Association of the neutrophil-lymphocyte ratio with outcome in sick hospitalized neonatal foals.

BACKGROUND: The neutrophil-lymphocyte ratio (NLR) in human medicine is an objective biomarker that reflects prognosis. The NLR as an independent biomarker to help predict nonsurvival in hospitalized neonatal foals has not been thoroughly interrogated. OBJECTIVES/HYPOTHESIS: Retrospectively evaluate if the NLR at admission is associated with nonsurvival in sick hospitalized foals <4 days old. We hypothesized that a lower NLR will be associated with nonsurvival. ANIMALS: One thousand one hundred ninety-six client-owned foals <4 days old of any breed and sex: 993 hospitalized foals and 203 healthy foals. METHODS: Retrospective multicenter study. Medical records of foals presenting to 3 equine referral hospitals were reviewed. Foals were included if they had complete CBCs, sepsis scores, and outcome data. The NLR was calculated by dividing the absolute neutrophil count by the absolute lymphocyte count. Data were analyzed by nonparametric methods and univariate analysis. RESULTS: Of the 993 sick hospitalized foals, 686 were sick nonseptic and 307 were septic. The median NLR was lower in sick hospitalized foals (median [95% confidence interval], 3.55 [0.5-13.9]) compared with healthy foals (6.61 [3.06-18.1]). Septic foals had the lowest NLR (2.00 [0.20-9.71]). The NLR was lower in nonsurviving (1.97 [1.67-2.45]) compared with surviving foals (4.10 [3.76-4.33]). Nonsurviving septic foals had the lowest NLR (1.47 [1.70-3.01]). Foals with a NLR of <3.06 or <1.6 at admission had odds ratio of 3.21 (2.24-4.29) and 4.03 (2.86-5.67) for nonsurvival, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: A NLR < 3.06 at admission in sick hospitalized foals is readily available and clinically useful variable to provide prognostic information.

Serum and cerebrospinal fluid phosphorylated neurofilament heavy protein concentrations in equine neurodegenerative diseases.

BACKGROUND: Currently, there is little information regarding the concentrations of phosphorylated neurofilament heavy protein (pNfH) in the serum and cerebrospinal fluid (CSF) of horses with neurodegenerative diseases. Specifically, pNfH concentrations have not yet been evaluated in horses with equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy (eNAD/EDM). OBJECTIVES: To determine pNfH concentrations using a commercial enzyme-linked immunosorbent assay (ELISA) in serum and CSF from control horses and horses with eNAD/EDM, cervical vertebral compressive myelopathy (CVCM) and Shivers. STUDY DESIGN: Case-control study using biobanked samples from diseased horses and prospective or biobanked samples from control horses. METHODS: The pNfH ELISA was performed on samples from horses diagnosed with eNAD/EDM (n = 64), CVCM (n = 26) and Shivers (n = 9) and 51 neurologically normal control horses. RESULTS: Median and 95% confidence interval (CI) serum pNfH concentrations in control, CVCM, and eNAD/EDM horses were 0.08 ng/mL (0.07-0.15), 0.07 ng/mL (0.07-0.15) and 0.07 ng/mL (0.07-1.13), respectively. Serum pNfH concentrations were below the limit of detection (<0.07 ng/mL) for all Shivers horses. CSF pNfH concentrations in control, CVCM-, eNAD/EDM- and Shivers-affected horses were 1.26 ng/mL (1.06-1.5), 3.07 ng/mL (1.15-29.9), 1.78 ng/mL (1.5-2.28) and 1.39 ng/mL (0.74-3.89), respectively. CSF pNfH concentrations were significantly higher in CVCM (P = .001) and eNAD/EDM (P  = .01) affected horses compared to control horses. Serum pNfH concentrations >1 ng/mL were significantly associated with eNAD/EDM (P = .01) with only 12% sensitivity but 99% specificity. CSF pNfH concentrations >3 ng/mL were significantly associated with CVCM (P = .0002), with 50% sensitivity and 86% specificity. MAIN LIMITATIONS: A limited number of control horses tested were <1 year of age. CONCLUSIONS: Serum pNfH concentrations are specifically increased (>1 ng/mL) in some horses with eNAD/EDM. Increased CSF pNfH concentrations (>3 ng/mL) can be observed with eNAD/EDM or CVCM.

