Investigation of comorbidities in dogs with leishmaniosis due to Leishmania infantum.

In endemic areas, dogs with leishmaniosis due to Leishmania infantum frequently have comorbidities, including mostly neoplastic, infectious, and parasitic diseases. The aim of this study was to compare the prevalence of comorbidities among dogs that are not infected by L. infantum, dogs that are infected but do not present leishmaniosis, and dogs with leishmaniosis, and to examine if certain comorbidities are independent risk factors for the infection by L. infantum and/or for the development of canine leishmaniosis (CanL). A total of 111 dogs, older than 1-year and non-vaccinated against CanL, were allocated into three groups: group A (n = 18) included dogs that were not infected by L. infantum, group B (n = 52) included dogs that were infected by L. infantum but did not present CanL, and group C (n = 41) included dogs with CanL. Signalment and historical data were obtained using a structured questionnaire. Laboratory examinations included complete blood count, serum biochemistry, urinalysis, fecal parasitology, modified Knott's test, microscopic examination of capillary blood, buffy coat, lymph node, bone marrow and conjunctival smears, qualitative serology for Dirofilaria immitis, Anaplasma phagocytophilum/A. platys, Borrelia burgdorferi and E. canis, IFAT for L. infantum, ELISA for Babesia spp. and Neospora caninum, and real-time PCR for L. infantum in bone marrow, skin biopsies and conjunctival swabs. A variety of comorbidities were found in all three groups. No independent risk factors for infection by L. infantum were found. On the contrary, among dogs infected by L. infantum, being a mongrel [odds ratio (OR): 11.2], not receiving prevention for dirofilariosis (OR: 26.5) and being seropositive to N. caninum (OR: 17.1) or to Babesia spp. (OR: 37.6), were independent risk factors for presenting CanL. Although no comorbidities influence the probability of canine infection by L. infantum, certain comorbidities may be precipitating factors for the transition from the subclinical infection by L. infantum to the overt CanL.

Report on the First African Swine Fever Case in Greece.

African swine fever (ASF) poses a major threat to swine health and welfare worldwide. After several European countries have reported cases of ASF, Greece confirmed officially the first positive case on 5 February 2020. The owner of a backyard farm in Nikoklia, a village in Serres regional unit, Central Macedonia, reported a loss of appetite, weakness, dyspnea, and the sudden death of 6 domestic pigs. Necropsy was performed in one gilt and findings were compatible with acute to subacute septicemic disease. Predominantly, hyperemic enlargement of spleen and lymph node enlargement and/or hemorrhage were observed. Description of vague clinical signs by the farmer suggested a limited resemblance to ASF-acute infection. However, the disease could not be ruled out once septicemic condition including splenomegaly, was diagnosed macroscopically at necropsy. In addition, considering the farm's location near to ASF protection zones, a further diagnostic investigation followed. Confirmation of the disease was obtained using a series of diagnostic tests on several tissue samples. Further clinical, molecular, and epidemiologic evaluation of the farm was performed. According to the contingency plan, authorities euthanized all 31 pigs on the farm, whilst blood testing revealed ASF virus infection. Further emergency measures were implemented to contain the spread of the disease.

Evaluation of serum symmetric dimethylarginine as a biomarker of kidney disease in canine leishmaniosis due to Leishmania infantum.

Canine leishmaniosis (CanL)-associated chronic kidney disease is a leading cause of morbidity and mortality in Mediterranean countries. Novel renal biomarkers, such as serum symmetric dimethylarginine (sSDMA), may be useful surrogates for the detection of renal functional impairment. The objectives of this study were to investigate sSDMA concentrations in dogs with CanL, with and without azotemia, and to establish any potential association with the prevalence and severity of proteinuria, with the prevalence of decreased urine specific gravity and with the LeishVet clinical stages of CanL. Serum samples from 68 dogs with CanL (50 nonazotemic and 18 azotemic) and 17 healthy dogs were retrospectively examined. Increased sSDMA was documented in 26 % of dogs with CanL without azotemia and in 83.3 % of dogs with azotemia. Serum SDMA was significantly higher in azotemic compared to nonazotemic dogs and was associated with the presence and severity of proteinuria, the decreased urine specific gravity and the advanced clinical stages of CanL. The results of the present study indicate that sSDMA may be a useful adjunct to serum creatinine and urine protein/creatinine ratio for the detection of CanL-associated nephropathy, but it is of limited value for distinguishing among the LeishVet clinical stages of CanL.

