A comparison of cytologic quality in fine-needle specimens obtained with and without aspiration from superficial lymph nodes in the dog.

OBJECTIVES: To assess the impact of aspiration on cytologic quality of fine-needle specimens obtained from lymph nodes of dogs and to compare the level of agreement of the cytologic diagnosis between the two sampling techniques. MATERIALS AND METHODS: Fifty-three client-owned dogs were prospectively enrolled. Client-owned dogs were prospectively enrolled in the study if cytologic examination of lymph nodes was indicated in the course of their clinical diagnostic work-up. In each dog, two superficial, palpably accessible lymph nodes were sampled: one by fine-needle aspiration and the other by fine-needle non-aspiration, using a 21-Gauge needle with or without a 5-mL syringe. Cytologic quality was assessed in duplicate smears assessed by two observers who rated cellularity, blood contamination, thickness, cell preservation and cytoplasmic fragmentation using a predefined scoring system. RESULTS: Fifty-three client-owned dogs were included in the study. No significant difference was found in rating scores between the two sampling techniques for any of the cytologic quality parameters assessed. Cohen's kappa coefficient was 0.84 (95% confidence interval 0.68-1.00), indicating diagnostic agreement between the sampling techniques. CLINICAL SIGNIFICANCE: In this study, both fine-needle aspiration and fine-needle non-aspiration techniques yielded lymph node specimens of comparable cytologic quality and with acceptable agreement in cytologic diagnosis.

A prospective epidemiological, clinical, and clinicopathologic study of feline leukemia virus and feline immunodeficiency virus infection in 435 cats from Greece.

Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) are retroviruses causing significant morbidity and mortality in cats. The aim of this study was to describe the epidemiological, clinical and clinicopathologic aspects of FeLV and FIV infections in different populations of cats in Greece, including client-owned cats, stray cats and cats who live in catteries. A total of 435 cats were prospectively enrolled. Serological detection of FeLV antigen and FIV antibody was performed using a commercial in-house ELISA test kit. The results showed that 17 (3.9 %) and 40 (9.2 %) of the 435 cats were positive for FeLV antigen and FIV antibody, respectively, whereas 5 (1.1 %) had concurrent infection with FeLV and FIV. Factors that were associated with FeLV antigenemia, based on multivariate analysis, included vomiting, rhinitis, infection with FIV, neutropenia, decreased blood urea nitrogen and increased serum cholesterol and triglyceride concentrations. Factors associated with FIV seropositivity included male gender, older age, outdoor access, weight loss, fever, gingivostomatitis, skin lesions and/or pruritus and hyperglobulinemia. Various clinical signs and laboratory abnormalities were found to be significantly associated with retroviral infections, suggesting that current guidelines to test all sick cats should be followed, taking into particular consideration the high-risk groups of cats found in this study.

First report of fatal autochthonous angiostrongylosis in a dog in Greece.

Canine angiostrongylosis caused by Angiostrongylus vasorum is a life-threatening disease which is emerging in regions of Europe. Thus, there is the merit for a continuous epidemiological surveillance in dog populations. This is the first description of a clinical autochthonous case of canine angiostrongylosis in Greece. A 7-month-old, male, mixed-breed dog was presented with progressively worsening anorexia, respiratory distress, coughing, bleeding diathesis and succumbed four days post admission. Gross post mortem examination revealed numerous nodular fistulated lesions in the lungs, and pulmonary cytology and histopathology showed a verminous pyogranulomatous pneumonia. The definitive diagnosis was based on the morphological identification of first stage larvae (L1) retrieved in the feces by the Baermann method, the detection of the circulating antigen by an in-clinic test and the molecular identification of L1. This report underlines the epidemiological and clinical implications, as well as the infection risks when the index of clinical suspicion is low and the disease is unexpected in a country.

Canine pancytopoenia in a Mediterranean region: a retrospective study of 119 cases (2005 to 2013).

OBJECTIVES: To further clarify the causes of pancytopoenia and to investigate whether underlying cause or severity were associated with survival in an area endemic for vector-borne pathogens. METHODS: Retrospective review of medical records of 119 dogs with and 238 dogs without pancytopoenia. RESULTS: Mixed-breed dogs and dogs younger than one year had higher odds of being pancytopoenic. The most common diagnoses included monocytic ehrlichiosis (n=42), leishmaniasis (n=28) and parvoviral enteritis (n=19). The mean white blood cell counts were lower in dogs with ehrlichiosis and parvoviral enteritis compared to dogs with leishmaniasis, while platelet counts were lower in ehrlichiosis compared to leishmaniasis or parvoviral enteritis. Total protein concentrations were lower in dogs with parvoviral enteritis compared to ehrlichiosis and leishmaniasis. Higher haematocrit, platelet and white cell counts were associated with better odds of survival. CLINICAL SIGNIFICANCE: Infectious diseases appear to be the leading causes of canine pancytopoenia in endemic areas; severe leukopoenia (ehrlichiosis, parvoviral enteritis), thrombocytopoenia (ehrlichiosis) and hypoproteinaemia (parvoviral enteritis), represented potentially useful disease-specific diagnostic determinants. The severity of pancytopoenia significantly affects the clinical outcome.

