Immunologic Profiles of Peripheral Blood Leukocytes and Serum Immunoglobulin G Concentrations in Perinatal Mares and Neonatal Foals (Heavy Draft Horse)

The purpose of this study was to examine sequential changes in the immunologic parameters of perinatal mares and neonatal foals of the heavy draft horse. Blood samples were collected from clinically healthy pregnant mares and their newborn foals every week from 1 month before the expected foaling date, and 1 hour, 1 day (24-48 hours), and 1, 2, 3, and 4 weeks after foaling. Peripheral blood samples were used to examine total leukocyte counts (n = 20), differential leukocyte counts (n = 20), lymphocyte subpopulations (n = 13), lymphocyte responses to mitogens (n = 10), neutrophil phagocytic function (n = 12), and serum immunoglobulin G (IgG) concentrations (n = 10). In perinatal mares, remarkable changes observed included increased neutrophils, decreased lymphocytes, decreased CD4⁺ T lymphocytes, and decreased lymphocyte responses to mitogens at delivery. These changes were speculated to be the result of physical stress associated with delivery. In neonatal foals, increase in the phagocytic function of neutrophils, and increase in serum IgG concentration after suckling colostrum and increase of lymphocytes accompanied by physiologic growth were observed. Compared to dams, foals showed lower phagocytic function of neutrophils before suckling and fewer lymphocytes and lower lymphocyte responses to mitogens within 1 day after birth. This study revealed immunologic dynamics in perinatal mares and neonatal foals. Immunologic functions are suppressed in foaling mares and are immature in neonatal foals, especially before colostral intake. We expect these data will be useful for further studies in the field of clinical immunology, and preventive medicine.     

Changes in peripheral blood leucocyte counts and subpopulations after experimental infection with BVDV and/or Mannheimia haemolytica

Leucocyte counts and subpopulations were studied in peripheral blood from calves experimentally infected in the respiratory tract with either bovine virus diarrhoea virus (BVDV) or Mannheimia haemolytica (Mh), or with a combination of both agents (BVDV/Mh). A non-inoculated control group was included. Peripheral blood samples were obtained for total leucocyte counts, and for neutrophil, lymphocyte and monocyte counts. The numbers of blood lymphocytes expressing the surface antigens CD4, CD8, WC1, B and IL-2R were analysed using flow cytometry. The results showed that BVDV inoculation induced a significant decrease in total leucocyte counts and in neutrophil and lymphocyte numbers, while Mh inoculation induced significant increases in total leucocyte counts and neutrophils, while the lymphocyte count decreased. In the BVDV/Mh group, the total leucocyte count and the lymphocyte numbers decreased significantly. In this group, the lymphocyte numbers remained on a very low level throughout the rest of the study. The numbers of CD4+, CD8+ and WC1+ lymphocytes decreased significantly compared with before inoculations mainly in the BVDV and BVDV/Mh groups. The drops were most pronounced in the BVDV/Mh group. The numbers of B+ lymphocytes and IL-2R+ cells did not change significantly.     

Functional and ultrastructural evaluation of neutrophils from foals and lactating and nonlactating mares

Neutrophils from 4 pony foals, 3 lactating pony mares, and 3 nonlactating mares were evaluated ultrastructurally and by in vitro function tests. Neutrophils from foals had significantly (P = 0.05) less random migration than neutrophils from mares; values in tests for iodination and Staphylococcus aureus ingestion were also lower with foal neutrophils. Neutrophils from lactating mares had lower responses to iodination, antibody-dependent cell-mediated cytotoxicity, and random migration tests than did neutrophils from nonlactating mares. Ultrastructurally, granule concentration did not differ significantly among groups. A slight decrease in primary granules and a corresponding increase in granules with a flocculent matrix indicates partial spontaneous neutrophil degranulation in foals and lactating mares.     

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.     

Leucocyte counts in the healthy English Thoroughbred in training

Total and differential leucocyte counts have been determined by electronic counting techniques in 474, two-to four-year-old healthy Thoroughbreds in training. The ranges of values observed (particularly those for neutrophils and lymphocytes) were narrower than previously described. Absolute and percentage lymphocyte values were significantly decreased with advancing age causing a relative increase in percentage neutrophils. The fall in absolute lymphocyte numbers was the main cause for a significant decrease in total leucocytes with age. Frequency plots for each variable showed that, with the exception of the eosinophil count, data from other leucocyte cell types following either a normal or lognormal distribution.     

