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1.
Vet Res ; 52(1): 103, 2021 Jul 08.
Artículo en Inglés | MEDLINE | ID: mdl-34238364

RESUMEN

Nocardioform placentitis (NP) continues to result in episodic outbreaks of abortion and preterm birth in mares and remains a poorly understood disease. The objective of this study was to characterize the transcriptome of the chorioallantois (CA) of mares with NP. The CA were collected from mares with confirmed NP based upon histopathology, microbiological culture and PCR for Amycolatopsis spp. Samples were collected from the margin of the NP lesion (NPL, n = 4) and grossly normal region (NPN, n = 4). Additionally, CA samples were collected from normal postpartum mares (Control; CRL, n = 4). Transcriptome analysis identified 2892 differentially expressed genes (DEGs) in NPL vs. CRL and 2450 DEGs in NPL vs. NPN. Functional genomics analysis elucidated that inflammatory signaling, toll-like receptor signaling, inflammasome activation, chemotaxis, and apoptosis pathways are involved in NP. The increased leukocytic infiltration in NPL was associated with the upregulation of matrix metalloproteinase (MMP1, MMP3, and MMP8) and apoptosis-related genes, such as caspases (CASP3 and CASP7), which could explain placental separation associated with NP. Also, NP was associated with downregulation of several placenta-regulatory genes (ABCG2, GCM1, EPAS1, and NR3C1), angiogenesis-related genes (VEGFA, FLT1, KDR, and ANGPT2), and glucose transporter coding genes (GLUT1, GLUT10, and GLUT12), as well as upregulation of hypoxia-related genes (HIF1A and EGLN3), which could elucidate placental insufficiency accompanying NP. In conclusion, our findings revealed for the first time, the key regulators and mechanisms underlying placental inflammation, separation, and insufficiency during NP, which might lead to the development of efficacious therapies or diagnostic aids by targeting the key molecular pathways.


Asunto(s)
Corioamnionitis/veterinaria , Infecciones por Bacterias Grampositivas/veterinaria , Enfermedades de los Caballos/inmunología , Transcriptoma , Actinobacteria/aislamiento & purificación , Amycolatopsis/aislamiento & purificación , Animales , Corioamnionitis/inmunología , Corioamnionitis/microbiología , Femenino , Perfilación de la Expresión Génica/veterinaria , Infecciones por Bacterias Grampositivas/inmunología , Infecciones por Bacterias Grampositivas/microbiología , Enfermedades de los Caballos/microbiología , Caballos , Embarazo
2.
J Virol ; 93(23)2019 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-31511388

RESUMEN

Equid herpesvirus 1 (EHV-1) is a viral pathogen of horse populations worldwide spread by the respiratory route and is known for causing outbreaks of neurologic syndromes and abortion storms. Previously, we demonstrated that an EHV-1 strain of the neuropathogenic genotype, T953, downregulates the beta interferon (IFN-ß) response in vitro in equine endothelial cells (EECs) at 12 h postinfection (hpi). In the present study, we explored the molecular correlates of this inhibition as clues toward an understanding of the mechanism. Data from our study revealed that EHV-1 infection of EECs significantly reduced both Toll-like receptor 3 (TLR3) and TLR4 mRNA expression at 6 hpi and 12 hpi. While EHV-1 was able to significantly reduce IRF9 mRNA at both 6 hpi and 12 hpi, the virus significantly reduced IFN regulatory factor 7 (IRF7) mRNA only at 12 hpi. EHV-1 did not alter the cellular level of Janus-activated kinase 1 (JAK1) at any time point. However, EHV-1 reduced the cellular level of expression of tyrosine kinase 2 (TYK2) at 12 hpi. Downstream of JAK1-TYK2 signaling, EHV-1 blocked the phosphorylation and activation of signal transducer and activator of transcription 2 (STAT2) when coincubated with exogenous IFN, at 12 hpi, although not at 3 or 6 hpi. Immunofluorescence staining revealed that the virus prevented the nuclear translocation of STAT2 molecules, confirming the virus-mediated inhibition of STAT2 activation. The pattern of suppression of phosphorylation of STAT2 by EHV-1 implicated viral late gene expression. These data help illuminate how EHV-1 strategically inhibits the host innate immune defense by limiting steps required for type I IFN sensitization and induction.IMPORTANCE To date, no commercial vaccine label has a claim to be fully protective against the diseases caused by equid herpesvirus 1 (EHV-1), especially the neurologic form. The interferon (IFN) system, of which type I IFN is of great importance, still remains a viable immunotherapeutic option against EHV-1 infection. The type I IFN system has been exploited successfully to treat other viral infections, such as chronic hepatitis B and C in humans. The current state of research on how EHV-1 interferes with the protective effect of type I IFN has indicated transient induction of type I IFN production followed by a rapid shutdown in vitro in equine endothelial cells (EECs). The significance of our study is the identification of certain steps in the type I IFN signaling pathway targeted for inhibition by EHV-1. Understanding this pathogen-host relationship is essential for the long-term goal of developing effective immunotherapy against EHV-1.