Horses affected by EPM have increased sCD14 compared to healthy horses.

Equine protozoal myeloencephalitis (EPM) is a debilitating neurologic disease affecting horses across the Americas. Gaps in understanding the inflammatory immune response in EPM-affected horses create difficulties with diagnosis and treatment, subsequently negatively impacting the prognosis of affected horses. The purpose of the current study was to evaluate circulating levels of the inflammatory immune marker soluble CD14 (sCD14), in horses with EPM (n = 7) and determine if they differed from healthy neurologically normal horses (n = 6). Paired sera and cerebrospinal fluid (CSF) samples were analyzed for sCD14. Inclusion criteria for EPM horses consisted of the presence of neurologic signs consistent with EPM, Sarcocystis neurona surface antigens 2, 4/3 (SnSAG 2, 4/3) ELISA serum: CSF antibody ratio ≤ 100, and a postmortem diagnosis of EPM. Control horses were neurologically normal, healthy horses with SnSAG 2, 4/3 ELISA serum: CSF antibody ratios of > 100. Serum anti-Sarcocystis neurona antibodies indicate that healthy control horses were exposed to S. neurona but resistant to developing clinical EPM. EPM cases had significantly greater concentrations of sCD14 in CSF samples compared to control horses and increased serum sCD14 concentrations. A positive correlation between sCD14 serum and CSF concentrations was observed in EPM-affected horses but not healthy horses. Soluble CD14 is an inflammatory marker, and the study results suggest it is elevated in EPM patients. When performed in conjunction with clinical evaluation and standard antibody testing, there may be potential for sCD14 to be utilized as a correlate for EPM.

Comparative H-gal-GP and H11 specific antibody binding in equine cyathostomins.

The biological-based vaccine (Barbervax®) generates effective antibodies against the biologically essential H-gal-GP and H11 protein complex of the ruminant parasite Haemonchus contortus to target and kill the parasites after taking a blood meal. A comparative analysis of several parasite genera was performed to determine if a similar protein complex or one that is recognized by H-gal-GP and H11 specific antibodies was present. If so, it suggests the vaccine could be effective for other nematode parasites. Ancylostoma caninum, H. contortus, equine cyathostomins, bovine Bunostomum phlebotomum, Dracunculus lutrae, Parascaris sp., Ixodes scapularis, Amblyomma americanum, Dirofilaria immitis and Brugia malayi were evaluated for specific antibody binding using hyperimmunized antibodies against H-gal-GP and H11 native proteins. Of the parasites evaluated, specific and reproducible staining was observed in H. contortus and adult and encysted cyathostomins only. To further evaluate the similar reactivities between cyathostomins and H. contortus, cross-reactivity of equine serum with antibodies to cyathostomins on a H. contortus adult histology cross-section was observed using immunofluorescence. These findings pave the way for future studies on the safety and efficacy of H-gal-GP and H11 protein complex as a potential control for cyathostomins.

Small sarcocysts can be a feature of experimental infections with Sarcocystis neurona merozoites.

