Search

Antimicrobial resistance is a widespread global health problem. The presence of resistant bacteria and antibiotic resistance genes has been demonstrated not only in humans but also in animals, including pets. Stray cats share the urban environment with people and pets. This may facilitate transmission of resistant bacteria and resistance genes between stray animals, people and domestic animals. Several studies have investigated the role of stray cats as a fecal carrier of ESBL-producing bacteria. However, there are many genes and resistance mechanisms that can be detected in commensal E. coli , which, because of its genetic plasticity, is considered an indicator for monitoring antibiotic resistance. In this study, rectal swabs were collected from stray cats from colonies and shelters in the city of Monza (Monza Brianza, Italy) to isolate commensal E. coli . Phenotypic tests, such as the minimum inhibitory concentration (MIC) and the double disc test (DDST), and molecular analyses to detect antimicrobial resistance genes (ARGs) were used to study the resistance of these isolates. The results obtained confirm that stray cats can carry ESBL-producing E. coli (6.7%) and genes conferring resistance to other important antibiotic classes such as tetracyclines and sulfonamides.

Feline leishmaniosis is a worldwide infection caused by the parasite of the genus Leishmania transmitted by sandflies. Based on the complexity of epidemiology and diagnosis of this infection, the role of cats in the epidemiology and clinical impact of disease is still under debate. By using serological and molecular methods, this study aimed to update the epidemiology of the infection in different feline populations from various areas of Italy and to study factors associated with the infection. Of 1490 cats tested, 124 (8.3%, 95% CI 6.9-9.9) were infected, 96 had only specific L. infantum IgG, 18 were only positive for parasite DNA and 10 were both IFAT and qPCR positive. Risk factors for infection were sampling in the winter season (OR = 3.2, 95% CI 2.2-4.8), originating from the Sicily region (OR = 2.0, 95% CI 1.3-3.0), male gender (OR = 1.8, 95% CI 1.1-3.2), outdoor lifestyle (OR = 2.3, 95% CI 0.9-5.6) and seropositivity for FIV antibodies (OR = 2.2, 95% CI 1.2-4.2), while sampling in the spring (OR = 0.5, 95% CI 0.3-0.7) and summer (OR = 0.3, 95% CI 0.1-0.7), and originating from the Lazio region (OR = 0.1, 95% CI 0.05-0.4) were protective factors for infection. In endemic areas, Leishmania infection should be investigated by using both serological and molecular methods and cats should be protected from sandfly bites, particularly if they are FIV infected.

Background: In humans, blood groups are associated with varying prevalence of infections. The aim of this study was to determine if associations exist between the feline AB blood group system and haemoplasma infection., Methods: Data from two studies were combined. In the first study, DNA samples from 131 haemoplasma-infected and 132 haemoplasma-uninfected UK cats underwent pyrosequencing to determine their blood genotype as AA, Ab or bb. In the second study, blood samples from 160 Italian cats of known blood phenotype A, B or AB underwent PCR testing for feline haemoplasma species DNA., Results: Haemoplasma infection was demonstrated in cats of all phenotypes and genotypes. A significantly higher number of Ab genotype cats tested positive for overall haemoplasma infection status (p = 0.04) and for Mycoplasma haemofelis infection (p = 0.03)., Limitations: Haemoplasma-infected Italian cats were few, possibly increasing the chance of type II error, and the presence of purebred cats in the sample population may have had a confounding effect., Conclusions: Feline haemoplasmas do not appear to preferentially use either blood type A or B antigens as attachment sites for erythrocyte colonisation. Further investigations in a larger number of haemoplasma-infected cats of known blood phenotype are warranted to explain the association between genotype Ab and haemoplasma infection., (© 2022 The Authors. Veterinary Record published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

Cats are susceptible to coronavirus infections, including infection by human severe acute respiratory syndrome coronavirus (SARS-CoV). In human ABO system blood groups, alloantibodies can play a direct role in resistance to infectious diseases. Individuals with the AB blood type were over-represented in the SARS-CoV-2 infection group. Blood type AB individuals lack both anti-A and anti-B antibodies, and therefore lack the protective effect against SARS-CoV-2 infection given by these antibodies. Starting from this knowledge, this pilot preliminary study evaluated a possible association between feline blood phenotypes A, B, and AB and serostatus for SARS-CoV-2 antibodies in cats. We also investigated selected risk or protective factors associated with seropositivity for this coronavirus. A feline population of 215 cats was analysed for AB group system blood phenotypes and antibodies against the nucleocapsid (N-protein) SARS-CoV-2 antigen using a double antigen ELISA. SARS-CoV-2 seropositive samples were confirmed using a surrogate virus neutralization test (sVNT). Origin (stray colony/shelter/owned cat), breed (DSH/non DSH), gender (male/female), reproductive status (neutered/intact), age class (kitten/young adult/mature adult/senior), retroviruses status (seropositive/seronegative), and blood phenotype (A, B, and AB) were evaluated as protective or risk factors for SARS-CoV-2 seropositivity. Seropositivity for antibodies against the SARS-CoV-2 N-protein was recorded in eight cats, but only four of these tested positive with sVNT. Of these four SARS-CoV-2 seropositive cats, three were blood phenotype A and one was phenotype AB. Young adult age (1-6 years), FeLV seropositivity and blood type AB were significantly associated with SARS-CoV-2 seropositivity according to a univariate analysis, but only blood type AB ( p = 0.0344, OR = 15.4, 95%CI: 1.22-194.39) and FeLV seropositivity ( p = 0.0444, OR = 13.2, 95%CI: 1.06-163.63) were significant associated risk factors according to a logistic regression. Blood phenotype AB might be associated with seropositivity for SARS-CoV-2 antibodies. This could be due, as in people, to the protective effect of naturally occurring alloantibodies to blood type antigens which are lacking in type AB cats. The results of this pilot study should be considered very preliminary, and we suggest the need for further research to assess this potential relationship.

The emergence and spread of antimicrobial resistance (AMR) is a global problem that requires a One Health approach. Despite several studies have reported the role of companion animals as reservoirs of AMR, limited information is available regarding the role of cats in the circulation of AMR. In this study, we evaluated the phenotypic and genotypic profile of 75 Escherichia coli isolated from rectal swabs and fecal samples of 75 stray cats (living in solitary or in a colony) sampled in Palermo (Sicily, Italy), to determine whether these animals may participate in the spread of AMR. Susceptibility to 8 antibiotics was tested using Minimum Inhibitory Concentration assays, while the presence of the common antibiotic resistance genes bla TEM , bla CTX-M , tet (A), and tet (B) was investigated by PCR. From the 75 E. coli isolates analyzed, 43% were resistant to at least one of the eight antibiotics tested, with 31% of the isolates resistant to ampicillin, 23% to cefotaxime, 21% to tetracycline, 20% to cefazolin, and 17% to amoxicillin/clavulanic acid. Most isolates harbored the bla TEM gene (29%), followed by bla CTX-M (23%), tet (A) (21%), and tet (B) (20%). Our results confirm the fecal carriage of antibiotic-resistant E. coli and clinically relevant resistance genes in stray cats. This study highlights the potential role of stray cats in the spread of AMR in urban environments, emphasising the need to better understand their role in AMR circulation when planning strategies to combat it.

Low-level laser therapy (LLLT) is a therapeutic option that stimulates cellular function through intracellular photobiological and photochemical reactions, promoting better tissue repair and an anti-inflammatory, antibacterial, and analgesic effect. Previous studies in human and veterinary medicine have shown the clinical efficacy of LLLT in many fields. In this study, the literature was reviewed using the critically appraised topic (CAT) method to determine the canine skin diseases for which LLLT is an advisable treatment. A meticulous literature search revealed 19 significant clinical trials, which were critically analyzed. The evaluation of the best accessible evidence in July 2022 suggests that fluorescence biomodulation (FBM), a type of LLLT, can, in combination with systemic antibiotic therapy, be a promising and effective adjunctive treatment for canine interdigital pyoderma and canine deep pyoderma. Furthermore, the evidence suggests that the use of LLLT is not recommended as a therapy for pedal pruritus secondary to canine atopic dermatitis. For other canine skin diseases included in the CAT, although LLLT appears to be a promising treatment, there is not yet good scientific evidence to recommend its use.