Asunto(s)
Células Endoteliales/metabolismo , Células Endoteliales/virología , Infecciones por Herpesviridae/inmunología , Infecciones por Herpesviridae/metabolismo , Herpesvirus Équido 1/inmunología , Interferón Tipo I/metabolismo , Animales , Regulación de la Expresión Génica , Hepatitis B Crónica , Infecciones por Herpesviridae/virología , Herpesvirus Équido 1/genética , Enfermedades de los Caballos/virología , Caballos , Interacciones Huésped-Patógeno , Humanos , Inmunidad Innata , Janus Quinasa 1/metabolismo , ARN Mensajero/metabolismo , Factor de Transcripción STAT2/metabolismo , Transducción de Señal , TYK2 Quinasa/metabolismo , Receptor Toll-Like 3/metabolismo , Receptor Toll-Like 4/metabolismo
3.
PLoS Pathog ; 11(1): e1004610, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25569288

RESUMEN

Lentiviral Envelope (Env) antigenic variation and related immune evasion present major hurdles to effective vaccine development. Centralized Env immunogens that minimize the genetic distance between vaccine proteins and circulating viral isolates are an area of increasing study in HIV vaccinology. To date, the efficacy of centralized immunogens has not been evaluated in the context of an animal model that could provide both immunogenicity and protective efficacy data. We previously reported on a live-attenuated (attenuated) equine infectious anemia (EIAV) virus vaccine, which provides 100% protection from disease after virulent, homologous, virus challenge. Further, protective efficacy demonstrated a significant, inverse, linear relationship between EIAV Env divergence and protection from disease when vaccinates were challenged with viral strains of increasing Env divergence from the vaccine strain Env. Here, we sought to comprehensively examine the protective efficacy of centralized immunogens in our attenuated vaccine platform. We developed, constructed, and extensively tested a consensus Env, which in a virulent proviral backbone generated a fully replication-competent pathogenic virus, and compared this consensus Env to an ancestral Env in our attenuated proviral backbone. A polyvalent attenuated vaccine was established for comparison to the centralized vaccines. Additionally, an engineered quasispecies challenge model was created for rigorous assessment of protective efficacy. Twenty-four EIAV-naïve animals were vaccinated and challenged along with six-control animals six months post-second inoculation. Pre-challenge data indicated the consensus Env was more broadly immunogenic than the Env of the other attenuated vaccines. However, challenge data demonstrated a significant increase in protective efficacy of the polyvalent vaccine. These findings reveal, for the first time, a consensus Env immunogen that generated a fully-functional, replication-competent lentivirus, which when experimentally evaluated, demonstrated broader immunogenicity that does not equate to higher protective efficacy.


Asunto(s)
Anemia Infecciosa Equina/prevención & control , Caballos/inmunología , Virus de la Anemia Infecciosa Equina/inmunología , Vacunas Atenuadas/uso terapéutico , Proteínas del Envoltorio Viral/inmunología , Secuencia de Aminoácidos , Animales , Variación Antigénica/inmunología , Secuencia de Bases , Variación Genética , Virus de la Anemia Infecciosa Equina/genética , Datos de Secuencia Molecular , Filogenia , Resultado del Tratamiento , Proteínas del Envoltorio Viral/genética , Vacunas Virales/uso terapéutico
4.
Vet Clin North Am Equine Pract ; 30(3): 641-58, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25300636

RESUMEN

Lawsonia intracellularis is the etiologic agent for equine proliferative enteropathy (EPE), which typically affects weanling and yearling horses. In North America, EPE cases often occur between August and January, although cases outside of this time frame have been reported. Clinical signs of EPE are usually nonspecific and include lethargy, pyrexia, anorexia, peripheral edema, weight loss, colic, and diarrhea. Diagnosis is based on the presence of hypoproteinemia and hypoalbuminemia along with clinical signs and positive commercial serologic and/or molecular testing. Treatment requires the use of antimicrobials with good intracellular penetration and supportive care to prevent or decrease secondary complications.


Asunto(s)
Infecciones por Desulfovibrionaceae/microbiología , Infecciones por Desulfovibrionaceae/veterinaria , Enfermedades de los Caballos/microbiología , Enfermedades Intestinales/veterinaria , Lawsonia (Bacteria)/aislamiento & purificación , Animales , Enfermedades de los Caballos/tratamiento farmacológico , Caballos , Enfermedades Intestinales/tratamiento farmacológico , Enfermedades Intestinales/microbiología
5.
Vet Clin North Am Equine Pract ; 30(3): 561-77, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25441114

RESUMEN

In the absence of an effective vaccine, the success of the test and removal approach for the control of equine infectious anemia (EIA) cannot be overstated, at least in those areas where testing has been traditionally routine. This article addresses 4 main aspects: what has been learned about EIA virus, host control of its replication, and inapparent carriers; international status regarding the control of EIA; diagnostic and laboratory investigation; and reducing the spread of blood-borne infections by veterinarians. An attempt is made to put these issues into practical contemporary perspectives for the equine practitioner.


Asunto(s)
Anemia Infecciosa Equina/prevención & control , Enfermedades de los Caballos/prevención & control , Virus de la Anemia Infecciosa Equina/aislamiento & purificación , Animales , Erradicación de la Enfermedad , Equidae , Enfermedades de los Caballos/virología , Caballos
6.
PLoS One ; 19(6): e0306211, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38905290

RESUMEN

[This corrects the article DOI: 10.1371/journal.pone.0290778.].