Several reports indicate the presence of small tissue cysts associated with Sarcocystis neurona infections. Several failed attempts to develop tissue cysts in potential intermediate host using in vitro derived parasites originally isolated from horses with equine protozoal myeloencephalitis suggest that the experimental methods to achieve bradyzoites with those isolates was not possible. Those prior studies reported the lack of detectable sarcocysts based on histology and in vivo feeding trials. A recent report of successful production and detection of small sarcocysts triggered us to review archived tissues from earlier experimental infection studies. The retrospective review sought to determine if small sized sarcocysts were not detected due to their relatively smaller size and infrequency as compared to larger sized sarcocysts produced with other isolates in these experimental inoculation trials. Tissues from two prior in vivo inoculation studies, involving in vitro-produced parasites inoculated into laboratory-reared cats and raccoons, were re-examined by immunohistochemistry staining to more easily detect the tissue cysts. In the experimental cat study no small tissue cysts were seen, consistent with the original publication results. However, in the experimental raccoon study, one raccoon inoculated with an EPM-derived isolate, SN-UCD1, had small sarcocysts not reported in the original publication. This retrospective study suggests that much closer scrutiny of tissues, including the use of immunohistochemistry on tissue sections is required to detect the smaller S. neurona sarcocysts associated with the experimental inoculations of the isolates originally derived from horses with EPM.

Testing the Sarcocystis neurona vaccine using an equine protozoal myeloencephalitis challenge model.

Equine protozoal myeloencephalitis (EPM) is an important equine neurologic disorder, and treatments for the disease are often unrewarding. Prevention of the disease is the most important aspect for EPM, and a killed vaccine was previously developed for just that purpose. Evaluation of the vaccine had been hampered by lack of post vaccination challenge. The purpose of this study was to determine if the vaccine could prevent development of clinical signs after challenge with Sarcocystis neurona sporocysts in an equine challenge model. Seventy horses that were negative for antibodies to S. neurona and were neurologically normal were randomly assigned to vaccine or placebo groups and divided into short-term duration of immunity (study #1) and long-term duration of immunity (study #2) studies. S. neurona sporocysts used for the challenge were generated in the opossum/raccoon cycle isolate SN 37-R. Study #1 horses received an initial vaccination and a booster, and were challenged 34days post second vaccination. Study #2 horses received a vaccination and two boosters and were challenged 139days post third vaccination. All horses in study #1 developed neurologic signs (n=30) and there was no difference between the vaccinates and controls (P=0.7683). All but four horses in study #2 developed detectable neurologic deficits. The neurologic signs, although not statistically significant, were worse in the vaccinated horses (P=0.1559). In these two studies, vaccination with the S. neurona vaccine failed to prevent development of clinical neurologic deficits.

Sarcocystis neurona manipulation using culture-derived merozoites for bradyzoite and sporocyst production.

Equine protozoal myeloencephalitis (EPM) remains a significant central nervous system disease of horses in the American continents. Sarcocystis neurona is considered the primary causative agent and its intermediate life stages are carried by a wide host-range including raccoons (Procyon lotor) in North America. S. neurona sarcocysts mature in raccoon skeletal muscle and can produce central nervous system disease in raccoons, mirroring the clinical presentation in horses. The study aimed to develop laboratory tools whereby the life cycle and various life stages of S. neurona could be better studied and manipulated using in vitro and in vivo systems and compare the biology of two independent isolates. This study utilized culture-derived parasites from S. neurona strains derived from a raccoon or from a horse to initiate raccoon infections. Raccoon tissues, including fresh and cryopreserved tissues, were used to establish opossum (Didelphis virginiana) infections, which then shed sporocyts with retained biological activity to cause encephalitis in mice. These results demonstrate that sarcocysts can be generated using in vitro-derived S. neurona merozoites, including an isolate originally derived from a naturally infected horse with clinical EPM. This study indicates the life cycle can be significantly manipulated in the laboratory without affecting subsequent stage development, allowing further purification of strains and artificial maintenance of the life cycle.

Vitamin D Metabolites and Their Association with Calcium, Phosphorus, and PTH Concentrations, Severity of Illness, and Mortality in Hospitalized Equine Neonates.

