Prediction of serum ionized calcium concentration by use of serum total calcium concentration in dogs

OBJECTIVE: To determine whether total serum calcium (tCa) or adjusted tCa concentrations accurately predict ionized calcium (iCa) status in dogs. SAMPLE POPULATION: 1,633 canine serum samples. PROCEDURE: The tCa concentration was adjusted for total protein (TP) or albumin concentration by use of published equations. Correlations between iCa and tCa or adjusted tCa, tCa and TP, and tCa and albumin were calculated. Diagnostic discordance between tCa or adjusted tCa and iCa was determined. Diagnostic discordance in predicting iCa was also determined for 490 dogs with chronic renal failure (CRF). Sensitivity, specificity, positive and negative predictive values, and positive and negative diagnostic likelihood ratios were calculated for tCa, tCa adjusted forTP, and tCa adjusted for albumin. RESULTS: Diagnostic discordance was 27% when tCa concentration was used to predict iCa status. Use of adjusted tCa increased diagnostic discordance to approximately 37% for all dogs and 55% for dogs with CRF. Positive predictive value and positive diagnostic likelihood ratios were poor when tCa concentration was used to predict iCa status. The tCa concentration overestimated normocalcemia and underestimated hypocalcemia. Adjusted tCa overestimated hypercalcemia and underestimated hypocalcemia. CONCLUSIONS AND CLINICAL RELEVANCE: Adjusted tCa or tCa concentrations are unacceptable for predicting iCa status in dogs. Use of adjustment equations is not recommended. Direct measurement of iCa concentration is necessary for accurate assessment of calcium status. Use of tCa or adjusted tCa concentrations to predict iCa status in dogs could cause serious mistakes in diagnosis and case management, especially in dogs with CRF.     

Prediction of serum ionized calcium concentration by serum total calcium measurement in cats

Feline serum samples (n = 434) were classified as hypercalcemic, normocalcemic, or hypocalcemic based on both total calcium (tCa) and ionized calcium (iCa) concentrations. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive diagnostic likelihood ratio (PDLR), and negative diagnostic likelihood ratio (NDLR) were calculated for prediction of hypercalcemia and hypocalcemia in all samples, in hypoalbuminemic cats, and in those with chronic renal failure (CRF) as compared with cats that had other conditions. Diagnostic discordance in prediction of iCa using tCa was 40%. Sensitivity of tCa in prediction of ionized hypercalcemia was low and specificity was high. The PDLR for prediction of ionized hypercalcemia or hypocalcemia was low in all cats, especially in those with CRF. Due to the high level of diagnostic discordance, tCa should not be used to predict iCa concentration. Concentration of iCa should be measured directly when accurate assessment of calcium status is needed.     

Determination of serum ionized calcium concentration in dairy cattle after frozen anaerobic storage

The stability of serum ionized calcium concentration (ICa) from dairy cows was studied after anaerobic collection and frozen storage. Paired blood samples were obtained from five groups of cows: nonlactating, first third of lactation, midlactation, last third of lactation, and 2-year-old nonlactating heifers. Vacutainer multiple sample needles and serum separator tubes (SST) were used for venipuncture. Aspiration of serum was within 1.5 hours after collection: one sample for immediate determination (within 2 hours of collection); the other sample stored at -4 degrees C in evacuated plastic vacutainer tubes filled with serum to provide dead space of less than 75% of volume, and analyzed after 14 to 30 days in storage (half, 15, of the samples from each lactation group were analyzed after frozen anaerobic storage at 14 or 30 days, respectively). Processing samples in this manner significantly altered the values obtained for ICa, normalized calcium concentration (NCa), and pH. Analysis after frozen storage in evacuated tubes caused ICa and NCa concentrations to decrease and pH to increase (P< 0.05); total calcium levels were not significantly different from initial values. There were no significant differences among lactation groups. The difference between values obtained from these paired samples was either due to loss of CO(2) during transfer from the SST to the evacuated tube or during frozen storage. Changes in samples assayed after freezing and storage could be adjusted to original values by using the mean difference between the fresh and frozen levels as correction factors: ICa (+0.4379), NCa (+0.2797), and pH (-0.0926). It was concluded that immediate determination of serum ICa in dairy cattle is the ideal but using this methodology and performing analyses later may be acceptable if correction factors are determined.     