Serum acute phase proteins as clinical phase indicators and outcome predictors in naturally occurring canine monocytic ehrlichiosis.

BACKGROUND: Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, is an important tick-borne disease of global importance. Currently, limited information is available on the diagnostic and prognostic value of acute phase proteins (APPs) in dogs naturally infected with E. canis. HYPOTHESIS: APPs may be useful indicators of the clinical phase of CME and predictive of the clinical outcome (death or survival). ANIMALS: Fifty-six dogs naturally infected with E. canis and 7 clinically healthy control dogs. METHODS: C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), and albumin concentrations determined on admission were retrospectively compared among 27 dogs with nonmyelosuppressive CME, 29 dogs with myelosuppressive CME and 7 healthy dogs. Diagnosis of CME was based on clinical and clinicopathological findings, seropositivity to E. canis, polymerase chain reaction amplification of E. canis-specific 16S rDNA, microscopic observation of Ehrlichia sp. morulae in blood monocytes or some combination of these. RESULTS: Mean concentrations of CRP, SAA, and Hp were significantly higher in the myelosuppressed dogs compared with the other groups, but no significant differences were found in the concentration of albumin. Survival analysis of the affected animals indicated that APP concentrations were not associated with clinical outcome; the latter was strongly associated with pancytopenia (odds ratio for death 22.7) and neutropenia (odds ratio for death 7.7). CONCLUSIONS AND CLINICAL IMPORTANCE: CRP, SAA, and Hp serum concentrations on admission are useful indicators of the clinical phase of CME, but are not useful predictors of clinical outcome.

Absence of myelofibrosis in dogs with myelosuppression induced by Ehrlichia canis infection.

Bone marrow (BM) pathology was assessed in 10 dogs with Ehrlichia canis-induced aplastic pancytopenia. BM core biopsy sections were stained with haematoxylin and eosin and with haematoxylin/van Gieson and Gordon and Sweets' reticulin stain for the detection of collagen and reticulin fibres, respectively. Iron stores were assessed by Perls' Prussian blue staining. There was no significant deposition of collagen or reticulin in any sample, but in seven dogs the BM was depleted of haemosiderin. These findings suggest that myelofibrosis does not play a significant role in the development of BM failure in canine monocytic ehrlichiosis and that iron deficiency may exacerbate the anaemia in the myelosuppressive phase of the disease.

Molecular documentation of Bartonella infection in dogs in Greece and Italy.

This study reports the occurrence of "Bartonella rochalimae" in Europe and the presence of Bartonella vinsonii subsp. berkhoffii genotypes II and III in dogs in southern Italy and provides DNA sequencing evidence of a potentially new Bartonella sp. infecting dogs in Greece and Italy.

Factors associated with the occurrence of epistaxis in natural canine leishmaniasis (Leishmania infantum).

BACKGROUND: Canine leishmaniasis (CanL) is a common cause of epistaxis in dogs residing in endemic areas. The pathogenesis of CanL-associated epistaxis has not been fully explored because of the limited number of cases reported so far. HYPOTHESIS: Epistaxis in CanL could be attributed to more than 1 pathomechanism such as hemostatic dysfunction, biochemical abnormalities, chronic rhinitis, and coinfections occurring in various combinations. ANIMALS: Fifty-one dogs with natural CanL. METHODS: The allocation of 51 dogs in this cross-sectional study was based on the presence (n = 24) or absence (n = 27) of epistaxis. The potential associations among epistaxis and concurrent infections (Ehrlichia canis, Bartonella spp., and Aspergillus spp.), biochemical and hemostatic abnormalities, and nasal histopathology were investigated. RESULTS: Hypergammaglobulinemia (P= .044), increased serum viscosity (P= .038), decreased platelet aggregation response to collagen (P= .042), and nasal mucosa ulceration (P= .039) were more common in the dogs with epistaxis than in those without epistaxis. The other significant differences between the 2 groups involved total serum protein (P= .029) and gamma-globulin (P= .013) concentrations, which were higher, and the percentage platelet aggregation to collagen, which was lower (P= .012) in the epistaxis dogs. CLINICAL IMPORTANCE: CanL-associated epistaxis appears to be the result of multiple and variable pathogenetic factors such as thrombocytopathy, hyperglobulinemia-induced serum hyperviscosity, and nasal mucosa ulceration.

