Association of results of the glutaraldehyde coagulation test with plasma acute phase protein concentrations and hematologic findings in hospitalized cows.

Introduction: The glutaraldehyde test (GAT) allows for animal-side semi-quantitative estimation of fibrinogen and gamma-globulin concentrations in blood samples of adult cattle and therefore detection of inflammatory disease conditions. However, the test has potential limitations, especially due to the latency period until sufficiently high fibrinogen and/or gamma-globulin concentrations are reached. The aim of the present study was therefore to assess the association between results of GAT with other inflammatory markers including hematologic variables, fibrinogen, plasma haptoglobin and serum amyloid A (SAA) concentrations. Methods: For the purpose of this prospective observational study, a convenience sample of 202 cows with a broad range of inflammatory and non-inflammatory clinical conditions was included. The GAT was run on EDTA blood, fibrinogen was measured using the Clauss and the heat precipitation method, and commercially available ELISA tests were used for determination of plasma haptoglobin and SAA concentrations. Results: Shortened GAT coagulation times were more closely correlated to serum globulin (rs = -0.72) than to plasma fibrinogen concentrations measured with the heat precipitation (rs = -0.64) and the Clauss method (rs = -0.70). Cows with a markedly (≤3 min) or moderately (4-6 min) shortened coagulation time had higher (p < 0.001) plasma haptoglobin and SAA concentrations than cows with a negative test result. Total leukocyte, monocyte and neutrophil concentrations did not differ significantly between groups. An identified cut-off for the GAT coagulation time of ≤14 min had a sensitivity and specificity of 54.4 and 100%, respectively, for the prediction of an inflammatory state based on clinical findings and/or increased plasma haptoglobin or SAA concentrations. Discussion: In conclusion, this study demonstrates considerable diagnostic agreement between positive GAT results and increased plasma concentrations of haptoglobin and SAA. Despite high specificity, the test lacks sensitivity in case of acute inflammatory conditions indicating that plasma acute phase protein concentrations and hematologic findings can provide additional diagnostic information if the GAT is negative.

Acid-base imbalances and the association of blood-gas variables, electrolytes, and biochemical analytes with outcome in hospitalized calves undergoing abdominal surgery.

BACKGROUND: Surgical abdominal emergencies in calves are associated with a guarded prognosis and have the potential for complex metabolic derangements including acid-base imbalances. OBJECTIVES: To perform a comprehensive analysis of acid-base status and to assess the prognostic relevance of preoperative clinicopathologic variables in calves undergoing abdominal surgery. ANIMALS: Hospital-based study samples of 535 (dataset 1; DS1) and 83 calves (dataset 2; DS2). METHODS: Retrospective (DS1) and prospective (DS2) case series. RESULTS: In DS1, acidemia (pH <7.33) was present in 49.9%, whereas alkalemia (pH >7.37) was present in 30.7% of calves. Plasma L-lactate, chloride, and serum inorganic phosphorus concentration accounted for 51.9%, 11.6% and 9.4% of the variation of venous blood pH, respectively. Classification tree analysis indicated that a negative outcome (death or euthanasia during hospitalization) was associated with venous pO2 ≤33.6 mm Hg, anion gap >18.3 and >22.9 mEq/L, serum albumin concentration ≤36.5 and ≤29.4 g/L, serum urea concentration >4.4 mmol/L, and plasma ionized calcium concentration ≤1.26 mmol/L. The area under the receiver operating characteristic curve of this model was 0.85 (95% CI: 0.82-0.89, P < .001) and the resulting sensitivity and specificity for the prediction of nonsurvival at the optimal probability cut-point of 0.5 was 89.8% and 65.7%, respectively. In DS2 the model had a similar sensitivity and specificity of 90.5% and 70%, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinicopathologic imbalances and associated changes of acid-base status are common in calves with surgical abdominal emergencies and have clinical utility for the prediction of a negative postoperative outcome.

Macrophages in dermal disease progression of phospholipase D4-deficient Fleckvieh calves.

