In this study, we identified the phlebotomine sandfly vectors involved in the transmission of American Cutaneous Leishmaniasis (ACL) in Assis Brasil, Acre, Brazil, which is located on the Brazil-Peru-Bolivia frontier. The genotyping of Leishmania in phlebotomines was performed using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism. A total of 6,850 sandflies comprising 67 species were captured by using CDC light traps in rural areas of the municipality. Three sandfly species were found in the state of Acre for the first time: Lutzomyia georgii, Lu. complexa and Lu. evangelistai. The predominant species was Lu. auraensis/Lu. ruifreitasi and Lu. davisi (total 59.27%). 32 of 368 pools were positive for the presence of Leishmania DNA (16 pools corresponding to Lu. davisi, and 16 corresponding to Lu. auraensis/Lu. ruifreitasi), with a minimal infection prevalence of 1.85% in Lu. davisi and 2.05% in Lu. auraensis/Lu. ruifreitasi. The Leishmania species found showed maximum identity with L. (Viannia) guyanensis and L. (V.) braziliensis in both phlebotomine species. Based on these results and similar scenarios previously described along the Brazil/Peru/Bolivia tri-border, the studied area must take into consideration the possibility of Lu. davisi and Lu. auraensis/Lu. ruifreitasi as probable vectors of ACL in this municipality.

BackgroundPhlebotomine sand flies (Diptera: Psychodidae) are insects of medical importance due to the role that some species play in the transmission of leishmaniasis. This work aimed to study some ecological aspects among sand flies fauna inhabiting two different environments: the várzea (lowland Amazonian forest) and terra firme (upland Amazonian forest), both located in Tefé Municipality, Amazonas State, Braziland to detect Leishmania infection in those phlebotomine populations.MethodsSand flies were collected using HP light traps. Collection took place over the course of six months: January, February, April, August, September, and October of 2013. To detect natural infection by Leishmania, DNA samples were extracted from female sand flies and submitted to Polymerase Chain Reaction (PCR) targeting the kDNA gene; Leishmania species were identified by PCR-RFLP targeting the hsp70 gene and genetic sequencing.ResultsIn all, 5,716 individuals were collected, and 46 species were identified. Trichophoromyia ubiquitalis (3,330 – 58.26%) and Nyssomyia antunesi (661 – 11.26%) were the most abundant species. Species richness was greater in terra firme environments (42 species) than in the várzea environments (22 species), and forests ecotopes (43 species) were richer than peridomiciles (28 species). DNA of Leishmania was found in Th. ubiquitalis and Psychodopygus davisi, both of which inhabit the terra firme environment and sequencing analysis confirmed the presence of Leishmania (Viannia) lainsoni DNA in Th. ubiquitalis in Tefé Municipality.ConclusionsThe high abundance of Th. ubiquitalis and Ps. davisi and detection of DNA of Leishmania sp. may indicate that both species could be putative vectors for American Cutaneous Leishmaniasis (ACL) in the terra firme environment of Tefé. The sand fly fauna found in várzea is rich and diverse, exhibiting several species, nevertheless the seasonal hydric stress during part of the year that could influence the local diversity, if compared with other studies. This is the first report in Amazonas State of Th. ubiquitalis with presence of L. (V.) lainsoni DNA.

SUMMARYIn this study, Leishmaniaspecies were identified by Polymerase Chain Reaction (PCR). The epidemiology of patients suspected of having American Cutaneous Leishmaniasis in the municipality of Assis Brasil, Acre State, located in the Brazil/Peru/Bolivia triborder was also investigated. By PCR, the DNA of Leishmaniawas detected in 100% of the cases (37 samples) and a PCR-Restriction Fragment Length Polymorphism (RFLP) of the hsp 70gene identified the species in 32 samples: Leishmania (Viannia) braziliensis (65.6%) , L. (V.) shawi (28.1%) , L. (V.) guyanensis (3.1%) and mixed infection L. (V.) guyanensis and L. (Leishmania) amazonensis (3.1%)This is the first report of L. (V.) shawiand L. (L.) amazonensis in Acre. The two predominant species were found in patients living in urban and rural areas. Most cases were found in males living in rural areas for at least three years and involved in rural work. This suggests, in most cases, a possible transmission of the disease from a rural/forest source, although some patients had not engaged in activities associated with permanence in forestall areas, which indicate a possible sandflies adaptation to the periurban setting.