7.
Equine Vet J ; 55(5): 905-915, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-36397207

RESUMEN

BACKGROUND: Intra-articular (IA) corticosteroids are regularly used in equine athletes for the control of joint inflammation. OBJECTIVES: The goal of this study was to use an acute synovitis inflammation model to determine the residual effects of IA betamethasone and triamcinolone acetonide on various inflammatory parameters and lameness. STUDY DESIGN: Crossover randomised trial. METHODS: Five mixed-breed, 2-year-old horses were randomly allocated to an IA treatment of the radiocarpal joint with 9 mg of either betamethasone or triamcinolone acetonide. Two weeks following treatment, horses were injected with 1 µg of lipopolysaccharide (LPS) diluted in 1 ml of saline. Following LPS injection, horses were crossed-over and both sets of injections were repeated after a washout period. Blood samples were collected at multiple time points for mRNA analysis, as well as serum amyloid A (SAA) and cortisol determination. At each time point, lameness was also subjectively scored. Additional injections with saline-only or LPS-only (twice) were conducted as negative and positive controls, respectively. Two-way repeated measures analysis of variance was used to analyse all data. RESULTS: Corticosteroid-only treatments result in significant mRNA expression differences, as well as significant and prolonged cortisol suppression. Following LPS injection, there was a residual treatment effect with triamcinolone evidenced by a significant treatment effect on IL-6 and PTGS1 (cyclooxygenase-1), lameness, SAA and cortisol concentrations, while only IL-6 expression was affected by betamethasone. MAIN LIMITATIONS: The acute synovitis model used here results in significant inflammation and is not representative of the low-grade inflammation seen with typical joint disease and residual anti-inflammatory effects may be more profound in naturally occurring joint disease. CONCLUSIONS: Current regulatory guidelines may be insufficient if the concern is residual anti-inflammatory effects. Additionally, intra-articular corticosteroid administration is not without risk, as evidenced by a significant suppression of serum cortisol concentration and, as such, the benefits of their administration should be weighed against those risks.


Asunto(s)
Enfermedades de los Caballos , Artropatías , Sinovitis , Caballos , Animales , Triamcinolona Acetonida/uso terapéutico , Betametasona/uso terapéutico , Hidrocortisona , Lipopolisacáridos , Cojera Animal/tratamiento farmacológico , Interleucina-6 , Sinovitis/inducido químicamente , Sinovitis/tratamiento farmacológico , Sinovitis/veterinaria , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Inflamación/veterinaria , Artropatías/veterinaria , Antiinflamatorios , Inyecciones Intraarticulares/veterinaria , Enfermedades de los Caballos/tratamiento farmacológico , Enfermedades de los Caballos/metabolismo
8.
Equine Vet J ; 55(5): 831-842, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-36273247

RESUMEN

BACKGROUND: Foals that develop pulmonary ultrasonographic lesions on Rhodococcus equi (R. equi) endemic farms are treated with antibiotics because those at risk of developing clinical pneumonia (~20%) cannot be recognised early. Candidate biomarkers identified using metabolomics may aid targeted treatment strategies against R. equi. OBJECTIVES: (1) To describe how foal ageing affects their plasma metabolome (birth to 8 weeks) and (2) to establish the effects that experimental infection with Rhodococcus equi (R. equi) has on foal metabolome. STUDY DESIGN: Experimental study. METHODS: Nine healthy newborn foals were experimentally infected with R. equi as described in a previous study. Foals were treated with oral antibiotics if they developed clinical pneumonia (n = 4, clinical group) or remained untreated if they showed no signs of disease (n = 5, subclinical group). A group of unchallenged foals (n = 4) was also included in the study. By the end of the study period (8 weeks), all foals were free of disease. This status was confirmed with transtracheal wash fluid evaluation and culture as well as thoracic ultrasonography. Plasma metabolomics was determined by GC-MS weekly for the study duration (8 weeks). RESULTS: Foals' plasma metabolome was altered by ageing (birth to 8 weeks) and experimental infection with R. equi as demonstrated using multivariate statistical analysis. The intensities of 25 and 28 metabolites were altered by ageing and infection (p < 0.05) respectively. Furthermore, 20 metabolites changed by more than 2-fold between clinical and subclinical groups. MAIN LIMITATIONS: The number of foals is limited. Foals were experimentally infected with R. equi. CONCLUSIONS: Ageing and R. equi infection induced changes in the plasma metabolome of foals. These results provide an initial description of foal's plasma metabolome and serve as background for future identification of R. equi pneumonia biomarkers.


