Automated flow cytometric cell count and differentiation of canine cerebrospinal fluid cells using the ADVIA 2120

Background: Microscopic cell counts in cerebrospinal fluid (CSF) are time-consuming and prone to imprecision. The recently introduced automated hematology analyzer ADVIA 2120 offers an automated cell count and differential for CSF in the veterinary software mode based on laser light scatter and absorbance measurements. Objectives: The purpose of this study was to evaluate the precision, linearity, and accuracy of the ADVIA 2120 CSF assay. Methods: Sixty-seven CSF samples were analyzed on the ADVIA 2120 and total nucleated cell counts (TNCC) and RBC counts were compared with the hemocytometer results. In 21 samples with TNCC >5/muL, ADVIA 2120 results were compared with 100-300 cell manual differentials performed on cytocentrifuged preparations. Statistical analysis included Spearman's rank correlation, Passing-Bablok regression, and Bland-Altman analysis. Results: Repeatability (intra-assay) coefficients of variation (CVs) ranged from 4.19% to 25.94%. Interassay CVs ranged from 2.56% to 28.67%. Accurate results within 30% were achieved for TNCC up to 4000/muL. Except for low TNCC, deviation from the expected value was higher (TNCC of 8/muL instead of 4/muL). The following correlation coefficients (r) and biases were achieved compared with the reference method: r=.90 and bias 2.3/muL for TNCC; r=.88 and bias 32.0/muL for RBC counts; r=.86 and bias +/-13.4% for mononuclear and polymorphonuclear cell percentages; r=.88 and bias -6.1% for lymphocyte percentage; r=.56 and bias 19.4% for monocyte percentage; and r=.75 and bias -9.7% for neutrophil percentage. Conclusion: Our results demonstrated that the automated ADVIA 2120 CSF assay generally compares well with reference methods although there are some limitations for the automated monocyte count and for samples with only mild pleocytosis.     

Peritoneal fluid analysis in adult, nonpregnant Awassi sheep

BACKGROUND: To the authors' knowledge, there is no information in the literature about normal peritoneal fluid values in ovine species. OBJECTIVES: The purpose of the study reported here was to establish reference intervals for peritoneal fluid from clinically normal Awassi sheep and to compare the values to those in blood. METHODS: Peritoneal fluid and blood samples were collected into tubes containing EDTA, from 40 clinically healthy, nonpregnant, female Awassi sheep, aged 2 to 7 years. Total nucleated cell count (TNCC) was determined using an electronic cell counter. Total protein, albumin, urea, creatinine, and glucose concentrations and aspartate transaminase activity were analyzed using commercially available kits. RESULTS: TNCC (mean +/- SD) of peritoneal fluid was 1.1 +/- 0.87 X 10(3)/microl, with neutrophils (3.9%), lymphocytes (33.5%), macrophages/monocytes (61.2%), and eosinophils (1.4%). Biochemical results in peritoneal fluid were: total protein, 1.7 +/- 0.74 g/dL; albumin, 1.0 +/- 0.04 g/dL; urea, 12.6 +/- 3.95 mg/dL; creatinine, 0.6 +/- 0.19 mg/dL; glucose, 54.8 +/- 6.11 mg/dL; and aspartate transaminase, 23.5 +/- 8.82 U/L. Eosinophil percentage and creatinine concentration did not differ significantly from blood values. CONCLUSION: Baseline values for cytologic and biochemical parameters in peritoneal fluid of Awassi sheep, with comparison to blood, have been generated. Such data may be applicable to other ovine species and can be used in the clinical investigation of ovine abdominal disorders.     

Metabolic profiles in goat does in late pregnancy with and without subclinical pregnancy toxemia

