Surface-associated Hsp60 chaperonin of Leptospira interrogans serovar Autumnalis N2 strain as an immunoreactive protein.

The present study has been formulated in order to detect an immunoreactive protein whose identification can play a major role in the early diagnosis of disease. The identified protein will be produced by recombinant methods and used for the recombinant protein based ELISA. A comparison was made between the developed method and the gold standard MAT test to evaluate the serodiagnosis potential of the protein. The protein profile, immunoblot and MALDI-TOF analysis was carried out to identify the immunoreactive protein. The immunoreactive protein identified was used to develop ELISA for the diagnosis of leptospirosis using patients' sera with various clinical manifestations. The immunoreactive protein was identified as Leptospira GroEL chaperonin of molecular weight 60 kDa. The theoretical/experimental molecular weights, pI were found to be 58.5/60 kDa and 5.41/6, respectively. The overall results of the recombinant GroEL-IgM ELISAs showed cumulative sensitivity, specificity, positive predictive value, and negative predictive values of 90.6%, 94.9%, 94.6%, and 91.0%, respectively. The performance of such ELISA appeared better than that of any other serological tests previously evaluated for the diagnosis of leptospirosis in India. Thus, a highly conserved and immunogenic outer exposed GroEL protein during infection clearly merits further use in the serodiagnosis of leptospirosis.

Early pathogenesis of equine Streptococcus equi infection (strangles).

REASONS FOR PERFORMING STUDY: Little is known about entry and subsequent multiplication of Streptococcus equi following exposure of a susceptible horse. This information would have value in design of intranasal vaccines and understanding of shedding and protective immune responses. OBJECTIVES: To determine entry points and sites of subsequent replication and dispersion of S. equi at different times after intranasal infection or commingling exposure. METHODS: Previously unexposed horses and ponies were subjected to euthanasia 1, 3, 20 or 48 h following intranasal inoculation with biotin labelled or unlabelled S. equi CF32. Some ponies were inoculated with suspensions of equal numbers of CF32 and its mutants lacking capsule, S. equi M-like protein or streptolysin S. Others were infected by commingling exposure and subjected to euthanasia after onset of fever. Tonsils and lymph nodes were cultured for S. equi and tissues sectioned for histopathological examination and fluorescent microscopy. RESULTS: Tonsillar tissues of both the oro- and nasopharynx served as portals of entry. Entry was unexpectedly rapid but involved few bacteria. Small numbers of organisms were detected in tonsillar crypts, in adjacent subepithelial follicular tissue and draining lymph nodes 3 h after inoculation. By 48 h, clumps of S. equi were visible in the lamina propria. At onset of fever, tonsillar tissues and one or more mandibular and retropharyngeal lymph nodes were heavily infiltrated by neutrophils and long chains of extracellular S. equi. Mutant S. equi lacking virulence factors were not seen in draining lymph nodes. CONCLUSIONS: Although very small numbers of S. equi entered the lingual and nasopharyngeal tonsils, carriage to regional lymph nodes occurred within hours of inoculation. This observation, together with visual evidence of intracellular and extracellular multiplication of S. equi in tonsillar lymphoid tissue and lymph nodes over the following days, indicates involvement of potent antiphagocytic activity and failure of innate immune defences. RELEVANCE: Future research should logically address the tonsillar immune mechanisms involved including identification of effector cell(s) and antigens.

Streptococcus equi Infections in Horses: Guidelines for Treatment, Control, and Prevention of Strangles-Revised Consensus Statement.

This consensus statement update reflects our current published knowledge and opinion about clinical signs, pathogenesis, epidemiology, treatment, complications, and control of strangles. This updated statement emphasizes varying presentations in the context of existing underlying immunity and carrier states of strangles in the transmission of disease. The statement redefines the "gold standard" for detection of possible infection and reviews the new technologies available in polymerase chain reaction diagnosis and serology and their use in outbreak control and prevention. We reiterate the importance of judicious use of antibiotics in horses with strangles. This updated consensus statement reviews current vaccine technology and the importance of linking vaccination with currently advocated disease control and prevention programs to facilitate the eradication of endemic infections while safely maintaining herd immunity. Differentiation between immune responses to primary and repeated exposure of subclinically infected animals and responses induced by vaccination is also addressed.

