Poliartrite em cão por Leishmaniose - tratamento com miltefosina / Polyarthritis in a dog due to leishmaniasis - treatment with miltefosine

Background: Canine visceral leishmaniasis causes several clinical signs, such as lymphadenomegaly, exfoliative dermatitis, ulcerative skin lesions, and lameness. The most commonly reported locomotor changes are claudication, edema, arthralgia, joint stiffness, and muscle atrophy. Radiographic exam revealed cortical and medullary destruction, increase, or decrease in medullary opacity, proliferative periosteal reaction, osteolysis, collapse of joint spaces and soft tissue edema are observed. The aim of this report is to describe the clinical and radiographic evolution of a case of erosive polyarthritis associated with leishmaniasis in a dog before, during and after treatment with miltefosine. Case: A 7-month-old mixed-breed dog was attended due pain and limited mobility. In the orthopedic evaluation, joint swelling, stiffness, and increased pain sensitivity of the four limbs, as well as neck stiffness, were noted. Radiographic examination showed joint changes compatible with edema, with increased volume and radiopacity of the soft tissues adjacent to the joints. The segments of the patient's spine showed more severe bone alterations, the cervical spine being one of the most affected regions, with multiple bone proliferations throughout the vertebral body, especially in the ventral portion (spondylosis), compatible with polyarthritis due to leishmaniasis. Due to the suspicion, lymph node and spleen cytology was performed, confirming the diagnosis. Hematological examination revealed anemia, leukopenia due to lymphopenia and thrombocytopenia in addition to increased AST (79,4 U/L; reference: 6,2 - 13 U/L), creatine kinase (517,6 U/L; reference: 1,5 - 28,4 U/L), lactate dehydrogenase (688,4 IU/L; reference: 45 - 233 IU/L) and hyperproteinemia (7,34 g/dL; reference: 5,4 - 7,1 g/dL). Treatment with miltefosine, allopurinol, domperidone, prednisone, gabapentin and dipyrone was started. Reassessments were performed monthly for 3 consecutive months. Hematological examinations showed improvement, with resolution of anemia and thrombocytopenia, and a marked decrease in creatine kinase values. Thus, it is evident that the dog did not develop liver or kidney changes during treatment. During the treatment and monitoring in this period, the dog had a clinical improvement, which started to walk without pain. In addition, joint swellings were no longer present, however, there was no improvement in the radiographic evaluation of the joints. Discussion: Clinical signs of the locomotor system are compatible with those described in animals that had osteoarticular manifestations associated with leishmaniasis, such as arthralgia, edema, and joint stiffness. In the present report, treatment with miltefosine associated with allopurinol resulted in an improvement in the clinical picture, and this therapy is therefore promising in dogs with polyarthritis due to leishmaniasis. A case published in human medicine demonstrated the intra-articular absorption capacity of this drug. There is only one study to date that describes the radiographic evolution of a dog with arthritis due to leishmaniasis after treatment with miltefosine and allopurinol. In this case described, the dog reported remained with the osteoarticular lesions after treatment, although clinical improvement was observed, as in our report. The use of miltefosine and allopurinol are in accordance with stage II staging for leishmaniasis. In this study, although there was no improvement in the radiographic examinations, the treatment was effective in the remission of the animal's clinical condition.

Transmissible Venereal Tumor associated with cutaneous metastasis and leishmaniasis in a bitch

Background: The Transmissible Venereal Tumor (TVT) is a round cell neoplasia, of contagious nature, common in the canine species, which mainly affects external genitalia. Despite metastasis being uncommon, the extragenital involvement canoccur via the lymphatic or hematogenous route to regional lymph nodes, skin, subcutaneous tissue, nasal and oral mucosa,as well as the central nervous system. When the location of the tumor is genital the clinical diagnosis can be conclusive,however if it presents extragenital forms, cytological or histopathological studies are necessary for the confirmation. Thiswork describes a case of TVT with cutaneous metastasis in a female dog with leishmaniasis.Case: A 3-year-old crossbred female dog was attended at the Small Animal Medical Clinic (CMPA) of the UniversityVeterinary Hospital (HVU) of the Federal University of West Bahia (UFOB), Barra Multidisciplinary Center. This neuteredbitch dewormed, never vaccinated, rescued from the streets at the age of two and a half, had a prior history of ehrlichiosisand pyometra. The owner reported apathy, anorexia, cachexia, depression and sternal decubitus of the animal, as wellas episodes of vomiting and recurrent fever. During the physical examination were observed respiratory and heart rateswithin normality ranges, with predominance of slightly pale mucous membranes, reactive left prescapular lymph node andsubcutaneous nodular mass, not adhered to the musculature, located in the medial portion of the thirteenth left rib. Werealso evidenced onychogryphosis, opaque, shineless and brittle fur, with pruritic and lichenified exfoliative dermatitis inthe left scapular region, ear tips and snout, presence of a discreet quantity of brownish-colored vulvar discharge with aputrid odor. In the cytological examination, was verified the presence of cells with characteristics of TVT located in thegenital mucosa, developing atypical...

