Clinical and Clinico-Pathological Observations of the Erythrocyte Sedimentation Rate in Dogs Affected by Leishmaniosis and Other Inflammatory Diseases.

The erythrocyte sedimentation rate (ESR) has been used in canine medicine in several disorders, above all, to evaluate levels of inflammation. This study evaluated the ESR in canine leishmaniosis (CanL) and other inflammatory conditions. Three groups of dogs were examined: CanL affected dogs without clinical signs (INFECTED group, #25) or with clinical signs (SICK group, #43) and dogs affected by acute or acute-on-chronic conditions (OTHER DISEASE group, #65). The ESR was compared with acute phase proteins or reactants either positive or negative (leukogram, fibrinogen, iron, unsaturated iron binding capacity, ferritin, haptoglobin, and albumin) and immunological markers (gamma-globulins, IgG, and IgM). The ESR was higher in the SICK group than in the INFECTED group (median 39 vs. 11 mm/h; p < 0.0001), as well as in the OTHER DISEASE than in the INFECTED groups (median 41 vs. 11 mm/h; p < 0.0001). The ESR appeared outside the reference range for all dogs in the SICK and OTHER DISEASE groups and almost with similar values (mm/h; median 39, 95% CI 31-51 vs. 41, 95% CI 12-87; p > 0.05). The extent of changes in ESR can help to establish the severity of CanL and other inflammatory disorders. As a point-of-care test, the ESR can be used to screen dogs for unhealthy conditions, and its values correlate with the severity of any disease, including CanL.

Analytical Validation of MINI-PET as Point-of-Care for Erythrocyte Sedimentation Rate Measure in Horses.

The erythrocyte sedimentation rate (ESR) is a widely used diagnostic assay in human medicine but nowadays poorly applied in veterinary medicine. This test measures the speed (millimeters per hour) at which red blood cells settle in a whole anticoagulated blood tube. In human medicine, high ESR values are associated with various disorders, including infections, rheumatoid arthritis, oncologic diseases, and other inflammatory conditions. The ESR can also be influenced by some factors such as age and gender. In veterinary medicine, the ESR with the Westergren manual method was almost forgotten over the years due to blood consumption and long turn-around time. The instrument MINI-PET, using a modified Westergren method, does not require blood consumption or release waste product and recently has been applied in canine medicine. The aims of the study in the horse were as follows: to establish the appropriate time to read the ESR with the Westergren reference method; to compare the MINI-PET ESR results with the reference technique; to assess the ESR reference intervals with MINI-PET; and to establish the ESR stability from collection at different time points by MINI-PET. Using 150 horses, we established 60 minutes as the appropriate time for ESR reading with the Westergren method. Moreover, ESR results obtained in 8 minutes with MINI-PET showed a good correlation with the Westergren ESR. Reference intervals (RIs) with MINI-PET were established in mm/h for the healthy horses (geldings 18.6-100.1; stallions, 13.8-55.7; and mares 1-73.7) according to the American Society of Veterinary Clinical Pathology guidelines. In addition, the ESR stability from the blood collection time was evaluated in the MINI-PET on 15 horses: at room temperature, ESR is stable up to 8 hours and at 4°C up to 24 hours. In conclusion, MINI-PET represents a rapid and reliable tool for measuring ESR in horses, offering a valid option to replace the traditional manual technique.

Utility and prognostic significance of leukocyte ratios in dogs with Primary Immune-Mediated Hemolytic Anemia.

