TÉCNICA DE MASSAGEM RELAXANTE COMO FERRAMENTA PARA MELHORAR A RELAÇÃO HUMANO-ANIMAL E OS PARÂMETROS DE BEM-ESTAR ANIMAL

The aversive management of farmed species may present serious consequences on productivity, reproduction, and animal welfare. Thus, studies that aim at a good human-animal relationship (HAR) and apply gentle touch, as an attempt to make this management more beneficial for humans and animals are required. In this sense, massage with bare hands is a new proposal to improve HAR with domestic ruminants. Therefore, the objectives of this review are: (1) understand the importance of a good relationship between humans and domestic ruminants, including the interaction of touch; (2) describe the impact of massage on various parameters related to physiology and emotional and behavioral states in humans and animals, and (3) propose the use of the Stroking technique (relaxing massage) in domestic ruminants as a mean to encourage producers and caretakers to use massage as an alternative method to improve HAR. Different studies and protocols have shown the importance of touch in promoting a good HAR, in addition to the various impacts of massage on humans and animals. By demonstrating that goats massaged with the Stroking technique showed positive behaviors like licking the masseur, the importance of encouraging the study of the modulation of other topics such as physiological, zootechnical, and psychological parameters in massaged domestic ruminants is highlighted. Thus, the application of relaxing massage, Stroking, as a new tool for improving human-animal interaction with domestic ruminants is a proposal that aims to create a closer relationship with animals, possibly influencing the way producers and caregivers perceive these sentient beings.O manejo aversivo de espécies de produção pode ter graves consequências na produtividade, reprodução e bem-estar animal, sendo necessários estudos que visam um bom relacionamento humano-animal (RHA), por meio do toque suave, como uma tentativa de tornar este manejo mais benéfico para seres humanos e animais. Nesse sentido, a massagem feita com as mãos constitui uma nova proposta para melhorar o RHA envolvendo ruminantes domésticos. Sendo assim, os objetivos desta revisão são: (1) compreender a importância de um bom relacionamento entre humanos e ruminantes, incluindo a interação com toque; (2) descrever o impacto da massagem sobre diversos parâmetros relacionados à fisiologia, aos estados emocionais e aos comportamentais em humanos e animais; e (3) propor o uso da técnica Stroking em ruminantes domésticos, como forma de incentivar produtores e tratadores a utilizarem a massagem como um método alternativo para melhorar o RHA. Sabe-se que diferentes estudos e protocolos mostraram a importância do toque na promoção de um bom RHA, além dos diversos impactos da massage. Ainda, ao observar que cabras massageadas com a técnica Stroking apresentaram comportamentos positivos, como lambidas na massagista, constata-se que o incentivo ao estudo da modulação de outros parâmetros, como fisiológicos, zootécnicos e psicológicos, em ruminantes domésticos massageados, é necessário. Assim, a aplicação da massagem relaxante, como uma nova ferramenta para a melhora da interação humano-animal envolvendo ruminantes domésticos, é uma proposta que visa criar uma maior proximidade com os animais, possivelmente influenciando a forma como produtores e cuidadores percebem esses seres sencientes

Successful Treatment and Management of Canine Ehrlichiosis-Leishmaniosis-Heartworm Comorbidity

