Asymptomatic dogs are highly competent to transmit Leishmania (Leishmania) infantum chagasi to the natural vector

We evaluated the ability of dogs naturally infected with Leishmania (Leishmania) infantum chagasi to transfer the parasite to the vector and the factors associated with transmission. Thirty-eight infected dogs were confirmed to be infected by direct observation of Leishmania in lymph node smears. Dogs were grouped according to external clinical signs and laboratory data into symptomatic (n= 24) and asymptomatic (n= 14) animals. All dogs were sedated and submitted to xenodiagnosis with F1-laboratory-reared Lutzomyia longipalpis. After blood digestion, sand flies were dissected and examined for the presence of promastigotes. Following canine euthanasia, fragments of skin, lymph nodes, and spleen were collected and processed using immunohistochemistry to evaluate tissue parasitism. Specific antibodies were detected using an enzyme-linked immunosorbent assay. Antibody levels were found to be higher in symptomatic dogs compared to asymptomatic dogs (p= 0.0396). Both groups presented amastigotes in lymph nodes, while skin parasitism was observed in only 58.3% of symptomatic and in 35.7% of asymptomatic dogs. Parasites were visualized in the spleens of 66.7% and 71.4% of symptomatic and asymptomatic dogs, respectively. Parasite load varied from mild to intense, and was not significantly different between groups. All asymptomatic dogs except for one (93%) were competent to transmit Leishmania to the vector, including eight (61.5%) without skin parasitism. Sixteen symptomatic animals (67%) infected sand flies; six (37.5%) showed no amastigotes in the skin. Skin parasitism was not crucial for the ability to infect Lutzomyia longipalpis but the presence of Leishmania in lymph nodes was significantly related to a positive xenodiagnosis. Additionally, a higher proportion of infected vectors that fed on asymptomatic dogs was observed (p= 0.0494). Clinical severity was inversely correlated with the infection rate of sand flies (p= 0.027) and was directly correlated with antibody levels (p= 0.0379). Age and gender did not influence the transmissibility. Our data show that asymptomatic dogs are highly infective and competent for establishing sand fly infection, indicating their role in maintaining L. (L.) infantum chagasi cycle as well as their involvement in VL spreading in endemic areas. © 2013 Elsevier B.V.

Leishmaniasis causes oxidative stress and alteration of oxidative metabolism and viability of neutrophils in dogs

The aim of the present study was to test the hypothesis that oxidative stress and alteration of oxidative metabolism and apoptosis of neutrophils in dogs vary with the stage of leishmaniasis and to determine the contribution of uremia to such alterations. Dogs with leishmaniasis were classified into two stages: moderate (Leish II, n = 20) or very severe (i.e. with concurrent uremia; Leish IV, n = 20) according to the LeishVet Consensus. The two leishmaniasis groups were compared with uremic dogs without leishmaniasis (Uremic, n = 10) and to healthy dogs (Control, n = 30). To determine oxidative stress, total antioxidant/oxidant capacity, lipid peroxidation, total glutathione and the plasma antioxidants albumin, uric acid and bilirubin were quantified. Superoxide production was determined using the hydroethidine probe and viability and apoptosis were measured using annexin V-PE by capillary flow cytometry. Oxidative stress was present in both uremia and leishmaniasis with reduced total antioxidant capacity and was associated with increased induced production of superoxide and apoptosis. The greatest amount of oxidants was observed in animals with moderate disease only. Neutrophils from uremic dogs with and without leishmaniasis had decreased viability and an increased apoptosis rate in addition to increased lipid peroxidation. In conclusion, oxidative stress occurs in both stages of leishmaniasis with differences in intensity and levels of plasma markers; however, uremia does contribute to the decreased spontaneous viability of neutrophils in dogs in the final stage of the disease. © 2013 Elsevier Ltd. All rights reserved.