INTRODUCTION/CONTEXTE: Les poulains qui développent des lésions pulmonaires échographiques dans les fermes d'élevage où Rhodococcus equi (R. equi) est endémique sont traités avec antibiotiques car ceux à risque de développer des lésions cliniques (~20%) ne peuvent être identifiés précocement. Certains biomarqueurs identifiés par le biais de la métabolomique pourraient aider à orienter les stratégies de traitement pour R. equi. OBJECTIFS: (1) Décrire les changements de métabolome plasmatique qui surviennent chez les poulains en lien avec l'âge (naissance jusqu'à 8 semaines d'âge) et (2) Établir les effets d'une infection expérimentale à Rhodococcus Equi sur le métabolome des poulains. TYPE D'ÉTUDE: Étude expérimentale. MÉTHODES: Neufs poulains nouveaux-nés en santé ont été infectés de façon expérimentale par R. equi tel que décrit précédemment. Ils ont été traités avec des antibiotiques s'ils ont développé une pneumonie clinique (n = 4, groupe clinique) ou ont simplement été suivi dans le temps s'ils n'ont pas montré de signes de la maladie (n = 5, groupe sous-clinique). Un groupe de poulains sains (n = 4) était aussi inclus dans l'étude. À la fin de l'étude (8 semaines), tous les poulains étaient sains tel que confirmé par l'évaluation et la culture de leur fluide de lavage transtrachéal de même qu'à l'échographie thoracique. Les métabolomiques plasmastiques ont été déterminées par GC-MS de façon hebdomadaire pour la durée de l'étude (8 semaines). RÉSULTATS: À la fois l'âge et l'infection expérimentale ont altéré le métabolome plasmatique des poulains tel que démontré par l'analyse statistique multivariée. L'âge a altéré l'intensité de 25 métabolites et l'infection a modifié l'intensité de 28 métabolites (p < 0.05). De plus, 20 métabolites ont changé de plus de 2 fois leur valeur initiale, entre les groupes cliniques et sous-cliniques. LIMITES PRINCIPALES: Le nombre de poulains reste limité. Les poulains ont été infecté par R. equi de façon expérimentale. CONCLUSIONS: Le vieillissement et l'infection par R. equi induisent des changements dans le métabolome plasmatique des poulains. Ces résultats représentent une description initiale du métabolome plasmatique chez le poulain et peuvent servir de base pour l'identification future de biomarqueurs pour la détection de pneumonie à Rhodococcus equi.


Asunto(s)
Infecciones por Actinomycetales , Enfermedades de los Caballos , Neumonía , Rhodococcus equi , Animales , Caballos , Infecciones por Actinomycetales/veterinaria , Enfermedades de los Caballos/epidemiología , Neumonía/veterinaria , Metaboloma , Antibacterianos
9.
PLoS One ; 18(9): e0290778, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37669266

RESUMEN

Neonates have different cellular composition in their bronchoalveolar lavage fluid (BALF) when compared to foals and adult horses; however, little is known about the non-cellular components of BALF. The objective of this study was to determine the proteomic composition of BALF in neonatal horses and to compare it to that of foals and adult horses. Bronchoalveolar lavage fluid samples of seven neonates (< 1 week age), four 5 to 7-week-old foals, and six adult horses were collected. Quantitative proteomics of the fluid was performed using tandem mass tag labeling followed by high resolution liquid chromatography tandem mass spectrometry and protein relative abundances were compared between groups using exact text. A total of 704 proteins were identified with gene ontology terms and were classified. Of these, 332 proteins were related to the immune system in neonates, foals, and adult horses. The most frequent molecular functions identified were binding and catalytic activity and the most common biological processes were cellular process, metabolic process, and biological regulation. There was a significant difference in the proteome of neonates when compared to foals and to adult horses. Neonates had less relative expression (FDR < 0.01) of many immune-related proteins, including immunoglobulins, proteins involved in the complement cascade, ferritin, BPI fold-containing family B member 1, and macrophage receptor MARCO. This is the first report of equine neonate BALF proteomics and reveals differential abundance of proteins when compared to BALF from adult horses. The lower relative abundance of immune-related proteins in neonates could contribute to their susceptibility to pulmonary infections.


Asunto(s)
Líquidos Corporales , Proteómica , Caballos , Animales , Irrigación Terapéutica , Líquido del Lavado Bronquioalveolar , Cromatografía Liquida
10.
J Equine Vet Sci ; 109: 103828, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34843888

RESUMEN

The use of lipopolysaccharide to induce a localized source of inflammation (acute synovitis) and allow for monitoring of changes in systemic mRNA expression has been recently reported. Here, the goal was to maintain a significant systemic mRNA response while limiting the severity of lameness such that this model can be used to examine the effects of various anti-inflammatory treatment modalities on mRNA expression. Three mixed breeds, four-year-old geldings were utilized for this study. One milliliter of phosphate-buffered saline containing 1,000 ng or less of lipopolysaccharide from E. coli O111:B4 was aseptically injected into alternating radiocarpal joints following washout periods. Blood for complete blood cell count, serum amyloid A concentration, and mRNA analysis via RT-qPCR for 23 different genes were collected before each injection, as well as at multiple times post-injection. Lameness severity was also graded at each time point. Two-way, repeated measures analysis of variance was used for statistical analysis (P < .05). Results largely replicated those previously reported, with multiple genes exhibiting significant expression changes during the acute inflammatory period (including increases in CD14, TLR4, IL-1ß, IL1RN, MMP1, and MMP9 expression) while some demonstrated dose-dependent changes; significant increases in complete blood cell count parameters and serum amyloid A concentrations were also noted. Attempts to temper the severity of lameness were not successful as nonweight bearing lameness was noted at doses of 10ng or higher, while a dose of 1ng elicited neither a detectable lameness nor a significant change in mRNA expression.