Canine immune-mediated hemolytic anemia (IMHA) is a life-threatening condition that is commonly associated with neutrophilia and monocytosis. Leukocyte ratios have been found to have prognostic value in humans and animals affected by a range of inflammatory, infectious, and neoplastic disorders. We hypothesized that in primary IMHA, neutrophil to lymphocyte (NLR), neutrophil to monocyte (NMR), band neutrophil to segmented neutrophil (BNR) and monocyte to lymphocyte (MLR) ratios would be higher in dogs that did not survive to discharge. Medical records of dogs diagnosed with IMHA at two veterinary teaching hospitals were retrospectively reviewed. Twenty-three of the 72 included dogs did not survive to discharge. NLR, NMR, BNR and MLR ratios were compared between dogs that survived to discharge and dogs that died or were euthanized. None of the ratios were significantly different between survivors and non-survivors (P = 0.14-0.99). Area under the Receiver Operating Characteristic (ROC) curve for prediction of non-survival ranged from 0.5 (95% confidence interval 0.38-0.62) for MLR to 0.61 (0.49-0.72) for NMR and was not significantly different from 0.5 for any ratio (P = 0.29-0.99). After exclusion of 31 dogs that received one or both immunosuppressive medications and blood transfusion before presentation, the area under the ROC curve for prediction of survival was significantly different from 0.5 for MLR (0.78, P = 0.01) and NMR (0.78, P = 0.0002). This study suggests that lower MLR and higher NMR may predict poorer prognosis in untreated dogs with IMHA.

Relationship between Blood Parameters and Outcome in Rescued Roe Deer.

Veterinary facility admissions of wild animals are increasing alongside the interest in wildlife diseases. To improve animal welfare, it is therefore important to increase veterinarians' knowledge of wild animal medicine and to improve the clinical and diagnostic procedures, especially in the case of patients affected by trauma or multiple traumas. Blood analysis can be a quick and minimally invasive way of gathering useful clinical information for adequate treatment and management, and, together with a good clinical examination, to help predict hospitalisation outcomes. Few papers have reported reference ranges for the haematological and biochemical parameters of roe deer. This study evaluates the haematological and biochemical parameters in traumatised roe deer in relation to animal hospitalisation outcomes. The study was carried out on a cohort of 98 roe deer divided into groups according to their age and hospitalisation outcome. For each animal, a panel of haematological and biochemical parameters was performed. Significant differences were found between unweaned (<4 months old) groups in terms of MCV, MCH, CK, creatinine, total bilirubin, direct bilirubin, and indirect bilirubin, and between weaned (>4 months old) groups for total bilirubin. Creatine kinase, creatinine, and bilirubin may be useful indicators to correlate with the severity of trauma and help predict prognosis.

Detection of Anti-Erythrocyte Antibodies in Dogs with Inflammatory Bowel Disease (IBD).

Several extra-intestinal manifestations, including immune-mediated cytopenias, are reported in human inflammatory bowel disease (IBD), whereas they are poorly documented in dogs. Hypothesizing that immune-mediated subclinical anemia can occur in canine IBD, the study aim was to evaluate the erythrogram and the presence of anti-RBC antibodies in dogs with IBD. IBD was diagnosed according to the following criteria: chronic gastrointestinal signs, ruling out of extra-intestinal diseases, no improvement with diet trial, histological evidence of inflammatory infiltration, and improvement after immunosuppressant therapy. Canine Chronic Enteropathy Clinical Activity Index (CCECAI) endoscopic and histopathological scores were assessed for each dog. Twenty-five dogs were enrolled, and each dog had a CBC evaluation prior to endoscopy. The CBC was performed using laser hematology analyzer and blood smears were carefully reviewed for the presence of nucleated RBC, anisocytosis, polychromasia, and Howell-Jolly bodies. IgG and IgM anti-RBC antibodies were evaluated with flow cytometry. A high frequency of positive cases for anti-RBC antibodies in dogs with IBD (17/25 dogs) was ascertained. Approximatively 50% of dogs showed some hematologic features of RBC regeneration in addition to hematologic findings consistent with chronic inflammation. Anti-RBC antibodies and signs of erythroid regeneration may suggest possible subclinical chronic immune-mediated hemolysis that can cause anemia in dogs with IBD, together with the chronic inflammation.

Multiple myeloma and primary erythrocytosis in a dog.

