Abstract Background: Among the hymenopteran insect venoms, those from social wasps and bees - such as honeybee, hornets and paper wasps - have been well documented. Their venoms are composed of a number of peptides and proteins and used for defending their nests and themselves from predators. In contrast, the venoms of solitary wasps and bees have not been the object of further research. In case of solitary bees, only major peptide components in a few venoms have been addressed. Therefore, the aim of the present study was to explore the peptide component profile of the venom from the solitary bee Xylocopa appendiculata circumvolans by peptidomic analysis with using LC-MS. Methods: A reverse-phase HPLC connected to ESI-OrbiTrap MS was used for LC-MS. On-line mass fingerprinting was made from TIC, and data-dependent tandem mass spectrometry gave MSMS spectra. A major peptide component was isolated by reverse-phase HPLC by conventional way, and its sequence was determined by Edman degradation, which was finally corroborated by solid phase synthesis. Using the synthetic specimen, biological activities (antimicrobial activity, mast cell devaluation, hemolysis, leishmanicidal activity) and pore formation in artificial lipid bilayer were evaluated. Results: On-line mass fingerprinting revealed that the crude venom contained 124 components. MS/MS analysis gave 75 full sequences of the peptide components. Most of these are related to the major and novel peptide, xylopin. Its sequence, GFVALLKKLPLILKHLH-NH2, has characteristic features of linear cationic α-helical peptides; rich in hydrophobic and basic amino acids with no disulfide bond, and accordingly, it can be predicted to adopt an amphipathic α-helix secondary structure. In biological evaluation, xylopin exhibited broad-spectrum antimicrobial activity, and moderate mast cell degranulation and leishmanicidal activities, but showed virtually no hemolytic activity. Additionally, the peptide was able to incorporate pores in artificial lipid bilayers of azolectin, confirming the mechanism of the cytolytic activity by pore formation in biological membranes. Conclusions: LC-ESI-MS and MS/MS analysis of the crude venom extract from a solitary bee Xylocopa appendiculata circumvolans revealed that the component profile of this venom mostly consisted of small peptides. The major peptide components, xylopin and xylopinin, were purified and characterized in a conventional manner. Their chemical and biological characteristics, belonging to linear cationic α-helical peptides, are similar to the known solitary bee venom peptides, melectin and osmin. Pore formation in artificial lipid bilayers was demonstrated for the first time with a solitary bee peptide.

O objetivo do presente estudo foi verificar a atividade do composto maleato de estanho tri-n-butílico contra Staphylococcus aureus e Aspergillus niger, após sua aplicação industrial em 40 amostras de carpetes de diferentes materiais (polipropileno, poliéster, poliamida e lã). Os ensaios qualitativos foram realizados através de duas metodologias: Zona de Inibição (ZI) e Superfície de Inibição (Impressão). Os carpetes tratados com o produto apresentaram 100% de inibição de crescimento bacteriano (Staphylococcus aureus) e fúngico (Aspergillus niger), sob as condições desse estudo. A inibição de crescimento microbiano foi mais elevada na porção superior dos carpetes. As metodologias empregadas parecem ser adequadas para testar a atividade bactericida e fungicida do maleato de estanho tri-n-butílico. A metodologia de impressão confirmou os resultados obtidos no ensaio de zona de inibição. Estudos futuros utilizando as mesmas metodologias são necessários para confirmação destes dados.

The aim of the present study was to verify the activity of the Tri-N-Butyl Tin maleate compound against Staphylococcus aureus and Aspergillus niger, after its industrial application in 40 samples of carpets of different materials (polypropylene, polyester, polyamide and wool). The qualitative assays were performed through two methodologies: Inhibition Halo (HZ) and Inhibition of Surface (Print). The carpet with the product inhibited 100% of bacterial (Staphylococcus aureus) and fungi (Aspergillus niger) growth, under the conditions of this study. The microbial inhibition was higher in upper portion of carpets. The methodologies employed appear to be adequate to test the bactericide and fungicide activities of the Tri-N-Butyl Tin maleate. The print methodology confirmed the results obtained by the inhibition zone assay. Further studies using the same methodologies are needed to confirm our results.

