Perspectives and advances in in-clinic laboratory diagnostic capabilities: hematology and clinical chemistry.

The typical technologies used in veterinary hematology and biochemical analyzers are reviewed, along with associated advantages and disadvantages. Guidelines for implementing a successful in-clinic laboratory are provided, including criteria for system evaluation and expectations for comparative performance evaluations. The more common problems and limitations associated with in-clinic laboratory diagnostics and how to best prevent them are also discussed.     

Hematology, blood typing, and immunology of the neonatal foal

Hematologic parameters change during the first 10 days of life. Erythrocytes increase in number but decrease in size and hemoglobin concentration. The PCV, hemoglobin, and platelet count also decrease. Total blood and plasma volume and, to lesser extent, erythrocyte volume decrease. Normal neonatal foals may have immature neutrophils (up to 5 per cent bands), and their early rapid rise in neutrophil numbers may be accompanied by a lymphopenia. Monocytes, eosinophils, and basophils are all absent or low initially. Infectious processes can cause rapid and variable changes in the leukogram. However, elevation of fibrinogen levels may lag behind the development of an inflammatory process, and this parameter should not be relied on for early evidence of infection. After 12 hours of life, there is generally a decrease in serum concentrations of Na, Cl, iron, creatinine, BUN, plasma protein, and possibly calcium. LDH, SAP, P, bilirubin, and glucose concentrations are all higher in foals than in mature horses. Creatinine may actually be elevated during the first 12 hours of life and then decreases. If azotemia, hypochloremia, hyponatremia, and hyperkalemia are found, ruptured bladder with uroperitoneum should be suspected. The creatinine concentration is preferable to BUN determination for diagnosis of this condition. Blood typing is useful for diagnosis of NI, determination of blood compatability between donor and transfusion recipient, and for verification of parentage for breed registries. Several techniques are available. Several tests are available for evaluation of the foal's immunoglobulin levels and confirmation of passive antibody transfer. Because foals suffering from FPT are more predisposed to infections, their immunoglobulin status should be determined as early as possible so that additional colostrum or plasma can be administered as needed. Neonatal isoerythrolysis is uncommon but is an important immunologic syndrome that often results in a fatal hemolytic crisis. If one suspects the condition may be likely, the optimal time for testing the mare is during the last 2 weeks of gestation. If the foal's dam is shown to have alloantibodies against a panel of known erythrocyte alloantigens, prevention is possible by feeding colostrum from another mare. If a foal develops NI, further colostrum ingestion from the dam must be prevented. Good nursing care, minimizing stress, and adequate frequent feedings are essential; prophylactic antibiotics should be used, and transfusion may be necessary.     

Hematology of the sandbar shark, Carcharhinus plumbeus: standardization of complete blood count techniques for elasmobranchs

BACKGROUND: Standardized hematologic methods and reference intervals have not been established for cartilaginous fishes (sharks, skates, and rays) despite the large number of animals displayed in zoos and aquariums worldwide. OBJECTIVE: The focus of this study was to validate CBC methods for sandbar shark (Carcharhinus plumbeus) blood, based on criteria established in human medicine, for the following tests: RBC count, total WBC count, PCV, hemoglobin (Hgb) concentration, and WBC differential percentages. METHODS: Replicate CBCs were performed using blood samples from 5 captive sandbar sharks. Three protocols for RBC and total WBC counts were compared, as were different centrifugation times for PCV determination, and 2 methods for Hgb concentration. Means, minimum and maximum values, and CVs were compared to CAP and CLIA performance guidelines for human tests. RESULTS: Total WBC counts in a diluent modified for elasmobranch blood, Hgb concentration by the cyanmethemoglobin method after removal of nuclei, and WBC differential percentages showed acceptable performance. PCV results were acceptable when tubes were centrifuged for at least 5 minutes. Total RBC counts by all 3 methods exceeded the acceptable error for manual counts of human cells. CONCLUSIONS: Standardized CBC tests can be used as health assessment tools for elasmobranchs. Total RBC counts should be viewed as estimates.     