Flow cytofluorometric characterization of bovine blood and milk leukocytes

Flow cytometry and sorting proved to be a rapid method that facilitated the identification of different leukocyte populations in bovine blood and milk. After briefly incubating whole blood and milk samples in a hypotonic phosphate buffer, containing supravital acridine orange, 5 classes of leukocytes were found in the blood (lymphocytes, neutrophils, eosinophils, basophils, and monocytes) and 4 in the milk (lymphocytes, neutrophils, monocytes, and macrophages) by flow cytometry. Cells were morphologically identified by fluorescent microscopy after flow cytometric sorting and by light microscopy after Papanicolaous staining. Udder parenchymal and ductal tissue cells (secretory and epithelial cells) were not found in the milk samples evaluated. Large differences in the total and differential cell counts were found in the different milk secretions.     

Cytology of haematological cells of otariid seals indigenous to Australasian waters

OBJECTIVE: To characterise the morphology of cells from the blood of otariid seals. METHODS: Examination of blood by light and scanning electron microscopy. RESULTS: Erythrocytes were typically discocytes. Leukocytes that were recognised included neutrophils, lymphocytes, monocytes, eosinophils and basophils. The morphology varied little between species for neutrophils, lymphocytes and monocytes. Some variation between species was observed for eosinophils and basophils. Platelets were generally aggregated. CONCLUSION: This study provides a guide for the identification of cells from the blood of otariid seals.     

Use of flow cytometry for determination of differential leukocyte counts in bovine blood

A flow cytometric method was developed to perform differential leukocyte counts on bovine blood. Blood specimens from 50 healthy Holstein cows were analyzed by use of a flow cytometer. The method entailed diluting blood with phosphate-buffered, hypotonic saline solution containing acridine orange, and performing a step-wise, 3-parameter analysis on the bases of cell size, cellular granularity, and granulocyte fluorescence. Initially, proportions of monocytes, granulocytes, and lymphocytes were determined by creating appropriate windows on dot plots of cell size (determined by forward light scatter) vs cellular granularity (determined by the logarithm of side light scatter). Eosinophils were resolved by analysis of granulocytes as dot plots of logarithms of green vs red fluorescence ascribed to acridine orange. Proportions of eosinophils and neutrophils were computed from data so generated. Microclumps of platelets spuriously affected counts of some granulocytes, particularly eosinophils. Differential leukocyte counts determined by flow cytometry generally compared favorably with those obtained by use of the conventional microscopic method, using Wright-stained blood films. Mean neutrophil and eosinophil counts determined by the 2 methods did not differ significantly, but lymphocyte counts determined by flow cytometry were significantly higher than those determined by microscopy (P less than 0.01). Correlation coefficients for counts of neutrophils, eosinophils, and lymphocytes determined by the 2 methods ranged from 0.519 to 0.833. Correlation between monocyte counts was low (r = 0.147), although mean monocyte counts determined by the 2 methods did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of altrenogest‐treatment of mares in late gestation on adrenocortical function,blood count and plasma electrolytes in their foals

Reasons for performing study: Mares with compromised pregnancies are often treated with altrenogest to prevent abortion. However, there is only limited information about effects on the foal when altrenogest treatment is continued during final maturation of the fetus. Objectives: To determine effects of altrenogest treatment during late gestation in mares on maturity, haematology changes, adrenocortical function and serum electrolytes in their newborn foals. Methods: Six mares were treated with altrenogest (0.088 mg/kg bwt) once daily from Day 280 of pregnancy until foaling and 7 mares served as controls. Results: Foals born to altrenogest‐treated mares had a significantly lower neutrophil/lymphocyte ratio on the first day after birth than control foals (P<0.05). Basal plasma cortisol concentrations immediately after birth were higher in foals of altrenogest‐treated mares than in control foals (P<0.05). Cortisol release in response to exogenous adrenocorticotropic hormone (ACTH) ‐ except for higher values 15 min after ACTH injection in foals of altrenogest‐treated mares on Day 1 ‐ revealed no differences in adrenocortical function between the groups of foals. Plasma potassium concentration in foals from altrenogest‐treated mares compared to control foals was significantly lower immediately after birth (P<0.05) and plasma ionised calcium concentration was significantly lower 3 h after birth (P = 0.01). Conclusions and potential relevance: Altrenogest treatment of pregnant mares prolonged labour had no major effects on adrenocortical function in foals. A reduced neutrophil/ lymphocyte ratio in these foals may suggest either immunomodulatory effects of altrenogest or dysmaturity of the foals.     

