First description of reactive arthritis secondary to leptospirosis in a dog.

BACKGROUND: Leptospirosis is a zoonotic bacterial infection that is common worldwide, with a wide spectrum of clinical signs. It commonly infects the kidneys and the liver but can damage a number of organ systems. CASE DESCRIPTION: An 18-month-old boxer dog was referred because of reluctance to walk and sickness. FINDINGS/TREATMENT AND OUTCOME: His clinical presentation, including swollen and inflamed joints fulfilled the requirements for a diagnosis of immune-mediated polyarthritis (IMPA). Shortly after, unexpected icterus developed and laboratory signs of hepatic and renal failure were observed. A diagnosis of leptospirosis was reached after observing typical clinical signs, along with a positive microagglutination test. Since the diagnostic molecular test for Leptospira from joint fluids came back negative and also the localization of Leptospira in multiple joints in association with inflammation has never been described in canine patients, an immune-mediated complication seemed most likely. The dog quickly recovered after the administration of ampicillin for 5 days, followed by a two week course of doxycicline. CONCLUSION: In human medicine, this case would be considered as a reactive arthritis (ReA), which is mistakenly cited in the current veterinary literature for cases associated with chronic infections. In humans, ReA is described as inflammatory arthritis not directly caused by culture-proven infection of joint tissue but by infection at another site due to a complex interplay of host antimicrobial factors. This case presentation reports for the very first time, a case of canine leptospirosis mimicking an IMPA and fitting the description of ReA in human medicine.

Proposal for rational antibacterial use in the diagnosis and treatment of dogs with chronic diarrhoea.

Chronic diarrhoea is a frequent complaint in canine practice and the diagnostic path is often characterised by numerous diagnostic tests and stepwise empirical treatments, often applied before gastrointestinal endoscopy/mucosal biopsies. These include dietary interventions (novel protein, hydrolysed protein diet), parasiticides and still, in many cases, antibacterials. Indiscriminate use of antibacterial drugs risks detrimental consequences for both the individual patient (antimicrobial resistance, long-term disruption of intestinal bacterial populations, potential worsening of gastrointestinal signs) and the general public. For that reason, in this Perspective essay we advocate use of antibacterials only after histopathologic evaluation of gastrointestinal biopsies or, for those cases in which endoscopy is not possible, after other therapeutic trials, such as diet/pre-probiotics or anti-inflammatory drugs have proven unsuccessful. They should be reserved, after appropriate dietary trials, for those canine chronic diarrhoeic patients with signs of true primary infection (i.e. signs of systemic inflammatory response syndrome or evidence of adherent-invasive bacteria) that justify antibacterial use.

Use of acute phase proteins for the clinical assessment and management of canine leishmaniosis: general recommendations.

BACKGROUND: Dogs with canine leishmaniosis (CanL) due to Leishmania infantum can show a wide spectrum of clinical and clinicopathological findings at the time of diagnosis. The aim of this paper is to describe the possible application of acute phase proteins (APPs) for the characterization and management of this disease, based on previously published information on the utility of APPs in CanL and the experience of the authors in using APPs as analytes in the profiling of canine diseases. MAIN BODY: Dogs diagnosed with L. infantum infection by serology, polymerase chain reaction, cytological or histopathological identification, can be divided into three groups based on their clinical condition at physical examination and their APPs concentrations: Group 1: dogs with no clinical signs on physical examination and APPs in reference range; Group 2: dogs with changes in APPs but no clinical signs on physical examination; Group 3: dogs with clinical signs and changes in APPs. This report describes the main characteristics of each group as well as its association with the clinical classification schemes of CanL. CONCLUSION: APPs concentration can be a useful clinical tool to characterize and manage CanL.

Frequency of DEA 1 antigen in 1037 mongrel and PUREBREED dogs in ITALY.

