Leukocytes and albumin in canine leishmaniasis

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). 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). 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 Leishmania infantum. 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.(AU)

Impact of massage on goats on the human-animal relationship and parameters linked to physiological response / Impacto da massagem em cabras na relação homem-animal e parâmetros relacionados à resposta fisiológica

ABSTRACT: Considering the benefits that massage can promote, this research aimed to apply the stroking massage in five adult goats (G1-G5) assessed in group and individually for ten days. Parameters related to massage, immune system, physiology and behavior of goats were observed. There was only significant difference (P<0.05) regarding behavior expressed in the presence of the masseur in G4 and G5, between week 1 and 2. Most goats accepted massage, except G2, with greater demonstrations of positive than negative behavior towards the masseur. Therefore, stroking massage promoted positive interactions between the masseur and goats, and should be explored as an alternative method to those existing in improving the relationship between humans and farm animals.

RESUMO: Considerando os benefícios que a massagem pode promover, este trabalho teve como objetivo aplicar a massagem do tipo stroking em cinco cabras adultas (G1-G5) avaliadas em grupo e individualmente por dez dias. Parâmetros relacionados à massagem, sistema imunológico, fisiologia e comportamento das cabras foram observados. Houve apenas diferença significativa (P<0,05) em relação ao comportamento expresso na presença da massagista em G4 e G5, entre as semanas 1 e 2. A maioria das cabras aceitou a massagem, exceto G2, com maiores demonstrações de comportamentos positivos do que negativos direcionados à massagista. Portanto, a massagem do tipo stroking promoveu interações positivas entre a massagista e as cabras, devendo ser explorada como um método alternativo aos existentes na melhora da relação entre seres humanos e animais de produção.

Adesão leucocitária e MDA sérico em cães naturalmente infectados por Leishmania infantum / Leukocyte adhesion and MDA serum levels in naturally infected dogs by Leishmania infantum

O presente estudo avaliou o estresse oxidativo e a adesão de leucócitos (AL) em cães naturalmente infectados por Leishmania infantum. Foram utilizados cães saudáveis (CN = 10) e cães acometidos por leishmaniose visceral na forma sintomática (CS = 10), submetidos previamente a exames de imunofluorescência indireta (IFI), ensaio imunoenzimático (ELISA) e pesquisa do parasito em aspirados de medula óssea. Soro foi utilizado para avaliação de malondialdeído (MDA) no ensaio para espécies reativas ao ácido tiobarbitúrico (TBARS) e AL foi determinada pelo método da coluna de náilon em sangue em EDTA, heparina e citrato. Os dados de AL foram expressos em porcentagem e MDA em média ± desvio padrão, submetidos ao teste T de student não pareado (p < 0,05). Amostras em heparina apresentaram níveis mais elevados de AL no grupo CS (55,62%, p < 0,05) quando comparadas com EDTA e citrato (10,46% e 5,28%). Citrato e EDTA inibiram AL em cães doentes e saudáveis, enquanto a heparina preservou a AL. A proporção neutrofílica se apresentou reduzida nas amostras em heparina (85% para 67%, p <0,05) quando comparadas com citrato e EDTA, que por sua vez mantiveram-se estáveis (83% para 80%). Os níveis de MDA apresentaram-se mais elevados em CS (0,0117µM ± 0,002) quando comparado com CN (0,0057µM ± 0,001) (p <0,05). Estes dados dão suporte à conclusão de que na LVC ocorre elevação do estresse oxidativo e aumento da expressão das moléculas de adesão nos leucócitos, evidenciando resposta inflamatória sistêmica. A escolha do anticoagulante é importante para a implementação do ensaio de AL.

The present study evaluated the oxidative stress and leukocyte adhesion (LA) in dogs naturally infected by Leishmania infantum. Healthy dogs (HD = 10) and dogs affected by canine visceral leishmaniasis (CVL) in symptomatic form (SD= 10) were previously submitted to indirect immunofluorescence reaction (IIF), enzyme-linked immunosorbent assay (ELISA) and detection of parasites in bone marrow aspirates. Serum was used to assess malondialdeid (MDA) by thiobarbituric acid reactive species (TBARS) assay and AL was determined by nylon column method using whole blood stored in EDTA, citrate and heparin. AL data was expressed as percentage and MDA data as mean ± standard deviation, both submitted to the unpaired student's T test (p < 0.05). Heparin samples showed higher levels of AL in CS group (55.62 %, p < 0.05) when compared with EDTA and citrate (10.46% and 5.28%). Citrate and EDTA inhibit AL in healthy and sick dogs, while heparin preserved AL in both groups. Neutrophil proportion in heparin samples were lower (85% to 67%, p <0.05) compared with citrate and EDTA, which in turn remained practically unchanged (83% to 80%). MDA levels were higher in SD (0.0117µM ± 0.002) when compared to HD (0.0057µM ± 0.001) (p <0.05). These data support the conclusion that CVL induces oxidative stress enhance and leukocyte adhesion increase, indicating systemic inflammatory response. The choice of anticoagulant is an important decision for implementing LA assays.