The Effects of Selenium Source on Measures of Selenium Status of Mares and Selenium Status and Immune Function of Their Foals

The effect of organic and inorganic sources of selenium (Se) on measures of Se status of mares and their foals, Se concentrations of colostrum and milk, and indices of immune function of foals was studied. Twenty late-gestation Standardbred mares were randomly assigned to one of two groups. All mares received an identical balanced ration, except for the source of supplementary Se: one group received organic Se (Se yeast) and the other group received inorganic Se (sodium selenite), fed to deliver 0.3 mg/kg supplementary Se based on total ration dry matter. Mares received the experimental diet from 2 months before estimated due date until 1 month after foaling. Source of Se did not affect Se concentrations in maternal plasma, red blood cells, colostrum, or milk. At 1 month of age, foals from mares fed organic Se had higher red blood cell Se concentration than foals from mares fed inorganic Se (P < .05). Measures of immunity included serum immunoglobulin G concentration, lymphocyte proliferation in response to concanavalin A, and relative cytokine gene expression of stimulated lymphocytes (interferon gamma [IFNγ], interleukin [IL]-2, IL-5, IL-10, tumor necrosis factor alpha [TNFα]) and neutrophils (IL-1, IL-6, IL-8, IL-12, TNFα). Relative gene expression of IL-2, TNFα, and IFNγ by foal lymphocytes was associated with the source of Se supplementation provided to the mares. We conclude that the source of dietary Se provided to mares may influence the immune function of foals at 1 month of age through changes in relative gene expression of certain lymphocyte cytokines.     

Determination of haematological reference intervals in healthy adult greyhounds

Objective : The purpose of this study was to establish breed‐specific reference intervals for haematological measurands in non‐racing greyhounds. Suitability of the data for partitioning according to sex was also examined. Methods : Haematological data were collected from 304 healthy non‐racing greyhounds and analysed using non‐parametric methods. Results were compared with non‐breed‐specific canine reference intervals and also with greyhound reference intervals obtained by other investigators. Results : Compared with non‐breed‐specific reference intervals, the results showed comparable mean and upper limit and higher lower limit for erythrocyte count; higher values for haemoglobin, haematocrit and mean corpuscular volume; and lower values for total leucocyte count and absolute concentration of neutrophils, lymphocytes, monocytes, eosinophils and platelets. Partitioning according to sex was recommended by the statistical analysis for all analytes except haematocrit and total leucocyte count. Clinical Significance : In this study the reference intervals were derived from a large sample size. The results are in general agreement with previous reports, although higher values for low reference limits have been noted for the erythroid parameters, and lower values for upper reference limits have been observed for the total and different leucocyte counts. Breed‐specific reference intervals provide a useful clinical tool for haematological evaluations.     

Morphology of leukocytes from cats affected with alpha-mannosidosis and mucopolysaccharidosis VI (MPS VI)

The morphology and ultrastructure of circulating white blood cells from six Persian and from five Russian Blue/Siamese cats deficient in lysosomal activity of alpha-mannosidase and arylsulfatase B, respectively, were studied and compared to cells from corresponding normal and carrier cats. In cats with mannosidosis, light microscopic examination revealed vacuoles in lymphocytes and monocytes, whereas electron microscopic studies demonstrated additional vacuoles in neutrophils, eosinophils, and basophils. In cats with mucopolysaccharidosis VI (MPS VI), vacuoles containing metachromatic granules were observed in lymphocytes, neutrophils, eosinophils, and monocytes. Ultrastructural studies of these cells identified the accumulation of fibrillar material, which often was associated with lamellated membrane structures.     