Biotypes and ScM types of isolates of Streptococcus canis from diseased and healthy cats.

Lancefield group G Streptococcus canis is a component of the normal urogenital and pharyngeal flora of the cat. It is also frequently implicated in epizootics of severe disease in closed cat colonies and animal shelters. Given the importance of S canis as a feline pathogen and relative lack of published information on characteristics potentially associated with virulence, the authors have compared isolates from healthy and diseased cats in New York and California using fermentation profiles (biotype) and ScM sequences. With few exceptions, isolates associated with disease were biotype 1. Four alleles of scm were identified of which type 1 dominated in diseased cats. Type 4 allelic variants were found only in healthy cats and all but one were biotype 2. Type 2 and 3 alleles showed extensive N-terminal variation suggesting a plasminogen-binding site as found on the type 1 allele was absent. Cat antisera to ScM were opsonobactericidal, and these potentially protective antibodies increased during convalescence.

Detection of equine herpesvirus-specific effector and memory cytotoxic immunity in the equine upper respiratory tract.

Immunological protection of horses from equine herpesvirus-1 (EHV-1) infection and disease depends on the cooperation of virus-specific humoral and cellular immune responses. EHV-specific mucosal immunity may be an important component of such immune responses. This study demonstrates the induction of anti-EHV cytotoxic cellular immune responses in various mucosal and systemic lymphoid tissues associated with the upper respiratory tract (URT) of the horse. Four young horses (1-2 years of age) were inoculated intranasally with the Army 183 strain of EHV-1 and euthanized 1 week later. One untreated foal served as a non-infected control. Mucosa-associated tonsillar tissues, draining lymph nodes and PBMC were harvested. Virus-specific memory and effector cytolytic activity were individually assessed using 4 h chromium release assays, with and without in vitro restimulation with EHV-1, respectively. EHV-specific cytotoxic activity was detected ex vivo in several URT-associated mucosal lymphoid tissues of horses, particularly within the lining of the nasopharynx, a principal site of EHV-1 replication. This activity was also detected in the circulation of some horses 1 week post-challenge. Virus-specific memory cytotoxic activity was elevated in the circulation, and detectable in the draining lymph nodes of all horses following challenge infection.

Preliminary evaluation of a dual antigen ELISA to distinguish vaccinated from Leptospira infected horses.

Immunogenic proteins of Leptospira interrogans serovar Pomona type kennewicki (Lk) including Sph1, LigA, Hsp15 and LipL45 (Qlp42) are up-regulated in infected horses but are undetectable or expressed in trace amounts on cultured organisms. In contrast, LipL32 is abundant on cultured Lk and elicits infection antibody responses. The aim of this study was to develop an ELISA based on LipL32 or Lk sonicate and host-induced proteins to differentiate vaccine from infection serum antibody. IgG specific for recombinant Sph1, LigA, Lk90 (LigA; 379-1225 a.a), Hsp15, LipL45 and LipL32 of Lk were assayed in sera of horses infected naturally with Lk and before and after immunisation with serovar Pomona bacterin. Infection but not vaccine sera reacted strongly with Sph1, LigA and Lk90. LipL45 and Hsp15 reacted moderately with infection sera and weakly with vaccine sera. Lk sonicate and LipL32 reacted strongly with both infection and vaccine sera. As expected, culture-based vaccine failed to stimulate antibody to host-induced proteins. Therefore a dual antigen ELISA based on Lk sonicate or LipL32 combined with host-induced Sph1 and Lk90 will be valuable in differentiating infection from vaccine responses.

Multiple specificities of immunoglobulin M in equine fetuses infected with Leptospira interrogans indicate a competent immune response.

