PREVALENCE OF AMERICAN TRYPANOSOMIASIS AND LEISHMANIASES IN DOMESTIC DOGS IN A RURAL AREA OF THE MUNICIPALITY OF SÃO JOÃO DO PIAUÍ, PIAUÍ STATE, BRAZIL

SUMMARY Chagas disease and the leishmaniases are endemic zoonoses of great importance to public health in the state of Piauí, Brazil. The domestic dog (Canis familiaris) is a major reservoir, host of Trypanosoma cruzi and Leishmania spp. in both urban and rural areas, playing an important role in the transmission of these parasites. The present study evaluated the prevalence of both infectious diseases in dogs of a rural area in the municipality of São João do Piauí, Piauí State. One hundred twenty-nine blood samples were collected for serological assessment: for the leishmaniases, 49 (38%) animals tested positive by the Dual-Path Platform technology (DPP), nine (6%) by the Enzyme-linked Immunosorbent Assay (ELISA), and 19 (14.7%) by the Indirect Fluorescent Antibody test (IFA); while for American Trypanosomiasis, 36 (28%) dogs were reagent by ELISA and 21 by IFA. Of the 129 dogs sampled, 76 were submitted to xenodiagnosis, bone marrow aspiration and skin biopsy to perform parasitological tests whose results showed only one (2.3%) positive skin sample for Trypanosoma caninum and one positive xenodiagnosis for T. cruzi, both results confirmed by molecular assays. Three hundred triatomines of the species Triatoma brasiliensis and 552 phlebotomines - 509 (97%) of the species Lutzomyia longipalpis, were also captured

Vacinas anti-leishmaniose visceral canina podem interferir no diagnóstico sorológico preconizado pelo Ministério da Saúde brasileiro?

Submitted by Sandra Infurna ([email protected]) on 2017-07-05T13:29:13Z No. of bitstreams: 1 paula_deluca_etal_IOC_2017,pdf.pdf: 448643 bytes, checksum: bbc723df2f49f3a95ff3262d16e7568a (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2017-07-05T13:43:05Z (GMT) No. of bitstreams: 1 paula_deluca_etal_IOC_2017,pdf.pdf: 448643 bytes, checksum: bbc723df2f49f3a95ff3262d16e7568a (MD5)Made available in DSpace on 2017-07-05T13:43:05Z (GMT). No. of bitstreams: 1 paula_deluca_etal_IOC_2017,pdf.pdf: 448643 bytes, checksum: bbc723df2f49f3a95ff3262d16e7568a (MD5) Previous issue date: 2017Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Exército Brasileiro. 1. Batalhão de Guardas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.O objetivo deste trabalho foi avaliar a soroconversão em cães imunizados com as vacinas Leishmune® e Leish tec®, através do teste imunocromatográfico rápido DPP® (Dual Path Platform) (DPP LVC) e do ensaio imunoenzimático (EIE) durante um ano após o protocolo vacinal. Trata-se de um estudo onde 28 cães divididos em dois grupos foram imunizados cada um com uma vacina anti - LVC e acompanhados durante um ano através de avaliação clínica e exames laboratoriais. Foi possível acompanhar 22 (78.5%) cães. Nos exames dos tempos 1, 2 e 3 (respectivamente 30 dias, 6 messes e 1 ano após a vacinação) os resultados de todos os cães também foram negativos para LVC, exceto de um cão que recebeu a vacina Leish tec® e soroconverteu no DPP LVC no T2, após 6 meses a vacina. Os exames posteriores deste cão foram negativos. Os resultados do presente estudo demostraram que, em área não endêmica e mesmo em diferentes tempos de avaliação, cães vacinados contra LVC, independente da vacina utilizada, não foram capazes de soroconverter no protocolo utilizado pelo Ministério da Saúde brasileiro (DPP/EIE).The objective of the current research was to assess seroconversion in dogs immunized with Leishmune® and Leish Tec® vaccines using rapid chromatographic immunoassay DPP® (Dual Path Platform) (DPP CVL) and enzyme immunoassay (EIE) up to one year after the vaccination protocol. The study sample comprised 28 dogs divided into two groups, each group immunized with an anti-CVL vaccine and clinically monitored for one year through clinical evaluation and laboratory tests. 22 (78.5%) dog were monitored. During the evaluation time (T1-30 days, T2-6 months, and T3-1 year after vaccination) the results for all dogs were negative for CVL, except for one animal vaccinated with Leish tec® that seroconverted in the DPP CVL test at T2. Subsequent examinations of this dog were negative. Our results showed that in a non-endemic area, even at different evaluation times, dogs vaccinated against CVL with Leishmune® or Leish tec® did not seroconvert in the serological protocol used by the Brazilian Ministry of Health (DPP/EIE)

