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1.
Horses inoculated with either equine cornea or killed Leptospira interrogans serovars pomona, tarassovi, icterohaemorrhagiae, wolffi and hardjo, developed corneal opacity and produced antibodies which made it possible to demonstrate partial antigenic identity between equine cornea and four of those serovars employed. These antibodies were isolated by means of immunoadsorptions, purified by ion-exchange chromatography (DEAE-Sephadex A-50) and run by immuno-electrophoresis in agar gel. Both antibodies, anti-equine cornea and anti-leptospira, showed that they corresponded to the IgGb subclass. They bound themselves to equine cornea in vivo and in vitro as was proved by immunofluorescence. This antigenic relationship may be in part responsible for pathogenesis of corneal opacity in leptospirosis of horses.  相似文献   

2.
A quantitative investigation of equine tear and aqueous humor immunoglobulins was done using normal horses and ponies as well as horses and ponies infected with Onchocerca cervicalis. The equine immunoglobulin isotypes IgGa, IgM, IgA and IgG(T) were quantitated by either single radial immunodiffusion (SRID) or radioimmunoassay (RIA). Tear immunoglobulin levels for IgGa (128 +/- 151 micrograms/ml), IgA (1,664 +/- 1,038 micrograms/ml) and IgM (106 +/- 74 micrograms/ml) were measured, while IgG(T) was not detectable. In horses with ocular inflammation the IgGa was 18-fold higher in the tears, 2,269 +/- 3,077 micrograms/ml. Aqueous humor obtained by paracentesis of the normal equine eye under anaesthesia, resulted in values for IgGa (45.2 +/- 20.0 micrograms/ml), IgG(T) (5.2 +/- 2.0 micrograms/ml), IgM (1.3 +/- 4.8 micrograms/ml) and IgA (0.8 +/- 1.0 micrograms/ml). A pooled sample of normal aqueous fluid obtained from over 100 horses at an equine abbatoir in Indiana gave values of 1,150 micrograms/ml for IgGa, 65 micrograms/ml for IgG(T), 2.5 micrograms/ml for IgA and 3.0 micrograms/ml for IgM. In animals infected with 0. cervicalis and treated with Diethylcarbamazine (DEC), there was a marked elevation of IgGa and IgG(T) in the tears and aqueous humor while IgA and IgG(T) were also elevated slightly in the aqueous. The findings of elevated immunoglobulin isotypes in the aqueous humor may not be related to the DEC treatment and 0. cervicalis infections but rather to repeated paracentesis and the development of acute inflammation of the equine eye as a result of the trauma of paracentesis. The elevations in equine immunoglobulin isotypes in the tears after DEC treatment are not subject to the same caveat. The preferential elevation in IgGa and IgG(T) in the tears precedes the development of corneal opacities observed in the same horses. The concentration of specific antimicrofilarial antibody in these tears remains to be determined but may well account for a major share of the total immunoglobulins detected.  相似文献   

3.
OBJECTIVE: To determine the role of intraocular bacteria in the pathogenesis of equine recurrent uveitis (ERU) in horses from the southeastern United States by evaluating affected eyes of horses with ERU for bacterial DNA and intraocular production of antibodies against Leptospira spp. SAMPLE POPULATION: Aqueous humor, vitreous humor, and serum samples of 24 clinically normal horses, 52 horses with ERU, and 17 horses with ocular inflammation not associated with ERU (ie, non-ERU inflammation). PROCEDURES: Ribosomal RNA quantitative PCR (real-time PCR) assay was used to detect bacterial DNA in aqueous humor and vitreous humor from clinically normal horses (n = 12) and horses with chronic (> 3-month) ERU (28). Aqueous humor and serum were also evaluated for anti-Leptospira antibody titers from clinically normal horses (n = 12), horses with non-ERU inflammation (17), and horses with confirmed chronic ERU (24). RESULTS: Bacterial DNA was not detected in aqueous humor or vitreous humor of horses with ERU or clinically normal horses. No significant difference was found in titers of anti-Leptospira antibodies in serum or aqueous humor among these 3 groups. Only 2 horses, 1 horse with ERU and 1 horse with non-ERU inflammation, had definitive intraocular production of antibodies against Leptospira organisms. CONCLUSIONS AND CLINICAL RELEVANCE: In horses from the southeastern United States, Leptospira organisms may have helped initiate ERU in some, but the continued presence of the organisms did not play a direct role in the pathogenesis of this recurrent disease.  相似文献   