BACKGROUND: The prevalence of dog erythrocyte antigen (DEA 1) in canine population is approximately 40-60%. Often data are limited to a small number of breeds and/or dogs. The aims of this study were to evaluate frequency of DEA 1 in a large population of purebred and mongrel dogs including Italian native breeds and to recognize a possible association between DEA 1 and breed, sex, and genetic and phenotypical/functional classifications of breeds. Frequencies of DEA 1 blood group collected from screened/enrolled blood donors and from healthy and sick dogs were retrospectively evaluated. The breed and the sex were recorded when available. DEA 1 blood typing was assessed by immunocromatographic test on K3EDTA blood samples. The prevalence of DEA 1 antigen was statistically related to breed, gender, Fédération Cynologique Internationale (FCI) and genotypic grouping. RESULTS: Sixty-two per cent dogs resulted DEA 1+ and 38% DEA 1-. DEA 1- was statistically associated with Dogo Argentino, Dobermann, German Shepherd, Boxer, Corso dogs, the molossian dogs, the FCI group 1, 2 and 3 and the genetic groups "working dogs" and "mastiff". DEA 1+ was statistically associated with Rottweiler, Briquet Griffon Vendéen, Bernese mountain dog, Golden Retriever, the hunting breeds, the FCI group 4, 6, 7 and 8 and the genetic groups "scent hounds" and "retrievers". No gender association was observed. CONCLUSIONS: Data obtained by this work may be clinically useful to drive blood donor enrollment and selection among different breeds.

Urinary excretion of calcium and phosphate in dogs with pituitary-dependent hypercortisolism: case control study in 499 dogs.

Pituitary-dependent hypercortisolism (PDH) in dogs is frequently associated with high serum phosphate and parathormone concentrations which are in turn associated with prognosis and clinical presentation. The pathogenesis of such abnormalities remains unknown. The aim of the present study was to evaluate the serum and urinary concentrations and the urinary fractional excretion of phosphate and calcium in dogs with PDH. Medical records of newly diagnosed PDH dogs before treatment from one referral centre were retrospectively evaluated. One clinically normal and one sick dog for each dog with PDH were included as controls. One hundred and sixty-seven dogs with PDH were included. The serum phosphate concentration in PDH dogs was significantly (P<0.0001) higher compared with clinically normal control dogs (CNDs) and sick control dogs (SCDs). The serum calcium concentration in PDH dogs was significantly higher compared with SCDs but not different compared with CNDs. Urinary fractional excretion of phosphate in PDH dogs was significantly lower compared with CNDs and SCDs. Urinary fractional excretion of calcium in PDH dogs was significantly higher compared with CNDs and SCDs. In conclusion, PDH dogs have lower phosphaturia and higher calciuria compared with control dogs. These findings suggest that, at least in part, high serum phosphate concentrations are related to the renal retention of phosphate.

An open-label phase 1 dose-escalation clinical trial of a single intravenous administration of gemcitabine in dogs with advanced solid tumors.

BACKGROUND: A broad range of gemcitabine dosages have been used in dogs. HYPOTHESIS/OBJECTIVES: To determine maximally tolerated dose (MTD), dose-limiting toxicity (DLT), and preliminary antitumor activity of intravenous administration of gemcitabine in dogs with advanced solid tumors. ANIMALS: Twenty-two client-owned dogs. METHODS: Dogs with advanced cancer were prospectively enrolled in an open-label Phase 1 study of gemcitabine. Gemcitabine was administered as a 30-minute intravenous bolus starting at 800 mg/m(2), using escalation of 50 mg/m(2) increments with 3 dogs per dose level. MTD was established based on the number of dogs experiencing DLT assessed after 1 cycle. Treatment continued until disease progression or unacceptable toxicosis. Additional dogs were enrolled at MTD to better characterize tolerability, and to assess the extent and duration of gemcitabine excretion. RESULTS: Twenty-two dogs were treated at 4 dose levels, ranging from 800 to 950 mg/m(2). Neutropenia was identified as DLT. MTD was 900 mg/m(2). DLT consisting of grade 4 febrile neutropenia was observed at 950 mg/m(2) in 2 dogs. There were no nonhematologic DLTs. Twenty dogs received multiple doses, and none had evidence of severe toxicosis from any of their subsequent treatments. At 900 mg/m(2), 2 complete and 5 partial responses were observed in dogs with measurable tumors. The amount of gemcitabine excreted in urine decreased over time, and was undetectable after the first 24 hours. CONCLUSIONS AND CLINICAL IMPORTANCE: The recommended dose of gemcitabine for future Phase 2 studies is weekly 900 mg/m(2). In chemotherapy-naïve dogs with advanced solid tumor this dose level merits further evaluation.

