Measurement of serum IgG in foals by radial immunodiffusion and automated turbidimetric immunoassay

Hypogammaglobulinemia as a result of failure of transfer of passive immunity (FTPI) is an important risk factor for infectious disease in neonatal foals. The current gold standard for determining serum immunoglobulin concentrations is radial immunodiffusion (RID). The purpose of this study was to compare immunoglobulin concentrations measured by RID with those determined by an automated turbidimetric immunoassay (TIA), which has a much shorter turnaround time. Immunoglobulin concentrations were measured by both RID and TIA in serum collected from 84 neonatal foals. Sixty-seven foals had results within the linear range for both assays. Sensitivity and specificity of TIA for diagnosis of FTPI with IgG < or = 800 mg/dL were 0.81 (95% CI 0.70-0.88) and 0.86 (95% CI 0.76-0.93) and with IgG < or = 400 mg/dL were 0.63 (95% CI 0.35-0.86) and 0.92 (95% CI 0.87-0.95), respectively. A significant linear relationship was found between IgG concentrations determined by TIA and RID (TIA = 0.9511RID + 8.4354; R2 = .59, P < .0001). The coefficients of variation for between-run and within-run precision for the TIA were 2.5 and 3%, respectively. Storage of samples from 10 foals at -20 degrees C for 10-12 months resulted in a reduction in TIA-measured serum IgG concentration of -17.6% (SD = 3.7%), indicating that long-term storage of samples at -20 degrees C should be avoided. The results of this study indicate that measurement of serum IgG by TIA can be used to evaluate foals for FTPI.     

Use of Fourier-transform infrared spectroscopy for the diagnosis of failure of transfer of passive immunity and measurement of immunoglobulin concentrations in horses

BACKGROUND: The economic, accurate, and rapid screening of foals for failure of transfer of passive immunity (FPT) is essential to ensure timely intervention. HYPOTHESIS: Infrared (IR) spectroscopy of foal sera and pattern recognition may be used to diagnose FPT and quantify serum IgG. SAMPLES: Sera from 194 foals (24-72 hours) with serum immunoglobulin G (IgG) concentrations determined previously by radial immunodiffusion assay (RID) were used. METHODS: IR spectra were recorded for the serum samples, and the data were randomly divided into training and independent test sets, each containing both FPT-positive (IgG <400 mg/dL) and non-FPT samples. A genetic optimal region selection algorithm and linear discriminant analysis were used to partition the training spectra, and the resulting classifier was then validated by comparing the IR-predicted FPT status for each of the test samples to that provided by the RID IgG assay. A quantitative IR-based assay for IgG was developed using partial least squares (PLS) and validated by testing its ability to predict IgG concentrations. RESULTS: Specificity, sensitivity, and accuracy for the combined data were 92.5, 96.8, and 95.9%, respectively. Corresponding positive (88.1%) and negative predictive (98.0%) values determined a success rate of 95-97% as compared to RID-based IgG concentrations. The IR-based quantitative assay yielded correlation coefficients for IR spectroscopy versus RID-based IgG concentrations of 0.90 and 0.86 for the training and test sets, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: The overall performance of the IR-based test was similar to that of the colorimetric assay and was superior and more economic than other available tests.     

