Acute phase protein levels in pregnancy and oestrus cycle in bitches

The purpose of this study was to evaluate the acute phase protein (serum amyloid A, C-reactive protein, haptoglobin, ceruloplasmin and fibrinogen) profiles in pregnant dogs as well as bitches at various stages of the estrus cycle and to verify possible relationship(s) among the APPs profile and hormonal status. A total of 60 bitches of various breeds were enrolled. The animals were divided into six groups. B-mode ultrasonographic scanner (concept M/C) was used for pregnancy diagnosis, involving the first and second halves of pregnancy. Proestrus, estrus, diestrus and anestrus stages of sexual cycles were differentiated by use of vaginal cytology and blood progesterone and estradiol 17-β levels. Serum C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin, ceruloplasmin and fibrinogen levels were determined. No statistically significant differences were observed between the groups with regard to the serum SAA and CRP concentrations. Haptoglobin, ceruloplasmin and fibrinogen concentrations were elevated in pregnant dogs compared to non-pregnant groups’, independently from the pregnancy stage. No statistically significant differences in acute phase protein concentrations in dogs at various stages of estrus could be detected. Results of this study indicated that ceruloplasmin and fibrinogen levels may have potential importance in the diagnosis of pregnancy, and acute phase proteins may be used for monitoring the health status of the pregnant dogs.     

Acute phase protein response in dogs with experimentally induced gastric mucosal injury

Background: The acute phase response is part of the innate defense system of an animal against trauma, inflammation, and infection. Objective: The purpose of this investigation was to evaluate the acute phase response in dogs with acetylsalicylic acid-induced gastric mucosal injuries and to determine its potential diagnostic significance. Methods: Ten, healthy, cross-breed dogs (6 females and 4 males) were given oral acetylsalicylic acid in a single oral dose of 200 mg/kg for the experimental model of acute gastric mucosal injury. Heparinized blood samples were collected before (day 0) and on days 1, 4, and 7 after acetylsalicylic acid administration to determine plasma C-reactive protein (CRP), serum amyloid-A (SAA), haptoglobin, fibrinogen, total protein, albumin, and iron concentrations. Total WBC counts were done in whole blood. Endoscopy was done on each of the selected days, and gastric lesions were scored and localized. Results: Hemorrhagic linear erosions were found in all dogs on day 1, concurrent with significant increases in CRP, SAA, haptoglobin, and fibrinogen concentrations, and in WBC counts; however, iron concentration was decreased. Peak haptoglobin and fibrinogen concentrations occurred on day 4, at which time only nonhemorrhagic lesions were observed endoscopically. On day 7, results for all analytes except haptoglobin had returned to baseline levels, along with resolution of most gastric lesions. Conclusions: Results of this study indicate that a rapid acute phase protein response occurs after induced gastric mucosal injury in dogs and may be potentially useful together with gastroscopy in the diagnosis and monitoring of gastric injury.     

Serum acute phase protein concentrations in dogs with hyperadrenocorticism with and without concurrent inflammatory conditions

Background: Acute phase proteins (APPs) are promising markers of inflammation in dogs, because they are more sensitive than WBC counts in detecting clinical and subclinical inflammation. Endogenous corticosteroids can mask an acute phase response and make it more difficult to identify underlying inflammatory disease. Objective: The purpose of this study was to evaluate the acute phase protein response in dogs with spontaneous hyperadrenocorticism (HAC) with and without concurrent inflammatory conditions. Methods: Serum concentrations of C‐reactive protein (CRP), haptoglobin (Hp), fibrinogen, and albumin were measured in 44 healthy adult dogs and 39 dogs with HAC; the HAC group was further divided into dogs with and without concurrent infection/inflammation. A fourth group of dogs with severe sepsis and without HAC was compared with the dogs with HAC and severe sepsis. Results: Dogs with uncomplicated HAC had significantly higher Hp and fibrinogen concentrations compared with healthy control dogs (P<.001). Dogs with HAC and severe inflammatory disease also had significantly higher CRP and lower albumin concentrations than control dogs and dogs with HAC without concurrent inflammation. Dogs with sepsis but without HAC had significantly higher CRP concentrations than dogs with HAC and sepsis. Conclusions: Dogs with HAC had increases in the moderate APPs (Hp and fibrinogen), and no significant changes in CRP and albumin compared with healthy dogs. Although concurrent HAC appeared to blunt the CRP response in dogs with sepsis, increased serum CRP concentration in dogs with HAC is likely indicative of severe concurrent inflammation.     