The leishmaniases are vector-borne parasitic diseases affecting humans and animals, with high mortality rates in endemic countries. Infected dogs represent the main reservoir of infection. Disease control is mainly based on chemotherapy, which, at present, shows serious drawbacks both in humans and dogs. Therefore, the discovery or repurposing of new treatments is mandatory. Here, three monovalent ionophores (salinomycin, monensin, nigericin) were tested against promastigotes of Leishmania (L.) infantum, Leishmania tropica, and Leishmania braziliensis, and against amastigotes of L. infantum within human and, for the first time, canine macrophages. All three drugs were leishmanicidal against all Leishmania spp. promastigotes with IC50 values between 7.98 and 0.23 µM. Monensin and nigericin showed IC50 values < 1 µM, whereas salinomycin was the least active compound (IC50 > 4 µM). Notably, the ionophores killed L. infantum amastigotes within human THP-1 cells with IC50 values ranging from 1.67 to 1.93 µM, but they only reduced by 27−37% the parasite burden in L. infantum-infected canine macrophages, showing a host-specific efficacy. Moreover, a selective higher toxicity against canine macrophages was observed. Overall, repurposed ionophores have the potential to be further investigated as anti-Leishmania agents, but different drug options may be required to tackle human or canine leishmaniases.

Cats are susceptible to feline coronavirus (FCoV), a highly contagious virus with fecal-oral transmission. In people, susceptibility to coronavirus infection, such as SARS-CoV infection, has been associated with the ABO blood group, with individuals with blood group O having significantly lower risk of SARS-CoV infection. This study evaluated a possible association between feline blood group phenotypes A, B and AB and serostatus for antibodies against FCoV. We also investigated risk or protective factors associated with seropositivity for FCoV in the investigated population. Feline populations were surveyed for AB group system blood types and for presence of antibodies against FCoV. Blood phenotype, origin, breed, gender, reproductive status and age of cats were evaluated as protective or risk factors for coronavirus infection. No blood type was associated with FCoV seropositivity, for which being a colony stray cat ( p = 0.0002, OR = 0.2, 95% CI: 0.14-0.54) or a domestic shorthair cat ( p = 0.0075, OR = 0.2, 95% CI = 0.09-0.69) were protective factors. Based on results of this study, feline blood phenotypes A, B or AB do not seem to predispose cats to seropositivity for FCoV. Future studies on other feline blood types and other infections could clarify whether feline blood types could play a role in predisposing to, or protecting against, feline infections.

Objectives: The relationship between blood group antigens and disease has been studied in humans. Blood types have been associated with both decreased and increased rates of various infections. In addition, blood group expression has been shown to vary with some cancers and gastrointestinal diseases. The objective of this study was to explore whether there is a relationship between blood type and retroviral infections in cats., Methods: Case records from a veterinary research laboratory, veterinary teaching hospitals and veterinary blood banks were retrospectively searched for cats where both blood type and retroviral status (feline leukemia [FeLV], feline immunodeficiency virus [FIV] or both) were listed (part 1). In addition, a sample of 33 cats with confirmed FIV infection was genotyped to determine blood groups (part 2)., Results: In part 1, 709 cats were identified, 119 of which were positive for retroviral infection. Among all cases, 621 were type A (87.6%), 68 were type B (9.6%) and 20 were type AB (2.8%). There was no relationship between overall retroviral status (positive/negative) and blood type ( P  = 0.43), between FeLV status and blood type ( P  = 0.86) or between FIV status and blood type ( P  = 0.94). There was no difference in the distribution of blood types between cats that were healthy and typed as possible blood donors vs sick cats that were typed prior to a possible transfusion ( P  = 0.13). In part 2, of the 33 FIV-infected cats, all blood group genotypes were identified, although this test did not discriminate type A from type AB., Conclusions and Relevance: No relationship was identified between feline retroviral status and blood type in this study. The relationship between blood type and other disease states requires further study in veterinary patients.

This study measured the changes of hemostatic activity in liquid plasma (LP) over 7 days of storage. Five canine plasma units, divided into two aliquots were evaluated: one stored refrigerated at 2-6°C as never-frozen LP and one frozen at -18°C as fresh frozen plasma (FFP). Clotting times, coagulation activities of factor (F) V, VIII, X, XI, antithrombin (AT), and von Willebrand (vWF), fibrinogen and D-dimers (DD) content were assessed before storage (baseline value), and after 12, 24, 48 h and 7 days (D7) in LP stored refrigerated, and on day 7 in FFP. At baseline median values of all factor activity were greater than 80%, and for clotting times, AT, fibrinogen and DD content, were within the canine reference range. Some hemostatic parameters changed significantly over 7 days and at the end of storage in LP. However, median activities of FV, FVIII, FX and FXI, coagulation time, AT, fibrinogen and DD content remained within reference ranges at all time points. The only exception was for vWF which median activity was lower than reference range for all storage time points. Activity of FVIII was significant lower in LP at D7 when compared to activity in FFP, with values of 62 vs. 118%, respectively. DD content showed a median value higher than reference range in FFP at D7. Despite some statistically significant changes at the end of 7-day storage period, never-frozen LP maintained median factor activities >80% for most factors. The clinical impact of the drop over time of vWF activity is unknown., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Proverbio, Perego, Baggiani and Spada.)

The tiger ( Panthera tigris ) is an endangered species. The health of individuals is important and any data on hematological and biochemical blood values can provide valuable information; when combined with physical assessment. This data assists in both the diagnosis of disease and some conservation strategies. The behavior of wild tigers makes it is extremely difficult to obtain biological samples from free-living subjects, therefore, data collected from captive tigers is highly valuable. The aim of this study was to provide additional information for the values of hematological and serum biochemical parameters in healthy captive tigers. Blood samples were collected from 22 clinically healthy tigers ( Panthera tigris) . The following parameters were analyzed: glucose, urea, creatinine, alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein (TP) and red blood cells (RBCs), hemoglobin (Hb), hematocrit (Hct) and red cell indices; such as mean cell volume (MCV), mean cell Hb (MCH), mean cell Hb concentration (MCHC), platelet (PLT) and white blood cells (WBCs). The mean hematological values in our tiger population were not significantly different when compared with the same parameters in the previously studied tiger population. The mean values of RBCs and PLT were statistically significantly higher and the mean values of Hb, PCV, MCV, MCH, MCHC, and WBC were lower than the mean values obtained in previous studies on the Amur tiger. Further investigation of captive and free-living tigers is needed to identify the normal ranges for parameters in this endangered species.

Leukocyte- and platelet-rich plasma (L-PRP) can accelerate the healing process by providing increased concentrations of platelet-derived growth factors. The objective of this study was to evaluate the clinical effect of L-PRP in the treatment of canine aural hematomas associated with otitis externa. Twenty mL of citrated whole blood was collected from each of the 17 dogs included and autologous L-PRP was produced. The aural hematoma was drained and 0.5-1 mL of L-PRP was injected. The dogs were examined weekly until 7 days after complete clinical healing. A final clinical follow-up was performed 6 weeks after the first treatment with L-PRP. If there was recurrence of the aural hematoma at the first follow-up, the treatment was repeated. In total, 2/17 cases were lost after the first follow-up. In 5/17 dogs, a short-term recurrence occurred. In 12/15 cases, complete clinical resolution was achieved with a single L-PRP application (Group A1) and in 3/15 with two treatments (Group A2). The mean time to complete clinical resolution was 16 ± 8.7 days (A1) and 23.3 ± 4 days (A2), respectively. No side effects were reported. The in situ administration of autologous L-PRP resulted in a complete resolution of the aural hematoma in all dogs that completed the clinical trial.

