Evaluation of the QBC-vet autoread haematology system for domestic and pet animal species

A comprehensive evaluation was initiated to determine the suitability of the QBC-Vet Autoread haematology system for veterinary purposes in domestic and pet animal species. The system determines haematocrit (HCT), haemoglobin (HGB), white blood cell (WBC) count, granulocyte count, combined lymphocyte and monocyte count (L/M), platelet count, as well as eosinophil and neutrophil counts (canine samples only), and reticulocyte count (canine and feline samples only). Linearity assessed for a canine sample usually surpassed the physiological range. Within-batch precision was very good for the majority of the parameters in feline and canine samples: Coefficients of variation (CV) were below 5.5% for HCT, HGB and WBC. In order to test the accuracy of the system with respect to reference methods, a total of 300 blood samples from the Clinics of Internal Veterinary Medicine and Surgery (101 canine, 99 feline and 100 equine) were analysed. Strong linear correlation was demonstrated for HCT, HGB (rτ;/0.96) and WBC (rτ;/0.93) based on high correlation coefficients and narrow confidence intervals. A somewhat higher degree of variation from the estimated regression lines was found in differential blood cell counts, especially for eosinophil counts of the dog where the automated reader erroneously attributed some lymphocytes or monocytes to eosinophil counts. Accuracy of the system was also assessed with respect to clinical relevance of results. The majority of leukocytosis (50 of 53), neutrophilias (3 of 4), or eosinophilias (4 of 5) was detected properly by the QBC-Vet Autoread haematology system, but only 20 out of 35 leukopenic samples were identified correctly. The system detected the presence of reticulocytes in the majority of feline (9 of 10) and canine (6 of 7) samples with a regenerative anaemia. Unexpectedly, platelets of cats were measured with high within-batch precision (mean CV=4.64%). No ‘streaming' effect (no discrimination between erythrocytes and granulocytes) was observed with this advanced QBC system. The system was found to be easy both in handling and interpretation of results. The buffy coat profile appeared particularly useful and informative. In conclusion, the QBC-Vet Autoread-System has excellent analytical properties and is well suited for veterinary purpose

Canine Cerebrospinal Fluid Analysis Using Two New Automated Techniques: The Sysmex XN-V Body Fluid Mode and an Artificial-Intelligence-Based Algorithm

Cerebrospinal fluid analysis is an important diagnostic test when assessing a neurological canine patient. For this analysis, the total nucleated cell count and differential cell counts are routinely taken, but both involve time-consuming manual methods. To investigate faster automated methods, in this study, the Sysmex XN-V body fluid mode and the deep-learning-based algorithm generated by the Olympus VS200 slide scanner were compared with the manual methods in 161 canine cerebrospinal fluid samples for the total nucleated cell count and in 65 samples with pleocytosis for the differential counts. Following incorrect gating by the Sysmex body fluid mode, all samples were reanalyzed with manually set gates. The Sysmex body fluid mode then showed a mean bias of 15.19 cells/μL for the total nucleated cell count and mean biases of 4.95% and −4.95% for the two-part differential cell count, while the deep-learning-based algorithm showed mean biases of −7.25%, −0.03% and 7.27% for the lymphocytes, neutrophils and monocytoid cells, respectively. Based on our findings, we propose that the automated Sysmex body fluid mode be used to measure the total nucleated cell count in canine cerebrospinal fluid samples after making adjustments to the predefined settings from the manufacturer. However, the two-part differential count of the Sysmex body fluid mode and the deep-learning-based algorithm require some optimization

Comparison of Sysmex XN-V body fluid mode and deep-learning-based quantification with manual techniques for total nucleated cell count and differential count for equine bronchoalveolar lavage samples

Background: Bronchoalveolar lavage (BAL) is a diagnostic method for the assessment of the lower respiratory airway health status in horses. Differential cell count and sometimes also total nucleated cell count (TNCC) are routinely measured by time-consuming manual methods, while faster automated methods exist. The aims of this study were to compare: 1) the Sysmex XN-V body fluid (BF) mode with the manual techniques for TNCC and two-part differential into mononuclear and polymorphonuclear cells; 2) the Olympus VS200 slide scanner and software generated deep-learning-based algorithm with manual techniques for four-part differential cell count into alveolar macrophages, lymphocytes, neutrophils, and mast cells. The methods were compared in 69 clinical BAL samples. Results: Incorrect gating by the Sysmex BF mode was observed on many scattergrams, therefore all samples were reanalyzed with manually set gates. For the TNCC, a proportional and systematic bias with a correlation of r = 0.79 was seen when comparing the Sysmex BF mode with manual methods. For the two-part differential count, a mild constant and proportional bias and a very small mean difference with moderate limits of agreement with a correlation of r = 0.84 and 0.83 were seen when comparing the Sysmex BF mode with manual methods. The Sysmex BF mode classified significantly more samples as abnormal based on the TNCC and the two-part differential compared to the manual method. When comparing the Olympus VS200 deep-learning-based algorithm with manual methods for the four-part differential cell count, a very small bias in the regression analysis and a very small mean difference in the difference plot, as well as a correlation of r = 0.85 to 0.92 were observed for all four cell categories. The Olympus VS200 deep-learning-based algorithm also showed better precision than manual methods for the four-part differential cell count, especially with an increasing number of analyzed cells. Conclusions: The Sysmex XN-V BF mode can be used for TNCC and two-part differential count measurements after reanalyzing the samples with manually set gates. The Olympus VS200 deep-learning-based algorithm correlates well with the manual methods, while showing better precision and can be used for a four-part differential cell count

