Histological and parasitological distinctive findings in clinically-lesioned and normal-looking skin of dogs with different clinical stages of leishmaniosis.

BACKGROUND: Normal-looking skin of dogs with leishmaniosis frequently shows microscopic lesions along with the presence of Leishmania amastigotes. However, histological lesions with or without detection of amastigotes might not occur in less severe clinical cases. In addition, comparative studies between paired clinically-lesioned and normal-looking skin samples from dogs with different disease severity are lacking. The objective of this study was to compare histological and parasitological findings by Leishmania immunohistochemistry (IHC) and quantitative PCR (qPCR) on paired clinically-lesioned and normal-looking skin biopsies from 25 dogs with different clinical stages of leishmaniosis, 11 with stage I-mild disease (papular dermatitis) and 14 with stage II-III (ulcerative or exfoliative dermatitis). RESULTS: The study demonstrated microscopic lesions in 14 out of 25 (56%) samples from normal-looking skin biopsies. In those samples, perivascular to interstitial dermatitis composed by macrophages with lymphocytes and plasma cells was observed mainly in the superficial and mid-dermis. The intensity of the dermatitis was mild to moderate and always less prominent than in the clinically-lesioned skin. In normal-looking skin samples, the presence of parasites was detected by histology, IHC and qPCR in 5/25 (20%), 8/25 (32%) and 18/25 (72%), respectively. Leishmania was encountered in 11/25 (44%), 23/25 (92%) and 25/25 (100%) of clinically-lesioned skin samples by histology, IHC and qPCR, respectively. Normal-looking skin from dogs with stage I-mild disease was less frequently inflamed (P = 0.0172). Furthermore, Leishmania was more easily demonstrated by histology (P = 0.0464), IHC (P = 0.0421) or qPCR (P = 0.0068) in normal-looking skin of dogs with stage II-III-moderate to severe disease. In addition, in the latter group, there was a significantly higher parasite load studied by means of qPCR than in dogs with less severe disease (P = 0.043). Clinically-lesioned skin from dogs with stage I disease was more frequently characterised by the nodular to diffuse pattern and granuloma formation (P = 0.0166) and by a lower parasite load studied by means of qPCR (P = 0.043) compared with more diseased dogs. CONCLUSIONS: Normal-looking skin from dogs with stage I is less likely to present histological lesions as well as harbour the parasite when compared with dogs with moderate to severe leishmaniosis.

A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain.

There is limited information regarding the prevalence of many vector borne pathogens in Europe and especially in Spanish dogs. We investigated 206 sick and 260 clinically healthy dogs from three different regions in northeastern Spain for antibodies to Rickettsia conorii (Rc), Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Bartonella henselae (Bh), Bartonella vinsonii subsp. berkhoffii (Bvb), Leishmania infantum (Li) and Borrelia burgdorferi (Bb) and for antigen of Dirofilaria immitis (Di). Total prevalences were the following: Rc (56.4%), Li (30%), Ec (16.7%), Bh (16.8%), Ap (11.5%), Bvb (1.07%), Di (0.6%) and Bb (0.6%). Seroprevalences for Rc, Ec, Ap, Bh, and Bvb and Bb and Di antigens were similar among the three different study sites. The Ec seroprevalence, as determined by Snap 3DX, was statistically lower in dogs from Mallorca (0%) than Tarragona (16%) and Barcelona (5%) (P < 0.0001). Detection of Rc antibodies was associated with seroreactivity to Ec and Ap antigens (P = 0.018 and P = 0.002, respectively). IFA Ec antibodies were associated with Ap seroreactivity (P < 0.0001). There was no association between the clinical status, sex, time of the year when samples were collected, life-style or exposure to fleas or ticks and a positive test result for Ec, Bh, Bvb, or Bb antibodies or Di antigens. Li seroreactivity was associated with illness and living outdoors (P < 0.0001, P = 0.029; respectively), Rc seroreactivity with the male gender (P = 0.028) and Ap seroreactivity with living outdoors (P = 0.045). This study indicates that exposure to Rc, Li, Ec or related Ehrlichia spp., Bh and Ap or a related spp., is common whereas Di, Bb and Bvb is uncommon among dogs from the Mediterranean basin. We also provide serological data that suggests the existence of a novel Ehrlichia species on Mallorca island.