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/microbiologíaRESUMEN
Intra-articular corticosteroids are a popular treatment choice for joint-associated pain and inflammation in horses despite recent work on the metabolic effects of these drugs. The goal of this project was to compare metabolic effects between intra-articular (IA) triamcinolone acetonide (TA) and an autologous protein solution (APS). Five mixed-breed geldings (4-9 years) were utilized for this project. Three identical and consecutive 28-day treatment blocks were used, with metacarpophalangeal IA treatments consisting of equal volumes of saline, a commercially available APS, or 9 mg of TA. Regular plasma and serum samples were collected for ACTH, cortisol, glucose, insulin, and thyroid hormone analysis, in addition to thyrotropin-releasing hormone (TRH) and oral sugar tests (OSTs). Significant treatment effects of IA TA were present at 48 h post-injection in both the TRH and the OST. There was also significant suppression by IA TA of baseline ACTH and cortisol between 2 h and 96 h post-treatment, hyperglycemia between 12 h and 48 h, and hyperinsulinemia at 32 h post-treatment. There were no treatment effects with respect to any measured thyroid hormones, nor were there any significant treatment effects of APS noted. Results suggest at least 2 days and up to 7 days should elapse between a single 9 mg IA TA treatment and OST and/or TRH testing. This study found that TA exhibits significant effects on ACTH, cortisol, glucose, and insulin, while the APS does not.
RESUMEN
Accumulating high-speed exercise has been identified as a significant risk factor for catastrophic injuries in racing Thoroughbreds. Injuries, regardless of severity, are a main cause of withdrawal from the racing industry, raising animal welfare concerns and resulting in significant economic losses. While most of the current literature focuses on injuries incurred during racing rather than training, the present study aims to help fill this gap. As such, peripheral blood was collected weekly, prior to exercise or administration of medication, from eighteen, two-year-old Thoroughbreds throughout their first season of race training. Messenger RNA (mRNA) was isolated and used to analyze the expression of 34 genes via RT-qPCR. Statistical analysis of the noninjured horses (n = 6) showed that 13 genes were significantly correlated with increasing average weekly high-speed furlong performance. Additionally, there was a negative correlation for CXCL1, IGFBP3, and MPO with both cumulative high-speed furlongs and week of training for all horses. Comparison of both groups showed opposing correlations between the anti-inflammatory index (IL1RN, IL-10, and PTGS1) and average weekly high-speed furlong performance. Furthermore, evaluation of training effects on mRNA expression during the weeks surrounding injury, showed differences between groups in IL-13 and MMP9 at -3 and -2 weeks prior to injury. While some previously reported relationships between exercise adaptation and mRNA expression were not noted in this study, this may have been due to the small sample size. Several novel correlations, however, were identified and warrant further investigation as markers of exercise adaptation or potential risk for injury.
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Estudios de Casos y Controles , Caballos , AnimalesRESUMEN
The use of dexamethasone to control equine asthma is a common and effective treatment. Although short-term systemic dexamethasone treatment has not been shown to induce systemic immunosuppression in the horse, the goal of this study was to determine whether inhaled ciclesonide, an FDA-approved drug for the treatment of equine asthma, exerts any systemic immunosuppressive effects when compared to dexamethasone-treated and untreated horses. Eighteen light, mixed breed horses, ranging in age from 3 to 8 years of age, were used for this study and randomly assigned to one of three treatment groups: (1) nontreated controls, (2) ciclesonide treatment, or (3) dexamethasone treatment. Blood was collected daily for steady-state messenger RNA (mRNA) analysis, as well as at Days 0, 5, 10, and 15 of treatment for in vitro stimulation with Concanavalin A (ConA). Messenger RNA relative quantities were determined using RT-qPCR for select genes. Two-way, repeated measures analysis of variance was used to analyze qPCR data and results considered significant at P < .05. There were significant decreases in the steady-state, whole-blood expression of granzyme B and interferon-γ due to dexamethasone treatment, when compared to the nontreated control group. Within ConA-stimulated samples, there remained a suppressive effect of dexamethasone treatment on granzyme B expression compared to nontreated control horses. Similar effects were not noted in the ciclesonide-treated horses. Significant effects of ciclesonide treatment on markers of immune function were not noted in this study, suggesting a low risk for immunosuppression with inhaled ciclesonide treatment.
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/veterinariaRESUMEN
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.