REASONS FOR PERFORMING STUDY: Foals of mares infected with Leptospira interrogans serovar Pomona type kennewicki (Lk) may be aborted/stillborn or delivered as healthy foals. Is fetal survival explained in part by the immune response of the fetus to Leptospira antigens? OBJECTIVES: To describe an outbreak of Leptospira abortion in which infected mares delivered dead/sick or normal foals and determine specificities of antibody in a collection of 54 fetuses from similar outbreaks. STUDY DESIGN: Outbreak investigation in combination with a case-control study of a larger set of samples from aborted fetuses. METHODS: Serology and polymerase chain reaction (PCR) on urine and amniotic fluids were used to diagnose infection during an outbreak of Leptospira abortion. Specificities of immunoglobulin (Ig)M, IgGa and IgGb for recombinant proteins of Lk in archived fluids of fetuses from similar outbreaks were compared by ELISA with those of fluids of fetuses not infected with Leptospira spp. RESULTS: Five fetuses of 11 infected mares in an outbreak survived in utero in the presence of persistent placental infection and were healthy at foaling. Fetuses of 6 mares in the outbreak were aborted or died soon after birth. Significantly greater (P<0.05) IgM reactivity with all recombinant proteins and with Lk sonicate was observed in 54 archived fluids from Leptospira infected fetuses than in fluids of 30 of non-Leptospira infected fetuses. Low levels of IgGa and IgGb specific for LipL32 and Lk sonicate and traces of LigA and Hsp15 specific IgGa were detected in a minority of archived fluids from Leptospira infected fetuses. CONCLUSION: Although mainly mediated by IgM, a high level of immune competence in aborted fetuses was evidenced by the multiplicity of Leptospira proteins targeted. This is likely to contribute to survival of foals in mares with evidence of placental infection at foaling as detailed in a typical outbreak.

Capsular hyaluronic acid of equine isolates of Streptococcus zooepidemicus is upregulated at temperatures below 35°C.

REASONS FOR PERFORMING STUDY: Streptococcus zooepidemicus causes opportunist respiratory and other infections in the horse. Capsule expression is highly variable and known to affect resistance to phagocytosis. Most clinical isolates producing small, dry colonies at 37°C produce mucoid colonies at temperatures below 35°C. OBJECTIVES: The aim was to understand the molecular basis of increased capsule expression by equine isolates of S. zooepidemicus at temperatures lower than 35°C. STUDY DESIGN: Cross-sectional observational study. METHODS: Capsule production by groups of equine S. zooepidemicus strains was determined at 23, 30, 35 and 37°C. Hyaluronidase (HylC) at 23 and 37°C was measured by quantitative enzyme-linked immunosorbent assay. Expressions of hasA and hylC at 23 and 37°C were measured by quantitative reverse transcriptase-PCR. The covRS genes in representative S. zooepidemicus were sequenced and checked for mutations. RESULTS: Colonies of randomly selected S. zooepidemicus strains became mucoid or showed marked increase in colony mucoidy following the change in temperature to 23°C. Expression of hasA at 23°C was 45- to 700-fold greater than at 37°C. Transcription of hylC at 23°C was 2.5- to 200-fold greater than at 37°C, yet enzyme concentrations in cultures were significantly higher at 37°C (P<0.05), suggesting that production of HylC is regulated post transcriptionally. The covRS gene in S. zooepidemicus was not mutated as seen in isolates of Streptococcus pyogenes with increased capsule production at 25°C. CONCLUSIONS: Sensitivity of capsule expression to temperature above 35°C but not HylC by the general population of equine S. zooepidemicus indicates that capsule is not required for extended colonisation nor for opportunist invasion. Instead, capsule production at lower than body temperature may reflect adaptation to life on skin and mucosal surfaces, where hyaluronic acid contributes to adhesion and resistance to desiccation. Pathogenicity of S. zooepidemicus following opportunist invasion is possibly dependent on factors other than capsule that may be co-regulated with HylC. The Summary is available in Chinese - see Supporting information.

