Daniela Larangeira - Academia.edu (original) (raw)

Papers by Daniela Larangeira

Research paper thumbnail of A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: Assessment of leukocyte population changes in canine visceral leishmaniosis

A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirat... more A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB + cells were higher, and of CD45RA + cells were lower, both in the spleen and in the blood of animals with www.elsevier.com/locate/vetimm Veterinary Immunology and Immunopathology 111 (2006) 251-261 leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy. #

Research paper thumbnail of JVAC14959

Research paper thumbnail of Investigation of risk areas as complemental methodology for the control of canine visceral leishmaniasis

Pesquisa Veterinária Brasileira

Risk areas of canine visceral leishmaniasis in the city of Camaçari, Bahia, Brazil, were investig... more Risk areas of canine visceral leishmaniasis in the city of Camaçari, Bahia, Brazil, were investigated. A total of 278 dogs from 141 homes pertaining to 20 investigated risk areas was serologically screened (ELISA). The general seroprevalence was 21.7% (56/258) after exclusion of 20 dogs used at the beginning of the survey to limit the study area. The respective results of the univariated and multivariated analysis of factors related to infection of dogs by Leishmania chagasi, to vector distribu-tion pattern in the area and to the methodology used to localize the canine focuses are discussed.

Research paper thumbnail of Severe Clinical Presentation of Visceral Leishmaniasis in Naturally Infected Dogs with Disruption of the Splenic White Pulp

PLoS ONE, 2014

In this work, we investigated the association between the disruption of splenic lymphoid tissue a... more In this work, we investigated the association between the disruption of splenic lymphoid tissue and the severity of visceral leishmaniasis in dogs. Clinical and laboratory data from 206 dogs were reviewed. Spleen sections collected during the euthanasia of these animals were analyzed, and the splenic lymphoid tissue samples were

Research paper thumbnail of Characterization of clinical, immunological and parasitological parameters during a steady state of improvement of after chemotherapy of Leishmania chagasi naturally infected dogs

Veterinary Immunology and Immunopathology, 2009

the strains LLM-759 and LLM-480 did not showed clinical, immunological and parasitological status... more the strains LLM-759 and LLM-480 did not showed clinical, immunological and parasitological status of disease along the follow up. However, the dogs inoculated with the strain LLM-724 showed alterations in all the parameters studied and a parasitological positive status one year after infection. In order to test the protective capability of these new variants of L. infantum, those dogs infected with the strains LLM-759 and LLM-480 were challenged intravenously with the virulent strain and monitored during 8 months. Leishmania-specific serum antibodies and presence of the parasite was confirmed in those animals previously infected with LLM-480 6 months after challenge with the virulent strain, while dogs infected with LLM-759 remained parasitologically negative. These results confirm that new L. infantum variants LLM-759 and LLM-480 are not able to induce the outcome of the disease in the canine host and the possible protective capability of the strain LLM-759.

Research paper thumbnail of Investigação de áreas de risco como metodologia Investigação de áreas de risco como metodologia Investigação de áreas de risco como metodologia complementar ao controle da leishmaniose visceral canina complementar ao controle da leishmaniose visceral canina complementar ao controle da leishmanios...

Research paper thumbnail of A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: Assessment of leukocyte population changes in canine visceral leishmaniosis

Veterinary Immunology and Immunopathology, 2006

A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirat... more A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB + cells were higher, and of CD45RA + cells were lower, both in the spleen and in the blood of animals with www.elsevier.com/locate/vetimm Veterinary Immunology and Immunopathology 111 (2006) 251-261 leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy. #

Research paper thumbnail of Comparison of two commercial vaccines against visceral leishmaniasis in dogs from endemic areas: IgG, and subclasses, parasitism, and parasite transmission by xenodiagnosis

Vaccine, 2014

Background: The incidence of zoonotic canine visceral leishmaniasis (CVL) would decrease if dogs ... more Background: The incidence of zoonotic canine visceral leishmaniasis (CVL) would decrease if dogs were effectively vaccinated; however, additional data on the efficacy of canine vaccines are required for their approved preventative use.