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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 , EmbarazoRESUMEN
BACKGROUND: A systemic and dysregulated immune response to infection contributes to morbidity and mortality associated with sepsis. Peripheral blood-derived mesenchymal stromal cells (PB-MSC) mitigate inflammation in animal models of sepsis. Allogeneic PB-MSC administered IV to horses is well-tolerated but therapeutic benefits are unknown. HYPOTHESIS: After IV lipopolysaccharide (LPS) infusion, horses treated with PB-MSC would have less severe clinical signs, clinicopathological abnormalities, inflammatory cytokine gene expression, and oxidative stress compared to controls administered a placebo. ANIMALS: Sixteen horses were included in this study. METHODS: A randomized placebo-controlled experimental trial was performed. Sixteen healthy horses were assigned to 1 of 2 treatment groups (1 × 109 PB-MSC or saline placebo). Treatments were administered 30 minutes after completion of LPS infusion of approximately 30 ng/kg. Clinical signs, clinicopathological variables, inflammatory cytokine gene expression, and oxidative stress markers were assessed at various time points over a 24-hour period. RESULTS: A predictable response to IV LPS infusion was observed in all horses. At the dose administered, there was no significant effect of PB-MSC on clinical signs, clinicopathological variables, or inflammatory cytokine gene expression at any time point. Antioxidant potential was not different between treatment groups, but intracellular ROS increased over time in the placebo group. Other variables that changed over time were likely due to effects of IV LPS infusion. CONCLUSIONS AND CLINICAL IMPORTANCE: Administration of allogeneic PB-MSC did not cause clinically detectable adverse effects in healthy horses. The dose of PB-MSC used here is unlikely to exert a beneficial effect in endotoxemic horses.
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Endotoxemia , Enfermedades de los Caballos , Células Madre Mesenquimatosas , Animales , Citocinas/genética , Citocinas/metabolismo , Endotoxemia/veterinaria , Enfermedades de los Caballos/tratamiento farmacológico , Caballos , Infusiones Intravenosas/veterinaria , Lipopolisacáridos , Células Madre Mesenquimatosas/metabolismoRESUMEN
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.
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ARN Mensajero , Animales , Caballos , Modelos Logísticos , América del Norte , Estudios Prospectivos , ARN Mensajero/genética , Factores de RiesgoRESUMEN
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.
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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 , TensoactivosRESUMEN
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.
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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 , DeportesRESUMEN
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.
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Enfermedades de los Caballos , Enfermedades Respiratorias , Animales , Lavado Broncoalveolar/veterinaria , Líquido del Lavado Bronquioalveolar , Caballos , Proteína D Asociada a Surfactante Pulmonar , Enfermedades Respiratorias/veterinariaRESUMEN
While the use of lipopolysaccharide (LPS) to induce inflammation has been well described in the horse, the object of this study was to evaluate the effect of repeated intra-articular LPS injections and determine whether this method may be of use to assess changes in gene expression related to inflammation. Six mixed breed horses were utilized for this study, with three horses aged 10-17 years (older group) and three horses aged 3 years (younger group). One milliliter of phosphate-buffered saline containing 3 µg of LPS from Escherichia coli O111:B4 was aseptically injected into either the radiocarpal or front fetlock joint a total of four times, with at least two weeks between each injection and a different joint injected each time. Serum for protein concentration quantification and whole blood for expression analysis of 20 different genes were collected before each injection, as well as at multiple times post-injection. Statistical analysis was performed using analysis of variance (one-way and two-way) (P < 0.05). All horses experienced minimal or non-weight bearing lameness at 4-6 hours post-LPS injection, which generally improved by 24 h and resolved by 48 h. Multiple genes exhibited significantly differential expression when compared to both the pre-injection and sham injection time points, including CD14, TLR4, MMP1, MMP9, IL-1ß, IL1RN, IL-10, ALOX5AP, IL-8, TNFα, CCL8, IGF1, and PTGS2. Additionally, multiple genes exhibited increased expression in horses where the radiocarpal joint was injected when compared to the fetlock joint, as well as in younger horses compared to older horses. Serum concentrations of serum amyloid A (SAA) were negative prior to injection while all horses demonstrated an increase by 9 h post-injection, which often remained until at least 144 h. Attempts to measure in vivo serum cytokine levels using a multiplex assay were not successful and believed to be due to the lower limits of detection for the assays. The measurement of mRNA expression of pro- and anti-inflammatory genes provide sensitive and rapid information regarding the inflammatory response to an acute, localized stimulus, although care must be taken when selecting target joints or age groups of horses as the transcriptional response may vary based on these choices.