Background: Pregnant goat does can develop various metabolic diseases during late pregnancy that may have profound effects on their health and productivity, including subclinical pregnancy toxemia. Objectives: This study was performed to evaluate serum biochemical findings in goats in late pregnancy with and without subclinical pregnancy toxemia. Methods: Serum samples were obtained from 153 clinically healthy goats in the last trimester of pregnancy from 14 herds in central and northern Jordan. Serum was analyzed for the following constituents: 3‐hydroxybutyrate (3‐HB), glucose, total protein, albumin, urea, creatinine, aspartate aminotransferase, alkaline phosphatase, cholesterol, triglycerides, cortisol, T3, T4, total calcium, magnesium, and phosphorus, using commercially available kits. Goats were classified as having subclinical pregnancy toxemia or as controls on the basis of 3‐HB results. Biochemical results were compared between the 2 groups using nonparametric statistical tests. Results: Fifty‐three does had subclinical pregnancy toxemia (3‐HB>0.86 mmol/L) and 100 were pregnant control does (3‐HB≤0.86 mmol/L). Does with subclinical pregnancy toxemia had significantly lower glucose concentrations (P<.001) and significantly higher urea (P=.042) and total protein (P=.048) concentrations compared with pregnant control does. 3‐HB concentration was significantly correlated with glucose (r=0.290, P=.035), phosphorus (r=0.351, P=.01), and cortisol (r=0.394, P=.004) concentrations in goats with subclinical toxemia. Conclusions: Results of this study indicate that the biochemical profile of does with subclinical pregnancy toxemia differs from that of other pregnant goats. 3‐HB should be measured in pregnant does in late stages of gestation with hypoglycemia or with elevated urea and total protein concentrations.     

Cytologic interpretation of canine cerebrospinal fluid samples with low total nucleated cell concentration,with and without blood contamination

Background: Cerebrospinal fluid (CSF) is potentially altered by iatrogenic blood contamination at the time of sampling due to the addition of blood‐associated leukocytes and protein. Objectives: The objective of this study was to assess whether protein concentration, neutrophil percentage, and the presence of activated macrophages, reactive lymphocytes, or eosinophils in CSF samples with low total nucleated cell concentration (TNCC) are affected by blood contamination or associated with central nervous system (CNS) disease. Methods: Case records from the Royal Veterinary College Diagnostic Laboratory were searched retrospectively for dogs with CSF having ≤5 TNCC/μL. TNCC, RBC, and protein concentrations; neutrophil percentage; and the presence of activated macrophages, reactive lymphocytes, and eosinophils were recorded. Results of magnetic resonance imaging (MRI) also were recorded as a marker of CNS disease. Results: Of 906 cases evaluated, 106 (12%) had blood contamination (>500 RBCs/μL) in CSF. Protein concentration and neutrophil percentage were significantly higher and the presence of eosinophils was more likely in blood contaminated vs noncontaminated samples. Non‐blood‐contaminated samples with activated macrophages or reactive lymphocytes had higher protein concentrations and neutrophil percentages, and those with activated macrophages were more likely to have a positive finding on MRI. Conclusions: Protein concentration, neutrophil percentage, and the presence of eosinophils are significantly affected by blood contamination in canine CSF having low TNCC. Activated macrophages and reactive lymphocytes are not affected by blood contamination, however, and may be useful in identifying dogs with CNS abnormalities.     

Canine complete blood counts: a comparison of four in‐office instruments with the ADVIA 120 and manual differential counts

Background: With more use of bench‐top in‐office hematology analyzers, the accuracy of reported values is increasingly important. Instruments use varied methods for cell counting and differentiation, and blood smears may not always be examined. Objective: The purpose of this study was to compare canine CBC results using 4 bench‐top instruments (Hemavet 950, Heska CBC‐Diff, IDEXX LaserCyte, and IDEXX VetAutoread) with ADVIA 120 and manual leukocyte counts. Methods: EDTA‐anticoagulated canine blood samples (n=100) were analyzed on each instrument. Manual differentials were based on 100‐cell counts. Linear regression, difference plots, paired t‐tests, and estimation of diagnostic equivalence were used to analyze results. Results: Correlations of HCT, WBC, and platelet counts were very good to excellent between all in‐office instruments and the ADVIA 120, but results varied in accuracy (comparability). Hemavet 950 and Heska CBC‐Diff results compared best with ADVIA results and manual leukocyte differentials. HCT and platelet counts on the IDEXX VetAutoread compared well with those from the ADVIA. Except for neutrophil counts, leukocyte differentials from all instruments compared poorly with ADVIA and manual counts. Reticulocyte counts on the LaserCyte and VetAutoread compared poorly with those from the ADVIA. Conclusions: The Hemavet 950 and Heska CBC‐Diff performed best of the 4 analyzers we compared. HCT, WBC, and platelet counts on the LaserCyte had minimally sufficient comparability for diagnostic use. Except for neutrophils (granulocytes), leukocyte differential counts were unreliable on all in‐office analyzers. Instruments with a 5‐part leukocyte differential provided no added benefit over a 3‐part differential. Assessment of erythrocyte regeneration on the LaserCyte and VetAutoread was unreliable compared with the ADVIA 120.     