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This study evaluated the species composition of sand flies and identified potential vectors of Leishmania Ross species in rural areas of Porto Velho, Rondônia State, Brazil. American cutaneous leishmaniasis (ACL) is one of the gravest threats to public health in this state. Sand flies were collected over the course of 2014 and 2015 using HP light traps. Polymerase chain reaction was performed by targeting the Leishmania mkDNA region. In total, 2,344 sand flies were collected, from which 45 species, nine subgenera, and five species group were identified. The most abundant species were Lutzomyia antunesi (Coutinho) (n = 597, 25.47%), Lutzomyia ubiquitalis (Mangabeira) (n = 496, 21.16%), and Lutzomyia octavioi (Vargas) (n = 199, 8.49%). The greatest diversity occurred in the forest environment where the most abundant species were L. antunesi (n = 588, 25.07%), L. ubiquitalis (n = 493, 21.02%), L. octavioi (n = 199, 8.49%), and Lutzomyia flaviscutellata (Mangabeira) (n = 132, 5.63%). Two pools of L. ubiquitalis were positive for Leishmania DNA, which suggests that L. ubiquitalis is a putative vector of leishmaniasis in the municipality of Porto Velho.

O presente trabalho descreve a atividade do extrato etanólico (EE) dos frutos de Combretum leprosum, do triterpeno 3b, 6b, 16b-triidroxilup-20(29)-eno (1) e seus derivados sintéticos (1a-1d), sobre promastigotas de Leishmania amazonensis. O EE apresentou atividade leishmanicida e o valor de IC 50 foi de 24,8 mg mL -1 . Já o triterpeno 3b, 6b, 16b-trihidroxilup-20(29)-eno (1), na concentração de 5,0 mg mL -1 , apresentou uma potente ação inibitória sobre a proliferação das promastigotas (IC 50 = 3,3 mg mL -1 ). Entre os derivados sintéticos, apenas 1b e 1d apresentaram atividade contra as promastigotas (IC 50 = 3,48 mg mL -1 e 5,8 mg mL -1 , respectivamente). Por outro lado, o derivado sintético 1a não apresentou atividade sobre as promastigotas de L. amazonensis. O EE, (1) e os derivados sintéticos 1a-1d não apresentaram efeito citotóxico sobre macrófagos peritoneais de camundongos. Estes resultados fornecem evidencias de que o extrato etanólico e o lupano isolado de C. leprosum possui atividade contra promastigotas de L. amazonensis, podendo ser utilizados como ferramentas no estudo de novas drogas leishmanicidas. This paper describes the activity of the ethanolic extract (EE), obtained from the fruits of Combretum leprosum, the triterpene 3b, 6b, 16b-trihydroxylup-20(29)-ene (1) and its synthetic derivatives 1a-1d on Leishmania amazonensis promastigotes. The EE displayed leishmanicidal activity and the IC 50 was 24.8 mg mL -1 . However, the triterpene 3b, 6b, 16b-trihydroxylup-20(29)-ene (1), at a concentration of 5.0 mg mL -1 , showed a potent inhibitory activity on promastigotes proliferation (IC 50 = 3.3 mg mL -1 ). Among the synthetic derivatives, only (1b) and (1d) were active against promastigotes (IC 50 = 3.48 mg mL -1 and 5.8 mg mL -1 , respectively). Moreover, the synthetic derivative 1a showed no activity on promastigotes of L. amazonensis. EE, (1) and the synthetic derivatives 1a-1d showed no cytotoxic effect on mice peritoneal macrophages. These results provide evidence that the ethanolic extract and the lupane isolated from C. leprosum was active against promastigotes of L. amazonensis, and may be used as a tool in the studies of new antileishmanial drugs. The clinical manifestations of leishmaniasis are often divided in cutaneous, diffuse cutaneous, mucocutaneous and visceral leishmaniasis. Keywords3,4 Cutaneous leishmaniasis can Teles et al. 937 Vol. 22, No. 5, 2011 be spontaneously healed after a few months, or, depending on the Leishmania species, develop into diffuse cutaneous, relapsing cutaneous or mucocutaneous leishmaniasis. Visceral leishmaniasis, if untreated, leads to death in most patients. 5,6 This disease causes considerable morbidity and severe face-disfigurement lesions on the affected people.Nowadays, chemotherapy for leishmaniasis is still based on pentavalent antimonials (Glucantime and Pentostam), diamines (Pentamidine) and antifungal polyene (Amphotericin B). These are only a few of the drugs available since 1940. Unfortunately, they are generally toxic, expensive, s...