BACKGROUND: Hypocalcemia is a frequent abnormality that has been associated with disease severity and outcome in hospitalized foals. However, the pathogenesis of equine neonatal hypocalcemia is poorly understood. Hypovitaminosis D in critically ill people has been linked to hypocalcemia and mortality; however, information on vitamin D metabolites and their association with clinical findings and outcome in critically ill foals is lacking. The goal of this study was to determine the prevalence of vitamin D deficiency (hypovitaminosis D) and its association with serum calcium, phosphorus, and parathyroid hormone (PTH) concentrations, disease severity, and mortality in hospitalized newborn foals. METHODS AND RESULTS: One hundred newborn foals ≤72 hours old divided into hospitalized (n = 83; 59 septic, 24 sick non-septic [SNS]) and healthy (n = 17) groups were included. Blood samples were collected on admission to measure serum 25-hydroxyvitamin D3 [25(OH)D3], 1,25-dihydroxyvitamin D3 [1,25(OH) 2D3], and PTH concentrations. Data were analyzed by nonparametric methods and univariate logistic regression. The prevalence of hypovitaminosis D [defined as 25(OH)D3 <9.51 ng/mL] was 63% for hospitalized, 64% for septic, and 63% for SNS foals. Serum 25(OH)D3 and 1,25(OH) 2D3 concentrations were significantly lower in septic and SNS compared to healthy foals (P<0.0001; P = 0.037). Septic foals had significantly lower calcium and higher phosphorus and PTH concentrations than healthy and SNS foals (P<0.05). In hospitalized and septic foals, low 1,25(OH)2D3 concentrations were associated with increased PTH but not with calcium or phosphorus concentrations. Septic foals with 25(OH)D3 <9.51 ng/mL and 1,25(OH) 2D3 <7.09 pmol/L were more likely to die (OR=3.62; 95% CI = 1.1-12.40; OR = 5.41; 95% CI = 1.19-24.52, respectively). CONCLUSIONS: Low 25(OH)D3 and 1,25(OH)2D3 concentrations are associated with disease severity and mortality in hospitalized foals. Vitamin D deficiency may contribute to a pro-inflammatory state in equine perinatal diseases. Hypocalcemia and hyperphosphatemia together with decreased 1,25(OH)2D3 but increased PTH concentrations in septic foals indicates that PTH resistance may be associated with the development of these abnormalities.

A new trivalent SnSAG surface antigen chimera for efficient detection of antibodies against Sarcocystis neurona and diagnosis of equine protozoal myeloencephalitis.

Enzyme-linked immunosorbent assays (ELISAs) based on the SnSAG surface antigens of Sarcocystis neurona provide reliable detection of infection by the parasite. Moreover, accurate serodiagnosis of equine protozoal myeloencephalitis (EPM) is achieved with the SnSAG ELISAs by measuring antibodies in serum and cerebrospinal fluid (CSF) to reveal active infection in the central nervous system. Two independent ELISAs based on recombinant (r)SnSAG2 or a chimeric fusion of SnSAG3 and SnSAG4 (rSnSAG4/3) are currently used together for EPM serodiagnosis to overcome varied antibody responses in different horses. To achieve reliable antibody detection with a single ELISA instead of 2 separate ELISAs, rSnSAG2 was fused with rSnSAG4/3 into a single trivalent protein, designated rSnSAG2/4/3. Paired serum and CSF from 163 horses were tested with all 3 ELISAs. When the consensus antibody titers obtained with the rSnSAG2 and rSnSAG4/3 ELISAs were compared to the single SAG2/4/3 ELISA titers, Spearman rank correlation coefficients of ρ = 0.74 and ρ = 0.90 were obtained for serum and CSF, respectively, indicating strong agreement between the tests. When the rSnSAG2 and rSnSAG4/3 consensus serum-to-CSF titer ratio was compared to the rSnSAG2/4/3 serum-to-CSF titer ratio, the Spearman correlation coefficient was ρ = 0.87, again signifying strong agreement. Importantly, comparing the diagnostic interpretation of the serum-to-CSF titer ratios yielded a Cohen kappa value of 0.77. These findings suggest that the single ELISA based on the trivalent rSnSAG2/4/3 will provide serologic and diagnostic results that are highly comparable to the consensus of the 2 independent ELISAs based on rSnSAG2 and rSnSAG4/3.

Equine protozoal myeloencephalitis.