Magnetic resonance imaging of experimental pyelonephritis in rabbits.

RATIONALE AND OBJECTIVES: An experimental model of acute focal pyelonephritis was developed in the New Zealand White rabbit, with initial imaging evaluation performed using high-field contrast-enhanced magnetic resonance (MR) in six animals. METHODS: The left kidney was visualized fluoroscopically after intravenous injection of iodinated contrast. A 0.1 mL mixture of agarose/Streptococcus faecalis was then injected percutaneously into the kidney at the corticomedullary junction with a 25-gauge needle. The animals were imaged at 1.5 tesla on day 5 after injection of bacteria. Breathhold T2-weighted and T1-weighted scans were acquired prior to contrast injection. Using an MR-compatible power injector, 0.3 mmol/kg gadoteridol was then administered, with both dynamic and delayed postcontrast scans obtained. The lesion was confirmed in each animal by gross and microscopic exam. RESULTS: The area of acute focal pyelonephritis was somewhat difficult to identify on precontrast scans. The lesion was mildly hyperintense on T2-weighted scans and slightly hypointense to isointense on T1-weighted scans relative to normal surrounding renal parenchyma. On early dynamic turbo-fast low-angle shot scans, the lesion could be identified due to differential tissue enhancement. Direct visualization of the lesion, with increased conspicuity relative to precontrast scans, was possible because of delayed positive enhancement at 60 seconds postcontrast. Lesion conspicuity, assessed by signal difference/noise ratio, increased from 0 +/- 5 precontrast to a peak of 12 +/- 6 at 60 seconds postcontrast (P = 0.003; n = 6). CONCLUSIONS: On dynamic contrast-enhanced MR, acute focal pyelonephritis can demonstrate transient positive contrast enhancement (using a contrast dose of 0.3 mmol/kg) relative to surrounding normal renal parenchyma. This appearance is due to hyperconcentration of the contrast agent in normal parenchyma and T2 effects. The pattern is opposite that seen in spiral computed tomography.

Magnetic resonance imaging in a spinal abscess model. Preliminary report.

RATIONAL AND OBJECTIVES: Magnetic resonance (MR) scan technique and lesion detectability were evaluated using a newly developed spinal abscess model in the New Zealand White rabbit. METHODS: To create the lesion, an epidural needle was inserted under fluoroscopic guidance in the lumbar region and advanced to penetrate the ligamentum flavum. Next, polyethylene tubing was fed through the needle into the epidural space. A mixed suspension of Staphylococcus aureus (Cowan I) and blue polystyrene microspheres then was injected. Lesions were evaluated by MR imaging in four animals at multiple time points (3, 6, and 9 days). Imaging was performed at 1.5 tesla using a surface coil. Precontrast T2-and T1-weighted scans were first obtained. The T1-weighted scans were acquired both with and without fat saturation, and were repeated after intravenous contrast administration. The contrast agent used was gadoteridol (gadolinium HP-DO3A or ProHance) at a dose of 0.3 mmol/kg. RESULTS: On prospective film review, postcontrast scans proved superior for lesion detection. A spinal abscess could be identified postcontrast in all cases, irrespective of the use of fat saturation. The next best imaging technique for lesion detection was the T2-weighted scan, with 5 of 8 lesions noted thereon. Visualization of lesion margins proved to be a primary factor in prospective lesion identification. Region of interest image analysis demonstrated the postcontrast scans to be superior to all precontrast scan techniques for conspicuity of the interface between the abscess and the compressed spinal cord, with these results statistically significant. The lesions were characterized histologically by infiltrates of heterophils into the meninges and outer spinal cord with accompanying mild hemorrhage, fibrin exudation, and bacterial colonies. The lesions in three animals were confirmed to be in the epidural space, with the lesion in one animal in the subdural space. CONCLUSIONS: The current animal model was developed to study spine infection and, specifically, imaging characteristics and lesion detectability on MR. With the increased use of epidural catheters for pain management and the large number of acquired immunodeficiency syndrome cases, epidural infection is becoming an increasingly important clinical problem. Imaging technique, in particular the use of intravenous contrast, is critical for lesion detection and evaluation.