Clinical and laboratory alterations in dogs naturally infected by Leishmania chagasi / Alterações clínicas e laboratoriais em cães naturalmente infectados por Leishmania chagasi

INTRODUCTION: Canine visceral leishmaniasis (CVL) is a zoonotic disease with different clinical manifestations. Parasitism often occurs in bone marrow, but changes have been observed in peripheral blood and serum biochemical parameters. The aim of this study was to evaluate the hematological and biochemical parameters in dogs naturally infected by Leishmania chagasi. METHODS: Eighty-five adult dogs of both sexes and various weights and ages from the Zoonosis Control Center of Fortaleza (CCZ) were used, selected by immunofluorescence assay (IFA) and considered positive with IFA titers greater than 1:40 and by visualizing amastigotes of Leishmania chagasi in smears obtained by bone marrow aspiration. The dogs (n = 85) were grouped according to clinical signs: negative (CN = 7), subclinical (CS = 10), and clinical (CC = 68). Blood samples were collected for determination of hematological and biochemical serum values. The experimental protocol was approved by the CEUA/UECE. RESULTS: The most frequent clinical signs were cachexia (77.9 percent), keratitis (61.8 percent), and lymphadenopathy (55.9 percent), and 86.8 percent of the animals showed more than one clinical sign characteristic of CVL. In CC were observed reductions in red blood cells (63 percent), hematocrit (72 percent), and hemoglobin (62 percent), as well as leukocytosis (33 percent), neutropenia (28 percent), thrombocytopenia (50 percent), uremia (45 percent), hyperproteinemia (53 percent, p<0.05), hypergammaglobulinemia (62 percent, p<0.01), and hypoalbuminemia (58 percent). CONCLUSIONS: Animals with the clinical form of the disease demonstrate hematological and biochemical changes consistent with anemia, uremia, hyperproteinemia, and hyperglobulinemia, which present themselves as strong clinical markers of visceral leishmaniasis associated with the signs previously reported.

INTRODUÇÃO: A leishmaniose visceral canina (LVC) é uma zoonose com diferentes manifestações clínicas. O parasitismo ocorre frequentemente na medula óssea e têm sido relatadas alterações hematológicas e bioquímicas. Objetivou-se avaliar os parâmetros clínicos, hematológicos e bioquímicos de cães naturalmente infectados por Leishmania chagasi. MÉTODOS: Utilizaram-se 85 cães adultos, ambos os sexos, peso e idade variados, oriundos do Centro de Controle de Zoonoses de Fortaleza, selecionados pela reação de imunofluorescência indireta (RIFI), sendo considerados positivos os animais com títulos de RIFI > 1:40 e pelo exame parasitológico das formas amastigotas de Leishmania chagasi em esfregaços de medula óssea. Os cães foram agrupados conforme os sinais clínicos associados à doença: negativos (CN=7); subclínicos (CS=10) e clínicos (CC=68). Amostras de sangue foram coletadas para determinação dos parâmetros hematológicos e bioquímicos séricos. O protocolo experimental foi aprovado pelo CEUA/UECE, protocolo n° 08622833-1. RESULTADOS: Os sinais clínicos mais frequentes foram caquexia (77,9 por cento), ceratoconjuntivite (61,8 por cento) e linfadenopatia (55,9 por cento), sendo que 86, 8 por cento dos animais apresentaram mais de um sinal clínico característico de LVC. Em CC foram observadas reduções nas hemácias (63 por cento), hematócrito (72 por cento) e hemoglobina (62 por cento), leucocitose (33 por cento), neutropenia (28 por cento), trombocitopenia (50 por cento), uremia (45 por cento), hiperproteinemia (53 por cento, p<0,05), hiperglobulinemia (62 por cento, p<0,01) e hipoalbuminemia (58 por cento). CONCLUSÕES: Concluiu-se que os animais com a forma clínica da doença apresentam alterações condizentes com anemia, uremia, hiperproteinemia e hiperglobulinemia, as quais se apresentam como marcadores clínicos da leishmaniose visceral, associados aos sinais previamente relatados.