Equine protozoal myeloencephalitis (EPM) can be caused by either of 2 related protozoan parasites, Sarcocystis neurona and Neospora hughesi, although S. neurona is the most frequent etiologic pathogen. Horses are commonly infected, but clinical disease occurs infrequently; the factors influencing disease occurrence are not well understood. Risk factors for the development of EPM include the presence of opossums and prior stressful health-related events. Attempts to reproduce EPM experimentally have reliably induced antibody responses in challenged horses but have not consistently produced acute neurologic disease. Diagnosis and options for treatment of EPM have improved over the past decade.

Factors predictive of abnormal results for computed tomography of the head in horses affected by neurologic disorders: 57 cases (2001-2007).

OBJECTIVE: To determine neurologic indications associated with abnormal results for computed tomography (CT) imaging of the head of horses affected by neurologic disorders. DESIGN: Retrospective case series. ANIMALS: 57 horses. PROCEDURES: Signalment, history, clinical abnormalities, and clinicopathologic findings were obtained from medical records of horses examined because of neurologic disorders, and precontrast and postcontrast CT images of the head were reviewed. Data were analyzed by use of univariate and multivariate logistic regression. RESULTS: For a horse with abnormal mentation, odds of having abnormal results for CT imaging of the head was 30 times (95% confidence interval [CI], 2.36 to 374.63) the odds for a similar horse without abnormal mentation. For a horse with cranial nerve deficits, odds of having abnormal results for CT imaging of the head was 11 times (95% CI, 1.00 to 127.96) the odds for a similar horse without cranial nerve deficits. For a horse with seizure-like activity, odds of having abnormal results for CT imaging of the head was 0.05 times (95% CI, 0 to 0.90) the odds for a similar horse without seizures. CONCLUSIONS AND CLINICAL RELEVANCE: These results suggested that alterations in consciousness and cranial nerve deficits were strong predictors of abnormal CT findings for the head of affected horses. Thus, CT can be a useful complementary diagnostic test in horses with these neurologic deficits. In contrast, alternative diagnostic tests (eg, electroencephalography and magnetic resonance imaging) should be considered in horses with seizure-like activity that do not have head trauma or cranial nerve deficits.

Use of kinetic gait analysis for detection, quantification, and differentiation of hind limb lameness and spinal ataxia in horses.

OBJECTIVE: To evaluate use of kinetic gait analysis for detection, quantification, and differentiation of hind limb lameness and spinal ataxia in horses. DESIGN: Prospective clinical study. ANIMALS: 36 horses. Procedures-Kinetic gait analysis with a force plate was performed for 12 clinically normal horses, 12 horses with hind limb lameness, and 12 horses with spinal ataxia. Kinetic variables were compared among groups, correlated to subjective grading, and used to build predictive models to assess the accuracy of discrimination. RESULTS: Subsets of kinetic variables were characteristically altered in ataxic and lame gaits. Ataxic horses had significantly increased lateral force peak and variation in vertical force peaks in both hind limbs. Lame horses had significantly decreased vertical force peak and increased variation in vertical force peaks only in the lame hind limb. These variables were used to differentiate between spinal ataxia and hind limb lameness with excellent accuracy. There were significant correlations between a subset of kinetic variables and subjective lameness and neurologic grades. CONCLUSIONS AND CLINICAL RELEVANCE: Kinetic gait variables, specifically lateral force peak and the variation in vertical force, can be used to support the differential diagnosis between spinal ataxia and hind limb lameness in horses. Kinetic gait analysis may also be applied for quantification of equine hind limb gait abnormalities as well as confirming lack of lameness and ataxia in soundness examinations.

A multicenter case-control study of risk factors for equine protozoal myeloencephalitis.