Photoelectron spectromicroscopy at chalcopyrite films

CuInSe2 films were prepared by MBE on GaAs 111 A substrates. ZnSe and ZnO are subsequently deposited in situ by MOMBE. Interface parameters like band offsets and morphology are studied by X ray photoelectron spectroscopy XPS and Low energy electron diffraction LEED . Spectroscopic XPEEM X ray Photo electron emission microscopy at the U49 2 PGM2 beamline at BESSY was used to investigate the lateral homogenity of the interface. After annealing in situ a lateral inhomogenious In diffusion is observed into the ZnSe ZnO interfac

Evaluation of the LaserCyte: an in-house hematology analyzer for dogs and cats

In the present study, the LaserCyte instrument, a fully automated flow cytometer for use in veterinary practice, was evaluated for dogs and cats. Precision (coefficient of variation, CV) for red blood cell (RBC) parameters was ≤3.9%, for reticulocytes between 14.9 and 102%, for white blood cells (WBC) between 3 and 9.5%, for neutrophils between 3.9 and 6.5%, for lymphocytes between 7 and 17.9%, for monocytes between 4.9 and 13.1%, for eosinophils between 10.4 and 32.1%, for basophils between 7.8 and 32%, for platelets between 3.1 and 13.2%, and for platelet indices between 0 and 28.2%. The range of linearity extended the reference ranges. The agreement with reference methods (coefficient of correlation, r) were ≥0.96 (RBC), ≥0.94 (hematocrit), ≥0.96 (hemoglobin), ≥0.95 (mean corpuscular volume), ≥0.94 (WBC), ≥0.93 (neutrophils), ≥0.77 (lymphocytes), ≥0.77 (monocytes), ≥0.29 (eosinophils), ≥0.03 (basophils), ≥0.13 (reticulocytes), and ≥0.86 (platelets). The LaserCyte allowed the correct assessment of RBC and WBC parameters with respect to clinical relevance in the majority of samples. Lymphocytopenia was detected in only 51 out of 89 cases and monocytopenia in one out of 11 cases. The reticulocyte counts were correctly estimated in 85 out of 149 cases. It was concluded that the LaserCyte allowed reliable determination of the RBC parameters, WBCs, neutrophils in both species and platelets in dogs. Based on its capability to reliably determine feline platelets and of the parameters mentioned above, this instrument is considered a useful in-house analyzer for the veterinary practice. Qualitative microscopic assessment of blood smears is still necessary for detecting abnormal cell morphologies, certain cell precursors and blood parasite

Prospective Investigation of Feline Leukemia Virus Infection in Stray Cats Subjected to a Trap–Neuter–Return Program in Switzerland

Feline leukemia virus (FeLV) remains a serious concern in some countries despite advances in diagnostics and vaccines. FeLV-infected cats often have reduced lifespans due to FeLV-associated diseases. The infection is transmitted through social interactions. While Northern European countries have reported a decrease in FeLV among pet cats, Switzerland’s rates remain stagnant at 2.7% (2016/17: 95% CI 1.4–5.2%). Research on FeLV in Swiss stray cats has been lacking, even though these animals could serve as a virus reservoir. Sampling stray cats that do not receive regular veterinary care can be challenging. Collaboration with the Swiss Network for Animal Protection (NetAP) allowed for the prospective collection of saliva samples from 1711 stray cats during a trap–neuter–return program from 2019 to 2023. These samples were tested for FeLV RNA using RT-qPCR as a measure for antigenemia. Viral RNA was detected in 4.0% (95% CI 3.1–5.0%) of the samples, with 7.7% (95% CI 4.9–11.3%) in sick cats and 3.3% (95% CI 2.4–4.4%) in healthy ones. We identified three geographically independent hotspots with alarmingly high FeLV infection rates in stray cats (up to 70%). Overall, including the previous data of privately owned cats, FeLV-positive cats were scattered throughout Switzerland in 24/26 cantons. Our findings underscore welfare concerns for FeLV infections among stray cats lacking veterinary attention, highlighting the potential risk of infection to other free-roaming cats, including those privately owned. This emphasizes the critical significance of vaccinating all cats with outdoor access against FeLV and developing programs to protect cats from FeLV infections