Transmissible Venereal Tumor associated with cutaneous metastasis and leishmaniasis in a bitch

Background: The Transmissible Venereal Tumor (TVT) is a round cell neoplasia, of contagious nature, common in the canine species, which mainly affects external genitalia. Despite metastasis being uncommon, the extragenital involvement canoccur via the lymphatic or hematogenous route to regional lymph nodes, skin, subcutaneous tissue, nasal and oral mucosa,as well as the central nervous system. When the location of the tumor is genital the clinical diagnosis can be conclusive,however if it presents extragenital forms, cytological or histopathological studies are necessary for the confirmation. Thiswork describes a case of TVT with cutaneous metastasis in a female dog with leishmaniasis.Case: A 3-year-old crossbred female dog was attended at the Small Animal Medical Clinic (CMPA) of the UniversityVeterinary Hospital (HVU) of the Federal University of West Bahia (UFOB), Barra Multidisciplinary Center. This neuteredbitch dewormed, never vaccinated, rescued from the streets at the age of two and a half, had a prior history of ehrlichiosisand pyometra. The owner reported apathy, anorexia, cachexia, depression and sternal decubitus of the animal, as wellas episodes of vomiting and recurrent fever. During the physical examination were observed respiratory and heart rateswithin normality ranges, with predominance of slightly pale mucous membranes, reactive left prescapular lymph node andsubcutaneous nodular mass, not adhered to the musculature, located in the medial portion of the thirteenth left rib. Werealso evidenced onychogryphosis, opaque, shineless and brittle fur, with pruritic and lichenified exfoliative dermatitis inthe left scapular region, ear tips and snout, presence of a discreet quantity of brownish-colored vulvar discharge with aputrid odor. In the cytological examination, was verified the presence of cells with characteristics of TVT located in thegenital mucosa, developing atypical...(AU)

Cytological, histopathological, histochemical, and immunohistochemical findings of multiple cutaneous nodules in a bovine / Achados citológicos, histopatológicos, histoquímicos e imuno-histoquímicos de múltiplas nodulações cutâneas em um bovino

This case study describes the cytological and histopathological findings of cutaneous masses in a bovine, including a peripheral nerve sheath tumor (PNST), vaccine-associated granulomatous inflammation, and eosinophilic inflammation due to parasitosis. A six-year-old undefined cow (SRD) presented with heterogeneous cutaneous lesions including multiple nodules in the left paralumbar fossa, bilaterally at the withers, and scattered along the dorsum, limbs and near the tail; some lesions were associated with ticks. Cytology of these nodules showed benign mesenchymal neoplasia (paralumbar fossa), granulomatous and pyogranulomatous inflammation (withers) and keratin (dorsum). Histopathology, in this order, confirmed PNST, post-vaccination granuloma, and eosinophilic dermatitis. A peripheral nerve sheath tumor was suspected based on the histological findings, showing a well-delineated proliferation of fusiform cells arranged in plexiform structures, which appeared red by Masson's Trichrome stain. The diagnosis was confirmed by immunohistochemistry (anti-S100 antibody). Vaccine reaction often occurs in cattle, and cytological examination is sufficient to determine the inflammatory process. Eosinophilic dermatitis is usually accompanied by perivascular inflammation and reflects the exfoliative process by the oral apparatus of the parasite.

Descrevem-se os achados citológicos e histopatológicos do tumor de bainha de nervo periférico (TBNP), da reação vacinal e da inflamação eosinofílica decorrente dapicada de carrapatoem um bovino. Uma vaca sem raça definida (SRD) de seis anos de idade foi apresentada com diferentes lesões cutâneasnodulares localizadas na fossa paralombar esquerda, bilateralmente na cernelha e dispersos no dorso, membros e próximo à cauda, por vezes associado a carrapatos. Realizou-se citologia e biópsia desses nódulos. Na citologia verificou-se neoplasia mesenquimal benigna (fossa paralombar), inflamação granulomatosa e piogranulomatosa (cernelha) e ceratina (dorso). Na histopatologia, confirmou-se que esses nódulos correspondiam, nessa ordem, a tumor de bainha de nervo periférico, granuloma vacinal e dermatite eosinofílica. O diagnóstico do TBNP foi estabelecido com base nos achados histológicos, que caracterizaram-se por uma proliferação bem delimitada de células fusiformes arranjadas em estruturas plexiformes, corados em vermelho pelo Tricômico de Masson, e confirmado por imuno-histoquímica (anticorpo anti-S100). A reação vacinal ocorre frequentemente em bovinos e o exame citológico é suficiente para determinação do processo inflamatório. Dermatite eosinofílica em geral é acompanhada de inflamação perivascular e perianexal e reflete a ação esfoliativa do

CD45+, CD68+ and E-cadherin+ Expressions in Skin Dogs Naturally Infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+, CD68+ and E-cadherin+  associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum.Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+, CD68+ and E-cadherin+, and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and bioch

CD45+, CD68+ and E-cadherin+ expressions in skin dogs naturally infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+ , CD68+ and E-cadherin+ associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum. Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+ , CD68+ and E-cadherin+ , and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and [...](AU)

CD45+, CD68+ and E-cadherin+ expressions in skin dogs naturally infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+ , CD68+ and E-cadherin+ associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum. Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+ , CD68+ and E-cadherin+ , and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and [...]