Spontaneous acromegaly: a retrospective case control study in German shepherd dogs.

Acromegaly results from the overproduction of growth hormone in adulthood and is characterised by overgrowth of soft tissue and/or bone as well as insulin resistance. There are few data indicating the risk factors associated with this disease in dogs or its clinicopathological features and sequelae. The objective of this retrospective study was to catalogue and assess these aspects of the disease in German shepherd dogs (GSDs) which were found to be over-represented among acromegalic dogs attending two veterinary referral clinics over a period of 7 years. Each acromegalic dog (AD) was compared with two breed/age/sex matched controls. Clinical signs of acromegaly included panting, polyuria/polydipsia, widened interdental spaces, weakness, inspiratory stridor, macroglossia, weight gain, redundant skin folds, thick coat, exophthalmos and mammary masses. Serum alkaline phosphatase, creatine-kinase, glucose, triglyceride, phosphate ion, and 'calcium per phosphate product' concentrations were significantly higher in acromegalic animals while haemoglobin concentration, blood urea nitrogen, sodium and chloride ion concentrations, and urinary specific gravity, osmolality and fractional excretion of phosphate were significantly lower. Although, in the majority of cases clinicopathological abnormalities resolved following ovariohysterectomy, in one dog, acromegalic signs abated and insulin-like growth factor-1 concentrations normalised only following the surgical excision of mammary tumours carried out 2 months after ovariohysterectomy. The findings of this study indicate that GSDs are predisposed to the development of acromegaly with a suspected inherited susceptibility.

Cytauxzoon sp. infection in the first endemic focus described in domestic cats in Europe.

Information about epidemiological and clinicopathological aspects of domestic cat infection by species of Cytauxzoon other than Cytauxzoon felis is limited and it has rarely been reported. Following the detection of clinical cytauxzoonosis in three cats from Trieste (Italy), an epidemiological study was carried out in colony (n=63) and owned (n=52) cats from the same city to investigate the presence of Cytauxzoon sp. infection and to assess clinicopathological findings and variables associated with this infection. Cytauxzoon sp. infection was detected by 18S rRNA gene PCR in 23% (27/118) and by blood smear examination in 15% (18/118) of domestic cats. The 18S rRNA gene sequences obtained were 99% identical to the Cytauxzoon sp. sequences deposited in GenBank(®) from Spanish, French and Mongolian wild and domestic cats. Erythroparasitemia was observed mainly in apparently healthy cats. Cytauxzoon sp. infection was statistically associated with the colony group and the outdoor life style. No statistical association was found between positivity by PCR and breed, gender, age, presence of ticks and/or fleas, clinical status, laboratory findings such as anemia, FIV and/or FeLV status and mortality rate. Persistence of the infection was monitored and documented in four clinical cases. We reported the first clinicopathological description of naturally occurring Cytauxzoon sp. infection in domestic cats living in Italy. The predominance of subclinical erythroparasitemia and the evidence of persistent infection support the hypothesis that the domestic cat might serve as a reservoir host for this infection.

Clinicopathological findings, molecular detection and characterization of Babesia gibsoni infection in a sick dog from Italy.

A 4-year-old intact female American Pit Bull Terrier from Italy descendant of an American-born bitch was evaluated for anorexia, lethargy, weakness, and intermittent vomiting. On physical examination, the dog was dehydrated, had pale mucous membranes, hunched posture and abdominal pain. A moderate anemia was observed. Splenomegaly and hyperechoic regions suspected as infarcts in the spleen were seen on abdominal ultrasound. Based on the suspicion of splenic torsion, splenectomy was performed. After surgery, the clinical condition deteriorated. A follow-up complete blood count demonstrated severe macrocytic normochromic anemia with evidence of marked regeneration, left shift neutrophilia, monocytosis and marked thrombocytopenia. Blood smear evaluation revealed single to multiple, variable sized (1-3 microm in diameter), and round to oval to band-like piroplasms within many red blood cells consistent with small form Babesia spp. or Theileria spp. A partial segment of the 18S rRNA gene was amplified and the PCR product was analyzed by direct sequencing. The nucleotide sequence was completely identical to that of Babesia gibsoni present in GenBank. This is the first molecular detection and characterization of B. gibsoni infection in a sick dog from Italy.

