A comparison study between the Siemens ADVIA 120 and manual method for the differential white blood cell count in goats.

BACKGROUND: Although widely used, the ADVIA 120 hematology analyzer has not been previously validated for determining the differential leukocyte count in goats. OBJECTIVES: The aim of this study was to compare the differential leukocyte counts provided by the ADVIA 120 (A-diff) and the manual method (M-Diff) in goats. METHODS: EDTA blood samples that were analyzed within 4 h of collection were used in the study. The following exclusion criteria were applied: inappropriately filled tubes or tubes containing clots, erroneous ADVIA peroxidase cytograms, and blood smears of poor quality. The A-Diff was compared with the M-Diff performed by two independent observers on 200 leukocytes. RESULTS: Forty samples were included after previously excluding eight samples. The correlation between the A-Diff and M-Diff was very strong for eosinophils (r = .870, p < .001) and strong for lymphocytes (r = .796, p < .001) and neutrophils (r = .730, p < .001), while no significant correlation was observed for monocytes (r = .026, p = .872). The Passing-Bablok regression analyses revealed statistically significant constant errors for neutrophils (5.83%; 95% confidence interval [CI]: 0.41%, 12.18%) and eosinophils (1.89%; 95% CI: 1.17%, 2.71%). Bland-Altman analyses showed a statistically significant negative bias for lymphocytes (-5.0%) and a statistically significant positive bias for eosinophils (2.2%). The very low basophil percentages precluded a meaningful method comparison. CONCLUSIONS: The ADVIA 120 overall demonstrated good performance for the differential WBC count in goats under the conditions of this study. Therefore, it can be considered suitable for routine hematologic screening in goats. Nonetheless, it should be emphasized that any abnormal result should be confirmed with a blood smear evaluation.

Systematic map of the most recent evidence (2010-2019) on ruminant production-limiting disease prevalence and associated mortality in Ethiopia.

Background: Ethiopia's livestock sector supports the livelihoods of millions of smallholder farmers. However, despite the improvements of recent years, livestock productivity remains low due to critical constraints, including infectious diseases. The aim of this study was to collate and synthesize the published evidence on ruminant disease frequency and disease-associated mortality in Ethiopia, by identifying knowledge gaps and clusters in the literature to provide the basis for a decision-making tool. Methods: Searches on both bibliographic and organization databases were conducted in English and were restricted to the period 2010-2019. Search results were screened for relevance at title, abstract and full text level. Results: There were 716 articles identified relevant to the research question. The systematic map revealed an increased publication output from 2012-2017, compared to 2010-2011 and 2018-2019. Most studies were conducted in Oromia, Amhara and SNNPR. A substantial body of evidence was found for trypanosomosis, ectoparasite infestation, fasciolosis, nematodiasis, echinococcosis, and brucellosis. Conclusion: This study suggests that despite the high output of epidemiological publications, further understanding of a considerable number of diseases is required and where evidence is abundant, synthesis of information should be carried out in order to better inform decisions on disease control priorities in the livestock sector.

Association of platelet indices with glycemic status in diabetic dogs.

We investigated the platelet count (PLT), mean platelet volume (MPV), and plateletcrit (PCT) in dogs with type 1 diabetes mellitus (DM) compared to healthy controls, and their association with the major fraction of glycated hemoglobin (HbA1c). Blood samples from 33 clinically healthy dogs and 14 newly diagnosed diabetic dogs were included. CBCs were performed with the Advia 120; HbA1c was determined using a validated assay (Capillarys 2 flex-piercing; Sebia). Median [range] PLT and PCT were significantly higher (p = 0.040 and p = 0.010, respectively) in diabetic dogs (434 [176-987] × 109/L and 0.60 [0.26-1.22]%, respectively) compared to healthy dogs (297 [223-671] × 109/L and 0.35 [0.24-0.87]%, respectively]. Thrombocytosis was observed in 6 of 14 (43%) diabetic dogs. The median MPV was not significantly different (p = 0.114) between the diabetic (13.6 fL, 10.1-22.6 fL) and healthy dogs (11.9 fL, 8.6-19.1 fL). A significant, albeit weak, correlation was detected between HbA1c and PLT (rho = 0.298, p = 0.042) and PCT (rho = 0.340, p = 0.019), but no significant correlation was found with MPV (rho = 0.199, p = 0.180). Canine DM was associated with increased PLT and PCT, which was correlated with glycemic status. Our findings suggest dysregulated megakaryopoiesis in diabetic dogs, but this should be confirmed by large-scale studies, and the clinical implications should be investigated.