Factors affecting the occurrence, duration of hospitalization and final outcome in canine parvovirus infection.

The objectives of this matched case-control study in a veterinary teaching hospital were to investigate the influence of signalment and historical data on the odds of occurrence of canine parvovirus (CPV) enteritis and the potential usefulness of the clinical signs and clinicopathologic abnormalities recorded on admission as prognostic indicators of mean duration of hospitalization (DOH) and outcome of the disease. Ninety-four puppies with natural CPV enteritis and 188 age-matched controls were studied. The odds to develop CPV enteritis were higher in purebreds compared to mixed-breed puppies. Vomiting and depression at the time of admission were associated with a prolongation of DOH by 2 and 1.75 days, respectively. The lymphopenic and hypoalbuminemic dogs were hospitalized for 1.9 and 2.5 more days, respectively, compared to those without these abnormalities. The odds of non-survival were higher in those puppies with evidence of systemic inflammatory response syndrome (SIRS) at the time of admission.

Comparison between ultrasonographic and caliper measurements of testicular volume in the dog.

The aim of this study was to compare the accuracy of two methods used to clinically estimate testicular volume in the dog. Caliper and ultrasonographic measurements of testicular dimensions (length, width and height) of both testes were performed on 21 adult Beagles. Either measurement was taken in vivo over the scrotal skin and again in vitro after castration, excluding or including, when possible, the epididymis. In a sub-sample of 14 testes of 7 randomly selected dogs, the testicular volume was calculated by the formula of an ellipsoid: length (l)xwidth (w)xheight (h)x0.5236, and the empiric formula of Lambert: (l)x(w)x(h)x0.71. The calculated volumes were then compared to the actual ones, which were estimated by water displacement. For each measurement method, the formula that estimated more accurately the true testicular volume was, subsequently, applied to the calculation of the testicular volume of the remaining testes. The formula of ellipsoid estimated testicular volume more accurately when in vivo and in vitro caliper measurements and in vitro ultrasound measurements were performed. The formula of Lambert estimated testicular volume more accurately when in vivo ultrasound measurements were performed. The volumes calculated from the measurements of calipers over the scrotal skin overestimated true volumes, on average, by 69+/-27% (concordance correlation coefficient r(ccc)=0.49). By contrast, those calculated from the ultrasonography measurements were more accurate, overestimating the true volumes by 17+/-24% (rccc=0.81). Therefore, ultrasonography when the formula of Lambert is used is more accurate than caliper methods for the estimation of testicular volume in clinical practice and should be the method of choice.

Aetiology of canine otitis externa: a retrospective study of 100 cases.

The objective of this retrospective study was to investigate in 100 dogs with otitis externa (OE) the possible associations between signalment, history, clinical and laboratory findings and the various primary, secondary and perpetuating causative factors of ear canal inflammation. The age of the dogs ranged from 3 months to 14 years (median: 4.75 years) and they included 45 males and 55 females. Cocker spaniels, Jura des Alpes and Brittany spaniels were significantly overrepresented among dogs with OE when compared to the hospital canine population. In the majority of the cases, OE was chronic-recurrent (63%) or bilateral (93%). Allergic dermatitis (43/100 dogs), grass awns (12/100) and otoacariasis (7/100) were the most common primary causative factors; no primary factor could be incriminated in 32 cases and more than one was found in three dogs. Malassezia spp. (66/100 dogs), cocci (38/100) and rods (22/100) were the secondary causative factors, while ear canal stenosis (38/100) and tympanic membrane perforation-otitis media (25/100) were the most important perpetuating factors. Atopic dermatitis and adverse food reactions-associated OE was more common in females and dogs with a history of pruritic skin disease, while grass awn-induced OE occurred in cocker spaniels and acute cases. Tympanic membrane perforation was less frequent in atopic dermatitis and adverse food reactions-associated OE, but more common when otoscopic and ear canal cytological examination revealed the presence of grass awns and rods, respectively. Finally, cocci overgrowth was positively associated with ear canal stenosis.

