Immunological and pathological investigations in equine experimental uveitis

Background

The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist.

Objectives

The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings.

Methods

Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation.

Results

CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction.

Conclusions

Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

     

Cardiac evaluation of anesthetized Grevy's zebras (Equus grevyi)

OBJECTIVE: To determine ECG and echocardiographic measurements in healthy anesthetized Grevy's zebras (Equus grevyi). ANIMALS: 20 healthy zebras. PROCEDURES: Auscultation, base-apex ECG, and echocardiography were performed on anesthetized zebras. RESULTS: Low-grade systolic murmurs were detected in the left basilar region in 4 of 20 zebras. Evaluation of ECGs from 19 zebras revealed sinus rhythm with a predominantly negative QRS complex and a mean +/- SD heart rate of 67 +/- 10 beats/min. Echocardiograms of sufficient image quality were obtained for 16 zebras. Interventricular septal thickness in diastole, left ventricular chamber in diastole and systole, left atrial diameter, and left ventricular mass were significantly and moderately correlated with estimated body weight (r values ranged from 0.650 to 0.884). Detectable swirling of blood in the right and sometimes the left ventricles was detected in 9 of 16 zebras, whereas physiologic regurgitation of blood was detected for the aortic valve in 3 zebras, pulmonary valve in 2 zebras, mitral valve in 2 zebras, and tricuspid valve in 1 zebra. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study provide reference information for use in the cardiac evaluation of anesthetized Grevy's zebras.     

Response of healthy dogs to infusions of human serum albumin

OBJECTIVE: To evaluate the clinical and immunologic response in healthy dogs to infusions of human serum albumin (HSA). ANIMALS: 9 healthy purpose-bred mixed-breed dogs. PROCEDURES: Each dog was administered a 25% HSA solution once or twice. Various physical examination and laboratory variables were serially evaluated. Antibody against HSA was assayed before and after infusion by use of an ELISA. Intradermal testing was also conducted. A repeated-measures ANOVA or Friedman repeated-measures ANOVA on ranks was used to compare results for the variables. RESULTS: Adverse clinical reactions were observed after the first or second infusion in 3 dogs. Anaphylactoid reactions were observed in 1 of 9 dogs during the first infusion and in 2 of 2 dogs administered a second infusion. Two dogs developed severe edema and urticaria 6 or 7 days after an initial infusion. All dogs developed anti-HSA antibodies. Positive responses for ID tests were observed in 8 of 9 dogs. Short-term increases were detected in blood protein, total bilirubin, and calcium concentrations after HSA infusion. Serum cholesterol concentrations and platelet counts decreased after HSA infusion. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of HSA resulted in profound reactions in 2 of 9 dogs administered a single infusion and in 2 of 2 dogs administered a second infusion. This indicates that there is risk of life-threatening adverse reactions to HSA infusion in healthy dogs.     

In vitro effects of reactive oxygen metabolites, with and without flunixin meglumine, on equine colonic mucosa

OBJECTIVE: To determine effects of reactive oxygen metabolites (ROMs), with and without flunixin meglumine, on equine right ventral colon (RVC) in vitro. ANIMALS: 18 healthy horses and ponies. PROCEDURES: In 3 groups of 6 animals each, short-circuit current and conductance were measured in RVC mucosa in Ussing chambers. The 3 groups received physiologic saline (0.9% NaCl) solution, IV, 10 minutes before euthanasia and tissue incubation in Krebs-Ringer-bicarbonate (KRB) solution; flunixin meglumine (1.1 mg/kg, IV) 10 minutes before euthanasia and tissue incubation in KRB solution; or physiologic saline solution, IV, 10 minutes before euthanasia and incubation in KRB solution with 2.7 x 10(5)M flunixin meglumine. Incubation conditions included control (no addition) and ROM systems, including addition of 1 mM xanthine and 80 mU of xanthine oxidase (to produce the superoxide radical), 1 mM H(2)O(2), and 1 mM H(2)O(2) and 0.5 mM ferrous sulfate (to produce the hydroxyl radical). RESULTS: All ROMs that were added or generated significantly increased the short-circuit current except in tissues coincubated with flunixin meglumine, and they induced mild epithelial vacuolation and apoptosis, but did not disrupt the epithelium nor change conductance, lactate dehydrogenase release, or [(3)H]mannitol flux. CONCLUSIONS AND CLINICAL RELEVANCE: Responses to ROMs could be attributed to increased chloride secretion and inhibited neutral NaCl absorption in equine RVC, possibly by stimulating prostaglandin production. The ROMs examined under conditions of this study could play a role in prostaglandin-mediated colonic secretion in horses with enterocolitis without causing direct mucosal injury.     