Prevalence of antinuclear antibodies and rheumatoid factor titers in dogs with arthritis secondary to leishmaniosis (Leishmania infantum).

Dogs with infectious arthritis may occasionally exhibit positive serum antinuclear antibody (ANA) and rheumatoid factor (RF) titers; however, relevant data are sparse for arthritis secondary to canine leishmaniosis (CanL) caused by Leishmania infantum. We determined the prevalence of positive serum ANA and RF titers in dogs with arthritis secondary to CanL. Blood samples from adult, client-owned dogs with purulent arthritis secondary to CanL, without any comorbidities, were collected for diagnostic purposes. Serum ANA titers were measured by immunoperoxidase test and RF titers by the Rose-Waaler latex test. Twelve of 23 dogs enrolled prospectively in our study had clinical arthritis, and 11 of 23 had subclinical arthritis. Based on LeishVet clinical staging, 7 dogs had clinical stage II disease, 11 had clinical stage III disease, and 5 had stage IV. None of the 23 dogs was seropositive for ANA; 3 of 23 were positive for RF. ANA and/or RF seropositivity, in dogs with CanL-associated arthritis, appears to be weak, if present at all. Based on our results, positive serum ANA and RF titers should not be expected in dogs with arthritis secondary to CanL.

The effect of age and sex on glycated hemoglobin in dogs.

We investigated the effect of age and sex on canine glycated hemoglobin (HbA1c) using a validated capillary electrophoresis assay. Aliquots of EDTA blood samples collected for routine health checks were used. HbA1c was measured using the Capillarys 2 flex-piercing system (Sebia). We included 58 clinically and hematologically healthy, normoglycemic dogs (29 males, 29 females), allocated to 3 age groups: young (14 dogs <1-y-old), adult (31 dogs 1-7.9-y-old), and senior (13 dogs ≥8-y-old). The mean (± SD) HbA1c was not significantly different (p = 0.428) between the age groups (young: 1.68 ± 0.54%; adult: 1.59 ± 0.41%; senior: 1.80 ± 0.57%). The HbA1c was not significantly correlated with age (rho = 0.144, p = 0.280). The median (range) HbA1c was not significantly different (p = 0.391) between male [1.7% (0.5-2.5%)] and female [1.5% (1.0-2.7%)] dogs. Age and sex do not appear to affect canine HbA1c; however, a study of geriatric dogs would be needed to fully exclude an effect of age on HbA1c.

Diagnostic and prognostic value of peripheral blood leucocyte ratios in sick cats.

The objective of this study was to assess the diagnostic and prognostic utility of feline neutrophil-to-lymphocyte (NLR) and lymphocyte-to-monocyte ratios (LMR) in a variety of underlying diseases. Five-year medical records from cats presenting to the internal medicine unit of a veterinary teaching hospital were retrospectively reviewed. Cats were considered for inclusion based on complete medical records. ADVIA 120 was used for the complete blood counts; the NLR and LMR were calculated by dividing the absolute numbers of the respective leucocytes. Two hundred and nineteen sick and 20 healthy cats were included in the study. The median NLR and LMR were significantly (P < 0.05) elevated and decreased, respectively, in cats with infectious, neoplastic and chronic kidney diseases compared to controls. Additionally, cats with neoplasia had significantly higher median NLR compared to cats with urinary tract and gastrointestinal diseases. Non-survivors had significantly higher NLR and lower LMR compared to survivors. Both ratios had suboptimal prognostic performance for the outcome of sick cats (NLR sensitivity: 37.9%, specificity: 86.4%; LMR sensitivity: 69.0%, specificity: 61.0%). Many different disease categories were associated with increased NLR and decreased LMR compared to controls, but the overall prognostic performance of the two leucocyte ratios was suboptimal.

Effect of anaemia and erythrocyte indices on canine glycated haemoglobin.

BACKGROUND: The major fraction of glycated haemoglobin (HbA1c) depends on blood glucose concentration and erythrocyte lifespan, and consequently erythrocyte indices may affect HbA1c; our objective was to study this effect in dogs. METHODS: Blood samples from two (healthy and anaemic) age- and sex-matched, normoglycaemic populations were prospectively included. Advia 120 and Capillarys 2 flex-piercing were used for the haematological and HbA1c analyses, respectively. RESULTS: Lower mean HbA1c was found in dogs with regenerative (n = 8, 0.88 ± 0.58%) and non-regenerative anaemia (n = 12, 1.36 ± 0.56%) compared to healthy ones (n = 40, 1.68 ± 0.48%); the difference was significant (p < 0.001) between the healthy dogs and those with regenerative anaemia. HbA1c was significantly correlated (p < 0.05) with red blood cell count (r = 0.506), haemoglobin (r = 0.474), haematocrit (r = 0.467), mean corpuscular volume (r = -0.289), mean corpuscular haemoglobin concentration (r = 0.284), red cell distribution width (r = -0.286) and reticulocytes (rs = -0.542). CONCLUSION: Anaemia, particularly if regenerative, can lower HbA1c. The significant correlation between HbA1c and erythrocyte indices likely reflects the effect of erythrocyte turnover on HbA1c.