Leukoreduction of blood products is a technique used to prevent leukocyte-induced transfusion reactions and is extensively used in human, but rarely in veterinary patients. The concentration of some coagulation proteins can be affected by the processing steps used for the preparation of leuko-reduced plasma units. In this study, we assessed the effect of leukoreduction on coagulation activity of canine plasma collected for transfusion. Ten plasma units, five obtained from non-leuko-reduced (non-LR) whole blood (WB) units and five from leuko-reduced (LR) WB units were evaluated. Prothrombin time (PT), activated partial thromboplastin time (aPTT), coagulation factor activities of factors (F) V, VIII, X, XI, and von Willebrand (vWF), fibrinogen and D-dimers content were assessed at collection (baseline value, D0) and after 7 days of frozen storage at -18 °C (D7). Compared to non-LR plasma units, LR units showed a statistically significant prolonged aPTT and reduced FXI activity. Filtration had no significant effect on the other factors and parameters evaluated. Filtration-dependent changes appear to have no impact on the therapeutic quality of plasma obtained from leuko-reduced whole blood, other than for FXI activity. Further studies on a larger sample size comparing the same unit before and after leukoreduction are needed to confirm these findings.

Cats are susceptible to infection with severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2). Whilst a number of studies have been performed worldwide on owned cats, limited data are available on stray, colony or shelter cats. We investigated SARS-CoV-2 infection in a stray cat population before and during human outbreaks of SARS-CoV-2 in cities in the Lombardy region in northern Italy, a high endemic region for SARS-CoV-2, using serological and molecular methods. A cohort of different samples were collected from 241 cats, including frozen archived serum samples from 136 cats collected before the 2019 coronavirus disease (COVID-19) pandemic and serum, pharyngeal and rectal swab samples from 105 cats collected during the SARS-CoV-2 outbreak. All pre-pandemic samples tested seronegative for antibodies against the nucleocapsid of SARS-CoV-2 using indirect enzyme linked immunosorbent assay (ELISA) test, while one serum sample collected during the pandemic was seropositive. No serological cross-reactivity was detected between SARS-CoV-2 antibodies and antibodies against feline enteric (FECV) and infectious peritonitis coronavirus (FIPC), Feline Immunodeficiency Virus (FIV), Feline Calicivirus (FCV), Feline Herpesvirus-1 (FHV-1), Feline Parvovirus (FPV), Leishmania infantum , Anaplasma phagocytophilum , Rickettsia spp., Toxoplasma gondii or Chlamydophila felis . No pharyngeal or rectal swab tested positive for SARS-CoV-2 RNA on real time reverse transcription-polymerase chain reaction (rRT-PCR). Our data show that SARS-CoV-2 did infect stray cats in Lombardy during the COVID-19 pandemic, but with lower prevalence than found in owned cats. This should alleviate public concerns about stray cats acting as SARS-CoV-2 carriers.

Background: To characterize the cellular composition (platelets, erythrocytes, and leukocytes) and determine platelet-derived growth factor isoform BB (PDGF-BB) concentration in canine leukocyte- and platelet rich plasma (L-PRP) produced using a commercial semi-automated closed system., Methods: Twenty milliliters of citrated whole blood were obtained from 30 healthy un-sedated canine blood donors and processed using a semi-automated completely closed commercial system (CPUNT 20, Eltek group, Casale Monferrato, Alessandria, Italy) according to the manufacturer's instructions. Erythrocyte, leukocyte, and platelet counts were determined in both whole blood (WB) and resultant L-PRP. The PDGF-BB concentration was evaluated after bovine thrombin activation of 10 L-PRP samples., Results: This commercial system produced on average 2.3 ± 0.7 mL of L-PRP containing a high concentration of platelets (767,633 ± 291,001 μL, p < 0.001), with a 4.4 fold increase in platelet count, lower concentration of erythrocytes (528,600 ± 222,773 μL, p < 0.001) and similar concentration of leukocytes (8422 ± 6346 μL, p = 0.9918) compared with WB. L-PRP had an average of 3442 ± 2061 pg/mL of PDGF-BB after thrombin activation. Neutrophils, lymphocytes and monocytes average percent content in L-PRP was 14.8 ± 13.2, 71.7 ± 18.5 and 10.7 ± 6.4, respectively., Conclusion: Sterile canine L-PRP prepared using this semi-automated closed system is easy to obtain, produces a significant increase in platelet count compared to WB and contains a detectable concentration of PDGF-BB after activation. Additional in vitro and in vivo studies are needed to assess inflammatory markers concentration and the therapeutic efficacy of this L-PRP in dogs.

A knowledge of the blood groups and alloantibodies present is essential for the safe transfusion of blood products in horses. Pre-transfusion screening and blood typing minimizes the risk of incompatible RBC transfusions and prevents immunization of the recipient against incompatible RBC antigens. The frequencies of blood groups can vary among different breeds. Knowledge of a breed's blood group prevalence can be very useful for identifying the best blood donors during transfusion in clinical practice. The aims of this study were to estimate the prevalence of the Ca blood type in horses from Italy using a monoclonal immunocromatographic method and to estimate the prevalence of anti-Ca alloantibodies in Ca- horses using agglutination on gel technique. Ca blood type was determined on 110 whole blood samples. The prevalence of the Ca+ blood type was 79.1%. This study also provides data about the prevalence of Ca+ blood group in Italian Saddle Horses (77,3%) and Dutch Warmblood (58,3%). No significant association was found between Ca blood type and sex with 79.5% and 78.8% of females and males testing Ca+, respectively. The total number of Ca- samples with detectable anti-Ca alloantibodies was 7/23 (30.4%).

The aims of this study were to determine the prevalence of A, B and AB blood types and alloantibodies in non-pedigree cats from two regions, one in Northern and one in Southern Italy (Lombardy and Sicily, respectively). A total of 448 samples (52.0% from Northern and 48.0% from Southern Italy) were blood typed. The prevalence of A, B and AB blood types in northern and southern cats were 91.0%, 5.2%, 3.8%, and 77.2%, 12.1% and 10.7%, respectively. The prevalence of type-A blood in southern cats was significantly lower ( p = 0.0001) than in northern cats, while type-B and AB blood were significantly higher ( p = 0.0085 and p = 0.0051, respectively) in Southern compared to Northern Italian cats. Alloantibodies against type-A blood were found in 94.1% of type-B cats, 11.2% of type-A cats had alloantibodies against type-B blood, while no type-AB cats had alloantibodies with no significant difference between the two Italian populations. Type-AB prevalence in non-pedigree cats in Southern Italy was the highest reported in Europe. Italian type-A cats had the lowest worldwide prevalence of alloantibodies against type-B blood. These results highlight the usefulness of regional studies to report different prevalences in feline blood types and reinforce the importance of blood typing cats before transfusions and mating.