Relative sensitivity of hair pluckings and exudate microscopy for the diagnosis of canine demodicosis.

This study was designed to compare the sensitivity of deep skin scraping, hair plucking, and exudate microscopy for the diagnosis of canine demodicosis. Sixty-seven dogs diagnosed with demodicosis were enrolled in the study. Thirty dogs had localized and 37 had generalized demodicosis. Twenty-seven of the 67 dogs had complicated (secondarily infected) and 40 had noncomplicated form. On each dog, a single lesion was randomly selected to obtain one deep skin scraping, hair plucking, and, when present (n = 13) exudate. For skin scraping and exudate microscopy, an area under a cover slip measuring 2.2 x 2.2 mm was examined, while trichography included the evaluation of 100 hair shafts. At least one parasitic element was found in 85.1% of trichograms, and 100% of exudate preparations. The number of parasitic elements was higher in skin scrapings compared to the other two methods. The diagnostic sensitivity of skin scrapings was higher than that of hair pluckings for the total number of samples (P = 0.002) and for those obtained from dogs with the localized (P = 0.004) and the noncomplicated (P = 0.002) forms of the disease. The diagnostic sensitivity of hair pluckings was higher in generalized and complicated demodicosis compared to the localized and noncomplicated variants. Based on these results, exudate microscopy may be equally sensitive to deep skin scrapings, and trichography may be of value in generalized and complicated demodicosis, although a negative result cannot rule it out.

Bone marrow mastocytosis in dogs with myelosuppressive monocytic ehrlichiosis (Ehrlichia canis): a retrospective study.

BACKGROUND: Bone marrow mastocytosis has been reported rarely in naturally occurring canine monocytic ehrlichiosis (CME). OBJECTIVES: The aims of the present study were to estimate the prevalence and magnitude of bone marrow mastocytosis in a case series of dogs with natural CME and to assess the association, if any, between mastocytosis and the clinical severity of the disease. METHODS: Seventy-six dogs with confirmed CME (Ehrlichia canis) were included in the study. Affected dogs were allocated into group A (n = 51) without bone marrow hypoplasia and group B (n = 25) with bone marrow hypoplasia. Twenty clinically healthy Beagles not previously exposed to E canis served as controls (group C). The main inclusion criteria for group A were documentation of normocellular to hypercellular bone marrow and complete clinical cure following a 4-week treatment with doxycycline, while those for group B were bone marrow hypoplasia and lack of response to doxycycline. Bone marrow aspirate smears from all 96 dogs were Giemsa-stained and examined for the presence of mast cells, which were calculated as a percentage of 1,000 nucleated cells (NCs). The prevalence of mastocytosis was compared among the 3 groups by the Pearson's chi-square test. RESULTS: Bone marrow mastocytosis (>0.1% of NCs) was found in 5 (20%) dogs in group B (range, 0.5-2.5% of NCs; median, 1% of NCs). One dog in each of groups A and C had 0.1% mast cells in the marrow. The prevalence of bone marrow mastocytosis in dogs in group B was significantly higher (P = .004) than in groups A and C. CONCLUSION: Bone marrow mastocytosis can be seen in a substantial number of dogs with E canis-induced myelosuppression.

Clinical signs and clinicopathologic abnormalities in dogs with clinical spirocercosis: 39 cases (1996-2004).