Effect of plasma transfusion on neutrophil function in healthy and septic foals

OBJECTIVE: To evaluate the effect of plasma transfusion on phagocytosis and oxidative burst activity of peripheral blood neutrophils from healthy and septic equine neonates with sub-optimal passive transfer of maternal immunity. ANIMALS: Nine healthy and seven septic foals with suboptimal passive transfer of maternal immunity (serum IgG < 8 g/L) presented to participating veterinary hospitals for plasma transfusion, and seven healthy foals less than 7 days of age and with circulating IgG concentrations > or = 8 g/L. PROCEDURE: Foals with serum IgG concentrations < 8 g/L were assessed as healthy or septic. Sepsis was recognised by positive bacterial cultures and/or sepsis scores of > or = 11. All foals received between 1 and 3 L of plasma to boost circulating IgG concentrations to > or = 8 g/L. Serum IgG concentrations were determined before and following transfusion by glutaraldehyde coagulation test and confirmed by single radial immunodiffusion assays. Neutrophil phagocytosis and oxidative burst activity were determined before plasma transfusion and at 0 h, 12 h, 24 h, 48 h and 5 d following treatment. Neutrophil function from seven healthy foals less than 7 d of age and with circulating IgG concentrations of > or = 8 g/L was similarly evaluated on a single occasion. RESULTS: Plasma treatment significantly increased circulating IgG concentrations for healthy and septic foals. Oxidative burst activity of neutrophils from septic foals was significantly increased 5 days following treatment, relative to 0 h post treatment. Other differences were not significant but suggested a transient decrease in phagocytosis by neutrophils from healthy foals and increased phagocytosis by neutrophils from septic foals immediately following transfusion. Oxidative burst activity of neutrophils from septic foals tended to be less than that of healthy foals at all sampling times. Serum IgG concentrations were not correlated with neutrophil phagocytosis, but were correlated with oxidative burst activity. CONCLUSIONS: Plasma transfusion did not improve neutrophil function of healthy foals, suggesting that such treatment may be of equivocal benefit for healthy neonates. Conversely, improved neutrophil function was observed following treatment of septic foals, suggesting that plasma transfusion was beneficial for these foals. Oxidative burst activity of neutrophils from septic foals was lower than that of neutrophils from healthy foals and was significantly improved 5 days post treatment, when compared with values obtained immediately following treatment.     

Serum lactoferrin and immunoglobulin G concentrations in healthy or ill neonatal foals and healthy adult horses

BACKGROUND: Lactoferrin is a colostral glycoprotein with antimicrobial properties. HYPOTHESES: (1) Serum lactoferrin and immunoglobulin G (IgG) concentrations are correlated and increase in healthy foals after ingestion of colostrum; (2) compared to healthy foals, ill foals will have lower lactoferrin concentrations that correlate with their IgG concentration, neutrophil count, the diagnosis of sepsis, and survival; and (3) plasma concentrations of lactoferrin will be less than serum concentrations. ANIMALS: Healthy foals (n = 16), mature horses (n = 10), and ill foals 1-4 days old (n = 111) that were examined for suspected sepsis were used for blood collection. Colostrum was obtained from 10 healthy mares unrelated to the foals. METHODS: Blood was obtained from the healthy foals at birth and 1-3 days of age and from the ill foals at admission. Serum IgG was quantified by single radial immunodiffusion (SRID). Lactoferrin concentrations in colostrum and blood were determined by an enzyme-linked immunosorbant assay. The sepsis score, blood culture results, neutrophil counts, and survival were obtained on ill foals. RESULTS: The mean colostral lactoferrin concentration was 21.7 microg/mL. Compared to values at birth, serum IgG (18+/-2 versus 2,921+/-245 mg/dL, SEM) and lactoferrin (249+/-39 versus 445+/-63 ng/mL, SEM) concentrations were significantly greater in healthy foals 1-3 days old. Serum lactoferrin concentration in 1-3-day-old healthy foals was not different from mature horses or ill foals. IgG and lactoferrin concentrations were significantly correlated only in healthy foals. Serum lactoferrin concentrations were significantly lower in ill neutropenic foals. The serum IgG concentration was significantly lower in ill foals as compared to healthy foals. Only serum IgG was significantly less in ill foals with a positive sepsis score and in nonsurvivors, Plasma lactoferrin concentrations were lower than serum concentrations, although values were significantly correlated. CLINICAL IMPORTANCE: Although both serum IgG and lactoferrin concentrations increase in healthy foals after ingestion of colostrum, only serum IgG is significantly correlated with the sepsis score and outcome.     