Gel technology is widely used for blood typing in human medicine. It has a number of advantages over routine tube testing, including standardization, stability, smaller sample volume, ease of performance and analysis, and speed. The aim of this study was to evaluate feline blood typing using the gel column technique. TUBE agglutination typing was performed in 143 feline blood samples from blood donors and recipients, healthy and sick patients, and whole-blood units anticoagulated with ethylenediamine tetraacetic acid or citrate phosphate dextrose adenine. Plasma from type B cats was used as anti-A reagent, Triticum vulgaris lectin as anti-B reagent, and the control was saline solution. Agglutination in backtyping of types B and AB samples with type A red blood cells (RBCs) was used to confirm whether the samples were type B (presence of alloantibodies) or type AB (absence of alloantibodies). Blood typing in a neutral gel column technique (GEL) using the same anti-A and anti-B reagents was performed on duplicate samples. Sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, negative predictive value, overall accuracy, and Cohen κ coefficient (κ) for GEL were calculated, with TUBE considered the gold standard technique. Of 143 samples typed with TUBE, 98 (68.5%) were type A, 25 (17.5%,) type B, and 20 (14.0%) type AB. Backtyping confirmed the categorization of all types B and AB samples. Of these samples, gel testing produced 115 (80.4%) concordant results; a mixed-field agglutination pattern (layers of RBCs at both the top and at the bottom of the gel in either the A or B gel column) was seen in 27 samples, and one type B sample was misidentified as type AB. If the mixed-field pattern was interpreted as a negative result, 141/143 (98.6%) samples showed concordant results with an overall accuracy of the GEL of 100.0% for type A, 98.9% for type B, and 99.1% for type AB. Strength of agreement was very good (κ = 0.97). When the same anti-A and anti-B reagents are used, GEL is a sensitive and specific method for blood typing feline samples. Until additional studies have been performed, mixed-field patterns obtained in GEL testing should be classified as negative results., (Copyright © 2020 Spada, Perego, Baggiani and Proverbio.)

Feline leishmaniosis (FeL) is an emerging vector-borne feline disease, with increasing numbers of cases reported and studies performed internationally. This study aimed to update the epidemiological status for FeL in stray cats in Milan, northern Italy; compare these results with previous studies in Northern Italy; and report clinicopathologic findings and coinfections in cats infected with Leishmania spp. A total of 117 cats were tested for L. infantum and retrovirus infection, hematological, and biochemical parameters. Demographic and clinical data were collected and FeL affected cats screened for selected coinfections. Overall, 10/117 (8.6%) cats tested positive for L. infa n tum : in five cats L. infantum DNA was found in popliteal lymph nodes and five were IFAT seropositive at titers from 1:80 to 1:160. Infected cats were concentrated in a specific area of Milan ( p = 0.0154). No specific clinicopathologic abnormalities or retroviral infections were significantly linked to the infection, other than hypergammaglobulinemia ( p = 0.0127). Seroreactivity to Anaplasma phagocytophilum , Chlamydophila felis , and Toxoplasma gondii was found in some infected cats. A high prevalence of FeL was found in a non-endemic area of northern Italy and future studies should continually monitor this data to understand whether these cases are imported or if Leishmania vectors are present in this area., Competing Interests: The authors declare no conflict of interest.

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.

Given the endangered status of tigers ( Panthera tigris ), the health of each individual is important and any data on blood chemistry values can provide valuable information alongside the assessment of physical condition. The nature of tigers in the wild makes it is extremely difficult to obtain biological samples from free-living subjects, therefore the values obtained from captive tigers provide very useful data. Serum protein electrophoresis is a useful tool in the diagnosis and monitoring of a number of diseases. In this study, we evaluated agarose gel serum protein electrophoresis on samples from 11 healthy captive tigers. Serum electrophoresis on all 11 tiger samples successfully separated proteins into albumin, α 1 , α 2 , β 1 , β 2 and γ globulin fractions as in other mammals. Electrophoretic patterns were comparable in all tigers. Mean± standard deviation or median and range values obtained for each protein fraction in healthy tigers were, respectively: 3.6 ± 0.2, 0.21 (0.2-0.23), 1.2 ± 0.2, 10.7 ± 0.2, 0.4 (0.3-0.6), 1.2 (1-1.8) gr/dL. The results of this preliminary study provide the first data on serum electrophoretic patterns in tigers and may be a useful diagnostic tool in the health assessment of this endangered species.

Background and Aim: Empirical antimicrobial therapy is frequently given in superficial bacterial folliculitis (SBF) and otitis externa (OE) in dogs, especially for the initial clinical presentation. Culture and subsequent antimicrobial susceptibility testing (AST) are generally limited to chronic cases with poor response to initial therapy. Several factors contribute to the failure to implement the use of AST in veterinary practice, i.e., long laboratory turnaround time or special requirements for sample shipping. Point-of-care (PoC) testing might reduce laboratory turnaround time and costs and the risk of emergence of multidrug-resistant pathogens. This study evaluated the Speed Biogram™ PoC test in canine SBF and OE compared with conventional methods for culture and AST., Materials and Methods: Thirty-four canine samples were analyzed: eleven from SBF, seven from bacterial OE, four from mixed OE, six from Malassezia spp. OE, and six negative controls. Sensitivity (Se) and specificity (Sp) of the PoC test and the agreement between the PoC test and conventional methods were evaluated., Results: Se and Sp of PoC test in discriminating between healthy and unhealthy subjects were 100% (95% confidence interval [CI] 87.66-100.00) and 100% (95% CI 54.1-100.0), respectively. For bacterial identification, the k value was 0.532. Se and Sp of PoC tests for AST were 81.73% (95% CI 72.95-88.63) and 93.10% (95% CI 88.86-96.98), respectively with a total good agreement between tests (mean k=0.714), but major (8/27) and very major (19/27) errors were observed in 55% of bacterial conventional culture-positive samples., Conclusion: PoC test can identify dogs with SBF and OE, but AST is not sufficiently accurate. The lack of susceptibility testing for methicillin makes this test inappropriate for use in small animal practice., (Copyright: © Perego, et al.)

Background: The appropriate cutoff to define a positive point-of-care card agglutination (CA) test for dog erythrocyte antigen 1 (DEA 1) blood typing depends on whether the test can be used in the donor or recipient., Objectives: By screening for CA test positivity, we aimed to evaluate the best cutoff value for DEA 1 blood typing in canine blood donors using a receiver operator characteristic (ROC) curve., Methods: Ethylenediaminetetraacetic acid (EDTA) blood samples from 100 canine blood donors were blood-typed in parallel for DEA 1 using both immunochromatographic (IC) and CA tests. The effect of temperature, storage time, and anticoagulant solutions for both methods was evaluated. Unweighted and weighted Cohen's Kappa (K) statistic was calculated to evaluate the agreement between the two testing methods. The overall performance of the CA test was evaluated by generating a ROC curve using the IC test as the reference method., Results: Concordant results were obtained for 86% of the samples. Unweighted and weighted K statistics demonstrated good and moderate agreement, respectively. For the CA test, the ROC curve showed an area under the curve (AUC) of 0.910, with the highest sensitivity cutoff values at ≥1+ agglutination. CA- and IC-typed EDTA blood samples stored at room temperature for up to 1 week and refrigerated for up to 1 month were concordant as were the citrate phosphate dextrose adenine 1 (CPDA-1) anticoagulated blood samples stored for up to 1 week at 4 ± 2°C., Conclusions: The overall reliability of the CA method seemed to be lower than that of the IC method. When CA is used as a screening test for canine blood donors, the correct cut off is ≥1+ agglutination is recommended to maximize sensitivity., (© 2019 American Society for Veterinary Clinical Pathology.)

Objectives: Despite the increasing availability of feline blood collected and stored for transfusion purposes, few studies have been performed on feline blood units. The aim of this prospective in vitro study was to evaluate haematological and morphological changes in feline blood cells in whole blood units between collection and end of storage., Methods: Haematological examination (red blood cells [RBCs], haemoglobin, haematocrit, red cell distribution width, mean cell volume, mean cell haemoglobin concentration, mean cell haemoglobin, white blood cells [WBCs] and platelet [PLT] count) was performed on 40 non-leukoreduced feline whole blood units at the time of collection (day[D]0) and after storage (D35). The blood was collected into citrate-phosphate-dextrose-adenine anticoagulant-preservative solution using an open system in a veterinary blood bank and stored for 35 days at 4 ± 2°C. Twenty of these feline whole blood units were also analysed for blood cell morphology (normal RBCs, macrocytes, echinocytes, spherocytes, schistocytes, lysed RBCs, RBCs with Heinz bodies and recognisable WBC and PLT count). Differences between the two examination times were statistically analysed., Results: There was a statistically significant decrease in WBC and PLT counts after storage at D35 ( P <0.0001 for both). The most significant cellular morphological changes after storage were an increase in echinocyte count ( P = 0.0001), and lysed RBCs ( P <0.0001), and a decrease in normal RBCs ( P <0.0001). Recognisable WBCs - mainly lymphocytes - were present at the end of storage., Conclusions and Relevance: This study showed that significant morphological changes occur in RBCs in feline blood units during storage for 35 days. In vivo studies are required to establish if these changes could affect the ability of stored RBCs to circulate and provide adequate oxygen delivery after transfusion.