The use of gadolinium-BOPTA on magnetic resonance imaging in brain infection.

RATIONALE AND OBJECTIVES: The use of gadolinium (Gd)-BOPTA as a magnetic resonance contrast agent for central nervous system disease was studied in a canine brain abscess model. METHODS: A Streptococcus faecalis brain abscess was evaluated in five dogs at 1.5T. Imaging was performed during the late cerebritis stage, at 5 to 7 days after surgery. Magnetic resonance scans were acquired before and at 1, 5, 15, 30, 45, and 60 minutes after contrast administration, using a dose of 0.1 mmol/kg. Scans also were acquired both before and after contrast injection with the implementation of magnetization transfer. RESULTS: Lesion enhancement, quantified by region-of-interest measurement, peaked at 5 minutes after contrast injection. Both the increase in lesion enhancement from 1 to 5 minutes after injection and the decrease from 5 to 15 minutes after injection, although small, were statistically significant (P < 0.004 and P < 0.03, respectively). The application of magnetization transfer improved lesion enhancement, as measured by signal difference/noise, by 39%. This result also was statistically significant (P < 0.001). CONCLUSIONS: In intraparenchymal brain infection, Gd-BOPTA provides effective lesion enhancement when used at a dose of 0.1 mmol/kg. Further research is needed to compare the magnitude of enhancement achieved with Gd-BOPTA, which has weak protein binding and both hepatobiliary and renal excretion, with that with Gd chelates, which have pure renal excretion.

Detectability of early brain meningitis with magnetic resonance imaging.

RATIONALE AND OBJECTIVES: The ability of high-field (1.5 T) magnetic resonance imaging (MRI) to detect early brain meningitis was evaluated in a canine model. Contrast dose, timing postinjection, and imaging technique (specifically the use of magnetization transfer) were assessed. METHODS: Imaging of five canines was performed at 1.5 T 24 hours after injection of Cowans staphylococcus into the cisterna magna. Two control animals also were imaged using the same protocol, with one animal receiving a cisternal injection of nutrient broth only and the other no injection. Contrast doses of 0.1, 0.3, and 0.8 mmol/kg gadoteridol (Gd HP-DO3A or Pro-Hance) were compared. Scans were performed at 2, 12, and 22 minutes after an initial injection of 0.1 mmol/kg. At each time point, paired T1-weighted scans with and without magnetization transfer (MT) were acquired. Thirty minutes after the initial injection of contrast, a supplemental dose of 0.2 mmol/kg was given (for a cumulative dose of 0.3 mmol/kg). Scans were then repeated at 2, 12, and 22 minutes after this dose was administered. A second supplemental contrast injection of 0.5 mmol/kg (for a cumulative dose of 0.8 mmol/kg) was given at 70 minutes, and immediate postinjection scans with and without MT were acquired. RESULTS: In the animals receiving a cisternal injection of bacteria, the degree of meningeal enhancement was greatest at 0.8 mmol/kg, intermediate at 0.3 mmol/kg, and least at 0.1 mmol/kg. These conclusions were constant whether imaging was performed with or without MT. Scans in control studies did not demonstrate abnormal meningeal enhancement. High-contrast dose, MT, and acquisition of immediate postcontrast scans all resulted in statistically significant improvement. On masked film review, abnormal meningeal enhancement was noted in only 2 of 5 experimental dogs at a dose of 0.1 mmol/kg (regardless of the use of MT) compared with all animals at a dose of 0.3 mmol/kg. In 18 of 37 dogs (paired scans with and without MT), when abnormal enhancement was noted, the use of MT improved the visualization of abnormal meningeal enhancement. CONCLUSIONS: In early brain meningitis, high-contrast dose (0.3 mmol/kg), MT, and scanning immediately after injection improve detection of abnormal meningeal enhancement, thus facilitating the diagnosis of meningitis. Of these factors, contrast dose is the most important.