Parasite load in intact and ulcerative skin of dogs with leishmaniais

Abstract The skin is the site of inoculation of Leishmania spp. in susceptible hosts, and consequently dermatopathies, especially ulcerative dermatitis, are the main clinical signs observed. The aim of this study was to assess parasitism of the skin (intact and ulcerated) among dogs that were naturally infected by Leishmania spp., through immunohistochemical analysis. Skin fragments (intact and ulcerated) were collected from 13 dogs with positive parasitological (bone marrow aspiration and exfoliative skin) and serological examinations (ELISA S7® Biogene) forLeishmania spp. These samples were processed using the immunohistochemical technique, involving the streptavidin-peroxidase complex. Ulcerative lesions were mainly observed on the elbows (53.84%; 7/13), nostrils (15.38%; 2/13), ears (23.07%; 3/13) and wings of the ilium (7.69%; 1/13). A severe parasite load was detected in 46.15% and 76.92% of the intact and ulcerated skin samples tested, respectively. The parasite load on ulcerated skin was statistically higher than on intact skin (p = 0.0221). These results indicate that the intact and ulcerated skin may host a high parasite load of amastigote forms of Leishmania spp., which can favor the transmission of the parasite.

Resumo A pele é o local de inoculação da Leishmania spp. nos hospedeiros susceptíveis e dessa forma, as dermatopatias, principalmente as dermatites ulcerativas são os principais sinais clínicos observados. O objetivo deste estudo foi avaliar o parasitismo na pele (íntegra e ulcerada) em cães naturalmente infectados por Leishmania spp. através da técnica de imunohistoquímica. Fragmentos de pele (íntegra e ulcerada) foram coletados de 13 cães com diagnóstico parasitológico (aspirado de medula óssea e esfoliação cutânea) e sorológico positivos (ELISA S7® Biogene) paraLeishmania spp. Amostras foram processadas por imunohistoquímica pelo complexo estreptoavidina-peroxidase. As lesões ulcerativas foram observadas principalmente nas regiões do cotovelo 53,84% (7/13), narina 15,38% (2/13), orelha 23,07% (3/13) e sobre a asa do ílio 7,69% (1/13). Uma intensa carga parasitária foi detectada 46,15% e 76,92% das amostras de pele íntegra e ulcerada, respectivamente. A carga parasitária na pele ulcerada foi estatisticamente superior à pele íntegra (p = 0,0221). Esses resultados indicam que a pele intacta e ulcerada pode albergar uma intensa carga parasitária de formas amastigotas de Leishmania spp., o que pode favorecer a transmissão do parasita.

Parasite load in intact and ulcerative skin of dogs with leishmaniais / Carga parasitária em fragmentos de pele intacta e ulcerada em cães com leishmaniose

The skin is the site of inoculation of Leishmania spp. in susceptible hosts, and consequently dermatopathies, especially ulcerative dermatitis, are the main clinical signs observed. The aim of this study was to assess parasitism of the skin (intact and ulcerated) among dogs that were naturally infected by Leishmania spp., through immunohistochemical analysis. Skin fragments (intact and ulcerated) were collected from 13 dogs with positive parasitological (bone marrow aspiration and exfoliative skin) and serological examinations (ELISA S7® Biogene) forLeishmania spp. These samples were processed using the immunohistochemical technique, involving the streptavidin-peroxidase complex. Ulcerative lesions were mainly observed on the elbows (53.84%; 7/13), nostrils (15.38%; 2/13), ears (23.07%; 3/13) and wings of the ilium (7.69%; 1/13). A severe parasite load was detected in 46.15% and 76.92% of the intact and ulcerated skin samples tested, respectively. The parasite load on ulcerated skin was statistically higher than on intact skin (p = 0.0221). These results indicate that the intact and ulcerated skin may host a high parasite load of amastigote forms of Leishmania spp., which can favor the transmission of the parasite.(AU)

A pele é o local de inoculação da Leishmania spp. nos hospedeiros susceptíveis e dessa forma, as dermatopatias, principalmente as dermatites ulcerativas são os principais sinais clínicos observados. O objetivo deste estudo foi avaliar o parasitismo na pele (íntegra e ulcerada) em cães naturalmente infectados por Leishmania spp. através da técnica de imunohistoquímica. Fragmentos de pele (íntegra e ulcerada) foram coletados de 13 cães com diagnóstico parasitológico (aspirado de medula óssea e esfoliação cutânea) e sorológico positivos (ELISA S7® Biogene) paraLeishmania spp. Amostras foram processadas por imunohistoquímica pelo complexo estreptoavidina-peroxidase. As lesões ulcerativas foram observadas principalmente nas regiões do cotovelo 53,84% (7/13), narina 15,38% (2/13), orelha 23,07% (3/13) e sobre a asa do ílio 7,69% (1/13). Uma intensa carga parasitária foi detectada 46,15% e 76,92% das amostras de pele íntegra e ulcerada, respectivamente. A carga parasitária na pele ulcerada foi estatisticamente superior à pele íntegra (p = 0,0221). Esses resultados indicam que a pele intacta e ulcerada pode albergar uma intensa carga parasitária de formas amastigotas de Leishmania spp., o que pode favorecer a transmissão do parasita.(AU)

Canine visceral leishmaniasis: dermatological aspects and associated dermatoses / Leishmaniose visceral canina: aspectos dermatológicos e dermatoses associadas