 Background: Canine vector borne diseases (CVBD) are common in tropical countries where the climate favors arthropods abundance. Comorbidity with one or more CVBD are reported as clinical complication and worsen prognostic. Canine visceral leishmaniosis (CanL) is an endemic zoonotic disease in Brazil caused by Leishmania infantum, with several restrictions to canine treatment and suggestion of reservoirs euthanasia for disease control. Heart worm (HW) is a helminthic disease caused by Dirofilaria immitis infection in dogs. It is a chronic heart disease, which can lead to death by congestive heart failure. Canine ehrlichiosis (CE) is caused by Ehrlichia canis bacterial infection with a zoonotic potential and fatal to dogs in acute and chronic presentations. Exposed the above, this study aims to describe a successful treatment and management of a dog with CanL, CE, and HW comorbidity. Case: A 3-year-old male uncastrated black Labrador dog, weighing 35 kg, was admitted to the veterinary clinic due to immunochromatographic CanL positive test performed by municipal zoonosis control center active surveillance in August 2014. Clinical exam showed a mild shedding, intermittent eye white/yellow discharge and popliteal lymph nodes enlargement. After positive for CanL, veterinary requested more laboratorial exams. IFAT and ELISA were positive for CanL, blood smear showed presence of microfilaria, and bone marrow cytology showed Ehrlichia spp. morulae and microfilaria. Initial treatment prescribed was oral doxycycline, omeprazole, ranitidine, and domperidone for 30 days, and allopurinol and ivermectin until further recommendation. Additionally, repellent collar, repellent spray and vitamin supplement was indicated. After first month, marbofloxacin for 30 days and three doses of immune stimulant protocol were administrated. After three months of treatment, dog still positive for heartworm, ehrlichiosis, and CanL. Doxycycline protocol was repeated. Dog became consistently negative for all pathogens one year later with persistent thrombocytopenia but without clinical signs, ergo allopurinol and ivermectin were discontinued. After four years of follow up, the animal had an acute pancreatitis and died, with unremarkable total blood count and negative for all pathogens. Discussion: CVBD coinfections are commonly reported as worsen prognostic in endemic regions. The pathogens reported here share a host immunomodulation competence. L. infantum and Ehrlichia spp.downregulates Th1 response, whereas D. immitis increase as Th2 profile. The therapeutic protocol was iniciated by staging CanL. Since the patient had clinical signs, allopurinol was prescribed as a well-established drug for CanL. Marbofloxacin was added due to its high safety drug in clinical improvement of infected dogs with and without renal disease and in vitro effectiveness against L. infantum. Domperidone was used to promote Th1 cytokine profile as INF-γ, IL-2, IL-12, and TNF-α. We used an immunostimulant protocol to favor polarization to the Th1 profile comprised by 30 days of domperidone protocol followed by a vaccine and an immunomodulator. Doxycycline was used successfully for Ehrlichia spp. and HE clearance after two treatment courses and one year of ivermectin every 15 days. The animal presented intermittent coughing episodes on the first treatment course, but no medical intervention was needed besides exercise restriction. Our report shows the successful management of one dog with CanL, CE and HE comorbidity. This success was possible due to early detection and good therapeutic choice.Keywords: canine visceral leishmaniosis, coinfection, Dirofilaria immitis, Erhlichia canis, Leishmania infantum, treatment

Relationship between PD-L1 Expression and Tumor-Infiltrating Lymphocytes in Canine Mammary Tumor

Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in canine mammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as a shield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study, it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in canine mammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors were grouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using two thresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromal area. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs were significantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4) were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n = 9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associated with involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010) overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1– (P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymph node involvement (P = 0.004), stromal TILs (P = 0.016), and PD-L1+/TILs-High vs. PD-L1–/TILs-Low (P = 0.010). Multivariate analysis revealed that group of tumors with grade II-III was independent and negative prognostic factors for OS.Discussion: In this study, PD-L1 was differently expressed according to the histologic subtypes of TMC. Currently, has been showed the presence of PD-L1 in several canine cancer. Nevertheless, only a few studies have described PD-L1 protein expression in dog tumors and showed PD-L1 was constitutively expressed on canine tumor cell lines, although the levels of basal expression were very variable. This expression can be modulated by IFN-γ exposure. In the present study, it was found a strong PD-L1 expression on TILs. The increase in PD-L1 cell surface expression by tumor cells can lead to decreased T-cell proliferation and increased apoptosis. In human breast cancer (BC) the PD-L1 expression was expressed in TILs and tumor epithelium. It has been reported the association of stromal TILs and PD-L1 expression with aggressive types and stages of BC. In this study, it was detected PD-L1 expression in malignant epithelium in all lymph node metastasis. PD-L1 overexpression was significantly associated with a series of clinicopathological parameters. It was demonstrated that PD-L1+/TILs-High had higher risk of overall survival (OS) than another group of interaction. High PD-L1 expression may be a prognostic indicator for reduced OS, while tumor PD-L1+ was associated with poorer disease-free survival. The presence of TILs has shown to be potentially predictive and a prognostic factor in BC subtypes. In CMT, it has been reported that a high proportion of TILs was correlated to several malignancy characteristics. In relation to PD-L1, further research is necessary to clarify this immune checkpoint as a potential therapeutic target and its application in clinical practice in CMT

Avaliação dos ungüentos à base de extratos hexânico ou etanólico das folhas de Momordica Charantia L. sobre as lesões cutâneas experimentais em coelhos