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Expresión Génica , Inflamación/genética , Inflamación/veterinaria , Sinovitis/genética , Sinovitis/veterinaria , Animales , Células Sanguíneas/inmunología , Citocinas/sangre , Escherichia coli/química , Enfermedades de los Caballos , Caballos , Inflamación/sangre , Inyecciones Intraarticulares , Lipopolisacáridos , Masculino , Sinovitis/sangreRESUMEN
Previous work to evaluate various risk factors for failure to complete competitive endurance rides has examined clinicopathologic parameters, measurements of inflammation, and speed. Here, inflammatory markers were measured before, during, and after a long-distance, competitive endurance ride to examine the intraride dynamics of inflammatory marker expression and attempt to correlate those findings with whether a horse completed or failed to complete the ride. A total of 77 horses entered into the 2018 Tevis Cup Ride in California were enrolled in the study. Peripheral blood samples for mRNA isolation and gene expression analysis for ALOX5AP, CD14, IL-10, IL-1ß, IL-6, IL-8, MMP-1, TLR4, TNFα, and TNFSF13B were collected before, during (55 km and 110 km checkpoints), and after (160 km) the ride. No overall significant differences were found between groups of finishing and nonfinishing horses with regard to inflammatory marker expression. There were, however, time point-specific differences in mRNA expression, and, in some cases, these were group-specific. The overall pattern was a profound, initial increase in expression of inflammatory markers at the 55 km checkpoint. Some markers remained elevated beyond this point, whereas others began to decrease toward preride levels. While this work identified some similarities with previously published works, intraride sampling revealed additional changes in inflammatory marker expression. As such, investigators working with endurance horses should consider the addition of intraride sampling, when possible, to ensure that significant but short-lived changes in mRNA expression are not missed.
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Condicionamiento Físico Animal , Animales , Biomarcadores , Caballos , Resistencia Física , ARN Mensajero , Factores de RiesgoRESUMEN
In the horse, Lawsonia intracellularis infection results in equine proliferative enteropathy (EPE). While upwards of 100% of weanlings on an endemic farm may seroconvert, only a small percentage (approximately 5%) will develop clinical disease. Cell-mediated immune mechanisms likely play a role in resistance to L. intracellularis and the absence of a L. intracellularis-specific IFN-γ response has been associated with the development of EPE. The goal of this study was to determine whether protection from clinical EPE is associated with the induction of a systemic IgG sub-isotypic response consistent with a Th1-type cytokine response. To describe their L. intracellularis/EPE status, horses enrolled in this study were placed into one of three categories: seropositive-only, vaccinated, and presumptive clinical EPE. An existing ELISA method was modified to detect L. intracellularis-specific IgG(a), IgG(b), and IgG(t) antibodies using the mouse anti-equine hybridomas CVS-48, CVS-39, and CVS-40, respectively. Additionally, the existing ELISA method was used to quantify total IgG antibodies specific for L. intracellularis for comparison between the groups. Total L. intracellularis-specific IgG was found to be significantly higher (p<0.05) in presumptive clinical EPE cases (n=21) when compared with seropositive (exposed but unaffected) (n=36) and vaccinated horses (n=27). Further, a similar pattern for IgG(a) was seen in that the presumptive clinical EPE horses had significantly more L. intracellularis-specific IgG(a) (p<0.05) than the seropositive or vaccinated horses. With IgG(b), however, the vaccinated horses had significantly more IgG(b) (p<0.05) than the presumptive clinical or seropositive horses. No L. intracellularis-specific IgG(t) was detected in samples from any of the groups. While the results presented here with respect to IgG(a) response in the presumptive clinical EPE group were expected, a higher concentration of IgG(a) was anticipated in the seropositive horses that failed to develop clinical disease as well as in the vaccinated horses. Future work utilizing newer reagents against a broader range of equine IgG sub-isotypes may provide additional information once these become commercially available.