Frequency of detection and load of amastigotes in the pancreas of Leishmania infantum-seropositive dogs: clinical signs and histological changes

Abstract Background Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and is highly lethal in humans and dogs if left untreated. The frequency of this parasite and associated histological changes in the pancreas of dogs are poorly studied. Therefore, the objectives of this study were to evaluate the frequency of detection and load of amastigotes in the pancreas of L. infantum-seropositive dogs and to identify the clinical signs and histological changes associated with parasitism of this organ. Methods One hundred forty-three dogs from an endemic area in Brazil that tested seropositive for L. infantum were studied. The dogs were clinically examined, killed, and necropsied between 2013 and 2014. One fragment of the pancreas was randomly collected for histopathology and immunohistochemistry, and spleen and bone marrow were collected for culture. Results Leishmania amastigotes were detected in the pancreas of 22 dogs (15.4%) by immunohistochemistry, all exhibiting L. infantum parasitism in the spleen and/or bone marrow. Poor body condition and cachexia were only associated with infection of the pancreas with Leishmania spp. (p = 0.021) and were found in 40.9% of dogs with pancreatic infection. Anorexia, vomiting, and/or diarrhea were observed in 9.2% of dogs with pancreatitis. The median parasite load in the pancreas was 1.4 infected macrophages/mm2. Pancreatic histological changes and their frequencies were: granulomatous pancreatitis (28.0%), lymphoplasmacytic pancreatitis (23.8%), acinar cell degeneration (6.3%), fibrosis (5.6%), hemorrhage (2.1%), eosinophilic pancreatitis (0.7%), suppurative pancreatitis (0.7%), and necrosis (0.7%). Conclusions The present results demonstrate that L. infantum is one of the etiological agents of chronic pancreatitis in dogs; however, the frequency of detection and parasite load are low in this organ. The lack of an association of poor body condition and cachexia with pancreatitis and the low frequency of clinical signs commonly associated with pancreatitis suggest that a significant portion of the organ is not affected by this parasite. On the other hand, the association of poor body condition and cachexia with concomitant infection of the pancreas, spleen, and/or bone marrow with this parasite suggests that these manifestations are the result of a more advanced stage of canine visceral leishmaniasis. Graphic abstrac

Occurrence of Leishmania infantum in the central nervous system of naturally infected dogs: Parasite load, viability, co-infections and histological alterations