Acute myelomonocytic leukaemia with short-term spontaneous remission in a cat.

A 2-year-old, spayed female domestic shorthair cat was referred with a history of anorexia and depression of 1 week duration. On physical examination, the cat was lethargic and febrile, with splenomegaly, anisocoria and ulcerative stomatitis. A complete blood count (CBC) and a biochemistry profile showed leukocytosis, numerous blast cells in the peripheral blood, thrombocytopenia, hyperglobulinaemia and a positive test for feline leukaemia virus antigen. A diagnosis of acute myelomonocytic leukaemia was made on the basis of the results of bone marrow cytology, histopathology, and immunochemistry (CD3, CD79a, lysozyme, and myeloperoxidase) tests. Following an unexpected 1-month period of clinical and clinicopathological remission without chemotherapy, the cat relapsed and died 1 week later.

A retrospective study of 61 cases of spontaneous canine epistaxis (1998 to 2001).

OBJECTIVES: To determine the prevalence and identify possible clinicopathologic indicators of the diseases associated with canine epistaxis. METHODS: The medical records of 61 dogs with epistaxis were reviewed. RESULTS: Systemic diseases, diagnosed in fifty-six dogs, included canine leishmaniasis in twenty-three dogs, canine monocytic ehrlichiosis in twenty-two, concurrent canine leishmaniasis and canine monocytic ehrlichiosis in six, rodenticide toxicity in two and primary immune-mediated thrombocytopenia, suspected oestrogen toxicity and systemic arterial hypertension in one dog each. Intranasal diseases were documented in the remaining five dogs, including transmissible venereal tumour in three dogs, and nasal adenocarcinoma and nasal aspergillosis in one dog each. Mucosal pallor and a generalised bleeding tendency were significantly more common among dogs with canine monocytic ehrlichiosis compared with those with canine leishmaniasis, whereas the opposite was true for peripheral lymphadenomegaly. Also, dogs with canine monocytic ehrlichiosis presented with pancytopenia more frequently compared with those with canine leishmaniasis; in the latter dogs, the median values of haematocrit, leucocyte and platelet counts and serum total protein concentrations were higher. CLINICAL SIGNIFICANCE: Canine leishmaniasis and canine monocytic ehrlichiosis are the leading causes of canine epistaxis in Greece. Mucosal pallor, bleeding tendency and pancytopenia are more likely to be indicative of canine monocytic ehrlichiosis, as opposed to peripheral lymphadenomegaly and hyperproteinaemia in canine leishmaniasis.

Acute tumour lysis syndrome in a dog with B-Cell multicentric lymphoma.

A 5-year-old, spayed female German Shepherd dog was admitted to hospital with marked generalised lymphadenomegaly and splenomegaly. A stage Va B-cell multicentric lymphoma was diagnosed on clinical, cytological (lymph node, bone marrow), histological-immunohistochemical (lymph node excision) and imaging grounds. Since no satisfactory remission was achieved using a multi-drug chemotherapy protocol that included cyclophosphamide, vincristine, cytosine arabinoside, prednisolone, and subsequently supplemented by L-asparaginase, it was replaced by another protocol combining vincristine, L-asparaginase, prednisolone, cyclophosphamide and doxorubicin. Soon after the third weekly session of the second protocol, the clinical status of the animal deteriorated suddenly and severely, with a bleeding tendency, jaundice, hyperuricaemia, hyperphosphataemia, azotaemia, hyperbilirubinaemia and, presumptive disseminated intravascular coagulation. There was also complete regression of lymphadenomegaly. This report emphasises the clinicopathological features and the diagnostic peculiarities of the acute tumour lysis syndrome, which occurs uncommonly in dogs.

Comparative methodology for the detection and differentiation of circulating microfilariae of Dirofilaria immitis in the dog.