Asunto(s)
Enfermedades de los Caballos , Sinovitis , Animales , Escherichia coli , Expresión Génica , Enfermedades de los Caballos/inducido químicamente , Caballos , Inyecciones Intraarticulares/veterinaria , Lipopolisacáridos , Masculino , Sinovitis/inducido químicamente , Sinovitis/veterinaria
11.
Equine Vet J ; 54(1): 121-131, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33445210

RESUMEN

BACKGROUND: Many foals that develop thoracic ultrasonographic lesions as a result of Rhodococcus equi infection heal on their own. However, most of these foals receive antimicrobials because foals at risk of developing clinical pneumonia cannot be identified. Untargeted lipidomics is useful to identify candidate biomarkers. OBJECTIVES: (a) To describe the changes that occur in foal lipidomics as a result of ageing (birth to 8 weeks) and (b) To compare these results with those observed in foals after experimental infection with R. equi. STUDY DESIGN: Experimental study. METHODS: Healthy newborn foals (n = 9) were challenged with R. equi intratracheally the first week of life. Foals were treated with antimicrobials if they developed clinical pneumonia (n = 4, "clinical group") or were closely monitored if they showed no signs of disease (n = 5 "subclinical group"). An unchallenged group (n = 4) was also included. All foals were free of disease (transtracheal wash fluid evaluation and culture as well as thoracic ultrasonography) by 8 weeks of life. Plasma lipidomics was determined by LC-MS weekly for the study duration (8 weeks). RESULTS: Both ageing and experimental infection altered the foal's plasma lipidome as demonstrated by multivariate statistical analysis. The intensities of 31 lipids were altered by ageing and 12 by infection (P < .05). Furthermore, nine lipids changed by more than twofold between clinical and subclinical groups. MAIN LIMITATIONS: The number of foals is limited. Foals were experimentally challenged with R. equi. CONCLUSIONS: Ageing and R. equi infection induced changes in the plasma lipidome of foals. These experimental results provide the background for future work in the discovery of earlier biomarkers of R. equi pneumonia. Early identification of foals at risk of developing clinical pneumonia is key in order to decrease antimicrobial use and development of antimicrobial resistance.


Asunto(s)
Infecciones por Actinomycetales , Enfermedades de los Caballos , Rhodococcus equi , Infecciones por Actinomycetales/tratamiento farmacológico , Infecciones por Actinomycetales/veterinaria , Animales , Antibacterianos , Caballos , Lipidómica
12.
J Equine Vet Sci ; 109: 103826, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34843887

RESUMEN

Cases of nocardioform placentitis are characterized by focal, mucoid placentitis resulting in late-term abortion, premature birth, or small, full-term foals, occur sporadically, and are most commonly associated with Crossiella equi and Amycolatopsis spp. infection. The goal of this project was to develop an enzyme-linked immunosorbent assay (ELISA) for quantifying antibodies against Crossiella equi and Amycolatopsis spp. and utilize the ELISA to determine when exposure occurs. Serum samples collected during the 2020 foaling season from Crossiella equi (n = 8) and Amycolatopsis spp. (n = 32) infected mares, as well as nonaffected mares (n = 51 mares), were used to develop and optimize bacteria-specific ELISAs. Following development of the ELISAs, banked serum samples from a single, central Kentucky Thoroughbred farm collected during 2012 to 2013 (n = 104 mares) and 2013-14 (n = 82 mares) were analyzed. Differences in various groups were analyzed using one-way analysis of variance (ANOVA). Crossiella equi-infected mares had significantly higher ELISA unit (EU) values on the Crossiella equi ELISA near parturition when compared to the other two groups (P < .001). Using the Amycolatopsis spp. ELISA, EU values were not significantly different between Amycolatopsis spp. infected and non-affected mares, suggesting this ELISA is not specific for Amycolatopsis spp. During 2013 to 2014, there were significant increases in EU values between June and late September for the Crossiella equi ELISA, suggesting exposure in the summer and early fall months. Data from the Crossiella equi ELISA may help provide a better understanding of the epidemiology of nocardioform placentitis, guide the development of a successful experimental challenge model, and allow for further refinement of these ELISAs.


Asunto(s)
Corioamnionitis , Enfermedades de los Caballos , Enfermedades Placentarias , Aborto Veterinario/epidemiología , Animales , Corioamnionitis/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Enfermedades de los Caballos/epidemiología , Caballos , Enfermedades Placentarias/epidemiología , Enfermedades Placentarias/veterinaria , Embarazo
13.
Equine Vet J ; 54(1): 63-73, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33438228

RESUMEN

BACKGROUND: The ability to identify horses at risk for catastrophic injuries continues to be a pressing issue for the racing industry, especially given recent events in North America. OBJECTIVES: Since most catastrophic injuries occur in areas of existing pathology and this pathology is likely to elicit an inflammatory response, it was hypothesised that analysis of messenger RNA (mRNA) expression would detect significant changes in select genes in horses at risk for a catastrophic injury. STUDY DESIGN: Prospective cohort study. METHODS: Five racing jurisdictions across the United States participated in this study. A total of 686 Tempus® RNA Blood Tube samples were collected for mRNA analysis from 107 catastrophically injured horses, as well as from noninjured horses sampled either prerace (n = 374) or postrace (n = 205). A subset of horses (n = 37) were sampled both prerace and postrace for analysis of expression changes during the postrace period. RESULTS: Of 21 genes analysed via RT-qPCR, the expression of 12 genes (ALOX5AP, CD14, IL-10, IL-1ß, IL-6, IL-8, MMP1, PTGS2, TLR4, TNFα, TNFSF13B and VEGFA) changed significantly within 45 minutes after a race and were excluded. Of the remaining nine genes (BMP-2, IGF-1, IL1RN, MMP2, MMP9, Osteoprotegrin, RANKL, SAA1 and TGFß), three genes (IGF-1, IL1RN and MMP2) were found to be significantly different between catastrophically injured and noninjured horses using multiple logistic regression modelling. Receiver operating characteristic analysis of models, which included mRNA expression, demonstrated sensitivities from 76%-82% (95% CI: 67%-93%) and specificities from 84%-88% (95% CI: 71%-94%) at the Youden Index. MAIN LIMITATIONS: Samples were collected as soon as possible postinjury (within 30 minutes). CONCLUSIONS: Analysis of mRNA expression of specific genes in the future may be considered as an economical, accessible and noninvasive means by which horses at risk for catastrophic injury can be identified.


