European multicenter study on antimicrobial resistance in bacteria isolated from companion animal urinary tract infections.

BACKGROUND: There is a growing concern regarding the increase of antimicrobial resistant bacteria in companion animals. Yet, there are no studies comparing the resistance levels of these organisms in European countries. The aim of this study was to investigate geographical and temporal trends of antimicrobial resistant bacteria causing urinary tract infection (UTI) in companion animals in Europe. The antimicrobial susceptibility of 22 256 bacteria isolated from dogs and cats with UTI was determined. Samples were collected between 2008 and 2013 from 16 laboratories of 14 European countries. The prevalence of antimicrobial resistance of the most common bacteria was determined for each country individually in the years 2012-2013 and temporal trends of bacteria resistance were established by logistic regression. RESULTS: The aetiology of uropathogenic bacteria differed between dogs and cats. For all bacterial species, Southern countries generally presented higher levels of antimicrobial resistance compared to Northern countries. Multidrug-resistant Escherichia coli were found to be more prevalent in Southern countries. During the study period, the level of fluoroquinolone-resistant E. coli isolated in Belgium, Denmark, France and the Netherlands decreased significantly. A temporal increase in resistance to amoxicillin-clavulanate and gentamicin was observed among E. coli isolates from the Netherlands and Switzerland, respectively. Other country-specific temporal increases were observed for fluoroquinolone-resistant Proteus spp. isolated from companion animals from Belgium. CONCLUSIONS: This work brings new insights into the current status of antimicrobial resistance in bacteria isolated from companion animals with UTI in Europe and reinforces the need for strategies aiming to reduce resistance.

Lymphocytic leukemia in a captive dhole (Cuon alpinus).

A captive 3-yr-old male dhole (Cuon alpinus) was presented for poor body condition. Pancytopenia concurrent with bone marrow aspiration that revealed severe medullary infiltration by a population of initially small lymphocytes was diagnostic of an aleukemic chronic lymphocytic leukemia. Chemotherapy was initiated, but euthanasia was elected after the animal's rapid deteriorating condition and sudden lymphoid organs hypertrophy several days after initial presentation. Histology revealed lymphoid organs and bone marrow infiltration by highly proliferating immature lymphocytes compatible with a blast crisis. On immunohistochemistry, neoplastic cells appeared CD3 positive, confirming a T lymphoid origin. This is the first report of a lymphocytic leukemia in a wild canid species.

Detection of SARS-CoV-2 in two cats during the second wave of the COVID-19 pandemic in France.

Although there are several reports in the literature of SARS-CoV-2 infection in cats, few SARS-CoV-2 sequences from infected cats have been published. In this study, SARS-CoV-2 infection was evaluated in two cats by clinical observation, molecular biology (qPCR and NGS), and serology (microsphere immunoassay and seroneutralization). Following the observation of symptomatic SARS-CoV-2 infection in two cats, infection status was confirmed by RT-qPCR and, in one cat, serological analysis for antibodies against N-protein and S-protein, as well as neutralizing antibodies. Comparative analysis of five SARS-CoV-2 sequence fragments obtained from one of the cats showed that this infection was not with one of the three recently emerged variants of SARS-CoV-2. This study provides additional information on the clinical, molecular, and serological aspects of SARS-CoV-2 infection in cats.

First Evidence of Natural SARS-CoV-2 Infection in Domestic Rabbits.

We tested 144 pet rabbits sampled in France between November 2020 and June 2021 for antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by microsphere immunoassay. We reported the first evidence of a natural SARS-CoV-2 infection in rabbits with a low observed seroprevalence between 0.7% and 1.4%.

Comparison of STA-NeoPTimal (Stago) and STA-Neoplastine CI Plus (Stago) thromboplastin reagents using a STA Satellite Max analyzer to measure prothrombin times in dogs.

BACKGROUND: Different thromboplastins are available to measure prothrombin time (PT). Stago coagulation analyzers and reagents are currently used in veterinary laboratories and enable PT measurements to explore the coagulation cascade (extrinsic pathway). OBJECTIVES: The main objective was to compare PT measurements obtained with the STA-NeoPTimal reagent with the commonly used STA-Neoplastine CI Plus reagent. The secondary objective was to compare the PT ratio with the international normalized ratio (INR) calculated from our derived clotting times. METHODS: Analytical performance was evaluated with intra-assay and inter-assay precision. Seventy-two individual canine plasma samples were collected. Each sample was tested with both thromboplastins, using an STA Satellite Max analyzer. The PT, PT ratio, and INR values obtained with the two reagents were compared using Passing-Bablok regression for correlations and Bland-Altman plots for method agreements. RESULTS: The analytical performance of STA-NeoPTimal reagent was acceptable. Compared with the STA-Neoplastine CI Plus reagent, the STA-NeoPTimal reagent showed a positive proportional bias for PT values. Narrow range analyses showed good agreement for normal PT values (less than 9.5 seconds, internal reference cutoff with STA-Neoplastine CI Plus), and clinical concordance was achieved. When PT was prolonged (more than 9.5 seconds), PT increases were more marked with the STA-NeoPTimal reagent. Agreement was good for INR values across the whole range of PT results. CONCLUSION: STA-NeoPTimal can be reliably implemented in veterinary laboratories for canine PT measurements, as agreement between the PT results measured with the two reagents was clinically acceptable.