Biochemical differences among human and animal streptococci of Lancefield group C or group G.

Pyogenic streptococci of Lancefield group C or group G from human or animal sources were examined with a view to increasing the number of diagnostic tests useful for their differentiation. Human strains of group G produced L-prolyl-L-arginine aminopeptidase but isolates of Streptococcus equisimilis (group C) did not. Tests for alpha-L-glutamate aminopeptidase together with fermentation of glycogen or sorbitol distinguished S. dysgalactiae from strains of S. equisimilis isolated from animals. It was confirmed that fermentation tests were helpful in the study of S. equi and S. zooepidemicus and that enzyme reactions helped distinguish between S. canis and the human strains of group G.

Properties of a protective protein antigen of Erysipelothrix rhusiopathiae.

Erysipelothrix rhusiopathiae is a widely distributed mucosal commensal of the alimentary tracts of vertebrates. Antibodies to a 66-64 kDa protein released from the cell surface have been shown to be involved in protective immunity. Mice immunized with the purified 66-64 kDa protein from strain T28, serotype 2b were protected against challenge by the United States challenge strain E1-6P (serotype 1a) and by the official German challenge strain Frankfurt 1 (serotype N). Thus, protection is not serotype specific, a result consistent with previous observations that polysaccharide, non-proteinaceous antigens are the type specific antigens useful in serotyping. The 66-64 kDa protein appears to be most immunogenic when complexed to glycolipid. This may be due to an adjuvant effect of polysaccharide antigens. Further studies on the correlation between antibody titers to the 66-64 kDa protein and protection in pigs, turkeys and mice in vivo should be helpful in developing a basis for an in vitro assay to replace the mouse protection test in vaccine testing.

The protective M proteins of the equine group C streptococci.

The group C streptococci are the most commonly isolated bacteria from disease states in the horse. Important virulence factors of S. equi and S. zooepidemicus are the hyaluronic acid capsule and the antiphagocytic fibrillar M protein located on the surface of the cell wall and extending into and through the capsule. The hyaluronic acid capsule is non-antigenic and so is not involved in protective immunity. The M protein, a superantigen, elicits very strong B and T cell responses that may result in protective immunity mediated by opsonic antibodies in plasma and by locally synthesized IgG and IgA on the pharyngeal mucosa. However, vaccines based on acid or mutanolysin extracted M protein do not confer a high level of protection against field exposure. Protective antibodies to S. equi or S. zooepidemicus can in part be assayed by the bactericidal test that measures opsonization for equine neutrophils. A mouse-challenge model has also been used to test immunizing potency of streptococcal extracts and in a passive protection test for protective antibody. There is as yet no means of distinguishing protective opsonic or mucosal antibodies from other antibodies produced against the many epitopes on the M molecule.

In vivo pathogenicity and resistance to phagocytosis of Streptococcus equi strains with different levels of capsule expression.

The glossy non-encapsulated strain of Steptococcus equi, NCTC 9682, was compared with the matt strain Hidaka/95/2 which expresses a medium sized capsule and with the mucoid CF32 which expresses a large sized capsule in phagocytosis assays and for virulence in inoculated horses. The three strains, NCTC 9682, Hidaka /95/2 and CF32 produced 2.0, 3.1, and 5.3 mg/g wet cells respectively after 3 h incubation, but similar amounts of M-like proteins, cytotoxin and mitogen. NCTC 9682 showed no resistance to phagocytosis by equine neutrophils regardless of the presence of opsonin while strains Hidaka /95/2 and CF32 showed almost complete resistance to phagocytosis. Furthermore, NCTC 9682 produced no clinical disease although it infected the guttural pouch and caused seroconversion. Typical strangles with guttural pouch invasion was observed in all horses infected with encapsulated strains.

