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Abstract

Objective—To determine serum lactoferrin concentrations (SLFC) in neonatal calves before and after ingestion of colostrum and to develop models that predict SLFC as a function of colostral lactoferrin concentrations (CLFC) in calves.

Animals—13 Holstein calves.

Procedure—Calves were fed 4 L of colostrum via oroesophageal feeder within 3 hours after birth. Serum samples were collected before ingestion of colostrum (day 0) and 2, 4, 6, and 7 days after birth. Colostrum and serum IgG concentrations were measured by use of radial immunodiffusion. The CLFC and SLFC were determined by use of an ELISA.

Results—Mean ± SD SLFC on days 0, 2, 4, 6, and 7 were 2.5 ± 1.6 (range 0.47 to 7.1), 6.0 ± 3.0 (range 2.0 to 16.6), 12.0 ± 12.4 (range 0.0 to 43.5), 17.1 ± 13.6 (range 2.2 to 39.4), and 13.6 ± 16.4 (range 0.0 to 43.8) mg/ml, respectively. The SLFC on days 6 and 7 differed significantly from SLFC on day 0. The model that best estimated SLFC on day 6 predicted that (SLFC)2 was a function of the logarithm of relative efficiency of passive transfer (REPT) and ([CLFC]2 × [REPT]2), where R 2 = 0.4. The model for SLFC on day 7 predicted that (SLFC)2 was a function of log(REPT), where R 2 = 0.44.

Conclusions and Clinical Relevance—Definitive evidence for passive transfer of lactoferrin via colostrum is lacking, because SLFC on day 2 or 4 were not significantly different than day 0. Relative efficiency of lactoferrin absorption was directly related to SLFC on day 6 but inversely related to SLFC on day 7. (Am J Vet Res 2002;63:476–478)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate diagnostic utility of a commercially available immunoassay for assessing adequacy of passive transfer of immunity in neonatal calves.

Design—Prospective study.

Animals—123 calves.

Procedure—Blood and serum samples were obtained from the calves prior to 2 weeks of age. The immunoassay was performed, along with refractometry and an 18% sodium sulfite turbidity test. Serum IgG concentration was determined with a radial immunodiffusion assay. Sensitivity and specificity of the immunoassay, refractometry, and the sodium sulfite test were calculated by comparing results with results of the radial immunodiffusion assay.

Results—Sensitivity and specificity of the blood IgG immunoassay were 0.93 and 0.88, respectively, compared with 1.00 and 0.53 for the sodium sulfite test. For refractometry, sensitivity and specificity were 0.71 and 0.83, respectively, when a serum total solids concentration of 5.2 g/dl was used as the cutoff between positive and negative test results.

Conclusions and Clinical Relevance—Results suggest that the immunoassayperforms well in detecting calves with inadequate passive transfer of immunity. (J Am Vet Med Assoc 2002;220:791–793)

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine whether serum IgG concentrations in neonatal calves are adversely affected by short-term frozen storage of colostrum.

Design—Prospective study.

Sample Population—Experiment 1 consisted of 10 pairs of Holstein calves (n = 20) fed matched aliquots of either fresh (n = 10) or frozen and thawed (10) colostrum. In experiment 2, 26 Holstein calves were fed either fresh (n = 13) or frozen and thawed (n = 13) colostrum.

Procedure—Experiment 1 consisted of calves resulting from observed parturitions; calves were randomly assigned to treatment groups (fresh or frozen and thawed colostrum) in pairs. Calves were fed 4 L aliquots of colostrum via oroesophageal intubation at 3 hours of age. Serum IgG concentrations at 2 days of age were compared between the 2 groups by use of a paired t-test. Experiment 2 consisted of calves resulting from observed parturitions; calves were randomly assigned to treatment groups (fresh or frozen and thawed colostrum). Calves were fed 4 L aliquots of colostrum via oroesophageal intubation at 3 hours of age. Regression analysis was used to determine whether calf serum IgG concentration was a function of colostral IgG concentration and colostrum storage group.

Results—Significant differences were not observed between the 2 groups in experiment 1. No significant relationship was observed between colostrum storage group and serum IgG concentration in experiment 2. The model that best predicted serum IgG concentrations accounted for 20% of the variability in serum IgG concentration.

Conclusion and Clinical Relevance—Frozen colostrum is an adequate source of IgG for calves. (J Am Vet Med Assoc 2001;219:357–359)

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To characterize gelatinases in bronchoalveolar lavage fluid (BALF) and gelatinases produced by alveolar macrophages of healthy calves.

Sample Population—Samples of BALF and alveolar macrophages obtained from 20 healthy 2-month-old calves.

Procedure—BALF was examined by use of gelatin zymography and immunoblotting to detect gelatinases and tissue inhibitor of metalloproteinase (TIMP)-1 and -2. Cultured alveolar macrophages were stimulated with lipopolysaccharide (LPS), and conditioned medium was subjected to zymography. Alveolar macrophage RNA was used for reverse transcriptasepolymerase chain reaction assay of matrix metalloproteinases (MMPs), cyclooxygenase-2, and inducible nitric oxide synthase.

