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.     

Anaemia in housed lambs

Anaemia was detected in housed lambs by clinical and haematological investigation. Conjunctival pallor was used as a clinical test for anaemia and the results indicate that this has high specificity (91 per cent to 95 per cent) and low sensitivity (53 per cent to 55 per cent). The haematological results indicated a non-regenerative anaemia with low packed cell volume, red blood cell count and haemoglobin. In a subset of lambs examined biochemically, anaemia was associated with low serum iron concentration and low serum iron binding: cobalt levels were within normal ranges and blood copper levels were slightly raised. At present it is unclear whether this is a primary or secondary iron deficiency.     

A Simple Method for Separation of Buffy Coat from Peripheral Blood of Chickens

A simple pasteur pipette tube technique is described for the separation of buffy coat from the peripheral blood of chickens. Whole blood treated with disodium ethylenediaminetetraacetate (EDTA) and/or heparin is centrifuged. The buffy coat is removed by aspiration, resuspended in plasma and recentrifuged in tubes made from pasteur pipettes. From such narrow columns buffy coat suspensions may be recovered virtually free of red blood cells (<6 per cent).

The recovery of nongranular leukocytes from EDTA-treated and heparin + EDTA-treated blood samples was 44.1 and 39.8 per cent respectively. The preparations obtained by both methods still contained high concentrations of thrombocytes. On an average, EDTA-treated and heparin + EDTA-treated preparations contained 230.3 and 58.4 thrombocytes per 100 leukocytes respectively. The recovery of thrombocytes in EDTA-treated preparations amounted to 42.7 per cent so that this technique may be used to concentrate thrombocytes for further study. The separation of buffy coat with heparin alone was not as clear and consistent because of the cell clumping which occurred.

     

Acute phase proteins in cattle: discrimination between acute and chronic inflammation.

Acute phase proteins such as serum amyloid A, haptoglobin, and alpha 1-acid glycoprotein have been identified as markers of inflammation in cattle because they are produced by the liver in response to pro-inflammatory cytokines. This study was designed to assess whether they could be used to discriminate between acute and chronic inflammation. Their concentrations were measured in serum samples from 81 cattle in which inflammation was classified by thorough clinical examination, supported by postmortem findings, as being acute in severity in 31 and chronic in 50. The classical haematological markers of inflammation were also determined in blood from the animals. Serum amyloid A had a maximum (100 per cent) clinical sensitivity in discriminating between the acute and chronic cases, and haptoglobin had the highest clinical specificity of 76 per cent; counts of neutrophils and band neutrophils had sensitivities of 71 per cent and 42 per cent and specificities of 30 per cent and 72 per cent, respectively. It was concluded that serum amyloid A and haptoglobin may be used to discriminate between acute and chronic inflammatory conditions.     

Anaemia and leucocytosis in one hundred and twelve dogs with pyometra

In a group of 112 dogs with pyometra, 31 per cent showed a normal red blood cell picture, 57 per cent non-regenerative normocytic normochromic anaemia and 12 per cent non-regenerative microcytic hypochromic anaemia. The non-regenerative anaemia was absent or moderate at lower white blood cell levels and markedly present at high to extreme white blood cell counts. The degree of non-regenerative anaemia was positively correlated with the degree of leucocytosis, neutrophilia, left shift and monocytosis. The more pronounced non-regenerative microcytic hypochromic anaemia, indicating a more severe chronic blood loss, was usually encountered at extremely high white blood cell levels. The significance of these observations is discussed.     

Comparison of the measurement of plasma testosterone and plasma oestrogens for the diagnosis of cryptorchidism in the horse

The results of performing 1720 blood tests for equine cryptorchidism are described. Using the paired sample human chorionic gonadotrophin (hCG) stimulation test and measuring testosterone, 6.7 per cent of tests did not give a clear result. If only the testosterone concentration in the pre-hCG blood sample was used, this percentage rose to 14 per cent. The paired sample hCG stimulation test was 94.6 per cent accurate. A comparison was made between the paired hCG stimulation test and the measurement of conjugated oestrogen in a single sample. The latter did not give as many doubtfuls but gave false negatives in donkeys of all ages and in horses less than three years old. Measurement of conjugated oestrogen was found to be 96 per cent accurate when donkeys and young horses were excluded. It is, therefore, recommended as a simpler and more reliable test than the paired sample hCG test, but only in the older horse.     