Feline leukaemia virus: half a century since its discovery

In the early 1960s, Professor William (Bill) F.H. Jarrett was presented with a timeGÇôspace cluster of cats with lymphoma identified by a local veterinary practitioner, Harry Pfaff, and carried out experiments to find if the condition might be caused by a virus, similar to lymphomas noted previously in poultry and mice. In 1964, the transmission of lymphoma in cats and the presence of virus-like particles that resembled GÇÿthe virus of murine leukaemiasGÇÖ in the induced tumours were reported in Nature. These seminal studies initiated research on feline leukaemia virus (FeLV) and launched the field of feline retrovirology. This review article considers the way in which some of the key early observations made by Bill Jarrett and his coworkers have developed in subsequent years and discusses progress that has been made in the field since FeLV was first discovered

First evidence of Candidatus Neoehrlichia mikurensis in Hungary

Altogether 2004 Ixodes ricinus ticks, from 37 places in Hungary, were analysed in pools with a recently developed multiplex real-time PCR for the presence of Candidatus Neoehrlichia mikurensis and for other representatives of the genus. Ca. Neoehrlichia mikurensis was identified in nine sampling sites, indicating three separated endemic regions along the borders of Hungary. In addition, results of samples from seven places (except for the western part of the country) were positive in the genus-specific (Ca. Neoehrlichia sp.) PCR, but were negative for Ca. Neoehrlichia mikurensis

Validation of the Sysmex XN-V Automated Nucleated Red Blood Cell Enumeration for Canine and Feline EDTA-Anticoagulated Blood

The enumeration of nRBCs (nucleated red blood cells) by manual counting is time-consuming and imprecise. As the first veterinary hematology analyzer, Sysmex XN-V provides automated nRBC counts. This study aimed to evaluate the performance of Sysmex XN-V in the enumeration of nRBCs for cats and dogs by comparing automated nRBC counts to manual counts from a total of 3810 canine and 2844 feline specimens. Repeatability, reproducibility, stability, carry-over, and linearity were assessed. The repeatability and reproducibility of Sysmex XN-V were good, with mean coefficients of variation (CV) of 4.5% and 5.4%, respectively. Bland–Altman difference analysis revealed mean biases shown as nRBCs/100 WBCs of 0.01 in dogs and 0.11 in cats with low nRBCs (20 nRBCs/100 WBCs). The total observable error was below 9% in both species and at all ranges. Overall concordance between methods was high (91% in canine and 93% in feline samples). The automated nRBC count by Sysmex XN-V was found to be accurate and precise and can replace manual counts for cat and dog samples. Non-statistical quality assurance by scattergram evaluation, re-gating, and confirmation by blood smear evaluation is, however, recommended, especially in cases with severe normoblastosis. This advancement will save time, reduce errors, and add prognostic value to hematological results for animal patients

Acidification is required for calcium and magnesium concentration measurements in equine urine

Background: Acidification of equine urine to promote dissociation of ion complexes is a common practice for urine ion concentration measurements. The objective of this study was to evaluate the effect of acidification and storage after acidification on calcium (Ca), magnesium (Mg) and phosphate (P) concentrations and on fractional excretion (FE) of these electrolytes. Thirty-two fresh equine urine samples were analysed between December 2016 and July 2020. Complete urinalysis (stick and sediment) was performed on all samples. Ca, Mg, P and creatinine concentrations were measured in supernatant of centrifuged native urine, urine directly centrifuged after acidification and urine centrifuged 1 hour after acidification. Urine was acidified with hydrochloric acid to reach a pH of 1–2. Ca, Mg, P and creatinine concentrations were also measured in blood plasma, and fractional excretion of each electrolyte was calculated. Equality of medians was tested with Friedman tests and Bland-Altman bias plots were used to show the agreement between conditions. Results: Acidification had a statistically significant effect on Ca and Mg concentrations, FE$_{Ca}$ and FE$_{Mg}$. Bland-Altman plot revealed a strong positive proportional bias between Ca concentration in native and acidified urine with a mean bias of 17.6 mmol/l. For Mg concentration, the difference between native and acidified urine was small with a mean bias of 1.8 mmol/l. The increase in FE$_{Ca}$ was clinically relevant. Storage of acidified urine had no effect on any of the measured ion concentrations. All P concentrations in native urine samples were below the detection limit of the assay and statistical analysis and calculation of FE$_{P}$ was not possible. Conclusions: Urine acidification is essential for accurate measurement of Ca and Mg concentrations and therefore FE calculations in equine urine. Storage time of 1 hour after acidification does not significantly change Ca and Mg concentrations