A new gene defect in Fleckvieh calves leads to a syndrome with partial phenotype overlap with bovine hereditary zinc deficiency. A mutation in a gene encoding phospholipase D4 (PLD4), an endosomal exonuclease, causes the disorder. In mice, PLD4 activity indirectly regulates the Toll-like receptor 9 (TLR9) pathway via degradation of microbial DNA. PLD4 absence thus results in visceral macrophage activation comparable to human macrophage activation syndrome. In this study, disease progression and the role of macrophages in affected calves were monitored clinically, clinicopathologically, and histologically over time. Breeding data identified 73 risk matings of heterozygous carriers resulting in 54 potentially PLD4-deficient calves born on farms. PLD4 status was examined via 5'-exonuclease assay, detecting 6 calves carrying the defect. These were purchased and monitored daily until final necropsy. The calves developed progressive skin lesions starting with small scaling areas terminating in severe crusting dermatitis, especially in areas with mechanical exposure. Histological and immunohistochemical analyses indicated that macrophages with cytoplasmic vacuolation increased considerably in skin sections obtained weekly during the disease course. Macrophage increase correlated with increased dermal lesion severity. Macrophage activation was confirmed by prominent phagocytic activity in the superficial dermis using electron microscopy. Dermal mRNA abundance of CCL2 and CCL3 measured by quantitative polymerase chain reaction verified macrophage activation. Further increase in mRNA of downstream molecule MyD88 and cytokine IL12b connected bovine PLD4 deficiency to increased TLR9 pathway activation. In contrast to human macrophage activation syndrome, the main feature of bovine PLD4 deficiency was local disease in organs with contact to microbial DNA (skin, intestine, lungs).

A High Plane of Nutrition Is Associated with a Lower Risk for Neonatal Calf Diarrhea on Bavarian Dairy Farms.

In all bovine production systems, neonatal calf diarrhea remains worldwide an important issue of economic losses and animal welfare. The aim of the present study was to identify risk factors for neonatal calf diarrhea as a herd health problem on Bavarian dairy farms. For the purpose of this study, management factors related to calf health were retrospectively compared between 59 dairy farms with calf diarrhea as a herd problem with those of 18 control farms, where no veterinary treatment of calves for neonatal calf diarrhea took place for at least one year prior to the farm visit. A multivariable binary logistic regression analysis of management factors indicated that administration of 3 L or more of colostrum at the second feeding after birth (Odds ration [OR] = 0.21, 95% confidence interval [95% CI] = 0.05-0.89), ad libitum feeding of milk during the first week of life (OR = 0.06, 95% CI = 0.006-0.60), and administration of an iron containing preparation after birth (OR = 10.9, 95% CI = 1.25-95.6) were independently associated with the presence of a herd problem with neonatal diarrhea. Results of this study therefore suggest that a higher plane of nutrition is a protective factor with regard to the occurrence of neonatal diarrhea on Bavarian dairy farms. These findings support the establishment of ad libitum feeding programs in dairy calf rearing.

Dependence of the apparent bicarbonate space on initial plasma bicarbonate concentration and carbon dioxide tension in neonatal calves with diarrhea, acidemia, and metabolic acidosis.

BACKGROUND: Marked strong ion (metabolic) acidosis in neonatal diarrheic calves usually is corrected by IV administration of NaHCO3 . The distribution space for IV-administered bicarbonate, called the apparent bicarbonate space (ABS), appears to depend on initial plasma bicarbonate concentration (cHCO3 ) and varies considerably in calves. OBJECTIVE: To determine whether ABS was associated with initial plasma cHCO3 and other acid-base variables. ANIMALS: Twenty-five neonatal diarrheic calves with acidemia and metabolic acidosis. METHODS: Prospective observational study using a convenience sample. Calves received NaHCO3 (10 mmol/kg) and glucose (1.4 mmol/kg) IV in a crystalloid solution at 25 mL/kg over 60 minutes. The ABS (L/kg) was calculated at 4 time points over 2 hours after the end of the infusion. The relationship between ABS and initial acid-base variables was characterized using nonlinear, linear, and stepwise regression. RESULTS: The median value for ABS calculated from the initial plasma cHCO3 increased from 0.53 L/kg (range, 0.40-0.79) at the end of IV infusion to 0.96 L/kg (range, 0.54-1.23) 120 minutes later. Data obtained at the end of infusion provided the best fit to initial plasma cHCO3 and jugular venous blood Pco2 , such that: ABS = 0.41 + 1.06/cHCO3 and ABS = 0.87-0.0082 × Pco2 . CONCLUSIONS AND CLINICAL IMPORTANCE: The observed median value for ABS of 0.53 L/kg in our study was similar to the empirically used value of 0.6. However, ABS values varied widely and were increased in calves with severe metabolic acidosis. We therefore recommend calculating ABS using the initial plasma cHCO3 or venous blood Pco2 , if respective measurements are available.