Research paper thumbnail of Can spleen aspirations be safely used for the parasitological diagnosis of canine visceral leishmaniosis? A study on assymptomatic and polysymptomatic animals

The Veterinary Journal, 2006

The purpose of this study was to evaluate the safety of spleen aspiration as a sampling technique... more The purpose of this study was to evaluate the safety of spleen aspiration as a sampling technique for the parasitological detection by culture and microscopy of Leishmania (chagasi) infantum. Two hundred and nine domiciled dogs from an endemic area for visceral leishmaniasis in Bahia State, Brazil, were studied. Most dogs (87%) were seropositive for anti-L. chagasi antibodies by ELISA. Clinical signs of disease were recorded and the animals monitored during and after spleen puncture in order to detect possible complications associated with the procedure. From a total of 257 splenic punctures in the 209 animals, only three minor events occurred, with no significant consequence for the animals and no association with risk factors. Leishmania was isolated from 149/180 (83%) seropositive dogs, and from 6/26 (23%) seronegative animals. The procedure did not cause adverse side effects or unnecessary suffering and confirmed the diagnosis in a large percentage of dogs. We conclude that spleen aspiration can be considered an effective and safe procedure for the definitive diagnosis of canine visceral leishmaniosis.

Research paper thumbnail of Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres

Pesquisa Veterinária Brasileira, 2013

Pesq. Vet. Bras. 33(4):528-534, abril 2013 528 RESUMO.-Na América do Sul, alguns canídeos silvest... more Pesq. Vet. Bras. 33(4):528-534, abril 2013 528 RESUMO.-Na América do Sul, alguns canídeos silvestres são considerados reservatórios naturais de Leishmania chagasi. A resposta imunológica desses animais a Leishma-Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres 1 -Melo S.M. 2013. [Indirect ELISA for the serological diagnosis of visceral leishmaniasis in wild canids.] Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres. Pesquisa Veterinária Brasileira 33(4):528-534.

Research paper thumbnail of Investigação de áreas de risco como metodologia complementar ao controle da leishmaniose visceral canina

Pesquisa Veterinária Brasileira, 2007

Research paper thumbnail of Eletrocardiografia computadorizada em cães: estudo comparativo

Pesquisa Veterinária Brasileira, 2013

Pesq. Vet. Bras. 33 :949-953, julho 2013 949 RESUMO.-O método de eletrocardiografia computadoriza... more Pesq. Vet. Bras. 33 :949-953, julho 2013 949 RESUMO.-O método de eletrocardiografia computadorizada (ECG-C) vem sendo crescentemente difundido na medicina veterinária, havendo atualmente diversas marcas e modelos de eletrocardiógrafos disponíveis no mercado. Diante da possibilidade de diferenças na sensibilidade e na reprodutibilidade das medidas obtidas nos traçados, o presente estudo teve como objetivo comparar os parâmetros eletrocardiográficos de cães, obtidos por dois siste-ABSTRACT.-Oliveira L.S., Santos R.R.B., Melo M.B., Laranjeira D.F. & Barrouin-Melo S.M. 2013. [Computerized electrocardiography in dogs: a comparative study.] Eletrocardiografia computadorizada em cães: estudo comparativo. Pesquisa Veterinária Brasileira 33 :949-953. Computerized electrocardiography (C-EKG) has been more frequently used in Veterinary Medicine. Many equipment models are available for this purpose. Due to possible device sensitivity and reproducibility differences during examination, the main goal of this study was to compare electrocardiographic parameters of dogs using two different C-EKG systems: Wincardio Micromed ® (WIN) and TEB ECGPC ® (TEB). Forty two healthy male and female dogs of different breeds (Cocker Spaniel, Dachshund, Labrador, Pinscher, Pitbull Terrier, Poodle, Schnauzer, Shih Tzu, Yorkshire and mongrel dogs), with age between 4 months and 16 years old were grouped according to weight and evaluated by both systems. The electrocardiographic measurements were performed on DII lead for both systems. The study showed that the TEB system was more sensitive for measurement of P wave and QRS complex duration, while the WIN system showed more sensitivity for the measurements of amplitude of the same parameters. The larger animals (26-37kg) showed greater variance in the measurements of P wave and QRS complex amplitude and duration than the groups of medium (14-25kg) or smaller (3-13kg) dogs. These differences must be considered when using diverse computerized electrocardiography systems to perform measurements due to the possibility of erratic interpretation of the results between veterinary medicine services.