A 13-year-old spayed female mixed breed dog was referred for impaired ambulation, limb tremors, back pain, hypergammaglobulinemia on cellulose acetate electrophoresis, and mild proteinuria. Conventional radiology and magnetic resonance imaging (MRI) suggested multifocal neoplastic bone lesions. At the referral examination, lameness and bright red mucous membranes were observed. Severe erythrocytosis, a monoclonal peak in the ß-2 globulin detected by capillary zone electrophoresis, severe proteinuria, bone marrow infiltration of plasma cells, and low serum erythropoietin concentrations were reported. The final diagnosis was multiple myeloma associated with severe primary erythrocytosis. This presentation in a dog is interesting because the combination of both disorders is rare in humans and has not been reported in dogs. Key clinical message: Although rare, multiple myeloma and primary erythrocytosis can occur together in dogs.

Myélome multiple et érythrocytose primaire chez un chien. Une chienne de race mixte stérilisée âgée de 13 ans a été référée pour troubles de la marche, tremblements des membres, maux de dos, hypergammaglobulinémie à l'électrophorèse sur acétate de cellulose et protéinurie légère. La radiologie conventionnelle et l'imagerie par résonance magnétique (IRM) suggéraient des lésions osseuses néoplasiques multifocales. Lors de l'examen de référence, une boiterie et des muqueuses rouge vif ont été observées. Une érythrocytose sévère, un pic monoclonal de la globuline ß-2 détecté par électrophorèse capillaire, une protéinurie sévère, une infiltration de la moëlle osseuse par des plasmocytes et de faibles concentrations sériques d'érythropoïétine ont été rapportés. Le diagnostic final était un myélome multiple associé à une érythrocytose primaire sévère. Cette présentation chez un chien est intéressante car l'association des deux conditions est rare chez l'homme et n'a pas été rapportée chez le chien.Message clinique clé :Bien que rares, le myélome multiple et l'érythrocytose primaire peuvent survenir simultanément chez le chien.(Traduit par Dr Serge Messier).

Leukocytes Ratios in Feline Systemic Inflammatory Response Syndrome and Sepsis: A Retrospective Analysis of 209 Cases.

Sepsis is a challenging condition in which hematological prognostic and diagnostic markers in cats are limited. The aims of this study were to test if there are any differences in leukocyte ratios (NLR, BLR and BNLR) between healthy, SIRS and septic cats (sick cats), and if, within sick cats, NLR, BLR and BNLR may be prognostic markers. A retrospective medical database study included 76 healthy cats (blood-donors), 54 SIRS and 79 septic cats. SIRS group was defined if cats fulfilled SIRS criteria. Sepsis was confirmed with an infectious focus on cytology or a positive culture for bacterial infection. Leukocyte ratios were compared among the three study groups and between survivors and non-survivors in sick cats. NLR resulted significantly higher in the sick group compared to healthy cats (p < 0.0001), although NLR was not different between SIRS and sepsis. An NLR > 4.53 had a sensitivity of 76% and a specificity of 93.4% to detect SIRS/sepsis (OR 44.8 95%CI 17-107). Only BLR and BNLR were significantly different between SIRS and sepsis. NLR was associated with mortality in the sick group (p = 0.04). Although NLR resulted higher in sick cats than healthy, BLR and BNLR demonstrated as promising tools in differentiating SIRS from sepsis. NLR was associated with mortality in sick cats.

Erythrogram Patterns in Dogs with Chronic Kidney Disease.