Reference ranges and the influence of age and sex on hematologic and serum biochemical values in donkeys (Equus asinus)

Hematologic and serum biochemical values were determined in blood samples from 217 donkeys (Equus asinus). Donkeys were classified on the basis of size, sex, age, and whether they were domestic or feral. Parametric (mean +/- 2 SD) and nonparametric (2.5th to 97.5th percentile) reference ranges were calculated for each analyte. For all donkeys, 26 of 46 analytes significantly departed from gaussian distribution. Serum lactate dehydrogenase activity in miniature donkeys was higher than that in other donkeys. Differential leukocyte counts in feral donkeys differed from those in other types in ways that suggested that the former had smaller parasite loads or experienced greater stress. Erythrocyte, lymphocyte, and platelet counts and fibrinogen, glucose, inorganic phosphorus, and potassium concentrations decreased with age. Eosinophil counts, mean corpuscular volume, and plasma protein, serum protein, and serum globulin concentrations increased with age. Female donkeys had significantly (P less than 0.05) higher mean corpuscular hemoglobin concentration and leukocyte and neutrophil counts than did male donkeys. Mean corpuscular hemoglobin increased with age, and females had higher values than did males of all age groups. An interaction between age and sex was observed for alkaline phosphatase activity, with a trend for decreased activity with age.     

Prognostic value of serum acute-phase proteins in dogs with parvoviral enteritis

Objective : To evaluate the acute-phase protein response in dogs with parvoviral enteritis as predictor of the clinical outcome. Methods : Canine parvovirus infection was diagnosed based on the compatible clinical findings and confirmed by the canine parvovirus antigen test in 43 dogs of less than six months of age. Blood samples for complete blood cell count and acute-phase proteins (C-reactive protein, haptoglobin, ceruloplasmin and albumin) were collected before treatment. Twenty-three dogs died during or after treatment (non-survival) and the rest recovered (survival). Five healthy dogs were enrolled as control. Results : Serum C-reactive protein, ceruloplasmin and haptoglobin levels in dogs with parvoviral enteritis were higher (P<0·001, P<0·01 and P<0·001, respectively), but serum albumin was lower (P<0·001) than those in controls. Mean C-reactive protein and ceruloplasmin values in non-survival were higher (P<0·01) than those for survival dogs. C-reactive protein was found to be superior to ceruloplasmin, haptoglobin and albumin for distinguishing survival from non-survival dogs. Values higher than 92·4 mg/l for C-reactive protein had a sensitivity of 91% to predict mortality. Clinical Significance : The magnitude of the increase in serum acute-phase proteins in dogs with parvoviral enteritis could be a useful indicator of the prognosis of the disease. In acute-phase proteins, C-reactive protein is a potent predictor of mortality in dogs with parvoviral enteritis.     

Bovine and canine acute phase proteins

Acute phase proteins are serum proteins which increase in concentration during the acute phase response to inflammation or infection. The response occurs in all animals, but in different species the response of individual proteins can be significantly different. Of the numerous acute phase proteins which have been identified in humans, a number have been examined in cattle and dogs but usually on an individual basis with little reference to their part in the acute phase response. Biochemical, physiological and clinical investigations into haptoglobin, fibrinogen, ₁-proteinase inhibitor, ceruloplasmin, seromucoid and C-reactive protein of cattle and dogs have therefore been reviewed with the emphasis on their role in this response to tissue damage.     

