Arianna Miglio

Phylogenetic Relationships of Three Italian Merino-Derived Sheep Breeds Evaluated through a Complete Mitogenome Analysis

In Italy, the crisis of the wool industry triggered the necessity to reconvert the two traditional Merino-derived breeds, Gentile di Puglia and Sopravissana, to meat production, by creating the Merinizzata Italiana. The aim of the present study was to assess the genetic diversity of these three Italian Merino-derived (IMd) breeds by examining the molecular information encoded in the maternally-inherited mitochondrial DNA (mtDNA). A parallel molecular investigation was performed on the putative paternal and maternal breeds, the Merino from Spain and the Appenninica from Italy, respectively, as well as on three unrelated dairy breeds (Sarda and Comisana from Italy, and Lacaune from France). Firstly, the mtDNA control region of 291 samples was analyzed. When comparing the overall genetic distances among the eight stocks, the three IMd breeds clustered together close to the Appenninica, thus confirming its parental role. Among the 90 IMd samples, 82 different haplotypes were observed, almost all belonging to haplogroup B, and only one to A. For 23 mtDNAs, including nine IMd, the analysis was then brought to the level of entire mitogenomes. Three distinct sub-haplogroups within B were found to encompass the IMd samples, with one clade (B1a2a1) apparently restricted to those sheep. Thus, despite experiencing a drastic reduction in number (mainly due to changes in breeding practices driven by the economy), the IMd breeds still represent a reservoir of distinctive mitochondrial variants, which could potentially contribute to the development of conservation and management programs of Italian sheep breeds.

Disseminated angiostrongylosis with massive cardiac and cerebral involvement in a dog from Italy

A case of disseminated angiostrongylosis caused by Angiostrongylus vasorum in a dog living in Italy is here described. The dog was referred for severe respiratory distress and epileptic seizures; clinicopathological findings were consistent with severe pneumonia associated with right-sided heart failure and multifocal involvement of the brain. Bronchoalveolar fluid analysis identified a multitude of nematode larvae, identified as A. vasorum by conventional and biomolecular (PCR) methods. The major anatomo-histopathological lesions were chronic granulomatous pneumonia, a severe multifocal granulomatous myocarditis and multifocal mild vascular and inflammatory disease in the brain. A. vasorum should be included among the differentials of dogs with cardiovascular and neurologic disease.

Signalment and blood types in cats being evaluated as blood donors at two italian university blood banks.

Data from potential feline blood donors presented at two university blood banks in Italy were recorded. Blood typing was performed using an immunochromatographic method. Over the three years of the study 357 cats representing 15 breeds, 45.3% female and 54.7% male, with a mean age of 3.8 years were evaluated. Of these 90.5% were blood type A, 5.6% type B, and 3.9% type AB. The majority of the cats (54.6%) were European DSH (92.3% were type A, 5.1% type B, and 2.6% type AB), and 21% were Maine Coon (MCO) cats (100% blood type A). The estimated frequencies of transfusion reactions following an unmatched transfusion between DSH (donors and recipients), MCO (donor and recipients), DSH donors and MCO recipients, and MCO donors and DSH recipients were 4.8%, 0%, 0%, and 5.1% for major reactions and 7.2%, 0%, 7.7%, and 0% for minor transfusions reactions, respectively. In a population of blood donors that includes DSH and MCO the risk of transfusion reaction is between 5% and 8% if typing is not performed on donor and recipient blood. Blood typing should therefore be performed before transfusion to remove the risk of transfusion reactions due to blood type incompatibilities.

Natural infection of Anaplasma platys in dogs from Umbria region (Central Italy).

Anaplasma platys is a tick-borne pathogen causing the Infectious Canine Cyclic Thrombocytopenia. The pathogenesis of this disease is not yet well understood, due to the wide variety of clinico-pathological patterns described worldwide and to the high prevalence of co-infections with other vector-borne pathogens occurring in endemic areas. The present paper reports 3 cases of infection by A. platys occurring in dogs native to Central Italy, considered a non-endemic area to date. Infections were initially diagnosed based on clinical data and observation of morulae within platelets and then confirmed by biomolecular techniques. Moreover, two dogs showed an immune-mediated hemolytic anemia, as yet not described in literature in association with A. platys infection. The symptoms and the pathological findings observed will be discussed, as well as the importance to include this pathogen in the differential diagnosis of tick-borne diseases even in Central Italy.