Asunto(s)
ARN Mensajero , Animales , Caballos , Modelos Logísticos , América del Norte , Estudios Prospectivos , ARN Mensajero/genética , Factores de Riesgo
14.
Am J Vet Res ; 83(9)2022 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-35895773

RESUMEN

OBJECTIVE: To perform lipidomic analysis of surfactant and plasma from asthmatic and healthy horses. ANIMALS: 30 horses with clinical signs of asthma and 30 age-matched control horses. PROCEDURES: Detailed history, physical examination, CBC, and bronchoalveolar lavage fluid (BALF) cytologies were obtained. Asthmatic horses were grouped based on their BALF inflammatory profile: severe equine asthma (SEA), mild equine asthma with neutrophilic airway inflammation (MEA-N), or mild equine asthma with eosinophilic airway inflammation (MEA-E). Each asthma group was assigned its own age-matched control group. Lipidomic analysis was completed on surfactant and plasma. Surfactant protein D (SP-D) concentrations were measured in serum and BALF. RESULTS: SEA surfactant was characterized by a phospholipid deficit and altered composition (increased ceramides, decreased phosphatidylglycerol, and increased cyclic phosphatidic acid [cPA]). In comparison, MEA-N surfactant only had a decrease in select phosphatidylglycerol species and increased cPA levels. The plasma lipidomic profile was significantly different in all asthma groups compared to controls. Specifically, all groups had increased plasma phytoceramide. SEA horses had increased plasma cPA and diacylglycerol whereas MEA-N horses only had increased cPA. MEA-E horses had increases in select ceramides and dihydrocermides. Only SEA horses had significantly increased serum SP-D concentrations. CLINICAL RELEVANCE: The most significant surfactant alterations were present in SEA (altered phospholipid content and composition); only mild changes were observed in MEA-N horses. The plasma lipidomic profile was significantly altered in all groups of asthmatic horses and differed among groups. Data from a larger population of asthmatic horses are needed to assess implications for diagnosis, prognosis, and treatment.


Asunto(s)
Asma , Enfermedades de los Caballos , Surfactantes Pulmonares , Animales , Asma/diagnóstico , Asma/veterinaria , Líquido del Lavado Bronquioalveolar , Ceramidas , Enfermedades de los Caballos/metabolismo , Caballos , Inflamación/veterinaria , Lipidómica , Fosfatidilgliceroles , Fosfolípidos , Proteína D Asociada a Surfactante Pulmonar , Surfactantes Pulmonares/metabolismo , Tensoactivos
15.
J Virol ; 84(10): 4898-911, 2010 May.
Artículo en Inglés | MEDLINE | ID: mdl-20219931

RESUMEN

Extensive cell culture passage of the virulent Bucyrus (VB) strain of equine arteritis virus (EAV) to produce the modified live virus (MLV) vaccine strain has altered its tropism for equine CD3(+) T lymphocytes and CD14(+) monocytes. The VB strain primarily infects CD14(+) monocytes and a small subpopulation of CD3(+) T lymphocytes (predominantly CD4(+) T lymphocytes), as determined by dual-color flow cytometry. In contrast, the MLV vaccine strain has a significantly reduced ability to infect CD14(+) monocytes and has lost its capability to infect CD3(+) T lymphocytes. Using a panel of five recombinant chimeric viruses, we demonstrated that interactions among the GP2, GP3, GP4, GP5, and M envelope proteins play a major role in determining the CD14(+) monocyte tropism while the tropism for CD3(+) T lymphocytes is determined by the GP2, GP4, GP5, and M envelope proteins but not the GP3 protein. The data clearly suggest that there are intricate interactions among these envelope proteins that affect the binding of EAV to different cell receptors on CD3(+) T lymphocytes and CD14(+) monocytes. This study shows, for the first time, that CD3(+) T lymphocytes may play an important role in the pathogenesis of equine viral arteritis when horses are infected with the virulent strains of EAV.


