Alexandre de Almeida e Silva

Antimicrobial peptides from Phyllomedusa frogs: from biomolecular diversity to potential nanotechnologic medical applications

Screening for new bioactive peptides in South American anurans has been pioneered in frogs of the genus Phyllomedusa. All frogs of this genus have venomous skin secretions, i.e., a complex mixture of bioactive peptides against potential predators and pathogens that presumably evolved in a scenario of predator–prey interaction and defense against microbial invasion. For every new anuran species studied new peptides are found, with homologies to hormones, neurotransmitters, antimicrobials, and several other peptides with unknown biological activity. From Vittorio Erspamer findings, this genus has been reported as a “treasure store” of bioactive peptides, and several groups focus their research on these species. From 1966 to 2009, more than 200 peptide sequences from different Phyllomedusa species were deposited in UniProt and other databases. During the last decade, the emergence of high-throughput molecular technologies involving de novo peptide sequencing via tandem mass spectrometry, cDNA cloning, pharmacological screening, and surface plasmon resonance applied to peptide discovery, led to fast structural data acquisition and the generation of peptide molecular libraries. Research groups on bioactive peptides in Brazil using these new technologies, accounted for the exponential increase of new molecules described in the last decade, much higher than in any previous decades. Recently, these secretions were also reported as a rich source of multiple antimicrobial peptides effective against multidrug resistant strains of bacteria, fungi, protozoa, and virus, providing instructive lessons for the development of new and more efficient nanotechnological-based therapies for infectious diseases treatment. Therefore, novel drugs arising from the identification and analysis of bioactive peptides from South American anuran biodiversity have a promising future role on nanobiotechnology.

The Dynamics of Transmission and Spatial Distribution of Malaria in Riverside Areas of Porto Velho, Rondônia, in the Amazon Region of Brazil

The study area in Rondônia was the site of extensive malaria epidemic outbreaks in the 19th and 20th centuries related to environmental impacts, with large immigration flows. The present work analyzes the transmission dynamics of malaria in these areas to propose measures for avoiding epidemic outbreaks due to the construction of two Hydroelectric Power Plants. A population based baseline demographic census and a malaria prevalence follow up were performed in two river side localities in the suburbs of Porto Velho city and in its rural vicinity. The quantification and nature of malaria parasites in clinical patients and asymptomatic parasite carriers were performed using microscopic and Real Time PCR methodologies. Anopheles densities and their seasonal variation were done by monthly captures for defining HBR (hourly biting rate) values. Main results: (i) malaria among residents show the riverside profile, with population at risk represented by children and young adults; (ii) asymptomatic vivax and falciparum malaria parasite carriers correspond to around 15% of adults living in the area; (iii) vivax malaria relapses were responsible for 30% of clinical cases; (iv) malaria risk for the residents was evaluated as 20–25% for vivax and 5–7% for falciparum malaria; (v) anopheline densities shown outdoors HBR values 5 to 10 fold higher than indoors and reach 10.000 bites/person/year; (vi) very high incidence observed in one of the surveyed localities was explained by a micro epidemic outbreak affecting visitors and temporary residents. Temporary residents living in tents or shacks are accessible to outdoors transmission. Seasonal fishermen were the main group at risk in the study and were responsible for a 2.6 fold increase in the malaria incidence in the locality. This situation illustrates the danger of extensive epidemic outbreaks when thousands of workers and secondary immigrant population will arrive attracted by opportunities opened by the Hydroelectric Power Plants constructions.