OBJECTIVE: To identify risk factors for equine protozoal myeloencephalitis (EPM) among horses examined at 11 equine referral hospitals. DESIGN: Case-control study. ANIMALS: 183 horses with EPM, 297 horses with neurologic disease other than EPM (neurologic controls), and 168 horses with non-neurologic diseases (non-neurologic controls) examined at 11 equine referral hospitals in the United States. PROCEDURES: A study data form was completed for all horses. Data were compared between the case group and each of the control groups by means of bivariate and multivariate polytomous logistic regression. RESULTS: Relative to neurologic control horses, case horses were more likely to be > or = 2 years old and to have a history of cats residing on the premises. Relative to non-neurologic control horses, case horses were more likely to be used for racing or Western performance. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that cats may play a role in the natural epidemiology of EPM, that the disease is less common among horses < 2 years of age relative to other neurologic diseases, and that horses used for particular types of competition may have an increased risk of developing EPM.

Pharmacokinetics of voriconazole following intravenous and oral administration and body fluid concentrations of voriconazole following repeated oral administration in horses.

OBJECTIVE: To determine the pharmacokinetics of voriconazole following IV and PO administration and assess the distribution of voriconazole into body fluids following repeated PO administration in horses. ANIMALS: 6 clinically normal adult horses. PROCEDURES: All horses received voriconazole (10 mg/kg) IV and PO (2-week interval between treatments). Plasma voriconazole concentrations were determined prior to and at intervals following administration. Subsequently, voriconazole was administered PO (3 mg/kg) twice daily for 10 days to all horses; plasma, synovial fluid, CSF, urine, and preocular tear film concentrations of voriconazole were then assessed. RESULTS: Mean +/- SD volume of distribution at steady state was 1,604.9 +/- 406.4 mL/kg. Systemic bioavailability of voriconazole following PO administration was 95 +/- 19%; the highest plasma concentration of 6.1 +/- 1.4 microg/mL was attained at 0.6 to 2.3 hours. Mean peak plasma concentration was 2.57 microg/mL, and mean trough plasma concentration was 1.32 microg/mL. Mean plasma, CSF, synovial fluid, urine, and preocular tear film concentrations of voriconazole after long-term PO administration were 5.163 +/- 1.594 microg/mL, 2.508 +/- 1.616 microg/mL, 3.073 +/- 2.093 microg/mL, 4.422 +/- 0.8095 microg/mL, and 3.376 +/- 1.297 microg/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that voriconazole distributed quickly and widely in the body; following a single IV dose, initial plasma concentrations were high with a steady and early decrease in plasma concentration. Absorption of voriconazole after PO administration was excellent, compared with absorption after IV administration. Voriconazole appears to be another option for the treatment of fungal infections in horses.

Outbreak of neurologic disease caused by equine herpesvirus-1 at a university equestrian center.

BACKGROUND: Equine herpesvirus type 1 (EHV-1) infection causes neurologic disease in horses. However, risk factors for the disease and long-term prognosis are poorly characterized. HYPOTHESIS: There are identifiable risk factors for equine herpes-1 myeloencephalopathy. ANIMALS: The entire population of 135 horses housed within the equestrian facility. METHODS: A descriptive study investigated the clinical, serologic, virologic, and management aspects of an outbreak of EHV-1 myeloencephalopathy. RESULTS: Out of 135 horses at the facility, 117 displayed signs of EHV-1 infection. Forty-six horses developed neurologic deficits characterized by symmetrical hind limb ataxia and weakness. Twelve horses that developed neurologic deficits became recumbent and did not survive. The development of severe neurologic deficits during the outbreak was associated with the presence of residual deficits at the 6-month examination. Within 1 year of the outbreak onset, all horses that survived had returned to an exercise level comparable to that experienced before the outbreak. Factors associated with the development of neurologic disease included age of > 5 years, location in the south or arena stall areas, and highest rectal temperature on day 3 or later of the febrile period. CONCLUSIONS AND CLINICAL IMPORTANCE: Being > 5 years of age, having had a rectal temperature of > 103.5 degrees F, and highest rectal temperature occurring on or after the 3rd day of the febrile period were the factors most predictive of the development of neurologic disease and death. Data obtained during this outbreak substantiate previous findings relating to clinical aspects and diagnosis of EHV-1 myeloencephalopathy. The prophylactic and therapeutic use of acyclovir during this outbreak is described.