Acute phase proteins in serum and milk from dairy cows with clinical mastitis

The serum concentrations of haptoglobin, serum amyloid A and alpha1 acid glycoprotein were determined in serum collected from healthy dairy cows and cows with clinical mastitis, graded as mild (clots in milk) or moderate (clots in milk and visible signs of inflammation in the mammary gland/s) to assess their relative diagnostic value in detecting the disease. The concentrations of haptoglobin and serum amyloid A were also measured in milk collected from infected and uninfected quarters. The concentrations of haptoglobin and serum amyloid A were higher in the serum and milk from the cows with mild or moderate mastitis. The diagnostic value of haptoglobin in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 82 per cent and 94 per cent respectively with serum and 86 per cent and 100 per cent with milk. The diagnostic value of serum amyloid A in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 83 per cent and 90 per cent with serum and 93 per cent and 100 per cent with milk. The diagnostic value of serum alpha1 acid glycoprotein in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 62 per cent and 91 per cent.     

Haematological and biochemical measurements in healthy, adult, free-ranging golden jackals (Canis aureus syriacus) held in captivity

Blood from 31 healthy, free-ranging golden jackals held in captivity for seven days was collected while they were anaesthetised. Haematological and serum biochemical measurements were analysed and the 95 per cent confidence interval for each variable was compared with the reference value for domestic dogs. The measurements of their red blood cells were within the reference interval for dogs, but the jackals had higher white blood cell counts and eosinophil counts than dogs. The male jackals had a higher haematocrit, red blood cell count, mean corpuscular volume and mean corpuscular haemoglobin concentration, and a lower red blood cell distribution width than the female jackals. High activities of muscle enzymes were detected in many of the jackals, in several of which the activity of creatine kinase exceeded 5000 U/l; these were considered abnormal.     

Haptoglobin and ceruloplasmin as determinants of inflammation in dogs.

Assay procedures for determining serum haptoglobin concentration and ceruloplasmin oxidase activity in dogs were validated, and reference values were established. Serum haptoglobin concentration is reported as milligrams per deciliter of cyanmethemoglobin binding capacity, whereas serum ceruloplasmin oxidase activity was determined by use of p-phenylenediamine as substrate. Both assays were used to analyze serum samples from 288 dogs. In each dog's case record, clinical history and final diagnosis were evaluated to determine whether the dog had an inflammatory condition. Complete blood cell counts were performed in 265 dogs, using simultaneously collected blood samples. Plasma fibrinogen concentration was determined for 161 dogs. A positive correlation (P less than 0.01) for serum haptoglobin concentration and for ceruloplasmin oxidase activity, compared with WBC counts, segmented neutrophil and band neutrophil counts, and plasma fibrinogen concentration. Ceruloplasmin oxidase activity and haptoglobin concentration were up to 6 times more sensitive than fibrinogen concentration or leukocyte counts in detecting inflammation. Specificity of ceruloplasmin oxidase activity was comparable to fibrinogen concentration and leukocyte counts, whereas haptoglobin concentration was found to be slightly less specific. Specificity of haptoglobin concentration improved slightly (from 0.82 to 0.88) when dogs with a history of glucocorticoid administration were excluded from analysis. Predictive value of a negative test result (haptoglobin concentration less than 125 mg/dl; ceruloplasmin oxidase activity less than 20 IU/L) and predictive value of a positive test result for haptoglobin concentration and ceruloplasmin activity were comparable to or better than fibrinogen concentration or various oxidase leukocyte counts in detection of inflammation in a variety of disease conditions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epidemiological, clinical, haematological and biochemical characteristics of canine hypothyroidism

Hypothyroidism was diagnosed in 50 dogs and excluded in 86 dogs suspected of hypothyroidism, on the basis of the results of bovine thyrotropin response tests. Breed, pedigree, sex or neutering status did not significantly influence the likelihood of the dogs being hypothyroid. The hypothyroid dogs were significantly older than the non-hypothyroid dogs referred to the University of Glasgow during the same period. However, when dogs under two years of age were excluded from the statistical analyses there was no significant difference in age between the two groups. The most common clinical characteristics associated with hypothyroidism were metabolic signs (84 per cent of cases), particularly lethargy (76 per cent), obesity or weight gain (44 per cent), and exercise intolerance (24 per cent); and dermatological abnormalities (80 per cent), including alopecia (56 per cent), poor coat quality (30 per cent) and hyperpigmentation (20 per cent). When compared with the laboratory reference limits the most common biochemical and haematological abnormalities were increased concentrations of triglycerides (88 per cent), cholesterol (78 per cent), glucose (49 per cent), and fructosamine (43 per cent), and increased activities of creatine kinase (35 per cent), and decreased concentrations of inorganic phosphate (63 per cent), and a low red blood cell count (40 per cent). When compared with reference limits derived from the euthyroid dogs the most common abnormalities were increased concentrations of gamma-glutamyltransferase (21 per cent), cholesterol (18 per cent), and aspartate aminotransferase (15 per cent) and a decreased red blood cell count (29 per cent), and decreased neutrophils (18 per cent) and decreased activity of creatine kinase (15 per cent). Assessment of cholesterol, creatine kinase, aspartate aminotransferase, gamma-glutamyltransferase, and red blood cell and neutrophil counts may be particularly useful in distinguishing hypothyroid dogs from euthyroid animals with similar clinical signs.     