OBJECTIVE: To determine clinical signs and clinicopathologic abnormalities in dogs with naturally occurring clinical spirocercosis. DESIGN: Retrospective case series. ANIMALS: 39 dogs with spirocercosis. PROCEDURES: Medical records were reviewed, and information on signalment, residence (rural vs urban), owner complaints, physical examination findings, clinicopathologic abnormalities, radiographic and endoscopic findings, and concurrent systemic diseases was recorded. RESULTS: Hellenic hounds and mixed-breed dogs were overrepresented, compared with a group of 117 control dogs without spirocercosis that were examined because of gastrointestinal tract disease, and mean body weight of dogs with spirocercosis was significantly higher than mean body weight of control dogs. Odynophagia (34 [87%]), regurgitation (24 [62%]), and excessive salivation (14 [36%]) were the most common clinical findings. The most common radiographic abnormalities were a mass in the caudodorsal aspect of the mediastinum (15/35 [43%]) and spondylitis of the caudal thoracic vertebrae (10 [29%]). Parasitic nodules were seen during esophagoscopy in all 39 dogs. Normocytic, normochromic, nonregenerative anemia; neutrophilic leukocytosis; hyperproteinemia; and high alkaline phosphatase activity were significantly more common in dogs with spirocercosis than in a control group of 56 healthy dogs. Concurrent systemic diseases, mainly leishmaniosis, dirofilariosis, and monocytic ehrlichiosis, were documented in 14 (36%) dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that clinical spirocercosis occurs more often in young-adult, large-breed dogs. Nonregenerative anemia, neutrophilic leukocytosis, hyperproteinemia, and high alkaline phosphatase activity may be useful clinicopathologic indicators of this disease.

Type of smear may influence thrombopoietic cell counts in the bone marrow of clinically healthy dogs.

BACKGROUND: The expected number of thrombopoietic cells in normal canine bone marrow is poorly defined and there is no consensus on the most appropriate way to prepare cytologic smears to evaluate these cells nor on the optimum method for their quantification. OBJECTIVE: The purpose of this study was to determine total and differential counts of thrombopoietic cells in the bone marrow of clinically healthy Beagle dogs by comparing 4 different smear types and bone marrow core biopsies. METHODS: Twenty-two clinically healthy, male Beagle dogs, 10 to 12 months old, were used in the study. Following bone marrow aspiration and core biopsy from the iliac crest, Giemsa-stained smears were prepared by 4 techniques: drop-squash, particle-squash, buffy coat, and fat-layer smears. Thrombopoietic cells were counted in up to 100 low-power fields (LPF, X10 objective) in the aspiration smears and in all possible high-power fields (HPF, X40 objective) in H&E-stained biopsy sections. RESULTS: Mean total thrombopoietic cell counts were 2.76 cells/LPF (drop-squash), 1.55 cells/LPF (particle-squash), 8.05 cells/LPF (buffy coat), and 3.08 cells/LPF (fat-layer). Core biopsies yielded 5.31 cells/HPF but frequently failed to provide interpretable specimens. There was a significant difference in cell counts among the 4 smear types (P <.001). Based on evaluation of buffy coat smears, thrombopoietic cells included 1.23% megakaryoblasts, 8.77% promegakaryocytes, and 90% megakaryocytes, with a mean maturation index of 0.11. CONCLUSION: Thrombopoietic cell counts in canine bone marrow are influenced by the smear technique. Buffy coat and fat-layer smears may be useful to obtain cellular smears in hemodiluted or small aspirate samples.

Evaluation of lymph node and bone marrow cytology in the diagnosis of canine leishmaniasis (Leishmania infantum) in symptomatic and asymptomatic dogs.

The sensitivity and specificity of lymph node and bone marrow smear microscopy for the diagnosis of Leishmania infantum-infected dogs was evaluated in 79 dogs with leishmaniasis (Group A), 52 asymptomatically infected dogs (Group B), and 44 healthy noninfected dogs (Group C). Light microscopy examination included 10 to 1,000 oil immersion fields, and the density of Leishmania amastigotes was scored by a 0 to +6 scale. Using polymerase chain reaction as the gold standard, the specificity of lymph node and bone marrow cytology was 100%, whereas sensitivity ranged from 7.8% to 92.6%, being significantly higher in Group A compared with Group B. The amastigote scores were also significantly higher in Group A compared with Group B. These results indicate that lymph node and bone marrow cytology is a highly sensitive and specific method for the diagnosis of canine patent leishmaniasis, whereas its sensitivity is relatively low in asymptomatic infections.