Entre as inúmeras plantas empregadas na medicina tradicional destaca-se a Momordica charantia, que vem ganhando notoriedade científica por suas diversas propriedades biológicas e farmacológicas. O objetivo deste trabalho foi investigar a regeneração de lesões cutâneas em coelhos, tratadas diariamente com ungüento contendo os extratos hexânico (EH) e etanólico (EE) das folhas de M.charantia nas concentrações de 10 e 20%, comparadas com o controle (base do ungüento). Lesões cutâneas de cerca de 5mm foram induzidas experimentalmente por um punch, tendo os coelhos sido anestesiados previamente e após 4 (D4) e 14 (D14) dias de tratamento, as lesões foram removidas cirurgicamente e encaminhadas para análise histopatológica. O processo de reparo cutâneo foi avaliado nestes intervalos de tempo através da observação macroscópica, mensuração da área de retração da ferida e avaliação microscópica através da contagem do número de células mononucleares e fibroblastos e dos vasos sanguíneos. O ungüento contendo o EH (10 e 20%) demonstrou ser o mais eficaz por acelerar (

Mandibulectomy in feline with bacterial fibrosing osteomyelitis

Background: Fibrosing osteomyelitis is a chronic inflammatory process caused by infectious agents that lead to the destruction and replacement of bone tissue by fibroblasts. The diagnosis is based especially on histopathological and bacterial culture. In cases where extensive and irreversible injuries are observed, surgical treatment may be indicated. The objective of this work is to report the clinical, radiographic, histopathological, and microbiological aspects of a cat presenting fibrosing osteomyelitis.Case: A 10-year-old male feline, no defined breed, weighing 3.9 kg was referred to one Private Veterinary Clinic of Fortaleza, CE, Brazil with a history of left mandibular enlargement, presenting dysphagia, sialorrhea, an increase in firm consistency along of the left mandibular body, temporomandibular arthralgia and decreased joint motion range. On cranium radiograph, signs of proliferative osteopathy of irregular contours were observed in the branch and body of the left mandible and extending to the rostral region of the right mandible, suggesting a neoplastic process. After anesthesia, for better assessment of the oral cavity, a sample was collected by incisional biopsy, however, the histopathological result was nonspecific. In view of the inconclusive condition, it was decided to perform left hemimandibulectomy combined with right partial mandibulectomy. Tissue samples were obtained and sent for microbiological and histopathological analyses. The last test revealed an inflammatory reaction consisting of neutrophils and plasma cells, associated with a large amount of fibrous connective tissue, multifocal bacterial aggregates, necrosis and bone resorption. Based on the findings, the diagnosis of chronic bacterial osteomyelitis was concluded. The microbiological culture demonstrated the growth of the bacterium Pseudomonas aeruginosa, with sensitivity to cefovecin, which was administered to the treatment in doses of 8 mg/kg/SC, every 15 days, for 60 days. After 10 days of the surgery, the patient was able to start the process of swallowing and seizing pasty food.Discussion: Although radiography is a diagnostic method of choice to assess the extent of bone involvement, it hardly allows the distinction between neoplastic processes and osteomyelitis. Thus, in addition to the cranium radiographic examination, that revealed signs of proliferative osteopathy with irregular contours in branch and body topography of the left mandible were required biopsy and bacterial culture. After mandibulectomy, difficulty in retracting the tongue and anorexia were observed in the first days. Despite that, from the 12th day on, there was an improvement in food seizure, with the return to voluntary feeding. Mandibulectomy is indicated in cases where extensive and irreversible lesions are observed, although excision of the caudal mandible to the third or fourth premolar tooth is not recommended, because it compromises the sublingual musculature, with fall of the tongue and loss of function apprehension. In the present case, the caudal portion of the right mandibular branch was maintained, which facilitated the return of spontaneous feeding. The tissue removed was necessary to perform a microbiological culture with antibiogram, essential to determine the possible etiologic agent and choosing antimicrobial drugs. For the treatment of P. aeruginosa infection, cefovecin was prescribed due to the dosage and route of administration that collaborate with the mandibular surgery, once that the oral treatment is more difficult. Post-surgical complications related to mandibular resection, such as anorexia and difficulty in grasping food, are common. However, in the present case, the patient had an adequate reestablish after 15 days. Therefore, hemimandibulectomy is effective in treating fibrosing osteomyelitis, with maintenance of the patient's ingestive function.