Background: Visceral leishmaniasis (VL) is a zoonotic disease caused by Leishmania chagasi (syn. L. infantum). The dog is the main reservoir for this infectious agent in the urban environment. Among the various systemic signs of viscerotropic infection by L. chagasi, cutaneous lesions, including exfoliative dermatitis, cutaneous ulcers and nodules, alopecia, papular or pustular dermatitis, and onychogryphosis, are the most common in dogs. This study aimed to describe the major cutaneous lesions, evaluate the skin parasite L. chagasi by PCR, and investigate the main dermatoses associated with this zoonosis. Materials, Methods & Results: This study evaluated 50 seropositive dogs of various breeds and sizes for VL by ELISA and IFA and for the dermatological signs associated with VL. Moreover, molecular analysis of skin fragments was performed with primers 150 and 152 for the genus Leishmania, and the species was verified as L. chagasi with RV1 and RV2 primers. Deep skin scraping for mites and fungal culture analysis were performed in all dogs. Of the 50 dogs, 15 (30%) were free of systemic or cutaneous signs; however, changes in skin and annexes were observed in 35 (70%) dogs. Thirty-one dogs (62%) presented infection with dermatophytes, 26 (83.9%) with Microsporum sp., and 5 (16.1%) with Trichophyton sp.; only one dog showed parasitism by Sarcoptes scabiei. A statistically [...](AU)

Canine visceral leishmaniasis: dermatological aspects and associated dermatoses / Leishmaniose visceral canina: aspectos dermatológicos e dermatoses associadas

Background: Visceral leishmaniasis (VL) is a zoonotic disease caused by Leishmania chagasi (syn. L. infantum). The dog is the main reservoir for this infectious agent in the urban environment. Among the various systemic signs of viscerotropic infection by L. chagasi, cutaneous lesions, including exfoliative dermatitis, cutaneous ulcers and nodules, alopecia, papular or pustular dermatitis, and onychogryphosis, are the most common in dogs. This study aimed to describe the major cutaneous lesions, evaluate the skin parasite L. chagasi by PCR, and investigate the main dermatoses associated with this zoonosis. Materials, Methods & Results: This study evaluated 50 seropositive dogs of various breeds and sizes for VL by ELISA and IFA and for the dermatological signs associated with VL. Moreover, molecular analysis of skin fragments was performed with primers 150 and 152 for the genus Leishmania, and the species was verified as L. chagasi with RV1 and RV2 primers. Deep skin scraping for mites and fungal culture analysis were performed in all dogs. Of the 50 dogs, 15 (30%) were free of systemic or cutaneous signs; however, changes in skin and annexes were observed in 35 (70%) dogs. Thirty-one dogs (62%) presented infection with dermatophytes, 26 (83.9%) with Microsporum sp., and 5 (16.1%) with Trichophyton sp.; only one dog showed parasitism by Sarcoptes scabiei. A statistically [...]

PARÂMETROS INFLAMATÓRIOS E IMUNOEXPRESSÃO DE TLR-2, TLR-4, COX-2 e VEGF NA PELE DE CÃES NATURALMENTE INFECTADOS POR Leishmania infantum

A pele é a primeira barreira mecânica de proteção contra patógenos do ambiente. Na leishmaniose visceral canina (LVC), a infecção ocorre pelo rompimento da barreira da pele pelo flebotomíneo infectado que inocula o protozoário Leishmania infantum. Esses agentes infecciosos desencadeiam respostas imune e inflamatória severas no animal que podem ser avaliadas sistemicamente e no local da infecção. Na LVC, uma resposta inflamatória é desencadeada no sítio infeccioso por células sentinelas que liberam mediadores inflamatórios e citocinas e induzem ao recrutamento de células inflamatórias. Neste trabalho objetivou-se caracterizar o perfil hematológico e bioquímico e avaliar as expressões de VEGF+, COX-2+, TLR-2+ e TLR-4+ na pele de cães naturalmente infectados por L. infantum. Para tanto, cães soropositivos (n=33) foram divididos nos grupos assintomático (GA, n=10) e sintomático (GS, n=23), de acordo com a presença ou ausência de alterações dermatológicas, comparados com o grupo de animais soronegativos (GC, n=5). As principais manifestações clínicas observadas em GS foram alopecia, onicogrifose, ceratoconjuntivite e dermatites eritematosas, esfoliativas e ulcerativas. Amostras de sangue foram coletadas para avaliação dos parâmetros hematológicos e bioquímicos. Biópsias da pele foram coletadas da região escapular e da ponta de orelha, processados para histologia para avaliação do infiltrado inflamatório (ausente, leve, moderado e intenso) e imunoistoquímica com anticorpos monoclonais anti-VEGF+, anti-COX-2, anti-TLR-4 e anti-TLR-2 (ausente, leve, moderado e intenso). Os animais sintomáticos para LVC apresentaram albuminemia, leucocitose e outras alterações hematológicas compatíveis com descrições da literatura. A contagem diferencial leucocitária apontou para uma neutrofilia e linfopenia significativas do GS quando comparadas aos demais grupos (p < 0,05), estando esse resultado confirmado pela razão neutrófilo:linfócito, também significativamente aumentada. Na histologia foi evidenciado infiltrado inflamatório intenso em GS em relação ao GA e GC. As expressões de VEGF+, COX-2+ e TLR-2 estavam aumentadas na pele de animais GS (intenso) quando comparadas a GA (moderado) e GC (leve), enquanto que a marcação de TLR-4, não foi alterada entre os grupos avaliados. Estes dados indicam o envolvimento de VEGF+ e COX-2, pela ativação do TLR-2, o que pode sugerir uma relação da via de transcrição do NF-kB pelo TLR-2 e não pelo TLR-4 em resposta a infecção por L. infantum.