Results—Gelatinolytic activity in BALF was evident at 92 kd (14/20 calves; latent MMP-9) and 72 kd (18/20; latent MMP-2). Gelatinolytic activity was evident at 82 kd (10/20 calves; active MMP-9) and 62 kd (17/20; active MMP-2). Gelatinases were inhibited by metal chelators but not serine protease inhibitors. Immunoblotting of BALF protein and conditioned medium confirmed the MMP-2 and -9 proteins. Endogenous inhibitors (ie, TIMPs) were detected in BALF from all calves (TIMP-1) or BALF from only 4 calves (TIMP-2). Cultured alveolar macrophages expressed detectable amounts of MMP-9 mRNA but not MMP-2 mRNA.

Conclusions and Clinical Relevance—Healthy calves have detectable amounts of the gelatinases MMP-2 and -9 in BALF. Endogenous inhibitors of MMPs were detected in BALF (ie, TIMP-1, all calves; TIMP-2, 4 calves). Lipopolysaccharide-stimulated alveolar macrophages express MMP-9 but not MMP-2 mRNA. The role of proteases in the pathogenesis of lung injury associated with pneumonia has yet to be determined. (Am J Vet Res 2004;65:163–172)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate the elimination kinetics of chlorhexidine in milk when used as an intramammary infusion to stop lactation in cows.

Design—Prospective study.

Animals—6 cows.

Procedure—The study was performed in 2 phases. Three cows were studied in each phase. All cows were treated with chlorhexidine suspension by infusion into a mastitic mammary gland quarter after 2 milkings 24 hours apart. Foremilk samples (100 mL) were collected from treated and untreated (controls) mammary gland quarters of each cow. Chlorhexidine was extracted from raw milk, and residue concentrations were quantified by use of high-performance liquid chromatography. Foremilk samples from days 2, 5, and 8 were analyzed in phase I, and samples from time 0 and days 3, 7, 14, 21, 28, 35, and 42 were analyzed in phase II.

Results—In phases I and II, there was no quantifiable transference of chlorhexidine to milk in untreated mammary gland quarters. Measurable chlorhexidine residues were found in milk from treated mammary gland quarters of 2 cows throughout the 42-day sample period in phase II. Estimated mean elimination half-life for chlorhexidine in milk was 11.5 days.

Conclusions and Clinical Relevance—On the basis of the long elimination half-life of chlorhexidine in milk from treated mammary gland quarters, the lack of human dietary exposure data to suggest a food tolerance for chlorhexidine in food products, and the Food and Drug Administration's published zero tolerance for chlorhexidine in uncooked edible calf tissues, we do not recommend extralabel use of chlorhexidine suspension as a treatment to stop lactation in mastitic mammary gland quarters of cows. (J Am Vet Med Assoc 2003; 222:1746–1749)

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine the relationship between serum and liver copper concentrations and evaluate serum copper determination for diagnosis of copper deficiency in juvenile beef calves.

Design—Cross-sectional study.

Animals—105 juvenile beef calves.

Procedure—Copper concentrations were measured in paired liver and serum samples from 6- to 9-monthold beef calves. Regression models that predicted liver copper concentration as a function of serum copper concentration were developed. Sensitivity and specificity of serum copper concentration for detection of low liver copper concentration were determined, using a range of serum copper concentrations as test endpoints. Positive and negative predictive values were calculated.

Results—The association between serum and liver copper concentrations was significant; however, regression models accounted for only a small portion of the variation in liver copper concentrations. For a serum copper concentration endpoint of 0.45 µg/g, sensitivity and specificity for detection of low liver copper concentration were 0.53 and 0.89, respectively. Positive and negative predictive values of serum copper concentration for detection of low liver copper concentration ranged from 0.37 to 0.85 and 0.63 to 0.94, respectively.

Conclusions and Clinical Relevance—Regression models are inappropriate for predicting copper status as a function of serum copper concentration. Serum copper concentration is fairly specific for detection of low liver copper concentration but only marginally sensitive when serum copper concentration of 0.45 µg/g is used as a test endpoint. The value of serum copper concentration as a diagnostic indicator depends on prevalence of copper deficiency. (J Am Vet Med Assoc 2001;218:756–760)

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine the pharmacokinetic disposition of IV administered caffeine in healthy Lama spp camelids.

Animals—4 adult male alpacas and 4 adult female llamas.

Procedures—Caffeine (3 mg/kg) was administered as an IV bolus. Plasma caffeine concentrations were determined by use of high-performance liquid chromatography in 6 animals and by use of liquid chromatography-mass spectrometry in 2 llamas.

Results—Median elimination half-life was 11 hours (range, 9.3 to 29.8 hours) in alpacas and 16 hours (range, 5.4 to 17 hours) in llamas. The volume of distribution at steady state was 0.60 L/kg (range, 0.45 to 0.93 L/kg) in alpacas and 0.75 L/kg (range, 0.68 to 1.15 L/kg) in llamas. Total plasma clearance was 44 mL/h/kg (range, 24 to 56 mL/h/kg) in alpacas and 42 mL/h/kg (range, 30 to 109 mL/h/kg) in llamas.