Leukocytes and Albumin in Canine Leishmaniasis

Background: Canine Leishmaniasis (CanL) is a multisystemic and chronic inflammatory disease characterized by nonspecific clinical manifestations. In CanL, inflammatory cells and chemical mediators released in response to the parasite play a role in disease development and progression. Alterations on hematological parameters have been documented in CanL. These changes can also be assessed in relation to systemic inflammation caused by this disease. The circulating leukocyte counting, such as neutrophils, as well as the albumin level, are considered direct indicators of an inflammatory host environment. Several studies point to the use of biomarkers on the assistance in diagnosis and prognosis of several canine pathologies. The present study investigated the Neutrophils to Lymphocyte Ratio (NLR), Albumin to Globulin Ratio (AGR), and Neutrophils to Albumin Ratio (NAR) on systemic inflammatory response induced by Canine Leishmaniasis (CanL).Materials, Methods & Results: For this purpose, adult dogs with confirmed diagnosis to CanL were divided into symptomatic (SD, n = 33) and asymptomatic (AD, n = 20) dogs for L. infantum and control dogs (CD, n = 20). Routine hematological and biochemical parameters were determined in blood samples using a veterinary automatic hematology and biochemical analyzers. Asymptomatic dogs (AD) had a higher number of white blood cells and neutrophils (16.48 ± 4.93; 13.41 ± 3.60, respectively) in relation to symptomatic dogs (SD) (13.54 ± 5.13; 10.42± 3.69, respectively) (P = 0.015 and P < 0.0001, respectively). Neutrophils to Lymphocyte Ratio (NLR) was higher in dogs with leishmaniasis (9.45 ± 3.76) than in healthy dogs (3.39 ± 1.19) (P < 0.0001). Serum total proteins (STP) and globulins increased in CanL, while albumin and AGR decreased in CanL, when compared to CD and references values to canine species. Neutrophils to Albumin Ratio (NAR) was higher in AD and SD (5.02 ± 1.14; 4.79 ± 1.07, respectively) when compared to CD (2.36 ± 0.55) (P < 0.0001). Discussion: As reported in scientific researches, dogs with Leishmaniasis present alterations in circulating cell counts. Based on these data, we decided to expand this information using the NLR as a parameter in an attempt to better clarify the changes in these cells in CanL. We observed that NLR was increased on CanL in relation to healthy dogs, which could be a consequence of relative neutrophilia rather than lymphopenia. Neutrophils to Lymphocyte Ratio (NLR) is a biomarker that conveys information about inflammatory conditions. An elevated NLR can reflect an upregulated innate immune response, since neutrophils are effector cells of innate immunity and are involved in several acute and chronic inflammatory processes. Albumin is an acute phase protein that is considered an immune-inflammatory biomarker, which can be found reduced systemically in progressive inflammatory response. Serum total proteins (STP) and globulins were increased in CanL. These data are already well documented in CanL, which serum globulins are mainly associated with the increase of acute phase proteins, cytokines, and increase of specific antibodies to Leishmaniainfantum. Our results showed neutrophilia with hypoalbuminemia in CanL. So, in an attempt to assess the relationship of these two available markers, we used NAR calculation in order to evaluate the changes induced by CanL. In this study NAR was higher in CanL when compared to control dogs. Thus, our data indicate that NLR and NAR could be used as biomarkers in veterinary medical clinics in order to assess inflammatory profile in CanL, mainly in asymptomatic dogs. These parameters obtained from routine blood tests might be useful as cost-effective, easily accessible and helpful markers in order to distinguish the inflammatory response intensity in CanL

Analysis of Systemic and Cutaneous Inflammatory Immune Response in Canine Atopic Dermatitis