The skin is a mechanical barrier in the protection against environmental pathogens. In Canine Leishmaniasis (CanL), the infection occurs through phlebotomine vectors that inoculate Leishmania infantum through the skin. These infectious agents elicit severe immune and inflammatory responses in the animal that can be assessed both systemically and at the site of infection. In response to the infection, sentinel cells triggered an inflammatory response trough the release of mediators and cytokines that induce recruitment of inflammatory cells to the local of infection. The objective of this study was to characterize the hematological biochemical profile and to evaluate the expression of VEGF+, COX-2+, TLR-2+ and TLR-4+ in the skin of dogs naturally infected by L. infantum. Therefore, seropositive dogs (n = 33) were divided into asymptomatic (AG, n = 10) and symptomatic (SG, n = 23) groups, according to the presence or absence of dermatological alterations, compared to the seronegative group (CG, n = 5). The main clinical manifestations observed in SG were alopecia/hypotrichosis, onychogryphosis, keratoconjunctivitis and erythematous, exfoliative and ulcerative dermatitis. Blood samples were collected for evaluation of hematological and biochemical parameters. Skin biopsies were collected from the scapular region and the ear tip, processed for histology to evaluate the inflammatory infiltrate (absent, mild, moderate and intense) and for immunohistochemistry to evaluate the expressions of VEGF+, COX-2, TLR-4 and TLR-2 (absent, mild, moderate and intense). The symptomatic animals for LVC presented albuminemia, leukocytosis and other hematological alterations compatible with descriptions in literature. The differential blood count shows a significant neutrophilia and lymphopenia in CG when compared to the other groups (p <0.05), and this result was confirmed by the neutrophil: lymphocyte ratio, also significantly increased. In the histological evaluation, both SG and AG presented inflammatory infiltrate. The expressions of VEGF +, COX-2 + and TLR-2 were increased at skin from dogs SG (intense) when compared to AG (moderate) and CG (mild), whereas TLR-4 was not detected between groups evaluated. These data indicate the involvement of TLR-2 in CanL and suggest the NF-kB transcription factor pathway to the VEGF + and COX-2 expressions in cutaneous immune system in response to L. infantum. TLR-4 not participate of this event in dogs naturally infected by L. infantum.

EXPRESSÕES DE CD45+, CD68+ e E-CADERINA+ NA PELE DE CÃES NATURALMENTE INFECTADOS POR Leishmania infantum

A pele é composta por um sistema imune cutâneo responsável pela primeira linha de defesa aos insultos ambientais e patogênicos. Na leishmaniose canina, a infecção inicia-se nesse órgão através da picada de vetores flebotomíneos que inoculam na derme o protozoário Leishmania infantum, parasita intracelular do sistema fagocítico mononuclear. O objetivo deste trabalho foi avaliar a expressão de CD45+, CD68 + e E-caderina+ associando às células sentinelas da pele e caracterizar o padrão celular inflamatório correlacionando as lesões cutâneas de cães naturalmente infectados por L.infantum. Para tanto, cães infectados (n = 22) por L.infantum foram divididos em grupo assintomático (AD) e grupo sintomático (SD), baseados na presença ou ausência de alterações cutâneas. Cães não infectados (CD, n = 5) foram incluídos como grupo controle. Amostras de biópsias de pele foram coletadas de áreas da região escapular e do terço superior da orelha. Histologia de rotina, coloração HE e azul de toluidina, e imuno-histoquímica com anticorpos monoclonais contra CD45+, CD68 + e Ecaderina+ foram realizadas. Todo o protocolo experimental foi submetido ao Comitê de Ética em Uso de Animais (CEUA) da Universidade Estadual do Ceará (UECE) sob protocolo nº6459434/2015. Alterações cutâneas como ceratoconjuntivite, onicogrifose, liquenificação, despigmentação, alopecia, hipotricose, dermatite eritematosa, dermatite esfoliativa, dermatite ulcerativa e dermatite crostosa foram observadas no SD, destacando-se a dermatite ulcerativa e a alopecia. Além disso, no grupo SD, observou-se uma freqüência significativa de infiltração celular cutânea na derme superficial e profunda em relação à AD, composta por macrófagos, plasmócitos, linfócitos e neutrófilos. Houve maior expressão dos mastócitos em SD com infiltrado intenso e AD quando comparado aos outros animais, enquanto CD68+, CD45+ e E-caderina+ foram mais expressos apenas em SD. Nesse contexto, concluiu-se que as expressões de CD45+, CD68 + e E-caderina+ caracterizam a ativação das células imunitárias sentinelas que promovem o recrutamento do infiltrado celular, que por sua vez influencia diretamente no aparecimento das lesões cutâneas, tais como dermatite ulcerativa e alopecia. Por outro lado, não foi possível observar associação entre os mastócitos e a evolução clínica. Diante disso, esses dados podem contribuir para o acompanhamento da progressão da leishmaniose canina na pele.

