Ana Maria Bonetti

Esterase enzymes involved in pyrethroid and organophosphate resistance in a Brazilian population of Riphicephallus (Boophilus) microplus (Acari, Ixodidae).

Esterases are a group of enzymes that are reportedly associated with acaricide resistance in Riphicephallus (Boophilus) microplus. A comparative analysis was made of the esterase patterns in malathion and deltamethrin-sensitive, tolerant and resistant tick groups, using non-denaturing polyacrylamide gel electrophoresis. Electrophoretical profiles revealed four bands of esterase activity against alpha-naphthyl acetate; which were dubbed EST-1 to EST-4. The EST-3 and EST-4 were detected in all strains and were classified as carboxylesterases (CaEs). The EST-2, classified as an acetylcholinesterase (AChE), was detected in all groups, but its staining intensity increased from susceptible to resistant groups, indicating an altered production according to the degree of resistance. EST-1, which was also classified as an AChE, was detected exclusively in tolerant and resistant groups to both acaricides, but displayed greater activity in the malathion-resistant group. These data suggest that these AChEs may represent an important detoxification strategy developed to overcome the effects of acaricides.

Urban Ants and Transportation of Nosocomial Bacteria

Many ant species displaying synanthropic behavior that have successfully dispersed in urban areas can cause problems in hospitals by acting as bacterial vectors. In this study, we encountered bacteria on ants collected at the Universidade Federal de Uberlândia hospital, in the campus and at households nearby. The ants were identified as Tapinoma melanocephalum (Fabricius) and Camponotus vittatus (Forel) (Hymenoptera: Formicidae) and the bacterial strains found here belong to the group of the coagulase-positive staphylococcus, coagulase-negative staphylococcus and gram negative bacilli, including antimicrobial drug-resistant strains. An investigation of the bacteria found in the ants and in the environment revealed that some ants carried non-isolated bacteria from the same environment and with high levels of resistance, evidencing the transmission potential of these insects.

Esterase profile in a pyrethroid-resistant Brazilian strain of the cattle tick Boophilus microplus (Acari, Ixodidae)

The cattle tick Boophilus microplus causes great damage in livestock and is considered one of the most important tropical ectoparasites. The traditional method of control is based on the intensive use of pesticides, however the indiscriminate use of these compounds over the years has led to the selection of resistant ticks. Hydrolases, particularly esterases (EST), have been reported to be associated with acaricide resistance in B. microplus. We compared the esterase profile of susceptible and cypermethrin-resistant strains of adult B. microplus and a pyrethroid susceptible reference strain (the Mozzo strain) using polyacrylamide gel electrophoresis and specific staining. The electrophoretic profiles of protein extracts revealed the presence of four regions with esterase activity in the cypermethrin-resistant strain and three of these regions in the susceptible strains. The bands were numbered EST-1 to EST-4 in sequence (starting from the anode) according to their decrease in negative charge. The EST-1A and EST-1B enzymes were detected only in the resistant strain. The inhibition studies with eserine sulfate, copper sulfate, p- p-chloromercuribenzoate (pCMB), malathion and phenylmethylsulfonyl fluoride (PMSF) indicated that the EST-1A and EST-1B enzymes belong to the acetylcholinesterase class and are probably associated with resistance to acaricides in this Brazilian resistant strain of B. microplus.

Circulating hemocytes from larvae of Melipona scutellaris (Hymenoptera, Apidae, Meliponini): cell types and their role in phagocytosis.

Infection in insects stimulates a complex defensive response. Recognition of pathogens may be accomplished by plasma or hemocyte proteins that bind specifically to bacterial or fungal polysaccharides. Several morphologically distinct hemocyte cell types cooperate in the immune response. Hemocytes attach to invading organisms and then isolate them by phagocytosis, by trapping them in hemocyte aggregates called nodules, or by forming an organized multicellular capsule around large parasites. In the current investigation the cellular in the hemolymph third instar larvae of M. scutellaris has been characterized by means of light microscopy analysis and phagocytosis assays were performed in vivo by injection of 0.5 microm fluorescence beads in order to identify the hemocyte types involved in phagocytosis. Four morphotypes of circulating hemocytes were found in 3rd instar larvae: prohemocytes, plasmatocytes, granulocytes and oenocytoids. The results presented plasmatocytes and granulocytes involved in phagocytic response of foreign particles in 3rd instar larvae of M. scutellaris.

Levantamento da diversidade de formigas (Hymenoptera: Formicidae) na região urbana de Uberlândia, MG

Uberlândia, MG, Brazil, underwent an accelerated process of urbanization with a population growth of 3,54% each year, higher than the national average. One of the problems emergent from urbanization is the use of different habitats for a great variety of insects. The objective of this study was to identify species of house-invading ants that occur in the urban area of Uberlândia. The occurrence of the house-invading species in regard to the time of urbanization, neighborhood infrastructure, age and maintenance of private houses was also assessed. The ants were collected using bait-traps in 120 residences from 12 neighborhoods which were put in three groups. Fourteen species of ants were catalogued, with Camponotus (Mayr), Monomorium (Mayr) and Tapinoma (Foerster) being the most frequent genera. Only Camponotus vittatus (Forel), Monomorium pharaonis (L.), Tapinoma melanocephalum (Fabricius) and Brachymyrmex sp. (Mayr) were collected in all three groups. The relation between the ants collected and the age and maintenance of the private houses showed that Paratrechina longicornis (Latreille) was most frequent (60%) in poorly preserved or precarious constructions. Whereas, C. vittatus and Camonotus melanoticus (Emery) occurred in all categories of maintenance. Overall, C. vittatus which had not been found in any previously published survey of urban ants, was the most frequent species in urban areas of the Cerrado.