The Micrurus genus is the American representative of Elapidae family. Micrurus spixii is endemic of South America and northern states of Brazil. Elapidic venoms contain neurotoxins that promote curare-mimetic neuromuscular blockage. In this study, biochemical and functional characterizations of M. spixii crude venom were performed and a new neurotoxic phospholipase A2 called MsPLA2-I was isolated. M. spixii crude venom caused severe swelling in the legs of tested mice and significant release of creatine kinase (CK) showing its myotoxic activity. Leishmanicidal activity against Leishmania amazonensis (IC50 1.24 μg/mL) was also observed, along with antiplasmodial activity against Plasmodium falciparum, which are unprecedented for Micrurus venoms. MsPLA2-I with a Mr 12,809.4 Da was isolated from the crude venom of M. spixii. The N-terminal sequencing of a fragment of 60 amino acids showed 80% similarity with another PLA2 from Micrurus altirostris. This toxin and the crude venom showed phospholipase activity. In a mouse phrenic nerve-diaphragm preparation, M. spixii venom and MsPLA2-I induced the blockage of both direct and indirect twitches. While the venom presented a pronounced myotoxic activity, MsPLA2-I expressed a summation of neurotoxic activity. The results of this study make M. spixii crude venom promising compounds in the exploration of molecules with microbicidal potential.

RESUMOAs formigas são consideradas insetos vetores de várias espécies de patógenos e apresentam grande adaptabilidade a diversos tipos de ambientes. Por essa razão podem oferecer risco a pacientes imunodeprimidos, em ambiente hospitalar. O objetivo deste estudo foi verificar o perfil das bactérias Gram positivas veiculadas por formicídeos em ambiente hospitalar. Os insetos foram coletados na sala de emergência, no setor de pneumologia e no de gastroenterologia em um hospital público de Porto Velho, Estado de Rondônia, Brasil, no período de março a junho de 2012. As bactérias isoladas foram caracterizadas quanto à coloração de Gram, aos testes para identificação de bactérias do gênero Staphylococcus, Streptococcus e Micrococcus e quanto ao perfil de suscetibilidade a antimicrobianos. Entre as 130 formigas capturadas, 62% das cepas extraídas das formigas foram identificadas como pertencentes ao gênero Staphylococcus, 25% Streptococcus e 13% Micrococcus. Em relação à resistência aos antibióticos, 83% das cepas foram resistentes à oxacilina e 50% à vancomicina. Conclui-se que as formigas apresentaram grande potencial como veiculadoras mecânicas de bactérias, sugerindo o risco de infecção nosocomial e a permanência de cepas com alta resistência no ambiente hospitalar. 33 INTRODUÇÃOO processo de urbanização promoveu o aumento de doenças causadas e/ou transmitidas por artrópodes, entre esses, as formigas, um grupo que obteve melhor adaptação ao ambiente urbano, devido à grande disponibilidade de abrigo e alimento para si, o que facilita seu estabelecimento no ambiente e sua proliferação 1 . É importante ressaltar que, atualmente, cerca de 12.500 espécies de formigas já foram descritas, mas estima-se a existência de cerca de 21 mil espécies, e, dessas, menos de 20 espécies podem ser classificadas como pragas urbanas no Brasil 2 .As formigas realizam simbiose com bactérias, sendo consideradas vetores mecânicos de várias espécies desses microrganismos 3,4 . Além disso, possuem uma grande capacidade de locomoção, chegando a percorrer 3 cm por segundo, e, por isso, podem transitar em vários locais, como UTI, leitos, cozinhas, salas cirúrgicas e outros ambientes hospitalares, muitas vezes sem serem notadas. Desta forma, representam um fator de risco à saúde pública, uma vez que aumentam o risco de infecções hospitalares 5 .Sua presença nesse tipo de local pode ocorrer devido à estrutura arquitetônica dos hospitais, a sua localização próxima a residências, às embalagens de alguns medicamentos que podem trazer ninhos de formigas e ao grande fluxo de pessoas e alimentos que servem como atrativo para elas 6 .Bacterias Gram-positivas vehiculadas por hormigas en ambiente hospitalario en la Ciudad de Porto Velho, Estado de Rondônia, BrasilGram-positive bacteria carried by ants in hospital environment in the City of Porto