Effect of subcutaneous injection of ginseng on cows with subclinical Staphylococcus aureus mastitis

Cows with subclinical mastitis caused by Staphylococcus aureus were subjected to subcutaneous injections with either an extract from the root of Panax ginseng CA Meyer at a dose of 8 mg/kg body weight per day for 6 days, or with saline as a control. The injection areas were checked for adverse reactions. The daily milk production was measured before and after treatment. Blood was collected for total and differential leucocyte counts, identification of lymphocyte subpopulations using flow cytometry, lymphocyte proliferation test, and neutrophil phagocytosis and oxidative burst assay. Quarter milk samples were collected for bacteriological analysis and somatic cell counts (SCC). After the end of treatment, the numbers of S. aureus-infected quarters and milk SCC tended to decrease in ginseng-treated cows. Phagocytosis and oxidative burst activity of blood neutrophils were significantly increased 1 week after ginseng treatment, but the proliferative response of blood lymphocytes did not change significantly. The number of monocytes in ginseng-injected cows was significantly higher 1 week post-treatment than pre-treatment, and the number of lymphocytes was significantly higher than pre-infusion at 2 and 3 weeks after ginseng treatment. Similar changes were not observed in the control group. The present findings indicate that ginseng treatment can activate the innate immunity of cows and may contribute to the cow's recovery from mastitis. It is therefore suggested that ginseng has a potential as a stimulator of the immune system of dairy cows.     

Blood constituents of farmed red deer (Cervus elaphus): I. Haematological values

Approximately 100 farmed male and female red deer aged three months and over were blood-sampled and haematological parameters were measured. The deer were sampled by jugular venepuncture without tranquillization. Mean values were haemoglobin (16.0g/dl), packed cell volume (44.6%), mean corpuscular haemoglobin concentration (35.8g/dl), total plasma protein (66.0g/l). fibrinogen (4.8g/l) and white blood-cell numbers (5.80 x 10⁹/l). Differential white blood-cell counts were neutrophils (53.9%), eosinophils (4.8%), basophils (3.0%), lymphocytes (37.2%) and monocytes (1.1%). There were no significant differences in any parameter measured between sexes or between age groups three to eight months, nine to eighteen months, or older.     

Age-related changes in selected haematology parameters in rabbits

Even though there is an abundance of information on the reference values of haematological parameters in adult rabbits, a little is known about the changes in haematology in newborn rabbits or during their postnatal development. Therefore, the aim of our study was to investigate changes in red blood cells (RBC), white blood cells (WBC) and differential leukocyte counts in SPF New Zealand White rabbits from the age of one day to 20 weeks. Significant age-related changes during the first four weeks of life were detected. These included an increase of RBC and WBC, reversal of the neutrophil/lymphocyte ratio and increase of total counts of eosinophils and basophils. From the age of six weeks of life, all of the studied haematological parameters were comparable to those of adult rabbits.     

Effects of dietary Centella asiatica (L.) Urban on growth performance,nutrient digestibility,blood composition in piglets vaccinated with Mycoplasma hyopneumoniae

To investigate the effects of Centella asiatica (L.) on growth performance, nutrient digestibility and blood composition in piglets, 32 nursery pigs were fed 0.0, 0.5, 1.0 and 2.0% dietary C. asiatica (L.) from 15 to 90 kg BW. At 30 kg BW, nutrient digestibility was measured and at 35 kg BW piglets were vaccinated with Mycoplasma hyopneumoniae. Hematological parameters were checked at 40 and 80 kg BW. Compared with the control, growth performance was not affected. The ether extract, ash and calcium digestibility were lower at 0.5%, and dry matter, crude protein, crude fat, phosphorus and energy digestibility were lower at 1.0% (P < 0.05). On hematological values, at 40 kg hematocrit, total white blood cells, neutrophils, eosinophils, basophils, monocytes and lymphocytes were higher at the 2.0% level (P < 0.05). Most of these values except basophils and monocytes continued until at 80 kg, at which total white blood cells, neutrophils, eosinophils and lymphocytes were higher even at 1.0% (P < 0.05); neutrophil‐to‐lymphocyte ratio tended to be higher at 2.0% (P < 0.03). Cholesterol, triglycerides and antibody levels against M. hyopneumoniae did not differ except that at 40 kg the cholesterol of 0.5% was lower (P < 0.05) and M. hyopneumoniae‐specific antibodies tended to be higher with increasing levels of C. asiatica (L.) (P < 0.07). The result that C. asiatica (L.) could not improve growth performance but increased values of serum hematocrit and white blood cells, and mycoplasma immunity to M. hyopneumoniae might suggest that C. asiatica (L.) has no function to elevate body weight but has the potential to enhance innate immunity.     

