Hematology and chemistry reference values for free-ranging harbor seals (Phoca vitulina) and the effects of hemolysis on chemistry values of captive harbor seals.

Most reported laboratory reference values for harbor seals (Phoca vitulina) are derived from captive seals, or stranded seals that have recovered from disease in marine mammal centers. This study established hematology and serum chemistry reference values for free-ranging harbor seals, using methods and that are current and readily available, and determined the effects of hemolysis on serum chemistry values of captive harbor seals. Blood samples were collected for hematologic and serum chemistry measurements from 14 clinically normal, adult male and female harbor seals and two juvenile harbor seals (approximate age 6 mo) captured in saltwater sloughs and estuaries near Moss Landing, California, USA. Values for amylase, globulin, and differential leukocyte count, not previously reported, were determined. In general, hematology and chemistry values in adults were similar to those reported for free-ranging and captive harbor seals, except for glucose, urea nitrogen, and lactate dehydrogenase (LDH) values, which were higher than those reported previously. Red blood cell counts in the two juveniles were higher than in adults and in young harbor seals studied previously. To determine the effects of hemolysis on serum chemistry values, two intensities of hemolysis were generated experimentally in blood collected from 11 harbor seals recovering from injuries or stranding at the Marine Mammal Center (Sausalito, California 94965, USA). Moderate hemolysis (++, 1 g/L hemoglobin, red-tinged) significantly increased LDH activity, whereas severe hemolysis ( , 2 g/L hemoglobin, cherry red) significantly increased total protein, albumin, calculated globulin, LDH, and total bilirubin and significantly decreased creatinine. The effects of hemolysis must be considered when chemistry results of harbor seals are interpreted.     

Haematology, blood chemistry and urine parameters of free-ranging plains viscachas (Lagostomus maximus) in Argentina determined by use of a portable blood analyser (i-STAT) and conventional laboratory methods

Haematology, blood chemistry and urine values were determined for 44 adult free-ranging plains viscachas (Lagostomus maximus; Rodentia, Chinchillidae) in their pampas habitat in central Argentina. The study animals were captured in the wild and anaesthetized with a ketamine-xylazine combination for physical examination and sampling. Blood was obtained by venipuncture of the saphenous vein. Results for many of the blood parameters fall within the reference ranges for pet chinchillas. Differentiation of white blood cells revealed a predominantly neutrophil count for plains viscachas, while chinchillas have predominantly lymphocytes. Mean values for blood urea nitrogen, creatinine, aspartate aminotransferase and sodium were higher than the upper limit of the reference range for pet chinchillas. The results of seven analytes (haematocrit, haemoglobin, glucose, blood urea nitrogen, sodium, potassium, chloride) were compared by using both a portable blood analyser (i-STAT) in the field and conventional laboratory methods. In general, correlation and agreement between the two methods were low for most parameters.     

Evaluation of a novel haematology analyser for use with feline blood

A novel haematology analyser was evaluated for its use with feline samples. Complete blood cell counts, a five-part differential count, and reticulocyte numbers were determined, and the results compared with reference data. Coefficients of correlation, Passing–Bablok regression analysis and Bland–Altmann difference plots with biases and 95% limits of agreement are reported. Precision and linearity were also studied. The instrument demonstrated very low imprecision, and the tested range of linearity exceeded the reference ranges provided by the manufacturer. For all parameters except monocytes (r = 0.65), the analyser results correlated well with reference methods. Compared with the manual count of aggregated reticulocytes, the instrument showed good agreement with a positive bias. The optical platelet count correlated well with the manual chamber count. In conclusion the analyser was found to be highly reliable for the analysis of feline blood samples in a large veterinary laboratory.     

