponies were challenge infected with A/equine/Ohio/03, 14 days after vaccination. Parameters studied included serological responses, and clinical disease and nasal viral shedding following challenge infection. In experiment 1, following the two-dose regimen, vaccinated ponies generated high titered anti-influenza virus IgGa and IgGb antibody responses to vaccination and demonstrated statistically significant clinical and virological protection to challenge infection compared to controls. Infection with A/eq/Kentucky/91 produced unusually severe signs in ponies in the control group, requiring therapy with NSAID's and antibiotics, and leading to the euthanasia of one pony. In experiment 2 following the one-dose regimen, vaccinates generated IgGa responses pre-challenge, and anamnestic IgGa and IgGb responses after challenge. Vaccinates demonstrated statistically significant clinical and virological protection to challenge infection compared to controls. The results of this study clearly demonstrate the early onset, and 6-month duration of protective immunity resulting from ALVAC-influenza vaccination against challenge with American lineage equine influenza viruses.
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Enfermedades de los Caballos/prevención & control , Subtipo H3N8 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/uso terapéutico , Infecciones por Orthomyxoviridae/veterinaria , Animales , Anticuerpos Antivirales/sangre , Formación de Anticuerpos/inmunología , Virus de la Viruela de los Canarios , Enfermedades de los Caballos/inmunología , Enfermedades de los Caballos/virología , Caballos/inmunología , Vacunas contra la Influenza/inmunología , Masculino , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/prevención & control , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/uso terapéutico , Esparcimiento de Virus/inmunologíaRESUMEN
In the late 1990s, triple reassortant H3N2 influenza A viruses emerged and spread widely within the swine population of the United States. We have shown previously that an isolate representative of this lineage of viruses, A/Swine/Minnesota/593/99 (Sw/MN), has higher infectivity and accelerated replication kinetics in pigs, compared to a human-lineage H3N2 virus isolated from a pig during the same time period, A/Swine/Ontario/00130/97 (Sw/ONT [Landolt, G.A., Karasin, A.I., Phillips, L., Olsen, C.W., 2003. Comparison of the pathogenesis of two genetically different H3N2 influenza A viruses in pigs. J. Clin. Microbiol. 41, 1936-1941]). Additional in vivo experiments using reverse genetics-generated reassortant viruses demonstrated that these phenotypes are dependent upon the HA and/or NA genes (Landolt, G.A., Karasin, A.I., Schutten, M.M., Olsen, C.W., 2006. Restricted infectivity of a human-lineage H3N2 influenza A virus in pigs is hemagglutinin and neuraminidase gene dependent. J. Clin. Microbiol. 44, 297-301). To further study the infectivity of influenza viruses for pigs, we developed a primary swine respiratory epithelial cell (SREC) culture model. In SRECs, Sw/MN infects a significantly higher number of cells compared to Sw/ONT. Using reverse genetics-generated Sw/MN x Sw/ONT reassortant viruses we demonstrate that the infectivity phenotypes of these viruses in SRECs are strongly dependent upon the HA gene. Using chimeras and point directed mutations within the HA genes, we have identified amino acids that, either alone or in combination with other amino acids, impact infectivity. In particular, amino acid 138 is the dominant factor in determining infectivity levels in SRECs.
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Aminoácidos/genética , Células Epiteliales/virología , Glicoproteínas Hemaglutininas del Virus de la Influenza/química , Subtipo H3N2 del Virus de la Influenza A/patogenicidad , Virus de la Influenza A/patogenicidad , Sistema Respiratorio/citología , Aminoácidos/química , Animales , Línea Celular , Células Cultivadas , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Humanos , Subtipo H3N2 del Virus de la Influenza A/genética , Virus de la Influenza A/clasificación , Virus de la Influenza A/genética , Infecciones por Orthomyxoviridae/veterinaria , Infecciones por Orthomyxoviridae/virología , Mutación Puntual , PorcinosRESUMEN
Influenza is a highly contagious disease that has burdened both humans and animals since ancient times. In humans, the most dramatic consequences of influenza are associated with periodically occurring pandemics. Pandemics require the emergence of an antigenically novel virus to which the majority of the population lacks protective immunity. Historically, influenza A viruses from animals have contributed to the generation of human pandemic viruses and they may do so again in the future. It is, therefore, critical to understand the epidemiological and molecular mechanisms that allow influenza A viruses to cross species barriers. This review summarizes the current knowledge of influenza ecology, and the viral factors that are thought to determine influenza A virus species specificity.
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Virus de la Influenza A/patogenicidad , Gripe Humana/virología , Zoonosis/virología , Animales , Humanos , Especificidad de la EspecieRESUMEN
Acute poisoning and envenomation often represent a diagnostic and therapeutic challenge. Although identification ultimately may benefit the affected animal, treatment frequently must commence before an etiologic diagnosis is established. Therefore, the goals for the management of acutely intoxicated horses must be focused on emergency intervention and stabilization of the patient, prevention of further exposure, and aggressive decontamination. This article reviews the treatment steps that should be considered during the management of horses experiencing poisoning or envenomation.