4.
The antigenic relationship between Leptospira interrogans, equine cornea and lens was previously noted in our studies. Serum antibodies from horses inoculated with serovars wolffi, pomona, icterohaemorrhagiae, and tarassovi, were able to bind to five antigenic fractions from both cornea and lens, as demonstrated by immunoblotting. These antigens seem to be made up of protein and carbohydrates. After treatment with periodate for cleavage of glycoside ring structures, those fractions kept their condition of target for anti-Leptospira antibodies. Nevertheless, all fractions lost that condition after being subjected to peptide hydrolysis.  相似文献   

5.
An indirect hemagglutination was developed for the diagnosis of equine infectious anemia using sheep red blood cells coated with group specific virus antigen which had been highly purified by affinity chromatography. The presence of indirect hemagglutination antibodies was demonstrated in horses with equine infectious anemia since the cells were specifically agglutinated by all the serum samples obtained from experimentally infected horses. Antibodies appeared within 35 days after inoculation, and development of which coincided well with that of precipitating and complement fixing antibodies. Titer of indirect hemagglutination antibodies were ten to 320 times greater than those of precipitating antibodies. Test results could be read more clearly by the indirect hemagglutination test especially in weakly positive cases. Ninety-six samples from suspected field cases collected from every region of Japan which were positive on the immunodiffusion test were also positive on indirect hemagglutination test. Serum samples from 420 horses in one race track were examined by both the indirect hemagglutination and immunodiffusion tests to determine the reliability of the indirect hemagglutination test for diagnosis of equine infectious anemia. The same result was obtained on both tests. Based on this evidence, the indirect hemagglutination test can be employed as a very sensitive serological test for the diagnosis of equine infectious anemia.  相似文献   

6.
Equine recurrent uveitis (ERU) is the most frequent cause of blindness in horses worldwide. Leptospira has been implicated as an etiologic agent in some cases of ERU and has been detected in fresh ocular tissues of affected horses. The objective of this study was to determine the presence of Leptospira antigen and DNA in fixed equine ocular tissues affected with end-stage ERU. Sections of eyes from 30 horses were obtained. Controls included 1) 10 normal equine eyes and 2) 10 equine eyes with a nonrecurrent form of uveitis. The experimental group consisted of 10 eyes diagnosed with ERU based on clinical signs and histologic lesions. Sections were subjected to immunohistochemical staining with an array of rabbit anti-Leptospira polyclonal antibodies. DNA extractions were performed by using a commercial kit designed for fixed tissue. Real-time PCR analysis was completed on extracted DNA. The target sequence for PCR was designed from alignments of available Leptospira 16S rDNA partial sequences obtained from GenBank. Two of 10 test samples were positive for Leptospira antigen by immunohistochemical assay. Zero of 20 controls were positive for Leptospira antigen. All test samples and controls were negative for Leptospira DNA by real-time PCR analysis. Leptospira was detected at a lower frequency than that previously reported for fresh ERU-affected aqueous humor and vitreous samples. Leptospira is not frequently detectable in fixed ocular tissues of horses affected with ERU when using traditional immunohistochemical and real-time PCR techniques.  相似文献   