Anemia is considered a common finding in dogs with chronic kidney disease (CKD), typically as normochromic, normocytic, and non-regenerative. Although anemia can occur at any CKD IRIS (International Renal Interest Society) stage, its severity is related with the loss of kidney function. The aim of the present study was to retrospectively evaluate quantitative and morphological abnormalities of the erythrogram in dogs at different CKD IRIS stages. A total of 482 CBCs from 3648 initially screened were included in the study. Anemia was present in 302/482 (63%) dogs, in the majority of which it was normochromic, normocytic, and non-regenerative (295/302; 98%). The number of reticulocytes was <60,000/µL in the majority of dogs (248/295; 84%), with a correlation between poor regeneration rate and progression of CKD (p = 0.0001). The frequency of anemia significantly differed (p = 0.0001) among the IRIS stages: 108/231 (47%) in IRIS 2, 77/109 (71%) in IRIS 3, and 117/142 (82%) in IRIS 4. Dogs at IRIS stages 3 and 4 were more likely to have moderate to severe anemia, compared to dogs at IRIS stage 2 (p = 0.0001). Anisocytosis was the most frequent morphological abnormality (291/482; 60%), whereas the presence of poikilocytosis showed an association with progression of IRIS stages (p = 0.009). Among different morphological abnormalities, the frequency of fragmented red blood cells and Howell-Jolly bodies showed a significant association with the progression of CKD. Anemia was a frequent finding in CKD dogs, mostly associated with none to poor regeneration rate. Similar to human medicine, advanced CKD stages are more frequently characterized by morphological alterations, such as fragmented red blood cells and Howell-Jolly bodies, which may suggest a more severe condition of reduced bone marrow activity and microangiopathy.

A Retrospective Multicentric Study of Electrochemotherapy in the Treatment of Feline Nasal Planum Squamous Cell Carcinoma.

Feline squamous cell carcinoma (SCC) is currently treated with surgery, radiation therapy and electrochemotherapy (ECT). Both the efficacy and/or safety of ECT were evaluated as a sole therapy with bleomycin to treat feline nasal planum SCC (npSCC). Sixty-one cats were enrolled. Local treatment response was evaluated as complete remission (CR), partial remission (PR) or stable disease (SD). Recurrence rate (RR), disease-free interval (DFI) and progression free survival (PFS) were calculated. A six-point scale was used for ECT toxicity. The median tumor size was 1.5 cm. CR was achieved in 65.6% of cases, PR in 31.1% and SD in 3.3%. The overall response rate was 96.7%, RR was 22.5%, median DFI was 136 days, and median PFS was 65.5 days. ECT toxicity was ≤2 in 51% of cats. Tumor recurrence/progression (p = 0.014) and local treatment response (PR: p < 0.001; SD: p < 0.001) influenced survival time. Cats with toxicity >2 showed a higher probability of tumor recurrence/progression. Tumor-related death was higher in cats with PR (p < 0.001) and recurrence/progression (p = 0.002), in ECT treatment with 1 Hz (p = 0.035) and 1200 V/cm (p = 0.011) or 1300 V/cm (p = 0.016). Tumor size influenced local treatment response (p = 0.008) and toxicity (p < 0.001). ECT is an effective treatment for feline npSCCs and should be considered as the first-line procedure for low-stage tumors.

Platelet abnormalities and platelet-to-lymphocyte ratios in canine immunosuppressant-responsive and non-responsive enteropathy: A retrospective study in 41 dogs.

Few studies have examined platelet alterations in dogs with chronic enteropathy. Our aim was to investigate platelet count (PLT), mean platelet volume (MPV), and platelet-to-lymphocyte ratio (PLR) in dogs diagnosed with immunosuppressant-responsive enteropathy (IRE). In this retrospective study of 41 dogs, data regarding signalment, canine chronic enteropathy clinical activity index (CCECAI), endoscopic and histopathological scores, PLT, MPV, PLR, total serum protein concentrations, albumin, and iron were collected. Clinical response and relapse were assessed with the evaluation of CCECAI over time. One month after starting therapy, dogs with >25% CCECAI reduction were considered responders. During a three-month CCECAI evaluation as part of a twelve-month follow-up, a CCECAI >3 together with a ≥2 unit increase in responder dogs was considered a relapse. PLT and PLR displayed significant negative correlation with MPV. MPV was positively correlated with total protein and albumin levels and negatively correlated with CCECAI. Three dogs were classified as non-responders, and 14 relapsed within 12 months. No differences were observed in PLT, MPV, or PLR between responding/non-responding and relapsing/non-relapsing groups. PLT, MPV, and PLR correlated with total protein, albumin, and CCECAI, confirming PLT as a potential marker, and suggesting MPV as a new marker of clinical efficacy against canine IRE.