Detection of erythrocyte binding IgM and IgG by flow cytometry in sick dogs with Babesia canis canis or Babesia canis vogeli infection.

The aim of this study was to examine by means of flow cytometry immunophenotyping (FCI) if sick dogs infected with Babesia canis canis (B. c. canis) or Babesia canis vogeli (B. c. vogeli) had anti-erythrocyte membrane binding IgG and/or IgM at the time of diagnosis. Diagnosis of Babesia infection was assessed by blood smear and by PCR-restriction fragment length polymorphism analysis in 30 sick dogs. Signalment, clinical history, physical examination and laboratory tests of B. c. canis (n=24) and of B. c. vogeli (n=6) infected dogs were studied. The majority of B. c. canis infected dogs showed anemia (92%) predominantly non-regenerative (94%), while the B. c. vogeli infected dogs had a regenerative anemia (67%). Eccentrocytosis was present in 33% of the B. c. canis infections. Four of six B. c. vogeli infected dogs had erythrocytes membrane antibodies. One dog resulted uncertain and one resulted negative to FCI. In contrast, all the B. c. canis infected dogs were negative for erythrocytes membrane binding immunoglobulins detection. In addition, the mean percentages of erythrocytes binding IgG and IgM were statistically much lower in B. c. canis than in B. c. vogeli infected dogs. At the time of the diagnosis, the formation of erythrocyte membrane binding IgG and IgM by immune mechanisms appears not to be involved in B. c. canis infections while it is present in the majority of B. c. vogeli infections.

Babesia canis canis and Babesia canis vogeli clinicopathological findings and DNA detection by means of PCR-RFLP in blood from Italian dogs suspected of tick-borne disease.

The aims of this study were to determine the presence of Babesia spp. in blood samples from Italian dogs with clinical signs compatible with tick-borne diseases by means of PCR-restriction fragment length polymorphism (RFLP) and describe the clinicopathological findings of dogs with Babesia infection. We evaluated the majority of canine babesiosis cases by means of clinical history, physical examination, hematological, biochemical, serum electrophoresis, urinalysis and hemostatic tests. Forty-five out of 164 canine blood samples studied were positive to Babesia PCR-RFLP with the following results: Babesia canis canis (n=34) and Babesia canis vogeli (n=11). The majority of B. c. canis infections were detected in Northern Italy (29.1%; 30/103). B. c. vogeli cases were detected mainly in Central and Southern Italy (16.3%; 10/61). Only one B. c. vogeli was detected in Northern Italy (0.9%; 1/103). Three positive samples to B. c. canis and four positive samples to B. c. vogeli were selected for sequencing of a fragment of the 18S rRNA gene (410bp) for further molecular characterization. The sequence obtained from all seven dogs was 99/100% homologous to sequences from B. c. canis and B. c. vogeli, respectively, present in GenBank. Sixty-two percent of dogs infected with B. c. canis had recently travelled on a hunting trip to East European countries. The main acute clinical signs were dehydration, apathy, anorexia and fever. The majority of dogs infected with B. c. canis presented at initial clinical examination mild to severe thrombocytopenia, hyperfibrinogenemia, mild to moderate normocytic-normochromic non-regenerative anemia, hemolysis and neutropenia. The urinalysis showed hemoglobinuria in 13/19 dogs suggesting intravascular hemolysis. Dogs with B. c. canis infection had high levels of C-reactive protein. Hypoalbuminemia was present in 17/26 dogs. The 11 cases of B. c. vogeli infection did not present a homogenous clinicopathological pattern. B. c. vogeli infections were observed in young dogs causing hemolytic anemia and in adult/old does that frequently presented predisposing factors such as splenectomy or immunocompromised conditions. In conclusion, this study demonstrates the presence of B. c. canis and B. c. vogeli in Italian sick dogs and differences in clinicopathological pattern in these two species of B. canis.