CD95 (FAS) and CD178 (FASL) induce the apoptosis of CD4+ and CD8+ cells isolated from the peripheral blood and spleen of dogs naturally infected with Leishmania spp

Infected dogs are urban reservoirs of Leishmania chagasi, which is a causative agent of visceral leishmaniasis (VL). Dogs exhibit immune suppression during the course of this disease, and lymphocyte apoptosis is involved in this process. To investigate apoptosis and the expression levels of FAS-FAS-associated death domain protein (CD95 or APO-1), FASL-FAS ligand protein (CD178), and TRAIL-TNF-related apoptosis-inducing ligand (CD253) receptors in peripheral blood mononuclear cells and spleen leukocytes from 38 symptomatic dogs with moderate VL and 25 healthy dogs were evaluated by flow cytometry. The apoptosis rate of blood and splenic CD4+ and CD8+ cells was higher in infected dogs than in healthy dogs. The expression levels of FAS and FASL in blood and splenic CD4+ cells were lower in infected dogs than in healthy dogs. FAS expression in CD8+ cells was higher in infected dogs than in healthy dogs; in contrast, FASL expression was lower in infected dogs. The expression of the TRAIL receptor increased only in splenic CD8+ cells from infected dogs. The FAS and FAS-L blocking antibodies confirmed the importance of these receptors in apoptosis. Our results enhance the current understanding of the immune response in dogs infected with L. chagasi, facilitating the future development of therapeutic interventions to reduce lymphocyte depletion. © 2013 Elsevier B.V.

Longitudinal analysis of serological tests officially adopted by the Brazilian Ministry of Health for the diagnosis of canine visceral leishmaniasis in dogs vaccinated with Leishmune®

Development of vaccines against canine visceral leishmaniasis (CVL) may provide a prophylactic barrier, but antibody response detected by standard diagnostic techniques may not separate vaccinated from naturally infected dogs. Moreover, anti-Leishmania antibody levels in vaccinated dogs may be detectable for months. Accordingly, the aim of the present study was to comparatively evaluate an in-house ELISA with three serological tests officially adopted by the Brazilian Ministry of Health for the diagnosis of CVL in dogs vaccinated with Leishmune®. A total of 18 mongrel dogs were submitted to a complete protocol of the vaccine, monitored and evaluated in 5 times (T0-T4) up to 180 days after T0. Twenty-one days after the first dose (T1), 50% of the dogs were seropositive by the in-house ELISA and 5.5% by IFAT, while by the official ELISA and DPP® CVL rapid test all dogs tested negative. At time T2, 42 days after of the first dose, 100%, 83.3%, 11.1%, and 5.5% of the dogs were seropositive by the in-house ELISA, IFAT, official ELISA kit and the DPP® CVL rapid test, respectively. Ninety days after the first dose (T3), 100%, 83.3%, 72.2% and 33.3% of the dogs were seropositive by the in-house ELISA, official ELISA kit, IFAT, and the DPP® CVL rapid test, respectively. Finally, at time T4, 88.8%, 33.3%, 11.1% and 5.5% of the dogs were seropositive by the in-house ELISA, official ELISA kit, DPP® CVL rapid test and IFAT, respectively. In conclusion, dogs vaccinated with Leishmune® cross-react by an in-house ELISA and by the three official Brazilian serological tests for the diagnosis of canine visceral leishmaniasis up to six months after the first vaccine dose, and may be mistakenly diagnosed and removed. © 2013 Elsevier B.V.

Stocking density of Nile tilapia in cages placed in a hydroelectric reservoir

The Brazilian government has been encouraging fish farming in cages in federal water bodies, including hydroelectric reservoirs. Despite the government support, it is a new activity and the production model still needs some adjustment to reduce the production costs and achieve sustainability. The aims of this study were to determine the appropriate stocking density of Nile tilapia in cages in a hydroelectric reservoir and to evaluate to what extent fish size selection could improve their uniformity. Twelve cages (6m3) were placed at the Fish Farmers' Cooperative of Santa Fé do Sul and Region, Ilha Solteira reservoir, São Paulo, Brazil (20°12'10″S, 50°58'31.15″W). In stage I (initial fish weight, 78g), four stocking densities were tested: D1-800, D2-2000, D3-2500 and D4-3000 fish/cage, with three replicates. At the end of this stage (average fish weight, 255g), the fish were selected into three sizes, except for D1. In stage II, four stocking densities were tested, designed to obtain the following final production: D1-100kg/m3 (800 non-selected fish/cage), D2-80kg/m3 (600 fish/cage), D3-100kg/m3 (800 fish/cage) and D4-120kg/m3 (900 fish/cage). The trial ended when the fish weighed 800g. By reducing the initial stocking density from 2500 to 800 tilapia juveniles per cage, there was no need for selection. The growth performance was higher, the feed conversion rate was better and the time taken to reach harvesting was shorter. Consequently, the production cost reduced and the operating profit increased. Using the lowest initial stocking density, the risk of disease outbreak was also lower, and there was no need to use drugs for disease control since the mortality rate and occurrences of disease and deformity decreased and the dissolved oxygen level inside the cages was higher. © 2013 Elsevier B.V.