Incidental Detection of Onchocerca Microfilariae in Donkeys (Equus asinus) in Italy: Report of Four Cases.

This paper reports the occurrence for the first time in Italy of autochthonous Onchocerca infection in donkeys. Four jennies, bred on the same farm, were referred to the Veterinary Teaching Hospital of Pisa for a check-up on ovarian activity (n = 3) or for veterinary support during the delivery (n = 1). Microfilariae were incidentally detected during the blood smear examination of one jenny. Peripheral blood samples were then collected from the other three jennies and the presence of microfilariae was investigated by Knott's test. Circulating unsheathed microfilariae were identified in all the animals. The level of microfilaraemia was between 1 and 31 microfilariae in 2 mL of blood. Hematological changes showed moderate eosinophilia in one case or both remarkable eosinophilia and basophilia in another case. Based on molecular findings by PCR and sequencing, the microfilariae showed 98% sequence similarity with Onchocerca sp. in the NCBI GenBank database (Accession No.: MK541848.1). The present report provides evidence that Onchocerca is an etiological agent of parasitic infection in donkeys in Italy. Our findings highlight the importance of screening in donkeys for Onchocerca even in the absence of clinical indications.

Are Leukocyte and Platelet Abnormalities and Complete Blood Count Ratios Potential Prognostic Markers in Canine Sepsis?

Background: Sepsis is a common disease in which early diagnosis and prognosis assessment are the main aims in order to arrange a prompt and effective treatment. Objectives: (1) To compare leukogram parameters (WBC, segmented and band neutrophils, lymphocytes, monocytes), platelet count (PLT), mean platelet volume (MPV), and some leukocyte/platelet ratio such as NLR, NBNLR, PLR, and MLR between dogs with systemic inflammatory response syndrome (SIRS) and sepsis. (2) To investigate any difference in the trend of these latter parameters between survivors and non-survivors septic dogs. Animals: 57 dogs with confirmed sepsis and 57 dogs with non-septic SIRS. Methods: A review of the medical records was conducted in order to find dogs with sepsis. Sepsis was defined as the presence of an infectious focus with fulfillment of systemic inflammatory response syndrome criteria (SIRS). Septic dogs had to have a CBC at admission and another CBC within 48 h from the previous timepoint. Purebreds with CBC breed-related abnormalities were excluded, together with dogs without confirmed sepsis and dogs with only a single CBC. NLR, NBNLR, PLR, and MLR were calculated. Univariate analysis of all blood parameters studied was assessed between SIRS and septic dogs. Generalized Estimating Equations models for repeated measures were used to test if the blood parameters studied were modified between survivors and non-survivors in the septic group. Results: Septic dogs had lower median segmented neutrophils count and NLR compared to SIRS dogs (p = 0.02 and p = 0.04, respectively). Lastly, septic dogs had a higher prevalence of toxic neutrophil than SIRS dogs (p = 0.01). We found that for a 1-unit increase of PLR and MLR, the risk of death increased by 50.5 and 60%, respectively. Conclusion and Clinical Importance: Evaluation of NLR at hospital admission may be a useful marker of inflammation, although it showed low sensitivity in differentiating SIRS and septic dogs. The monitoring of some CBC parameters, especially PLR and MLR may be useful in the establishment of prognosis in septic dogs.

Neutrophil-to-Lymphocyte Ratio (NLR) in Canine Inflammatory Bowel Disease (IBD).