Prospective study of blackthorn injury and synovitis in 35 horses

The objective of this prospective clinical study was to investigate the cause and describe the presentation, diagnosis, treatment techniques and outcome of Prunus spinosa (blackthorn) injury and synovitis in the horse. In all cases presented with blackthorn injury and synovitis, surgical treatment was performed within 24 h, using a two-stage procedure: 1-Perisynovial technique using ultrasound guided electrosurgical dissection; 2-Endoscopic technique. The diagnosis was confirmed by retrieval of black plant material from or close to the affected synovial structure. Mean lameness score on presentation was 4/5 (range 1–5). The most commonly affected structures were extensor tendon sheaths (12/35) and fetlock joints (11/35). All cases had thorn material removed, 80% had thorn material removed at surgery and in 49% it was intra-synovial. On presentation, the mean synovial fluid total protein level (TP) was 47.6 g/L (range 18–66); mean total nucleated cell count (TNCC) was 176 × 10⁹ cells/L (range 12–312). Two days post-surgery, mean total protein levels were 33 g/L (range 16–52), mean TNCC was 13 × 10⁹ cells/L (range 1–35). At 5 days post-surgery, the mean total protein was 23 g/L (range 12–28) and TNCC was 5 × 10⁹ cells/L (range 1–12). All synovial fluid cultures were negative. Twenty-eight (80%) horses were sound 5 days post-operatively, seven (20%) were not lame in walk; they all returned to full work in an average time of 8 weeks (range 3–48 weeks). Surgery achieved accurate identification and removal of thorn material. In contrast to previous studies of synovial sepsis, these cases had a positive outcome despite high pre- and post-operative synovial fluid TP and TNCC. These findings suggest that Prunus spinosus (blackthorn) synovitis has a different aetiology to synovitis originating from sepsis or other types of contamination.     

Thrombelastography in dogs admitted to an intensive care unit

Background: Underlying conditions in dogs admitted to an intensive care unit (ICU) can cause hemostatic dysfunction. Thrombelastography (TEG) may be useful in detecting hemostatic alterations as compared with standard coagulation tests. Objectives: The purpose of this study was to compare TEG results and those of standard coagulation tests in identifying hemostatic dysfunction in dogs admitted to an ICU and to investigate associations among the variables measured. Methods: Tissue factor‐activated TEG analysis, d ‐dimer and fibrinogen concentrations, antithrombin (AT) activity, prothrombin time (PT), activated partial thromboplastin time (aPTT), and platelet count were measured using standard techniques on 27 dogs admitted to ICU with a disease known to be associated with hemostatic dysfunction and in 31 clinically healthy control dogs. Results were compared between groups using nonparametric tests and κ analysis; principal component analysis (PCA) and Spearman rank correlation were used to measure associations among variables. Results: Fourteen of 27 ICU dogs had abnormal TEG tracings, which were used to classify the dogs as hypercoagulable (n=11), hypocoagulable (n=3), or normocoagulable (n=13). Hypercoagulable dogs had significantly increased d ‐dimer (P=.03) and fibrinogen (P=.01) concentrations compared with normocoagulable dogs. In ICU dogs, positive associations were identified between maximum amplitude (MA), α‐angle, fibrinogen concentration, and platelet count, and between PT, aPTT, and reaction time (R). Significant correlations were found between MA and fibrinogen (r_s=.76, P<.001) and between reaction time (R) and PT (r_s=.51, P=.003). Conclusions: TEG was useful in detecting hemostatic dysfunction in dogs in an ICU. Positive associations among variables may provide insight as to how overall coagulation status reflects alterations in clot strength and coagulation time. Dogs with TEG tracings indicative of hypercoagulability are likely in procoagulant states. Future studies of the incidence of thrombotic complications in dogs with hypercoagulable TEG tracings are warranted.     