Submitted by Sandra Infurna ([email protected]) on 2017-11-16T11:11:07Z No. of bitstreams: 1 maria_amendoeira_etal_IOC_2017.pdf: 3406658 bytes, checksum: b56818c4923ad9b09eb9ca36b7069392 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2017-11-16T11:25:00Z (GMT) No. of bitstreams: 1 maria_amendoeira_etal_IOC_2017.pdf: 3406658 bytes, checksum: b56818c4923ad9b09eb9ca36b7069392 (MD5)Made available in DSpace on 2017-11-16T11:25:00Z (GMT). No. of bitstreams: 1 maria_amendoeira_etal_IOC_2017.pdf: 3406658 bytes, checksum: b56818c4923ad9b09eb9ca36b7069392 (MD5) Previous issue date: 2017Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Vigilância em Leishmanioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Serviço de Anatomia Patológica. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Serviço de Anatomia Patológica. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Epidemiologia Clínica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxoplasmose. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evando Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ. Brasil.Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and little is known about the occurrence and pathogenesis of this parasite in the CNS. The aims of this study were to evaluate the occurrence, viability and load of L. infantum in the CNS, and to identify the neurological histological alterations associated with this protozoan and its co-infections in naturally infected dogs. Forty-eight Leishmania-seropositive dogs from which L. infantum was isolated after necropsy were examined. Cerebrospinal fluid (CSF) samples were analyzed by parasitological culture, quantitative real-time PCR (qPCR) and the rapid immunochromatographic Dual Path Platform test. Brain, spinal cord and spleen samples were submitted to parasitological culture, qPCR, and histological techniques. Additionally, anti-Toxoplasma gondii and anti-Ehrlichia canis antibodies in serum and distemper virus antigens in CSF were investigated. None of the dogs showed neurological signs. All dogs tested positive for L. infantum in the CNS. Viable forms of L. infantum were isolated from CSF, brain and spinal cord in 25% of the dogs. Anti-L. infantum antibodies were detected in CSF in 61% of 36 dogs. Inflammatory histological alterations were observed in the CNS of 31% of the animals; of these, 66% were seropositive for E. canis and/or T. gondii. Amastigote forms were associated with granulomatous non-suppurative encephalomyelitis in a dog without evidence of co-infections. The highest frequency of L. infantum DNA was observed in the brain (98%), followed by the spinal cord (96%), spleen (95%), and CSF (50%). The highest L. infantum load in CNS was found in the spinal cord. These results demonstrate that L. infantum can cross the blood-brain barrier, spread through CSF, and cause active infection in the entire CNS of dogs. Additionally, L. infantum can cause inflammation in the CNS that can lead to neurological signs with progression of the disease

<i>Leishmania infantum</i> parasite load expressed as the natural logarithm of the number of parasite genome equivalents (gEq)/ng in the brain and spinal cord of 14 dogs with inflammatory alterations in the CNS according to serum positivity for anti-<i>E</i>. <i>canis</i> and anti-<i>T</i>. <i>gondii</i> antibodies.

<p>The vertical black lines indicate the median parasite load. The horizontal dotted lines indicate the interquartile range. The blue dots represent the parasite load of each dog.</p

<i>Leishmania infantum</i> parasite load expressed as the natural logarithm of the number of parasite genome equivalents (gEq)/ng in the brain and spinal cord of naturally infected dogs with and without histological alterations in the central nervous system.

<p>The horizontal black lines indicate the median parasite load. The vertical dotted lines indicate the interquartile range. The horizontal dotted lines indicate the lower limit of positivity (threshold). The blue dots represent the parasite load of each dog. The empty dots are the outliers of parasite load defined by the boxplot.</p

Frequency of <i>Leishmania</i> positivity in the central nervous system and spleen of naturally infected dogs detected by histopathology (HP), immunohistochemistry (IHC), <i>in situ</i> hybridization (ISH), parasitological culture (PC), DPP^® assay, and qualitative real-time PCR (qPCR).

<p>* 44 samples were investigated by qPCR; ** 36 samples were investigated by qPCR and DPP^® assay. CSF = cerebrospinal fluid.</p

<i>Leishmania infantum</i> parasite load expressed as the natural logarithm of the number of parasite genome equivalents (gEq)/ng in the central nervous system (CNS) and spleen of naturally infected dogs according to a positive or negative <i>L</i>. <i>infantum</i> result in the CNS by the parasitological techniques (culture and histological techniques).

<p>The horizontal black lines indicate the median parasite load. The vertical dotted lines indicate the interquartile range. The horizontal dotted lines indicate the lower limit of positivity (threshold). The blue dots represent the parasite load of each dog. The empty dots are the outliers of parasite load defined by the boxplot. CSF = cerebrospinal fluid. CNS = central nervous system.</p