Inflammatory bowel disease (IBD) is a multifactorial chronic inflammatory disorder leading to structural changes in the intestinal wall. In humans, the neutrophil-to-lymphocyte ratio (NLR) has been proposed as a promising marker of IBD. This study evaluated the possible clinical and prognostic significance of the NLR in dogs with IBD. This retrospective study enrolled 41 dogs diagnosed with IBD presented to University of Pisa from January 2017 to January 2018. For each dog, age, sex, canine chronic enteropathy clinical activity index (CCECAI), endoscopic and histopathological grading were recorded. Complete blood count, serum total protein, albumin, cholesterol, and C-reactive protein at the time of endoscopy were recorded. A control group (CG) of healthy dogs from a blood donor database was built. NLR was calculated for both IBD and CG as the ratio between absolute neutrophils and lymphocytes. Presence of crypt distension, lacteal dilation (LD), mucosal fibrosis, intraepithelial lymphocytes was recorded. Follow-up information was obtained from electronic medical records and dogs were classified as responders and non-responders based on CCECAI variation between admission and the first recheck. IRE dogs showed higher NLR compared to healthy dogs. NLR correlated negatively with total protein, albumin, and cholesterol and correlated positively with CCECAI. Dogs with LD showed higher NLR than dogs without LD. Non-responders showed higher NLR compared to responders. In conclusion, as in IBD human patients, the NLR acts as an inflammatory marker providing further information on severity of the disease and could be useful in predicting treatment response.

Detection of Leishmania infantum DNA by real-time PCR in saliva of dogs.

This study developed a real-time quantitative PCR (qPCR) assay to detect L. infantum kinetoplast DNA (kDNA) in canine saliva. The qPCR showed an efficiency of 93.8%, a coefficient of correlation of 0.996 and a detection limit of 0.5 fg/reaction (0.005 parasites), although it detected until 0.25 fg/reaction (0.0025 parasites). When samples from 12 dogs experimentally infected with L. infantum were collected, L. infantum kDNA was detected at 16-weeks post-infection (wpi) in 41.7% and 91.7% of saliva and bone marrow samples, respectively, and at 47-wpi in 75% of both samples. L. infantum kDNA can be detected by qPCR in canine saliva, with lower sensitivity in the early stages of infection and a lower parasite load estimation compared to bone marrow. However, saliva had similar sensitivities to bone marrow in the later stages of the infection and could be used to detect L. infantum kDNA being aware of its limitations.

Preventive measures of canine leishmaniosis in Italy: Attitudes of veterinarians based on a questionnaire.

There are several topical ectoparasiticides, drugs, supplements and vaccines that protect dogs against sand fly bites and/or reduce the risk of Leishmania infantum infection. This large variety of products and the absence of comparative studies mean that veterinarians are often faced with uncertainty. The aim of the study was to characterise the attitudes of veterinarians towards leishmaniosis prevention in client-owned dogs in Italy. A web-based questionnaire was prepared with 28 questions covering the working environment of veterinarians, advice given to owners regarding topical ectoparasiticides, advice on systemic prevention, and the use of diagnostic tests on which to base advice for prevention. The questionnaire was emailed to 9,426 veterinarians and answers were collected after 100 days. A total of 542 questionnaires were returned (response percentage 5.8 %): 54.8 % of the veterinarians considered their working area to be endemic, 29.3 % examined dogs from urban areas, and 42.3 % diagnosed ≥10 dogs/year with leishmaniosis. Those veterinarians who diagnosed ≥10 dogs/year with leishmaniosis were more likely to consider their working area to be endemic, whereas those who examined dogs mainly from urban areas were less likely to consider the area to be endemic. Veterinarians who considered the working area to be endemic were more prone to prescribe ectoparasiticides throughout the year, including collars and spot-on products and a combination of the two, vaccine alone or vaccine and domperidone combined, and used qualitative serological Leishmania tests. The attitude of veterinarians did not differ whether dogs were from urban or rural areas. In conclusion, veterinarians who consider their working area to be endemic or who frequently diagnose dogs with leishmaniosis tend to adopt more comprehensive strategies, and their attitudes are not influenced by the origin of the dogs: urban or rural. Overall, in Italy the attitudes of veterinarians in relation to canine leishmaniosis prevention appear to vary considerably.

Procalcitonin Detection in Veterinary Species: Investigation of Commercial ELISA Kits.