Molecular survey of Rickettsia spp. in sick dogs in Italy.

The aim of this retrospective study was to investigate the prevalence of Rickettsia spp. DNA in the blood of sick dogs from Italy. Canine blood samples (n=650) submitted for molecular testing of Rickettsia spp. to a diagnostic laboratory from February 2003 to March 2006 were studied. The Rickettsia spp. DNA detection was performed by Light Cycler real-time PCR using hybridization probes separately conducted with specific primers and probes. The total percentage of Rickettsia spp.-positive dog samples was 1.5% (10 out of 650). The percentage of Rickettsia spp.-positive dog samples submitted from north, central and southern Italy were 0.4% (1/248), 1.4% (3/219) and 3.3% (6/183), respectively. Five out of 138 dogs (3.6%) from Sicily were positive on Rickettsia PCR testing. A statistical difference was found between the percentages of positive samples from the Yorkshire terrier group (10.7%) compared with the mixed breed group (0.7%). No statistical differences were found between seasonal period, region and gender. Based on molecular data, there is infrequent rickettsiemia in dogs.

Molecular survey of Ehrlichia canis and Anaplasma phagocytophilum from blood of dogs in Italy.

We investigated the prevalence of Ehrlichia canis (E. canis) (n = 601) and Anaplasma phagocytophilum (A. phagocytophilum) (n = 460) infection by means of real-time PCR from blood of Italian dogs. The prevalence of E. canis in northern, central, and southern Italy was 2.9%, 8%, and 9.7%, respectively. The prevalence of A. phagocytophilum was 0%.

Nasal melanosis in three dogs.

Cytologically and histologically confirmed nasal melanosis was detected by rostrocaudal rhinoscopic evaluation of three dogs with unilateral nasal discharge caused by a chronic and severe odontopathic rhinitis. The extraction of affected teeth and prolonged antibiotic therapy led to a complete resolution of nasal disease. The nasal melanosis could be considered a partial metaplastic transformation of mucosal respiratory cells with accumulation of intracytoplasmic melanin.

Validation of an automated spectrophotometric assay for the determination of cholinesterase activity in canine serum.

The determination of enzymatic activity of cholinesterase is a useful diagnostic method to detect exposure to anticholinesterase compounds in human and in veterinary medicine. We validated a modification of the Ellman method in canine serum and applied it to the diagnosis of dogs poisoned with anticholinesterase substances. The method used butyrylthiocholine as substrate and potassium hexacyanoferrate as chromophore. The reference range calculated on 60 clinically healthy dogs was set between 3405 and 6561 U/L (chi-square test for normal distribution, p > 0.05). The overall mean intra-assay and inter-assay coefficients of variation were 0.53% and 3.83%, respectively. The assay was linear when using two sera with 12,538 U/L and 6604 U/L serum cholinesterase activity (r(2) = 0.997) and 0.999, respectively). The mean recovery values of pooled sera with a mean pseudocholinesterase (PChE) activity of 12,081 U/L and pooled sera with a mean PChE activity of 3415 U/L were 103.5% and 102.8%, respectively. Six dogs with a diagnosis of anticholinesterase compound intoxication showed a decrease in cholinesterase activity of at least 50% of normal activity with a mean +/- SD of 487 +/- 291 U/L ranging from 169 to 847 U/L. This technique conforms to the current standard for precision, linearity and accuracy and is a useful method for the complementary diagnosis of organophosphate or carbamate insecticide intoxication in dogs.

Diagnosis of canine nasal aspergillosis by cytological examination: a comparison of four different collection techniques.