The sensitivities of the Knott's test (four 20-microl sediment aliquots), quantitative buffy coat capillary tube method (QBC tube, 111 microl of whole blood) and direct blood smear (DBS, 20 microl of whole blood) were evaluated for the detection of microfilaraemia in dogs. Undiluted whole blood samples taken from 70 Dirofilaria immitis antigen-positive dogs and 10 serially diluted microfilaraemic blood samples at concentrations of 400, 200, 100, 50, 25 and 12 microfilariae (mff) ml(-1) were examined. For filarial speciation, the buffy coat of QBC tubes was mixed with one drop of methylene blue-formalin solution and examined as a direct smear. In 52/70 microfilaraemic blood samples, the number of mff ranged from 12 to 321987 ml(-1) (median: 3199 ml(-1)). The diagnostic sensitivity of the Knott's test, QBC tube method and DBS in undiluted blood samples attained the 100%, 98% and 92.3% levels, respectively. Eighteen dogs tested amicrofilaraemic by all three methods. At concentrations of 400 mff ml(-1), a 100% sensitivity was found by all three methods, while at 200 mff ml(-1) the Knott's test, QBC tube and DBS were 100%, 100% and 90% sensitive, respectively. The relevant figures at 100 mff ml(-1) were 100%, 100% and 80%, at 50 mff ml(-1) 100%, 100% and 50%, at 25 mff ml(-1) 100%, 100% and 10% and at 12 mff ml(-1) 80%, 50% and 10%. At 50 and 25 mff ml(-1), the DBS was less sensitive compared to the other two methods, while at 12 mff ml(-1), only to the Knott's test. A significant correlation was found between the QBC tube method and Knott's test regarding mff speciation. Therefore, the QBC method may be considered a reliable alternative to the Knott's test for both the detection and speciation of mff in the dog.

Evaluation of cytology in the diagnosis of acute canine monocytic ehrlichiosis (Ehrlichia canis): a comparison between five methods.

The purpose of this study was the comparison of the diagnostic sensitivity between buffy coat (BC), peripheral blood (PB), lymph node (LN), bone marrow (BM) and short-term culture (P-D) cytology that has been based on the detection of Ehrlichia canis morulae, in the acute phase of canine monocytic ehrlichiosis (CME). Their cellular localization, total numbers and microscopic differentials were also investigated. The highest sensitivities were achieved after evaluating 1000 oil immersion fields (OIFs) in BC (66%) and an equal number in LN (60.9%) smears, separately or together (74%). The morulae were more often detected into lymphocytes than monocytes. The highest total number of morulae (n=143) were found in P-D smears. Finally, to avoid false positive diagnoses, platelets, lymphocytic azurophilic granules, lymphoglandular bodies and phagocytosed nuclear material should not be confused with the morulae.

Protothecal colitis in a German shepherd dog.

A 3 1/2-year-old intact male German Shepherd Dog was admitted with a history of protracted haemorrhagic diarrhoea and tenesmus. Clinical evaluation revealed thickening of the rectal mucosa and presence of fresh blood and mucus in the faeces. Severe mucosal granularity and multiple nodules, in addition to a rectal stricture 7 to 9 cm from the anus, were found on colonoscopy. Histopathological evaluation of biopsy specimens identified organisms with morphologic features compatible with those reported for Prototheca spp. Although surgical excision of the rectal stricture was performed and ketoconazole treatment was instituted postoperatively, the dog's clinical condition deteriorated and euthanasia was performed 4 months later.

A randomised, blinded, placebo-controlled clinical trial with allopurinol in canine leishmaniosis.

A total of 45 non-uremic dogs, with clinical signs indicating leishmaniosis, entered the study. Diagnosis was confirmed by indirect immunofluorescence assay (IFA) on serum and polymerase chain reaction (PCR) on bone marrow samples. The dogs were randomly allocated into Group A (n=37) that received allopurinol (10mg/kg B.W., per os, twice daily) for 4 consecutive months, and Group B (n=8) that were placebo-treated. Clinical signs were scored just before and at monthly intervals throughout the study period, in a blinded and independent fashion. Complete blood count, serum biochemistry profile, urinalysis, lymph node and bone marrow parasitology, IFA and enzyme-linked immunosorbent assay (ELISA) serology and bone marrow PCR were carried out at the beginning and at the end of the trial. A total of three Group A and one Group B dogs died of end stage kidney disease that developed during the trial. In Group A animals that endured the trial there was a significant improvement in the general body condition, conjunctivitis, peripheral lymphadenopathy, splenomegaly, masticatory muscle atrophy, ulcerative stomatitis, epistaxis, exfoliative dermatitis, cutaneous ulcerations, blepharitis and nasodigital hyperkeratosis. The same observation was made for anemia, lymphopenia, hyperproteinemia, hyperglobulinemia, hyperphosphatemia, increased alkaline phosphatase activity and the low albumin/globulin ratio. By contrast, no improvement of any kind was seen in Group B dogs. Lymph node and bone marrow parasite numbers were significantly decreased in Group A animals. In Group B, that occurred only in the lymph nodes. Apart from remission of clinical signs and restoration to normal of clinicopathological abnormalities, allopurinol did not eliminate Leishmania organisms, as the PCR result on bone marrow was still positive in all the dogs that finished the trial.