The effect of copper deficiency on the peripheral blood cells of cattle

Six female cattle were given molybdenum (30 ppm) and sulphate (225 ppm) to induce experimental secondary copper deficiency. The total and differential leukocyte numbers and lymphocyte subpopulations were counted and the neutrophil activity was assessed by means of nitroblue tetrazolium reduction and phagocytosis of sheep erythrocytes. The serum caeruloplasmin activity and concentration were also determined. Copper deficiency was confirmed from decreased serum copper levels, the animals with values less than 5.9 µmol/L being considered deficient. Total leukocyte numbers were not affected by the copper deficiency. However, differential counts showed a marked increase in monocyte subpopulations, a significant decrease in B lymphocytes and reduced neutrophil activity. The serum caeruloplasmin activity was decreased about 50%, but the total serum protein concentration was less altered. We concluded that the effect of these changes on the animals' immune competence may contribute to a greater incidence of infectious diseases in copper-deficient cattle.     

Serum concentration of some acute phase proteins in naturally occurring canine babesiosis: a preliminary study

BACKGROUND: Serum concentrations of acute phase proteins can provide valuable diagnostic information in the detection, prognosis, or monitoring of disease. Information available on the acute phase response in naturally occurring canine babesiosis is limited. OBJECTIVE: The purpose of this investigation was to retrospectively evaluate serum concentrations of haptoglobin, C-reactive protein, and ceruloplasmin in dogs naturally infected with Babesia canis. METHODS: Haptoglobin, C-reactive protein, and ceruloplasmin concentrations were measured in serum samples from dogs with uncomplicated (n = 6) and complicated (n = 1) babesiosis and compared with 6 healthy dogs. RESULTS: Serum C-reactive protein and ceruloplasmin concentrations were significantly higher in dogs with babesiosis; however, serum haptoglobin concentration was significantly lower compared with control dogs (P <.01). CONCLUSIONS: Results of this study suggest that acute phase protein concentrations could be beneficial in the diagnosis and determination of the severity of babesiosis in dogs.     

Serum concentrations of acute-phase proteins in dogs with leishmaniosis during short-term treatment

OBJECTIVE: To evaluate changes in serum concentrations of acute-phase proteins in dogs with leishmaniosis during short-term therapy in accordance with 2 treatment protocols and determine whether concentrations of acute-phase proteins could be used to monitor the initial response of dogs to treatment. ANIMALS: 12 dogs naturally infected with Leishmania infantum. PROCEDURE: Dogs were allocated into 2 groups. Dogs of group 1 were treated by use of meglumine antimonate (100 mg/kg, SC, q 24 h) administered concurrently with allopurinol (15 mg/kg, PO, q 12 h) for 20 days and then with allopurinol alone at the same dosage for the subsequent 30 days. Dogs of group 2 were treated by administration of allopurinol alone (15 mg/kg, PO, q 12 h) for 60 days). Blood samples were obtained before and during treatment for measurement of serum concentrations of acute-phase proteins and determination of CBC counts, serum biochemical analyses, and electropherograms. RESULTS: All dogs evaluated in the study had increased concentrations of C-reactive protein, haptoglobin, and ceruloplasmin at the time of diagnosis of leishmaniosis. Mean concentration of serum amyloid A before treatment was also increased, but some of the dogs had concentrations of serum amyloid A that were within the reference range. Concentrations of C-reactive protein and ceruloplasmin decreased significantly in all dogs at the end of the study period. CONCLUSIONS AND CLINICAL RELEVANCE: Measurement of concentrations of selected acute-phase proteins, such as C-reactive protein or ceruloplasmin, could be used to evaluate the initial response of dogs with leishmaniosis to treatment.     