Equine herpesvirus 1 and 4.

Equine herpesvirus infections in horses remain a significant cause of abortion and neurologic disease. These viruses are also responsible for mild signs of respiratory disease. The ability to establish latent infections with periodic reactivation or transmission to other horses is an important feature of these herpesviruses. One of the most unique aspects of this report is the description of horses demonstrating neurologic signs serving as the source of infection for other horses. Accurate diagnosis and better means of protection for horses remain problems facing veterinarians and horse owners.

Rapid sequential changeover of expressed p44 genes during the acute phase of Anaplasma phagocytophilum infection in horses.

Anaplasma phagocytophilum immunodominant polymorphic major surface protein P44s have been hypothesized to go through antigenic variation, but the within-host dynamics of p44 expression has not been demonstrated. In the present study we investigated the composition and changes of p44 transcripts in the blood during the acute phase of well-defined laboratory A. phagocytophilum infections in naive equine hosts. Three traveling waves of sequential population changeovers of the p44 transcript species were observed within a single peak of rickettsemia of less than 1 month. During the logarithmic increase, the rapid switch-off of the initial dominant transcript p44-18 occurred regardless of whether the bacterium was transmitted by ticks or by intravenous inoculation. Each of the subsequently dominant p44 transcript species was phylogenetically dissimilar from p44-18. Development of antibody to the hypervariable region of P44-18 during the rickettsemia suggests the suppression of dominance of immuno-cross-reactive p44 populations. When A. phagocytophilum was preincubated with plasma from the infected horse and then coincubated with HL-60 cells, the dominance of the p44-18 transcript was rapidly suppressed in vitro and most of the newly emerged p44 transcript species were previously undetected in this horse. This work provides experimental evidence of within-host p44 antigenic variation. Results suggest that the rapid and synchronized switch of expression is an intrinsic property of p44s reinitiated after transmission to naive mammalian hosts and shaped upon exposure to immune plasma.

Technique for equine cervical articular process joint injection.

Degenerative changes and osteochondrosis of articular processes are common sources of stiffness or pain in the equine cervical spine. Temporary relief of the clinical signs related to these maladies may be achieved by injecting corticosteroids into the joint. This is routinely done by ultrasound-guided needle placement. The cervical articular processes and joint form an easily identifiable sonographic landmark, a step-like echogenic surface described as having the appearance of a "chair."

Evaluation and comparison of an indirect fluorescent antibody test for detection of antibodies to Sarcocystis neurona, using serum and cerebrospinal fluid of naturally and experimentally infected, and vaccinated horses.

The objectives of this study were to evaluate the accuracy of the indirect fluorescent antibody test (IFAT) using serum and cerebrospinal fluid (CSF) of horses naturally and experimentally infected with Sarcocystis neurona, to assess the correlation between serum and CSF titers, and to determine the effect of S. neurona vaccination on the diagnosis of infection. Using receiver-operating characteristic analysis, the areas under the curve for the IFAT were 0.97 (serum) and 0.99 (CSF). Sensitivity and specificity were 83.3 and 96.9% (serum, cutoff 80) and 100 and 99% (CSF, cutoff 5), respectively. Titer-specific likelihood ratios (LRs) ranged from 0.03 to 187.8 for titers between <10 and 640. Median time to conversion was 22-26 days postinfection (DPI) (serum) and 30 DPI (CSF). The correlation between serum and CSF titers was moderately strong (r = 0.6) at 30 DPI. Percentage of vaccinated antibody-positive horses ranged from 0 to 95% between 0 and 112 days after the second vaccination. Thus, the IFAT was reliable and accurate using serum and CSF. Use of LRs potentially improves clinical decision making. Correlation between serum and CSF titers affects the joint accuracy of the IFAT; therefore, the ratio of serum to CSF titers has potential diagnostic value. The S. neurona vaccine could possibly interfere with equine protozoal myeloencephalitis diagnosis.