Plasma concentration of ionized calcium in healthy iguanas

OBJECTIVE: To measure plasma concentration of ionized calcium in healthy green iguanas. DESIGN: Prospective study. ANIMALS: 9 juvenile and 21 (10 male, 11 female) adult iguanas. PROCEDURE: Blood samples were obtained from each iguana, and plasma calcium, glucose, phosphorus, uric acid, total protein, albumin, globulin, potassium, and ionized calcium concentrations, aspartate transaminase (AST) activity, and pH were measured. Heparinized blood was used for measurement of ionized calcium concentration and blood pH. A CBC was also performed to assess the health of the iguanas. RESULTS: Significant differences were not detected among the 3 groups (juveniles, males, and females) with regard to ionized calcium concentration. Mean ionized calcium concentration measured in blood was 1.47 +/- 0.105 mmol/L. Significant differences were detected between juveniles and adults for values of phosphorus, glucose, total protein, albumin, globulin, and AST activity. CONCLUSIONS AND CLINICAL RELEVANCE: Ionized calcium concentration provides a clinical measurement of the physiologically active calcium in circulation. Evaluation of physiologically active calcium in animals with suspected calcium imbalance that have total plasma calcium concentrations within reference range or in gravid animals with considerably increased total plasma calcium concentrations is vital for determining a therapeutic plan. Accurate evaluation of calcium status will provide assistance in the diagnosis of renal disease and seizures and allow for better evaluation of the health status of gravid female iguanas.     

Effects of storage on serum ionized calcium and pH from horses with normal and abnormal ionized calcium concentrations

It has been previously shown that Ca(I) concentration is stable in serum collected from healthy horses for 10 days if stored at 40 degrees C. This may not be true for horses with abnormal Ca(I) concentrations. Thus the stability of ionized calcium (Ca(I)) concentration and pH measurement in serum from horses with both normal and abnormal Ca(I) concentrations stored for various times at 40 degrees C and -10 degrees C was evaluated. Our results indicated that serum Ca(I) concentration was stable throughout 7 days of cold or frozen storage, after being received by the Clinical Chemistry Laboratory. Serum Ca(I) concentration showed a significant decrease by 14 days of frozen storage (-10 degrees C). Serum pH showed a statistically significant increase by 7 days of cold storage, and within 3 days of frozen storage. If equine serum is collected, handled and stored anaerobically, and kept cold or frozen, Ca(I) concentration can be accurately measured for approximately 7 days after collection, regardless of the health status of the animal. An accurate measurement of pH may be made within 3 days of cold or 1 day of frozen storage.     

Serum ionized calcium concentration in clinically normal dairy cattle, and changes associated with calcium abnormalities

Serum ionized calcium (ICa) concentration was determined in 141 clinically normal dairy cattle by use of a direct-measuring calcium ion-selective electrode instrument. Mean serum ICa concentration 2 hours after blood withdrawal was 4.59 mg/dl; range varied from 3.79 to 5.25 mg/dl. Regression analysis indicated a high degree of correlation between ICa and serum total calcium concentrations if serum stored at 23 C was analyzed within 12 hours after blood withdrawal. Abnormal ICa concentration was detected in 19 of 85 dairy cows that were affected with various pathologic conditions. All 19 cows had hypocalcemia (n = 13 with parturient hypocalcemia, 4 with hypomagnesemic tetany, and 2 with renal disease). In all cases, the ICa concentration clearly related to the clinical manifestation of disease and the functional status of the cow's calcium metabolism.     