Acute phase proteins in grass sickness (equine dysautonomia)

Four acute phase proteins were assayed in the serum of normal horses and those with acute, subacute and chronic grass sickness, colic and inflammatory conditions, in order to investigate their diagnostic value in grass sickness. The grass sickness and inflammation group had a significantly increased haptoglobin concentration (P less than 0.01-P less than 0.001). Orosomucoid was elevated in acute, subacute and chronic grass sickness and inflammation (P less than 0.001, P less than 0.001, P less than 0.05 and P less than 0.05, respectively). Highest concentrations of haptoglobin and orosomucoid were recorded in subacute grass sickness. Ceruloplasmin was significantly higher in acute grass sickness cases than all other groups except the colic group (P less than 0.05-P less than 0.01). alpha 2-macroglobulin was significantly higher in acute grass sickness than normal, colic and chronic grass sickness cases (P less than 0.01, P less than 0.05 and P less than 0.05). The time scale of changes suggests that the stimulus to haptoglobin and orosomucoid synthesis occurs at the onset of clinical signs whereas the increase in ceruloplasmin and alpha 2-macroglobulin is more likely to reflect haemoconcentration.     

Acute phase response of sheep: changes in the concentrations of ceruloplasmin, fibrinogen, haptoglobin and the major blood cell types associated with pulmonary damage

The concentrations of plasma ceruloplasmin, plasma fibrinogen, serum haptoglobin and the major cell types in blood together with liveweight changes were monitored during the acute phase response in sheep. Five control sheep, five sheep that underwent sham bronchial obstruction, and five sheep that developed pneumonia after bronchial obstruction were examined. Blood samples were taken and liveweights were recorded from four to six days before until 14 days after the surgical operations (sham and bronchial obstruction). The operations led to an acute phase response in the sheep and the development of pneumonia increased and sustained the response or led to a secondary response. Statistically significant changes observed in the blood of the sheep during the acute phase response included increased concentrations of plasma ceruloplasmin and plasma fibrinogen, depression of erythrocyte numbers and elevation of neutrophil numbers (means on day of maximum change as percentage of pretreatment values; 250 per cent, 400 per cent, 80 per cent and 200 per cent, respectively). Serum haptoglobin showed a pronounced and significant increase in concentration (over 6000 per cent of pretreatment values in some sheep). All three groups of sheep showed significant depression of liveweight after overnight confinement in the surgery but this was sustained for the period of the experiment only in the bronchial obstruction group. The results indicated that measurement of the concentrations of the three plasma proteins may be more useful in the diagnosis of tissue injury and infectious disease than the number of circulating neutrophils in sheep.     

Ceruloplasmin as an indicator of copper status in cattle and sheep.

The relationship between ceruloplasmin, a metalloenzyme with oxidase activity, and copper was investigated in cattle and sheep. The oxidase activity of ceruloplasmin correlated closely with the serum or plasma copper concentrations in cattle. The respective correlation coefficients were 0.83 and 0.60. In sheep serum, a correlation coefficient of 0.92 was obtained. In each instance, the relationship remained linear from the deficient to the high normal ranges of copper. Comparison of the linear regression relationships indicated the ceruloplasmin activity in bovine serum was statistically lower than the activity in bovine plasma (P less than 0.0001), through the intercepts from the regression lines of the two relationships were similar (P = 0.412). Comparisons of ovine and bovine serum-ceruloplasmin relationships indicated that a significant species difference was present. Ovine ceruloplasmin activity increased more rapidly as compared to the corresponding bovine activity over the range of copper concentrations investigated (P less than 0.0001). The intercept from the ovine regression relationship was also lower (P less than 0.0001). A correlation coefficient of 0.35 was observed between the serum ceruloplasmin activity and hepatic copper concentrations in cattle indicating that the mathematical relationship was not as well defined. Ceruloplasmin activity appears to correlate more closely with serum or plasma copper concentrations as compared to corresponding liver copper concentrations.     

Acute phase response in the dog following surgical trauma

The levels of five acute phase reactants and serum glycoproteins were measured in dogs following surgical trauma. Canine C-reactive protein peaked 24 hours after surgery and peak levels of ceruloplasmin, haptoglobin and seromucoid were detected on the fourth to sixth days. Levels of serum glycoproteins varied in a similar manner to ceruloplasmin, haptoglobin and seromucoid. There was very little variation in the levels of alpha 1 antitrypsin.     