Periodic administration of allopurinol is not effective for the prevention of canine leishmaniosis (Leishmania infantum) in the endemic areas.

A total of 95 clinically healthy and seronegative for Leishmania infantum dogs, residing an area highly endemic for canine leishmaniosis (CL) and living an outdoor life-style, were split into positive and negative groups, and then were randomly assigned to receive allopurinol (n=51; 20 mg/kg once daily), or placebo (n=44) for 1 week per month, from April to November. Forty per cent (38/95) of these dogs were not reexamined and retested at the end of the trial for reasons unrelated to CL. None of the remaining 57 dogs exhibited the symptomatic form of the disease at the end of the 1-year follow-up period. Of the 15 allopurinol-treated dogs that were non-infected (negative PCR and tissue smear microscopy) at the beginning of the trial, 6 (40% P=0.03) became PCR-positive, of which 3 became also seropositive, at the end of the observation period. In contrast, only 1 of 7 (14.3%) placebo-treated non-infected dogs became PCR positive at the same time point. Of the 19 allopurinol-treated dogs that were infected (PCR-positive) at the beginning of the trial, 18 (94.7%) remained PCR-positive and one (5.3%) seroconverted, at the end of the observation period. Of the 16 initially infected and placebo-treated dogs, 14 (87.5%) remained PCR positive, of which one (6.7%) also turned positive by tissue smear microscopy. Therefore, it is concluded that the use of allopurinol, at the daily dose of 20mg/kg, for 1 week per month, during the period of sandfly activity, does not prevent the infection of non-infected dogs by L. infantum, and, also, does not help in the elimination of the parasite from dogs with asymptomatic infections.

Chronic canine ehrlichiosis (Ehrlichia canis): a retrospective study of 19 natural cases.

Nineteen dogs from Greece with chronic ehrlichiosis were studied. The dogs exhibited bicytopenia or pancytopenia, bone marrow hypoplasia, seroreactivity to Ehrlichia canis (E. canis) antigens, and had no history of drug or radiation exposure. Anorexia, depression, severe bleeding tendencies, hypoalbuminemia, and increased serum alanine aminotransferase activity were also hallmarks of the disease. All these animals eventually died, irrespective of the treatment applied. Some dogs were also serologically positive for Rickettsia conorii, Leishmania infantum (L. infantum), and Bartonella vinsonii subspp. berkhoffii. Polymerase chain reaction testing of bone marrow samples revealed E. canis, Anaplasma phagocytophilia, Anaplasma platys, and L. infantum in some dogs. Concurrent infections did not appear to substantially influence the clinical course and final outcome of the chronic canine ehrlichiosis.

A cross-sectional study of Leishmania spp. infection in clinically healthy dogs with polymerase chain reaction and serology in Greece.

A total of 73 clinically healthy hunting dogs, experiencing an outdoor lifestyle and originating from an area where canine leishmaniasis is endemic, were included in the study. Polymerase chain reaction (PCR) and indirect immunofluorescent antibody test (IFAT) for Leishmania spp. were done on bone marrow and serum samples, respectively, obtained from all 73 dogs, just before the beginning of the sandfly season. PCR was found positive in 46/73 (63%) whereas, IFAT only in 9/73 (12.3%) of the dogs. The prevalence and the incidence of Leishmania infection by PCR were 61.9 and 47.1%, respectively. No association was found between the breed, age, sex, length of hair coat of the dog, urban or rural life and the presence of ample vegetation and water collections in the proximity of their living quarters, and the result of PCR. These findings clearly demonstrate that most of the dogs residing areas where leishmaniasis is endemic become infected but usually remain seronegative. Serological screening of the general canine population in these areas may result in an underestimation of the true prevalence of the infection rate.