Analysis of the genetic diversity of vancomycin-resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) are among the most frequent causes of hospital infections worldwide, thus justifying the increasing use of vancomycin. In this study, we evaluated the presence of glycopeptide-resistant staphylococci, in 41 patients hospitalized in the Clinical Hospital of the Federal University of Uberlândia in Uberlândia, MG, who were being treated with vancomycin. All isolates were plated on Mueller-Hinton agar containing vancomycin. Vancomycin resistance was confirmed by surface growth after incubation for 24-48 h at 35oC. Heteroresistance was evaluated by plating with a large inoculum (108 CFU/mL). One patient with nephritis who was on a hemodialysis program was diagnosed with the phenotype isolate of vancomycin-intermediate Staphylococcus aureus (VISA) (CIM = 8 mg/mL) and in eight patients, strains of heteroresistant Staphylococcus corresponding to the hVISA phenotype were isolated. In addition to the extended use of vancomycin, other risk factors associated with the presence of these microorganisms included the use of three or more antimicrobial agents, surgery, and three or more invasive procedures. Molecular analysis by randon amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) showed two clusters involving two samples each one of them, in surgical patients, with temporal and spatial relationship and isolates similarity concerning the susceptibility range to antimicrobial agents.

Pollen and stomata morphometrics and polyploidy in Eriotheca (Malvaceae-Bombacoideae)

Approximately 70% of the angiosperm species are polyploid, an important phenomenon in the evolution of those plants. But ploidy estimates have often been hindered because of the small size and large number of chromosomes in many tropical groups. Since polyploidy affects cell size, morphometric analyses of pollen grains and stomata have been used to infer ploidy level. Polyploidy is present in many species of the Cerrado, the Neotropical savanna region in Central Brazil, and has been linked to apomixis in some taxa. Eriotheca gracilipes and Eriotheca pubescens are common tree species in this region, and present cytotypes that form reproductive mosaics. Hexaploid individuals (2n = 6x = 276) are polyembryonic and apomictic, while tetraploid and diploid individuals (2n = 2x = 92, 2n = 4x = 184) are sexual and monoembryonic. We tested whether morphometric analysis can be used to estimate ploidy levels in E. gracilipes and E. pubescens individuals. Pollen material from diploid and hexaploid individuals of E. gracilipes, and tetraploid and hexaploid individuals of E. pubescens, were fixed in 50% FAA, and expanded leaves were dried in silica gel. Pollen grains and stomata of at least five individuals from each population were measured. The results demonstrate that all measures were significantly different among cytotypes. Individuals with higher levels of ploidy (hexaploid) all presented measurements that were higher than those with lower levels (diploid and tetraploid). There was no overlap between ploidy levels in each species at 95% confidence interval. Thus, the size of the pollen grains and stomata are effective parameters for analysis of ploidy levels in E. gracilipes and E. pubescens.

Insights into the Melipona scutellaris (Hymenoptera, Apidae, Meliponini) fat body transcriptome

The insect fat body is a multifunctional organ analogous to the vertebrate liver. The fat body is involved in the metabolism of juvenile hormone, regulation of environmental stress, production of immunity regulator-like proteins in cells and protein storage. However, very little is known about the molecular mechanisms involved in fat body physiology in stingless bees. In this study, we analyzed the transcriptome of the fat body from the stingless bee Melipona scutellaris. In silico analysis of a set of cDNA library sequences yielded 1728 expressed sequence tags (ESTs) and 997 high-quality sequences that were assembled into 29 contigs and 117 singlets. The BLAST X tool showed that 86% of the ESTs shared similarity with Apis mellifera (honeybee) genes. The M. scutellaris fat body ESTs encoded proteins with roles in numerous physiological processes, including anti-oxidation, phosphorylation, metabolism, detoxification, transmembrane transport, intracellular transport, cell proliferation, protein hydrolysis and protein synthesis. This is the first report to describe a transcriptomic analysis of specific organs of M. scutellaris. Our findings provide new insights into the physiological role of the fat body in stingless bees.

Esterase profile in the postembryonic development of Rhipicephalusmicroplus

The objective of this work was to analyze the pattern of esterase activity in the development stages of Rhipicephalus microplus by nondenaturing polyacrylamide gel electrophoresis using specific staining for esterase. The electrophoretical results revealed the presence of nine regions displaying esterase activity, stained with both alpha-naphthyl acetate and beta-naphthyl acetate, and classified as alpha-beta-esterase. Stage-specific esterases were found, with the first nymphal and larval stages showing the greatest esterase activity throughout the development. An esterase called EST-4 was detected only in males and was considered sex-specific. There are differences in the esterase profile among the different postembryonic development stages of R. microplus.

Sex determination in bees. XXX. Effects of juvenile hormone on the development of tergal glands in Melipona

SUMMARYWorker larvae of Melipona rufiventris, M. quadrifasciata and M. compressipes were treated topically with juvenile hormone (JH) during the cocoon-spinning phase. Examination of the tergites of the queens obtained following JH application showed induced differentiation into adults with full female (queen) traits. Melipona workers had tergal glands only in tergite II, whereas natural and JH-induced queens had tergal glands in tergite II plus additional glands in at least one other of tergites III to VII.