Background: Canine atopic dermatitis (CAD) is a chronic and inflammatory disease present in veterinary dermatological practice. The inflammation in CAD is triggered by environmental allergens and skin microorganisms, which are responsible for the worsening of cutaneous lesions. This continuous activation of skin inflammatory process can induce the production of free radicals that also contribute to cellular damage and ultimately leads to changes in blood parameters in dogs with CAD. Although there are reports of inflammatory parameters in CAD, there are a lack of studies correlating skin lesions, blood leukocytes and oxidative stress. Based on that,this study aimed to evaluate the integumentary and systemic inflammatory response in dogs with atopic dermatitis. Materials, Methods & Results: Dogs with confirmed diagnosis of canine atopic dermatitis (n = 10) were divided in two groups according to CADESI-IV: AI, with CADESI between 0-10, AII, with CADESI between 10-34, and control group (n = 5). Blood-biochemical and histological analysis were performed to access systemic and cutaneous inflammatory response. AII group tended to higher neutrophil and eosinophil counts, as well as neutrophil/lymphocyte ratio (NLR) when compared to AI. The albumin was lower in AII compared to AI and control (P < 0.05), while total bilirubin and malondialdehyde (MDA) did not differ between groups. NLR (r = 0.64 and P = 0.04) and MDA (r = 0.54 and P = 0.1) were positively correlated with CADESI, while albumin was negatively correlated with CADESI (r = -0.79 and P = 0.005). Histopathological analysis revealed a larger number of neutrophils, macrophages and mast cells in AI and AII than in control group (P < 0.05). Discussion: In this study it was possible to evaluate the systemic and cutaneous leukocyte dynamics in CAD. Skin inflammation induces the production of chemotactic molecules contribute to neutrophil outflow from blood vessel toward the affected tissue, which can be visualized as perivascular inflammatory infiltrate and exocytosis. At systemic level, there was a tendency to increase in total leukocytes and circulating neutrophil count in group AII, as well as in neutrophil/lymphocyte ratio (NLR), when compared to AI. The NLR is a widely available and inexpensive laboratory biomarker that quantifies systemic inflammation, being used in human medicine to evaluate prognosis in different types of cancer. In our study, dogs in AII group showed an increased NLR compared to AI and control, which demonstrates the influence of skin injury in systemic parameters. Furthermore, AII group is composed of dogs with greater lesion state, which reflects in higher NLR values. Since this disease is known by its chronicity and may remain stable for years, NLR may be a novel biomarker to evaluate acute exacerbation in CAD and could potentially explain why some patients have longer crisis duration and frequent flares. The maintenance of the inflammatory state also induces the production of oxidizing substances, which possibly exceed the total antioxidant capacity, generating a situation of oxidative stress, which can result in damage to membrane lipids and release of their products. MDA is reliable and is the most commonly used marker of the overall lipid peroxidation level and the presence of oxidative stress. This result may be related with the antioxidant system, such as albumin and bilirubin, which was able to promote an efficient control of oxidant substances. In conclusion, the presented data demonstrated an inflammatory process progression as well as introduced NLR as a potential marker of disease exacerbation in CAD

Ocorrência de baços acessórios em boto-cinza (Sotalia guianensis) - aspectos histológicos

A presença de baços acessórios tem sido relatada em seres humanos e diversas espécies de animais. Nos cetáceos, o baço é um órgão de estrutura histológica similar ao de outros mamíferos, podendo estar acompanhado, em algumas espécies, por um ou mais baços acessórios. No presente trabalho, foram avaliadas amostras de baços acessórios provenientes de um botocinza, Sotalia guianensis, encalhado no litoral do Ceará. A análise histológica das lâminas revelou a presença de estruturas similares ao órgão principal, sendo observada, no entanto, hipoplasia linfocitária. Existem poucos estudos sobre os baços acessórios em cetáceos, tendo sido descritos na maioria das vezes como achados secundários de necropsias. Desta forma, a descrição da estrutura histológica de amostras obtidas desses órgãos pode contribuir para a elucidação da sua função em organismos animais

CD4+, CD8+, FoxP3+ and HSP60+ Expressions in Cellular Infiltrate of Canine Mammary Carcinoma in Mixed Tumor