Effects of oral administration of concentrated equine serum IgG to newborn foals on passive immunity

Thirteen newborn foals of Quarter Horse breeding were used to determine if oral administration of concentrated equine serum increases concentrations of IgG in foals allowed to naturally suckle colostrum. Foals were alternately assigned either to receive 300 ml of an oral equine serum IgG product or to serve as controls. Foals receiving the IgG product were given 150 ml orally at 10 hours and again at 12 hours after birth. All foals were allowed to suckle from their dams ad libitum. Jugular blood samples were obtained from foals at 10 hours and 24 hours of age for IgG determination. Colostrum samples from the dam were also obtained within 3 hours following parturition for determination of specific gravity. Plasma samples were analyzed for IgG level using a commercially available radial immunodiffusion kit. Oral administration of equine serum IgG had no significant effect on concentrations of plasma IgG in foals at 24 hours of age (p>.34). There was also no difference between control and treated foals in the rate of IgG absorption from 10-24 hours after birth (p>.34). In conclusion, oral administration of equine IgG to foals that ingest their dam's colostrum does not significantly increase concentrations of plasma IgG when compared to controls.     

Neutrophil function and plasma opsonic capacity in colostrum-fed and colostrum-deprived neonatal kittens

OBJECTIVE: To determine whether passive transfer of IgG in neonatal kittens affects plasma opsonic capacity and neutrophil phagocytic and oxidative burst responses to bacteria in vitro. ANIMALS: 22 kittens from 6 specific pathogen-free queens. PROCEDURE: Kittens were randomized at birth into the following treatment groups: colostrum-fed, colostrum-deprived, or colostrum-deprived supplemented with feline or equine IgG. Blood samples were collected at intervals from birth to 56 days of age. Plasma IgG concentrations were determined by radial immunodiffusion assay. Neutrophil function was assessed by a flow cytometry assay providing simultaneous measurement of bacteria-induced phagocytosis and oxidative burst. The opsonic capacity of kitten plasma was determined in an opsonophagocytosis assay with bacteria incubated in untreated or heat-inactivated plasma. RESULTS: Among treatment groups, there were no significant differences in neutrophil phagocytic and oxidative burst responses to bacteria or opsonic capacity of plasma. In all samples of plasma, inactivation of complement and other heat-labile opsonins significantly reduced the opsonic capacity. Plasma IgG concentrations in kittens did not correlate with neutrophil function or plasma opsonic capacity before or after inactivation of complement. CONCLUSIONS AND CLINICAL RELEVANCE: The plasma opsonic capacity and neutrophil phagocytic and oxidative burst responses in vitro of kittens receiving passive transfer of IgG via colostrum intake or IgG supplementation and those deprived of colostrum were similar. The alternate complement pathway or other heat-labile opsonins may be more important than IgG in bacterial opsonization and phagocytosis.     

Plasma vasopressin concentrations in healthy foals from birth to 3 months of age

Background: Arginine vasopressin (AVP) has received increased attention in equine critical care but there is minimal information of AVP concentration in foals. The clinical usefulness of measuring AVP in ill foals depends on knowledge of age-related changes in AVP concentrations in healthy foals.
Hypothesis: Plasma AVP concentrations will be significantly different when measured from birth to 3 months of age in healthy foals.
Animals: Thirteen healthy university-owned foals.
Methods: Prospective, observational study. Blood was collected from healthy foals at birth and 3, 5, 7, 10, 14, 21, 28, 42, 56, and 84 days of age. Plasma was harvested and plasma AVP concentrations were determined by radioimmunoassay.
Results: No statistically significant differences were detected in plasma AVP concentrations over the study period. Plasma AVP concentrations over the entire study period was 6.2 ± 2.5 pg/mL.
Conclusions and Clinical Importance: There was no age-related variation in plasma AVP concentrations detected in healthy foals from birth to 3 months of age suggesting that AVP concentrations are similar across foals of these ages.     