Evaluation of serum haptoglobin and C‐reactive protein in dogs with mammary tumors

Background: In veterinary medicine, there is increasing interest in measuring acute phase proteins as a tool in the diagnosis and monitoring of neoplastic diseases. Although mammary neoplasms are the most common type of cancer in dogs, acute phase proteins have not been extensively evaluated in dogs with mammary tumors. Objectives: The aim of this study was to evaluate serum haptoglobin (Hp) and C‐reactive protein (CRP) concentrations in the dogs with mammary tumors and assess their potential association with malignancy. Methods: A retrospective study of dogs with mammary tumors was performed. Serum concentrations of CRP and Hp were determined in healthy control dogs (n=20) and dogs with mammary tumors before surgery (n=41). Mammary tumors were grouped as carcinomas (n=24), fibrosarcoma (n=1), malignant mixed tumors (n=7), benign mixed tumors (n=6), and adenomas (n=3). CRP and Hp concentrations were compared in dogs with different tumor types and were also compared based on tumor size, lymph node infiltration, skin ulceration, fixation to underlying tissue, and time between tumor identification and removal. Results: Hp concentration was significantly (P<.043) higher in dogs with mammary tumors (median 2.03 g/L, range 0.09–2.94 g/L) compared with controls (1.38 g/L, range 0.08–3.00 g/L), but the range of values overlapped considerably. CRP concentration was higher in dogs with carcinomas (4.70 mg/L, range 0.63–128.96 mg/L) vs controls (2.11 mg/L, range 0.25–6.57 mg/L) (P=.0008) and in dogs with ulcerated skin (14.8 mg/L, range 5.7–128.9 mg/L, n=3) compared with those without ulceration (2.4 mg/L, range 0.11–30.3 mg/L, n=38) (P=.048). Conclusions: Serum Hp and CRP do not appear to have value in diagnosing or predicting malignancy of mammary tumors in dogs. Higher CRP concentrations in dogs with mammary carcinoma suggest a role for inflammation in this tumor type.     

Electrocardiographic assessment of hyperkalemia in dogs and cats

Objective: To determine if electrocardiogram (ECG) changes induced by hyperkalemia in clinical patients correspond with previously reported changes in experimental animals. Design: Prospective clinical study. Setting: Two private practice 24‐hour emergency and critical care facilities. Animals: Fifteen dogs and 22 cats with serum potassium levels >5.5 mEq/L. Interventions: None. Measurements: The following data were collected when hyperkalemia was documented: ECG (n=37), sodium and chloride (mEq/L) (n=35), total magnesium (mg/dL) (n=18), total calcium (mg/dL) (n=30), and venous pH (n=18). Animals were divided into five groups based on severity of hyperkalemia and ECG interpretation included rate, rhythm and P‐QRS‐T evaluation. Main Results: Twenty‐two of 37 (59%) of the ECGs were normal or revealed abnormalities that have not been previously described in conjunction with hyperkalemia. In dogs, there was no correlation (r=0) between potassium blood levels and heart rate (n=15). There was weak correlation (r=0.40; P=0.06) between potassium blood levels and heart rate in cats (n=22). The correlation was stronger (r=0.64; P<0.05) when data were compared in cats with serum potassium level >8.5 mEq/L (Groups 4 and Group 5; n=11). Conclusions: ECGs obtained from ill dogs and cats with hyperkalemia are inconsistent with ECGs from experimentally induced hyperkalemia. It is difficult to determine the clinical relevance of heart rate differences between cats with serum potassium levels >8.5 mEq/L and animals with experimentally induced hyperkalemia; this may be due to the presence of other biochemical abnormalities in diseased animals.     

Comparative clinical study of canine and feline total blood cell count results with seven in‐clinic and two commercial laboratory hematology analyzers