Effect of sodium bicarbonate infusions on ionized calcium and total calcium concentrations in serum of clinically normal cats

The effects of sodium bicarbonate (0.5 mEq/kg of body weight, 1.0 mEq/kg, 2.0 mEq/kg, and 4.0 mEq/kg) on ionized and total calcium concentrations were determined in clinically normal cats. Also, serum pH, whole blood pH, and serum albumin, serum total protein, and serum phosphorus concentrations were measured. Intravenous administration of sodium bicarbonate to awake cats decreased serum ionized calcium and serum total calcium concentrations. All dosages of sodium bicarbonate were associated with significant decreases of serum ionized calcium concentration. This effect lasted for greater than 180 minutes when cats were given 2.0 mEq/kg or 4.0 mEq/kg. When cats were given 4 mEq of sodium bicarbonate/kg, serum ionized calcium concentration was significantly decreased, compared with that when cats were given lower doses, but only at 10 minutes after infusion. After sodium bicarbonate infusion, serum total calcium concentration, measured by ion-specific electrode and colorimetry, was lower than baseline values at most of the times evaluated. Decreases in serum ionized calcium and serum total calcium concentrations can be attributed only in part to an increase in serum or whole blood pH and to a decrease in serum protein concentration. Serum total calcium concentrations measured by ion-specific electrode and by colorimetry were positively correlated, but the variability was high. Only 44% of the variability in serum ionized calcium concentration could be predicted when serum total calcium, albumin, total protein, phosphorus, and bicarbonate concentrations and pH were considered.     

The concentration of ionized magnesium in serum during the periparturient period of non-paretic dairy cows

Ion-selective electrodes have recently been designed for determining the ionized concentration of magnesium (Mg²⁺) in serum. This development may allow new insights into some metabolic diseases of cattle. For this report, the concentrations of Mg²⁺, total magnesium (Mg_tot), ionized calcium (Ca²⁺), total calcium (Ca_tot), and inorganic phosphate (P_i) were determined in sera from seventeen 3-to 16-year-old Brown Swiss and crossed Simmental/Red Holstein cows during the periparturient period. In each animal, a transient increase of Mg²⁺ and Mg_tot serum concentrations was observed in association with the transient decrease in serum concentrations of Ca²⁺, Ca_tot and P_i after parturition. On average, throughout the study, the serum Mg²⁺ concentrations were 68.5% of those of Mg_tot, whereas the serum Ca²⁺ concentrations were 52% of those of Ca_tot. The possible mechanisms involved in the transient increase of Mg²⁺ and Mg_tot serum concentrations are discussed.Abbreviations Ca²⁺ ionized calcium - Ca_tot total calcium - Mg²⁺ ionized magnesium - Mg_tot total magnesium - P_i inorganic phosphate - PTH parathyroid hormone - PTHrP parathyroid homrone related protein     

Validation of the bovine blood calcium checker as a rapid and simple measuring tool for the ionized calcium concentration in cattle

Point-of-care (POC) devices that veterinary practitioners can use to easily and rapidly measure blood ionized calcium (iCa) levels in cows immediately after withdrawing a blood sample on the dairy farm are needed. Aims of present studies was to compare the commercially available ion-selective electrode handheld iCa meter (bovine blood iCa checker) with the benchtop blood gas analyzer GEM premier 3500 and handheld analyzer i-STAT 1. Sixty-two paired-point whole blood samples were obtained from three cows with hypocalcemia experimentally induced by Na₂-EDTA infusion. Whole blood samples were also obtained from the 36 cows kept on a farm in field conditions. The results using the bovine blood iCa checker correlated with those using the GEM premier 3500 and i-STAT 1. Bovine blood iCa checker was “compatible” with the GEM premier 3500 and i-STAT 1 because the frequency of differences between the measurements within ± 20% of the mean were 100% (65/65, >75%) and 90.8% (59/65, >75%), respectively. In the field trial, the blood iCa concentration measured by the bovine blood Ca checker was significantly positively correlated with that measured by the i-STAT 1 portable analyzer. Bovine blood iCa checker was “compatible” with the i-STAT 1 because the frequency of differences between the measurements within ± 20% of the mean was 100% (36/36, >75%). Results from these findings, the bovine blood iCa checker may be applied as a simplified system to measure the iCa concentration in bovine whole blood.     

Newcastle disease antibody titre is dependent on serum calcium concentration

Chickens were fed diets having optimal, high, and low levels of calcium for 42 days. Serum samples were collected at 14, 28 and 42 days of age, and serum calcium and haemagglutination inhibition titres for Newcastle disease virus were measured. The chickens were vaccinated at 14 days for Newcastle disease. Antibody titres were significantly increased by high dietary calcium and depressed by low dietary calcium. Mean titre was 2.5 (log2) for the optimal diet, 3.2 for the high-calcium diet, and 1.6 for the low-calcium diet. Antibody titres were dependent on serum calcium concentration (r2 = 0.98 at 14 days, 0.99 at 28 days, and 0.78 at 42 days).     