Amazonian biodiversity: a view of drug development for Leishmaniasis and malaria

Chemotherapy is the only validated therapy for the treatment for the neglected diseases leishmaniasis and malaria. However, the emergence of drug resistance, collateral effects and long-term treatment encourage the development of new and more efficient drugs. The Amazon tropical forest includes the richest areas of biodiversity in the world, including a great number of microbes, plant and animal species that produce a source of interesting biologically active molecules. Several of these molecules, obtained from plant extracts and frog venom have leishmanicidal and plasmodicidal activity, highlighting the potential of this biodiversity for the development of new drugs. In research, modern approaches in new drug development are carried out using combinatorial chemistry, high-throughput screening, bioinformatics, molecular interaction, crystallography and dynamic studies of cellular and systemic toxicity. In Brazil, these techniques are mainly present in only a few academic groups with no efficient connection to industry. The problem associated with over-regulation for accessing the biological material in restricted areas, local populations and indigenous areas places major barriers in the path of research and development of new drugs. Thus, the association of academic research groups in Brazil, encouraged and supported by government and industry, is essential to overcome these major barriers related to the development of new products for treatment of neglected diseases from Amazonian biodiversity in future years.

Malaria and hematological aspects among residents to be impacted by reservoirs for the Santo Antônio and Jirau Hydroelectric Power Stations, Rondônia State, Brazil

In Rondônia State, Brazil, two new hydroelectric plants, Santo Antônio and Jirau, are scheduled for construction on the Madeira River, upriver from the State capital, Porto Velho. The current study analyzes malaria prevalence before the construction and provides information on the possible impacts of malaria burden related to the influx of thousands of persons attracted by direct and indirect employment opportunities. According to the findings, malaria is present throughout the region, with varying prevalence rates. The existence of potential asymptomatic malaria carriers among the local population may be epidemiologically relevant and should be considered in the malaria control programs organized by public authorities and companies responsible for building the power plants, aimed at early diagnosis and treatment, vector control, water supply, and infrastructure in the urban areas.

Mosquito abundance and behavior in the influence area of the hydroelectric complex on the Madeira River, Western Amazon, Brazil

Malaria is currently highly prevalent and restricted to the north of Brazil, and its dynamics are severely affected by human environmental changes, such as the large dam construction recently approved by the Brazilian Government in Rondônia. We studied the mosquito fauna and behavior before hydroelectric construction. Mosquitoes were captured by human landing catches on the riversides of the Madeira River in Porto Velho, Rondônia. A total of 3121 mosquitoes from eight different genera were collected; only Mansonia and Anopheles darlingi were found in all 21 collection sites throughout the night. These results suggest that the riverines of the study area are exposed to malaria.

Species structure of sand fly (Diptera: Psychodidae) fauna in the Brazilian western Amazon

We surveyed areas of the state of Rondônia in western Amazon for phlebotomine, which are potential vectors of leishmaniasis. A total of 5,998 specimens were captured, resulting in the identification of 48 species within the Lutzomyia (99.98%) and Brumptomyia (0.02%) genera. The predominant species was Lutzomyia davisi, followed by Lutzomyia umbratilis, Lutzomyia llanosmartinsi, Lutzomyia c. carrerai, Lutzomyia dendrophyla, Lutzomyia nevesi and Lutzomyia whitmani. All sand flies identified as vectors for cutaneous leishmaniasis in Brazil, i.e., Lu. davisi, Lu. umbratilis, Lu. c. carrerai and Lu. whitmani, were found in the surveyed areas.

Larvicidal activity of natural and modified triterpenoids against Aedes aegypti (Diptera: Culicidae).

BACKGROUND Insecticide resistance to commonly used substances demands new molecules for the chemical control of the dengue vector Aedes aegypti. Because natural product sources have been an alternative to obtain larvicidal compounds, the aim of this study was to evaluate the triterpenoids betulinic (BA) and ursolic (UA) acids and their semi-synthetic derivatives against larval Ae. aegypti. BA, UA, ten derivatives modified at the C-3 position and a positive control (diflubenzuron) were evaluated. Larvicidal assays were carried out with early fourth-instar larvae, and mortality was observed between 48 and 96 h. Doses from 200 to 10 ppm were used to calculate lethal concentrations (LCs). RESULTS Natural compounds, i.e. UA and BA, had the lowest LCs (LC50 of 112 and 142 ppm respectively), except for the modified compound 2b (LC50 of 130 ppm). Larvicidal activity increased significantly from 48 to 96 h for all the compounds evaluated, ranging from 20 to 50% after 48 h and from 48 to 76% after 96 h. Some derivatives, e.g. 2a and 2d, had up to a three-fold larvicidal activity increase from 48 to 96 h. CONCLUSION BA, UA and their derivatives showed larvicidal activity against Ae. aegypti larvae, increasing significantly from 48 to 96 h. The presence of a hydroxyl group is essential for larvicidal potential in these triterpenoids.