In human medicine, procalcitonin (PCT), the precursor of calcitonin, is used for the rapid identification of the origin and severity of sepsis. In veterinary medicine, PCT has been studied in horses, cattle, and dogs, but the use of PCT in diagnostic and/or prognostic settings is not possible because of the lack of validated assays to obtain reference ranges. The aim of the present study was the investigation of commercially available ELISA kits for the detection of canine and equine PCT in plasma samples. Validation of the ELISA kits was performed by using species-specific recombinant proteins spiked both in plasma and buffer samples; linearity, limit of detection (LOD), recovery, and intra-assay and inter-assay variability were calculated. Moreover, clinical samples obtained from sick and healthy animals were also analyzed with the tested kits. Canine PCT was measured with a recombinant canine and a canine PCT ELISA kit. Equine PCT was measured with an equine and a human ELISA PCT kit. Our data demonstrate that the canine recombinant PCT ELISA kit can be used to measure canine PCT in plasma samples, showing an intra-assay and inter-assay coefficient of variation less than 20% and a LOD of 11 pg/mL, whereas the present results do not support the use of the canine PCT ELISA kit. The human PCT ELISA kit is suitable to detect equine PCT with a LOD of 56 ng/mL, whereas the equine PCT ELISA kit did not detect recombinant equine PCT.

The Canine Erythrocyte Sedimentation Rate (ESR): Evaluation of a Point-of-Care Testing Device (MINIPET DIESSE).

The erythrocyte sedimentation rate (ESR) in canine medicine has been replaced by the evaluation of a few sensitive markers of the acute-phase proteins. The aim of the study was to evaluate the ESR using a point-of-care (MINIPET, DIESSE Diagnostica Senese S.p.A.) device (ESR-MP) and to compare the results with the gold standard Westergren method (ESR-W) in dogs. One hundred and nineteen K3-EDTA blood samples for complete blood count were randomly selected and assayed for ESR. The reference interval (RI) was established using the percentile method. The coefficient of variation (CV) in intra-assay and interassay precision of ESR-MP was calculated. The analytical sensitivity (Se), specificity (Sp), positive predictive values (PPVs), and negative predictive values (NPVs) were calculated. Agreement between ESR-MP and ESR-W was assessed with Pearson correlation coefficient (r), Cohen concordance test (K), Passing-Bablok regression, and Bland-Altman plots. Ten canine samples (8.4%) were ruled out because of flag-error by the MINIPET instrument (4.2%) or because they showed the diphasic pattern in ESR-W (4.2%). The canine RI of ESR-MP was 0-10 mm/h. Precision was excellent in intra-assay (CV = 0.02) and interassay (CV = 0.32). The analytical characteristics of ESR-MP in nonanemic samples were as follows: Se = 0.82, Sp = 0.95, PPV = 0.82, and NPV = 0.95. The accuracy of ESR-MP was better than ESR-W in nonanemic samples (r = 0.87; K = 0.77) and lower in anemic subjects (Hct <37%) (r = 0.76; K = 0.69). The Passing-Bablok regression showed the presence of systematic error and the absence of proportional error only in nonanemic blood samples. The Bland-Altman plots gave negative average values due to the difference in RIs and an agreement in both ESRs. The ESR-MP results can be obtained with the same K3-EDTA tubes used for the blood count, in shortcut time, and at reduced costs using the MINIPET device. These investigations highlight that ESR-MP could be useful in canine clinical settings.

Prevalence of Dal blood type and dog erythrocyte antigens (DEA) 1, 4, and 7 in canine blood donors in Italy and Spain.

