High seroprevalence and associated factors for visceral leishmaniasis in dogs in a transmission area of Paraíba state, Northeastern Brazil.

Dogs have been implicated as main reservoirs for visceral leishmaniasis (VL) in urban areas. Therefore, studies on this species provide important data for public health. Thus, the objective of the present study was to ascertain the seroprevalence of canine VL (CVL) and the associated factors in the Brejo Paraibano microregion, northeastern Brazil. A total of 409 dogs were sampled from the eight municipalities of the microregion: Alagoa Grande, Alagoa Nova, Areia, Bananeiras, Borborema, Matinhas, Pilões and Serraria. The diagnosis of CVL was made using an enzyme-linked immunosorbent assay (ELISA S7®), through which the prevalence observed was 29.3%. By robust Poisson regression analysis two factors were associated with seroprevalence: illiterate/incomplete elementary owner's education level (prevalence ratio = 1.57; 95% CI = 1.05-2.34; p-value = 0.027) and tick infestation (prevalence ratio = 1.82; 95% CI = 1.27-2.61; p-value = 0.001). It is concluded that the seroprevalence of CVL in the Brejo Paraibano microregion is high. The factors associated with seroprevalence indicated the importance to develop socioeducational actions on the population, and the finding that tick infestation was associated with seroprevalence shows that there is a need for investigation regarding the role of ticks in the epidemiology of CVL.

Desempenho de diferentes marcas de tiras reagentes relativo à densidade e à análise química da urina de cães e gatos / Performance of different brands of reagent strips for density and chemical analysis of dog and cat urine

Objetivou-se verificar a compatibilidade entre diferentes marcas de tiras reagentes para urinálise, tanto de uso veterinário, como de uso humano, e confrontar os parâmetros semiquantitativos desse instrumento com métodos quantitativos. Para isso, foram analisadas 77 amostras frescas de urina de cães e gatos e testados 04 modelos de tiras reagentes. Quanto à densidade urinária, houve correlação razoável entre os métodos quantitativo e semiquantitativo naquelas amostras com pH ácido, mas não naquelas com pH neutro ou alcalino. Quanto à concentração proteica, houve similaridade de 53,3% a 83,3% entre as marcas testadas e quando comparadas com a análise fotométrica houve uma correlação razoável (rs = 0,69752 a 0,75074). Em ponto de corte de 15mg/dL de proteína, a sensibilidade da tira reagente foi 82,5% e 100% para urina canina e felina, respectivamente. No tocante à hematúria, houve divergência razoável entre a sedimentoscopia e as diferentes marcas de tiras reativas. Quanto à piúria, há uma baixa sensibilidade das tiras em relação às amostras caninas com muitos resultados falso-negativos (33% a 75%), enquanto em amostras felinas a sensibilidade foi de 100%. Assim, independente da marca, as tiras reagentes devem servir apenas como teste rápido de triagem, sendo mais apropriado o uso de métodos quantitativos na avaliação clínica do paciente a partir da urinálise.

The aim was to verify the compatibility between different brands of urinary dipsticks, for both human and veterinary use, and to compare the semiquantitative parameters of this instrument with quantitative methods. For this, 77 fresh samples of urine from dogs and cats were analyzed e and 04 models of reagent strips were tested. Regarding urinary density, a reasonable correlation was observed between the quantitative and semiquantitative methods in those samples with acidic pH, which did not occur in those with neutral or alkaline pH. Regarding the protein concentration, there was similarity from 53.3% to 83.3% between the brands and in the comparative analysis between the control strip and the photometric analysis, there was a reasonable correlation (rs = 0.69752 to 0.75074). In cut-off point of 15mg/dL protein, the sensitivity of the reagent strip was 82.5% and 100% for canine and feline urine, respectively. Regarding hematuria, there was a reasonable divergence of results between sedimentation and tested dipsticks. As for pyuria, there is a low sensitivity of the strips in relation to canine samples with many false negative results (33% to 75%), while in feline samples the sensitivity was 100%. Thus, regardless of the brands, the reagent strips should serve only as a rapid screening test, while the use of quantitative methods in the clinical evaluation of the patient from urinalysis is more appropriate.

