José Eduardo Tolezano

Ecoepidemiological aspects of american cutaneous leishmaniasis in the state of São Paulo, Brazil

The aim of this study is to review some of the ecoepidemiological aspects of american cutaneous leishmaniasis (ACL) in the State of São Paulo, Brazil. During the first half of this century ACL occurred in São Paulo, predominantly on the bank of the Tietê River, where there were railroad constructions and there was inevitable contact between workers and forested areas. Mans activities resulted in a drastic reduction of the forested regions of the State and molded the present landscape found in São Paulo, which brought a gradual change in the epidemiology of ACL during this century. Currently ACL can be considered as an endemic disease. Nowadays, ACL is found in different regions of São Paulo State, and is no longer limited to the bank of the Tietê River. The disease occurs in all age groups and sexes. Lack of knowledge about wild reservoir hosts of Leishmania (V.) braziliensis has simulated speculation about the possible role played by domestic animals (dogs and equines). Mans activities also favoured Lutzomyia intermedia a sandfly species which can clearly thrive in changed environments. L.(V.) braziliensis continues to be transmitted, even after decades of forest destruction in São Paulo.

Tegumentary Leishmaniasis as a Manifestation of Immune Reconstitution Inflammatory Syndrome in 2 Patients with AIDS

Immune reconstitution inflammatory syndromes (IRISs) have been reported in patients with human immunodeficiency virus/acquired immunodeficiency syndrome (AIDS) since the introduction of highly active antiretroviral therapy (HAART). This syndrome is characterized by clinical manifestations of opportunistic infections when signs of immune reconstitution are observed during therapy. We report on leishmaniasis, suggestive of HAART-induced IRIS, in 2 patients with AIDS. After beginning HAART, 1 patient presented with disseminated, tegumentary lesions, whereas the other patients preexisting lesions worsened and became more extensive; however, at the same time, their CD4(+) T cell counts were recovering and their virus loads were decreasing significantly. The lesions healed with anti-Leishmania therapy.

Regional variation in toxoplasmosis seronegativity in the São Paulo metropolitan region

Toxoplasmosis is a highly prevalent zoonotic human infection caused by the Apicomplexa protozoon Toxoplasma gondii. The acute disease is usually mild or asymptomatic, except for foetal infection transmitted by acutely infected pregnant women, which courses as a devastating disease. In order to determine possible regional variations in risk factors, we studied the frequency of seronegativity in areas of the São Paulo Metropolitan Region, comparing titers and age groups. The prevalence of seronegativity was determined retrospectively in 1286 pregnant women receiving prenatal care at public health services in four selected areas of the São Paulo Metropolitan Region of similar socioeconomic background. The São Paulo City area had the higher frequency of seronegativity (41.1%), followed by the Northwest (31.5%) and Southwest (29.9%) areas, with similar intermediate levels, and by the Northeast (22.5%) area with the lowest frequency (p < 0.001). A rough estimate disclosed about 280 infected infants/year in the São Paulo Metropolitan Region. Serological titers analyzed by age group suggested a decline in antibody levels with age, as shown by a lower frequency of higher titers in older groups. Our study emphasizes the importance of determining the regional prevalence of toxoplasmosis for proper planning of public health prenatal care.

Investigação epidemiológica de, um novo caso de leishmaniose visceral ocorrido na Grande São Paulo, Brasil

Descreve-se uma investigacao epidemiologica realizada em zona urbana do municipio de Sao Paulo, Brasil, para esclarecer um caso de leishmaniose visceral ocorrido em crianca de 2 anos de idade, nascida e sempre residente no local. Afastou-se a possibilidade de transmissao por via transfusional e por vetor biologico, tendo como base os dados levantados da anamnese do doente, os resultados de inqueritos realizados na area em populacao humana, utilizando testes de imunofluorescencia indireta, hemaglutinacao passiva e intradermoreacao de Montenegro, em populacao canina com o teste de imunofluorescencia indireta, alem de pesquisa entomologica em mata residual.