Background: A CBC is an integral part of the assessment of health and disease in companion animals. While in the past newer technologies for CBC analysis were limited to large clinical pathology laboratories, several smaller and affordable automated hematology analyzers have been developed for in‐clinic use. Objectives: The purpose of this study was to compare CBC results generated by 7 in‐clinic laser‐ and impedance‐based hematology instruments and 2 commercial laboratory analyzers. Methods: Over a 3‐month period, fresh EDTA‐anticoagulated blood samples from healthy and diseased dogs (n=260) and cats (n=110) were analyzed on the LaserCyte, ForCyte, MS45, Heska CBC, Scil Vet ABC, VetScan HMT, QBC Vet Autoread, CELL‐DYN 3500, and ADVIA 120 analyzers. Results were compared by regression correlation (linear, Deming, Passing‐Bablok) and Bland–Altman bias plots using the ADVIA as the criterion standard for all analytes except HCT, which was compared with manual PCV. Precision, linearity, and carryover also were evaluated. Results: For most analytes, the in‐clinic analyzers and the CELL‐DYN performed similarly and correlated well with the ADVIA. The biases ranged from ?0.6 to 2.4 × 10⁹/L for WBC count, 0 to 0.9 × 10¹²/L for RBC count, ?1.5 to 0.7 g/dL for hemoglobin concentration, ?4.3 to 8.3 fL for MCV, and ?69.3 to 77.2 × 10⁹/L for platelet count. Compared with PCV, the HCT on most analyzers had a bias from 0.1% to 7.2%. Canine reticulocyte counts on the LaserCyte and ForCyte correlated but had a negative bias compared with those on the ADVIA. Precision, linearity, and carryover results were excellent for most analyzers. Conclusions: Total WBC and RBC counts were acceptable on all in‐clinic hematology instruments studied, with limitations for some RBC parameters and platelet counts. Together with evaluation of a blood film, these in‐clinic instruments can provide useful information on canine and feline patients in veterinary practices.     

Effect of delipidant agents during in vitro culture on the development,lipid content,gene expression and cryotolerance of bovine embryos

In vitro produced embryos are still sensitive to the freezing process which can be explained, in part, by the high-lipid accumulation that characterizes these embryos. Therefore, we aimed to evaluate the effect of delipidating agents, L-carnitine and the trans-10 cis-12 conjugated linoleic acid (CLA) isomer, on blastocyst development, lipid content, gene expression and cryotolerance when added to embryo culture media. Embryos were cultured in four different media: T1: control (n = 616), synthetic oviduct fluid (SOF) media with 5% foetal bovine serum (FBS); T2: L-carnitine (n = 648), SOF medium with 5% FBS and 0.6 mg/ml of L-carnitine; T3: CLA (n = 627), SOF medium with 5% FBS and 100 μM trans-10 cis-12 CLA; and T4: L-carnitine + CLA: (n = 597), SOF medium with 5% FBS plus 0.6 mg/ml L-carnitine and 100 μM trans-10 cis-12 CLA. Supplementation of culture medium with either or both delipidating agents reduced (p < .05) blastocyst rate on D7 (T1 = 49 ± 3.5; T2 = 39 ± 3.0; T3 = 42 ± 3.9 and T4 = 39 ± 3.9), but did not affected gene expression (p > .05). Although embryos cultured in the presence of L-carnitine contained fewer (p < .05) lipid droplets than the control embryos, they showed a lower re-expansion rate 24 hr post-thaw than those (p < .05). In conclusion, although L-carnitine reduced the amount of lipids in cultured embryos, the use of L-carnitine and CLA during in vitro culture was not able to improve the embryo production and the response to cryopreservation.     

Neutrophilic myeloperoxidase index and mean light absorbance in neonatal septic and nonseptic foals

Background: Two neutrophilic indices reported by the ADVIA 120 Hematology Analyzer, neutrophilic myeloperoxidase index (MPXI), and mean light absorbance (neutrophil X mean [NXM]) have been proposed as indicators of systemic inflammatory disease in horses and of neutrophil activation in coronary ischemic syndromes in people. Objective: The aim of this study was to evaluate NXM and MPXI in healthy, sick nonseptic, and sick septic foals to determine whether conditions likely associated with neutrophil activation result in decreases in these variables. Methods: In this retrospective study, CBC data from 61 neonatal foals presented to the Equine Teaching Hospital of Barcelona were evaluated for correlations between MPXI, NXM, percentage of large unstained cells, neutrophil count, and percentage of band neutrophils. Results obtained in septic (n=32), sick nonseptic (n=22), and healthy foals (n=7) were compared. In addition, results recorded in septic/neutropenic (n=12), septic/non‐neutropenic (n=20), nonseptic/neutropenic (n=8), nonseptic/non‐neutropenic (n=14), and healthy foals (n=7) were also compared. Results: A weak negative correlation was found between MPXI and neutrophil count and between NXM and percentage of band neutrophils. Septic/neutropenic foals had significantly higher MPXI values (median 17.9, minimum–maximum 4.7–42.5) than did septic/non‐neutropenic (1.5, ?24.4 to 22.3), nonseptic/neutropenic (6.6, 0.6–17.9), and nonseptic/non‐neutropenic foals (8.8, ?10.1 to 16.8) but did not differ significantly from controls (12.8, ?8.5 to 20.4). Conclusions: Significant differences in NXM or MPXI were not found when disease groups were compared with controls; however, septic/neutropenic foals had significantly higher median MPXI than other groups of sick foals. Further prospective studies are needed to clarify if this finding is related to decreased neutrophil function or activation in septic/neutropenic foals.     