Cerambycidae (Insecta: Coleoptera) of the Parque Natural Municipal de Porto Velho, Rondônia, Western Amazon, Brazil

Cerambycidae is one of the largest families of beetles and it is estimated that there are about 25,000 species of longhorned beetles in the world. However, little is known about the distribution of many species in some regions, including the Amazon. The longhorned beetles are indispensable in the ecological chain, contributing mostly to the recycling of dead wood in forest. The present study is an inventory of the Cerambycidae of the Parque Natural Municipal de Porto Velho, Rondonia, Brazil. Insects were sampled from June 2008 to May 2009 using Malaise and light traps. A total of 61 species were identified, of which 33 are new records for Rondonia and one for Amazon region, i.e., Anisopodus melzeri Gilmour, 1965.

Change in Anopheles richness and composition in response to artificial flooding during the creation of the Jirau hydroelectric dam in Porto Velho, Brazil

BackgroundAnopheles mosquitoes are the only vectors of human malaria. Anopheles species use standing water as breeding sites. Human activities, like the creation of an artificial lake during the implementation of hydroelectric power plants, lead to changes in environmental characteristics and, therefore, may changes the species richness and composition of Anopheles mosquitoes. The aim of the present study was to verify whether or not there is an association between the artificial flooding resulting from the construction of the Jirau hydroelectric power plant, and the richness and composition of anophelines.MethodsMosquitoes samples were obtained monthly from the Jirau hydroelectric power plant area located at Porto Velho, Rondônia State, using Human Landing Catch (06:00–10:00 PM). Mosquitoes collected were transported to Laboratório de Entomologia Médica FIOCRUZ-RO where they were identified until species using dichotomous key.ResultsA total of 6347 anophelines belonging to eight different species were collected. The anophelines species richness was significantly lower during the first flooding stage. Differences in anophelines species composition were found when comparing the first flooding stage with the other stages. Furthermore, the mean number of Anopheles darlingi, the main vector of malaria in the region, increases during the first and the third flooding stages.ConclusionsThe continual monitoring of these vectors during the late operational phase may be useful in order to understand how anophelines will behave in this area.

Biochemical and Functional Characterization of Parawixia bistriata Spider Venom with Potential Proteolytic and Larvicidal Activities

Toxins purified from the venom of spiders have high potential to be studied pharmacologically and biochemically. These biomolecules may have biotechnological and therapeutic applications. This study aimed to evaluate the protein content of Parawixia bistriata venom and functionally characterize its proteins that have potential for biotechnological applications. The crude venom showed no phospholipase, hemorrhagic, or anti-Leishmania activities attesting to low genotoxicity and discrete antifungal activity for C. albicans. However the following activities were observed: anticoagulation, edema, myotoxicity and proteolysis on casein, azo-collagen, and fibrinogen. The chromatographic and electrophoretic profiles of the proteins revealed a predominance of acidic, neutral, and polar proteins, highlighting the presence of proteins with high molecular masses. Five fractions were collected using cation exchange chromatography, with the P4 fraction standing out as that of the highest purity. All fractions showed proteolytic activity. The crude venom and fractions P1, P2, and P3 showed larvicidal effects on A. aegypti. Fraction P4 showed the presence of a possible metalloprotease (60 kDa) that has high proteolytic activity on azo-collagen and was inhibited by EDTA. The results presented in this study demonstrate the presence of proteins in the venom of P. bistriata with potential for biotechnological applications.