Research paper thumbnail of Comparison between splenic and lymph node aspirations as sampling methods for the parasitological detection of Leishmania chagasi infection in dogs

Memórias do Instituto Oswaldo Cruz, 2004

The sensitivities of spleen and lymph node cultures for the diagnosis of canine visceral leishman... more The sensitivities of spleen and lymph node cultures for the diagnosis of canine visceral leishmaniasis were compared in 64 anti-Leishmania antibody positive dogs from an endemic area in Brazil. The sensitivity of spleen cultures for Leishmania detection was 97.9%; in lymph node cultures it was 25%. Positive spleen culture was more frequent (p = 0.048, Fisher's exact probability test) in symptomatic (28 out of 33 animals) than in asymptomatic animals (19 out of 31 animals). These results support the use of spleen instead of lymph node aspiration as the choice method for the parasitological diagnosis of the infection.

Research paper thumbnail of Enhancement of Experimental Cutaneous Leishmaniasis by Leishmania Molecules Is Dependent on Interleukin-4, Serine Protease/Esterase Activity, and Parasite and Host Genetic Backgrounds

Infection and Immunity, 2011

Most inbred strains of mice, like the BALB/c strain, are susceptible to Leishmania amazonensis in... more Most inbred strains of mice, like the BALB/c strain, are susceptible to Leishmania amazonensis infections and resistant to Leishmania braziliensis infections. This parasite-related difference could result from the activity of an L. amazonensis-specific virulence factor. In agreement with this hypothesis, it is shown here that the intravenous injection of BALB/c mice with L. amazonensis amastigote extract (LaE) but not the L. braziliensis extract confers susceptibility to L. braziliensis infection. This effect was associated with high circulating levels of IgG1 anti-L. amazonensis antibodies and with an increase in interleukin-4 (IL-4) production and a decrease in gamma interferon production by draining lymph node cells. Moreover, the effect was absent in IL-4knockout mice. The biological activity in the LaE was not mediated by amphiphilic molecules and was inhibited by pretreatment of the extract with irreversible serine protease inhibitors. These findings indicate that the LaE contains a virulence-related factor that (i) enhances the Leishmania infection by promoting Th2-type immune responses, (ii) is not one of the immunomodulatory Leishmania molecules described so far, and (iii) is either a serine protease or has an effect that depends on that protease activity. In addition to being Leishmania species specific, the infection-enhancing activity was also shown to depend on the host genetic makeup, as LaE injections did not affect the susceptibility of C57BL/6 mice to L. braziliensis infection. The identification of Leishmania molecules with infection-enhancing activity could be important for the development of a vaccine, since the up-or downmodulation of the immune response against a virulence factor could well contribute to controlling the infection.

Research paper thumbnail of Production of Monoclonal Antibodies Against Canine Leukocytes

Hybridoma and Hybridomics, 2004

A panel of anti-canine leukocyte monoclonal antibodies (MAbs) was produced by immunizing BALB/c m... more A panel of anti-canine leukocyte monoclonal antibodies (MAbs) was produced by immunizing BALB/c mice with canine peripheral blood mononuclear cells (PBMC), either resting or stimulated with concanavalin A (ConA). Three out of 28 clones—IH1, AB6, and HG6–screened by ...

Research paper thumbnail of A Novel Monoclonal Antibody Against Canine Monocytes/Macrophages

Hybridoma and Hybridomics, 2004

The production and partial characterization of a monoclonal antibody, the IgG 1 IH1, which recogn... more The production and partial characterization of a monoclonal antibody, the IgG 1 IH1, which recognizes an antigen distributed in canine monocytes/macrophages, is reported here. The distribution and apparent molecular weight of the antigen recognized by the IH1 MAb was determined in peripheral blood leukocytes, peripheral blood monocyte-derived macrophages and tissue sections of spleen, liver and skin, using Western blotting, immunocytochemistry, immunohistochemistry and flow cytometry. The IH1 MAb-recognized antigen was detected in Western blotting under non-reducing conditions spread out as a large band covering the position corresponding to the migration of molecules with molecular weights from 55 to 73 kDa. The IH1 MAb labeled blood monocytes, tissue macrophages in lymph nodes, and in the mantle zone of the spleen, and Kupffer cells in the liver. It did not react with human cells. In flow cytometric analysis, the IH1 MAb reacted with a subpopulation of monocytes. The MAb described herein may become a valuable tool for diagnosis and research on canine diseases.