Automated counting of nucleated cells in equine synovial fluid without and with hyaluronidase pretreatment

Background: Microscopy is usually used to obtain manual total and differential cell counts in equine synovial fluid. A faster, more precise method is desirable. Objectives: The objectives were to compare an automated impedance method with a manual method for obtaining total and differential cell counts in equine synovial fluid and to evaluate the effect of pretreatment with hyaluronidase on automated results. Methods: Synovial fluid samples (n=48) were collected into EDTA and analyzed within 48 hours. Automated total and differential cell counts were evaluated using a Medonic CA620‐VET hematology analyzer before and after pretreatment for 5–30 minutes with hyaluronidase (final concentration 0.01 mg/mL). A hemacytometer count and microscopic evaluation of a direct smear were used as the reference method. Intra‐assay coefficients of variation (CV) were determined. Results: Thirty‐one of 46 untreated samples and 0/46 hyaluronidase‐treated samples were error‐flagged by the analyzer. Correlation between automated (ANCC) and manual (MNCC) nucleated cell counts in untreated samples (n=15; R²=0.93) and pretreated samples (n=46; R²=0.94) was high, and pseudomedian difference was low. Intra‐assay CVs for samples with medium and high cellularity were significantly lower for ANCC (1.5–2.7%) compared with MNCC (6.1–15.7%) (P<.01). Valid automated differential cell counts were not obtained. Conclusions: Automated total cell counts obtained on the Medonic analyzer correlate well with manual counts in equine synovial fluid; however, pretreatment with hyaluronidase is required to minimize error flags. Automated differential counts are not accurate for synovial fluid.     

Evaluation of peripheral blood neutrophil function in tumor‐bearing dogs

Background: Peripheral blood neutrophils of untreated human cancer patients have been shown to have normal, increased, and decreased phagocytic activity, killing capacity, and/or oxidative burst activities. Objectives: The objectives of this study were to evaluate oxidative burst and phagocytic activities of peripheral blood neutrophils from tumor‐bearing dogs before therapy and compare them with neutrophil function of healthy control dogs. Methods: Heparinized whole blood was obtained from dogs with high‐grade lymphoma (n=23), sarcoma (n=13), or carcinoma (n=11), and healthy control dogs (n=11) for flow cytometric evaluation of oxidative burst and phagocytic activities. Percentage of bursting cells and amount of oxidative burst activity were determined after stimulation with phorbol 12‐myristate 13‐acetate (PMA) or Escherichia coli. Percentage of phagocytic cells and amount of phagocytic activity were determined after incubation with fluorescent E. coli. Results: Compared with control dogs, dogs with sarcoma (P=.004) and carcinoma (P=.05) had a lower percentage of neutrophils exhibiting oxidative burst activity after stimulation with PMA. Phagocytic activity was significantly lower in dogs with sarcomas compared with control dogs (P<.0001) and dogs with lymphoma (P=.01). Conclusions: Untreated carcinomas and sarcomas in dogs may suppress the percentage of neutrophils capable of oxidative burst when stimulated by PMA. Furthermore, sarcomas also may suppress the amount of phagocytic activity per neutrophil. Until further studies can be performed, the clinical significance of these findings is unknown.     