A comparison study between the results of the Siemens ADVIA 120 analyzer and the manual method for differential leukocyte counts in sheep.

BACKGROUND: The ADVIA 120 is a widely used hematology analyzer, which has not been previously validated for determining differential leukocyte counts in sheep. OBJECTIVES: We aimed to compare differential leukocyte counts on the ADVIA 120 (A-Diff) with counts obtained using the manual method (M-Diff) in sheep. METHODS: Ethylenediaminetetraacetic acid-anticoagulated blood samples analyzed within 4 hours of collection were used. Samples with inappropriately filled tubes, detectable clots, overtly erroneous ADVIA peroxidase cytograms, and poor-quality blood smears were excluded from the study. Two independent observers compared the results of the A-Diff with those of the M-Diff. The M-diff was performed by counting 200 leukocytes on a blood smear. RESULTS: Overall, 88 samples (44 rams and 44 ewes) were included. The correlation between the A-Diff and M-Diff was high for neutrophils (r = .873, P < .001), lymphocytes (r = .863, P < .001), and eosinophils (r = .750, P < .001), and low for monocytes (r = .212, P = .048). The Passing-Bablok regression analyses revealed constant error for eosinophils [1.17%; 95% confidence interval (CI): 0.67%, 1.55%] and proportional error for lymphocytes (0.84; 95% CI: 0.74, 0.95) and eosinophils (0.85; 95% CI: 0.74, 0.96). The Bland-Altman analyses revealed negative biases of 2.4% and 3.0% for neutrophils and lymphocytes, respectively, and positive biases of 3.2% and 0.8% for monocytes and eosinophils, respectively. The extremely low basophil percentages precluded a meaningful method comparison for this leukocyte type. CONCLUSIONS: The ADVIA 120 appears to perform well for determining neutrophil, lymphocyte, and eosinophil percentages in sheep as compared with the manual method. However, blood smear examinations are recommended for the confirmation of abnormal differential leukocyte count results.

Development of a systematic mapping review protocol for the most recent evidence on ruminant infectious disease frequency and disease-associated mortality: Ethiopia as a case study.

Almost 19% of the GDP of Ethiopia results from livestock production. Ruminants, in particular, form the majority of the national herd and are a critical source of income for smallholder farmers. Infectious diseases have been identified as a major cause of reduced livestock productivity in low- and middle-income countries (LMICs); therefore, a sound and comprehensive understanding of the relevant evidence would be beneficial in order to enable decision making on disease control policies. However, livestock disease data from sub-Saharan Africa is variable and disparate, which poses a challenge for evidence synthesis. This paper describes a protocol for a systematic mapping review of the recent available evidence on ruminant disease prevalence and associated mortality in Ethiopia. Literature sources will be identified using database search strategies. The titles, abstracts and, subsequently, full texts will be screened for inclusion based on predefined eligibility criteria. Specific data will be extracted and a preliminary qualitative assessment of the evidence will be performed using predefined indicators. The planned systematic map will be the first to provide a large-scale overview of the available ruminant disease evidence in Ethiopia; the final output will be an interactive dashboard tool to inform critical stakeholders in policy and research.

Capillary hemoglobin electrophoresis of healthy and anemic dogs: Quantification, validation, and reference intervals of hemoglobin fractions.

Despite the advances in canine medicine and the rapid gaining of attention of canine models in biomedical field and particularly in hemoglobin genes research, the studies on canine hemoglobin composition are sparse with ambiguous findings. Our aim was: i) to investigate the electrophoretic pattern of canine hemoglobin and the possible effects of age, sex, and anemia using a capillary electrophoresis assay, and ii) to validate this assay and calculate reference intervals (RIs) for canine hemoglobin fractions. Blood samples were collected from 53 healthy and 42 dogs with regenerative and non-regenerative anemias. The Sebia Capillarys 2 flex-piercing was used for hemoglobin analysis and it was validated using canine blood samples. R statistical language was employed for the statistical analyses. A major hemoglobin fraction (named HbA0) and a minor one (named HbA2) were identified in 100% and 47.4% of samples, respectively. The within-run and between-run CV was 0.1% for HbA0, and 9.1% and 11.2% for HbA2, respectively. The extremely narrow range of HbA0 and HbA2 values hampered a linearity study using canine blood samples. The RIs for HbA0 and HbA2 were 98.9-100% and 0-1.1%, respectively. HbA0 and HbA2 values were not significantly correlated with age (P = 0.866) or reticulocyte count (P = 0.731). No differences were observed in the median HbA0 and HbA2 between the two sexes (P = 0.887), and healthy and anemic dogs (P = 0.805). In conclusion, the capillary electrophoresis revealed a major hemoglobin fraction and an inconsistently present minor fraction. No effect of age, sex, anemia, or regenerative status of anemia was detected. The assay used was validated and RIs were generated, so as to be suitable for use in future investigations.