Hematology and biochemistry reference values for the ranch fox.

Reference hematology and biochemistry values are presented from a mixed population of 30 silver and red foxes of both sexes, reared and living under fox-farming conditions in southern Ontario. Based on history and physical examination, the animals in this study were clinically healthy at the time of blood collection and maintained under similar husbandry practices. The observations were examined for outliers and Gaussian distribution before and after one of three transformations. Parametric analysis was used to determine lower and upper reference limits. Where observations were not Gaussian, minimum and maximum values are given. These reference values are presented as a usable first approximation of population reference values to assist clinicians and researchers in their interpretation of observations obtained from foxes of similar populations.     

Pathology of the mononuclear cell leukemia of Fischer rats. II. Hematology

Complete hemograms were evaluated for 57 rats with mononuclear cell leukemia and compared to hemograms obtained from 52 age- and sex-matched nonleukemic rats. All leukemic rats had marked hemolytic anemia and associated spherocytosis, reticulocytosis, anisocytosis, and polychromasia. The anemia varied with the stage of illness and was more severe in rts with advanced leukemia. Death appeared to be related to anemia. There was a marked neutrophilia with left shift, mild lymphopenia, and moderate to severe thrombocytopenia. Atypical mononuclear cells were detected in circulation in all but three rats. Total white blood cell counts ranged from 5.0-370 x 10(3) cells/ml. There was an increase in erythrocyte osmotic fragility with separation into two distinct populations of erythrocytes. Eight of nine rats were Coombs' positive indicating an immune-mediated pathogenesis for the anemia. Hemostasis tests revealed a markedly prolonged prothrombin time, hypofibrinogenemia, slightly increased to normal partial thromboplastin time, and undetected fibrin degradation products. These findings suggest significant liver disease associated with the leukemia.     

Hematology of the neonatal calf. III. Frequency of congenital iron deficiency anemia.

The hematocrits of 258 Holstein-Friesian and Jersey calves were determined during the first 3 days following birth. Hematocrit values of 25% or less were observed in 15.8% of the calves and hematocrits of 20% or less were observed in 6.9% of calves from apparently normal dams. The serum iron concentration of neonatal calves with hematocrits of 25% or less was significantly lower than the value recorded in calves with hematocrits above 25% and there was a direct correlation between PCV and serum iron concentration. The data presented suggested that prenatal development of iron deficiency is an important cause of anemia in neonatal dairy calves. Factors which may be important in pathogenesis are discussed.     

Altered monocyte and macrophage numbers in blood and organs of chickens injected i.v. with lipopolysaccharide

Lipopolysaccharide (LPS) is a Gram-negative bacteria cell wall component that activates monocytes and macrophages to produce nitric oxide (NO) from inducible nitric oxide synthase. Nitric oxide production in the plasma of chickens peaks 5–6-h post-i.v. LPS injection reflecting iNOS activation. To determine monocyte responsiveness after an i.v. LPS injection, a time course study was conducted examining the concentrations among peripheral blood leukocytes post-i.v. LPS injection in male and female chickens, the proportions among peripheral mononuclear leukocyte (PBMC; containing lymphocytes, thrombocytes, and monocytes) populations isolated from the blood samples collected at various times post-i.v. LPS treatment, and the ability of monocytes to produce NO with and without further LPS stimulation in vitro using the PBMC NO production assay. Additionally, monocyte extravasation activity was determined by analyzing macrophage proportions after the i.v. LPS injection in spleen, lung, and liver tissues. Blood was collected from male and female chickens at 0 h (pre-LPS injection control) and at 1, 3, 6, 24, and 48 h post-LPS injection, and additionally, at 72 h from female chickens. Tissues were collected 0, 1, 6, and 48 h post-i.v. LPS injection from male chickens. Monocyte concentrations dropped substantially by 1 h in both males and females. In males, monocyte concentrations returned to control concentrations by 6 h and increased at 24- and 48-h post-LPS injection, whereas in females, monocyte concentrations recovered more slowly, returning to near control concentrations by 24–48-h and increasing above control levels by 72 h. Lipopolysaccharide stimulated NO production by PBMC cultures established from blood samples obtained at various times post-LPS injection in vivo followed the same pattern as monocyte concentrations in the blood. Hence, NO concentrations within PBMC cultures were dependent upon the number of monocytes that were in the PBMC cultures isolated at different times post-i.v. LPS injection. Furthermore, macrophage proportions in spleen tissues responded similarly to monocyte concentrations in the blood, decreased in lung tissue, and varied widely in liver tissue throughout 48 h after an LPS injection. Monocytes and other leukocytes may attach to the endothelium post-i.v. LPS injection preventing the monocytes from entering the needle during blood collection resulting in what seems to be leukopenia in blood and in PBMC cultures attenuating NO production in PBMC cultures. Furthermore, monocyte differentiation and recruitment from the bone marrow is a likely contributor to the reconstitution and rise of monocyte concentrations in blood samples post-i.v. LPS injection.     