Evaluation of plasma carboxy-terminal cross-linking telopeptide of type I collagen concentration in horses

OBJECTIVE: To evaluate a human assay for quantification of carboxy-terminal cross-linking telopeptide of type I collagen (CTX-I), assess the influence of age on plasma CTX-I concentration, investigate the relationship between plasma CTX-I and serum osteocalcin concentrations, and determine whether concentrations of plasma CTX-I or serum osteocalcin fluctuate in circadian manner in horses. HORSES: 75 clinically normal horses. PROCEDURE: Cross-reactivity between equine serum CTX-I and CTX-I antibodies in an automated electrochemiluminescent sandwich antibody assay (ECLIA) was evaluated via a specificity test (ie, dilution test) and recovery calculation. Serum osteocalcin concentration was measured with an equine-specific osteocalcin radioimmunoassay. To analyze diurnal variations in plasma CTX-I and serum osteocalcin concentrations, blood samples were obtained hourly during a 24-hour period. RESULTS: Results of the dilution test indicated good correlation (r > 0.99) between expected serum CTX-I concentrations and measured serum CTX-I concentrations. The calculated CTX-I recovery was 97.6% to 109.9%. Plasma CTX-I and serum osteocalcin concentrations were correlated. Plasma CTX-I concentration was inversely correlated with age of the horse. No significant circadian variations in plasma CTX-I and serum osteocalcin concentrations were detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the fully automated CTX-I ECLIA can be used for evaluation of plasma and serum samples from horses and may be a useful tool to monitor bone metabolism changes. Horses in this study did not have notable diurnal fluctuations in serum osteocalcin and plasma CTX-I concentrations.     

Influence of age and breed of equid on plasma copper and zinc concentrations

Plasma ceruloplasmin activities and plasma Cu and Zn concentrations were determined in 215 clinically normal equids of various ages and breeds. Newborn foals, regardless of breed, were hypocupremic, compared with adolescent and mature horses. The mean plasma Cu concentration of newborn Standardbred-Thoroughbred (STD-TB) foals was 2.9 mumol/L, which was about one-sixth of plasma Cu concentrations of mature horses. Newborn draft-cross foals had higher (4.6 mumol/L) plasma Cu concentrations than did newborn STD-TB foals, but plasma Cu content was only one-fifth of the dams' concentrations. Draft-cross horses, regardless of age, had plasma Cu concentrations 15% to 40% higher than did STD-TB horses. Plasma Cu concentrations of Quarter Horse yearlings were similar to those in draft-cross yearlings. Plasma ceruloplasmin activities revealed a curvilinear relationship to plasma Cu concentrations. Plasma Zn concentrations of newborn and 1-week-old STD-TB foals were 30% to 80% higher than those for yearling and mature STD-TB horses. There were no differences between draft-cross neonates and their dams in plasma Zn concentration. Plasma Zn concentrations of neonatal and mature draft-cross horses were 22% higher than those obtained for all other equids. Age and breed of equid should be a consideration in interpretations of plasma Cu and Zn concentrations in equids.     

Use of Fourier-transform infrared spectroscopy to quantify immunoglobulin G concentration and an analysis of the effect of signalment on levels in canine serum

Deficiency in immunoglobulin G (IgG) is associated with an increased susceptibility to infections in humans and animals, and changes in IgG levels occur in many disease states. In companion animals, failure of transfer of passive immunity is uncommonly diagnosed but mortality rates in puppies are high and more than 30% of these deaths are secondary to septicemia. Currently, radial immunodiffusion (RID) and enzyme-linked immunosorbent assays are the most commonly used methods for quantitative measurement of IgG in dogs. In this study, a Fourier-transform infrared spectroscopy (FTIR) assay for canine serum IgG was developed and compared to the RID assay as the reference standard. Basic signalment data and health status of the dogs were also analyzed to determine if they correlated with serum IgG concentrations based on RID results.Serum samples were collected from 207 dogs during routine hematological evaluation, and IgG concentrations determined by RID. The FTIR assay was developed using partial least squares regression analysis and its performance evaluated using RID assay as the reference test. The concordance correlation coefficient was 0.91 for the calibration model data set and 0.85 for the prediction set. A Bland–Altman plot showed a mean difference of −89 mg/dL and no systematic bias. The modified mean coefficient of variation (CV) for RID was 6.67%, and for FTIR was 18.76%.The mean serum IgG concentration using RID was 1943 ± 880 mg/dL based on the 193 dogs with complete signalment and health data. When age class, gender, breed size and disease status were analyzed by multivariable ANOVA, dogs <2 years of age (p = 0.0004) and those classified as diseased (p = 0.03) were found to have significantly lower IgG concentrations than older and healthy dogs, respectively.     