Postpartum follicular development in Brahman cows under two stocking rates

The aim was to study postpartum follicular population dynamics in Brahman cows in the Mexican humid tropics and under different stocking rates. Lactating adult cows (n = 104) grazed native grammas (Paspalum spp., Axonopus spp.) under two stocking rates: 2.8 (high) or 1.7 (low) cows/ha. Follicle numbers and their size (millimetres) were measured by ovarian ultrasound scanning. Animals detected in oestrus and inseminated were taken out of the study. Both stocking rates produced similar (P < 0.05) body condition scores (BCS) and follicular dynamics. The percent of follicles, averaged over stocking rates, of <4 mm diminished and those of 5–8 mm increased with time, whilst those of >9 mm remained constant; respective values for the previously mentioned sizes were 60 days postpartum (n = 101), 49%, 56% and 5%; 120 days (n = 70), 28%, 66% and 6%; and 180 days (n = 15), 9%, 79% and 12%. There was a positive correlation between BCS and follicle number for size <4 mm (r = 0.73, P < 0.01), whilst for size 5–8 mm, this was negative (r = −0.51, P < 0.05), for follicles >9 mm, no difference was found (r = −0.08, P > 0.05). It was concluded that stocking rate did not affect follicular population dynamics of Brahman cows, but BCS dictated the number and categories of follicles present regardless of the stocking rate utilised.     

Using urea dilution to standardise components of pleural and bronchoalveolar lavage fluids in the dog

AIM: To develop a technique to estimate the volume of epithelial lining fluid (ELF) obtained during bronchoalveolar lavage (BAL) and pleural lavage (PL) in the dog, using the urea dilution method.

METHODS: BAL and PL fluids were obtained by saline lavage of pulmonary and pleural cavities of nine clinically healthy mixed-breed dogs immediately after euthanasia. Cell counts in the BAL and PL fluids were measured using standard techniques. The concentration of ELF in each lavage fluid was calculated from the relative concentration of urea in plasma and in each type of lavage fluid. Cell counts in ELF were then calculated.

RESULTS: There were substantially higher cell counts in ELF compared to BAL or PF fluid. However, nucleated cell counts in ELF could not be predicted from cell counts in BAL or PL fluid.

CONCLUSIONS AND CLINICAL RELEVANCE: These results suggest that accurate assessment of cellular or non-cellular components in lavage fluids should include a calculation of the proportion of ELF recovered, using a method such as urea dilution.     

Association of cerebrospinal fluid analysis findings with clinical signs and outcome in acute nonambulatory thoracolumbar disc disease in dogs

Background: Cerebrospinal fluid (CSF) pleocytosis recently was associated with the severity of neurologic signs in dogs with intervertebral disc disease (IVDD). Hypothesis/Objectives: To look for an association among CSF cell counts, total protein concentration, and severity of neurologic signs at presentation with outcome in dogs with acute thoracolumbar IVDD. Our hypothesis was that CSF total nucleated cell count (TNCC) and percentage cell types would be associated with the severity of spinal cord damage and therefore with both the presenting clinical signs and the prognosis of affected dogs. Animals: Fifty‐four dogs with acute nonambulatory thoracolumbar IVDD were evaluated. Methods: Retrospective study. Signalment, neurologic grade, CSF TNCC, protein concentration, red blood cells count and differential cell percentages, and short‐ and long‐term outcomes were evaluated. Results: CSF pleocytosis (>5 cells/μL) was present in 54% of dogs and was positively associated with neurologic grade at presentation and with postoperative time to regaining ambulation. Neutrophils were observed most frequently. The percentage of CSF macrophages and macrophage to monocyte ratio were higher (P= .001, for both) in dogs presented without deep pain sensation (DPS) that did not regain ambulation. Receiver operator characteristics curve analysis yielded a cut‐off point of 13% macrophages with a sensitivity and specificity of 100 and 83%, respectively, for prediction of a negative outcome. Conclusions and Clinical Importance: CSF pleocytosis is positively associated with the severity of spinal cord damage in dogs with thoracolumbar IVDD. The percentage of CSF macrophages can be used as a prognostic indicator for regaining ambulation in dogs that have lost DPS.     