Intravenous and Oral Fluid Therapy in Neonatal Calves With Diarrhea or Sepsis and in Adult Cattle.

Optimal fluid therapy protocols in neonatal calves and adult cattle are based on consideration of signalment, history, and physical examination findings, and individually tailored whenever laboratory analysis is available. Measurement of the magnitude of eye recession, duration of skin tenting in the lateral neck region, and urine specific gravity by refractometry provide the best estimates of hydration status in calves and cattle. Intravenous and oral electrolyte solutions (OES) are frequently administered to critically ill calves and adult cattle. Application of physicochemical principles indicates that 0.9% NaCl, Ringer's solution, and 5% dextrose are equally acidifying, lactated Ringer's and acetated Ringer's solution are neutral to mildly acidifying, and 1.3-1.4% sodium bicarbonate solutions are strongly alkalinizing in cattle. Four different crystalloid solutions are recommended for intravenous fluid therapy in dehydrated or septic calves and dehydrated adult cattle: (1) lactated Ringer's solution and acetated Ringer's solution for dehydrated calves, although neither solution is optimized for administration to neonatal calves or adult cattle; (2) isotonic (1.3%) or hypertonic (5.0 or 8.4%) solutions of sodium bicarbonate for the treatment of calves with diarrhea and severe strong ion (metabolic) acidosis and hyponatremia, and adult cattle with acute ruminal acidosis; (3) Ringer's solution for the treatment of metabolic alkalosis in dehydrated adult cattle, particularly lactating dairy cattle; and (4) hypertonic NaCl solutions (7.2%) and an oral electrolyte solution or water load for the rapid resuscitation of dehydrated neonatal calves and adult cattle. Much progress has been made since the 1970's in identifying important attributes of an OES for diarrheic calves. Important components of an OES for neonatal calves are osmolality, sodium concentration, the effective SID that reflects the concentration of alkalinizing agents, and the energy content. The last three factors are intimately tied to the OES osmolality and the abomasal emptying rate, and therefore the rate of sodium delivery to the small intestine and ultimately the rate of resuscitation. An important need in fluid and electrolyte therapy for adult ruminants is formulation of a practical, effective, and inexpensive OES.

Effects of pH and the plasma or serum concentrations of total calcium, chloride, magnesium, l-lactate, and albumin on the plasma ionized calcium concentration in calves.

BACKGROUND: The plasma ionized calcium concentration (cCa2+ ) represents the biologically active form of calcium and is the preferred method for evaluating calcium status in animals. Different pH-corrective equations have been developed for human plasma, but the validity of the equations for bovine plasma is unknown. HYPOTHESIS: We hypothesized that pH-corrective equations for bovine plasma would be similar to those used for human plasma; cCa2+ was dependent on the plasma concentrations of total calcium (cTCa), chloride (cCl), L-lactate (cLactate), and albumin (cAlbumin); and the in vitro and in vivo cCa2+ -pH relationships would differ. ANIMALS: Ten healthy calves (in vitro study), 1426 critically ill calves. METHODS: The in vitro plasma log10 (cCa2+ )-pH relationship was determined by CO2 tonometry of 465 plasma samples. Plasma cCl was altered by equivolume dilution of plasma with 3 electrolyte solutions of different cCl. The in vivo plasma cCa2+ -pH relationship was investigated and validated using clinicopathologic data extracted from the medical records of 950 (model development) and 476 (model validation) critically ill calves. RESULTS: pH-corrective equations for bovine plasma were similar to those used for human plasma. Plasma cCa2+ increased in vitro with increases in plasma cCl. Plasma cCa2+ in critically ill calves was associated with plasma cTCa, blood pH, plasma cCl, serum cMg, and cL-lactate (R2 = 0.69) but not plasma cAlbumin. CONCLUSIONS AND CLINICAL IMPORTANCE: Calculation of cCa2+ from cTCa in calf plasma or serum requires adjustment for at least pH and cCl when 1 or both are outside the reference range.

