Determinação da atividade antimicrobiana dos extrativos das folhas/galhos, pericarpo e sementes da Crotalaria pallida (Leguminosae)

O estudo de novas alternativas terapêuticas no tratamento de infecções microbianas tornou-se crescente no cenário científico devido à grande variação genética desses micro-organismos, que resultou na resistência aos antimicrobianos existentes. A grande diversidade na flora brasileira e a ampla utilização das plantas como medicamentos pela população justificam os estudos e o crescente interesse pela descoberta de novos compostos bioativos isolados dos vegetais. Plantas usualmente utilizadas na agricultura, apenas como adubação verde, por exemplo, são atualmente alvo de estudos científicos com potenciais promissores de atuação como de produtos terapêuticos. Plantas da família Leguminosae, amplamente conhecidas e utilizadas como fornecedoras de nitrogênio ao solo, vem sendo estudadas por diversas áreas da saúde para comprovarem a ação de compostos isolados entre estes os alcalóides pirrolidizínicos como antiinflamatórios, antibióticos e até como veneno para pragas. O objetivo do presente estudo foi determinar, a partir de extrativos de Crotalaria pallida, a atividade antimicrobiana in vitro utilizando cepas padrões de: Staphylococccus aureus, Escherichia coli, Salmonella sp e da levedura Candida albicans. Para determinação dessa atividade foi utilizada a técnica de diluição em microplaca que possibilitou o estudo da atividade do extrativo vegetal e da concentração inibitória mínima, isto é, concentração bactericida e/ou bacteriostática mínima e concentração fungiostática e/ou fungicida mínima. A Crotalaria pallida, não apresentou atividade frente aos micro-organismos testados nas condições padronizadas neste estudo

Taxonomic and distributional notes on Telebasis Selys, 1865, with a redescription of T. pallida Machado, 2010, and an evaluation of the T. racenisi Bick & Bick, 1995 "complex" of species (Odonata, Coenagrionidae)

A full checklist of the species of Telebasis Selys, 1865, housed in the Brazilian collections Colecao Entomologica Prof. Jose Alfredo Pinheiro Dutra, Departamento de Zoologia, Instituto de Biologia, Universidade Federal do Rio do Janeiro (DZRJ), and Museu de Zoologia, Universidade de Sao Paulo (MZSP) is presented. A total of 325 specimens representing 19 species were studied. Ten new records for Brazilian States were found for T. carmesina Calvert, 1909 (Rio de Janeiro and Rio Grande do Sul), T. corallina (Selys, 1876) (Pernambuco), T. demarara (Williamson, 1917) (Maranhao), T. filiola (Perty, 1834) (Paraiba and Santa Catarina), T. gigantea Daigle, 2002 (Sao Paulo), T. inalata (Calvert, 1961) (Mato Grosso do Sul), T. pallida Machado, 2010 (Goias) and T. obsoleta (Selys, 1876) (Mato Grosso do Sul), as well as a new record of T. carminita Calvert, 1909 for Suriname. Telebasis pallida Machado, 2010 is redescribed and diagnosed based on 14 males collected near the type locality, and its genital ligula is described and illustrated for the first time. Furthermore, the status of the three species of the Telebasis racenisi Bick & Bick, 1995 complex is evaluated. Of these, Telebasis pareci Machado, 2010 syn. n. is proposed as junior subjective synonym of Telebasis lenkoi Machado, 2010, and a possible synonymy among the three species is discussed under T. racenisi. ((c) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Interspecific competition between the potato cyst nematode species Globodera pallida and G. rostochiensis