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Infecciones por Desulfovibrionaceae/veterinaria , Enfermedades de los Caballos/microbiología , Inmunoglobulina G/inmunología , Enfermedades Intestinales/veterinaria , Lawsonia (Bacteria)/inmunología , Animales , Infecciones por Desulfovibrionaceae/inmunología , Infecciones por Desulfovibrionaceae/microbiología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Enfermedades de los Caballos/inmunología , Caballos , Enfermedades Intestinales/inmunología , Enfermedades Intestinales/microbiología , Isoformas de Proteínas/inmunologíaRESUMEN
The addition of streptolysin-O (SLO) to the standard antibiotics regimen was shown to be superior to antibiotics alone after experimental infection of foals with Rhodoccocus equi (R. equi). The objective of this study is to investigate this response by determining the site-specific expression of extracellular matrix (ECM) and inflammatory response genes in biopsy samples taken from three distinct lung regions of the infected foals. Twenty-four foals were challenged by intrabronchial instillation of R. equi and assigned to four treatment groups: SLO/antibiotics adjunct therapy, antibiotics-only therapy (7.5 mg/kg clarithromycin and 5 mg/kg rifampin), SLO-only, and saline-only treatments. Treatments were administered twice daily for 16 days unless symptoms progressed to the point where the foals needed to be euthanized. Gene expressions were determined using custom-designed equine real-time qPCR arrays containing forty-eight genes from ECM remodeling and inflammation pathways. A non-parametric Wilcoxon signed-rank test for independent samples was applied to two pairs of time-matched comparison groups, SLO/antibiotics vs. antibiotics-only and SLO-only vs. saline-only, to document the significant differences in gene expressions within these groups. Several genes, MMP9, MMP2, TIMP2, COL1A1, COL12A1, ITGAL, ITGB1, FN1, CCL2, CCL3, CXCL9, TNFα, SMAD7, CD40, IL10, TGFB1, and TLR2, were significantly regulated compared to the unchallenged/untreated control foals. The results of this study demonstrate that enhancement of clinical responses by SLO is consistent with the changes in expression of critical genes in ECM remodeling and inflammatory response pathways.
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Infecciones por Actinomycetales/veterinaria , Enfermedades de los Caballos/tratamiento farmacológico , Enfermedades Pulmonares/veterinaria , Rhodococcus equi/aislamiento & purificación , Estreptolisinas/farmacología , Infecciones por Actinomycetales/tratamiento farmacológico , Infecciones por Actinomycetales/inmunología , Infecciones por Actinomycetales/microbiología , Animales , Proteínas Bacterianas/farmacología , Biopsia/veterinaria , Matriz Extracelular/genética , Matriz Extracelular/inmunología , Enfermedades de los Caballos/genética , Enfermedades de los Caballos/microbiología , Caballos , Enfermedades Pulmonares/tratamiento farmacológico , Enfermedades Pulmonares/genética , Enfermedades Pulmonares/inmunología , Enfermedades Pulmonares/microbiología , ARN Bacteriano/química , ARN Bacteriano/genética , Distribución Aleatoria , Reacción en Cadena en Tiempo Real de la Polimerasa , Estadísticas no ParamétricasRESUMEN
Equine influenza A (H3N8) virus infection is a leading cause of respiratory disease in horses, resulting in widespread morbidity and economic losses. As with influenza in other species, equine influenza strains continuously mutate, often requiring the development of new vaccines. Current inactivated (killed) vaccines, while efficacious, only offer limited protection against diverse subtypes and require frequent boosts. Research into new vaccine technologies, including gene-based vaccines, aims to increase the neutralization potency, breadth, and duration of protective immunity. Here, we demonstrate that a DNA vaccine expressing the hemagglutinin protein of equine H3N8 influenza virus generates homologous and heterologous immune responses, and protects against clinical disease and viral replication by homologous H3N8 virus in horses. Furthermore, we demonstrate that needle-free delivery is as efficient and effective as conventional parenteral injection using a needle and syringe. These findings suggest that DNA vaccines offer a safe, effective, and promising alternative approach for veterinary vaccines against equine influenza.
