Detection of bacterial contamination and DNA quantification in stored blood units in 2 veterinary hospital blood banks.

Blood transfusions in veterinary medicine have become increasingly more common and are now an integral part of lifesaving and advanced treatment in small and large animals. Important risks associated with transfusion of blood products include the transmission of various infectious diseases. Several guidelines suggest what infectious agents to screen for in canine and feline transfusion medicine. However, while the risk of bacterial contamination of blood products during storage and administration has not been documented in veterinary medicine, it has emerged as a cause of morbidity and mortality in human transfusion medicine. Clinical experience shows that the majority of blood component bacterial contaminations are caused by only a few species. Unlike other types of bacteria, psychrotolerant species like Pseudomonas spp. and Serratia spp. can proliferate during the storage of blood units at 4°C from a very low titer at the time of blood collection to a clinically significant level (> 10(5) CFU/mL) causing clinical sepsis resulting from red blood cell concentrate transfusions in human medicine. The purpose of this report was to describe the detection and quantification procedures applied in 4 cases of bacterial contamination of canine and feline blood units, which suggest the need for further investigations to optimize patients' safety in veterinary transfusion medicine.

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, ß1 -globulin, ß2 -globulin, γ1 -globulin, and γ2 -globulin concentrations, and for the Albumin/Globulin ratio. Group S showed higher concentrations of TP, α2 -, ß1 -, ß2 -, 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.

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.

Prevalence and diagnosis of Babesia and Theileria infections in horses in Italy: a preliminary study.

Babesia caballi and Theileria equi are the causative agents of equine piroplasmosis. In this preliminary epidemiological study, 412 horses reared in central and northern Italy were sampled and three diagnostic methods compared, namely, the microscopy, the indirect fluorescent antibody test (IFAT) and a PCR. Possible risk factors (such as area, season, breed, activity, sex, age, and grazing) associated with serological positivity were evaluated. A seroprevalence of 68.4% was found: 12.4% of the animals had anti-T. equi antibodies, 17.9% anti-B. caballi antibodies and 38.1% had antibodies against both species. Of the seropositive samples, 3.1% and 9.4% were positive to microscopy and PCR, respectively; 31.5% of the horses were IFAT-negative but 1.4% and 2.4% of the corresponding blood samples were positive to microscopy and PCR, respectively. Molecular techniques revealed that the species present were closely related to T. equi, Theileria sergenti, Theileria buffeli and the Babesia microti-like piroplasm provisionally named Theileria annae. Grazing was found to be a pronounced risk factor for equine piroplasmosis.

Molecular characterisation of Babesia canis canis and Babesia canis vogeli from naturally infected European dogs.

The morphologically small Babesia species isolated from naturally infected dogs in Europe, Japan, and US are described as Babesia gibsoni despite the fact that molecular techniques show that they should be assigned to two or three separate taxons. The morphologically large Babesia isolated from dogs in Europe, Africa, and US were generally classified as B. canis until it was proposed to distinguish three related, albeit genetically distinct subspecies of this genus, namely B. canis canis, B. canis rossi, and B. canis vogeli. The insight into the molecular taxonomy of canine piroplasms is, however, limited because only partial small subunit ribosomal RNA (ssrRNA) sequence data exist for two species from the B. canis group. In this work, we molecularly characterised natural Babesia infections in 11 dogs from Croatia, France, Italy, and Poland. These infections were diagnosed as caused by B. canis canis and B. canis vogeli based on the analysis of the complete sequence of the ssrRNA genes. Phylogenetic analysis confirmed that the large Babesia species of dogs belong the to the Babesia sensu stricto clade, which includes species characterised by transovarial transmission in the tick vectors and by exclusive development inside the mammalian host erythrocytes. The new data facilitate the reliable molecular diagnosis of the subspecies of B. canis.