Chave de identificação ilustrada dos Phlebotominae(Diptera, Psychodidae) do Estado de São Paulo

Flebotomineos sao insetos de importância medica, responsaveis pela transmissao de parasitas do genero Leishmania entre humanos e animais nao humanos, os quais sao encontrados no estado de Sao Paulo, Brasil. A correta identificacao das especies envolvidas na transmissao de doencas e fundamental para elaboracao de estrategias para o controle desses vetores. Para produzir uma chave ilustrada para as 67 especies de machos e 57 especies de femeas de flebotomineos registradas no estado de Sao Paulo, examinamos e ilustramos especimes montados em lâminas de microscopia obtidos de colecoes entomologicas.

Value of the oral swab for the molecular diagnosis of dogs in different stages of infection with Leishmania infantum

This study was based on the need to employ a sensitive and specific method with samples that could be easily collected for diagnosing dogs infected with Leishmania infantum. To this end, we used real time-PCR (qPCR) to assess the value of the oral swab (OS) in detecting infected sick dogs (SD; n=62), including, for the first time, the analysis of apparently healthy infected dogs (AD; n=30), both from endemic areas for visceral leishmaniasis (VL). For comparison, we also evaluated the performance of the conjunctival swab (CS), blood (BL), lymph node (LN) and serology. We detected the presence of Leishmania DNA in the oral cavity in 62 out of the 92 dogs studied. The OS positivity (67.4%) was equivalent to the CS (68.5%) (p>0.05), higher than BL (52.2%) (p≤0.05), and lower than LN (84.8%) (p≤0.05). OS and CS performed well in SD dogs (82.3% and 83.9%, respectively) but not in AD dogs (36.7% for both samples). BL showed the lowest positivity (52.2%) and provided equivalent results between AD (60.0%) and SD (48.4%) dogs (p>0.05). LN yielded the highest positivity (84.8%), and it was also higher in the SD population (93.5%) compared to the AD population (66.7%) (p≤0.05). Parasite load was high in LN, moderate in OS and CS, and low in BL, showing the relationship between the levels of parasitism and the positivity rates found in these samples. Serology was positive in 82.2% of the SD group and in 70% of the AD dogs (p>0.05). Among the 20 seronegative dogs, seven (35%) were positive in either OS or CS, and 12 (60%) were positive when both noninvasive samples were jointly considered. The OS/CS combination resulted in a significant increase of positivity (p≤0.05) for the AD dogs (from 36.7% to 63.4%), as well as OS/serology (80%) and OS/CS/serology (83.4%). For the SD population, positivity reached up to 95.2% with the same combinations, showing that combination of samples and/or tests is required for the identification of dogs infected with L. infantum and that the OS and CS combination based on qPCR notably improves the detection of both AD and SD dogs. In conclusion, OS proved to be a suitable sample for the molecular diagnosis of infected dogs with clinical signs of VL, but not for dogs with inapparent infection. For these, we recommend the combination of OS results with CS and/or serology in order to reach relevant positivity for L. infantum. Finally, another advantage of using OS or both noninvasive samples is the increased likelihood of diagnosing seronegative dogs.

Visceral leishmaniasis in an environmentally protected area in southeastern Brazil: Epidemiological and laboratory cross-sectional investigation of phlebotomine fauna, wild hosts and canine cases.