Electrocardiographic findings in 130 hospitalized neonatal calves with diarrhea and associated potassium balance disorders.

BACKGROUND: Hyperkalemia in neonatal diarrheic calves can potentially result in serious cardiac conduction abnormalities and arrhythmias. OBJECTIVES: To document electrocardiographic (ECG) findings and the sequence of ECG changes that are associated with increasing plasma potassium concentrations (cK+ ) in a large population of neonatal diarrheic calves. ANIMALS: One hundred and thirty neonatal diarrheic calves (age ≤21 days). METHODS: Prospective observational study involving calves admitted to a veterinary teaching hospital. RESULTS: Hyperkalemic calves (cK+ : 5.8-10.2, blood pH: 6.55-7.47) had significantly (P < .05) longer QRS durations as well as deeper S wave, higher T wave, and higher ST segment amplitudes in lead II than calves, which had both venous blood pH and cK+ within the reference range. The first ECG changes in response to an increase in cK+ were an increase in voltages of P, Ta, S, and T wave amplitudes. Segmented linear regression indicated that P wave amplitude decreased when cK+ >6.5 mmol/L, S wave amplitude voltage decreased when cK+ >7.4 mmol/L, QRS duration increased when cK+ >7.8 mmol/L, J point amplitude increased when cK+ >7.9 mmol/L, and ST segment angle increased when cK+ >9.1 mmol/L. P wave amplitude was characterized by a second common break point at cK+ = 8.2 mmol/L, above which value the amplitude was 0. CONCLUSIONS AND CLINICAL IMPORTANCE: Hyperkalemia in neonatal diarrheic calves is associated with serious cardiac conduction abnormalities. In addition to increased S and T wave amplitude voltages, alterations of P and Ta wave amplitudes are early signs of hyperkalemia, which is consistent with the known sensitivity of atrial myocytes to increased cK+ .

Plasma potassium concentrations in neonatal diarrhoeic calves are correlated with serum aldosterone concentrations but not with insulin concentrations.

Hyperkalaemia is a clinically relevant electrolyte imbalance in neonatal diarrhoeic calves which was previously associated with severe dehydration and acidaemia. The present study assessed the association of plasma potassium (cK) with serum aldosterone and insulin concentrations, since these hormones are involved in the regulation of potassium homeostasis. Serum aldosterone (rs=0.62), but not insulin concentrations (rs=0.22) were closely correlated to cK in 123 hospitalised neonatal diarrhoeic calves. Median values for serum aldosterone concentrations in 38 hyperkalaemic calves (cK>5.8mmol/L) were 3.2 and 15.3 times higher (P<0.001) than in 85 non-hyperkalaemic diarrhoeic calves and nine healthy control calves, respectively. Aldosterone, but not insulin secretion, appears to be highly stimulated in dehydrated diarrhoeic calves with hyperkalaemia, but hypovolaemia and a concomitant decrease in renal perfusion and urinary flow rate are likely to limit the efficacy of aldosterone-induced control mechanisms in these animals.

Clinical signs, profound acidemia, hypoglycemia, and hypernatremia are predictive of mortality in 1,400 critically ill neonatal calves with diarrhea.

Profound acidemia impairs cellular and organ function and consequently should be associated with an increased risk of mortality in critically ill humans and animals. Neonatal diarrhea in calves can result in potentially serious metabolic derangements including profound acidemia due to strong ion (metabolic) acidosis, hyper-D-lactatemia, hyper-L-lactatemia, azotemia, hypoglycemia, hyperkalemia and hyponatremia. The aim of this retrospective study was to assess the prognostic relevance of clinical and laboratory findings in 1,400 critically ill neonatal calves with diarrhea admitted to a veterinary teaching hospital. The mortality rate was 22%. Classification tree analysis indicated that mortality was associated with clinical signs of neurologic disease, abdominal emergencies, cachexia, orthopedic problems such as septic arthritis, and profound acidemia (jugular venous blood pH < 6.85). When exclusively considering laboratory parameters, classification tree analysis identified plasma glucose concentrations < 3.2 mmol/L, plasma sodium concentrations ≥ 151 mmol/L, serum GGT activity < 31 U/L and a thrombocyte count < 535 G/L as predictors of mortality. However, multivariable logistic regression models based on these laboratory parameters did not have a sufficiently high enough sensitivity (59%) and specificity (79%) to reliably predict treatment outcome. The sensitivity and specificity of jugular venous blood pH < 6.85 were 11% and 97%, respectively, for predicting non-survival in this study population. We conclude that laboratory values (except jugular venous blood pH < 6.85) are of limited value for predicting outcome in critically ill neonatal calves with diarrhea. In contrast, the presence of specific clinical abnormalities provides valuable prognostic information.