Effects of cold fronts on MODIS-derived sensible and latent heat fluxes in Itumbiara reservoir (Central Brazil)

In this work we investigate the cold front passage effects on sensible and latent heat flux in a tropical hydroelectric reservoir. The study area, Itumbiara reservoir (Goiás State/Brazil) at the beginning of the austral winter, is characterized by the presence of a weak thermal stratification and the passage of several cold fronts from higher latitudes of South America. Sensible and latent heat fluxes were estimated considering the atmospheric boundary layer stability. In situ and MODIS water surface temperature data were used to adjust the coefficients for momentum and heat exchanges between water and atmosphere and spatialize the sensible and latent heat fluxes. The results showed that during a cold front event the sensible heat flux can be up to five times greater than the flux observed before. The latent heat flux tends to decrease during the cold front but increase again after the passage. The highest values of heat loss were observed at littoral zone and some Reservoir's embayment. The heat loss intensification can be separated in two moments: first, during the cold front passage, when the wind speed increases and the air temperature decreases; second, after the cold front passage, with air humidity decreasing. This can be considered a key process to understanding the heat loss in the Itumbiara reservoir. © 2013 COSPAR. Published by Elsevier Ltd. All rights reserved.

Interference of cage fish farm on diet, condition factor and numeric abundance on wild fish in a Neotropical reservoir

Historically, the ichthyofauna of large Brazilian rivers has been subject to anthropogenic interference, such as impoundments. Currently, cage fish farming systems are a new source of impact on aquatic ecosystems. The objective of this study was to characterise the impact of freshwater fish farms on the feeding of five species of Neotropical freshwater fish. Specimens of Astyanax altiparanae, Galeocharax knerii, Iheringicthys labrosus, Pimelodus maculatus and Plagioscion squamosissimus were sampled in areas around two systems of cage fish farming (CF), and two control areas (CT) that were not influenced by this activity. Results show that there were significant changes in the diet of trophic generalist species (A. altiparanae, P. maculatus and I. labrosus) accompanied by a related increase in the condition factor values of these species in cage areas. Trophic specialist species, such as the carnivorous fish species G. knerii and P. squamosissimus, presented small differences between the CF and CT areas with regard to diet and showed no differences in other analyses performed. In conclusion, cage fish farms can affect the natural diet of trophic generalist fish species, directly affecting the nutritional status (condition factor), where food wastes was found to be one of the principal items consumed by this trophic guild. Results indicate that these species are responsible for recycling a great quantity of organic matter transferred by this type of activity, which, along with local fishery activities, contribute to mitigation of associated processes of eutrophication. © 2013 Elsevier B.V.

Leishmaniose visceral canina: aspectos clínico-laboratoriais, histopatologia renal e testes específicos para diagnóstico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização antigênica e genotípica de isolados do vírus rábico, de quirópteros da cidade de Botucatu-SP e região

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação diagnóstica para Leishmania spp. e Trypanosoma cruzi em gatos domésticos procedentes da associação protetora dos animais do município de Ilha Solteira, SP, Brasil

Pós-graduação em Doenças Tropicais - FMB

Pesquisa de mutação do gene predito da lectina ligante de manose em cães: comparação entre dois alvos para diagnóstico molecular de leishmaniose visceral canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão das quimiocinas MIP-1α e MCP-1 em relação à carga parasitária em cães com leishmaniose visceral

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão em Escherichia coli de uma proteína recombinante do gene A2 de Leishmania chagasi e utilização desta proteína no diagnóstico da leishmaniose visceral canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise de cepas de Escherichia coli Shiga toxigênica (STEC) e E. coli patogênica extraintestinal (ExPEC) isoladas de cachorros diarréicos atendidos em clínica privada no município de Ituverava, SP

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise da composição celular e imunodetecção de MIF em linfonodos de cães com Leishmaniose Visceral

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)