Hematology, blood chemistry, and bacteriology of the free-ranging Mexican beaded lizard (Heloderma horridum).

Blood samples from 16 adult, wild Mexican beaded lizards were obtained for complete blood count and serum biochemical analysis. In addition, both oral and cloacal swab samples were collected and submitted for bacteriologic culture. This is the first report of hematologic and bacterial data from free-living beaded lizards. This information will serve as baseline reference values for future health assessment studies in conservation, captive breeding, and research programs.     

Hematology and serum biochemistry of Holstein dairy calves: age related changes and comparison with blood composition in adults

Specific reference intervals are needed for each animal species for appropriate interpretation of hematological and serum biochemical results. The aim of the present study was to investigate the blood composition of growing calves in order to evaluate the need for defining reference values for different age groups. Thirty two Holstein calves (18 male and 14 female) were blood sampled. A blood sample was taken within 24-48 h following birth and at 14, 28, 42, 56, 70 and 84 days of age. CBC determination and the measurements of some blood serum metabolites, enzymes, electrolytes and minerals were performed. There were significant age related changes for most hematological and biochemical parameters (p < 0.05) except for the numbers of band neutrophils and monocytes and the amounts of sodium, potassium, chloride and BUN. The results of the present study showed that for some hematological and biochemical parameters such as hemoglobin, MCV, MCH, MCHC, inorganic phosphorus, serum total protein, globulin, AST and ALP at the first three months of life and also, neutrophil numbers and glucose levels at the 24-48 h of life, the age specific reference values must be considered for precise interpretation of laboratory results.     

Clinical evaluation of blood lactate levels in equine colic.

Blood lactate levels were evaluated in 36 horses (43 cases) presented with colic. A correlation between increasing blood lactate levels and decreasing percentage survival has been shown. An appreciable anion gap was found in 7 of 10 cases analyzed in detail but in each case the entire gap could not be accounted for by lactate alone. Proposals are offered to account for the unmeasured anions. Blood lactate determination is suggested as a prognostic rather than a diagnostic aid for the equine practitioner and should be used to augment other clinical findings in the horse exhibiting colic.     

Hematology of natural bovine trypanosomosis in the Brazilian Pantanal and Bolivian wetlands.

We report hematological changes observed in natural cases of bovine trypanosomosis due to Trypanosoma vivax in beef and dairy cattle from Bolivian wetlands and Pantanal, Brazil. The main hematologic changes produced by T. vivax infections were anemia and severe leucopenia. The cattle presented macrocytic hypochromic anemia. The leukocyte changes were characterized by relative lymphocytosis and monocytosis and decrease in the neutrophil counts. The clinical signs were lachrymation, progressive weakness, marked weight loss, inappetence, diarrhea and abortions during the third trimester of pregnancy.     

Big decisions based on small numbers: lessons from BSE.