Is it possible to use expired tubes for routine biochemical analysis in dogs?

BACKGROUND: Expired collection tubes may be used inadvertently and resampling is not always possible. To date, studies have not been conducted in veterinary medicine to determine whether or not biochemical measurements obtained from specimens collected into expired tubes are accurate enough for clinical decision-making. OBJECTIVES: The aims of this preliminary study were to assess the impact of measuring routine plasma biochemical analytes in canine specimens collected in expired tubes and to investigate the relationship between post-expiration time and the magnitude of errors. METHODS: Blood specimens were collected from 61 dogs and aliquoted equally into tubes containing lithium heparin and gel. One tube was within the expiration date, and the other tube was up to 11 months post-expiration. Plasma was separated within 1 hour of specimen collection, and concentrations of urea, creatinine, total protein, albumin, total bilirubin, cholesterol, triglycerides, magnesium, calcium, phosphates, sodium, potassium, chloride, total CO(2), and fructosamine and activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase (ALP), γ-glutamyltransferase, lactate dehydrogenase, creatine kinase, amylase, and lipase were analyzed immediately and results compared. RESULTS: For most analytes there was no significant difference between results from specimens collected in non-expired and expired tubes. For ALP and lipase activities and fructosamine and total CO(2) concentrations, significant differences were found, and results obtained for fructosamine and total CO(2) from specimens in expired tubes may have led to erroneous interpretations. The effect of time since expiration was constant over time. CONCLUSIONS: When specimens are processed within 1 hour of collection, results of routine biochemical measurements of blood collected in lithium heparin tubes remain clinically valid for up to 11 months after expiration of tubes for the majority of analytes, except for ALP, lipase, fructosamine, and total CO(2).

Anaplasmosis in two dogs in France and molecular and phylogenetic characterization of Anaplasma phagocytophilum.

Two dogs in France were diagnosed with Anaplasma phagocytophilum infection by real-time PCR. The most remarkable hematologic and biochemical findings were severe thrombocytopenia, mild neutrophilia, morulae in neutrophils, and increased serum concentration of the α2-globulin fraction detected by agarose gel electrophoresis of serum proteins. Using sequencing of the partial 16S rRNA and ankA genes, molecular characterization of the A. phagocytophilum strains showed that the organisms from both dogs were identical to the European strains isolated from horses and people. Based on phylogenetic analysis, the ankA gene was more discriminating than the 16S rRNA gene in distinguishing the majority of European and American strains of A. phagocytophilum infecting people and animals. Three isolates of A. phagocytophilum, 1 from Spain (cow) and 2 from Norway (sheep and deer), were external to the European and American clades.

Identification of JAK2 mutations in canine primary polycythemia.

OBJECTIVE: Primary polycythemia in dogs is classified as a myeloproliferative syndrome with a chronic progressive course and unspecific symptoms. Diagnosis is based on exclusion criteria. In humans, the presence of an acquired recurrent mutation within the JAK2 gene has recently been identified in 90% of the patients with polycythemia vera. This mutation (V617F) is located in the pseudokinase domain of JAK2, leading to constitutive activation of the kinase responsible for the polycythemia. Detection of the mutation has now become a major diagnostic tool in humans for polycythemia vera diagnosis. As the canine JAK2 gene shares strong homology with its human counterpart, we looked for the presence of JAK2 mutations in dogs with an elevated hematocrit. MATERIALS AND METHODS: Direct sequencing of the JAK2 exon 14 was performed on DNA extracted from the peripheral blood of five dogs suspected of primary polycythemia. Mutant subclones were expressed in interleukin-3-dependent BaF3 cells and tested for cytokine independency. RESULTS: One dog presented with a three-base change in codons 617 and 618 of JAK2 giving rise to V617F and C618L mutations. By polymerase chain reaction product subcloning, we demonstrated the coexistence of the wild-type sequence and a triple mutant sequence, while DNA from buccal swab contained the wild-type sequence only. Transfection of BaF3 cells with the triple mutant cDNA, but not with the wild-type complementary DNA, resulted in cytokine-independent growth and constitutive signal transducer and activation of transcription 5 phosphorylation. CONCLUSIONS: Identical mutations of the JAK2 gene occur in humans and dogs, giving rise to a constitutively active JAK2 kinase, suggesting a common mechanism for human and canine diseases. Thus, common diagnostic tools and therapeutic approaches may be relevant.