[show abstract][hide abstract] ABSTRACT: The prevalence of piroplasm (order Piroplasmida) infection was assessed in blood and bone marrow samples from 91 red foxes (Vulpes vulpes) from northern, central and southern Portugal by means of molecular methods. PCR for the 18S rRNA gene of Babesia spp. followed by sequencing revealed 63 foxes positive for the Babesia microti-like piroplasm (syn. Theileria annae) (69.2%; 95% confidence interval [CI]: 58.7-78.5%) and one fox positive for Babesia canis (1.1%; 95% CI: 0.0-6.0%). Positivity to the B. microti-like piroplasm or B. canis in 43 blood samples (83.7%) was significantly higher (p<0.001) than in 43 paired bone marrow samples (20.9%). There were no statistically significant differences in the prevalence of infection between genders (p=0.219) or age groups (<2 years vs. ≥2 years) (p=1.0). This is the first report of the B. microti-like piroplasm in foxes from Portugal as well as the first report on detection by PCR and genotyping of B. canis in a red fox worldwide. A natural cycle of the B. microti-like piroplasm is suggested in red fox populations based on the high prevalence of the protozoan. Red foxes might be a reservoir of the B. microti-like piroplasm and a source of infection to dogs.
[show abstract][hide abstract] ABSTRACT: To determine the prevalence of perinuclear antineutrophil cytoplasmic autoantibodies (pANCA) in dogs with confirmed or suspected immune-mediated hemolytic anemia (IMHA) or dogs infected with various vector-borne pathogens, including Rickettsia rickettsii, Bartonella henselae, Bartonella vinsonii subsp berkhoffii, Ehrlichia canis, Borrelia burgdorferi, and Leishmania infantum.
55 dogs with confirmed or suspected IMHA, 140 dogs seroreactive for vector-borne pathogens, and 62 healthy dogs and dogs seronegative for vector-borne pathogens.
Samples were allocated to subgroups on the basis of the health status of the dogs and the degree of seroreactivity against various vector-borne pathogens. Serum samples were tested retrospectively via indirect immunofluorescence assay to determine pANCA status.
26 of 55 (47%) dogs with confirmed or suspected IMHA and 67 of 140 (48%) dogs seroreactive for vector-borne pathogens had positive results when tested for pANCA. Serum samples with the highest antibody concentrations against L infantum antigen had the highest proportion (28/43 [65%]) that were positive for pANCA. One of 20 (5%) dogs seronegative for tick-borne pathogens and 8 of 22 (36%) dogs seronegative for L infantum had positive results for pANCA. One of 20 (5%) healthy dogs had serum antibodies against pANCA.
pANCA were detected in a high percentage of dogs with IMHA and vector-borne infectious diseases. Therefore, pANCA may be a relatively nonspecific marker for dogs with inflammatory bowel disease, although they could represent a biomarker for immune-mediated diseases and infections.
American Journal of Veterinary Research 09/2012; 73(9):1403-9. · 1.35 Impact Factor
[show abstract][hide abstract] ABSTRACT: Cytologic evaluation plays an important role in the diagnosis of ovarian neoplasia in dogs, but is supported by only scant information on cytologic features of canine ovaries.
The aim of this study was to provide detailed cytologic descriptions of normal canine ovaries in different stages of estrus with comparison to histologic features.
Client-owned dogs admitted for elective ovariohysterectomy were studied. For each dog, fine-needle aspirates of both ovaries were collected, stained, and examined and were compared with histologic sections of the same ovary. The stage of estrus was confirmed by examination of histologic sections and cytologic specimens of vaginal cells collected with swabs.
Thirty-two ovaries from 16 dogs were examined. Luteal cells were observed in 82% of the dogs in diestrus. In early diestrus these cells were polygonal with amphophilic to deeply basophilic cytoplasm, and in late diestrus luteal cells had blebbed cell borders and clear cytoplasm with numerous small vacuoles. Perivascular arrangements and leuko-emperipolesis were noted in both phases of diestrus. Granulosa cells and spindle cells were found in cytologic specimens from most of the ovaries, and blue-gray extracellular material, sometimes associated with granulosa cells, was present. Medium-sized discrete round cells of undetermined origin were observed in some stages of estrus, and structures classified as corpora albicans were noted occasionally.
Knowledge of specific cytologic features of normal canine ovaries is important for identification of pathologic processes in this organ. The novel findings of luteal cell emperipolesis, extracellular material associated with granulosa cells, and round cells of undetermined lineage warrant further study, which may provide new information on canine ovarian structure and function.
[show abstract][hide abstract] ABSTRACT: 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.