Bovine acute phase response following turpentine injection

The serum levels of five proteins, alpha 1 antitrypsin, ceruloplasmin, fibrinogen, haptoglobin and seromucoid, were measured daily in calves after the subcutaneous injection of oil of turpentine. Raised concentrations were detected on the second and third days after injection with peak levels occurring on the fourth to seventh days and returning to normal by the 17th day. Levels of four of these proteins, alpha 1 antitrypsin, ceruloplasmin, haptoglobin and seromucoid were compared in the same calves following three different doses of turpentine. Levels of haptoglobin and seromucoid varied with the dose whereas ceruloplasmin and alpha 1 antitrypsin levels did not.     

Acute phase proteins as indicators of calf herd health

The potential for using acute phase proteins (APPs) in the assessment of herd health was studied by examining the levels of serum haptoglobin, serum amyloid A (SAA) and plasma fibrinogen in relation to clinical findings and leukocyte counts in calves. Two groups of calves from conventional dairy farms were studied. The animals were examined 10 times during the first six weeks after introduction into a new environment. Haptoglobin, SAA and fibrinogen were analysed and weight gain, disease symptoms and treatments were recorded. Analysis of antibodies against viral infections was performed. An acute phase reaction (APR) score was established at each sampling by combining the APP results and total leukocyte counts. The health status differed between the two groups, although no manipulation of health had been performed, except that the group with a higher incidence of disease had a concurrent experimental infection with lungworm as part of another study. In the group with a higher incidence of disease, the mean weight gain was significantly lower, and the number of sampling days with elevated serum concentrations of APPs, and the mean maximum concentrations of haptoglobin and fibrinogen were significantly higher compared to the healthier group. The APR score was significantly higher at days 4 and 8 of the study in the group with a higher incidence of disease. The results indicate that measurement of APPs could be a useful tool for evaluation of health in calf herds.     

Effects of short-term trimethoprim-sulfamethoxazole administration on thyroid function in dogs

OBJECTIVE: To determine how rapidly trimethoprim-sulfamethoxazole affects serum total thyroxine (T4) and thyroid-stimulating hormone (TSH) concentrations in euthyroid dogs and how quickly hormone concentrations return to reference values following discontinuation of administration. DESIGN: Prospective study. ANIMALS: 7 healthy euthyroid dogs. PROCEDURE: Dogs were given trimethoprim-sulfamethoxazole (26.5 to 31.3 mg/kg [12 to 14.2 mg/lb], PO, q 12 h) for a maximum of 6 weeks. A CBC and Schirmer tear test were performed and serum total T4 and TSH concentrations were measured weekly. Administration of trimethoprim-sulfamethoxazole was discontinued if total T4 concentration was less than the lower reference limit and TSH concentration was greater than the upper reference limit or if persistent neutropenia developed. RESULTS: Six dogs had total T4 concentrations less than the lower reference limit within 3 weeks; T4 concentration was decreased after 1 week in 3 of these 6 dogs. In these 6 dogs, TSH concentration was greater than the upper reference limit within 4 weeks. In 1 dog, T4 and TSH concentrations were not affected, despite administration of trimethoprim-sulfamethoxazole for 6 weeks. Neutropenia developed in 4 dogs. In 1 dog, the neutropenia resolved while trimethoprim-sulfamethoxazole was still being administered. In the other 3, neutrophil counts returned to reference values 1 week after drug administration was discontinued. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of trimethoprim-sulfamethoxazole at a dosage of 26.5 to 31.3 mg/kg, PO, every 12 hours can substantially alter serum total T4 and TSH concentrations and neutrophil counts in dogs within as short a time as a few weeks.     