7.
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8.
In horses, allergies have been characterized by clinical signs and/or intradermal (i.d.) allergen testing. Our aim was to find the first direct evidence that immunoglobulin E (IgE) mediates equine allergy. In addition, we tested the hypothesis that immediate skin reactions in horses can also be mediated by IgG. Anti-IgE affinity columns were used to purify IgE from serum of one healthy horse and three horses affected with summer eczema, an allergic dermatitis which is believed to be induced by Culicoides midges. A modified Prausnitz-Küstner experiment was performed in four clinical healthy horses by i.d. injection of the purified serum IgE antibodies. The following day, Culicoides allergen was injected at the same sites. Skin reactions were not observed in response to allergen alone, and in two horses after stimulation at any previous IgE injection site. However, the other two horses showed an immediate skin reaction at the previous injection sites of IgE obtained from allergic horses. In addition, purified monoclonal antibodies to various equine immunoglobulin isotypes were injected i.d. into six healthy horses. Immediate skin reactions were observed in response to anti-IgE (6/6 horses) and anti-IgG(T) injections (5/6 horses). The specificities of both antibodies for IgE and IgG(T), respectively, were confirmed by enzyme linked immunosorbent assays. The results provide the first direct evidence that IgE mediates classical Type-I allergy in horses and plays a major role in the pathogenesis of summer eczema. The data also suggest that IgG(T) can bind to skin mast cells and might contribute to clinical allergy.  相似文献   

9.
OBJECTIVE: To evaluate expression of cyclooxygenase (COX)-1 and COX-2 in the cornea, eyelid, and third eyelid of healthy horses and those affected with squamous cell carcinoma (SCC) by use of immunohistochemical techniques. ANIMALS: 15 horses with SCC involving ocular tissues and 5 unaffected control horses. PROCEDURES: SCC-affected tissues were obtained from the cornea (n = 5 horses), eyelid (5), and third eyelid (5). Site-matched control tissues were obtained from 5 horses unaffected with SCC. Tissue sections of affected and control cornea, eyelid, and third eyelid were stained immunohistochemically for COX-1 and COX-2 via standard techniques. Stain uptake was quantified by use of computer-assisted image analysis of digital photomicrographs. RESULTS: Immunoreactivity for both COX-1 and COX-2 was significantly greater in equine corneas with SCC than in control corneas. No significant differences in COX-1 or COX-2 immunoreactivity were detected in eyelid and third-eyelid SCC, compared with site-matched control tissues. CONCLUSIONS AND CLINICAL RELEVANCE: Immunoreactivity for COX-1 and COX-2 is high in equine corneal SCC, possibly indicating that COX plays a role in oncogenesis or progression of this tumor type at this site. Pharmacologic inhibition of COX may represent a useful adjunctive treatment for corneal SCC in horses.  相似文献   

10.
Immunoglobulin E forms a minor component of serum antibody in mammals. In tissues IgE is bound by FcvarepsilonRI receptors on the surface of mast cells and mediates their release of inflammatory substances in response to antigen. IgE and mast cells have a central role in immunity to parasites and the pathogenesis of allergic diseases in horses and other mammals. This paper describes the production of several novel monoclonal antibodies that detect native equine IgE in immunohistology, ELISA and Western blotting. An antigen capture ELISA to quantify equine IgE in serum has been developed using two of these antibodies. The mean serum IgE concentration of a group of 122 adult horses was 23,523ng/ml with a range of 425-82,610ng/ml. Total serum IgE of healthy horses was compared with that of horses with insect bite dermal hypersensitivity (IBDH) an allergic reaction to the bites of blood feeding insects of Culicoides or Simulium spp. IBDH does not occur in Iceland where Culicoides spp. are absent, but following importation into mainland Europe native Icelandic horses have an exceptionally high incidence of this condition. In the present study Icelandic horses with IBDH had significantly higher total IgE than healthy Icelandic horse controls (P<0.05). By contrast in horses of other breeds the difference in total serum IgE between those affected with IBDH and healthy controls was not statistically significant. Total serum IgE was also monitored in a cohort of Icelandic horses prior to import into Switzerland and for a period of 3 years thereafter. High levels of serum IgE were present in all horses at the start of the study but dropped in the first year after import. Thereafter the total serum IgE remained low in Icelandic horses that remained healthy but rose significantly (P<0.05) in those that developed IBDH. These results support the conclusion that IBDH is a type I hypersensitivity response to insect allergens but indicate that IBDH in Icelandic horses may have a different pathogenesis from the same condition in other breeds.  相似文献   