Research paper thumbnail of Ectopia ureteral unilateral congênita em uma cadela Teckel Dachshund com pelagem arlequim: relato de caso

Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 2012

O presente relato apresenta a primeira descrição de ectopia ureteral congênita na raça Teckel Dac... more O presente relato apresenta a primeira descrição de ectopia ureteral congênita na raça Teckel Dachshund, diagnosticada em uma cadela com pelagem arlequim. O animal, aos dois meses de idade, apresentava sinais de incontinência urinária e cistite bacteriana, sendo submetido a um plano diagnóstico para confirmação de ureter ectópico. A urografia excretora revelou hidroureter direito com desembocadura caudal ao trígono da bexiga. O exame físico do animal evidenciou ainda hérnia inguinal bilateral, o que reforçou a caracterização da origem congênita das alterações. Aspectos de bem-estar animal são também discutidos.

Research paper thumbnail of An Assessment of the Genetic Diversity of Leishmania infantum Isolates from Infected Dogs in Brazil

American Journal of Tropical Medicine and Hygiene, 2012

Correlations between the genetic diversity of Leishmania infantum (syn. L. chagasi)isolatesandthe... more Correlations between the genetic diversity of Leishmania infantum (syn. L. chagasi)isolatesandtheirrespective geographic origins support the theoretic assumption that visceral leishmaniasis probably originated in the Old World. Because dogs are widely considered to be the main reservoir of this disease, the present study aimed to investigate the degree of genetic divergence among 44 leishmanial canine isolates from two Brazilian cities, Jequié and Campo Grande, located approximately 2,028 km from each other. We hypothesized that a low degree of genetic divergence would be observed among these isolates. In fact, statistical analyses found no significant differences between the isolates using both random amplified polymorphic DNA and multilocus microsatellite typing genotyping techniques with three and seven markers, respectively. These findings provide support for the recent introduction of L. infantum into the New World.

Research paper thumbnail of Evaluating the Accuracy of Molecular Diagnostic Testing for Canine Visceral Leishmaniasis Using Latent Class Analysis

PLoS ONE, 2014

Host tissues affected by Leishmania infantum have differing degrees of parasitism. Previously, th... more Host tissues affected by Leishmania infantum have differing degrees of parasitism. Previously, the use of different biological tissues to detect L. infantum DNA in dogs has provided variable results. The present study was conducted to evaluate the accuracy of molecular diagnostic testing (qPCR) in dogs from an endemic area for canine visceral leishmaniasis (CVL) by determining which tissue type provided the highest rate of parasite DNA detection. Fifty-one symptomatic dogs were tested for CVL using serological, parasitological and molecular methods. Latent class analysis (LCA) was performed for accuracy evaluation of these methods. qPCR detected parasite DNA in 100% of these animals from at least one of the following tissues: splenic and bone marrow aspirates, lymph node and skin fragments, blood and conjunctival swabs. Using latent variable as gold standard, the qPCR achieved a sensitivity of 95.8% (CI 90.4-100) in splenic aspirate; 79.2% (CI 68-90.3) in lymph nodes; 77.3% (CI 64.5-90.1) in skin; 75% (CI 63.1-86.9) in blood; 50% (CI 30-70) in bone marrow; 37.5% (CI 24.2-50.8) in left-eye; and 29.2% ) in right-eye conjunctival swabs. The accuracy of qPCR using splenic aspirates was further evaluated in a random larger sample (n = 800), collected from dogs during a prevalence study. The specificity achieved by qPCR was 76.7% (CI 73.7-79.6) for splenic aspirates obtained from the greater sample. The sensitivity accomplished by this technique was 95% (CI 93.5-96.5) that was higher than those obtained for the other diagnostic tests and was similar to that observed in the smaller sampling study. This confirms that the splenic aspirate is the most effective type of tissue for detecting L. infantum infection. Additionally, we demonstrated that LCA could be used to generate a suitable gold standard for comparative CVL testing.