The skin is composed of a cutaneous immune system responsible for the first line of defense to environmental and pathogenic insults. In canine leishmaniasis, infection begins in this organ through the bite of phlebotomine vectors that inoculate the protozoan Leishmania infantum, an intracellular parasite of the mononuclear phagocytic system. The objective of this work was to evaluate the expression of CD45+, CD68 + and E-caderina+ by associating with the sentinel cells of the skin and characterize the inflammatory cellular pattern correlating the skin lesions of dogs naturally infected by L.infantum. For this, infected dogs (n = 22) by L.infantum were divided into asymptomatic group (AD) and symptomatic group (SD), based on the presence or absence of cutaneous alterations. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies were collected from areas of the scapular region and the upper third of the ear. Routine histology, HE staining and toluidine blue, and immunohistochemistry with monoclonal antibodies against CD45+, CD68 + and Ecaderina+ were performed. The entire experimental protocol was submitted to the Committee on Ethics in Animal Use (CEUA) of the State University of Ceará (UECE) under protocol nº6459434/2015. Cutaneous alterations such as keratoconjunctivitis, onychogrifosis, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crustal dermatitis were observed in SD, including ulcerative dermatitis and alopecia. In addition, in the SD group, a significant frequency of cutaneous cell infiltration in the superficial and deep dermis was observed in relation to AD, composed of macrophages, plasma cells, lymphocytes and neutrophils. There was greater expression of mast cells in SD with intense infiltrate and AD when compared to the other animals, whereas CD45+, CD68 + and E-caderina+ were more expressed only in SD. In this context, it was concluded that in CanL the expressions of CD45+, CD68 + and E-caderina+, characterize the activation of sentinel immune cells that promote the recruitment of cellular infiltrate, which in turn directly influences the appearance of cutaneous lesions, such as ulcerative dermatitis and alopecia. On the other hand, it was not possible to observe association between the mast cells and the clinical evolution. Therefore, these data may contribute to the monitoring of the progression of canine leishmaniasis in the skin.

DESCRIÇÃO MORFOQUANTITATIVA DE LESÕES CUTÂNEAS EM CÃES COM LEISHMANIOSE VISCERAL CANINA

A Leishmaniose Visceral Canina é uma doença que acomete, dentre outros órgãos, a pele. Esse estudo teve como objetivo quantificar as alterações morfológicas e histológicas presentes nas várias apresentações cutâneas de cães naturalmente infectados. Para isso foram avaliados 31 fragmentos de pele de 20 animais com dermatite esfoliativa seca, alopecia, hipotricose, crosta, nódulo, úlcera, eritema, despigmentação, hiperpigmentação e/ou hiperqueratose. Foram determinados o padrão microscópico inflamatório e não inflamatório de cada lesão e a histomorfometria por meio da detecção e diferenciação dos colágenos do tipo I e III. As regiões acometidas foram cabeça (40,35%), membros (21,05%), tórax (15,8%), difusa (8,77%), abdômen (5,26%), pelve (5,26%) e cervical (3,51%), sendo a dermatite crostosa e a hipotricose as macroleões mais encontradas. Houve o predomínio de infiltrado histiocítico, sendo a inflamação visualizada principalmente em região peri/intra-anexo. A forma amastigota foi visualizada em 48,38% dos casos. Colagenólise, crostas e hiperqueratose ortoqueratótica foram as lesões não inflamatórias mais encontradas. Houve predominância do colágeno maduro (60,31% + 2,90) em relação ao imaturo (8,72% + 1,187) em região de derme e esses dados foram estatisticamente significativos. Houve um discreto aumento de colágeno do tipo III e diminuição do colágeno do tipo I, ao se comparar amostras com e sem presença de amastigotas. Concluiu-se que a presença do infiltrado inflamatório em cães com Leishmaniose Visceral Canina no Distrito Federal, contribuiu para as mudanças ocorridas na matriz extracelular.

The Canine Visceral Leishmaniasis is a disease that affects, among other organs, the skin. This study aimed to quantify the morphological and histological changes present in various skin samples from naturally infected dogs. To achieve the proposed objective were evaluated 31 skin flakes of 20 animals with dry exfoliative dermatitis, alopecia, hypotrichosis, crust, nodules, ulcers, erythema, depigmentation, hyperpigmentation and/or hyperkeratosis. Inflammatory and non-inflammatory microscopic pattern of each lesion, the measurement of inflammatory intensity, parasitological and collagenolysis, and histomorphometry through the detection and differentiation of type I and III collagens were performed. The regions affected were head (40.35%), members (21.05%), chest (15.8%), diffuse (8.77%), abdomen (5.26%), pelvis (5.26%) and cervical (3.51%), the crusted dermatitis and hypotrichosis large lesions were the most frequent injuries. There was a predominance of histiocytic infiltrate and most of the fragments showed severe inflammatory intensity, with inflammation viewed mainly in peri/intra-annexed region. The amastigotes were seen in 48.38% of cases. Collagenolysis, crusts and orthokeratotic hyperkeratosis were the most frequent non-inflammatory lesions. There was a predominance of mature collagen (60.31% + 2.906) compared to immature (8,720 + 1,187%) in the dermis region and these data were statistically significant. There was a slight increase in collagen type III and decreased collagen type I, when comparing samples with and without the presence of amastigotes. It was concluded that the presence of inflammatory infiltrate in dogs with CVL in DF contributed to the changes in the extracellular matrix.