Usefulness of a commercial equine IgG test and serum protein concentration as indicators of failure of transfer of passive immunity in hospitalized foals

Detection of failure of transfer of passive immunity (FTPI) is important in reducing morbidity and mortality in neonatal foals. We investigated the performance of a commercial equine IgG test (SNAP Foal IgG Test Kit) to diagnose FTPI in hospitalized foals. Furthermore, we evaluated the usefulness of serum total protein (STP) and serum globulin (SG) concentrations as indicators of FTPI. Serum IgG concentration was measured by means of the SNAP test and single radial immunodiffusion, and SG and STP concentrations were determined by means of a clinical chemistry analyzer. Subjects were 67 hospitalized foals <19 days old. The SNAP test was repeated on 37 samples from 29 foals, with identical results for 24 samples (kappa statistic, 0.64; 95% confidence interval [CI], 0.46-0.82). The sensitivity of the SNAP test to detect serum IgG concentration [IgG] < or =400 and < or =800 mg/dl was 90% (95% CI, 71-98%) and 95% (85-99%), respectively, and the specificity was 79% (71-82%) and 52% (39-57%), respectively. Sensitivity for detection of [IgG] < or =400 mg/dl was not affected (P > .05) by plasma fibrinogen concentration, sepsis score, or bacteremia. Specificity for detection of [IgG] < or = 800 mg/dl was lower (P < .05) in foals with sepsis score < or =11 (50% [31-60%] versus 100% [8-100%]) and bacteremia (25% [5-56%] versus 62% [45-62%]). Sensitivity and specificity of [STP] < or = 5.0 g/dl for [IgG] < or =800 mg/dl was 94% (83-99%) and 47% (30-56%), respectively. Performance of the SNAP test in hospitalized foals is impaired because of low specificity, but can have usefulness provided that the properties of the test and characteristics of the foal being examined are considered when interpreting the results. The STP and SG concentrations are poor sole indicators of FTPI in hospitalized foals, but may be useful adjunctive tests.     

Passive transfer failure in horses: incidence and causative factors on a breeding farm

A prospective study was performed to determine the incidence and associated maternal and managemental factors of failure of passive transfer (FPT) in foals on a breeding farm. The zinc sulfate turbidity test (ZSTT) and latex agglutination test (LAT) were compared for accuracy in estimating serum immunoglobulin (Ig)G of foals, as determined by single radial immunodiffusion (SRID). Complete past and present foaling histories of 136 Standardbred mares were obtained. All foalings were witnessed by farm attendants, and colostral samples were collected from mares within 2 hours after parturition. Foals that did not rise and nurse were supplemented with colostrum from the dam, using a bottle or nasogastric tube. Serum samples were prepared from foals and mares between 24 and 36 hours after parturition, and from some mares 45 to 90 days before parturition. Serum IgG concentrations of mares and foals and colostral whey were determined, using SRID. Serum IgG also was estimated in foals, using ZSTT and a commercially available LAT. Four of the 136 foals (2.9%) had FPT (serum IgG less than or equal to 400 mg/dl). Serum IgG concentrations in foals significantly correlated with colostral IgG (P less than 0.001). A significantly larger proportion of foals with FPT were bottle-fed their colostrum (P less than 0.01). Month of parturition, mare age, parity, number of barren seasons, incidence of assisted births or retained placenta, or prepartum serum IgG concentrations did not significantly affect colostral IgG concentrations or serum IgG concentrations in foals. As serum IgG concentrations in foals decreased and as colostral IgG concentrations decreased, the proportion of mares that prelactated significantly (P less than 0.01) increased.(ABSTRACT TRUNCATED AT 250 WORDS)     

Assessment of different methods to estimate bovine colostrum quality on farm

AIMS: To evaluate two different hydrometers and an optical and a digital Brix refractometer for the assessment of bovine colostrum quality, in terms of accuracy and precision compared with the measurement of IgG concentrations using radial immunodiffusion (RID), and to evaluate the reliability and repeatability of the Brix refractometers.