11.
Summary The prevalence of antibodies to various viruses was investigated in a series of serum samples collected from horses in the Netherlands between 1963 and 1966 and from 1972 onwards. Neutralizing antibodies to equine rhinopneumonitis virus, equine arteritis virus and to equine rhinovirus types 1 and 2 were detected in respectively 76%, 14%, 66% and 59% of the equine serum samples tested. The observed incidence of serum samples positive to equine adenovirus in the complement fixation test was 39%. Precipitating antibodies to equine infectious anaemia virus were detected only in serum samples from two horses imported from abroad. Haemagglutination inhibiting antibodies to Myxovirus influenzae A / equi-1, M. Influenzae A / equi-2, and Reovirus types 1, 2, and 3 were present in respectively 82%, 50%, 10%, 33% and 3.6% of the serum samples tested. The most frequently observed incidence of antibodies to the various equine respiratory viruses occurred in the groups of horses having repeatedly contact with other horses.  相似文献   

12.
Background: Little information is available about experimental induction of leptospirosis in horses. Objectives: Determine serologic, hematologic responses of horses to Leptospira interrogans serovar Kennewicki infection. Animals: Four adult horses seronegative for leptospirosis. Methods: Experimental and observational study. Horses were challenged with an equine isolate of L. interrogans serovar Kennewicki at 2 different doses and different inoculation sites. After challenge, the horses were monitored for 60 days. Blood, urine, and aqueous humor samples were collected at intervals until euthanasia 60 days after infection. Results: Pyrexia (39.3–40°C) occurred as early as 1 day after challenge with 10 × 108Leptospira divided equally between topical ocular and intraperitoneal injection in 2 horses. Leptospires were recovered from the blood and urine but not from the aqueous humor of the 2 febrile horses. The sera of all 4 challenged horses developed microscopic agglutination test antibody after challenge and remained relatively constant for 21 days. Titer to cross‐reacting strains declined earlier than titer to the challenge strain. Conclusions: Clinical disease in experimentally infected horses can be mild or inapparent in Leptospira infected horses. Repeated serologic testing can allow recognition of the infecting serovar. In febrile horses, Leptospira can be isolated from blood while isolation from the urine can occur after fever has subsided.  相似文献   

13.
Summary

The prevalence of antibodies to various viruses was investigated in a series of serum samples collected from horses in the Netherlands between 1963 and 1966 and from 1972 onwards. Neutralizing antibodies to equine rhinopneumonitis virus, equine arteritis virus and to equine rhinovirus types 1 and 2 were detected in respectively 76%, 14%, 66% and 59% of the equine serum samples tested.

The observed incidence of serum samples positive to equine adenovirus in the complement fixation test was 39%. Precipitating antibodies to equine infectious anaemia virus were detected only in serum samples from two horses imported from abroad. Haemagglutination inhibiting antibodies to Myxovirus influenzae A / equi‐1, M. Influenzae A / equi‐2, and Reovirus types 1, 2, and 3 were present in respectively 82%, 50%, 10%, 33% and 3.6% of the serum samples tested.

The most frequently observed incidence of antibodies to the various equine respiratory viruses occurred in the groups of horses having repeatedly contact with other horses.  相似文献   

14.
C3 was detected bound in vivo to the opaque cornea of horses inoculated with killed Leptospira interrogans. Employing epithelial corneal cells isolated from a monolayer in tissue culture, we proved that C3 is fixed in vitro to the intact cell surface after incubation with a fresh equine anti-Leptospira serum. These findings, in addition to the infiltration of cornea with neutrophils and lymphocytes, may explain the mechanisms of tissue damage in recurrent uveitis of horses with leptospirosis.  相似文献   