The two potato cyst nematode species, Globodera pallida and G. rostochiensis, are among the most important pests of potato. PCN are difficult to manage, while the two species respond differently to the main control methods. An increase in the incidence of G. pallida had been reported and is generally attributed to greater effectiveness of control measures against G. rostochiensis. The status of PCN in Ireland was studied using PCR. The results demonstrated qPCR to be an efficient means of high-throughput PCN sampling, being able to accurately identify both species in mixed-species populations. Species discrimination using qPCR revealed an increase in the incidence of G. pallida in Ireland in the absence of G. pallida-selective control measures. The population dynamics of G. pallida and G. rostochiensis in Ireland were studied in mixed- and single-species competition assays in vivo. G. pallida proved to be the more successful species, with greater multiplication in mixed- than single-species populations, with G. rostochiensis showing the opposite. This effect was similarly observed in staggered inoculation trials and population proportion trials. It was hypothesised that the greater G. pallida competitiveness could be attributed to its later hatch. G. pallida exhibited a later peak in hatching activity and more prolonged hatch, relative to G. rostochiensis. G. rostochiensis hatch was significantly reduced in mixedspecies hatching assays. G. pallida hatch was significantly higher when hatch was induced in potato root leachates containing G. rostochiensis-specific compounds, indicating that G. pallida hatch is stimulated upon perception of G. rostochiensis–derived compounds. Rhizotron studies revealed that root damage, caused by feeding of the early-hatching G. rostochiensis, resulted in increased lateral root proliferation and significantly increased G. pallida multiplication. Split-root trials indicated a significant G. pallida-induced ISR effect. G. rostochiensis multiplication was significantly reduced in split-root rhizotrons when G. pallida colonised roots before or after G. rostochiensis infection.

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

GOMES, Carlos E. M. et al. Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly). Plant Physiology and Biochemistry (Paris), v. 43, n. 12, p. 1095-1102, 2005.ISSN 0981-9428. DOI:10.1016/j.plaphy.2005.11.004.

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

GOMES, Carlos E. M. et al. Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly). Plant Physiology and Biochemistry (Paris), v. 43, n. 12, p. 1095-1102, 2005.ISSN 0981-9428. DOI:10.1016/j.plaphy.2005.11.004.

Purificação, caracterização e atividade bioinseticida de um inibidor de tripsina de sementes de Crotalaria pallida

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulphate fractionation, affinity chromatography on immobilized Trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named ITC, had Mr of 32.5 kDa by SDS-PAGE and was composed by two subunits with 27.7 and 5.6 kDa linked by disulphide bridges, a typical characteristic of Kunitz-Inhibitor family. ITC was stable until 50°C, and at 100°C its residual activity was of about 60%. Also, ITC was stable at pHs 2 to 12. The inhibition of trypsin by ITC was non-competitive, with a Ki of 8,8 x 10-7M. ITC inhibits weakly other serine proteinases such as chymotrypsin and elastase. The inhibition of papain (44% of inhibition), a cysteine proteinase was an indicative of the bi-functionality of ITC. In vitro assays against digestive proteinases from several Lepdoptera, Diptera and Coleoptera pests were made. ITC inhibited in 100% digestive enzymes of Ceratitis capitata (fruit fly), Spodoptera frugiperda and Alabama argillacea, the last one being a cotton pest. It also inhibited in 74.4% Callosobruchus maculatus (bean weevil) digestive enzymes, a Coleoptera pest. ITC, when added in artificial diet models, affected weakly the development of C. capitata larvae and it had a WD50 of 2.65% to C. maculatus larvae

Bioensaios de toxicidade genética com plantas superiores Tradescantia (MCN e SHM), milho e soja.

Para se evitarem agentes mutagênicos no ambiente são necessários indicadores sensíveis para detectar todo o espectro desses compostos. Tais sistemas indicadores tem sido definidos como testes de mutagênese de tipo II, ou seja, aqueles que apresentam alta sensibilidade e baixa especificidade. A maioria dos bioensaios vegetais são considerados testes do tipo II, com especial referencia para os ensaios do micronúcleo e do pelo estaminal em Tradescantia (comelinacea), e o ensaio do grão de pólen ceroso em milho. Outro bioensaio vegetal de interesse para o monitoramento de agentes mutagênicos ambientais e o teste do mosaicismo em soja, que permite especulação sobre o mecanismo envolvido na toxicidade genética. O bioensaio não exige qualquer instrumentação sofisticada, e e adequado para experimentação in situ. Esses bioensaios vem sendo empregados com sucesso em estudos sobre a mutagenicidade de pesticidas aplicados conforme prescrição agronômica. Resultados adicionais sobre a utilização desses bioensaios in situ nas mais diversas situações indicam que esses testes são adequados para o monitoramento extensivo de mutagênese ambiental.