Background: Cancer is a complex process that receive many influences of the tumor microenvironment. The participation of immune system cells and proteins in tumor microenvironment is not yet completely understood. Thus, the aim of this study was to evaluate the infiltrate cellular, subpopulations of T-lymphocytes and HSP60 of canine mammary carcinoma in mixed tumor (CMCMT).Materials, Methods & Results: Female dogs (n = 20) were selected after Canine mammary tumor (CMT) diagnosis and data were achieved throughout clinical-pathological information. Clinical staging was evaluated and tumor biopsies were processed by histology and cellular infiltrate was performed according criteria and grade. Survival curve were generated by Kaplan-Meier and the lymphocytic infiltrate were compared by Log-Rank followed Chi-Square χ². For immunolabeling it was used anti-CD4, anti-CD8, anti-FoxP3 and HSP60 monoclonal antibodies and were attributed scores from 0 to 3. Clinical-pathological relationship was analyzed using Spearman correlation. This study was approved by the Committee for Ethics in Research using Animals (CEUA-UECE), protocol 12247080-2. Our data showed a mean age of 9.3 years-old, the size of tumors presented more than 5 cm (50%), which were located in inguinal mammary glands (70%), and CMTs shows I (70%) and II (30%) grade. The cellular infiltrate was distributed both in peri and intratumoral regions, dispersed multifocally with moderate intensity and lymphocytes were the major populations found into tumors (n = 826 ± 220). In relationship to cellular infiltrate with CMT grade it was observed that lymphocytes (ρ = 0.28) and plasma cells (ρ = 0.22) showed a slight positive correlation, and an opposed negative correlation of neutrophils (ρ = -0.1) and macrophages (ρ = -0.38). CMT presents moderate lymphocytic infiltrate (< 800 lymphocytes), shows higher (P = 0.01) survival rates as compared to intense lymphocytic infiltrate (≥ 800 lymphocytes). FoxP3+ showed lower intensity while CD4+ and CD8+ expression were concentrated surrounding of lymphocytic infiltrate tumor region. HSP60+ was observed in the inflammatory and tumor cells.Discussion: Our data are according to a greater risk to the development of breast tumor in old bitches, not castrated and before or after puberty, as well as the use of contraceptives based on progesterone and estrogen. In relation to size of tumor, these findings reinforce that there is a relationship of tumor size with a higher malignancy grade and with a worse prognosis. The predominant tumor location was in the inguinal breasts that is attributed to the high activity of the mammary glands to hormonal stimuli. CMT with low clinical staging are associated with greater overall survival of affected bitches. In relation to tumor microenvironment, it has been reported that heterogeneous populations of the immune system cells often infiltrate the mammary tumors, whose lymphocytes are the main cells. It is suggested that tumor lymphocytosis may be necessary for malignant behavior of the tumor microenvironment. On the other hand, macrophages and neutrophils play an important role that may favor or inhibit the tumor cells development in the tumor microenvironment. In our work, CD4, CD8 and FoxP3 labeling were distributed in peri and intratumoral regions, and consequently, these markers can be used as prognostic for CMT, as well as being a potential target for anticancer therapies. This is the first work that presents results about the participation of HSP60 in CMT, however this data needs further investigation. HSP60 participates as a potent activator of the immune system through its peptides and other HSP types were studied in mammary carcinomas in bitches and presenting results that indicate the association of these proteins with the carcinogenesis process

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 biochemical parameters were analyzed by Kruskal-Wallis test followed by the Dunn’s test and expressed as mean ± standard deviation, with values P < 0.05. Leukocytosis (not significant), red blood cells, hematocrit and hemoglobin (P < 0.05), total protein serum (P < 0.05), globulins (P < 0.05), albumin and A/G ratio (P < 0.01) were altered in SD in relation to CD. Cutaneous cellular infiltration, composed by macrophages, plasma cells, lymphocytes and neutrophils, was observed in CD. There was an increase of expression of the markers in SD when compared to the other groups, as moderate CD68+ expression and L. infantum, and intense CD45+ and E-cadherin+ expressions.Discussion: Cutaneous involvement is very important in CanL, as it corresponds to where is the first interaction between the parasite and the immune system. Dermatological clinical signs, leukocytosis, anemia, globulins levels have been reported for dogs naturally infected by L. infantum. Inflammatory infiltrate was distributed at superficial and deep dermis, which was composed by mononuclear cells as macrophages, plasma cells, lymphocytes and neutrophils. To characterize the immune sentinels cells in the skin it was evaluated CD45+, CD68+ and E-cadherin+ expressions. In syntomatic dogs, our results revelead an increase of expression of these markers. CD45+ is one of the most abundant molecules expressed on the white blood cell surface in various mammals, while CD68+ is a myelomonocytic marker that seems to be retained during monocyte differentiation. In the skin, increased numbers of CD68+ are related to dendritic epidermal cells, which can be expressed as CD45+/CD1a-/HLA-DR+. DCs of the skin, particularly epidermal Langerhans cells (LCs), form networks anchored to neighboring keratinocytes via E-cadherin. Thus, CD45+, CD68+ and E-cadherin+ expressions may be related to activation of skin sentinels cells in dogs naturally infected by L. infantum. Our results indicated that CanL modify the CD45+, CD68+ and E-cadherin+ expressions, which characterize the immune sentinels cells activation that promove the recruitment the cellular infiltrate, which was composed by macrophages, plasma cells, lymphocytes and neutrophils. Thus, these informations may contribute to the follow-up of CanL progression in skin