Acute undifferentiated leukaemia in a dog

BACKGROUND Acute undifferentiated leukaemia (AUL) is considered a separate entity in the context of acute leukaemias. AUL is extremely rare in both humans and dogs, has a rapid clinical course and does not respond to treatment. It is characterised by the presence of blast cells within the bone marrow and/or peripheral blood at levels ≥ 20% and even up to 100% of all nucleated cells. Blast cells are unable to be differentiated on morphological, cytochemical and phenotypic criteria into myeloid or lymphoid lineages because of their immaturity and/or atypia. CASE REPORT An 8-year-old German Shepherd dog was referred for depression, asthenia, mild anaemia, thrombocytopenia and marked leucocytosis. Abdominal ultrasound showed hepatomegaly, splenomegaly, bilateral nephromegaly and enlargement of mesenteric lymph nodes. Echocardiography revealed biventricular hypertrophy with abnormal tissue density of the myocardium. Blood and bone marrow smears were composed of 95% unclassifiable and/or atypical blast cells and signs of dysplasia of the erythroid and thrombocytic/megakaryocytic lineages were present. Blast cells were negative for all cytochemical stains used and flow cytometry of peripheral blood revealed 85% of total leucocytes consisting of small-to-medium-sized cells, negative for all lymphoid and myeloid markers except CD45 and CD34. After necropsy, cytology and histology revealed that blast cells had diffusely infiltrated all tissues examined. Both erythroid and megakaryocytic extramedullary haemopoiesis was also detected in the spleen, lymph nodes and liver. All immunohistochemical stains used were negative. CONCLUSION On the basis of all the results, a diagnosis of acute leukaemia involving a very primitive haematopoietic precursor was made.

Stored Canine Whole Blood Units: What is the Real Risk of Bacterial Contamination?

Background Bacterial contamination of whole blood (WB) units can result in transfusion‐transmitted infection, but the extent of the risk has not been established and may be underestimated in veterinary medicine. Objectives To detect, quantify, and identify bacterial microorganisms in 49 canine WB units during their shelf life. Animals Forty‐nine healthy adult dogs. Methods Forty‐nine WB units were included in the study. Immediately after collection, 8 sterile samples from the tube segment line of each unit were aseptically collected and tested for bacterial contamination on days 0, 1, 7, 14, 21, 28, 35, and 42 of storage. A qPCR assay was performed on days 0, 21, and 35 to identify and quantify any bacterial DNA. Results On bacterial culture, 47/49 blood units were negative at all time points tested, 1 unit was positive for Enterococcus spp. on days 0 and 1, and 1 was positive for Escherichia coli on day 35. On qPCR assay, 26 of 49 blood units were positive on at least 1 time point and the bacterial loads of the sequences detected (Propionobacterium spp., Corynebacterium spp., Caulobacter spp., Pseudomonas spp., Enterococcus spp., Serratia spp., and Leucobacter spp.) were <80 genome equivalents (GE)/μL. Conclusions and Clinical Importance Most of the organisms detected were common bacteria, not usually implicated in septic transfusion reactions. The very low number of GE detected constitutes an acceptable risk of bacterial contamination, indicating that WB units have a good sanitary shelf life during commercial storage.

Serum protein concentration and protein fractions in clinically healthy Lacaune and Sarda sheep using agarose gel electrophoresis.

BACKGROUND Serum protein electrophoresis (SPE) provides useful information in ruminants, but reference intervals (RI) are different from other species. There have been no reports of SPE RI for dairy sheep using agarose gel electrophoresis (AGE). OBJECTIVE The purpose of the study was to evaluate the serum concentration of total protein (TP) and protein fractions determined by AGE in mid-lactating dairy ewes, to establish RI, and to assess potential differences between Lacaune (L) and Sarda (S) sheep breeds. METHODS Blood samples were collected from healthy, mid-lactating ewes. SPE was assessed using a semi-automated AGE system. Reference intervals (90% confidence intervals) for TP and each protein fraction were determined using the nonparametric method for combined data, and the robust method for data from the single breeds. Data from S and L sheep were compared using the Mann-Whitney U test. RESULTS The 172 sheep included 116 L and 56 S ewes, 2-6 years old. There were significant differences between S and L breeds, and RI were calculated for TP, albumin, α1 -globulin, α2 -globulin, β1 -globulin, β2 -globulin, γ1 -globulin, and γ2 -globulin concentrations, and for the Albumin/Globulin ratio. Group S showed higher concentrations of TP, α2 -, β1 -, β2 -, and γ1 -globulins, whereas L was higher for albumin and γ2 -globulin concentrations, and A/G ratio (P < .05). CONCLUSIONS The resolution with AGE was excellent, allowing standardization of 7 protein fractions, detection of differences between S and L ewes, and determination of RI for French (Lacaune) and Italian (Sarda) dairy sheep.