Background: Canine generalised demodicosis is an inflammatory parasitic skin disease caused by an excessive proliferation of Demodex spp. Generalized demodicosis is a severe skin disease, that can be life threatening if not treated properly. Many of the current treatment options are not licensed for the treatment of generalised demodicosis, it have a low safety margin and may be poorly efficacious and time-consuming for the owner; there is a need for a safe, efficacious treatment for canine demodicosis. Our objective was to systematically review the literature to determine the most effective and safe topical or systemic therapy for canine generalised demodicosis. Single case reports and case series with fewer than five patients were not reviewed as they were considered to be poor quality evidence. A detailed literature search identified 21 relevant clinical trials and these were critically assessed., Results: The analysis of the best available evidence on March 5, 2018, suggests that six are the most effective and safe treatments for generalised canine demodicosis including (in alphabetical order): doramectin (oral or parenteral); fluralaner (oral); imidacloprid/moxidectin (topical); ivermectin (oral, not as first choice treatment); milbemycin oxime (oral); and sarolaner (oral). There was insufficient evidence to allow comment on the appropriateness of other treatment protocols for canine generalised demodicosis in this CAT., Conclusions: In our critical appraisal of the best scientific literature, there is evidence for recommending the use of 6 therapeutic options against demodectic mange. Further, in vivo, controlled, randomized and blinded clinical trials are required, to evaluate new therapies.

We compared 3 major cross-match (XM) tests to identify dog erythrocyte antigen (DEA) 7 blood incompatibilities in dogs as a result of anti-DEA 7 antibodies: gel (GEL), standard tube (TUBE) agglutination, and immunochromatography strips (STRIP). Blood samples from 42 dogs were typed for DEA 7; 2 tested DEA 7-positive (DEA 7+). The 40 DEA 7-negative (DEA 7-) plasma samples were cross-matched against the 2 DEA 7+ and 3 DEA 7- red blood cell (RBC) samples by GEL to identify samples with anti-DEA 7 antibodies. Twenty DEA 7- plasma samples without and with anti-DEA 7 antibodies were cross-matched with samples of the 2 DEA 7+ RBCs in a double-blind fashion using the TUBE and STRIP XM methods. GEL results were used as the reference method for comparison. To determine relationships between results, 2 × 2 tables were used. Cohen kappa coefficient (κ) was calculated between results of GEL and the other 2 methods. With GEL, 21 of 40 XM tests were positive and 19 of 40 negative for anti-DEA 7 antibodies. The same results were obtained by TUBE, whereas only 1 of 40 XM tests was positive by STRIP. There was a statistically significant relationship between results of GEL and TUBE ( p < 0.000) with perfect agreement (κ = 1.000), but not between GEL and STRIP results ( p = 1.000) in which agreement was equivalent to chance (κ = 0.0453). The GEL and TUBE XM tests, but not STRIP, are useful methods for identification of DEA 7 incompatibilities caused by anti-DEA 7 antibodies.

Objectives The aim of this study was to investigate the presence of naturally occurring antibodies against canine erythrocyte antigens in cats and vice versa. The influence of canine and feline blood type on cross-match results was also studied. Methods Blood samples from 34 cats and 42 dogs were used to perform test tube major and minor cross-match tests and blood typing. Blood from each cat was cross-matched with blood from 2-6 dogs, for a total of 111 cross-match tests. Haemolysis, macro- and microagglutination were considered markers of a positive cross-match. Results Eighty-three overall major cross-match tests were positive at 37°C, 86 at room temperature and 90 at 4°C. The minor cross-match tests were positive in all but two cross-matches performed at 37°C, all tests performed at room temperature and all but one test performed at 4°C. No cats tested totally negative at both major and minor cross-matches performed with samples from any single dog. Prevalence of warm natural antibodies against canine erythrocyte antigens was lower in type B cats than in type A cats, regardless of the blood type of donor dogs. Conclusions and relevance This study reveals a high prevalence of naturally occurring antibodies in cats against dog erythrocyte antigens and vice versa, and suggests that transfusion of cats with canine blood is not recommended as a routine procedure owing to the potential high risk of either acute severe or milder transfusion reactions.

[This corrects the article DOI: 10.1371/journal.pone.0154973.].

Feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) are among the most important feline infectious diseases worldwide. This retrospective study investigated survival times and effects of selected predictor factors on survival time in a population of owned pet cats in Northern Italy testing positive for the presence of FIV antibodies and FeLV antigen. One hundred and three retrovirus-seropositive cats, 53 FIV-seropositive cats, 40 FeLV-seropositive cats, and 10 FIV+FeLV-seropositive cats were included in the study. A population of 103 retrovirus-seronegative age and sex-matched cats was selected. Survival time was calculated and compared between retrovirus-seronegative, FIV, FeLV and FIV+FeLV-seropositive cats using Kaplan-Meier survival analysis. Cox proportional-hazards regression analysis was used to study the effect of selected predictor factors (male gender, peripheral blood cytopenia as reduced red blood cells - RBC- count, leukopenia, neutropenia and lymphopenia, hypercreatininemia and reduced albumin to globulin ratio) on survival time in retrovirus-seropositive populations. Median survival times for seronegative cats, FIV, FeLV and FIV+FeLV-seropositive cats were 3960, 2040, 714 and 77days, respectively. Compared to retrovirus-seronegative cats median survival time was significantly lower (P<0.000) in FeLV and FIV+FeLV-seropositive cats. Median survival time in FeLV and FIV+FeLV-seropositive cats was also significant lower (P<0.000) when compared to FIV-seropositive cats. Hazard ratio of death in FeLV and FIV+FeLV-seropositive cats being respectively 3.4 and 7.4 times higher, in comparison to seronegative cats and 2.3 and 4.8 times higher in FeLV and FIV+FeLV-seropositive cats as compared to FIV-seropositive cats. A Cox proportional-hazards regression analysis showed that FIV and FeLV-seropositive cats with reduced RBC counts at time of diagnosis of seropositivity had significantly shorter survival times when compared to FIV and FeLV-seropositive cats with normal RBC counts at diagnosis. In summary, FIV-seropositive status did not significantly affect longevity of cats in this study, unlike FeLV and FIV+FeLV-seropositivity. Reduced RBC counts at time of FIV and FeLV diagnosis could impact negatively on the longevity of seropositive cats and therefore blood counts should always be evaluated at diagnosis and follow-up of retrovirus-seropositive cats., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objectives The quality of whole blood (WB) units is influenced by many factors, starting with selection of donors and the method of blood collection. The aim of this study was to investigate the changes that occur in haematological and selected biochemical parameters in blood transferred from a feline blood donor to feline WB unit. Methods Data from 27 feline blood donations were used in this study. Cats were anaesthetised with a combination of tiletamine and zolazepam. Blood (10 ml/kg body weight to a maximum of 60 ml/cat) was collected in citrate-phosphate-dextrose-adenine (CPDA) anticoagulant. Lactated Ringer's solution (10 ml/kg) was administered intravenously starting halfway through the donation. Haematological and selected biochemical parameters (complete blood count, free haemoglobin, % haemolysis, glucose, sodium, potassium, pH) were measured in the blood donor before donation and in the corresponding donated WB unit soon after collection. Results Significant decreases occurred between blood donor and WB unit in red blood cells (mean difference -1.06 × 10 12 /l; P <0.0001), haemoglobin (mean difference -1.6 g/dl; P <0.0001), haematocrit (mean difference -4.6%; P <0.0001), red cell distribution width (mean difference -0.9%; P = 0.0003), white blood cells (mean difference -2.17 × 10 9 /l; P <0.0001), pH (mean difference -0.5; P <0.0001) and potassium (mean difference -1.4 mmol/l; P <0.0001). Significant increases occurred between blood donor and WB unit in platelets (mean difference +87.00 ×10 9 /l; P = 0.0039), glucose (mean difference +25.42 mmol/l; P <0.0001) and sodium (mean difference +20 mmol/l; P <0.0001). Conclusions and relevance When using a blood collection protocol with intravenous fluid administration midway through the donation and a CPDA:blood ratio of 1:7, there were significant changes in both the haematological and biochemical characteristics between the blood donors and WB units. The majority of these changes may be the result of the anticoagulants used for storage. Understanding these changes may assist selection of blood donors and help prediction of the characteristics of the donated WB unit.