Circulating and bone marrow myeloid cells containing Leishmania amastigotes in a case of advanced canine leishmaniosis.

A 5-y-old male Poodle mix was presented with intermittent vomiting, anorexia, and weight loss. Physical examination revealed emaciation, lethargy, dehydration, hypothermia, respiratory distress, and splenomegaly. Based on clinicopathologic, serologic, and parasitologic findings, diagnoses of severe leishmaniosis and dirofilariasis were made. Extracellular, intraneutrophilic, and intramonocytic Leishmania amastigotes were observed on blood smear and buffy coat smear examination. In blood smears, 0.2% of neutrophils were observed to be infected; in buffy coat smears, 0.5% of neutrophils and 0.1% of monocytes were found to be infected. Leishmania amastigotes were also found engulfed by eosinophils and neutrophil precursors in bone marrow aspiration cytology. The detection of Leishmania amastigotes in blood smears is rare, and the clinical significance is uncertain. In circulating blood, Leishmania amastigotes are primarily found phagocytized by neutrophils. Although debatable, there is growing evidence that neutrophils are used as carriers enabling the "silent entry" of the protozoa into macrophages ("Trojan horse" theory). To date, cytologic screening of blood smears for the diagnosis of canine leishmaniosis is not a routine practice. Clinical pathologists and practitioners should be aware that Leishmania amastigotes may be present in neutrophils and less frequently monocytes during blood smear evaluation; neutrophil precursors and eosinophils may also be parasitized in bone marrow specimens.

Hereditary methemoglobinemia in a cyanotic cat presented for ovariohysterectomy.

A 1-year-old, female, domestic shorthair cat with a history of cyanotic mucous membranes for several months was referred for ovariohysterectomy. Blood samples exhibited a noticeably brownish discoloration, while laboratory screening revealed mild-to-moderate erythrocytosis and near normal partial arterial oxygen pressure. Blood methemoglobin content was 41% of total hemoglobin concentration, and erythrocytic methemoglobin reductase activity was < 1% compared with control samples. A diagnosis of hereditary methemoglobinemia was established. After an intravenous injection of methylene blue, the cat's mucous membranes became transiently pink, and the ovariohysterectomy was uneventful. Methylene blue may have improved safety during anesthesia and surgery. Hereditary methemoglobinemia should be considered in persistently cyanotic cats with normal partial arterial oxygen pressure and lack of evidence of cardiopulmonary disease, anemia, or toxin exposure.

Méthémoglobinémie héréditaire chez une chatte cyanotique présentée pour une ovariohystérectomie. Une chatte domestique âgée de 1 an avec une anamnèse de muqueuses cyanotiques pendant plusieurs mois a été recommandée pour l'ovariohystérectomie. Des prélèvements sanguins présentaient une décoloration brune manifeste tandis que les tests de laboratoire ont révélé une érythrocytose de légère à modérée et une pression d'oxygène artérielle partielle presque normale. Le contenu de méthémoglobine sanguine était de 41 % de la concentration totale des hémoglobines et l'activité de la réductase de la méthémoglobine érythrocytaire était < 1 % comparativement aux prélèvements témoins. Un diagnostic de méthémoglobinémie héréditaire a été posé. Après une injection intraveineuse de bleu de méthylène, les muqueuses du chat sont devenues provisoirement roses et l'ovariohystérectomie a été réalisée sans complications. Le bleu de méthylène peut avoir amélioré l'innocuité durant l'anesthésie et la chirurgie.(Traduit par Isabelle Vallières).

The effect of subchronic oral exposure to zearalenone on hematologic and biochemical analytes, and the blood redox status of adult rabbit bucks.