The epidemic of bovine spongiform encephalopathy (BSE) has been the most expensive disaster ever to have befallen farming in the UK. It is believed to have led to a new form of spongiform encephalopathy in humans and as yet there is no way of knowing how many people will die of this disease. In order to curtail the BSE epidemic major decisions had to be made, often on the basis of inadequate scientific data. These data may have been derived from experiments using small sample numbers. Here we review some examples of where this has happened, sometimes with a beneficial outcome and sometimes with a misleading outcome. The identification of BSE as a new disease depended on precise neuropathological observation of a small number of cases rather than the obvious occurrence of large numbers of sick animals. Similarly, the recognition that BSE may have led to disease in humans was based on the neuropathological and clinical picture of new variant Creutzfeldt-Jakob disease (CJD) rather than on an increase in the number of cases of CJD in the UK. Early in the BSE epidemic the possibility that disease could be maternally transmitted from cow to calf was raised, mainly because of a belief that such transmission occurs in scrapie disease of sheep. But, we argue, the evidence for maternal transmission of scrapie, collected in the 1960s, was based on small numbers and is inadequate. Subsequent research has shown a very substantial genetic component in scrapie and epidemiological data show no excess risk in the offspring of affected ewes relative to the risk in the offspring of affected rams. An experiment to determine whether maternal transmission occurs in BSE was flawed and was unable to distinguish between maternal transmission and genetic susceptibility to environmental contamination. An assessment of the risk of BSE to humans depends on determining the levels of infectivity in tissues and transmissibility across species. Data on both of these are deficient, so it is not possible to predict how many people in the UK or elsewhere will become affected with new variant CJD in the next fifty years. The assessment of whether BSE could be transmitted to sheep and whether sheep therefore pose a risk to humans is hampered by a serious lack of evidence about the epidemiology of scrapie in the UK and elsewhere. The UK has paid a heavy price for the BSE epidemic but lessons should be learned from the experience. Every country should have a Specified Offals Ban even if it has no cases of BSE because, by the time it has, it will be too late. Furthermore, the occasional case of BSE should not be regarded as insignificant since it may be the harbinger of an epidemic in the making.     

谷氨酰胺和吉他霉素对早期断奶仔猪血细胞的影响

规模化养猪中重要的环节是仔猪早期断奶,它可同时提高母猪和仔猪的生产力。实践中,仔猪断奶时间由60日龄提前到28日龄或更短。但断奶日龄减少,仔猪应激加大,造成腹泻、水肿和内毒素休克,即“仔猪早期断奶综合征”,严重影响了养猪业的经济效益[1]。目前“早期断奶综合征”的防治常采用添加抗生素等药物手段,虽然抗生素在保证动物的健康状态,限制病原微生物的传播,保持良好的生产性能有一定作用。但使用抗生素所产生的交叉耐药性和药物残留等问题难以解决。因此,寻求一种无残留、降低应激、提高动物抵抗力和生产性能的营养素,是近些年营养学家…     

Biomicroscopy of the tear film: the tear film of the pekingese dog

Polarised light biomicroscopy was used to examine the normal pre-corneal tear film in 21 eyes of 12 pekingese dogs. The purpose of the study was to examine the influence of excessive exophthalmos on the pre-corneal tear film in the dog. The majority of the animals were found to have high levels of ocular surface contamination by particulate material and plaques of viscous mucus. Other abnormalities included surface lipid with an abnormal granular (three dogs) or 'curdled' (two dogs) appearance; excessive thinning of the lipid layer of the tear film; and the presence of dark globular structures in two dogs, which were presumed to be abnormal meibomian lipid. Break up of the tear film was observed in one dog. Grossly, a thread of viscous mucus was frequently observed along the margin of the lower eyelid. It is postulated that this thread forms because of the excessively exophthalmic conformation of the breed, which prevents the normal access of effete mucus and entrapped debris to the lower conjunctival fornix. The combination of the above factors in the pekingese is suggested as the mechanism whereby the tear film has a reduced stability, thus enhancing the risk from factors more usually considered to initiate corneal ulceration in the breed. The possible adverse effects of lid splitting for the mass removal of distichiae in exophthalmic dogs is discussed.