15.
A 6-year-old Quarter Horse stallion was referred to Oklahoma State University Veterinary Medical Teaching Hospital for evaluation of abdominal pain that developed after breeding activity earlier in the day. The horse developed diarrhea and progressively worsening neurologic signs (circling, ataxia, head pressing) within 22 hours of presentation and was subsequently euthanized due to severe self-destructive behavior. Antemortem biochemical and hematologic abnormalities included hypocalcemia but no evidence of hepatic disease. Idiopathic hyperammonemia and encephalopathy were suspected; cerebrospinal fluid (CSF) and aqueous humor were collected 10 hours postmortem for ammonia analysis using a colorimetric assay. Results were compared with those of 6 horses that also had been euthanized, for diseases unrelated to encephalopathy. Ammonia also was measured in plasma samples obtained antemortem. Ammonia concentrations in plasma (958 micromol/L), CSF (1566 micromol/L) and aqueous humor (1018 micromol/L) samples from the stallion were markedly increased compared to those in the 6 unaffected horses (plasma, 9-43 micromol/L; CSF, 370-532 micromol/L; aqueous humor, 70-483 micromol/L). Since the acute nature of hyperammonemic encephalopathy often does not provide sufficient time for an antemortem diagnosis, postmortem analysis of CSF and aqueous humor ammonia concentrations may be a useful alternative for documenting hyperammonemia in horses.  相似文献   

16.
Lameness due to stifle and especially meniscal lesions is frequent in equine species. In humans, mechanoreceptors involved in proprioceptive function are well studied. Given the high incidence of meniscal injuries in horses, and the lack of information concerning them in equine menisci, our objective was to study these corpuscles in six healthy anterior horns of the equine medial meniscus, which is the most common localisation reported for equine meniscal injuries. Immunohistochemical stainings were performed using antibodies against high molecular weight neurofilaments and glial fibrillary acidic proteins. From a purely fundamental point of view, our work highlights for the first time the presence of Ruffini, Pacini and Golgi corpuscles in equine meniscus. They were found, isolated or in clusters and always located at the vicinity of blood vessels, at the level of the anterior horn of the equine medial meniscus. This morphological approach could serve as a basis for clinical studies, to evaluate the impact of these corpuscles on the poor sportive prognosis in equine meniscal tears.  相似文献   

17.
OBJECTIVE: To measure the ocular surface area, scleral thickness, and the aqueous and vitreous humor volumes in the canine and equine eye. Animals studied Fourteen canine and 16 equine cadaver eyes. PROCEDURE: Eyes were measured either fresh or following fixation in formalin. Ocular surface area was calculated using the fluid displacement method. Globes were hemisected and aqueous and vitreous humor were collected and quantitated. Scleral thickness was measured by digital caliper, by image projection, and by ultrasonic biomicroscopy (UBM). RESULTS: Mean +/- standard deviation (SD) scleral surface areas in canine and equine eyes were 12.87 +/- 2.24 and 57.23 +/- 5.63 cm2, respectively. Mean +/- SD aqueous humor volume was 0.77 +/- 0.24 in the dog and 3.04 +/- 1.27 mL in the horse. Mean vitreous humor volume was 1.7 +/- 0.86 mL for the canine eye and 26.15 +/- 4.87 mL for the equine eye. In canine and equine eyes, the sclera was thinnest at the ocular equator: 0.34 +/- 0.13 mm (canine) and 0.53 +/- 0.1 (equine). There were no significant differences between the direct caliper and projected thickness measurements or differences between measurements in the superior, inferior, nasal, and temporal quadrants of the eye. Scleral thickness in fresh tissue was greater than in fixed tissue at most sites. CONCLUSIONS: The UBM measurement method appeared to be most consistent and reproducible when compared to direct measurement techniques. Formalin fixation of the eyes was associated with significantly thinner scleral tissue than with fresh ocular tissue.  相似文献   