Purification and characterization of equine complement factor C3.

A rapid method for purifying equine C3 which yields milligram quantities of pure C3 is described. Protein from equine plasma was selectively precipitated with polyethylene glycol, and the C3 was purified by anionic and cationic exchange HPLC. The yield from this procedure was 12%. The purified C3 was composed of an alpha chain (118 kD) and a beta chain (68 kD) linked by at least one disulfide bond, and it had an isoelectric point of 4.7. Amino acid analysis indicated a strong conservation of amino acid usage between equine and human C3. The N-terminal sequences of the alpha and beta chains were homologous to human, mouse, and rat C3, and activation of C3 produced breakdown products similar in molecular weight to C3b and iC3b of other species. Equine C3 appeared to be functionally dependent upon a reactive thiolester as treatment of fresh equine serum with methylamine abrogated its hemolytic activity.

Antibody isotypes in sera of equine fetuses aborted due to Leptospira interrogans serovar pomona-type kennewicki infection.

Leptospira-specific antibody isotypes in sera of late term equine fetuses aborted due to Leptospira interrogans serovar pomona-type kennewicki infection were characterized and compared with those of their dams. IgM was the dominant Leptospira-Specific isotype in both fetuses and mares. However, IgGa was the isotype in highest concentration in petal sera and strong Leptospira-specific IgGa but no IgGb and little or no IgG(T) were detected. In contrast, although IgGb was quantitatively the dominant isotype in mares serum, Leptospira-specific serum IgG in aborting mares was dominated by IgG(T) but also included large amounts of IgGa and IgGb. IgGa and IgGb were quantitatively the dominant isotypes in sera of fetuses and mares, respectively. Affinity purified IgGa from fetuses did not agglutinate leptospires but serum devoid of IgGa did, suggesting that IgM is the principal agglutinating antibody. It is concluded that the equine fetus is deficient in IgGb and IgG(T) synthesis.

Serum and mucosal antibody isotype responses to M-like protein (SeM) of Streptococcus equi in convalescent and vaccinated horses.

Equine strangles, caused by the clonal pathogen Streptococcus equi, is a source of serious economic loss despite the widespread use of commercial vaccines. The anti-phagocytic 58 kDa M-like protein (SeM) is an important protective antigen. The objective of this study was to define differences, if any, between SeM-specific convalescent serum and mucosal IgA and IgG subisotypes and those induced by vaccination with commercial strangles vaccine. SeM-specific opsonophagocytic IgGb was the predominant serum antibody in horses intramuscularly vaccinated or recently recovered from infection. Infection also induced high levels of specific opsonophagocytic serum IgGa during and shortly after S. equi infection whereas vaccination stimulated only low levels of serum IgGa. Specific serum IgGc and opsonophagocytic IgA were present at very low levels following infection or vaccination. A strong specific mucosal antibody response occurred during the acute and convalescent phases of infection whereas vaccinated horses made no mucosal response. Specific IgGb was generally predominant in nasopharyngeal washings during the acute phase but was replaced by specific IgA during convalescence. SeM-specific mucosal IgGa and IgG(T) but not IgGc were detected only during the acute and early convalescent phase. The results therefore indicate that vaccination, although inducing SeM-specific serum isotype responses qualitatively and quantitatively similar to those seen in convalescence, did not induce mucosal responses. This suggests that mucosal immunity may be important in acquired resistance to strangles.

Equine glomerulonephritis and renal failure associated with complexes of group-C streptococcal antigen and IgG antibody.

A 12-year-old thoroughbred gelding died from diffuse global glomerulonephritis, 3 months after a lower respiratory infection from which Streptococcus zooepidemicus was isolated. Immunopathological studies (immunofluorescence, immunodiffusion, immunoperoxidase testing and immunoblotting) indicated the presence of an immune reactant renal disease associated with IgG antibody and streptococcal antigens.