Research paper thumbnail of A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: Assessment of leukocyte population changes in canine visceral leishmaniosis

A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirat... more A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB + cells were higher, and of CD45RA + cells were lower, both in the spleen and in the blood of animals with www.elsevier.com/locate/vetimm Veterinary Immunology and Immunopathology 111 (2006) 251-261 leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy. #

Research paper thumbnail of JVAC14959

Research paper thumbnail of Investigation of risk areas as complemental methodology for the control of canine visceral leishmaniasis

Pesquisa Veterinária Brasileira

Risk areas of canine visceral leishmaniasis in the city of Camaçari, Bahia, Brazil, were investig... more Risk areas of canine visceral leishmaniasis in the city of Camaçari, Bahia, Brazil, were investigated. A total of 278 dogs from 141 homes pertaining to 20 investigated risk areas was serologically screened (ELISA). The general seroprevalence was 21.7% (56/258) after exclusion of 20 dogs used at the beginning of the survey to limit the study area. The respective results of the univariated and multivariated analysis of factors related to infection of dogs by Leishmania chagasi, to vector distribu-tion pattern in the area and to the methodology used to localize the canine focuses are discussed.

Research paper thumbnail of Severe Clinical Presentation of Visceral Leishmaniasis in Naturally Infected Dogs with Disruption of the Splenic White Pulp

PLoS ONE, 2014

In this work, we investigated the association between the disruption of splenic lymphoid tissue a... more In this work, we investigated the association between the disruption of splenic lymphoid tissue and the severity of visceral leishmaniasis in dogs. Clinical and laboratory data from 206 dogs were reviewed. Spleen sections collected during the euthanasia of these animals were analyzed, and the splenic lymphoid tissue samples were

Research paper thumbnail of Characterization of clinical, immunological and parasitological parameters during a steady state of improvement of after chemotherapy of Leishmania chagasi naturally infected dogs

Veterinary Immunology and Immunopathology, 2009

the strains LLM-759 and LLM-480 did not showed clinical, immunological and parasitological status... more the strains LLM-759 and LLM-480 did not showed clinical, immunological and parasitological status of disease along the follow up. However, the dogs inoculated with the strain LLM-724 showed alterations in all the parameters studied and a parasitological positive status one year after infection. In order to test the protective capability of these new variants of L. infantum, those dogs infected with the strains LLM-759 and LLM-480 were challenged intravenously with the virulent strain and monitored during 8 months. Leishmania-specific serum antibodies and presence of the parasite was confirmed in those animals previously infected with LLM-480 6 months after challenge with the virulent strain, while dogs infected with LLM-759 remained parasitologically negative. These results confirm that new L. infantum variants LLM-759 and LLM-480 are not able to induce the outcome of the disease in the canine host and the possible protective capability of the strain LLM-759.

Research paper thumbnail of Investigação de áreas de risco como metodologia Investigação de áreas de risco como metodologia Investigação de áreas de risco como metodologia complementar ao controle da leishmaniose visceral canina complementar ao controle da leishmaniose visceral canina complementar ao controle da leishmanios...

Research paper thumbnail of A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: Assessment of leukocyte population changes in canine visceral leishmaniosis

Veterinary Immunology and Immunopathology, 2006

A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirat... more A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB + cells were higher, and of CD45RA + cells were lower, both in the spleen and in the blood of animals with www.elsevier.com/locate/vetimm Veterinary Immunology and Immunopathology 111 (2006) 251-261 leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy. #

Research paper thumbnail of Comparison of two commercial vaccines against visceral leishmaniasis in dogs from endemic areas: IgG, and subclasses, parasitism, and parasite transmission by xenodiagnosis

Vaccine, 2014

Background: The incidence of zoonotic canine visceral leishmaniasis (CVL) would decrease if dogs ... more Background: The incidence of zoonotic canine visceral leishmaniasis (CVL) would decrease if dogs were effectively vaccinated; however, additional data on the efficacy of canine vaccines are required for their approved preventative use.