Background: Identification of Leishmania infantum-infected dogs is crucial for control of canine leishmaniosis. In particular, in areas where access to specialized laboratories is limited, the availability of reliable and rapid in-clinic serologic tests may support immediate diagnosis in suspected cases and permit detection of asymptomatic canine carriers of L infantum infection., Objective: The purpose of the study was to validate the immunochromatographic test (ICT) Anigen Rapid Leishmania Ab Test kit for detection of L infantum antibodies in naturally exposed dogs in comparison with the immunofluorescence antibody test (IFAT)., Methods: Serum samples from 66 dogs, including 20 healthy control dogs and 46 dogs suspected or confirmed with canine leishmaniosis, were measured by both tests. Anti-Leishmania IgG titers ≥ 1:40 by IFAT were considered positive. Kappa statistic with a 95% CI was calculated to evaluate agreement between the 2 testing methods, and sensitivity, specificity, and positive and negative likelihood ratio were calculated., Results: Anti-L infantum IgG antibodies were found in 35 of 66 samples using the IFAT test (titers 1:40-1:5120). Thirty-one out of 66 samples tested positive with the qualitative ICT. Four IFAT-positive (titers < 1:80) samples were ICT-negative. The Kappa value of 0.853 demonstrated very good agreement between the 2 tests., Conclusion: The Anigen Rapid Leishmania Ab Test kit reliably identified canine sera with anti-L infantum IgG antibody titers ≥ 1:40. The ICT requires neither special preparation of the serum nor specialized equipment and can be stored at ambient temperature. The test is applicable as a field test because it is easy to use and provides rapid results., (© 2016 American Society for Veterinary Clinical Pathology.)

The reported prevalence of naturally occurring anti-dog erythrocyte antigen (DEA) 7 antibodies in DEA 7-negative dogs is as high as 50%. Characterization of these antibodies may better define their importance in canine transfusion medicine. We determined in vitro activity, specificity, and titer of anti-DEA 7 antibodies in DEA 7-negative dogs. Plasma samples from 317 DEA 7-negative dogs were cross-matched with DEA 7-positive red blood cells (RBCs) using gel column technology. Agglutination occurred with DEA 7-positive RBCs but not with DEA 7-negative RBCs in 73 samples (23%), which were hence classified as containing anti-DEA 7 antibodies. These samples were evaluated for hemolytic and agglutinating activity, strength of agglutination, and antibody specificity and titers. All samples showed agglutination but none showed hemolysis. Gel agglutination was graded as 1+ for 20 samples (27%), 2+ for 49 samples (67%), 3+ for 4 samples (6%); no samples were graded 4+. The agglutination titer was <1:2 for 51 samples (73%), 1:2 for 13 samples (19%), 1:4 for 4 samples (5%), and 1:8 for 2 samples (3%). Of 16 samples treated with 2-mercaptoethanol, 11 samples (69%) contained only IgM, 4 samples (25%) exhibited only IgG activity, and 1 sample (6%) had both IgG and IgM activity. Low titers of warm, weakly agglutinating, mostly naturally occurring IgM anti-DEA 7 antibodies were found in 23% of DEA 7-negative dogs. The presence of naturally occurring anti-DEA 7 antibodies suggests that cross-matching of canine blood recipients is advisable, even at first transfusion, to minimize delayed transfusion reactions., (© 2016 The Author(s).)

OBJECTIVE To determine the prevalence of naturally occurring anti-dog erythrocyte antigen (DEA) 7 antibodies in DEA 7-negative dogs from Spain and Italy. ANIMALS 252 DEA 7-negative dogs from a population of 312 dogs that were previously tested for DEA 1, DEA 4, and DEA 7. PROCEDURES A plasma sample was obtained from each dog and evaluated for anti-DEA 7 antibodies by the use of gel column agglutination. Each plasma sample underwent major crossmatching with RBCs from DEA 7-positive dogs. Samples that resulted in agglutination were then crossmatched with RBCs from DEA 1-negative, DEA 4-positive, and DEA 7-negative dogs to confirm the presence of anti-DEA 7 antibodies. Results were then used to calculate the risk for a delayed transfusion reaction in a DEA 7-negative dog with anti-DEA 7 antibodies after a transfusion with blood that was not crossmatched or typed for DEA 7. RESULTS 96 of 252 (38.1%) plasma samples contained anti-DEA 7 antibodies. A DEA 7-negative dog with anti-DEA 7 antibodies had a 5.9% chance of developing a delayed hemolytic reaction after transfusion with blood not crossmatched or typed for DEA 7. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that canine blood used for transfusion should be crossmatched with the blood or plasma of the intended recipient prior to transfusion to minimize the likelihood that the recipient will develop a hemolytic reaction associated with anti-DEA 7 antibodies. Ideal canine blood donors should be negative for both DEA 1 and DEA 7.

Aim: The aim of this study was to evaluate changes in proteinuria in dogs naturally infected with visceral leishmaniasis, following treatment with miltefosine (MLF) and allopurinol., Materials and Methods: Medical records of 40 dogs with leishmaniasis, treated with 2 mg/kg MLF every 24 h PO and 10 mg/kg allopurinol every 12 h for 28 days were reviewed. 20 dogs were included in the study, and clinical staging was performed following guidelines of the Canine leishmaniasis (CanL) Working Group, and dogs were categorized for proteinuria according to the International Renal Interest Society (IRIS) staging system. Clinical score, indirect fluorescent antibody test titer, serum total protein, gamma globulin (IgG), serum creatinine and urea concentration, and urine protein creatinine ratio (UP/C) were recorded at the time of diagnosis before the start of therapy (D0) and at the end of 28 days of therapy (D28)., Results: Following the CanL Working Group staging, all 20 dogs were classified as the clinical Stage C (Clinical disease) before and after the cycle of treatment. Before the cycle of therapy, dogs were categorized according to the IRIS staging system, as: 9/20 non-proteinuric (NP), 7/20 borderline proteinuric (BP), and 4/20 proteinuric (P). After treatment, 12/20 dogs were NP, 7/20 were BP, and 1/20 was P. There was a significant change in UP/C values before and after one cycle of treatment with MLF. In detail, after 28 days of therapy, 2 of 9 NP dogs became BP, 3 of the 7 BP dogs became NP, and 2 of the 4 P dogs became NP., Conclusion: This study showed a significant decrease in UP/C values occurred after one cycle of treatment with MLF and allopurinol in dogs naturally affected with CanL. This suggests that MLF does not increase proteinuria, and the use of MLF could be considered for the management of dogs with leishmaniasis, particularly in those with impaired renal function at the time of diagnosis.