Haematological and biochemical reference intervals in adult Maine Coon cat blood donors.

Objectives The objectives of this study were to derive Maine Coon haematological and biochemical reference intervals (RIs) from adult healthy blood donors, to validate (or reject) the use of published RIs for the general feline population in this breed, and to evaluate the effects of age, sex and weight on the haematological and biochemical results. Methods Haematological and biochemical data were retrieved retrospectively from a database of 81 healthy adult Maine Coon cat blood donors and were analysed to generate normal RIs. RIs were determined and compared with established non-breed-specific feline RIs according to the Clinical and Laboratory Standards Institute guidelines and the American Society of Veterinary Clinical Pathology guidelines using Reference Value-Advisor (version 2.1) software. Results The age of the cats ranged from 1–8 years (mean 4.4 years), 42 were female and 39 were male, and weights ranged from 4.9–8.5 kg (mean 6.7 kg). New Maine Coon RIs were proposed for red blood cell count, mean corpuscular volume, mean corpuscular haemoglobin concentration, reticulocyte count and percentage. Haematocrit was higher in male cats (mean HCT 42.9% vs 41% in females; P = 0.001) and in heavier cats (P = 0.003; slope 1.0, regression equation HCT = 35.1 + 1.0 × weight). New biochemical RIs were proposed for urea, aspartate aminotransferase, γ-glutamyl transpeptidase (GGT), alkaline phosphatase, total protein and albumin in Maine Coons. Females had higher GGT (median GGT value in females 4.0 vs 3.0 in males; P = 0.011) and albumin values (mean albumin value 3.3 in females vs 3.1 in males; P = 0.013). Conclusions and relevance Currently published RIs for some haematological and biochemical parameters are not appropriate for use in adult Maine Coon cats. A breed-specific variation could be a plausible explanation for the new haematological and serum biochemical analytes proposed in this study. Breed-specific RIs for Maine Coon cats will help prevent misinterpretation of laboratory results in diagnosis and in the selection of ideal blood donors.

Comparison of three blood transfusion guidelines applied to 31 feline donors to minimise the risk of transfusion-transmissible infections

Objectives The increased demand for animal blood transfusions creates the need for an adequate number of donors. At the same time, a high level of blood safety must be guaranteed and different guidelines (GLs) deal with this topic. The aim of this study was to evaluate the appropriateness of different GLs in preventing transfusion-transmissible infections (TTI) in Italian feline blood donors. Methods Blood samples were collected from 31 cats enrolled as blood donors by the owners’ voluntary choice over a period of approximately 1 year. Possible risk factors for TTI were recorded. Based on Italian, European and American GLs, specific TTI, including haemoplasmas, feline leukaemia virus (FeLV), feline immunodeficiency virus (FIV), Anaplasma phagocytophilum, Ehrlichia species, Bartonella species, Babesia species, Theileria species, Cytauxzoon species, Leishmania donovani sensu lato and feline coronavirus (FCoV), were screened. Rapid antigen and serological tests and biomolecular investigations (PCR) were used. Several PCR protocols for haemoplasma and FeLV DNA were compared. Results The presence of at least one recognised risk factor for TTI was reported in all cats. Results for FIV and FeLV infections were negative using rapid tests, whereas five (16.1%) cats were positive for FCoV antibodies. Four (12.9%) cats were PCR positive for haemoplasma DNA and one (3.2%) for FeLV provirus, the latter being positive only using the most sensitive PCR protocol applied. Other TTI were not detected using PCR. Conclusions and relevance Blood safety increases by combining the recommendations of different GLs. To reduce the risk of TTI, sensitive tests are needed and the choice of the best protocol is a critical step in improving blood safety. The cost and time of the screening procedures may be reduced if appropriate tests are selected. To this end, the GLs should include appropriate recruitment protocols and questionnaire-based risk profiles to identify suitable donors.

Transthoracic ultrasound guided balloon dilation of cor triatriatum dexter in 2 Rottweiler puppies

Balloon dilation was performed in two Rottweiler puppies with cor triatriatum dexter and clinical signs of ascites using transthoracic echocardiographic guidance. The dogs were positioned on a standard echocardiography table in right lateral recumbency, and guide wires and balloon catheters were imaged by echocardiographic views optimized to allow visualization of the defect. The procedures were performed successfully without complications and clinical signs were resolved completely in both cases. Guide wires and balloon catheters appeared hyperechoic on transthoracic echocardiography image and could be clearly monitored and guided in real-time. These two cases demonstrate that it is possible to perform balloon catheter dilation of cor triatriatum dexter under transthoracic guidance alone.