High performance liquid chromatography determined alkamide levels in Australian-grown Echinacea spp.

Extracts of Echinacea spp. are widely used as therapeutic immunostimulants with such activity being attributed in part to the alkamide fractions of these plants. Using high performance liquid chromatography, the levels of 8 alkamides, including 2 tetraene alkamides (dodeca-2E, 4E, 8Z, 10E/Z-tetraenoic acid isobutylamide), were quantitatively determined in 2 Australian-grown Echinacea spp. Overall, the levels of alkamides in Australian-grown E. angustifolia were found to be comparable with levels obtained in this study and other studies for USA and European Echinacea spp. However, results obtained for one sample of E. angustifolia suggested that it may have been mislabelled and that it was most likely a sample of E. pallida. Levels of tetraene alkamides in Australian-grown E. purpurea were also similar to, if not higher, than levels which have been reported for the same species grown in Germany and the USA. Preliminary studies on the stability of alkamide compounds in E. angustifolia indicated that they are susceptible to degradation, with a 13% loss of alkamide level over 2 months. Overall, results indicate that there is considerable potential to develop Echinacea as a viable crop in Australia.

Responses of two species of mesquite to initial and follow-up applications of selected herbicides in a potted trial

Quilpie mesquite (Prosopis velutina) is an invasive woody weed that is believed to have been introduced into south-west Queensland in the 1930s. Following the withdrawal of 2,4,5-T, research on P. pallida resulted in revised recommendations for control of all Prosopis spp. in Queensland. Adoption of many of these recommendations for Quilpie mesquite control produced substandard results. Following a pilot trial, a shade-house experiment was conducted to determine the differences in susceptibility of two species of mesquite, P. velutina and P. pallida, to commonly available herbicides. It was hypothesized that P. velutina was less susceptible than P. pallida, based upon claims that the registered chemical recommendations for Prosopis spp. were not sufficiently effective on P. velutina. Nine foliar herbicide treatments were applied to potted shade-house plants. Treatment effects indicated differing susceptibility between the two species. P. velutina consistently showed less response to metsulfuron, fluroxypyr, 2,4-D/picloram and triclopyr/picloram, compared to the glyphosate formulations, where negligible differences occurred between the two species. The response to glyphosate was poor at all rates in this experiment. Re-application of herbicides to surviving plants indicated that susceptibility can decrease when follow-up application is in autumn and the time since initial application is short. The relationship between leaf structure and the volume of spray adhering to a plant was assessed across species. The herbicide captured by similar-sized plants of each species differed, with P. pallida retaining a greater volume of herbicide.

Some freshwater ostracods (Crustacea: Ostracoda) from South Wales

As background to a study of the application of astracods in environmental archaeology, a number of sites in South Wales were visited and sampled. Sites included seven broad environmental categories consisting of lakes, permanent ponds, non-permanent ponds, semi-static canals and reens (drainage ditches), non-permanent small lotic water-bodies, permanent fast-flowing waters and wells. In all, twenty-three species were recorded, and with one exception all belonged to the predominately freshwater Cypridoidea. Overall the most commonly encountered species in South Wales was Cypria ophthalmica. Comparing finds with earlier records, it would appear that Ilyocypris bradyi, Candona pratensis, Eucypris lilljeborgi, Herpetocypris chevreuxi, Potamocypris variegata, P. similis and P. pallida are new additions to the Welsh fauna.