CLINICAL VALUE OF FRUCTOSAMINE MEASUREMENTS AND FRUCTOSAMINE-ALBUMIN RATIO IN HYPOGLYCEMIC FERRETS (MUSTELA PUTORIUS FURO)

The objective of this preliminary study was to establish a reference range for plasma fructosamine concentration and fructosamine-albumin ratio in healthy ferrets and to compare these reference intervals to values obtained from hypoglycemic ferrets. Fructosamine concentration has been shown to reflect blood glucose concentration over the previous 1 to 2 weeks in other animal species, and may be a useful indicator of chronic hypoglycemia in ferrets diagnosed with insulinoma. Plasma fructosamine was measured with an automated colorimetric assay using nitroblue tetrazolium. Thirty-two clinically healthy and 5 hypoglycemic ferrets were included in the study. The reference interval in healthy ferrets for fructosamine was 110 (98 to 123) – 203 (191 to 218) μmol/L, and the reference interval for plasma fructosamine-albumin ratio was 5.1 (4.6 to 5.6) – 8.6 (8.2 to 9.0) μmol/g. Results for hypoglycemic ferrets were within the ranges for both fructosamine and fructosamine-albumin ratio. As there were no significant differences between the healthy and hypoglycemic ferrets, this study suggests that fructosamine concentration and fructosamine-albumin ratio are not likely to be useful in determining insulinoma-associated chronic hypoglycemia in ferrets.     

Reference intervals for biochemical and haematological parameters in mature domestic donkeys (Equus asinus) in the UK

Reference intervals (RIs) for haematology and serum biochemistry for donkeys in a temperate climate have been previously published using blood sample results from the resident population of a large donkey shelter in the UK. Periodic review of reference intervals is recommended to ensure their applicability to the patient population and changes in laboratory methods and technologies. The current study aimed to revise the previously published haematology and serum biochemistry values for the adult domestic donkey (Equus asinus) in the UK in the light of a change in analytical equipment at the Donkey Sanctuary laboratory, but also to refine the demography of the sample population with respect to age, physiology and clinical history. Clinical pathology results from 138 clinically healthy mature (4–24 years inclusive) female and castrated male donkeys selected from the resident population of the Donkey Sanctuary, were analysed retrospectively. The animals were blood sampled during the period February 2008 to June 2011 as part of a routine health screen prior to rehoming. Results for a total of 38 biochemical and haematological parameters were analysed including 3 previously unreferenced parameters in addition to those assessed in the previous study. The new reference intervals and median values show very poor transferability with recently derived reference intervals for non‐Thoroughbred horses and only limited transferability with reference intervals previously published for donkeys in the UK. Of particular note is a marked reduction in the upper reference limit for triglycerides of 2.8 mmol/l (from 4.3 mmol/l) since this parameter is used to decide when donkeys are at risk of developing hyperlipaemia. This study demonstrates the value of intermittent review of reference intervals and refinement of study populations. Notwithstanding the caution with which reference interval data from different laboratories should be compared, the lack of transferability of results between donkeys and horses highlights the importance of use of species‐appropriate reference intervals for clinical decision‐making.     

The interpretation of serum biochemistry test results in domestic animals

The interpretation of serum biochemistry tests used in the investigation of disease was reviewed. Different methods of calculating reference ranges for biochemical constituents of serum were discussed. Reference ranges in current laboratory use were analyzed statistically to produce means and coefficients of variation.     

Meloxicam pharmacokinetics using nonlinear mixed‐effects modeling in ferrets after single subcutaneous administration

This study was designed to investigate the pharmacokinetics of meloxicam, an oxicam class, nonsteroidal anti‐inflammatory drug (NSAID), in ferrets. We determined the pharmacokinetic properties of a single subcutaneous dose of meloxicam (0.2 mg/kg) in nine male and nine female ferrets. Blood samples were collected by venipuncture of the cranial vena cava into heparinized syringes. Plasma meloxicam concentrations were determined by high‐pressure liquid chromatography (HPLC). Pharmacokinetic variables were calculated using nonlinear mixed‐effects modeling to take advantage of the population‐based sampling scheme and to minimize sample volume collected per animal. Maximum plasma concentration, volume of distribution per absorption, and elimination half‐life were 0.663 μg/mL, 0.21 L, and 11.4 h, respectively, for females and 0.920 μg/mL, 0.35 L, and 17.8 h, respectively, for males. Significant differences were found in each of the above parameters between male and female ferrets. Systemic clearance per absorption was not affected by gender and was 13.4 mL/h. Analgesic efficacy was not evaluated, but plasma meloxicam concentrations achieved in these animals are considered effective in other species. Sex differences in the pharmacokinetic behavior of meloxicam should be taken into consideration when treating ferrets.     