METHODS: To determine reliability and repeatability, 145 colostrum samples were tested by two independent observers twice, using the optical and digital Brix refractometers. A further 193 colostrum samples from Holstein cows were collected on one commercial dairy farm at first milking and tested with two hydrometers and an optical and digital Brix refractometer. An aliquot of each sample was frozen for RID measurement of IgG concentrations and samples were classified as poor (≤50 g IgG/L) or good (>50?g?IgG/L) quality colostrum. Intraclass correlation coefficients (ICC) were used to determine inter- and intra-observer reliability and repeatability. Optimised cut-off values for the four devices were determined using receiver operating characteristics (ROC) analysis with the RID results as the reference. Using these cut-offs, sensitivities and specificities for determining good quality colostrum were calculated.

RESULTS: The ICC for inter-observer reliability was 0.98 for the optical Brix refractometer, and for intra-observer repeatability was 0.97 and 0.98 for the optical and the digital Brix refractometers, respectively. For the 193 colostrum samples, 67 (34.7%) had concentrations of IgG ≤50 g/L determined by RID. Optimised cut-off values evaluated by ROC analysis were higher for all devices compared with manufacturer reference or previously published values. Using these values, the sensitivities for the two hydrometers, and the optical and the digital Brix refractometers were 0.73, 0.71, 0.56 and 0.79, respectively; specificities were 0.72, 0.61, 0.90 and 0.69, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE: The Brix refractometers provided the most accurate assessment of colostrum quality of the devices evaluated, and demonstrated excellent precision in terms of repeatability. To provide optimal health for newborn calves, a sufficient intake of good quality colostrum is essential. The Brix refractometers provide rapid, convenient tools for classification of colostrum quality.     

Immunoglobulin isotypes in sera and nasal mucosal secretions and their neonatal transfer and distribution in horses

OBJECTIVE: To determine concentrations of IgA and IgG subclasses in serum, colostrum, milk, and nasal wash samples of adult horses and foals. ANIMALS: Seven 2-year-old Welsh ponies, 27 adult mixed-breed horses, and 5 Quarter Horse mares and their foals. PROCEDURE: Serum was obtained from ponies and adult horses. Colostrum and milk were obtained from mares and serum and nasal wash samples from their foals immediately after parturition and on days 1, 7, 14, 28, 42, and 63. Nasal wash samples were also obtained from 23 adult horses. Concentrations of immunoglobulins were determined by use of inhibition ELISA. To determine transfer of maternal isotypes to foals, concentrations in colostrum and milk were compared with those in foal serum. Serum half-lives of isotypes in foals were also determined. RESULTS: IgGb was the most abundant isotype in serum and colostrum from adult horses, whereas IgA was the predominant isotype in milk. The major isotype in nasal secretions of adult horses and foals > or = 28 days old was IgA, but IgGa and IgGb were the major isotypes in nasal secretions of foals < or = 14 days old. Serum half lives of IgGa, IgGb, IgG(T), and IgA in foals were 176, 32, 21, and 3.4 days, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The early immunoglobulin repertoire of neonatal foals comprised IgGa, IgG(T), and IgA; endogenous synthesis of IgGb could not be detected until 63 days after birth. The restricted repertoire of immunoglobulins in foals may influence humoral immune responses to vaccination.     

Fibronectin concentration in plasma of mares and neonatal foals

Plasma fibronectin concentrations were measured in clinically healthy mares and their neonatal foals, using a modified human fibronectin competitive enzyme-linked immunosorbent assay. Ranges of plasma fibronectin were established in clinically healthy horses, and the assay was reliable and reproducible. Plasma fibronectin concentrations were similar in mares and foals, both before and after colostrum ingestion.     