Background Leishmaniasis is a rapidly expanding zoonosis that shows increasing urbanization. Concern exists regarding the role of wildlife in visceral leishmaniasis (VL) transmission, due to frequent natural or anthropogenic environmental changes that facilitate contact between wildlife, humans and their pets. The municipality of Campinas, in southeastern Brazil, initially recorded VL in 2009, when the first autochthonous case was confirmed in a dog living in an upscale residential condominium, located inside an environmentally protected area (EPA). Since then, disease transmission remains restricted to dogs inhabiting two geographically contiguous condominiums within the EPA. Methodology/Principal findings We conducted a cross-sectional study of the VL focus to investigate Leishmania spp. infection in domestic dogs, wild mammals and sand flies using molecular tools and recommended serological techniques. Canine seroprevalences of 1.5% and 1.2% were observed in 2013 and 2015, respectively. Six insect species, confirmed or suspected vectors or potential transmitters of Leishmania, were identified. Two specimens of the main L. (L.) infantum vector in Brazil, Lutzomyia longipalpis, were captured in the EPA. Natural infection by L. (L.) infantum was recorded in one Expapillata firmatoi specimen and two Pintomyia monticola. Natural infection by L. (L.) infantum and Leishmania subgenus Viannia was also detected in two white-eared opossums (Didelphis albiventris), a known reservoir of VL. Geographical coordinates of each sampling of infected animals were plotted on a map of the EPA, demonstrating proximity between these animals, human residences, including the dogs positive for VL, and forest areas. Conclusions/Significance The EPA, which is inhabited by humans, has an active VL focus. The risk of establishing and maintaining disease transmission foci in similar scenarios, i.e. wild areas that undergo environmental modifications, is evident. Moreover, different epidemiological profiles of VL must be included to elaborate prevention and control measures that consider the particularities of each transmission area.

Antibodies and Molecular Detection of Leishmania (Leishmania) infantum in Samples of Free-Ranging Marmosets (Primates: Callitrichidae: Callithrix spp.) in an Area of Canine Visceral Leishmaniasis in Southeastern Brazil

Leishmaniasis is a vector-borne parasitic protozoan infection that affects mammals and involves a complex epidemiology. Although dogs are considered the main reservoir in zoonotic visceral leishmaniasis (VL), the possible presence of other mammalian species acting as reservoirs has been associated as a possible cause of lack of success in the control of human VL in many endemic areas. The knowledge about natural infections of some species is still scarce, such as nonhuman primates (NHP), especially from the genus Callithrix (marmosets). We investigated the infection by Leishmania (Leishmania) infantum, the agent of VL in the Americas, in 26 marmosets captured monthly, from April 2014 to March 2015, in an environmentally protected area (EPA) in Southeastern Brazil. The EPA has undergone significant environmental changes and has a transmission focus of canine VL since 2009. Serology was performed through the direct agglutination test, which detected low antibody titers in seven marmosets (7/26; 26.9%, 95% confidence interval 9.9-44.0), being five Callithrix penicillata (black-tufted-ear marmoset) and two Callithrix jacchus (white-tufted-ear marmoset). The presence of the DNA of Leishmania was investigated in blood and skin samples by PCR and genetic sequencing. This is the first report of the detection of L. (L.) infantum in the skin of a marmoset, which was verified in a sample from one C. penicillata. The results demonstrate the natural infection of marmosets by L. (L.) infantum and may suggest the participation of these animals as hosts in the parasites transmission cycle in the EPA. However, more comprehensive studies are needed to elucidate their role on the VL epidemiology in this area and also in different endemic areas, especially because these NHP are increasingly in contact with humans and domestic animals, particularly due to environmental changes.

O xenodiagnóstico na doença de Chagas: influência do sexo dos triatomíneos

Foi avaliado, comparativamente, o desempenho de femeas e machos de Triatoma infestans, em 5o estadio de desenvolvimento, em xenodiagnosticospara a tripanossomiase americana. Estes exames foram realizados em 40 pacientes, em fase cronica da doenca de Chagas. Em cada exame utilizou-se 20 ninfas femeas e 20 ninfas machos, que foram pesadas imediatamente antes e apos o repasto. O exame individual do conteudo intestinal dos triatomineos foi efetuado aos 20, 30, 60 e 90 dias apos a aplicacao dos xenodiagnosticos. As ninfas femeas e machos, ingeriram, em media, 230 e210 mg de sangue, respectivamente. Os resultados revelaram 15 (37,5%) xenodiagnosticos positivos; todavia, em nenhum destes exames foi verificada positivacao, para T. cruzi, de todas as ninfas utilizadas. As ninfas femeas ingeriram significativamente mais sangue que as ninfas machos, isto vem de encontro com as observacoes de maior positividade, para T. cruzi entre as femeas, sugerindo, serem estas ninfas capazes de possibilitar maior incremento a sensibilidade do xenodiagnostico na fase cronica da doenca de Chagas, sendo necessarios novos estudos para confirmacao desta hipotese.