Synovial fluid cell counts and total protein concentration in clinically normal fetlock joints of young dromedarian camels

Twenty-seven 9-12 months old healthy male dromedarian camels were used to determine total nucleated leucocyte count (TNCC), absolute and percentages of polymorphonuclear (PMN) and mononuclear leucocytes, and total protein (TP) concentration in synovial fluid from grossly and radiographically normal fetlock joints. Arthrocentesis was performed bilaterally from the fetlock joints of the forelimbs and hindlimbs. Blood contaminated samples and samples obtained from grossly or radiographically abnormal joints were excluded. The mean +/- SD of TNCC in 108 samples of fetlock joint synovial fluids was 500 +/- 400 cells/microl. Monocytes/macrophages were the predominant cell type. There were no significant differences in mean TNCC, absolute numbers and percentages of various leucocytes and TP concentrations between the right and left fetlock joints of the forelimbs and hindlimbs or between the fetlock joints of the forelimbs and hindlimbs. The mean +/- SD of absolute numbers and percentages of various cell types were: PMN leucocytes 1 +/- 2 cells/microl (2%), lymphocytes 116 +/- 167 cells/microl (26%), and monocytes/macrophages 383 +/- 323 cells/microl (72%). The mean +/- SD of TP concentration was 2 +/- 1 g/dl.     

Disposition of ampicillin trihydrate in plasma,uterine tissue,lochial fluid,and milk of postpartum dairy cattle

The objective of this study was to determine the disposition of ampicillin in plasma, uterine tissue, lochial fluid, and milk of postpartum dairy cattle. Ampicillin trihydrate was administered by intramuscular (i.m.) injection at a dose of 11 mg/kg of body weight every 24 h (n = 6, total of 3 doses) or every 12 h (n = 6, total of 5 doses) for 3 days. Concentrations of ampicillin were measured in plasma, uterine tissue, lochial fluid, and milk using HPLC with ultraviolet absorption. Quantifiable ampicillin concentrations were found in plasma, milk, and lochial fluid of all cattle within 30 min, 4 h, and 4 h of administration of ampicillin trihydrate, respectively. There was no significant effect of dosing interval (every 12 vs. every 24 h) and no significant interactions between dosing interval and sampling site on the pharmacokinetic variable measured or calculated. Median peak ampicillin concentration at steady‐state was significantly higher in lochial fluid (5.27 μg/mL after q 24 h dosing) than other body fluids or tissues and significantly higher in plasma (3.11 μg/mL) compared to milk (0.49 μg/mL) or endometrial tissue (1.55 μg/mL). Ampicillin trihydrate administered once daily by the i.m. route at the label dose of 11 mg/kg of body weight achieves therapeutic concentrations in the milk, lochial fluid, and endometrial tissue of healthy postpartum dairy cattle.     

Direct colloid osmometry in healthy New World camelids

Background: Direct colloid osmometry provides an objective assessment of the oncotic effects of crystalloid or colloidal fluid therapy, which is especially useful in monitoring fluid therapy of critically ill camelids due to their tendency toward nonspecific hypoproteinemia with increased risk of developing edema and ascites. Objectives: The aims of this study were to measure colloid osmotic pressure (COP) of alpacas and llamas, determine its correlation with concentrations of total protein (TP) and total solids (TS), as well as both albumin (A) and globulin (G) concentrations in the same model (A+G), and evaluate the effects of sample type and storage conditions on COP. Methods: Blood was collected from clinically healthy alpacas (n=23) and llamas (n=22) into heparin tubes. COP of fresh whole blood (COP_FB) and plasma (COP_FP) was determined using a membrane osmometer. For 20 alpacas, COP of refrigerated whole blood (COP_RB) and frozen plasma (COP_FrP) was also measured. Correlations between COP_FB and TS, TP, and A+G concentrations were assessed by simple and multiple regression analysis to model potential predictors. Results: Median COP_FB from alpacas (24.6 mmHg, range 19.3–28.1) was not significantly different from that of llamas (25.3 mmHg, range 22.5–33.7). Sample type or storage conditions did not affect COP. Measured COP had a strong positive linear correlation with TS, TP, and A+G concentrations in alpacas (r²=.7, .74, and .88, respectively). In llamas, COP correlated best with TS concentration (r²=.59), whereas correlation with TP and A+G concentrations was poor (r²=.19 and .25, respectively). Conclusion: COP can be measured using heparinized whole blood or plasma, either fresh or stored. Direct measurement is recommended whenever quantitative knowledge of COP is required in clinical or research setting. Further studies are needed to verify if the poor association of COP with TP found in this study can be generalized to llamas.