Desempenho de diferentes marcas de tiras reagentes relativo à densidade e à análise química da urina de cães e gatos / Performance of different brands of reagent strips for density and chemical analysis of dog and cat urine

Objetivou-se verificar a compatibilidade entre diferentes marcas de tiras reagentes para urinálise, tanto de uso veterinário, como de uso humano, e confrontar os parâmetros semiquantitativos desse instrumento com métodos quantitativos. Para isso, foram analisadas 77 amostras frescas de urina de cães e gatos e testados 04 modelos de tiras reagentes. Quanto à densidade urinária, houve correlação razoável entre os métodos quantitativo e semiquantitativo naquelas amostras com pH ácido, mas não naquelas com pH neutro ou alcalino. Quanto à concentração proteica, houve similaridade de 53,3% a 83,3% entre as marcas testadas e quando comparadas com a análise fotométrica houve uma correlação razoável (rs = 0,69752 a 0,75074). Em ponto de corte de 15mg/dL de proteína, a sensibilidade da tira reagente foi 82,5% e 100% para urina canina e felina, respectivamente. No tocante à hematúria, houve divergência razoável entre a sedimentoscopia e as diferentes marcas de tiras reativas. Quanto à piúria, há uma baixa sensibilidade das tiras em relação às amostras caninas com muitos resultados falso-negativos (33% a 75%), enquanto em amostras felinas a sensibilidade foi de 100%. Assim, independente da marca, as tiras reagentes devem servir apenas como teste rápido de triagem, sendo mais apropriado o uso de métodos quantitativos na avaliação clínica do paciente a partir da urinálise.

The aim was to verify the compatibility between different brands of urinary dipsticks, for both human and veterinary use, and to compare the semiquantitative parameters of this instrument with quantitative methods. For this, 77 fresh samples of urine from dogs and cats were analyzed e and 04 models of reagent strips were tested. Regarding urinary density, a reasonable correlation was observed between the quantitative and semiquantitative methods in those samples with acidic pH, which did not occur in those with neutral or alkaline pH. Regarding the protein concentration, there was similarity from 53.3% to 83.3% between the brands and in the comparative analysis between the control strip and the photometric analysis, there was a reasonable correlation (rs = 0.69752 to 0.75074). In cut-off point of 15mg/dL protein, the sensitivity of the reagent strip was 82.5% and 100% for canine and feline urine, respectively. Regarding hematuria, there was a reasonable divergence of results between sedimentation and tested dipsticks. As for pyuria, there is a low sensitivity of the strips in relation to canine samples with many false negative results (33% to 75%), while in feline samples the sensitivity was 100%. Thus, regardless of the brands, the reagent strips should serve only as a rapid screening test, while the use of quantitative methods in the clinical evaluation of the patient from urinalysis is more appropriate.

Prednisolone associated with doxycycline on the hematological parameters and serum proteinogram of dogs with ehrlichiosis / Efeitos da influência da prednisolona associada à doxiciclina nos parâmetros hematológicos e proteinograma sérico de cães com erliquiose

ABSTRACT: The objective of the present study was to assess the treatment of dogs with ehrlichiosis (tropical canine pancytopenia (TCP)) with doxycycline hydrochloride associated or not with prednisolone on the hematological profile and serum proteins. Ten dogs with TCP were selected in the Small Animal Medical Clinic Veterinary Hospital, Federal University of Campina Grande-UFCG-PB, Brazil. The diagnosis was obtained by clinical examination, hemogram and immunochromatographic test (with anti Ehrlichia canis antibodies). Samples were distributed randomly in two experimental groups of five animals each (n = 5), named GD and GDP. The GD group was treated with doxycycline (5 mg/kg, VO, BID for 28 days) and the GDP group was treated with doxycycline at the same dose and duration and prednisolone (2 mg/kg, VO, BID, for five days). Four blood collections were made during the treatment period: a base collection (M0), one at 10 days (M10), a second at 21 days (M21) and another at the end of the treatment (M28). These samples were used for the tests erythrogram, leucogram, plateletgram and proteinogram (dose of total proteins, pre-albumin, albumin, albumin, α-1 globulin, α-2 globulin, ß-globulin, ɣ-globulin and C- reactive protein) in the Clinical Pathology Laboratory, Veterinary Hospital-UFCG-PB. Results, that presented normal distribution, was submitted to the Tukey test (P<0.05). Comparison of treatments GD and GDP showed that both promoted discreet and similar response in the hematological parameters at different times. Results obtained allowed the conclusion that both therapeutic protocols resulted in clinical, hematological parameter and proteinogram improvement, but the use of prednisolone at the dose administered during the first five days of treatment did not show more beneficial effects than isolated administration of doxycycline.