Reference values of chemical blood parameters in puppies during the first eight weeks of life

Reference values were determined for blood plasma concentrations of albumin, total protein, glucose, total bilirubin, urea, creatinine, cholesterin, triglycerides, total calcium und anorganic phosphate as well as the activities of alkaline phosphatase, alanine-amino-transferase (ALT) and glutamate-dehydrogenase based on 109-124 healthy puppies at different times (1st-3rd, 8th-10th, 28th-33rd, and 50th-58th day of life). In addition, all the results were calculated separately for the breeds involved in this study (Beagle [n = 34-40], German Shepherd [n = 32-35] and Golden Retriever [n = 43-53]). Furthermore, male and female puppies were compared. All examined parameters showed remarkable dynamics during the suckling period. Often the values exceeded respectively fell below the reference ranges for adult dogs. Significant systematic breed influences as for instance a significantly lower glucose concentration for the German Shepherd puppies were also found for other parameters, especially for alkaline phosphatase and ALT. However, differences between male and female animals were only present sporadically and were of minor clinical relevance. The gathered breed differences show that it is worthwhile not only to acquire age specific reference ranges but also breed specific reference ranges for selected parameters. On the other hand, reference ranges without the definition of the breed can only be used for orientation.     

Transmission of Mycobacterium bovis from experimentally infected ferrets to non-infected ferrets (Mustela furo)

AIMS: To demonstrate the transmission of Mycobacterium bovis infection from experimentally infected ferrets (Mustela furo) to non-infected ferrets in a laboratory setting, using three different isotypes of M. bovis, and to observe ferret behaviour that might be implicated in disease transmission. METHODS: Three female ferrets, each experimentally infected with a unique strain of M. bovis, were housed together with six female and two male non-infected ferrets in an isolation facility. Transmission of infection was monitored clinically, serologically (using an ELISA test), bacteriologically, histologically, and by isotype analysis of M. bovis isolates using spoligotyping to determine whether or not transmission of each strain occurred. Ferret behaviour was observed using a time-lapse video recorder. RESULTS: Transmission of M. bovis infection was confirmed in two male and four female ferrets. Isotype analysis showed that of the experimentally infected females, one did not infect any other ferret, another transmitted M. bovis to one ferret before it died prematurely 49 days post-infection, and the third, which was cannibalised, appears to have transmitted M. bovis to both males and three females. However, two of these latter three females had died before the event of cannibalism took place. One female was infected with two strains. Several behavioural interactions were observed that could have resulted in M. bovis transmission, including den sharing, sniffing of orifices and faeces, cannibalism and aggressive breeding behaviour. CONCLUSIONS: Horizontal transmission of M. bovis infection was demonstrated in ferrets under experimental housing conditions. Routes of transmission may involve cannibalism and factors such as den sharing, playing, fighting, mating, and sniffing of faeces.     

Serum biochemical and hematologic values of normal and Mycobacterium bovis-infected American bison

Hematologic and serum biochemical tests were used to monitor the health of 3 groups of bison in an experimental study of tuberculosis. Bison were randomly assigned to Mycobacterium bovis-infected, M. bovis-sensitized, and uninfected control groups. Hematologic measurements included total and differential leukocyte counts, hemoglobin (Hb), packed cell volume (PCV), fibrinogen, and plasma proteins. Biochemical tests included serum urea nitrogen, creatinine, aspartate amino transferase, sorbitol dehydrogenase, serum calcium, and serum phosphorus. There were no significant differences (P = 0.05) in any test values between groups of bison. The bison data were combined and compared to similar data of cattle. The mean values for PCV and Hb were higher than values (PCV 24-46%, Hb 8-15 g/dl) for cattle. Mycobacterium bovis-infected bison had a slight increase in the number of blood monocytes and lymphocytes when compared to the uninfected bison but were within the normal ranges for bison and cattle. Other hematologic parameters were within normal ranges reported for cattle. Creatinine levels in all bison were above the normal range (1.0-1.5 mg/dl) for cattle. Phosphorus levels for M. bovis-infected and M. bovis-sensitized bison exceeded the normal range (5.6-8.0 mg/dl) reported for cattle. The level for uninfected bison was near the upper limit of normal for cattle. Mean values for other serum biochemical tests were within the normal ranges reported for cattle.     