Research paper thumbnail of Can spleen aspirations be safely used for the parasitological diagnosis of canine visceral leishmaniosis? A study on assymptomatic and polysymptomatic animals

The Veterinary Journal, 2006

The purpose of this study was to evaluate the safety of spleen aspiration as a sampling technique... more The purpose of this study was to evaluate the safety of spleen aspiration as a sampling technique for the parasitological detection by culture and microscopy of Leishmania (chagasi) infantum. Two hundred and nine domiciled dogs from an endemic area for visceral leishmaniasis in Bahia State, Brazil, were studied. Most dogs (87%) were seropositive for anti-L. chagasi antibodies by ELISA. Clinical signs of disease were recorded and the animals monitored during and after spleen puncture in order to detect possible complications associated with the procedure. From a total of 257 splenic punctures in the 209 animals, only three minor events occurred, with no significant consequence for the animals and no association with risk factors. Leishmania was isolated from 149/180 (83%) seropositive dogs, and from 6/26 (23%) seronegative animals. The procedure did not cause adverse side effects or unnecessary suffering and confirmed the diagnosis in a large percentage of dogs. We conclude that spleen aspiration can be considered an effective and safe procedure for the definitive diagnosis of canine visceral leishmaniosis.

Research paper thumbnail of Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres

Pesquisa Veterinária Brasileira, 2013

Pesq. Vet. Bras. 33(4):528-534, abril 2013 528 RESUMO.-Na América do Sul, alguns canídeos silvest... more Pesq. Vet. Bras. 33(4):528-534, abril 2013 528 RESUMO.-Na América do Sul, alguns canídeos silvestres são considerados reservatórios naturais de Leishmania chagasi. A resposta imunológica desses animais a Leishma-Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres 1 -Melo S.M. 2013. [Indirect ELISA for the serological diagnosis of visceral leishmaniasis in wild canids.] Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres. Pesquisa Veterinária Brasileira 33(4):528-534.

Research paper thumbnail of Investigação de áreas de risco como metodologia complementar ao controle da leishmaniose visceral canina

Pesquisa Veterinária Brasileira, 2007

Research paper thumbnail of Eletrocardiografia computadorizada em cães: estudo comparativo

Pesquisa Veterinária Brasileira, 2013

Pesq. Vet. Bras. 33 :949-953, julho 2013 949 RESUMO.-O método de eletrocardiografia computadoriza... more Pesq. Vet. Bras. 33 :949-953, julho 2013 949 RESUMO.-O método de eletrocardiografia computadorizada (ECG-C) vem sendo crescentemente difundido na medicina veterinária, havendo atualmente diversas marcas e modelos de eletrocardiógrafos disponíveis no mercado. Diante da possibilidade de diferenças na sensibilidade e na reprodutibilidade das medidas obtidas nos traçados, o presente estudo teve como objetivo comparar os parâmetros eletrocardiográficos de cães, obtidos por dois siste-ABSTRACT.-Oliveira L.S., Santos R.R.B., Melo M.B., Laranjeira D.F. & Barrouin-Melo S.M. 2013. [Computerized electrocardiography in dogs: a comparative study.] Eletrocardiografia computadorizada em cães: estudo comparativo. Pesquisa Veterinária Brasileira 33 :949-953. Computerized electrocardiography (C-EKG) has been more frequently used in Veterinary Medicine. Many equipment models are available for this purpose. Due to possible device sensitivity and reproducibility differences during examination, the main goal of this study was to compare electrocardiographic parameters of dogs using two different C-EKG systems: Wincardio Micromed ® (WIN) and TEB ECGPC ® (TEB). Forty two healthy male and female dogs of different breeds (Cocker Spaniel, Dachshund, Labrador, Pinscher, Pitbull Terrier, Poodle, Schnauzer, Shih Tzu, Yorkshire and mongrel dogs), with age between 4 months and 16 years old were grouped according to weight and evaluated by both systems. The electrocardiographic measurements were performed on DII lead for both systems. The study showed that the TEB system was more sensitive for measurement of P wave and QRS complex duration, while the WIN system showed more sensitivity for the measurements of amplitude of the same parameters. The larger animals (26-37kg) showed greater variance in the measurements of P wave and QRS complex amplitude and duration than the groups of medium (14-25kg) or smaller (3-13kg) dogs. These differences must be considered when using diverse computerized electrocardiography systems to perform measurements due to the possibility of erratic interpretation of the results between veterinary medicine services.

