A. F. Koutinas

Directions for the diagnosis, clinical staging, treatment and prevention of canine leishmaniosis

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.

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.

Canine atopic dermatitis in Greece: clinical observations and the prevalence of positive intradermal test reactions in 91 spontaneous cases

Atopic dermatitis was diagnosed in a total of 91 dogs by combining the compatible historical evidence and clinical signs with the presence of one or more positive intradermal test reactions well correlated with the exposure to the aeroallergens and the seasonality of the clinical signs. Compared to the general hospital population Yorkshire terriers, Chinese Shar-Peis and cocker spaniels showed a strong predilection. No such predilection was found regarding the sex of the animals. The age of the dogs at the onset of the clinical signs ranged from 2 months to 8 years (median: 2.5 years). Moderate to severe pruritus, noticed in all the 91 dogs, was either localized (29/91) or generalized (64/91) and non-seasonal (43/91), seasonal (19/91) or of unknown seasonality (29/91). The most common cutaneous lesions included erythema, hyperpigmentation, hypotrichosis and crusts; their body distribution was generalized (64%) or localized (36%) with the feet as the most common site of involvement. Five dogs that had unlesional skin were significantly younger and had been pruritic for a shorter period of time compared to the majority of our study population. Otitis externa (43/91) and bacterial pyoderma (30/91) were the most common conditions associated with atopic dermatitis, while the prevalence of Malassezia dermatitis was very low (2/91). Of the other allergic skin diseases flea allergic dermatitis was the most common (29/91) followed by food hypersensitivity (2 out of the 15 dogs tested). The majority of the dogs demonstrated multiple sensitivities to the 50 aeroallergens tested, while domestic mites (77/91), and particularly Dermatophagoides farinae (64/91), were the most commonly implicated. The total number of the positive intradermal test reactions was increasing parallel to the age of the dogs but it was negatively associated with the presence of skin lesions on the carpal and tarsal joints.

Masticatory and skeletal muscle myositis in canine leishmaniasis (Leishmania infantum)

Twenty-four dogs with a parasitologically and serologically established diagnosis of leishmaniasis were studied to investigate the atrophy of the masticatory muscles which commonly occurs in this disease, and to compare the lesions in the masticatory muscles with those in the cranial tibial muscles. The 24 animals were divided into three groups of eight, group A dogs with no muscular atrophy, group B dogs with different degrees of atrophy in the masticatory and skeletal muscles, and group C dogs with similar degrees of atrophy in the masticatory and skeletal muscles. Increased activities of creatine phosphokinase and lactate dehydrogenase were recorded in only some of the dogs in groups B and C, but there were no significant differences between the mean activities in the three groups. Electromyographic changes indicating myopathy and involving both the temporalis and cranial tibial muscles, were observed in two of the dogs in group A, seven of those in group B, and in all the dogs in group C. Muscle histopathology revealed a variable degree of muscle fibre necrosis and atrophy, mononuclear infiltrates and neutrophilic vasculitis in all the dogs except two in group A. Leishmanial amastigotes were found within macrophages and myofibres in 16 of the dogs, some in each group. IgG immune complexes were detected in muscle samples, and circulating antibodies against myofibres were detected in serum samples from all the 24 dogs.

Effects of allopurinol treatment on the progression of chronic nephritis in canine leishmaniosis (Leishmania infantum)

Forty dogs with canine leishmaniosis (CL) participated in this study, which was designed to investigate the effect of allopurinol on the progression of the renal lesions associated with this disease. The animals were allocated into 5 groups. Group A dogs (n = 12) had neither proteinuria nor renal insufficiency, group B dogs (n= 10) had asymptomatic proteinuria, and group C dogs (n = 8) were proteinuric and azotemic. Two more groups, CA and CB, comprising 5 dogs each, served as controls for groups A and B, respectively. Group A, B, and C dogs received allopurinol PO (10 mg/kg q12h) for 6 months, whereas group CA and CB dogs were placebo-treated. Serum biochemistry profile, urinalysis, urine protein/creatinine ratio, and glomerular filtration rate (GFR) measurements were carried out at the beginning of the study, the 3rd month, and the 6th month, whereas renal biopsies were carried out only at the beginning and the end of the trial. Membranoproliferative glomerulonephritis was the most common cause of chronic renal failure. Mesangioproliferative and tubulointerstitial nephritis were detected even in group A and CA dogs. Allopurinol not only lowered proteinuria in group B dogs but also prevented the deterioration of GFR and improved the tubulointerstitial, but not the glomerular, lesions in both group A and group B dogs. Further, it resolved the azotemia in 5 of the 8 dogs admitted with 2nd stage chronic renal failure (group C). Consequently, treatment with allopurinol is advisable in CL cases with asymptomatic proteinuria or 1st-2nd stage chronic renal failure.

Factors affecting the frequency of ear canal and face infestation by Otodectes cynotis in the cat

Otodectes cynotis is responsible for at least 50% of canker cases diagnosed in cats world-wide. The role of Demodex cati in the pathogenesis of otitis and acne is still obscure. The aims of this study were to estimate the prevalence of O. cynoyis and D. cati infestations in clinically normal cats in northern Greece, to determine the factors that are associated with the probability and severity of infestation in the cat, and to examine the importance of these mites in the pathogenesis of feline acne. Samples from 161 cats were examined by flushing the ear canals and by taking skin scrapings of the chin and lip area. The results were combined with various factors (sex, age, living style, hair coat type and presence of pruritus, of ear discharge, of acne-like lesions) in order to carry out a risk analysis. Two separate logistic regression analyses were performed. One, on the infestation/non-infestation potential with O. cynotis and the other, on the degree of such infestation as mild-to-moderate (< or =5 mites/field) or severe (>5 mites/field). D. cati was not detected in any of the 161 cats. The prevalence of O. cynotis was estimated at 25.5% (95% confidence interval (CI) 19-32). The rate of mite infestation was higher with the presence of ear discharge (odds ratio 9, 95% CI 3.3-24.5), periaural pruritus (odds ratio 3.6, 95% CI 1.8-8) and acne-like lesions (odds ratio 3.3, 95% CI 1.2-9). Cats with mild-to-moderate degree of infestation had 18 times higher chance of exhibiting an ear discharge than those with a severe infestation. The log-odds of mild-to-moderate parasitism were linearly related to the age.

Molecular Documentation of Bartonella Infection in Dogs in Greece and Italy

ABSTRACT 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.

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.

Tick infestation of dogs in Thessaloniki, northern Greece.

Abstract. Ticks were collected from 249 dogs, admitted over a one‐year period (1996–1997) to a Veterinary Teaching Hospital and four private veterinary practices located in the county of Thessaloniki, Greece. A total of 2812 ticks (Acari: Ixodidae) were collected, of which 2511 (89.3%) were Rhipicephalus sanguineus (Latreille) adults (1070 males, 1441 females), 156 (5.5%) R. turanicus Pomerantsev adults (35 males, 121 females), 100 (3.6%) Rhipicephalus spp. nymphs and 45 (1.6%) Rhipicephalus spp. larvae. Dogs living outdoors, in rural areas and in close proximity to farm animals, were infested with higher numbers of ticks than dogs living indoors. Tick attachment sites were recorded on 237 dogs, harbouring a total of 1711 ticks. The most heavily infested sites were, in decreasing frequency, the ear pinnae, neck, interdigital skin folds, trunk, head, ventrum, extremities and the tail. Cutaneous lesions at the attachment sites were noticed in 129 dogs and then mainly in the more severely infested animals.