Assessment of a pyrogallol red technique for total protein measurement in the cerebrospinal fluid of dogs

The measurement of protein concentration in the cerebrospinal fluid is a basic analytical method in neurology. In this study, a pyrogallol red technique using a human albumin calibrator previously validated in human medicine was tested for canine samples, and the results were compared with those obtained using urine test strips. Pyrogallol red significantly (P<0.05) but moderately underestimated purified dog albumin and globulins. The imprecision of the technique was low: intra- and between-series coefficients of variation were 1.6 and 4.3 per cent at protein concentrations of about 0.3 g/litre. Over 49 samples, there was good agreement between the pyrogallol red and test strip results (r=0.63), especially for low and high protein concentrations, but misclassifications were observed with '+' test strip readings.     

Potassium concentration in equine red blood cells: Normal values and correlation with potassium levels in plasma

The concentration of potassium in plasma and in red blood cells was determined in 948 horses. The coefficient of correlation between the two parameters was low. In 436 of these horses, which were clinically healthy, the red blood cell potassium (RBCK⁺) levels did not fit within a normal distribution curve, but a bimodal distribution was observed with a section point at 90 mmol/litre. In 90 per cent of these normal horses, mean RBCK⁺ content was 97.5 mmol/litre. In the remaining 10 per cent, mean RBCK⁺ concentration was 93.8 mmol/litre. A subdivision into a ‘low potassium group’ and a ‘high potassium group’ was made. In 10 out of 15 horses in the ‘low potassium group’, bimonthly sampling over a period of one year showed that RBCK⁺ content remained low. In the remaining five horses an increase was observed.     

Changes in the blood biochemical and haematological profile of neonatal calves with age

Fourteen calves were used to investigate the changes from birth to 83 days of age in the concentrations of serum albumin, alkaline phosphatase, beta-hydroxybutyrate, plasma cortisol, serum creatine kinase, creatinine, iron, plasma fibrinogen, serum gamma-glutamyl transferase, plasma glucose, haptoglobin, serum non-esterified fatty acids, total protein, transferrin, triglycerides, urea and gamma globulin; the haematological variables measured were: basophils, eosinophils, haematocrit, haemoglobin, lymphocytes, mean cell haemoglobin, mean cell haemoglobin concentration, mean cell volume, monocytes, band neutrophils, neutrophils, platelets, red blood cells and white blood cells. The changes are presented as a series of graphs and the values are discussed in relation to the published reference ranges for adult cattle. Two populations of calves were identified which gave rise to a bimodal distribution for some of the variables. Differences in haematocrit, haemoglobin and red blood cell counts were apparent at birth, with raised values for these measurements being associated with an increased white blood cell and neutrophil count between three and 27 days of age.     

Distribution of T and B cells in thymus, blood and lymph nodes of the cat.

The proportions of T and B cells in the thymus, blood and lymph nodes were estimated in cats from three months to three years of age. T cells were identified by formation of E rosettes with guinea-pig red blood cells and B cells by the presence of surface immunoglobulin (Ig) as shown by the mixed antiglobulin reaction. Using a double test, it was shown that these methods identify separate, non-overlapping cell populations. The proportions of T cells shown in the thymus ranged from 10-73 per cent, in the blood 5-62 per cent and in the nodes, 5-44 per cent. Cells with surface Ig ranged from 0-10 per cent in the thymus, 26-68 per cent in the blood and 32-18 per cent in the nodes. Two cats with lymphadenopathy had unusually high B cell counts and one cat with depletion of the thymus was deficient in peripheral T cells. Papain treatment reduced or abolished E rosette formation by T cells.     

Differentiation of dogs with regenerative and non-regenerative anaemia on the basis of their red cell distribution width and mean corpuscular volume

The red blood cell distribution width (RDW), which provides a quantitative measure of the heterogeneity of the red cell population (anisocytosis) in the peripheral blood, the mean corpuscular volume (MCV) and a regression model combining both variables were used to assess their predictive accuracy in differentiating 51 dogs with regenerative anaemia from 92 dogs with non-regenerative anaemia, which had been diagnosed on the basis of the corrected reticulocyte count A classification tree analysis was constructed to generate an optimum set of diagnostic rules to differentiate between the two types of anaemia. Seventy-four dogs with a normal haemogram were used as controls. An increase of 1 per cent in the RDW and of 1 fl in the MCV increased the odds of an anaemic dog suffering from regenerative anaemia by factors of 1.3 and 1.14, respectively. By the classification tree, 78 per cent of anaemic dogs with a RDW of 16.25 per cent or less would be expected to have non-regenerative anaemia. With a RDW over 16.25 per cent, an MCV of 68.2 fl was the cut-off between dogs expected to have regenerative (71 per cent) or non-regenerative (75 per cent) anaemia. The RDW and MCV are measured by most automatic haematology analysers and may give the first indication of the bone marrow response of an anaemic dog. However, different electronic counters give different normal values of the RDW and MCV.     