Echocardiographic measurements in clinically healthy ferrets anesthetized with isoflurane

Two-dimensional, M4-mode, and color flow Doppler echocardiography was performed in 29 (18 females, 11 males) clinically healthy ferrets anesthetized with isoflurane. M-mode measurements of the left ventricle, left atrial appendage diameter (LAAD), and aorta (Ao) were obtained. The fractional shortening and LAAD/Ao ratio were calculated. The values of the M-mode measurements were compared between the male and female ferrets using a Student's t-test. No significant differences were found. The difference in body weight between the male and female ferrets was highly significant (P<0.001), but no significant correlation was found between body weight and M-mode measurements. Color flow Doppler examinations of the mitral, tricuspid, aortic, and pulmonary valves were recorded and there was minor valvular regurgitation in five ferrets, which was considered nonsignificant.     

I plasma concentrations of adrenocorticotrophic hormone and alpha-melanocyte-stimulating hormone in ferrets (Mustela putorius furo) with hyperadrenocorticism

OBJECTIVE: To determine plasma concentrations of adrenocorticotrophic hormone (ACTH) and alpha-melanocyte stimulating-hormone (alpha-MSH) in healthy ferrets and ferrets with hyperadrenocorticism. ANIMALS: 16 healthy, neutered, privately owned ferrets, 28 healthy laboratory ferrets (21 sexually intact and 7 neutered), and 28 ferrets with hyperadrenocorticism. PROCEDURES: Healthy ferrets were used for determination of reference plasma concentrations of ACTH and a-MSH. Diagnosis of hyperadrenocorticism was made on the basis of history, clinical signs, urinary corticoid-to-creatinine ratios, ultrasonography of the adrenal glands, and macroscopic or microscopic evaluation of the adrenal glands. Blood samples were collected during isoflurane anesthesia. Plasma concentrations of ACTH and alpha-MSH were measured by radioimmunoassay. RESULTS: Plasma concentrations of ACTH in 23 healthy neutered ferrets during the breeding season ranged from 4 to 145 ng/L (median, 50 ng/L). Plasma concentrations of alpha-MSH in 44 healthy neutered or sexually intact ferrets during the breeding season ranged from < 5 to 617 ng/L (median, 37 ng/L). Reference values (the central 95% of the values) for ACTH and alpha-MSH were 13 to 100 ng/L and 8 to 180 ng/L, respectively. Plasma concentrations of ACTH and alpha-MSH in ferrets with hyperadrenocorticism ranged from 1 to 265 ng/L (median, 45 ng/L) and 10 to 148 ng/L (median, 46 ng/L), respectively. These values were not significantly different from those of healthy ferrets. Plasma ACTH concentrations of sexually intact female ferrets in estrus were significantly higher than those of neutered females. CONCLUSIONS AND CLINICAL RELEVANCE: Ferrets with hyperadrenocorticism did not have detectable abnormalities in plasma concentrations of ACTH or alpha-MSH. The findings suggest that hyperadrenocorticism in ferrets is an ACTH and alpha-MSH-independent condition.     