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Enfermedades de los Caballos/prevención & control , Subtipo H3N8 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Vacunación/métodos , Vacunas de ADN/inmunología , Animales , Anticuerpos Antivirales/sangre , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Caballos , Subtipo H3N8 del Virus de la Influenza A/genética , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/efectos adversos , Ratones , Infecciones por Orthomyxoviridae/prevención & control , Vacunación/efectos adversos , Vacunas de ADN/administración & dosificación , Vacunas de ADN/efectos adversosRESUMEN
While interferon-gamma (IFNγ) plays an important role in protection against viral and intracellular bacterial infections, its production in neonates is deficient. Exposure to environmental antigens can promote the maturation of the immune system of neonatal humans and mice. We hypothesize that exposure to high level of microbial components would increase the production of IFNγ in neonatal foals. To test this hypothesis, one group of foals was placed into stalls three times a week for 8 weeks. A second group of foals remained on pasture. Air samples were collected from the barn and pasture for microbial culture. There were more bacteria and fungi in the air samples collected from the barn compared with those from the pasture. Bronchoalveolar lavage (BAL) cells and peripheral blood mononuclear cells (PBMC) were collected from both groups of foals at various times to assess IFNγ production. The frequency of IFNγ(+) lymphocytes in BAL cells and PBMC was higher for foals kept in the stalls.
Asunto(s)
Caballos/inmunología , Interferón gamma/biosíntesis , Microbiología del Aire , Animales , Animales Recién Nacidos , Bacterias/inmunología , Bacterias/aislamiento & purificación , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Microbiología Ambiental , Hongos/inmunología , Hongos/aislamiento & purificación , Caballos/microbiología , Sistema Inmunológico/crecimiento & desarrollo , Leucocitos Mononucleares/inmunologíaRESUMEN
OBJECTIVE: To assess the serial use of serum immunoperoxidase monolayer assays (IPMAs) and fecal PCR assays, combined with other diagnostic methods, to identify subclinical Lawsonia intracellularis infections for targeted treatment of Thoroughbred foals and weanlings at farms in which the pathogen was endemic or nonendemic. DESIGN: Evaluation study. ANIMALS: 100 foals and weanlings (53 and 47 at farms in which L intracellularis was endemic and nonendemic, respectively). PROCEDURES: Serum was collected every 4 weeks and tested via IPMA, for antibodies against L intracellularis. Fecal samples were collected every 2 weeks and tested by use of an L intracellularis-specific PCR assay. When results for IPMAs or PCR assays were positive or clinical signs compatible with equine proliferative enteropathy (EPE) were detected, clinicopathologic testing was performed to determine treatment. RESULTS: No foals had positive results for the L intracellularis-specific IPMA until after weaning; 32 of 53 (60.4%) weanlings at the farm in which L intracellularis was endemic and 8 of 47 (170%) at the farm in which L intracellularis was nonendemic had positive IPMA results, whereas the number of weanlings that tested positive via fecal PCR assays at those farms was 6 and 0, respectively. Nineteen of 32 weanlings with positive IPMA results at the farm in which L intracellularis was endemic were treated for EPE; 5 of these had clinical signs of EPE. No weanlings at the nonendemic farm had clinical signs of or were treated for EPE. CONCLUSIONS AND CLINICAL RELEVANCE: IPMA appeared to be a useful means of identifying weanlings exposed to L intracellularis.
Asunto(s)
Infecciones por Desulfovibrionaceae/veterinaria , Enfermedades de los Caballos/diagnóstico , Lawsonia (Bacteria) , Reacción en Cadena de la Polimerasa/veterinaria , Pruebas Serológicas/veterinaria , Animales , Animales Recién Nacidos , Anticuerpos Antibacterianos/sangre , Infecciones por Desulfovibrionaceae/diagnóstico , Diagnóstico Diferencial , Heces/microbiología , Femenino , Caballos , Lawsonia (Bacteria)/inmunología , Lawsonia (Bacteria)/aislamiento & purificación , MasculinoRESUMEN
Rhodococcus equi is a soil borne bacterium that causes severe morbidity and death in young foals. The economic costs of the disease include loss of life, treatment expenses, veterinary monitoring expenses and, perhaps most importantly, potential reduction in future athletic performance in horses that suffer severe lung abscessations caused by R. equi. Current standard of care for pneumonia caused by R. equi is treatment with a macrolide antimicrobial and rifampicin. However, the hallmark of pneumonia caused by R. equi is severe formation of pyogranulomas and a walling off effect that can prevent systemic antibiotics from reaching antimicrobial concentrations in lung tissues. It is hypothesized that streptolysin O (SLO) used as an adjunct therapy with antibiotics will reduce the duration and severity of disease caused by R. equi pneumonia compared to antibiotic therapy alone. Addition of SLO to the antibiotic enhanced clinical responses compared to the other groups, including the antibiotic alone group. Of particular significance were lower bacterial counts in the lungs and longer survival time in those foals treated with SLO and antibiotics.