Research paper thumbnail of Comparison between splenic and lymph node aspirations as sampling methods for the parasitological detection of Leishmania chagasi infection in dogs

Memórias do Instituto Oswaldo Cruz, 2004

The sensitivities of spleen and lymph node cultures for the diagnosis of canine visceral leishman... more The sensitivities of spleen and lymph node cultures for the diagnosis of canine visceral leishmaniasis were compared in 64 anti-Leishmania antibody positive dogs from an endemic area in Brazil. The sensitivity of spleen cultures for Leishmania detection was 97.9%; in lymph node cultures it was 25%. Positive spleen culture was more frequent (p = 0.048, Fisher's exact probability test) in symptomatic (28 out of 33 animals) than in asymptomatic animals (19 out of 31 animals). These results support the use of spleen instead of lymph node aspiration as the choice method for the parasitological diagnosis of the infection.

Research paper thumbnail of Enhancement of Experimental Cutaneous Leishmaniasis by Leishmania Molecules Is Dependent on Interleukin-4, Serine Protease/Esterase Activity, and Parasite and Host Genetic Backgrounds

Infection and Immunity, 2011

Most inbred strains of mice, like the BALB/c strain, are susceptible to Leishmania amazonensis in... more Most inbred strains of mice, like the BALB/c strain, are susceptible to Leishmania amazonensis infections and resistant to Leishmania braziliensis infections. This parasite-related difference could result from the activity of an L. amazonensis-specific virulence factor. In agreement with this hypothesis, it is shown here that the intravenous injection of BALB/c mice with L. amazonensis amastigote extract (LaE) but not the L. braziliensis extract confers susceptibility to L. braziliensis infection. This effect was associated with high circulating levels of IgG1 anti-L. amazonensis antibodies and with an increase in interleukin-4 (IL-4) production and a decrease in gamma interferon production by draining lymph node cells. Moreover, the effect was absent in IL-4knockout mice. The biological activity in the LaE was not mediated by amphiphilic molecules and was inhibited by pretreatment of the extract with irreversible serine protease inhibitors. These findings indicate that the LaE contains a virulence-related factor that (i) enhances the Leishmania infection by promoting Th2-type immune responses, (ii) is not one of the immunomodulatory Leishmania molecules described so far, and (iii) is either a serine protease or has an effect that depends on that protease activity. In addition to being Leishmania species specific, the infection-enhancing activity was also shown to depend on the host genetic makeup, as LaE injections did not affect the susceptibility of C57BL/6 mice to L. braziliensis infection. The identification of Leishmania molecules with infection-enhancing activity could be important for the development of a vaccine, since the up-or downmodulation of the immune response against a virulence factor could well contribute to controlling the infection.

Research paper thumbnail of Production of Monoclonal Antibodies Against Canine Leukocytes

Hybridoma and Hybridomics, 2004

A panel of anti-canine leukocyte monoclonal antibodies (MAbs) was produced by immunizing BALB/c m... more A panel of anti-canine leukocyte monoclonal antibodies (MAbs) was produced by immunizing BALB/c mice with canine peripheral blood mononuclear cells (PBMC), either resting or stimulated with concanavalin A (ConA). Three out of 28 clones—IH1, AB6, and HG6–screened by ...

Research paper thumbnail of A Novel Monoclonal Antibody Against Canine Monocytes/Macrophages

Hybridoma and Hybridomics, 2004

The production and partial characterization of a monoclonal antibody, the IgG 1 IH1, which recogn... more The production and partial characterization of a monoclonal antibody, the IgG 1 IH1, which recognizes an antigen distributed in canine monocytes/macrophages, is reported here. The distribution and apparent molecular weight of the antigen recognized by the IH1 MAb was determined in peripheral blood leukocytes, peripheral blood monocyte-derived macrophages and tissue sections of spleen, liver and skin, using Western blotting, immunocytochemistry, immunohistochemistry and flow cytometry. The IH1 MAb-recognized antigen was detected in Western blotting under non-reducing conditions spread out as a large band covering the position corresponding to the migration of molecules with molecular weights from 55 to 73 kDa. The IH1 MAb labeled blood monocytes, tissue macrophages in lymph nodes, and in the mantle zone of the spleen, and Kupffer cells in the liver. It did not react with human cells. In flow cytometric analysis, the IH1 MAb reacted with a subpopulation of monocytes. The MAb described herein may become a valuable tool for diagnosis and research on canine diseases.