A method for determining the concentration of haptoglobin in cattle blood following haemolysis caused at collection

This study aimed to develop an in-house assay for haptoglobin determination in bovine blood samples, assess the effect of haemolysis on the reported haptoglobin concentration and develop a method to correct for haemolysis interference. The assay developed is highly repeatable (92.3% across plates and 94.8% between assays). A correction equation (Hp(corrected)=Hp(raw)-Hp(endogenous activity)-Hp(due to Hb); where Hp(due to Hb)=0.118×Hb(free)+0.015) was developed based around the linear relationship of haptoglobin and haemoglobin (by-product of haemolysis) and endogenous interference, tested and validated for use with haemolysed samples. The method described in this paper allows samples inadvertently haemolysed at collection to be analysed, with the reported haptoglobin concentration being an accurate reflection of the physiological levels in the animal's blood at the time of collection.     

Effect of clinoptilolite on serum biochemical and haematological characters of broiler chickens during aflatoxicosis

Clinoptilolite (CLI, a natural zeolite) incorporated into the diet at 1.5 and 2.5 per cent were evaluated for their ability to reduce the deleterious effects of 2.5 mg total aflatoxin (AF) kg(-1)diet on broiler chickens from 1 to 21 days of age. In total 360 broiler chicks were divided into six equal treatment groups (control, AF, CLI (1.5 per cent), AF plus CLI (1.5 per cent), CLI (2.5 per cent) and AF plus CLI (2.5 per cent)). When compared with the controls, AF treatment significantly decreased serum total protein, albumin, inorganic phosphorus, uric acid, total cholesterol and the values of haematocrit, red blood cell counts, mean corpuscular volume, haemoglobin, thrombocyte counts, percentage of monocyte counts; increased values of white blood cell and heterophil counts. The addition of CLI (1.5 per cent) and CLI (2.5 per cent) to the AF -containing diet reduced the adverse effects of AF and should be helpful in a solution to the aflatoxicosis problem in poultry.     

Isolation of equine peripheral blood mononuclear cells using Percoll.

The concentration of Percoll required for isolating equine peripheral blood mononuclear cells has been reinvestigated. A poor cell yield was obtained at the 60 per cent concentration already reported. It is recommended that workers specifically interested in high yields of mononuclear cells, for investigation of lymphocyte and monocyte functions, use a concentration of 65 per cent Percoll. However, workers wishing to isolate pure populations of equine neutrophils might consider a concentration of 70 per cent in the upper layer of Percoll used to retain the mononuclear cells.     

Haematological responses of repeated large volume blood collection in the horse

The haematological response of regular, repeated blood harvesting was investigated in 40 Thoroughbred and non-Thoroughbred horses that donate 8 litres of blood every 3 weeks for the purposes of commercial blood production. When this volume of blood was removed on five occasions over 12 weeks, no adverse effect on packed cell volume (PCV), haemoglobin (HB), and red blood cell count (RCC) was observed. Although PCV, RCC and Hb values decreased during the first week after blood collection, followed by a gradual increase in values until the next harvest time, all values remained within published reference ranges. Derived red cell indices [mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), and mean corpuscular haemoglobin concentration (MCHC)] also remained within reference range. We conclude that the removal of approximately 8 litres of blood (approximately 16 ml kg(-1)or 20 per cent of blood volume for a 500 kg horse) from blood donor horses every 3 weeks allows time for adequate recovery of haematological variables and does not result in adverse haematological changes.     

Distribution and ultrastructure of mast cells in the equine respiratory tract

Mast cells in the equine respiratory mucosa were studied at both light--and transmission electron--microscope levels. Mast cells were identified at all levels of the tract, with the greatest cell density in the nasopharynx. The majority (57 to 94 per cent) of this cell population were located within the connective tissue of the lamina propria. Up to 20 per cent of these cells were associated with the mucosal glandular tissue, whilst small numbers were present within the surface epithelium and in association with nodular lymphoid tissue. In the peripheral lung tissue 20 per cent of the mast cell population was associated with the small airways, 25 per cent with the pleura, and 32 per cent with blood vessels. The fine and ultrastructural features of these mast cells resemble those described in other species, and their granules consist of two types which resemble human mast cells.