BACKGROUND: The aim of this study was to determine the prevalence of Dal, and DEA 1, 4, 7 blood types, in a population of canine blood donors from Italy and Spain. Three hundred and twenty blood donor dogs receiving an annual health evaluation were included in the study. DEA 1 blood type was determined using an immunochromatographic strip technique while Dal, DEA 4 and 7 blood types were determined with polyclonal antisera using agglutination on gel columns. RESULTS: Out of 320 dogs blood typed 7 (2 Cane Corso and 5 Doberman Pinschers) (2.2%) were Dal negative; 137 (42.8%) were positive for DEA 1; 320 (100%) were positive for DEA 4 and 43 (13.4%) were positive for DEA 7. CONCLUSION: This study showed a similar prevalence of DEA 1, 7 and 4 to that reported in previous studies in the same, and in different, geographic areas, and provides new data on the prevalence of the Dal blood group in Italy and Spain. There was no significant difference (P = 0.8409) between prevalence of Dal negative blood types found in our population (2.2%) and the prevalence reported in a canine blood donor population from the USA (2.5%). Our study identified Dal negative dogs in a previously tested breed i.e. Doberman Pinschers, but also the Cane Corso breed was found to have Dal negative dogs.

The guaiac-based fecal occult blood test in healthy dogs: Evaluation of the effects of diet, and the ability of the test to detect fecal occult blood.

BACKGROUND: The guaiac-based fecal occult blood test (gFOBt) has been used for human colorectal cancer screening. It can detect the fecal occult blood (FOB) in dogs after oral administration of 20 mg of hemoglobin/kg body weight of blood and is influenced by diet. OBJECTIVES: The aims were to evaluate the effect of diet and the ability of gFOBt to detect FOB in healthy dogs after oral administration of autologous blood. METHODS: Five healthy dogs were fed Purina Hypoallergenic (HA) and Gastrointestinal (EN) diets. Feces were tested with gFOBt before starting diets and at each defecation (hereafter referred to as fecal collection event) throughout the study period. Every 4 days, increased doses of autologous blood were administered orally. The whole blood of one dog was progressively diluted with a saline solution, and dilutions were directly tested with the gFOBt, until a negative result was found. RESULTS: Twelve of 185 (6.5%) gFOBt were found to be positive. No associations between diet and gFOBt positivity were found. A significant association was found between the fecal collection event and gFOBt positivity (P < .001) and between doses of blood and gFOBt positivity (P = .048). The lowest dilution ensuring all positive tests was 6.5 µgHgb /mL. CONCLUSIONS: The gFOBt was not influenced by either the HA or EN diets. The gFOBt positivity was associated with the dose of blood and the fecal collection event. However, caution is needed in the interpretation of results due to the lack of an association between gFOBt positivity and increasing doses of blood in the same dog.

Electrochemotherapy in treatment of canine oral non-tonsillar squamous cell carcinoma. A case series report.

Non-tonsillar squamous cell carcinoma (ntSCC) is a common and locally aggressive oral tumour in dogs. The treatments of choice are currently surgery and radiotherapy. Electrochemotherapy (ECT) is a local ablative anti-tumour technique using electric pulses to enhance the intracellular diffusion of cytotoxic drugs. The aim was to retrospectively evaluate the outcome of patients with oral ntSCC treated with ECT. Twelve dogs with ntSCC were retrospectively enrolled. ECT was combined with IV bleomycin (15 000 UI/m2 ) alone in 11 cases and post-surgery in 1. Parameters considered were: tumour site and size, electroporation parameters, response rate (complete remission [CR], partial remission [PR]), median survival time (MST), recurrence rate (RR), median disease-free interval (DFI) and treatment toxicity (6-point scale). Median tumour size was 1.65 cm (range 0.3-8.0 cm) and the response rate was 90.9% (10/11; 8 CR and 2 PR). Two dogs underwent a second ECT. MST for dogs dead with tumour (n = 2) was 110 days and for dogs dead without tumour (n = 3) was 831 days. Among five surviving dogs, one experienced tumour recurrence and four were in CR. Results from two dogs were analysed separately. Overall RR was 27.3%. DFI and MST for dogs with recurrence were 50 and 115 days, respectively. Treatment toxicity was very low. We noticed that all dogs with tumours smaller than 1-2 cm achieved CR without recurrence suggesting a favourable prognosis when using ECT. ECT for canine ntSCC could be considered a valid treatment option especially for smaller tumours, but a larger caseload would be needed to confirm this statement.