Research paper thumbnail of Ectopia ureteral unilateral congênita em uma cadela Teckel Dachshund com pelagem arlequim: relato de caso

Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 2012

O presente relato apresenta a primeira descrição de ectopia ureteral congênita na raça Teckel Dac... more O presente relato apresenta a primeira descrição de ectopia ureteral congênita na raça Teckel Dachshund, diagnosticada em uma cadela com pelagem arlequim. O animal, aos dois meses de idade, apresentava sinais de incontinência urinária e cistite bacteriana, sendo submetido a um plano diagnóstico para confirmação de ureter ectópico. A urografia excretora revelou hidroureter direito com desembocadura caudal ao trígono da bexiga. O exame físico do animal evidenciou ainda hérnia inguinal bilateral, o que reforçou a caracterização da origem congênita das alterações. Aspectos de bem-estar animal são também discutidos.

Research paper thumbnail of An Assessment of the Genetic Diversity of Leishmania infantum Isolates from Infected Dogs in Brazil

American Journal of Tropical Medicine and Hygiene, 2012

Correlations between the genetic diversity of Leishmania infantum (syn. L. chagasi)isolatesandthe... more Correlations between the genetic diversity of Leishmania infantum (syn. L. chagasi)isolatesandtheirrespective geographic origins support the theoretic assumption that visceral leishmaniasis probably originated in the Old World. Because dogs are widely considered to be the main reservoir of this disease, the present study aimed to investigate the degree of genetic divergence among 44 leishmanial canine isolates from two Brazilian cities, Jequié and Campo Grande, located approximately 2,028 km from each other. We hypothesized that a low degree of genetic divergence would be observed among these isolates. In fact, statistical analyses found no significant differences between the isolates using both random amplified polymorphic DNA and multilocus microsatellite typing genotyping techniques with three and seven markers, respectively. These findings provide support for the recent introduction of L. infantum into the New World.

Research paper thumbnail of Evaluating the Accuracy of Molecular Diagnostic Testing for Canine Visceral Leishmaniasis Using Latent Class Analysis

PLoS ONE, 2014

Host tissues affected by Leishmania infantum have differing degrees of parasitism. Previously, th... more Host tissues affected by Leishmania infantum have differing degrees of parasitism. Previously, the use of different biological tissues to detect L. infantum DNA in dogs has provided variable results. The present study was conducted to evaluate the accuracy of molecular diagnostic testing (qPCR) in dogs from an endemic area for canine visceral leishmaniasis (CVL) by determining which tissue type provided the highest rate of parasite DNA detection. Fifty-one symptomatic dogs were tested for CVL using serological, parasitological and molecular methods. Latent class analysis (LCA) was performed for accuracy evaluation of these methods. qPCR detected parasite DNA in 100% of these animals from at least one of the following tissues: splenic and bone marrow aspirates, lymph node and skin fragments, blood and conjunctival swabs. Using latent variable as gold standard, the qPCR achieved a sensitivity of 95.8% (CI 90.4-100) in splenic aspirate; 79.2% (CI 68-90.3) in lymph nodes; 77.3% (CI 64.5-90.1) in skin; 75% (CI 63.1-86.9) in blood; 50% (CI 30-70) in bone marrow; 37.5% (CI 24.2-50.8) in left-eye; and 29.2% ) in right-eye conjunctival swabs. The accuracy of qPCR using splenic aspirates was further evaluated in a random larger sample (n = 800), collected from dogs during a prevalence study. The specificity achieved by qPCR was 76.7% (CI 73.7-79.6) for splenic aspirates obtained from the greater sample. The sensitivity accomplished by this technique was 95% (CI 93.5-96.5) that was higher than those obtained for the other diagnostic tests and was similar to that observed in the smaller sampling study. This confirms that the splenic aspirate is the most effective type of tissue for detecting L. infantum infection. Additionally, we demonstrated that LCA could be used to generate a suitable gold standard for comparative CVL testing.