Histopathological pattern of exfoliative and ulcerative skin lesions in dogs with leishmaniasis / Padrões histopatológicos das lesões descamativas e ulcerativas da pele em cães com leishmaniose

Canine visceral leishmaniasis is an endemic infectious disease in some São Paulo state regions and even though it is a systemic disease, in the dog, the main clinical sign is dermatological. Thirty dogs with positive results in serology and parasitological exams for leishmaniasis from Araçatuba city were evaluated. They just showed dermatological signs and were divided in two groups, one with exfoliative (n=1 ) and other with ulcerative (n=1 ) lesions. Histopathological patterns in the group of exfoliative lesions were: periadnexial dermatitis (5/15, 33,3%), superficial perivascular dermatitis (1/15, 6,6%), nodular dermatitis (1/1 , , %) and mixed dermatitis (8/1 , , %), including intersticial/periadnexial dermatitis (1/8, 12,5%), lichenoid/perivascular superficial and deep dermatitis (1/8, 12,5%), perivascular superficial and deep/periadnexial dermatitis (1/8, 12,5%) and superficial perivascular/perianexial dermatitis ( /8, , %). In the group of ulcerative lesions, the histopathological patterns were: perivascular superficial and deep dermatitis (5/15, 33,3%), diffuse dermatitis (3/15, 20%), periadnexial dermatitis ( /1 , 1 , %), nodular dermatitis (1/1 , , %) and mixed dermatitis ( /1 , , %), including intersticial/ perivascular superficial and deep dermatitis (1/4, 25%), nodular/periadnexial dermatitis (1/4, 25%), fibrosing/perianexial dermatitis (1/4, 25%) and perivascular

A leishmaniose visceral canina é uma doença infecciosa crônica considerada endêmica em algumas regiões do estado de São Paulo e, apesar de ser uma enfermidade sistêmica, no cão, a maioria dos sinais clínicos são dermatológicos. Foram avaliados trinta cães com diagnóstico de leishmaniose, da região de Araçatuba, pelos exames parasitológico e sorológico. Os cães apresentando apenas sinais dermatológicos foram divididos em dois grupos: um grupo com descamação e outro com ulceração. Os padrões histopatológicos da pele encontrados nos cães com lesões descamativas (n=15) foram de dermatite perianexial (5/15, 33,3%), dermatite perivascular superficial (1/15, 6,6%), dermatite nodular (1/15, 6,6%), e dentre as dermatites mistas (8/15, 53,3%), dermatite intersticial/perianexial (1/8, 12,5%), dermatite liquenóide/perivascular superficial e profunda (1/8, 12,5%), dermatite perivascular superficial e profunda/perianexial (1/8, 12,5%) e dermatite perivascular superficial/perianexial (5/8, 62,5%). Os cães com lesões ulcerativas (n=15) apresentaram padrões histopatológicos de dermatite perivascular superficial e profunda (5/15, 33,3%), dermatite difusa (3/15, 20%), dermatite perianexial (2/15, 13,3%), dermatite nodular (1/15, 6,6%), e dentre as dermatites mistas (4/15, 26,6%), dermatite intersticial/perivascular superficial e profunda (1/4, 25%), dermatite nodular/perianexial (1/4, 25%), derma

Histopathological pattern of exfoliative and ulcerative skin lesions in dogs with leishmaniasis / Padrões histopatológicos das lesões descamativas e ulcerativas da pele em cães com leishmaniose

Canine visceral leishmaniasis is an endemic infectious disease in some São Paulo state regions and even though it is a systemic disease, in the dog, the main clinical sign is dermatological. Thirty dogs with positive results in serology and parasitological exams for leishmaniasis from Araçatuba city were evaluated. They just showed dermatological signs and were divided in two groups, one with exfoliative (n=1 ) and other with ulcerative (n=1 ) lesions. Histopathological patterns in the group of exfoliative lesions were: periadnexial dermatitis (5/15, 33,3%), superficial perivascular dermatitis (1/15, 6,6%), nodular dermatitis (1/1 , , %) and mixed dermatitis (8/1 , , %), including intersticial/periadnexial dermatitis (1/8, 12,5%), lichenoid/perivascular superficial and deep dermatitis (1/8, 12,5%), perivascular superficial and deep/periadnexial dermatitis (1/8, 12,5%) and superficial perivascular/perianexial dermatitis ( /8, , %). In the group of ulcerative lesions, the histopathological patterns were: perivascular superficial and deep dermatitis (5/15, 33,3%), diffuse dermatitis (3/15, 20%), periadnexial dermatitis ( /1 , 1 , %), nodular dermatitis (1/1 , , %) and mixed dermatitis ( /1 , , %), including intersticial/ perivascular superficial and deep dermatitis (1/4, 25%), nodular/periadnexial dermatitis (1/4, 25%), fibrosing/perianexial dermatitis (1/4, 25%) and perivascular

A leishmaniose visceral canina é uma doença infecciosa crônica considerada endêmica em algumas regiões do estado de São Paulo e, apesar de ser uma enfermidade sistêmica, no cão, a maioria dos sinais clínicos são dermatológicos. Foram avaliados trinta cães com diagnóstico de leishmaniose, da região de Araçatuba, pelos exames parasitológico e sorológico. Os cães apresentando apenas sinais dermatológicos foram divididos em dois grupos: um grupo com descamação e outro com ulceração. Os padrões histopatológicos da pele encontrados nos cães com lesões descamativas (n=15) foram de dermatite perianexial (5/15, 33,3%), dermatite perivascular superficial (1/15, 6,6%), dermatite nodular (1/15, 6,6%), e dentre as dermatites mistas (8/15, 53,3%), dermatite intersticial/perianexial (1/8, 12,5%), dermatite liquenóide/perivascular superficial e profunda (1/8, 12,5%), dermatite perivascular superficial e profunda/perianexial (1/8, 12,5%) e dermatite perivascular superficial/perianexial (5/8, 62,5%). Os cães com lesões ulcerativas (n=15) apresentaram padrões histopatológicos de dermatite perivascular superficial e profunda (5/15, 33,3%), dermatite difusa (3/15, 20%), dermatite perianexial (2/15, 13,3%), dermatite nodular (1/15, 6,6%), e dentre as dermatites mistas (4/15, 26,6%), dermatite intersticial/perivascular superficial e profunda (1/4, 25%), dermatite nodular/perianexial (1/4, 25%), derma