The enzyme cytidine monophospho-N-acetylneuraminic acid hydroxylase is associated with the production of sialic acids on cat red blood cells. The cat has one major blood group with three serotypes; the most common blood type A being dominant to type B. A third rare blood type is known as AB and has an unclear mode of inheritance. Cat blood type antigens are defined, with N-glycolylneuraminic acid being associated with type A and N-acetylneuraminic acid with type B. Blood type AB is serologically characterized by agglutination using typing reagents directed against both A and B epitopes. While a genetic characterization of blood type B has been achieved, the rare type AB serotype remains genetically uncharacterized. A genome-wide association study in Ragdoll cats (22 cases and 15 controls) detected a significant association between blood type AB and SNPs on cat chromosome B2, with the most highly associated SNP being at position 4,487,432 near the candidate gene cytidine monophospho-N-acetylneuraminic acid hydroxylase. A novel variant, c.364C>T, was identified that is highly associated with blood type AB in Ragdoll cats and, to a lesser degree, with type AB in random bred cats. The newly identified variant is probably linked with blood type AB in Ragdoll cats, and is associated with the expression of both antigens (N-glycolylneuraminic acid and N-acetylneuraminic acid) on the red blood cell membrane. Other variants, not identified by this work, are likely to be associated with blood type AB in other breeds of cat.

Stray cats in the city of Milan, Italy, were tested for Leishmania infantum and other selected infections. Twenty-seven cats (30.0%) were seroreactive by indirect fluorescent antibody test (IFAT), with an antibody titer of 1:40 for 16 (17.7%) cats and 1:80 (cut-off for feline L. infantum infection) for 11 (12.2%) cats. One blood (1.1%) and one popliteal lymph node (1.1%) sample tested positive by real-time polymerase chain reaction; no oculoconjunctival swabs tested positive. Feline immunodeficiency virus, feline leukemia virus, and feline coronavirus (FCoV) seroprevalence determined by enzyme-linked immunosorbent assay was 6.1, 6.1, and 39.0%, respectively. Toxoplasma gondii, Bartonella henselae, and Chlamydophila felis prevalence determined by IFAT was 29.3, 17.1, and 17.1%, respectively. The frequency of seroreactivity to L. infantum was significantly higher in FCoV-seropositive cats (OR=4.4, P=0.04). L. infantum-infected stray cats in Milan have a high seropositivity rate, comparable to that of cats in areas endemic for leishmaniosis., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Serum hyperviscosity syndrome (HVS) was documented in two dogs with canine leishmaniasis (CanL) and seizures as the major clinical complaint. In both cases, laboratory abnormalities included mild non-regenerative anemia, thrombocytopenia, hypoalbuminemia, hyperproteinemia with monoclonal gammopathy, and marked serum hyperviscosity. CanL was diagnosed using cytology in one case and indirect immunofluorescence assay and conventional polymerase chain reaction in the second. Specific therapy with meglumine antimoniate and allopurinol(c) led to short-term remission in both dogs and normalization of serum viscosity. Although dogs rarely develop HVS, it should be suspected if hyperproteinemia and monoclonal gammopathy are present. Since CanL manifests with a variety of clinical presentations, including seizures resulting from HVS-induced central nervous system hypoxia, it should also be considered as a differential diagnosis in animals with seizures as a primary presenting sign.

The aims of this study were to evaluate the prevalence of Dog Erythrocyte Antigens (DEA) 1, 4, and 7 in Ibizan hounds, to compare the results with the prevalence of DEA in Spanish greyhounds, and to determine the risk of sensitization following the first transfusion of blood not typed for DEA 1 and the probability of an acute hemolytic reaction following a second incompatible transfusion using untyped DEA 1 blood. DEA 1, 4, and 7 status was determined in 92 Ibizan hounds. Results were compared with the previously reported prevalence in Spanish greyhounds. The risks of sensitization and of a hemolytic transfusion reaction were determined amongst Ibizan hounds and between Ibizan hounds and Spanish greyhounds. The prevalence of DEA 1, 4, and 7 was 75%, 98.9%, and 25%, respectively. There was a significantly higher expression of DEA 1 and 7 in Ibizan hounds than in Spanish greyhounds. The probability of sensitization of a recipient dog to DEA 1 with transfusions amongst Ibizan hounds was 18.5% and between Ibizan hounds and Spanish greyhounds was 13.7%. The probability of an acute hemolytic reaction in each group was 3.5% and 1.9%, respectively. There is a higher prevalence of DEA 1 and 7 in Ibizan hounds than in other sighthounds.

Objective: To determine the accuracy of an immunochromatographic cartridge (IC) test for blood typing feline type A, B, and AB blood samples., Design: Prospective observational study., Setting: University teaching hospital., Samples: Fifty-one nonanemic and 19 anemic feline blood samples., Interventions: Samples were blood typed by both card agglutination (CA) and IC techniques. Discordant results were analyzed using a back-typing technique for the presence of alloantibodies. Repeatability and reproducibility of the IC method were evaluated. Accuracy of the IC method was determined for feline whole blood anticoagulated with EDTA and citrate phosphate dextrose adenine (CPDA1), for feline-packed RBCs with CPDA1 and saline adenine glucose-mannitol, and for autoagglutinated and hemolytic samples. Accuracy of IC testing was determined for feline blood after room temperature, 4 ± 2, -20, and -80°C storage following 24, 48, 72, and 96 hours, and after 1, 2, 3, and 4 weeks of storage., Measurements and Main Results: In anemic and nonanemic samples the IC technique had a specificity, sensitivity, positive predictive value, and negative predictive value of 100% in detecting feline blood types A, B, and AB, outperforming a CA test. Results were repeatable and reproducible. Using IC it was possible to blood type samples anticoagulated with EDTA and CPDA1, packed RBC samples with CPDA1 and saline adenine glucose-mannitol, autoagglutinated and hemolytic samples, and samples stored at 4 ± 2°C and at room temperature for up to 1 month., Conclusions: The IC technique is an accurate assay for the identification of A, B, and AB blood types in anemic and nonanemic feline blood. It has a higher sensitivity and specificity than the CA test, and can be used in samples stored with common anticoagulants or preservative solutions used in feline transfusion medicine., (© Veterinary Emergency and Critical Care Society 2015.)

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., (© ISFM and AAFP 2015.)

Galgos (Spanish Greyhounds), in common with other sighthounds, have higher hematocrits, hemoglobin concentrations, and red blood cell counts than other breeds. In addition to these hematological characteristics, the physical characteristics of these dogs (medium to large dogs with an easily accessible jugular vein and a good temperament) make galgos ideal blood donors. However, to date, there are only published reports concerning dog erythrocyte antigen (DEA) 1 in this breed. Information on DEAs 4 and 7 would be useful when recruiting blood donors to donation programs, as DEA 1 and 7-negative and DEA 4-positive dogs can be considered universal donors. Ethylenediamine tetra-acetic acid-anticoagulated jugular blood samples were collected from 205 galgos. Dogs were aged between 1 and 10 years, 102 were female (49.8%) and 103 male (50.2%), and all were living in South Madrid, Spain. All 205 blood samples were tested for DEA 1 by card agglutination, and 150 of these samples were tested for DEA 4 and DEA 7 by gel column agglutination using polyclonal anti-DEA antibodies. Of the 205 galgos blood samples typed, 112 out of 205 (54.6%) were positive for DEA 1. Of the 150 blood samples tested, all (150/150, 100%) were positive for DEA 4, and 12 out of 150 (8%) samples tested positive for DEA 7. Of these samples, 70 out of 150 (46.7%) were positive only for DEA 4. There was no relationship between blood types and sex. In addition to the hematological characteristics previously reported and the physical characteristics of these dogs, the relative prevalence of blood types DEA 1, 4, and 7 make galgos good candidates for blood donation in blood donor programs., (© 2015 The Author(s).)