O presente trabalho teve por objetivo avaliar um programa de monitoramento microbiológico ambiental para áreas limpas, ou ambientes classificados, envolvidas na produção, envasamento e controle dos imunobiológicos produzidos pelo Instituto Butantan. Este monitoramento permitiu a quantificação, a caracterização e a sazonalidade da população microbiana presente no ar e nos operadores, e a determinação dos limites de alerta e ação. O número total de bactérias detectáveis revelou diferenças nos sistemas de purificação de ar instalados e o impacto operacional ocasionado pelos procedimentos realizados. A população microbiana característica foi composta por bactérias dos gêneros Staphylococcus sp, Micrococcus sp, Bacillus sp e por fungos filamentosos do gênero Penicillium sp. A maior concentração de microrganismos ocorreu nos períodos de verão e primavera. Os limites internos de alerta e ação estabelecidos asseguram os procedimentos operacionais, recomendando o monitoramento microbiológico ambiental a outros laboratórios envolvidos na produção de vacinas, soros hiperimunes e imunobiológicos.

The aim of the present work was to evaluate an environmental monitoring program for clean rooms, or classified environments, involved in the filling and quality control of biological products produced by Butantan Institute, São Paulo, Brazil. This monitoring established the quantification, characterization and seasonality of the microorganisms in air and operators and, moreover, determined the alert and action limits. The total detectable microbial number showed some contrasts in installed air purification systems and in the operational impact on adopted procedures. The typical microbial population consisted of Staphylococcus sp, Micrococcus sp, Bacillus sp and Penicillium sp. The highest microorganism concentration occurred during summer and springtime. The established internal alert and action limits supported the operational procedures. Therefore, the environmental monitoring program is recommended for other laboratories involved in the production of vaccines, hyperimmune sera and biopharmaceuticals.

We identified different diarrheagenic (DEC) Escherichia coli pathotypes isolated from 1,207 children with and without acute endemic diarrhea in Salvador, Bahia, Brazil collected as part of a case-control study. Since the identification of DEC cannot be based on only biochemical and culture criteria, we used a multiplex polymerase chain reaction developed by combining five specific primer pairs for Enteropathogenic Escherichia coli (EPEC), Shiga toxin-producing E. coli/ Enterohaemorrhagic E. coli (STEC/EHEC), Enterotoxigenic E. coli (ETEC) and Enteroaggregative E. coli (EAEC) to detect these pathotypes simultaneously in a single-step reaction. In order to distinguish typical and atypical EPEC strains, these were tested for the presence of EAF plasmid. The prevalence of diarrheagenic E. coli in this sample of a global case-control study was 25.4% (259 patients) and 18.7% (35 patients) in the diarrhea group (1,020 patients) and the control group (187 patients), respectively. The most frequently isolated pathotype was EAEC (10.7%), followed by atypical EPEC (9.4%), ETEC (3.7%), and STEC (0.6%). Typical EPEC was detected only in one sample. The prevalence of the pathotypes studied in children with diarrhea was not significantly different from that in children without diarrhea.

O objetivo do presente estudo foi verificar a eficácia do acaricida maleato de estanho tri-n-butílico, aplicado industrialmente em amostras de carpetes, tecidos de revestimentos de móveis e de calçados, assim como de espumas de colchão. Aproximadamente 100 ácaros adultos da espécie Dermatophagoides pteronyssinus foram inoculados em placa de Petri contendo a amostra (pedaço de colchão, tecido ou carpete), tratada com o produto acaricida, coletados aleatoriamente. Foi acrescentado sobre a amostra, meio de cultivo para a manutenção dos ácaros. Cada placa foi examinada após 1, 2, 3, 7 e 30 dias de incubação a 25 ºC e 75% de U.R.A. (umidade relativa do ar), sob microscópio estereoscópico com 40X de aumento. O acaricida maleato de estanho tri-n-butílico apresentou 100% de eficácia acaricida após 30 dias da aplicação, em condições ótimas para a manutenção dos ácaros.

The aim of this study was to determine the effectiveness of the acaricide tri-n-butyl tin maleate, industrially applied to samples of carpets, mattress foam, and fabrics used for furniture upholstery, soft toys and shoe uppers. Approximately 100 adult house dust mites of the species Dermatophagoides pteronyssinus were inoculated into a Petri dish containing the sample (a piece of carpet, mattress foam, or fabric), treated with the acaricide, randomly collected. Mite-maintenance culture medium was added on top of each sample. After one, two, three, seven and 30 days of incubation at 25 ºC and 75% relative humidity, each dish was examined using a 40X stereoscopic microscope (40X). One hundred percent acaricide effectiveness was obtained in treated materials by the end of the 30th-day postinoculation period, under optimal conditions for mite maintenance.