Validation of a chemiluminescent enzyme immunometric assay for plasma adrenocorticotropic hormone in the dog

Background: The concentration of canine adrenocorticotropic hormone (ACTH) is usually determined by radioimmunoassay. However, chemiluminescent assay techniques have many advantages for clinical endocrine testing. Objectives: The objectives of this study were to validate a commercially available chemiluminescent assay for determination of canine ACTH concentration and to determine whether protease inhibitors are appropriate for use in the chemiluminescent assay system. Methods: Biological specificity was evaluated by treatment of 3 dogs with ovine corticotropin‐releasing hormone (CRH) followed by serial measurements of ACTH and by comparison with a previously validated immunoradiometric assay. All samples were collected both in the presence and absence of aprotinin, a protease inhibitor. The assay was further evaluated by measurement of intra‐assay precision, interassay precision, and recovery after dilution. Results: Baseline ACTH concentrations ranged from 5.6 to 15.3 pg/mL, and maximum ACTH concentrations of 158 to 1240 pg/mL were observed 30–60 minutes after CRH administration. Plasma samples collected with aprotinin had significantly lower ACTH concentrations than did samples collected without aprotinin. The intra‐assay coefficients of variance (CVs) ranged from 4.1 to 8.2%, and interassay CVs ranged from 4.6 to 14.8%. Recovery after dilution with canine plasma ranged from 93.4 to 103.0% of predicted concentration; however, inadequate recovery was observed with other diluents. There was a high correlation with the immunoradiometric assay (r= .925) but a significant negative bias (‐32.9, 95% confidence interval ?50.8 to ?14.9). Conclusions: This chemiluminescent assay is a valid technique for measurement of ACTH in canine plasma. ACTH concentration measured by chemiluminescence is lower than that measured by immunoradiometry. Aprotinin decreases the measured concentration of ACTH, and this effect should be taken into account when interpreting results. Diluents supplied with the kit should not be used for dilution of canine samples.     

Effects of prior feeding on pharmacokinetics and estimated bioavailability after oral administration of a single dose of microencapsulated erythromycin base in healthy foals

OBJECTIVE: To determine effects of prior feeding on pharmacokinetics and estimated bioavailability of orally administered microencapsulated erythromycin base (MEB) in healthy foals. ANIMALS: 6 healthy foals, 3 to 5 months old. PROCEDURE: Foals were given 2 doses of MEB (25 mg/kg of body weight, PO). One dose was administered after food was withheld overnight, and the other was administered after foals had consumed hay. The study used a crossover design with a 2-week period between doses. Blood was collected via a jugular vein prior to and at specific times after drug administration. Concentrations of erythromycin A and anhydroerythromycin A in plasma were determined, using high-performance liquid chromatography. Results pharmacokinetic analysis of plasma concentration-time data for food-withheld and fed conditions were compared. RESULTS: Plasma concentrations of erythromycin A for foals were lower after feeding than concentrations when food was withheld. Area under the plasma concentration-time curve, maximum plasma concentration, and estimated bioavailability were greater in foals when food was withheld than when foals were fed. Anhydroerythromycin A was detected in plasma after administration of MEB in all foals. CONCLUSIONS AND CLINICAL RELEVANCE: Foals should be given MEB before they are fed hay. Administration of MEB to foals from which food was withheld overnight apparently provides plasma concentrations of erythromycin A that exceed the minimum inhibitory concentration of Rhodococcus equi for approximately 5 hours. The dosage of 25 mg/kg every 8 hours, PO, appears appropriate.     