Molecular and serologic characterization, pathogenicity, and protection studies with infectious bronchitis virus field isolates from California

In this study, we characterized three variant infectious bronchitis virus (IBV) strains isolated in 2003 and 2004 from broiler chickens in California and compared them to previously isolated California variant viruses and to common vaccine serotypes used in the United States. We conducted genetic, serologic, and pathogenicity studies on all three isolates, then tested different vaccines against one of the viruses. Genetically the three variant IBV strains, designated CA557/03, CA706/03, and CA1737/04, were not related to each other. GenBank BLAST database search and phylogenetic analysis of the hypervariable region of the S1 subunit of the spike gene to determine the most closely related viruses to the three variants showed the CA557/03 variant to be 81.8% similar to the CAV/CA56b/91 whereas the CA706/03 and CA1737/04 variant viruses were only distantly related to Dutch/D1466/81 (72.2%), a vaccine strain used in Europe, and Korea/K142/02 (72.7%), a Korean field isolate, respectively. Cross virus-neutralization testing showed that none of the 2003-04 California IBV variant viruses were serologically related to each other or to Ark, Conn, or Mass vaccine strains. In addition the CA1737/04 isolate was also tested against DE072 and found not to be serologically related. All three variant viruses were pathogenic in 1-wk-old broilers and vaccination with Mass/Conn followed by Holland/Conn provided 80% protection against the CA1737/04 virus. The 2003-04 California variant viruses were not compared with variants isolated in California during 1970s and 1980s because, to our knowledge, no genetic information is available and those viruses are no longer obtainable. This study shows that the CA557/03 virus was distantly related to the CAV-type viruses isolated in California in the early 1990s, but that none of the 2003-04 viruses were similar genetically or serologically to the CAL99-type viruses, indicating that new IBV variants continue to emerge and cause disease in commercial chickens in California.     

Effect of orally administered Lactobacillus casei on porcine reproductive and respiratory syndrome (PRRS) virus vaccination in pigs

The purpose of this study was to determine whether intranasal/oral administration of probiotics can assist vaccination efficacy against an important swine pathogen, porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV). A controlled challenge trial was performed employing: (a) pigs vaccinated against PRRS and treated with a Lactobacillus casei, (b) pigs vaccinated against PRRS only, (c) pigs treated with L. casei only, and (d) pigs neither vaccinated against PRRS nor treated with L. casei. All pigs were challenged intranasally with a wild PRRSV strain. There was no difference in clinical signs or rectal temperature among the four groups. However, pigs that received L. casei gained significantly more weight than pigs that did not. Vaccinated pigs did not gain more weight than nonvaccinated pigs. Vaccinated groups had significantly fewer viraemic pigs on days post-challenge 4, 11 and 17 than nonvaccinated groups of pigs. There was no effect of probiotic on prevalence or duration of viraemia. Among viraemic pigs, there was no significant difference in mean log base(10) titer of PRRS virus among groups. These results suggest that orally administered L. casei does not affect immune response in such a way as to affect PRRS viraemia or nasal shedding. However, it still appears to provide significant benefit when administered during vaccination as indicated by the higher bodyweight gain following PRRS virus infection.     

The clinical utility of the right lateral intercostal ultrasound scan technique in dogs

When performing abdominal ultrasonography in dogs, the right aspect of the liver, porta hepatis, right kidney, right adrenal gland, pancreas, and duodenum are often not fully visible from a ventral, or subcostal, approach. The right lateral intercostal plane is an alternative approach that allows evaluation of these structures. This report provides multiple case examples that demonstrate the sonographic anatomy via the right intercostal approach. Other cases are included to demonstrate indications for this approach. Animals in which the right intercostal approach may prove most useful include large- and giant-breed dogs; deep-chested dogs; dogs with gas distention of the stomach, duodenum, and colon; dogs with microhepatia; and those with abdominal effusion and pain.     

Treatment of severe immune-mediated thrombocytopenia with human IV immunoglobulin in 5 dogs

BACKGROUND: Glucocorticoids with or without other immunotherapy are the initial treatment of choice for dogs with severe immune-mediated thrombocytopenia (IMT). The majority of treated dogs will have improvements in platelet counts within 5 to 7 days of starting therapy, but complications from hemorrhage often occur before a response is seen. Human IV immunoglobulin (hIVIG) blocks Fc receptors on mononuclear phagocytic cells in dogs; it is used in people with idiopathic thrombocytopenic purpura. HYPOTHESIS: The purpose of this study was to describe adverse effects and benefit of hIVIG in addition to conventional immunosuppressive therapy in dogs with severe IMT. ANIMALS: Five client-owned dogs with severe primary IMT. METHODS: Case series. The hospital database was searched for dogs with primary IMT treated with hIVIG. RESULTS: No adverse effects were noted during or after hIVIG infusion in any treated dog. Over a 6-month follow-up, all dogs were clinically normal when using conventional immunosuppressive therapy. Human IVIG was administered 3 days after initiation of immunosuppressive therapy in 4 dogs, and, after 2 days, in 1 dog. In all dogs, the mean platelet counts pre- and 24 hours post-hIVIG infusion (0.28-0.76 g/kg) were 2,500/pL and 50,600/microL (62,750/microL for the 4 responders), respectively. One dog failed to respond as promptly to hIVIG (0.34 g/kg), and the platelet count increased to 66,000/microL after 9 days of immunosuppressive therapy. The mean duration of hospitalization post-hIVIG in all 5 dogs was 1.8 days (12 hours for responders), and the mean total length of hospitalization was 4.6 days (3.5 days for responders). Active hemorrhage resolved and no packed red blood cell transfusions were required after hIVIG infusion for responders. CONCLUSIONS AND CLINICAL IMPORTANCE: Human IVIG was well tolerated and appeared to be associated with rapid platelet count recovery and amelioration of clinical signs in most dogs with IMT.