RESUMO: Objetivou-se com este estudo avaliar os efeitos do tratamento de cães com erliquiose monocítica canina (EMC) com cloridrato de doxiciclina associada ou não à prednisolona sobre o perfil hematológico e de proteínas séricas. Foram selecionados, na Clínica Médica de Pequenos Animais do Hospital Veterinário da Universidade Federal de Campina Grande (UFCG), 10 cães com EMC. O diagnóstico foi obtido através de exame clínico, hemograma e teste de imunoensaio imunocromatográfico (com anticorpos anti Ehrlichia canis). As amostras foram distribuídas aleatoriamente em dois grupos experimentais, com cinco animais cada (n=5), denominados GD e GDP. O grupo GD foi tratado com doxiciclina (5 mg/kg, VO, BID durante 28 dias) e o grupo GDP, tratado com doxiciclina na mesma dose e duração e prednisolona (2 mg/kg, VO, BID, durante 5 dias). Durante o período de tratamento, foram realizadas quatro coletas de sangue; uma coleta basal (M0), uma com 10 dias (M10), uma aos 21 dias (M21) e outra ao final do tratamento (M28). Dessas amostras foram realizados eritrograma, leucograma, plaquetograma e proteinograma (dosagem de proteínas totais, pré-albumina, albumina, α-1 globulina, α-2 globulina, ß-globulina, ɣ-globulina e proteína C-reativa) no Laboratório de Patologia Clínica do Hospital Veterinário da UFCG. Os resultados que apresentaram distribuição normal foram submetidos ao teste de Tukey (P<0,05). Ao comparar os tratamentos GD e GDP, observou-se que os dois promoveram resposta discreta e semelhante dos parâmetros hematológicos nos diferentes momentos. Os resultados obtidos permitem concluir que ambos os protocolos terapêuticos resultaram em melhora clínica e dos parâmetros hematológicos e do proteinograma. Porém, o uso da prednisolona na dose empregada durante os primeiros cinco dias de tratamento não demonstrou efeitos mais benéficos do que a administração isolada da doxiciclina.

Purification, characterization and antibacterial potential of a lectin isolated from Apuleia leiocarpa seeds.

Apuleia leiocarpa is a tree found in Caatinga that has great value in the timber industry. Lectins are carbohydrate-binding proteins with several biotechnological applications. This study shows the isolation, characterization, and antibacterial activity of A. leiocarpa seed lectin (ApulSL). The lectin was chromatographically isolated from a crude extract (in 150 mM NaCl) by using a chitin column. ApulSL adsorbed to the matrix and was eluted using 1.0 M acetic acid. Native ApulSL was characterized as a 55.8-kDa acidic protein. SDS-PAGE showed three polypeptide bands, whereas two-dimensional electrophoresis revealed four spots. The peptides detected by MALDI TOF/TOF did not show sufficient homology (<30%) with the database proteins. Circular dichroism spectroscopy suggested a disordered conformational structure, and fluorescence spectrum showed the presence of tyrosine residues in the hydrophobic core. The hemagglutinating activity of ApulSL was present even after heating to 100 °C, was Mn(2+)-dependent, and inhibited by N-acetylglucosamine, D(-)-arabinose, and azocasein. ApulSL demonstrated bacteriostatic and bactericide effects on gram-positive and gram-negative species, being more effective against three varieties of Xanthomonas campestris (MIC ranging from 11.2 to 22.5 µg/mL and MBC of 22.5 µg/mL). The results of this study reinforce the importance of biochemical prospecting of Caatinga by revealing the antibacterial potential of ApulSL.

Purification of a lectin with antibacterial activity from Bothrops leucurus snake venom.

A novel lectin was isolated from Bothrops leucurus snake venom using a combination of affinity and gel filtration chromatographies. The lectin (BlL) agglutinated glutaraldehyde-treated rabbit and human erythrocytes with preference for rabbit erythrocytes. Galactose, raffinose, lactose, fetal bovine serum and casein inhibited lectin-induced rabbit erythrocyte agglutination. BlL, with a molecular mass of 30 kDa and composed of two subunits of 15 kDa, showed dependence on calcium. BlL is an acidic protein with highest activity over the pH range of 4.0-7.0 and stable under heating to 70°C. Fluorescence emission spectra showed tryptophan residues partially buried within the lectin structure. The percentages of secondary structure revealed by circular dichroism were 1% α-helix, 44% ß-sheet, 24% ß-turn and 31% unordered. BlL showed effective antibacterial activity against Gram-positive bacteria Staphylococcus aureus, Enterococcus faecalis and Bacillus subtilis with minimal inhibitory concentrations of 31.25, 62.25 and 125 µg/mL, respectively. In conclusion, B. leucurus snake venom contains a galactoside-binding lectin with antibacterial activity.