Conclusions and Clinical Relevance—High-performance liquid chromatography and liquid chromatography-mass spectrometry were suitable methods for determination of plasma caffeine concentrations in alpacas and llamas. Plasma caffeine concentration-time curves were best described by a 2-compartment model. Elimination half-lives, plasma clearance, volume of distribution at steady state, and mean residence time were not significantly different between alpacas and llamas. Intravenous administration of caffeine at a dose of 3 mg/kg did not induce clinical signs of excitement.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine the effects of pasteurization of colostrum on serum lactoferrin concentration and neutrophil oxidative function by comparing values from calves given pasteurized (76 C, 15 minutes) colostrum versus calves given fresh frozen colostrum.

Animals—8 Holstein bull calves were used to study the effects of pasteurization of colostrum on the absorption of lactoferrin and neutrophil oxidative burst. Three additional calves were used to study the effect of exogenous lactoferrin on neutrophil oxidative burst.

Methods—Calves were fed fresh frozen or heat pasteurized colostrum (76 C for 15 minutes) via esophageal feeder within 4 hours of birth. Neutrophils were isolated from whole blood samples. Neutrophil oxidative burst was induced by phorbol ester (300 ng/ml) stimulation of cells (1 × 106 cells) at 37 C. Serum lactoferrin concentrations were compared, using immunoblot analysis. Serum IgG concentrations were determined by radial immunoassay. Comparisons were made between the use of the 2 types of colostrum in calves by measuring subsequent serum IgG and lactoferrin concentrations and neutrophil superoxide production.

Results—Serum IgG and lactoferrin concentrations increased more in calves receiving fresh frozen colostrum. Neutrophil superoxide production was higher in neutrophils prepared from calves receiving fresh frozen colostrum. Colostral lactoferrin addition to neutrophil incubations resulted in increased oxidative burst.

Conclusions and Clinical Relevance—Compared with calves given fresh frozen colostrum, calves given pasteurized colostrum had decreased serum IgG and lactoferrin concentrations and neutrophil superoxide production 24 hours after administration. These results suggest that pasteurizing bovine colostrum at 76 C for 15 minutes has substantial effects on passive transfer of proteins and neutrophil function. (Am J Vet Res 2000;61:1019–1025)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate the effect of lactoferrin on lipopolysaccharide (LPS)-induced proliferation of bovine peripheral blood mononuclear cells (PBMCs), gene expression of inflammatory mediators, and production of prostanoids in vitro.

Sample Population—PBMCs isolated from 15 Holstein bull calves.

Procedures—Mixed populations of PBMCs were isolated by differential centrifugation. Proliferation assays were conducted in 96-well plates designed to allow addition of lactoferrin (200 ng/mL) with and without LPS (1 μg/mL) in a checkerboard design. Incorporation of 3H-thymidine was used to determine proliferation of PBMCs. Prostaglandin E2 production was determined in culture-conditioned medium by use of enzyme immunoassay. Effects of lactoferrin on LPS-induced gene expression of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 were monitored by use of PCR assays.

Results—Lactoferrin supplementation significantly reduced LPS-induced incorporation of 3H-thymidine and production of prostaglandin E2 by PBMCs. Lactoferrin reduced LPS-induced expression of COX-2 and MMP-9 mRNA.

Conclusions and Clinical Relevance—Lactoferrin reduced LPS-induced cellular proliferation, inflammatory mediator gene expression, and prostaglandin E2 production by bovine PBMCs in vitro. These effects may be beneficial in reducing the impact of endotoxemia in neonates.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To assess a commercially available point-of-care assay for measurement of bovine cardiac troponin I (cTnI) concentration in blood and plasma samples.

Sample—Prepared bovine plasma standard samples with known concentrations (0 to 1.0 ng/mL) of cTnI and blood and plasma samples obtained from 28 healthy 2.5-month-old Holstein calves.

Procedures—Coefficients of variation were calculated for concentrations of cTnI in prepared standards determined with the point-of-care assay, and values were compared with the known concentrations. The cTnI concentrations in blood samples obtained from calves determined with the point-of-care assay were compared with cTnI concentrations in plasma samples obtained from those animals determined with a validated immunoassay.

Results—The coefficients of variation of cTnI concentrations determined for prepared standards by use of the point-of-care assay were low (< 20%) for standards with cTnI concentrations ≥ 0.025 ng/mL. The blood cTnI concentrations determined with the point-of-care assay were not significantly different from the plasma cTnI concentrations determined with the validated immunoassay.

Conclusions and Clinical Relevance—Results of this study indicated the point-of-care assay had high precision for determination of cTnI concentrations in most evaluated prepared bovine plasma standard samples. The point-of-care assay may be useful for determination of circulating concentrations of cTnI in cattle.

Full access
in American Journal of Veterinary Research