实验用猕猴某些生理生化指标的测定

应用全自动血液细胞分析仪、全自动血液生化分析仪和流式细胞仪等仪器测定和分析人工饲养条件下24只健康猕猴清醒状态下血液一般生理生化指标、CD3 、CD4 、CD8 、CD20 细胞阳性率。结果雌性猕猴平均动脉压、收缩压、舒张压极显著低于雄性猕猴(P<0.01),心电图QRS波、T波差异极显著(P<0.01),Q-T段差异显著(P<0.05)。血液常规指标中雄性猕猴白细胞计数极显著低于雌性猕猴(P<0.01)。猕猴碱性磷酸酶、总胆红素差异极显著(P<0.01)、总胆固醇、γ-谷氨酰转移酶差异显著(P<0.05)、甘油三酯显著低于雄性猕猴(P<0.01),而丙氨酸氨基转换酶,天门冬氨酸氨基转换酶差异极显著(P<0.01)、血清肌酐显著高于雄性猕猴(P<0.05)。CD3 、CD4 、CD8 、CD20 细胞阳性率无显著差别。文章为以猕猴作为实验动物的医学实验提供可靠的参考数据,同时证明一些指标受性别影响,在试验中应受到重视。     

Evaluation of a point-of-care blood analyzer and determination of reference ranges for blood parameters in rockfish

OBJECTIVE: To compare values of blood parameters in rockfish obtained by use of a point-of-care portable blood analyzer with values determined by a veterinary diagnostic laboratory, calculate reference ranges for various blood parameters in black rockfish, and compare values of blood parameters in clinically normal fish with those of fish with clinical abnormalities. DESIGN: Prospective study. ANIMALS: 41 captive adult black rockfish (Sebastes melanops) and 4 captive adult blue rockfish (Sebastes mystinus). PROCEDURE: Rockfish were anesthetized with tricaine methanesulfonate for collection of blood samples. Heparinized blood samples were immediately analyzed with a point-of-care analyzer. Blood sodium, potassium, chloride, urea nitrogen, and glucose concentrations; Hct; pH; partial pressure of carbon dioxide; total carbon dioxide concentration; bicarbonate concentration; base excess; and hemoglobin concentration were determined. A microhematocrit technique was used to determine PCV, and a refractometer was used to estimate total plasma protein concentration. Paired heparinized blood samples were transported to a veterinary diagnostic laboratory for analyses. RESULTS: Data obtained with the point-of-care analyzer were reproducible; however, values for most blood parameters were significantly different from those obtained by the veterinary diagnostic laboratory. Fish with poor body condition had several blood parameter values that were lower than corresponding values in clinically normal fish. CONCLUSIONS AND CLINICAL RELEVANCE: Point-of-care blood analyses may prove useful in rockfish. Point-of-care data for a large number of clinically normal fish must be obtained for reference ranges to be calculated, and further assessments of clinically abnormal fish are necessary to determine the relevance of the data.     

Avian clinical pathology. General considerations

Summary

General aspects of avian clinical pathology are reviewed. It is concluded that in a clinical setting a volume of blood equivalent to 1 per cent of body weight can be collected safely from avian species for laboratory examinations. The anticoagulant of choice for most laboratory investigations is lithium heparin. In most bird species the right jugular vein is the preferred site for routine blood sampling. The use of a vacuum system greatly facilitates the procedure. The importance of immediately processing blood samples is explained. The many variables that may influence haematological or biochemical parameters are discussed and the reasons for determining blood chemical reference values by non‐parametric methods are emphasised.     

Blood biochemical reference ranges for sows under modern management conditions

Published reference ranges for blood biochemistry in swine generally do not relate to sows in modern breeding units, and results were often obtained by methods that are now outdated. The ranges widely used in clinical practice reflect these inappropriate sources. The data presented here were obtained using modern methods of analysis on blood samples from healthy, conventionally managed sows from six breeding herds of known disease status in eastern England, and thus represent appropriate ranges for this class of swine. The values differ from earlier reports principally in higher values for total bilirubin, creatine kinase, and more particularly of total plasma and serum proteins. The latter are shown to be due to higher immunoglobulin concentrations than those previously reported.     