Expression of CD3 and CD79a cell markers in exfoliative and ulcerative skin lesions in dogs with leishmaniasis / Imunomarcação de células CD3+ e CD79a+ nas lesões descamativas e ulcerativas de cães com leishmaniose

Canine visceral leishmaniasis is an endemic infectious disease in some regions from São Paulo state and even though it is a systemic disease, in the dog, the main clinical sign is dermatological. Thirty dogs with positive results in serology (IFI) and parasitological exams (fine needle aspiration) for leishmaniasis from Araçatuba city were evaluated. They only showed dermatological signs and were divided in two groups, one with exfoliative (n=15) and other with ulcerative (n=15) lesions. There was significant statistical difference between CD3 and CD79a population in the same dermatological pattern, and with CD79a population between exfoliative and ulcerative patterns (p 0,05).

A leishmaniose visceral canina é uma doença infecciosa crônica considerada endêmica em algumas regiões do estado de São Paulo e, apesar de ser uma enfermidade sistêmica, no cão, a maioria dos sinais clínicos são dermatológicos. Foram avaliados trinta cães com diagnóstico de leishmaniose, da região de Araçatuba, pelos exames parasitológico (punção aspirativa por agulha fina de linfonodos) e sorologia (IFI). Os cães apresentando unicamente sinais dermatológicos formam divididos em dois grupos: um grupo com descamação e outro com ulceração. Houve diferença estatística entre a população de células CD3+ e CD79a+ dentro do mesmo padrão dermatológico, o mesmo ocorreu com a população de CD79a+ entre os padrões descamativo e ulcerativo (p 0,05 ).

Expression of CD3 and CD79a cell markers in exfoliative and ulcerative skin lesions in dogs with leishmaniasis / Imunomarcação de células CD3+ e CD79a+ nas lesões descamativas e ulcerativas de cães com leishmaniose

Canine visceral leishmaniasis is an endemic infectious disease in some regions from São Paulo state and even though it is a systemic disease, in the dog, the main clinical sign is dermatological. Thirty dogs with positive results in serology (IFI) and parasitological exams (fine needle aspiration) for leishmaniasis from Araçatuba city were evaluated. They only showed dermatological signs and were divided in two groups, one with exfoliative (n=15) and other with ulcerative (n=15) lesions. There was significant statistical difference between CD3 and CD79a population in the same dermatological pattern, and with CD79a population between exfoliative and ulcerative patterns (p 0,05).

A leishmaniose visceral canina é uma doença infecciosa crônica considerada endêmica em algumas regiões do estado de São Paulo e, apesar de ser uma enfermidade sistêmica, no cão, a maioria dos sinais clínicos são dermatológicos. Foram avaliados trinta cães com diagnóstico de leishmaniose, da região de Araçatuba, pelos exames parasitológico (punção aspirativa por agulha fina de linfonodos) e sorologia (IFI). Os cães apresentando unicamente sinais dermatológicos formam divididos em dois grupos: um grupo com descamação e outro com ulceração. Houve diferença estatística entre a população de células CD3+ e CD79a+ dentro do mesmo padrão dermatológico, o mesmo ocorreu com a população de CD79a+ entre os padrões descamativo e ulcerativo (p 0,05 ).

CD45+, CD68+ and E-cadherin+ Expressions in Skin Dogs Naturally Infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+, CD68+ and E-cadherin+  associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum.Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+, CD68+ and E-cadherin+, and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and bioch

CD45+, CD68+ and E-cadherin+ Expressions in Skin Dogs Naturally Infected by Leishmania infantum

Background: In canine leishmaniasis (CanL), infection occurs through phlebotomine vectors that inoculate the protozoan Leishmania infantum into the skin that infected macrophages and activated dendritic cells (CD). Dogs with CanL present variable clinical manifestations, being common the presence of cutaneous lesions. The aim of this study was to evaluate the expression of CD45+, CD68+ and E-cadherin+  associating the skin sentinels cells and to compare the clinical-dermatological manifestations in the skin of dogs naturally infected by L. infantum.Materials, Methods & Results: Dogs infected (n = 22) by L. infantum were divided into asymptomatic group (AD, n = 9), and symptomatic group (SD, n = 13), according criteria based on the presence or absence of skin changes. Dogs non-infected (CD, n = 5) were included as control group. Samples of skin biopsies collected from scapular region were processed by routine histology and labeled by immunohistochemistry with monoclonal antibodies against CD45+, CD68+ and E-cadherin+, and were described as none, mild, moderate and intense. SD presented keratoconjunctivitis, onychogryphose, lichenification, depigmentation, alopecia, hypotrichosis, erythematous dermatitis, exfoliative dermatitis, ulcerative dermatitis and crusted dermatitis, and the frequency these alterations was expressed as percentage. The results of hematological and bioch