Prevalence (treatment days) and severity of illness in hypogammaglobulinemic and normogammaglobulinemic foals

Serum samples for determination of IgG concentration were obtained between postpartum hours 18 and 48 from 132 Standardbred foals. Results of the IgG assay were not known to farm personnel. None of the foals was given plasma IV for treatment of hypogammaglobulinemia. Foal health records were examined retrospectively to determine prevalence of infectious-type illness (foal treatment days [FTD]), prevalence of life-threatening infectious illness (foal treatment days-serious condition [FTD-SC]), and number of diseases (NOD) per foal. Values for FTD, FTD-SC, and NOD per foal were compiled for the first 21 days of life and for the first 90 days of life. The FTD, FTD-SC, and NOD per foal values were compared for foals with less than 400 mg of IgG/dl and for foals with greater than or equal to 400 mg of IgG/dl; the same variables were compared for foals with less than 800 mg of IgG/dl and for foals with greater than or equal to 800 mg of IgG/dl. Statistical analysis indicated that IgG concentration was not associated with FTD, FTD-SC, or NOD in foals of any of the groups. Also, despite a large subpopulation of hypogammaglobulinemic foals (13.6% with less than 400 mg of IgG/dl and 44.7% with less than 800 mg of IgG/dl), the 21-day and 90-day overall survival rates were 100 and 99.2%, respectively. The data strongly suggest that serum IgG concentration was not related to prevalence or severity of illness or to survival rate in this population of foals.     

Colostral and serum IgG, IgA, and IgM concentrations in Standardbred mares and their foals at parturition

Immunoglobulin G, IgM, and IgA concentrations were measured in serum collected from 36 Standardbred mares within 12 hours of foaling, in colostrum collected within 6 hours of foaling, and in serum collected from foals 24 to 48 hours after birth. In serum collected from mares after parturition, mean concentrations of IgG, IgM, and IgA were 2,463.9 +/- 1,337.3 mg/dl, 136.4 +/- 218 mg/dl, and 305.2 +/- 237.5 mg/dl, respectively. In serum from foals, mean concentrations of IgG, IgM, and IgA were 1,953.3 +/- 1,635 mg/dl, 33.8 +/- 30.4 mg/dl, and 58.4 +/- 42.2 mg/dl, respectively. In colostrum, mean concentrations of IgG, IgM, and IgA were 8,911.9 +/- 6,282.2 mg/dl, 957 +/- 1088.1 mg/dl, and 122.9 +/- 77.3 mg/dl, respectively. The IgG concentrations in foal serum were poorly correlated with IgG concentrations in colostrum (r = 0.462, P less than 0.01). Correlations of IgM or IgA concentrations in serum from foals with IgM or IgA concentrations in colostrum and correlations of IgG concentrations in serum from mares with those in colostrum were not significant (P less than 0.01). Of 36 foals, 1 (2.8%) had a serum IgG concentration less than 400 mg/dl. Of 36 foals monitored for 4 months, 6 developed infectious respiratory tract disease requiring antimicrobial therapy at ages varying from 55 to 113 days; these infections were probably not related to failure or partial failure of passive transfer of antibody.     

Measurement of plasma colloid osmotic pressure in normal thoroughbred neonatal foals

A normal plasma colloid osmotic pressure (COP) interval was established for foals and compared to values for adult horses. Plasma samples were obtained from 38 Thoroughbred foals that had normal findings on postfoaling examination and 10 healthy Thoroughbred adult horses. Samples were analyzed using a commercially available colloid osmometer. Fifty samples were obtained from 38 foals. Twelve foals had 2 samples taken, 1 during the 1st 24 hours of life and the 2nd between 24 and 72 hours of life. For foals with 2 samples, only 1 randomly selected value was used in group analysis. Total plasma protein, albumin, and globulin concentrations were measured on all samples from foals. The mean measured plasma COP for foals was 18.8 +/- 1.9 mm Hg for the 38 samples analyzed. Measured plasma COP did not differ significantly over the time period examined for either the 12 paired samples (P = .13) or with regression analysis of the 38 samples (P = .13). Calculation of mean COP, based on previously published quadratic equations using total protein, albumin, and globulin concentrations, underestimated mean measured foal COP values except for when total protein measured by refractometer was used in the Landis-Pappenheimer equation. In conclusion, the plasma COP interval (95% CI: 15.0 mm Hg, 22.6 mm Hg) obtained for healthy foals in this study was found to be lower than that of healthy adult Thoroughbreds (20.6 +/- 0.7 mm Hg, P = .006).