Asunto(s)
Equartevirus/fisiología , Monocitos/virología , Mapeo de Interacción de Proteínas , Linfocitos T/virología , Proteínas del Envoltorio Viral/metabolismo , Tropismo Viral , Acoplamiento Viral , Animales , Complejo CD3/análisis , Células Cultivadas , Células Endoteliales/virología , Caballos , Receptores de Lipopolisacáridos/análisis , Monocitos/química , Linfocitos T/química
16.
Vet Res ; 42: 23, 2011 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-21314906

RESUMEN

Equine herpesvirus-1 (EHV-1) infection remains a significant problem despite the widespread use of vaccines. The inability to generate a protective immune response to EHV-1 vaccination or infection is thought to be due to immunomodulatory properties of the virus, and the ORF1 and ORF2 gene products have been hypothesized as potential candidates with immunoregulatory properties. A pony infection study was performed to define immune responses to EHV-1, and to determine if an EHV-1 ORF1/2 deletion mutant (ΔORF1/2) would have different disease and immunoregulatory effects compared to wild type EHV-1 (WT). Infection with either virus led to cytokine responses that coincided with the course of clinical disease, particularly the biphasic pyrexia, which correlates with respiratory disease and viremia, respectively. Similarly, both viruses caused suppression of proliferative T-cell responses on day 7 post infection (pi). The ΔORF1/ORF2 virus caused significantly shorter primary pyrexia and significantly reduced nasal shedding, and an attenuated decrease in PBMC IL-8 as well as increased Tbet responses compared to WT-infected ponies. In conclusion, our findings are (i) that infection of ponies with EHV-1 leads to modulation of immune responses, which are correlated with disease pathogenesis, and (ii) that the ORF1/2 genes are of importance for disease outcome and modulation of cytokine responses.


Asunto(s)
Infecciones por Herpesviridae/veterinaria , Herpesvirus Équido 1/genética , Herpesvirus Équido 1/inmunología , Enfermedades de los Caballos/inmunología , Proteínas Virales/genética , Inmunidad Adaptativa , Animales , Anticuerpos Antivirales/sangre , Citocinas/sangre , Citocinas/genética , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Infecciones por Herpesviridae/inmunología , Infecciones por Herpesviridae/virología , Herpesvirus Équido 1/metabolismo , Enfermedades de los Caballos/virología , Caballos , Inmunidad Innata , Masculino , Mucosa Nasal/virología , ARN Mensajero/análisis , Distribución Aleatoria , Proteínas Virales/metabolismo , Viremia/inmunología , Viremia/veterinaria , Viremia/virología , Esparcimiento de Virus
17.
Vet Immunol Immunopathol ; 237: 110266, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-33991760

RESUMEN

BACKGROUND: Immunological mechanisms involved in the pathogenesis of mild to moderate equine asthma (MEA) are not completely understood. There are limited data on bronchoalveolar lavage fluid (BALF) and blood inflammatory cytokine profiles in racehorses with MEA, and the effect of racing on inflammatory cytokines is unknown. HYPOTHESIS/OBJECTIVES: We hypothesized that inflammatory cytokine gene expression in BALF and resting blood would be higher in racehorses with lower airway inflammation compared to healthy controls, and that gene expression in blood collected immediately post-race would be increased compared to resting blood in racehorses with lower airway inflammation. ANIMALS: 38 racing Thoroughbreds (samples: 30 resting blood, 22 post-race BALF, 41 post-race blood). METHODS: Prospective observational study. Inflammatory cytokine gene expression was determined in resting blood, post-race BALF and post-race blood from racehorses with lower airway inflammation and controls. RESULTS: Lower airway inflammation was diagnosed in 79 % of racehorses (23 % neutrophilic, 67 % mastocytic, and 10 % mixed). There was no difference in gene expression in BALF or resting blood between racehorses with lower airway inflammation and controls. IL-8 gene expression was higher in post-race blood compared to resting peripheral blood, regardless of disease (p = 0052). BALF neutrophil proportions increased with increasing IL-1ß gene expression in all sample types (p = 0.0025). BALF mast cell proportions increased with increasing TNF-α gene expression in post-race blood (p = 0.015). CONCLUSIONS AND CLINICAL IMPORTANCE: Lower airway inflammation was common in a population of racehorses without respiratory signs or exercise intolerance. Exercise alone increased peripheral blood IL-8 gene expression. Inflammatory cytokine gene expression was not increased in BALF or resting blood in horses with subclinical lower airway inflammation, precluding its diagnostic utility in clinical practice.


Asunto(s)
Asma Inducida por Ejercicio/veterinaria , Asma/veterinaria , Líquido del Lavado Bronquioalveolar/inmunología , Citocinas/genética , Enfermedades de los Caballos/genética , Inflamación/veterinaria , Animales , Asma/genética , Asma/inmunología , Asma/metabolismo , Asma Inducida por Ejercicio/genética , Asma Inducida por Ejercicio/inmunología , Líquido del Lavado Bronquioalveolar/citología , Expresión Génica , Enfermedades de los Caballos/metabolismo , Caballos , Inflamación/genética , Inflamación/inmunología , Mastocitos/inmunología , Neutrófilos/inmunología , Esfuerzo Físico/inmunología , Deportes
18.
Am J Vet Res ; 82(2): 152-157, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33480279

RESUMEN

OBJECTIVE: To evaluate surfactant protein D (SP-D) concentrations in serum and bronchoalveolar lavage fluid (BALF) from young healthy horses on pasture or housed in a typical barn. ANIMALS: 20 young healthy horses. PROCEDURES: Horses were randomly assigned to 1 of 2 groups (pasture, n = 10; barn, 10), and serum and BALF samples were collected for SP-D determination at baseline (all horses on pasture) and 2 weeks and 4 weeks after the barn group of horses was relocated from the pasture to the barn. Other evaluations included physical and tracheoscopic examinations. Findings were compared within and between groups. RESULTS: Physical and tracheoscopic examinations, CBC, and serum biochemical analysis did not reveal evidence of respiratory disease, and no significant differences were present within and between groups. Serum SP-D concentrations did not significantly differ within and between groups, but BALF SP-D concentrations were significantly lower for the barn group at 2 weeks but not at 4 weeks, compared with baseline. The BALF SP-D concentration-to-BALF total protein concentration ratio was < 1.5 and did not significantly differ within and between groups. CONCLUSIONS AND CLINICAL RELEVANCE: A mild decrease was evident in the concentration of SP-D in the BALF collected from young healthy horses after 2 weeks of exposure to a barn environment. The clinical importance of this finding remains to be determined.