OBJECTIVES: To compare the efficacy and diagnostic value of four different sample collection techniques for cytological identification of nasal aspergillosis-penicilliosis in dogs. METHODS: Fifteen dogs with a history of persistent nasal discharge and clinical and radiographic findings suggestive of aspergillosis were evaluated using four different cytological sampling techniques. These were a direct smear from the nasal discharge, blind swab collection under general anaesthesia, brushing from suspect lesions under direct endoscopic visualisation and a squash technique of mucosal biopsies from suspect lesions obtained under direct endoscopic visualisation. RESULTS: Direct smear collection and blind swab collection detected fungal hyphae in 13.3 and 20 per cent of examined cases, respectively; brush samples detected fungal hyphae in 93.3 per cent and fungal spores in the 45 per cent of examined cases and squash samples detected fungal hyphae in 100 per cent and fungal spores in 36 per cent of examined cases. CLINICAL SIGNIFICANCE: This study confirmed the high accuracy of cytology samples in the diagnosis of nasal aspergillosis-penicilliosis when collected under direct endoscopic visualisation and showed the poor value of samples that were collected by blind swabs or prepared from samples of nasal discharge.

Metallothionein expression in benign and malignant canine mammary gland tumours.

The presence of metallothioneins (MTs) were demonstrated immunohistochemically using a monoclonal antibody (E9) against a conserved epitope of I and II isoforms in canine mammary tumours. In a semiquantitative analysis MT expression in the tumour cells was observed in 54/54 cases of benign and 32/40 malignant mammary neoplasms. A statistically significant difference at the level of P<0.01 was observed for MT expression between benign and malign mammary tumours in terms of immunoreactivity score. It is concluded that immunohistochemically demonstrated MT expression is significantly associated with benign canine mammary tumours.

Clinicopathological findings in naturally occurring cases of babesiosis caused by large form Babesia from dogs of northeastern Italy.

There are few extensive studies about clinicopathological findings of spontaneous canine babesiosis caused by a large form of the parasite found in Europe. To further characterize and describe clinicopathological findings in dogs affected with this large form of Babesia in northeastern Italy, we evaluated 23 Italian dogs with canine babesiosis by means of clinical history, physical examination, hematological, biochemical, hemostatic tests, serum electrophoresis and urinalysis. Seventeen dogs (74%) had recently traveled on a hunting trip (within 5-15 days of being presented to the clinic) to Bosnia and Herzegovina (n=7), to Croatia (n=8) and to Hungary (n=2). The duration of clinical signs ranged from 1 to 5 days prior to the arrival at the clinic. The main clinical signs were dehydration (100%), apathy (74%), anorexia or decrease appetite (70%) and fever (68%). The anemia was present in 74% of the dogs and classified as mild (35%), moderate (59%) and severe (6%). In all cases, the anemia was normocytic and normochromic. Only three dogs presented erythrocyte regeneration. Seventy percent of dogs had hemolytic anemia and 30% had non-hemolytic anemia. Sixty-nine percent of dogs showed leucopenia and 74% neutropenia. Leucocitosis, due to mature neutrophilia and lymphocytosis, was present in one dog. Activated lymphocytes were noted in 61% of dogs. In all dogs, thrombocytopenia and an elevated hyperfibrinogenemia were present. Significant prolonged activated partial thromboplastin time (aPTT) was only found in one case. In four dogs, both plasma fibrinogen/fibrin degradation products (FDPs) and D-Dimer were increased. Antithrombin (AT) was slightly decreased in 11 of the 23 dogs. In the majority of cases, mild elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatinekinase (CK), total bilirubin and lactic acid and decrease of total iron and total iron binding capacity (TIBC) were present. In conclusion, the main clinicopathological findings were a mild to severe thrombocytopenia, a mild to moderate hemolytic anemia, neutropenia and hyperfibrinogenemia.

Primary gastric histiocytic sarcoma in a dog--a case report.

A 12-year-old intact female mixed breed dog was presented for chronic, intermittent vomiting and diarrhoea. On endoscopic examination a protruding mass arising from the mucosal surface of the pyloric region was detected. Cytological and histological examination revealed an accumulation of pleomorphic round/oval phagocytic cells suggesting histiocytic origin. This was confirmed by immunohistochemistry. No extra-gastric involvement was detected on clinical examination or at necropsy. This is the first report of primary gastric histiocytic sarcoma in a dog.