[show abstract][hide abstract] ABSTRACT: A male Jack Russell terrier developed bilateral uveitis and glaucoma at 1 year of age. Since the ocular disease was painful and unresponsive to treatment, both globes were enucleated. Microscopical evaluation of one enucleated globe revealed panuveitis, with pigment dispersion and phagocytosis consistent with the ocular lesions of canine Vogt-Koyanagi-Harada (VKH)-like syndrome. Three years later the dog was represented with severe muscle disease and skin lesions. Due to rapid clinical deterioration the dog was humanely destroyed. Necropsy examination revealed lichenoid interface inflammation in the skin and mucous membranes, with pigmentary incontinence consistent with VKH-like syndrome and lymphocytic and histiocytic polymyositis with marked muscle atrophy. Canine VKH-like syndrome is an autoimmune disease that targets melanocyte antigens. Some human patients with VKH disease develop additional autoimmune diseases. To our knowledge this is the first reported case of polymyositis subsequent to VKH-like disease in a dog. In addition, VKH-like disease has not been previously reported in a Jack Russell terrier.
Journal of comparative pathology 12/2010; 144(4):317-23. · 1.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: Mycobacterium celatum is a slow growing non-tuberculous mycobacterium described mainly as occurring in human patients. Only two cases of infection with this pathogen have been reported previously in animals. A 5-year-old, neutered male ferret was presented with progressive weight loss and muscle atrophy. Pale mucous membranes, slight alopecia of the tail and splenomegaly, confirmed by abdominal ultrasound, were observed. Fine-needle aspirations of the spleen revealed extramedullary haematopoiesis and marked macrophage-dominated inflammation associated with mycobacterial infection. Ziehl-Neelsen staining demonstrated sporadic acid-fast bacilli within macrophages. These organisms were identified as M. celatum by microbiological and molecular methods. Phylogenetic analysis based on the 16S rDNA gene compared this isolate with previously reported strains and demonstrated close relatedness to the human strains of M. celatum types 1 and 3. The ferret was treated with enrofloxacin, rifampicin and azithromycin, resulting in clinical improvement. After 40 days of treatment, the spleen was re-evaluated. Cytological evaluation revealed only extramedullary haematopoiesis without evidence of infection. Discontinuation of therapy was followed by rapid deterioration and death.
Journal of comparative pathology 09/2010; 144(2-3):214-8. · 1.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: 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.
[show abstract][hide abstract] ABSTRACT: Canine leishmaniosis (CanL) due to Leishmania infantum is a life threatening zoonotic disease with a wide distribution in four continents and importance also in non-endemic regions. The purpose of this report is to present a consensus of opinions on the diagnosis, treatment, prognosis and prevention of CanL in order to standardize the management of this infection. CanL is a disease in which infection does not equal clinical illness due to the high prevalence of subclinical infection among endemic canine populations. The most useful diagnostic approaches include serology by quantitative techniques and PCR. High antibody levels are associated with severe parasitism and disease and are diagnostic of clinical leishmaniosis. However, the presence of lower antibody levels is not necessarily indicative of disease and further work-up is necessary to confirm CanL by other diagnostic methods such as cytology, histopathology and PCR. We propose a system of four clinical stages, based on clinical signs, clinicopathological abnormalities and serological status. Suitable therapy and expected prognosis are presented for each of the stages. The combination of meglumine antimoniate and allopurinol constitutes the first line pharmaceutical protocol. However, although most dogs recover clinically after therapy, complete elimination of the parasite is usually not achieved and infected dogs may eventually relapse. Follow-up of treated dogs with blood counts, serum biochemistry, urinalysis, serology and PCR is essential for prevention of relapses. Protection against sand fly bites by topical insecticides is effective in reducing infection, and recent development of vaccines has indicated that prevention by vaccination is feasible.
[show abstract][hide abstract] ABSTRACT: A 15-year-old domestic shorthair feline immunodeficiency virus-positive cat was presented with a five day history of productive cough and acute respiratory distress. Physical examination revealed inspiratory dyspnoea and diffuse gingivostomatitis. Radiographs showed an intratracheal mass located at the level of the sixth and the seventh cervical vertebrae. Bronchoscopy revealed a unique intratracheal mass occluding about 85 per cent of the tracheal lumen. The tracheal mass was removed bronchoscopically. A diagnosis of pyogranulomatous inflammation referable to a mycobacterial infection was made based on cytological and histopathological findings. 16S rRNA polymerase chain reaction testing and sequence analysis identified a novel mycobacterial species, likely a slow grower, with 95 per cent identity with Mycobacterium xenopi. To our knowledge, this is the first description of a tracheal mycobacterial granuloma in a cat, and the first time, a mycobacterium with this sequence has been identified.
Journal of Small Animal Practice 04/2009; 50(3):143-6. · 1.18 Impact Factor
[show abstract][hide abstract] ABSTRACT: 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.