Studies on ionized calcium in serum and plasma from normal cows. Its relation to total serum calcium and the effects of sample storing.

Ionized calcium has been determined with a new improved instrument on serum samples from 111 Swedish red-and-white cows. Simultaneous sampling of plasma and serum was performed in 32 cows for comparison of the ionized calcium level. Multiple sampling of plasma and serum from seven cows was performed to evaluate the effect of storage at 4°C and room temperature.The normal range for ionized calcium found in this study implies that the ionized calcium fraction comprises for 43.4 ± 3.0 % (mean ± 2 s) of the total serum calcium. Simultaneous analyses on plasma and serum revealed that the plasma level of ionized calcium was generally 0.ι05 mιmol/1 lower than the serum value. pH changes in stored blood samples have a direct effect on the ionized calcium levels and is therefore to be avoided. Storing samples in vacutainers for five days at 4°C or for two days at room temperature was accompanied by only small decreases of serum or plasma ionized calcium.The new instrument used in this study enables rapid analyses, and most of earlier drawbacks with calcium-ion-selective analyzers have been eliminated.     

Effects of intravenous administration of two volumes of calcium solution on plasma ionized calcium concentration and recovery from naturally occurring hypocalcemia in lactating dairy cows

OBJECTIVE: To compare the effects of administration of 2 volumes of a calcium solution (calcium oxide and calcium gluconate) on plasma ionized calcium concentration (PICaC) and clinical recovery from naturally occurring hypocalcemia (NOHC; milk fever) in lactating dairy cows. ANIMALS: 123 cows with NOHC (PICaC < 0.95 mmol/L [3.81 mg/dL]) and 20 clinically normal control cows. PROCEDURES: Affected cows were treated IV once or repeatedly with 450 (n = 56) or 750 mL (67) of calcium solution (1.65 g of calcium/100 mL) until clinical recovery was achieved. The PICaC was assessed 48 hours after the first treatment or after the treatment that achieved clinical recovery. Biochemical recovery was defined as PICaC > or = 0.95 mmol/L. Plasma from control cows was used for PICaC reference range determination. Plasma samples from both groups were assessed after storage for 20 days at 20 degrees C. RESULTS: The PICaC reference range derived from blood collected in tubes containing lithium heparin was 1.02 to 1.29 mmol/L (4.09 to 5.17 mg/dL). Following storage, plasma samples were suitable for PICaC assessment. All cows treated with > or = 1 volume of 450 and 750 mL of calcium solution recovered clinically; however, 31 of 83 (37%) evaluated cows were not biochemically recovered at 48 hours following treatment. Only cows with PICaC < 0.48 mmol/L (1.92 mg/dL) before the first treatment had to be treated > or = 3 times. CONCLUSIONS AND CLINICAL RELEVANCE: Results did not support the need to increase the administered volume of calcium solution from 450 to 750 mL for treatment of NOHC in dairy cows.     

Use of a water hardness test kit to measure serum calcium concentration in cattle

OBJECTIVE: To determine whether a commercially available water hardness test kit could be used to measure total serum calcium concentration and diagnose hypocalcemia in dairy cows. DESIGN: Prospective study. ANIMALS: 30 dairy cows from 19 commercial herds. PROCEDURE: Serum calcium concentration was determined using a water hardness test kit and a standard, laboratory-based method. Simple linear regression was used to determine whether there was a linear relationship between results of the 2 methods, and Spearman's rank correlation was used to calculate correlation between measurements. Sensitivity, specificity, and predictive values of using test kit-derived values for diagnosis of hypocalcemia (laboratory value < 8 mg/dl) were calculated. RESULTS: There was a high correlation and significant linear relationship between results of the 2 methods. Sensitivity, specificity, predictive value of a positive test result, and predictive value of a negative test result were 100, 73, 86, and 100%, respectively. Accuracy was improved by using a test kit-derived calcium concentration of 7 mg/dl as the cut-off for determining hypocalcemia. CLINICAL IMPLICATIONS: Results indicate that a commercially available water hardness test kit can be used as a rapid, inexpensive method of estimating serum calcium concentrations and diagnosing hypocalcemia in dairy cattle. However, the test is not practical for cow-side use, because blood samples must be centrifuged to obtain serum for use in the test kit.     