This prospective study investigated the effect on clinical and haematological variables of the anaesthetic combination of tiletamine and zolazepam in feline blood donors. Blood (10 ml/kg bodyweight to a maximum volume of 60 ml) was collected from the jugular vein of 31 owned healthy cats anaesthetised with 2.5 mg/kg of tiletamine and 2.5 mg/kg of zolazepam intramuscularly. Rectal temperature (RT), systolic arterial pressure (SAP), mean arterial pressure (MAP), diastolic arterial pressure (DAP), heart rate (HR) and complete blood count (including red blood cells [RBC], haemoglobin [HB], haematocrit [HT], platelet [PLT] count, white blood cells [WBC], lymphocytes, neutrophils, eosinophils, monocytes and basophils) were evaluated pre- and postdonation. RT decreased significantly (P <0.01) after blood donation (mean change in RT -0.7°C). Significant increases in SAP (P = 0.03), MAP (P <0.01) and DAP (P <0.01) occurred after blood donation (mean increase 13 mmHg, 12 mmHg and 11 mmHg, respectively). Although RBC, HT, HB, WBC, PLT, neutrophil and monocyte counts decreased, and HR, and lymphocyte, eosinophil and basophil counts increased after blood donation this change was not statistically significant. Mean time from pre- to postdonation evaluation was 39 ± 11 mins (range 24-76 mins). None of the cats had evidence of pallor or collapse after recovery from anaesthesia. The collection of blood at 10 ml/kg bodyweight to a maximum volume of 60 ml in healthy cats using a low dose tiletamine and zolazepam anaesthetic appears to be well tolerated by feline blood donors., (© ISFM and AAFP 2014.)

Feline vector-borne diseases can be caused by a range of pathogens transmitted by arthropods. Many of these infections have zoonotic implications, and stray cats are potential sentinels for human and pet health. This study investigated the prevalence of selected vector-borne infections in stray colony cats in Milan. Blood samples from 260 stray cats were evaluated, using conventional polymerase chain reaction tests (cPCRs), for the presence of DNA associated with Rickettsia species, Anaplasma phagocytophilum and Ehrlichia species. Positive cPCR results occurred in 127/260 subjects (48.9%; 95% confidence interval [CI] = 40.7-58.1), with a prevalence of 31.9% (83/260, 95% CI = 25.4-39.6) for Rickettsia species, 17.7% (46/260, 95% CI= 13.0-23.6) for A phagocytophilum, and 5.4% (14/260, 95% CI = 2.9-9.0) for Ehrlichia species. There was no statistical association between a positive PCR test for vector-borne infections surveyed and colony location, age, gender, body condition score or complete blood count abnormalities, nor feline immunodeficiency virus, feline leukaemia virus or Toxoplasma gondii status. The only variable linked to positive PCR results was detection of signs of ocular infection and PCR positivity for Rickettsia species (P = 0.04, odds ratio [OR] = 2.2, 95% CI = 1.1-4.4, P = 0.02). There is a significant prevalence of vector-borne infections with zoonotic potential in urban stray cats in Milan. Thus, dogs and pet cats with outdoor access should be monitored and treated for ectoparasites on a regular basis to minimise risks of disease and the potential transmission of zoonotic agents to people., (© ISFM and AAFP 2013.)

Feline otitis externa is a dermatological disorder that has not been evaluated much in stray cats. One hundred and eighty-seven stray cats were randomly selected during a trap-neuter-release programme to investigate the prevalence of otitis externa in stray cat colonies in northern Italy. Swabs for cytological examination were obtained from the external ear canal of each cat. A direct otoscopic assessment of the external ear canal was made in 86/187 cats. Cytological evidence of otitis externa was present in 55.1% of cats. The influence on otitis of age, gender, habitat and season of sampling was tested, but no risk factors were found. Otodectes cynotis (as a sole agent or in combination) was the primary cause of otitis in 53.3% of cats. Cocci and rods, either alone or in combination with other agents, were perpetuating factors in 71.8% and 29.1% of cats, respectively. Pregnancy status was a risk factor for otitis caused by coccal infections. Malassezia species, alone or in combination, was the perpetuating factor in 50.5% of cats with otitis. Urban habitat and winter season were risk factors for otitis associated with Malassezia species. Demodex cati was identified as an incidental finding in two cats. There was good agreement between otoscopy and cytology with regard to the diagnosis of otitis externa. The results of this study show a high prevalence of otitis externa in stray colony cats and provide information on causal factors for feline otitis externa., (© ISFM and AAFP 2013.)

Background: Few studies have investigated the effect of veterinary clinical procedures on the welfare of dogs, with specific emphasis on the veterinary practice environment. Clinicopathologic variables have also not been assessed in these potentially stressful situations. Similar to human clinical studies, the veterinary clinical waiting room could present a significant stress factor for dogs., Objectives: The present study was designed to investigate the effect of waiting-room environment on serum cortisol and glucose alterations as well as heart rate in privately owned healthy dogs., Methods: The clinical trial included 24 healthy dogs that were divided into 2 groups: the clinical waiting-room group (A) and the control group (B) that waited outside in a garden. During the entire experiment, 18 dogs (9 dogs per group) were monitored with a human heart rate monitor fastened around the chest. After 20 minutes of waiting, blood samples were collected from all of the dogs (24 dogs) to determine serum cortisol concentration., Results: Serum cortisol concentration and mean, maximum, and minimum heart rate were significantly higher in group A compared with group B, but there was no statistical difference in serum glucose concentrations between the 2 study groups., Conclusions: Results of this study suggest that the waiting room is a potentially stressful situation for dogs in clinical veterinary practice, when compared with a garden, based on the assessment of adrenal cortex function and heart rate evaluation., (© 2014 American Society for Veterinary Clinical Pathology and European Society for Veterinary Clinical Pathology.)

This study investigated the prevalence of feline haemoplasma infections in a number of stray cat colonies in Milan, Northern Italy. Blood samples from 260 stray cats were evaluated, with conventional PCR, for the presence of DNA associated with Mycoplasma haemofelis (Mhf) and "Candidatus Mycoplasma haemominutum" (CMhm). Odd ratios (OR) were calculated to identify risk factors for haemoplasma infections. PCR was positive in 86 out of 260 subjects (33.1%), with a prevalence of 10.8% (28/260 cats) for Mhf and 22.3% (58/260 cats) for CMhm. No coinfections were registered. There were significant associations between infections and season of sampling, that is, a negative association between winter sampling and a haemoplasma positive status (OR = 0.29, P = 0.001), or CMhm positive status (OR = 0.29, P = 0.01). Haemoplasma infections are common in stray cats in Milan. Thus, domestic cats with outdoor access should be routinely monitored and treated for ectoparasites to minimize risks of disease acquisition. Moreover, as these infections are transmitted via blood, feline blood donors from this area should be screened by PCR and preferably be drawn from a population of indoor cats regularly treated for fleas.

Emerging diseases caused by piroplasms pose a health risk for man and other animals, and domestic cats have been proposed as potential reservoirs for some piroplasm infections. The aim of this study was to identify the frequency of the piroplasms Babesia microti and Cytauxzoon felis in stray cats from northern Italy and to identify possible risk factors associated with these infections. Blood samples from 260 stray cats enrolled in a trap-neuter-release (TNR) program in northern Italy were examined with conventional PCR for the presence of Babesia microti and Cytauxzoon felis DNA. No sample (0.0%) tested positive for C. felis, whilst B. microti DNA was detected in two samples (0.8%). Both infected cats were in good clinical condition and recovered well from the neutering surgery. One of these two cats had a triple coinfection with Babesia microti, Candidatus Mycoplasma haemominutum, and Anaplasma phagocytophilum. Evidence presented in this study indicates that the blood borne protozoans Babesia microti and Cytauxzoon felis are not widely distributed in stray cat populations in Milan, northern Italy, and that the significance of cats as a reservoir host for B. microti in this area is limited.

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.