[show abstract][hide abstract] ABSTRACT: 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.
Zoonoses and Public Health 10/2008; 55(8-10):521-5. · 2.09 Impact Factor
[show abstract][hide abstract] ABSTRACT: Leishmania infantum, the etiological agent of canine leishmaniosis in the Mediterranean region, is vectored by Phlebotomus spp sandflies, which are active during the warmer months of the year. In order to determine whether seasonality in transmission induces seasonal changes in the prevalence of infection by L. infantum and of parasite-specific immune response, two groups of dogs, one in February (n=37) and another in October (n=42), were studied. Clinical signs compatible with leishmaniosis, as well as presence of microscopic skin lesions in the muzzle were recorded for all dogs. Assays were also performed for detection of L. infantum parasites in muzzle skin samples (PCR, immunohistochemistry and culture), specific serum antibodies (ELISA), and specific lymphocyte proliferation and interferon-gamma production. Although prevalence of non-specific clinical signs increased significantly after the sandfly season, this was not the case for Leishmania-specific markers: positivity by PCR (24% vs. 21%) or immunohistochemistry (3% vs. 2%) of muzzle skin samples, as well as lymphocyte proliferation (59% vs. 50%) or interferon-gamma production (21% vs. 27%) were similar in February and in October. Only prevalence of positive specific antibody titers increased noticeably in October (8% vs. 20%), although this was not statistically significant. Overall, the sandfly season did not have a marked impact on the prevalence L. infantum infection or parasite-specific immune responses analyzed in this study.
[show abstract][hide abstract] ABSTRACT: 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.
[show abstract][hide abstract] ABSTRACT: The aim of this study was to detect Leishmania infantum DNA by real-time PCR in urine from different groups of dogs with clinical leishmaniosis. Urine from 10 clinically healthy dogs and 43 dogs with clinical leishmaniosis diagnosed by positive serology and/or bone marrow PCR were studied. The group of 43 dogs with clinical leishmaniosis was divided into three subgroups: 13 dogs with renal insufficiency and proteinuria (urine protein-creatinine ratio greater than one), 13 dogs with only proteinuria, and 17 dogs with neither renal insufficiency nor proteinuria. The detection of Leishmania DNA was performed by light cycler real-time PCR using hybridization probes in each urine sample. Leishmania positive PCR was found in 47% (20/43) of the urine from leishmaniotic dogs, while all urine from clinically healthy dogs were negative. The percentages of positive Leishmania PCR were 85% (11/13) in dogs with renal insufficiency and proteinuria, 23% (3/13) in dogs with proteinuria and 35% (6/17) in dogs with neither renal insufficiency nor proteinuria. Dogs with renal insufficiency and proteinuria presented a statistical significant greater percentage of positive Leishmania PCR in urine when compared with the other subgroups (P<0.02). This study demonstrates the presence of Leishmania DNA in urine of dogs with leishmaniosis. Those dogs with severe renal damage present a greater number of Leishmania parasites in urine.
[show abstract][hide abstract] ABSTRACT: 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%.
Annals of the New York Academy of Sciences 11/2006; 1078:515-8. · 4.38 Impact Factor
[show abstract][hide abstract] ABSTRACT: 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.
Veterinary Research Communications 10/2006; 30(7):723-33. · 1.08 Impact Factor
[show abstract][hide abstract] ABSTRACT: The aim of the present study is to highlight the advantages of real-time quantitative PCR intended to aid in the diagnosis and monitoring of canine leishmaniosis. Diagnosis of canine leishmaniosis is extremely challenging, especially in endemic areas, due to the diverse and non-specific clinical manifestations, and due to the high seroprevalence rate in sub-clinical dogs. Veterinarian clinicians are usually confronted with cases that are compatible with the disease, and with several diagnostic tests, sometimes with contradictory results. We have developed a new TaqMan assay, targeting the kinetoplast, applied to 44 samples of bone marrow aspirate or peripheral blood. The dynamic range of detection of Leishmania DNA was established in 7 logs and the limit of detection is 0.001 parasites in the PCR reaction. At the time of diagnosis parasitemia ranges from less than 1 to 10(7)parasites/ml. The ability to quantify the parasite burden allowed: (i) to elucidate the status of positive dogs by conventional PCR, although larger studies are necessary to clarify the dividing line between infection and disease, (ii) to estimate the kinetics of the parasite load and the different response to the treatment in a follow-up and (iii) to validate blood as less invasive sample for qPCR. The continuous data provided by real-time qPCR could solve the dilemma for the clinician managing cases of canine leishmaniosis by differentiating between Leishmania-infected dogs or dogs with active disease of leishmaniosis.