Haematological and immunological variables in a domesticated and wild subspecies of ostrich (Struthio camelus)

1. Domesticated ostriches have been selected rigorously for productive traits with little concern for immunological responses, in contrast to wild ostriches. 2. We hypothesised that the immunological responses of wild and domesticated ostriches would differ. Total leucocyte counts, differential counts, heterophil: lymphocyte ratios, phagocytic activity, lysosome levels and anti-sheep red blood cell (SRBC) antibody titres (total, IgG, IgM) were compared between domesticated (n=3) and wild (n=3) ostrich subspecies. 3. Total leucocytes, lymphocytes and heterophils were similar in the 2 subspecies, but basophils and eosinophils were lower in the wild than in the domesticated ostriches. Lysosome concentrations and phagocytic activities were higher in the wild ostriches. 4. Total and IgM antibody titres to SRBC reached peak values quicker in the domesticated than in wild ostriches. IgG development patterns were similar. 5. The results suggest that a stronger non-specific immune response was shown by the wild ostriches (higher phagocytosis and lysozymes) whereas a stronger specific immune response was shown by the domesticated ostriches (peak values of anti-SRBC antibody titres were reached more quickly).     

Hematologic, biochemical, blood-gas, and acid-base values in greyhounds before and after exercise

After racing 722 m, 16 Greyhounds were evaluated to determine changes in hematologic, biochemical, blood-gas, and acid-base values following exercise. Values were determined before racing (T0), immediately after racing (T1), and 3 hours after racing (T2). Significant changes detected immediately after racing included increased heart rate, respiratory rate, and rectal temperature. Significant changes in hematologic values included increases in PCV, total plasma protein, hemoglobin, RBC, WBC, neutrophils, and lymphocytes. Change was not detected in values for monocytes, eosinophils, and neutrophil/lymphocyte ratio. Other increases included those for plasma concentrations of sodium, chloride, calcium, lactic acid, aspartate transaminase, alanine transaminase, alkaline phosphatase, creatine kinase, lactate dehydrogenase, and glucose. Concentrations of potassium and urea did not change. Measurement of blood-gas and acid-base status revealed significant increases in PaO2 and base deficit, whereas PaCO2, pH, and bicarbonate decreased. Three hours after exercise, all vital signs and blood-gas and acid-base values, except for PaCO2, which was still slightly low, had returned to baseline (T0) values. Most biochemical values had also returned to baseline, although sodium, chloride, aspartate transaminase, and creatine kinase were still high, and urea was low. Many hematologic values were still different from baseline values, with high values for WBC, neutrophils and neutrophil/lymphocyte ratio, and low values for PCV, total plasma protein, hemoglobin, RBC, and lymphocytes.     

Cytological and bacteriological findings in guttural pouch lavages of clinically normal horses

Percutaneous washes of the guttural pouches were obtained from two groups of 15 clinically normal horses, one lightly exercised and the other heavily exercised. Microbiological and cytological studies showed a wide variation in the differential cell counts. The cytological pattern of the normal lavages (< 5 per cent neutrophils) was characterised by a large proportion of ciliated columnar epithelial cells, a few non-ciliated cuboidal epithelial cells, and less than 1 per cent monocytes, lymphocytes, and eosinophils. Abnormal lavages (with more than 5 per cent neutrophils) had higher levels of bacterial growth than normal lavages. There were significant differences between the bacterial growth and total cell count, and also between the neutrophil contents of the lavages from the two groups of horses.     

Functional and ultrastructural changes in neutrophils from mares and foals experimentally inoculated with a respiratory tract strain of equine herpesvirus-1

Neutrophils isolated from venous blood of adult and foal ponies inoculated with equine herpesvirus-1 were evaluated by in vitro function tests and by electron microscopy. Foals had fever and severe neutropenia 24 hours after inoculation; increased neutrophil random migration under agarose and decreased antibody-dependent cell-mediated cytotoxicity were significant at 24 hours, but values had returned to preinoculation levels by 72 hours. Mares had fever and leukopenia of less severity, increases in neutrophil migration, and longer persistence of primary granule release than were seen in foals. Reduced migration and degranulation, and a decrease in antibody-dependent cell-mediated cytotoxicity seen with neutrophils from foals, as compared with mares, may relate to the high susceptibility of foals to equine herpesvirus-1 infection.