We report the frequency of the different diarrheagenic Escherichia coli (DEC) categories isolated from children with acute endemic diarrhea in Salvador, Bahia. The E. coli isolates were investigated by colony blot hibridization whit the following genes probes: eae, EAF, bfpA, Stx1, Stx2, ST-Ih, ST-Ip, LT-I, LT-II, INV, and EAEC, as virulence markers to distinguish typical and atypical EPEC, EHEC/STEC, ETEC, EIEC, and EAEC. Seven of the eight categories of DEC were detected. The most frequently isolated was atypical EPEC (10.1%) followed by ETEC (7.5%), and EAEC (4.2%). EHEC, STEC, EIEC, and typical EPEC were each detected once. The strains of ETEC, EAEC, and atypical EPEC belonged to a wide variety of serotypes. The serotypes of the others categories were O26:H11 (EHEC), O21:H21 (STEC), O142:H34 (typical EPEC), and O?H55 (EIEC). We also present the clinical manifestations and other pathogenic species observed in children with DEC. This is the first report of EHEC and STEC in Salvador, and one of the first in Brazil.

O objetivo do estudo foi identificar ácaro infestando colônia de Aedes aegypti e Ae. albopictus, além da investigação de fonte de contaminação. A espécie de ácaro foi identificada pela montagem, em meio de Hoyer, entre lâmina e lamínula e observados em microscopia óptica, revelando tratar-se de Tyrophagus putrescentiae. Amostras de ração para peixe e de ração para coelho foram mantidas em estufa BOD a 27°C e 90% de umidade, por 21 dias, sendo examinadas semanalmente. A presença de T. putrescentiae foi somente observada em amostras de ração para coelho. Presume-se que T. putrescentiae tenha sido introduzido nas colônias pela forésia em cobaias guinea-pigs, encontrando em ambiente de insetário na ocasião do evento (28,7°C, 72% de umidade e fotoperíodo de 14L:10E horas) condições favoráveis para o seu desenvolvimento.

The present study aimed at identifying a mite infesting a colony of Aedes aegypti and Ae. albopictus as well as investigating the source of infestation. The mite species was identified after it was mounted on slides in Hoyer's medium and examined under an optical microscope. It showed to be Tyrophagus putrescentiae. Fish and rabbit food samples were kept in an environmental chamber at 27ºC and 90% humidity for 21 days and were weekly examined. The presence of T. putrescentiae was only observed in rabbit food. It is assumed that T. putrescentiae had been introduced into the colonies through phoresy among guinea pigs, which were kept at the time near the insectary under favorable conditions for their development (28.7ºC, 72% humidity, photoperiod of 14L:10D hours).

O objetivo do estudo foi identificar ácaro infestando colônia de Aedes aegypti e Ae. albopictus, além da investigação de fonte de contaminação. A espécie de ácaro foi identificada pela montagem, em meio de Hoyer, entre lâmina e lamínula e observados em microscopia óptica, revelando tratar-se de Tyrophagus putrescentiae. Amostras de ração para peixe e de ração para coelho foram mantidas em estufa BOD a 27°C e 90% de umidade, por 21 dias, sendo examinadas semanalmente. A presença de T. putrescentiae foi somente observada em amostras de ração para coelho. Presume-se que T. putrescentiae tenha sido introduzido nas colônias pela forésia em cobaias guinea-pigs, encontrando em ambiente de insetário na ocasião do evento (28,7°C, 72% de umidade e fotoperíodo de 14L:10E horas) condições favoráveis para o seu desenvolvimento.

The present study aimed at identifying a mite infesting a colony of Aedes aegypti and Ae. albopictus as well as investigating the source of infestation. The mite species was identified after it was mounted on slides in Hoyer's medium and examined under an optical microscope. It showed to be Tyrophagus putrescentiae. Fish and rabbit food samples were kept in an environmental chamber at 27ºC and 90% humidity for 21 days and were weekly examined. The presence of T. putrescentiae was only observed in rabbit food. It is assumed that T. putrescentiae had been introduced into the colonies through phoresy among guinea pigs, which were kept at the time near the insectary under favorable conditions for their development (28.7ºC, 72% humidity, photoperiod of 14L:10D hours).