The prevalence and clinical relevance of hyperkalaemia in calves with neonatal diarrhoea.

One hundred and twenty-four calves with neonatal diarrhoea were investigated in order to assess the prevalence of hyperkalaemia and the associated clinical signs. Hyperkalaemia (potassium concentration >5.8mmol/L) was recognized in 42 (34%) calves and was more closely associated with dehydration than with decreases in base excess or venous blood pH. In 75 calves with normal blood concentrations of D-lactate (i.e. ⩽3.96mmol/L), K concentrations were moderately correlated with base excess values (r=-0.48, P<0.001). In contrast, no significant correlation was observed in 49 calves with elevated D-lactate. Only three hyperkalaemic calves had bradycardia and a weak positive correlation was found between heart rate and K concentrations (r=0.22, P=0.014). Ten of the 124 calves had cardiac arrhythmia and of these seven had hyperkalaemia indicating that cardiac arrhythmia had a low sensitivity (17%) but a high specificity (96%) as a predictor of hyperkalaemia. In a subset of 34 calves with base excess values ⩽-5mmol/L and D-lactate concentrations <5mmol/L (of which 22 had hyperkalaemia), changes in posture/ability to stand could be mainly explained by elevations of K concentrations (P<0.001) and to a lesser extent by increases in L-lactate concentrations (P=0.024). Skeletal muscle weakness due to hyperkalaemia alongside hypovolaemia may produce a clinical picture that is similar to that in calves with marked D-lactic acidosis. However, since reductions in the strength of the palpebral reflex are closely correlated with D-lactate concentrations, a prompt palpebral reflex can assist the clinical prediction of hyperkalaemia in calves presenting with a distinct impairment in their ability to stand (specificity 99%, sensitivity 29%).

Construction and validation of a decision tree for treating metabolic acidosis in calves with neonatal diarrhea.

BACKGROUND: The aim of the present prospective study was to investigate whether a decision tree based on basic clinical signs could be used to determine the treatment of metabolic acidosis in calves successfully without expensive laboratory equipment. A total of 121 calves with a diagnosis of neonatal diarrhea admitted to a veterinary teaching hospital were included in the study. The dosages of sodium bicarbonate administered followed simple guidelines based on the results of a previous retrospective analysis. Calves that were neither dehydrated nor assumed to be acidemic received an oral electrolyte solution. In cases in which intravenous correction of acidosis and/or dehydration was deemed necessary, the provided amount of sodium bicarbonate ranged from 250 to 750 mmol (depending on alterations in posture) and infusion volumes from 1 to 6.25 liters (depending on the degree of dehydration). Individual body weights of calves were disregarded. During the 24 hour study period the investigator was blinded to all laboratory findings. RESULTS: After being lifted, many calves were able to stand despite base excess levels below -20 mmol/l. Especially in those calves, metabolic acidosis was undercorrected with the provided amount of 500 mmol sodium bicarbonate, which was intended for calves standing insecurely. In 13 calves metabolic acidosis was not treated successfully as defined by an expected treatment failure or a measured base excess value below -5 mmol/l. By contrast, 24 hours after the initiation of therapy, a metabolic alkalosis was present in 55 calves (base excess levels above +5 mmol/l). However, the clinical status was not affected significantly by the metabolic alkalosis. CONCLUSIONS: Assuming re-evaluation of the calf after 24 hours, the tested decision tree can be recommended for the use in field practice with minor modifications. Calves that stand insecurely and are not able to correct their position if pushed require higher doses of sodium bicarbonate, if there is clinical evidence of a marked D-lactic acidosis. In those calves, determining the degree of loss of the palpebral reflex was identified as a useful decision criterion to provide an additional amount of 250 mmol sodium bicarbonate. This work demonstrates the clinical relevance of the discovery that D-lactate is responsible for most of the clinical signs expressed in neonatal diarrheic calves suffering from metabolic acidosis.