BACKGROUND: Zearalenone (ZEN) is a mycoestrogen with a ubiquitous presence in animal feeds, which also has hematotoxic, hepatotoxic, nephrotoxic, and immunotoxic properties. However, there is a paucity of literature that discusses the effects of ZEN on rabbits. OBJECTIVES: The aim of this study was to evaluate the effect of a prolonged, low-level (50 µg ZEN/kg body weight) exposure on the clinicopathologic and redox status analytes of rabbit bucks. METHODS: Ten adult bucks were included in the study. Each underwent a 7-week control period, followed by a 7-week exposure period. Water or ZEN solutions were daily administered orally (0.5 mL) during the control and exposure periods, respectively. Blood samples were collected weekly for Complete Blood Counts, serum biochemical analyte and reactive oxygen metabolite (ROM) measurements. Data were analyzed using a mixed model, and the level of significance was set at a P of <0.05. RESULTS: During the ZEN exposure period, significant increases were noted in the red blood cell distribution width (RDW) and mean platelet volumes (MPVs), as well as in the white blood cell, monocyte, and eosinophil counts. Significant increases were observed in aspartate aminotransferase and total bilirubin, whereas urea, creatinine, glucose, total calcium, sodium, and potassium concentrations were significantly decreased. The ROM concentrations did not differ significantly between the control and ZEN exposure periods. CONCLUSIONS: Under the present experimental conditions, ZEN affected some of the clinicopathologic analytes of adult rabbit bucks; these changes were mostly indicative of mild hepatocellular damage and dysfunction, inflammatory and/or allergic responses, and renal tubular damage. A ZEN dose of 50 µg/kg body weight did not seem to affect the blood redox status of bucks, as evaluated by the ROM concentrations.

Canine and feline dirofilariosis in a highly enzootic area: first report of feline dirofilariosis in Greece.

Dirofilaria immitis (heartworm) is enzootic in many areas of the world and quite prevalent in southern European countries. Although dogs are the main host of the parasite, cats may also be infected, and the prevalence of feline dirofilariosis is associated with the respective prevalence of canine infection in any given area. The aim of the present study was to investigate the proportion of D. immitis infection among dogs and cats that were not under any kind of prophylactic treatment and were living in a heartworm enzootic area. In total, 180 stray animals (148 dogs and 32 cats) living in a shelter in Northern Greece were examined for heartworm infection by the Knott's test and serology (antigen and in cats also antibody detection), and additionally echocardiography in the infected cats. Thirty-seven (25%, CI 18.7-32.5%) of the dogs and 3 (9.4%, CI 3.2-24.2%) of cats were found to be positive, by at least one of the tests applied. In 2 of the infected cats, the parasites were also detected by echocardiography. One of the positive cats died suddenly 1 year after diagnosis and at necropsy two decomposing D. immitis were found in the right ventricle and pulmonary artery. This is the first report of confirmed feline dirofilariosis in Greece. The detected proportion of infection in cats was 38% of the respective canine infection in the examined shelter. The results of the present study underline the high risk of infection of cats living in enzootic areas and the imperative character of preventive measures in such conditions.

Effect of subchronic oral exposure to zearalenone on the reproductive system of rabbit bucks.

OBJECTIVE To determine the effect of subchronic oral exposure to zearalenone (ZEA) at a daily dose of 50 µg of ZEA/kg of body weight (an environmentally relevant concentration) on the reproductive system of rabbit bucks. ANIMALS 8 healthy sexually mature New Zealand White rabbits. PROCEDURES During the experimental period (March to June), each rabbit underwent a 7-week control protocol and then a 7-week treatment protocol. Water (0.5 mL) or ZEA solution (50 µg/kg [0.5 mL]) was administered orally once daily during the control and treatment period, respectively; ejaculates were collected weekly. Studied end points included semen quality variables (spermatozoa kinetics, morphology, viability, and DNA fragmentation), serum testosterone concentration, and results of histologic examination of the testes and epididymides following euthanasia at the end of the experimental period. RESULTS Treatment with ZEA solution resulted in significant increases in spermatozoa beat-cross frequency, in the percentages of spermatozoa with head and midpiece abnormalities, and in the percentages of DNA-fragmented spermatozoa, compared with effects of the control treatment. Serum testosterone concentration, other spermatozoa velocity variables, and percentages of progressive and total motility, rapidly or slowly moving spermatozoa, and live spermatozoa did not differ significantly between the 2 periods. Histologic examination revealed no patterns of abnormal findings in the testes and epididymides. CONCLUSIONS AND CLINICAL RELEVANCE Oral treatment with ZEA solution at an enviromentally relevant concentration caused minor interference with rabbit bucks' sperm quality. Although mostly considered mild, the sperm quality changes warrant further investigation in terms of fertilizing capacity impairment.