Biopotentials of marine sponges from China oceans: past and future

An extensive literature survey of over 17 Journals was carried out on Chinese sponges and their natural products in the period from 1980 to 2001. This review is thus intended to provide the first thorough overview of research on marine sponges from China Ocean territories. Information is provided about the rather-limited taxonomic study of Chinese marine sponges, with an analysis on their distribution and diversity. Research findings on the natural products and their bioactivity screening from Chinese sponges are summarized. The weaknesses, gaps and problems in the past R&D program of Chinese sponges are identified, which point to the future opportunities in exploiting these huge untapped sponge resources. The report is expected to serve as an entry point for understanding Chinese sponges and for furthering R&D on their bioactive compounds for new drug development. (C) 2003 Elsevier Science B.V. All rights reserved.

微型浮游动物及其对浮游植物的摄食压力

本论文研究了渤海，胶州湾和东海20-200 μm微型浮游动物（桡足类幼虫、纤毛虫）的数量和生物量（1% Lugol's 试剂固定1000ml，Utermohl法150倍显微镜检查），在渤海和东海用稀释法（Landry and Hassett, 1982; Burkill et al., 1990）估计了微型浮游动物（<200 μm）对浮游植物的摄食压力。对上述海区调查的时间如下：渤海：1997年6，1998年6月（莱州湾），1998年10月；胶州湾1997年8、11月，1998年2、4、8、11月，1999年2月；东海：1998年7月。结果如下：渤海：1997年6月，只发现一种砂壳纤毛虫，数量为0-980个/L。1998年6月（莱州湾），微型浮游动物数量为30-2390个/L，生物量为1.5-25 μg C/L，水体中生物量为0.1-53.2 mgC m~(-2)。共有13种砂壳纤毛虫和一种无壳纤毛虫。纤毛虫的数量为0-2380个/L，生物量为0-24.52 μg C/L。纤毛虫优势种有无壳纤毛虫、Tintinnopsis pallida、T. amoyensis和T. chinglanensis。微型浮游动物的各个类群呈斑块分布。1998年10月，微型浮游动物的数量为0-770个/L，生物量为0-12.3 μg C/L，水体中生物量为0-136 mgC m~(-2)。在两次大面调查间隔的6天内，发生大风。两次大面站调查海区的温盐分布格局没有很大变化，但微型浮游动物的变化却十分显著，这也反映了恶劣天气对海洋生物的影响。在几个连续站对微型浮游动物的连续监测表明，微型浮游动物的分布与水团有很大关系。胶州湾：桡足类幼虫在8月数量最大（850个/L）。共记录两种无壳纤毛虫和7种砂壳纤毛虫。微型浮游动物数量弯化很大，从10-22600个/L。微型浮游动物的分布格局可能是湾内和湾外海水混合的结果：1997年11月，无壳纤毛虫No.12从湾外涌入，其他航次无壳纤毛虫No.11在湾内发生，向湾外扩散。东海：微型浮游动物数量较少，最大数量为1970个/L，绝大多数站位低于300个/L，生物量为0.08-45 μg C/L，水体50m积分生物量为5-283mgC m~(-2)。微型浮游动物有明显的斑块分布，连续监测没有发现明显的变化。根据稀释培养的结果，微型浮游动物对浮游植物摄食压力如下所述：渤海三个航次的微型浮游动物对浮游植物摄食率为0.13-1.01d~(-1)，每天摄食浮游植物生物量（现存量）的12-49%，初级生产力的34-267%。东海稀释培养的结果为微型浮游动物地浮游植物的摄食率为0.2-1.2d~(-1)，每天摄食浮游动物生物量的20-71%，摄食初级生产力的54-98%。由于稀释培养实验的设计和操作有许多争议之外，本文只将结果列出，对方法不作讨论。

Summertime ciliate and copepod nauplii distributions and micro-zooplankton herbivorous activity in the Laizhou Bay, Bohai Sea, China