Asunto(s)
Enfermedades de los Caballos , Enfermedades Respiratorias , Animales , Lavado Broncoalveolar/veterinaria , Líquido del Lavado Bronquioalveolar , Caballos , Proteína D Asociada a Surfactante Pulmonar , Enfermedades Respiratorias/veterinaria
19.
Viruses ; 13(7)2021 07 09.
Artículo en Inglés | MEDLINE | ID: mdl-34372536

RESUMEN

Equine rotavirus group A (ERVA) is one of the most common causes of foal diarrhea. Starting in February 2021, there was an increase in the frequency of severe watery to hemorrhagic diarrhea cases in neonatal foals in Central Kentucky. Diagnostic investigation of fecal samples failed to detect evidence of diarrhea-causing pathogens including ERVA. Based on Illumina-based metagenomic sequencing, we identified a novel equine rotavirus group B (ERVB) in fecal specimens from the affected foals in the absence of any other known enteric pathogens. Interestingly, the protein sequence of all 11 segments had greater than 96% identity with group B rotaviruses previously found in ruminants. Furthermore, phylogenetic analysis demonstrated clustering of the ERVB with group B rotaviruses of caprine and bovine strains from the USA. Subsequent analysis of 33 foal diarrheic samples by RT-qPCR identified 23 rotavirus B-positive cases (69.69%). These observations suggest that the ERVB originated from ruminants and was associated with outbreaks of neonatal foal diarrhea in the 2021 foaling season in Kentucky. Emergence of the ruminant-like group B rotavirus in foals clearly warrants further investigation due to the significant impact of the disease in neonatal foals and its economic impact on the equine industry.


Asunto(s)
Enfermedades de los Caballos/virología , Caballos/virología , Rotavirus/patogenicidad , Animales , Proteínas de la Cápside/genética , Diarrea/etiología , Diarrea/virología , Brotes de Enfermedades/veterinaria , Heces/virología , Kentucky , Filogenia , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Rotavirus/clasificación , Infecciones por Rotavirus/veterinaria
20.
Am J Reprod Immunol ; 86(1): e13396, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33569862

RESUMEN

PROBLEM: Minimal evidence exists supporting therapeutic selections for equine placentitis. The goal of this study was to characterize the anti-inflammatory effects of firocoxib when administered to mares with placentitis. METHODS: Mares (gestation D270-300) were assigned to: INFECT (n = 6; placentitis, no treatment), FIRO (n = 6; placentitis, firocoxib, 0.1 mg/kg, PO, daily), and NORM (n = 6; no infection/treatment). Allantoic fluid (8 hours, 24 hours, birth) and amniotic fluid (birth) were collected from mares after infection. Concentrations of IL-1ß, IL-6, TNF-α, IL-10, PGF2α , and PGE2 in fluids were measured by ELISA. mRNA expression of IL-1ß, IL-6, TNF-α, IL-8, IL-10, matrix metalloproteinases (MMPs) -1, 3, and 9 in fetal membranes/fetuses was quantified using real-time PCR. RESULTS: Allantoic TNF-α concentrations were lowest in FIRO at 8 hours and 24 hours post-infection; IL-6 concentrations were lower in FIRO than NORM at 8 hours, lower in FIRO than INFECT at 24 hours post-inoculation, and lower in NORM than FIRO or INFECT at birth. Marginal mean allantoic IL-ß and IL-10 concentrations were lower in FIRO and NORM than INFECT. Amniotic fluid cytokines were lowest in NORM with all measurements in that group being below the limit of detection. Allantoic PGF2α concentrations were lower in FIRO and INFECT than NORM at 8 hours post-inoculation, and lower in FIRO than INFECT or NORM at 24 hours post-inoculation. Allantoic PGE2 concentrations were lower in FIRO than INFECT. Amniotic PGF2α and PGE2 concentrations were lower in NORM than INFECT. In fetal membranes, group differences with respect to IL-1ß, IL-6, IL-8, and MMP1 were dependent on tissue type. CONCLUSIONS: Data suggest a suppressive effect of firocoxib administration on cytokine and prostaglandin production in mares with placentitis.


Asunto(s)
4-Butirolactona/análogos & derivados , Antiinflamatorios/uso terapéutico , Inhibidores de la Ciclooxigenasa 2/uso terapéutico , Enfermedades de los Caballos/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Enfermedades Placentarias/tratamiento farmacológico , Placenta/metabolismo , Sulfonas/uso terapéutico , 4-Butirolactona/uso terapéutico , Animales , Femenino , Caballos , Interleucina-6/metabolismo , Metaloproteinasa 1 de la Matriz/metabolismo , Placenta/patología , Embarazo , Prostaglandinas/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo
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