Acute phase response in horses: changes in plasma cation concentrations after localised tissue injury

An acute phase reaction was elicited in four horses to which Freund's adjuvant was administered intramuscularly. The localised inflammation was accompanied by changes in the plasma concentrations of copper, iron and zinc. The plasma copper concentration, the plasma ceruloplasmin copper concentration and the ceruloplasmin oxidase activity in the plasma steadily increased to a maximum 24 days after the administration of the adjuvant. At this time, the plasma copper concentration was 2.2 micrograms/ml, a 90 per cent increase over the baseline concentration. The ratio of the concentration of plasma ceruloplasmin copper to plasma copper remained constant, indicating that the non-ceruloplasmin bound copper component of the plasma is also an acute phase reactant in the horse. The plasma zinc and iron concentrations decreased to 59 per cent and 30 per cent of their respective baseline concentrations and the severity of the inflammation appeared to influence the plasma concentrations of each metal. Weak correlations between the plasma fibrinogen concentration and the plasma copper and zinc concentrations of 25 horses with plasma fibrinogen concentrations of 5 g/litre or greater indicated that a single measurement of plasma copper concentration is not useful in the diagnosis of non-specific inflammatory disorders of the horse. However, the results suggest that the plasma copper concentrations in serial samples may be used to monitor the resolution of inflammatory disorders in the horse.     

Leukocyte and platelet changes following low-dose lipopolysaccharide administration in five dogs

Effects of low-dose LPS (0.1 μg/kg IV) on leukocyte and platelet parameters measured using an Advia 120 hematology analyzer were investigated. Five dogs received a saline sham treatment prior to LPS, and blood was collected before and 3, 6, and 24 h post-treatment. LPS-treated dogs had mild neutrophil toxic change and increased neutrophil bands at 3 and 6 h. Compared to saline-treated controls, total leukocyte, neutrophil, and monocyte counts of LPS-treated dogs were significantly decreased at 3 h and increased at 24 h. Compared to baseline, total leukocyte counts of LPS-treated dogs were significantly decreased at 3 h and increased at 24 h. Mean platelet volume was significantly increased and mean platelet component concentration was decreased at 3 h compared to baseline. Platelet count was significantly decreased at 3 and 6 h; plateletcrit did not change significantly. High dosage is not required in order to detect LPS-mediated hematologic effects in dogs. Low-dose LPS administration causes significant changes in leukocyte and platelet indices in dogs without causing severe clinical signs or death.     

An automated spectrophotometric method for measuring canine ceruloplasmin in serum

An automated method for the determination of ceruloplasmin activity was developed and validated in canine serum. The method is based on the in vitro oxidase activity that this protein shows with substances such as p-phenylenediamine. In order to determine optimum assay conditions, the effects of the substrate concentration, buffer pH, reaction time and EDTA on the reaction were evaluated. The precision of the assay was good with within-run and between-run coefficients of variation lower than 10%. The method measured the ceruloplasmin values in a proportional and linear manner (r = 0.99) with a limit of detection of 0.0007 +/- 0.0001 Delta Abs/min. A temperature of -20 degrees C kept the reagent stable for 30 days. The method is cheap and easy to adapt to any automated biochemical analyser, considerably decreasing the processing time required with the manual method. Additionally it allows to differentiate dogs with pyometra and trauma from clinically healthy dogs.     

Serum concentrations of acute phase proteins in dogs with leishmaniasis

The concentrations of haptoglobin, C-reactive protein and ceruloplasmin were measured in symptomatic and asymptomatic dogs naturally infected by Leishmania infantum, and in healthy uninfected dogs to determine the potential value of these proteins for the diagnosis and prognosis of leishmaniasis. The concentrations of the acute phase proteins were significantly higher in the dogs with leishmaniasis than in the control dogs, and the concentration of C-reactive protein was significantly higher in the symptomatic dogs than in the asymptomatic dogs. There were no correlations between the acute phase proteins and the gamma globulins, the albumin/globulin ratio or the titre of anti-leishmanial antibodies.