The abundance and biomass of ciliated protozoa and copepod nauplii were investigated at 21 grid stations and two anchored stations in the Laizhou Bay, Bohai Sea, China in June 1998. Dilution incubations were carried out to investigate micro-zooplankton grazing pressure at the anchored stations during spring tide and neap tide. The dominant species were Tintinnopsis amoyensis, T. chinglanensis, T. pallida and aloricate ciliates. A total of 13 species of tintinnids were found. The total abundance of ciliates and nauplii ranged from 30 to 2390 ind l(-1) at grid stations. Tintinnopsis amoyensis was the only ciliate found at the anchored stations and in concentrations which varied from 0 to 6700 ind l(-1). The spatial distribution of ciliates was patchy. Tintinnopsis amoyensis and T. pallida were distributed in the Weihe River mouth and Xiaoqinghe River mouth respectively. The aloricate ciliates, T. chinglanensis and Codonellopsis ostenfeldi dominated offshore in sequence. The water mixing process may affect the spatial pattern of the dominant ciliate species. The abundance and biomass of copepod nauplii were in the range of 0-140 ind l(-1) and 0-7 mu g C l(-1) respectively, with the peak appearing at grid station 15. The total biomass of ciliates and copepod nauplii was in the range of 1(.)5-25 mu g C l(-1). Water column biomass of ciliates and nauplii varied from 2(.)37 to 52(.)3 mg C m(-2). At the anchored stations, the phytoplankton growth rates ranged from undetectable to 0 21 d(-1) and micro-zooplankton grazing rates from 0 13 to 0(.)57 d(-1). The grazing pressure of micro-zooplankton were 12 to 43% of the chlorophyll standing stock and 84 to 267% of the chlorophyll (C) 2000 Academic Press.

Novel bioassays to examine the host-finding ability of plant-parasitic nematodes

We present two novel bioassays to be used in the examination of plant-parasitic nematode host-finding ability. The host-finding 'pipette-bulb assay' was constructed from modelled Pasteur pipette bulbs and connecting barrels using parafilm fastenings. This assay examines the direction of second-stage juvenile (J2) migration in response to a host seedling, through a moistened sand substrate, which underlies terminal upward-facing 'seedling bulbs', one containing a host seedling in potting compost, the other with only potting compost. An equal watering regime through both upward-facing seedling bulbs creates a directional concentration gradient of host diffusate chemotactic factors. Positive chemotactic stimuli cause the J2 to orientate and migrate towards the host plant. We present validation data collected from assays of the root-knot nematode, Meloidogyne incognita, and the potato cyst nematode, Globodera pallida, which indicate a highly significant positive attraction of J2 of both species to respective host plants. This represents a simple, quick and inexpensive method of assessing host-finding behaviour in the laboratory. We consider that the pipette-bulb assay improves on previous host-finding/chemo-attraction assays through creating a more biologically relevant environment for experimental J2; analysis is quick and easy, allowing the straightforward interpretation of results. In addition, we have developed an 'agar trough' sensory assay variant which we believe can be used rapidly to ratify nematode responses to chemical gustatory or olfactory cues. This was constructed from a water agar substrate such that two counting wells were connected by a raised central trough, all flooded with water. Two small water agar plugs were dehydrated briefly in an oven and then hydrated in either an attractant, repellent or water control; these plugs were then placed in the terminal counting wells and subsequently leached the attractant or repellent to form a concentration gradient along the central trough, which contained the initial J2 innoculum. Our data show that both M. incognita and G. pallida J2 are positively attracted to host diffusates. In addition, they displayed a strong repulsion in response to 1 M NaCl2. J2 of M. incognita displayed a mild aversion to a non-host oak root diffusate, whereas G. pallida J2 displayed a strong aversion to the same non-host diffusate; neither species responded to a compost leachate. We believe that the agar trough assay improves on previous methods by facilitating rapid diffusion of attractant or repellents. Both of the aforementioned assays were designed as tools to assess the impact of RNAi-based reverse genetics screens for gene targets involved in chemosensory orientation.