18.
OBJECTIVE: To determine penetration of topically and orally administered voriconazole into ocular tissues and evaluate concentrations of the drug in blood and signs of toxicosis after topical application in horses. ANIMALS: 11 healthy adult horses. PROCEDURE: Each eye in 6 horses was treated with a single concentration (0.5%, 1.0%, or 3.0%) of a topically administered voriconazole solution every 4 hours for 7 doses. Anterior chamber paracentesis was performed and plasma samples were collected after application of the final dose. Voriconazole concentrations in aqueous humor (AH) and plasma were measured via high-performance liquid chromatography. Five horses received a single orally administered dose of voriconazole (4 mg/kg); anterior chamber paracentesis was performed, and voriconazole concentrations in AH were measured. RESULTS: Mean +/- SD voriconazole concentrations in AH after topical administration of 0.5%, 1.0%, and 3.0% solutions (n = 4 eyes for each concentration) were 1.43 +/- 0.37 microg/mL, 2.35 +/- 0.78 microg/mL, and 2.40 +/- 0.29 microg/mL, respectively. The 1.0% and 3.0% solutions resulted in significantly higher AH concentrations than the 0.5% solution, and only the 3.0% solution induced signs of ocular toxicosis. Voriconazole was detected in the plasma for 1 hour after the final topically administered dose of all solutions. Mean +/- SD voriconazole concentration in AH after a single orally administered dose was 0.86 +/- 0.22 microg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that voriconazole effectively penetrated the cornea in clinically normal eyes and reached detectable concentrations in the AH after topical administration. The drug also penetrated noninflamed equine eyes after oral administration. Low plasma concentrations of voriconazole were detected after topical administration.  相似文献   

19.
Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 79 horses: 28 horses with various nonpemphigus dermatologic diseases, 21 horses with various nondermatologic diseases, and 30 normal horses. Pemphigus-like antibodies were detected in 6 horses: 3 normal horses with titers of 1:40, 2 horses with dermatophilosis at titers of 1:10 and 1:80, and 1 horse with lymphosarcoma at a titer of 1:320. It was concluded that equine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluorescence testing for whole immunoglobulin, IgG, IgM, and IgA was performed on skin lesions from 2 horses with dermatophilosis. Diffuse intercellular deposition of whole immunoglobulin and IgG was found in both horses. It was concluded that equine dermatophilosis is a potential source of misinterpretation and misdiagnosis in direct immunofluorescence testing.  相似文献   

20.
OBJECTIVE: To determine the efficacy of laparoscopic surgical techniques for repair of rectal lacerations in horses. STUDY DESIGN: Experimental study. ANIMAL OR SAMPLE POPULATION: Thirty-two segments of equine bowel placed in an equine pelvitrainer, 8 equine cadavers, and 3 normal horses. METHODS: In experiment 1, 3 laparoscopic intestinal-repair techniques were evaluated in an equine pelvitrainer: suturing with needle holders, with an automatic suture device, and stapling with a hernia stapler. In experiment 2, descending colon lacerations were sutured laparoscopically using needle holders in a pelvitrainer and in equine cadavers. In experiment 3, iatrogenic rectal lacerations were sutured laparoscopically with needle holders in horses under general anesthesia. These horses were evaluated for 7 days' postoperatively by clinical examination and blood and peritoneal fluid analysis. The horses were euthanatized 7 days' postoperatively and necropsied. The repaired colonic segments were collected for determination of bursting pressures, degree of luminal narrowing, and microscopic examination of the suture line. RESULTS: For the 3 techniques, there was no significant difference in repair time or degree of luminal narrowing, but bursting pressure was higher in segments repaired by use of needle holders or with the suture device. Colonic and rectal lacerations were sutured successfully with laparoscopic needle holders. No major complications were recorded. At necropsy, adequate healing was confirmed by a high bursting strength (>140 mm Hg) and by histologic examination. CLINICAL RELEVANCE: A novel laparoscopic method can be used to repair iatrogenic rectal lacerations in horses. Whether this method may be used for clinical cases of rectal tears must be investigated.  相似文献   

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