Comparison of serum parathyroid hormone and ionized calcium and magnesium concentrations and fractional urinary clearance of calcium and phosphorus in healthy horses and horses with enterocolitis

OBJECTIVE: To evaluate calcium balance and parathyroid gland function in healthy horses and horses with enterocolitis and compare results of an immunochemiluminometric assay (ICMA) with those of an immunoradiometric assay (IRMA) for determination of serum intact parathyroid hormone (PTH) concentrations in horses. ANIMALS: 64 horses with enterocolitis and 62 healthy horses. PROCEDURES: Blood and urine samples were collected for determination of serum total calcium, ionized calcium (Ca2+) and magnesium (Mg2+), phosphorus, BUN, total protein, creatinine, albumin, and PTH concentrations, venous blood gases, and fractional urinary clearance of calcium (FCa) and phosphorus (FP). Serum concentrations of PTH were measured in 40 horses by use of both the IRMA and ICMA. RESULTS: Most (48/64; 75%) horses with enterocolitis had decreased serum total calcium, Ca2+, and Mg2+ concentrations and increased phosphorus concentrations, compared with healthy horses. Serum PTH concentration was increased in most (36/51; 70.6%) horses with hypocalcemia. In addition, FCa was significantly decreased and FP significantly increased in horses with enterocolitis, compared with healthy horses. Results of ICMA were in agreement with results of IRMA. CONCLUSIONS AND CLINICAL RELEVANCE: Enterocolitis in horses is often associated with hypocalcemia; 79.7% of affected horses had ionized hypocalcemia. Because FCa was low, it is unlikely that renal calcium loss was the cause of hypocalcemia. Serum PTH concentrations varied in horses with enterocolitis and concomitant hypocalcemia. However, we believe low PTH concentration in some hypocalcemic horses may be the result of impaired parathyroid gland function.     

Effect of changes in ionized calcium concentration in arterial blood and metabolic acidosis on the arterial partial pressure of oxygen in dogs

OBJECTIVE: To evaluate the effects of metabolic acidosis and changes in ionized calcium (Ca2+) concentration on PaO2 in dogs. ANIMALS: 33 anesthetized dogs receiving assisted ventilation. PROCEDURE: Normal acid-base status was maintained in 8 dogs (group I), and metabolic acidosis was induced in 25 dogs. For 60 minutes, normocalcemia was maintained in group I and 10 other dogs (group II), and 10 dogs were allowed to become hypercalcemic (group III); hypocalcemia was then induced in groups I and II. Groups II and IV (5 dogs) were treated identically except that, at 90 minutes, the latter underwent parathyroidectomy. At intervals, variables including PaO2, Ca2+ concentration, arterial blood pH (pHa), and systolic blood pressure were assessed. RESULTS: In group II, PaO2 increased from baseline value (96 +/- 2 mm Hg) within 10 minutes (pHa, 7.33 +/- 0.001); at 60 minutes (pHa, 7.21 +/- 0.02), PaO2 was 108 +/- 2 mm Hg. For the same pHa decrease, the PaO2 increase was less in group III. In group I, hypocalcemia caused PaO2 to progressively increase (from 95 +/- 2 mm Hg to 104 +/- 3 mm Hg), which correlated (r = -0.66) significantly with a decrease in systolic blood pressure (from 156 +/- 9 mm Hg to 118 +/- 10 mm Hg). Parathyroidectomy did not alter PaO2 values. CONCLUSIONS AND CLINICAL RELEVANCE: Induction of hypocalcemia and metabolic acidosis each increased PaO2 in anesthetized dogs, whereas acidosis-induced hypercalcemia attenuated that increase. In anesthetized dogs, development of metabolic acidosis or hypocalcemia is likely to affect ventilatory control.