Tuberculosis seroprevalence and comparison of hematology and biochemistry parameters between seropositive and seronegative captive Asian elephants of Nepal

We conducted a tuberculosis (TB) serosurveillance program of captive elephants in Nepal and compared hematology and biochemistry parameters between seropositive and seronegative elephants. A total of 153 elephants (male=20, female=133) from four national parks were tested for TB using the ElephantTB STAT-PAK^® Assay (ChemBio Diagnostic Systems, Inc., Medford, NY, USA). The mean reported age for 138 elephants was 38.5 years (range 2–71 years). Seroprevalence for TB was 21.56% (33/153). The majority of seropositive elephants were female (n=30) and from Chitwan National Park (n=29). The occurrence of TB seropositive cases in other more remote national parks suggests TB may be widespread among the captive elephant population of Nepal. Hematology and biochemistry analyses were performed on 13 and 22 seropositive elephants, respectively and, nine elephants from a seronegative TB herd for comparison. Hematology parameters (hemoglobin, packed cell volume, platelet, white blood cells, and erythrocyte sedimentation rate) were comparable between the two groups. Total protein, globulin, and lactate dehydrogenase were significantly higher in seronegative elephants, and bilirubin was significantly higher in seropositive elephants whereas blood urea nitrogen, creatinine, glutamic oxaloacetic transaminase/aspartate aminotransferase (GOT/AST), glutamic pyruvic transaminase/alanine aminotransferase (GPT/ALT), gamma glutamyl transferase (GT), and albumin were not significantly different. The range of biochemical parameters that were significantly different between seropositive and seronegative elephants had narrow ranges. Thus, the potential of these parameters as a direct biomarker for TB diagnosis is limited based on the findings in this study. We recommend including blood parameters in future TB surveillance studies.     

Composition of cerebrospinal fluid in clinically normal adult ferrets

OBJECTIVE: To determine the protein and cellular composition of CSF in healthy adult ferrets. ANIMALS: 42 clinically normal adult ferrets. PROCEDURE: CSF samples were collected from the cerebellomedullary cistern of anesthetized ferrets by use of disposable 25-gauge, 1.6-cm-long hypodermic needles. Samples were processed within 20 minutes after collection. The number of WBCs and RBCs per microliter of CSF was counted by use of a hemacytometer. The total protein concentration was determined by use of an automated chemistry analyzer. RESULTS: Total WBC counts (range, 0 to 8 cells/microL; mean, 1.59 cells/microL) in CSF of ferrets were similar to reference range values obtained for CSF from other species. Twenty-seven CSF samples had <100 RBCs/microL (mean, 20.3 RBCs/microL). A small but significant effect of blood contamination on WBC counts was found between the 27 CSF samples with <100 RBCs/microL and the remaining samples. Protein concentrations in CSF of ferrets (range, 28.0 to 68.0 mg/dL; mean, 31.4 mg/dL) were higher than has been reported for the CSF of dogs and cats. A significant effect of blood contamination on the CSF protein concentration was not found. CONCLUSION AND CLINICAL RELEVANCE: We have established reference range values for WBC counts and protein concentrations in CSF from healthy adult ferrets that may be useful in the clinical investigation of CNS disease. Results of our study indicate that the WBC count is significantly affected by blood contamination of the CSF sample.     

Effect of temperature,storage time,and sample type on sorbitol dehydrogenase activity in llama serum and plasma

Abstract: Serum and heparinized plasma samples were collected from 11 adult, clinically healthy llamas. Aliquots were assayed for sorbitol dehydrogenase (SDH) activity after storage at room temperature (20°C), 4°C, or −20°C for defined time intervals up to 1 week postcollection. Sorbitol dehydrogenase activity in all samples was within reference intervals for our laboratory. No difference was found between serum and plasma SDH activity when measured immediately (within 1 hour) after collection. Sorbitol dehydrogenase activity decreased to 79% of initial activity by 24 hours in serum stored at room temperature; plasma had 94% of initial SDH activity under the same conditions. Sorbitol dehydrogenase activity was stable in both plasma and serum stored for up to 1 week at 4°C or −20°C. With the exception of serum stored at 20°C for > 8 hours, in vitro stability of llama SDH was adequate for its use in diagnostic testing.