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Enfermedades de los Caballos/terapia , Intoxicación/veterinaria , Animales , Manejo de la Enfermedad , Enfermedades de los Caballos/diagnóstico , Caballos , Intoxicación/diagnóstico , Intoxicación/terapia , Mordeduras de Serpientes/diagnóstico , Mordeduras de Serpientes/terapia , Mordeduras de Serpientes/veterinariaRESUMEN
Influenza A viruses cause pandemics at sporadic intervals. Pandemic viruses can potentially be introduced into the human population through in toto transfer of an avian influenza virus or through reassortment between avian and human strains. Pigs are believed to play a central role in the creation of pandemic viruses through reassortment because of their susceptibility to infection with both avian and human influenza viruses. However, we recently found that a human-lineage H3N2 influenza virus was highly restricted in its ability to infect pigs after intranasal inoculation. We hypothesized that this restricted infectivity phenotype was controlled by the hemagglutinin (HA) and neuraminidase (NA). To test this, we infected pigs with reverse genetics-created HA plus NA reassortant viruses. Specifically, introduction of the HA and NA genes of a contemporary H3N2 swine virus into the genetic background of the wholly human virus resulted in a significant increase in virus shedding and pathogenicity. These data indicate that the HA/NA can play important roles in controlling human influenza virus infectivity in pigs. The results further support the premise that a barrier exists to human influenza virus infection in pigs, which may limit the role of pigs in pandemic virus creation through reassortment of human and avian influenza viruses.
Asunto(s)
Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Subtipo H3N2 del Virus de la Influenza A/patogenicidad , Gripe Humana/virología , Neuraminidasa/genética , Infecciones por Orthomyxoviridae/veterinaria , Enfermedades de los Porcinos/fisiopatología , Animales , Línea Celular , Pruebas de Inhibición de Hemaglutinación , Glicoproteínas Hemaglutininas del Virus de la Influenza/metabolismo , Humanos , Neuraminidasa/metabolismo , Infecciones por Orthomyxoviridae/fisiopatología , Infecciones por Orthomyxoviridae/virología , Enfermedades de los Porcinos/virología , Replicación Viral , Esparcimiento de VirusRESUMEN
OBJECTIVE: To evaluate sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay performed on pooled nasal swab specimens, compared with virus isolation performed on individual nasal swab specimens by use of 2 cell culture lines for detection of swine influenza A viruses. SAMPLE POPULATION: 900 nasal swab specimens obtained from pigs at an abattoir and 62 nasal swab specimens submitted for diagnostic testing. PROCEDURES: Primers were chosen to amplify a conserved portion of the influenza virus matrix gene. Assay sensitivity was initially determined by testing serial dilutions of various subtypes of swine influenza viruses. Sensitivity and specificity were confirmed by use of nasal swab specimens with or without addition of known concentrations of influenza virus and further validated by testing nasal swab specimens obtained through an abattoir surveillance program or submitted for diagnostic testing. Aliquots of specimens were pooled in sets of 10, and results of real-time RT-PCR assays were compared with results of virus isolation of individual specimens in Madin Darby canine kidney (MDCK) and mink lung (Mv1Lu) cells. RESULTS: Real-time RT-PCR assay was highly specific (100%) and sensitive (88% to 100%). Among the 16 viruses isolated, 3 grew only in Mv1Lu cells and 3 grew only in MDCK cells. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that real-time RT-PCR assay is a fast and accurate test for screening numerous nasal swab specimens for swine influenza virus. Some viruses were isolated in only MDCK or Mv1Lu cells, indicating that use of >1 cell line may be required to isolate a broad range of influenza A viruses.
Asunto(s)
Técnicas de Cultivo de Célula/veterinaria , Virus de la Influenza A/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Enfermedades de los Porcinos/diagnóstico , Animales , Línea Celular , ADN Viral/aislamiento & purificación , Perros , Visón , Mucosa Nasal/virología , Infecciones por Orthomyxoviridae/diagnóstico , Sensibilidad y Especificidad , PorcinosRESUMEN
In 1997 and 1998, H3N2 influenza A viruses emerged among pigs in North America. Genetic analyses of the H3N2 isolates demonstrated that they had distinctly different genotypes. The most commonly isolated viruses in the United States have a triple-reassortant genotype, with the hemagglutinin, neuraminidase, and PB1 polymerase genes being of human influenza virus origin, the nucleoprotein, matrix, and nonstructural genes being of classical swine influenza virus origin, and the PA and PB2 polymerase genes being of avian influenza virus origin. In contrast, a wholly human H3N2 virus was isolated from a single baby pig in Ontario, Canada, in 1997, but it did not spread within the swine population. Genetic differences between this wholly human virus and the triple-reassortant viruses may affect their replication efficiencies in pigs. In the present study we compared the pathogenicities and replication kinetics of the wholly human virus and a triple-reassortant virus in 7-week-old pigs that were infected intranasally with 2 x 10(3) to 2 x 10(6) 50% tissue culture infective doses of virus. Our results demonstrate that the wholly human virus replicated to significantly lower titers and that the onset of virus shedding was delayed compared to the replication titers and the time of onset of virus shedding in triple-reassortant viruses. In addition, infection with the triple-reassortant virus was associated with moderate to severe gross pathological and histological pulmonary lesions, while infection with the wholly human virus induced only mild pulmonary changes.