Evaluation of sensitivity to zinc and copper of Diophrys appendiculata(Protozoa, Ciliophora) and their associated bacteria, both isolated from a tropical polluted bay

Ciliates are an essential component of microbial food webs, connecting biomass production to higher trophic levels and providing substrates for bacterial growth. Bacteria are widespread in sea sediment and the first to metabolize organic matter. Heavy metals are toxic and bind to particulate organic matter. This work aims to study the influence of heavy metals (Zn and Cu) on D. appendiculata and their naturally associated bacteria, both from Guanabara Bay during 96 h bioassay with 0, 0.001, 0.009, 0.05, 0.1 mg L-1 and maximum concentration stipulated by CONAMA 357 (Zn 5.0; Cu 1.0 mg L-1). It was analyzed for tolerance, resistance and biomass production. In Zn control, ciliate produced 1.24x102-2.47x103 μg C cm-3 (0-96 h), with 0.009 mg Zn L-1 reaches 2.49x102-1.93x103 μg C cm-3 (0-96 h). Their naturally associated bacteria, in Zn control, produced 5.0x10-2-5.67x10-2 μg C cm-3 (0-96 h), with 0.1 mg Zn L-1 4.87x10-2-8.38x10-2 μg C cm-3 (0-96 h). In Cu control, D. appendiculata produced 1.04x102-3.12x102 μg C cm-3 (0-96 h), with 0.009 mg Cu L-1 biomass was 8.31x101-7.27x10-2 μg C cm-3 (0-96 h) and with CONAMA concentration was below detection level. Their naturally associated bacteria, in Cu control, produced 1.20x10-2-5.14x10-2 μg C cm-3 (0-96 h) and with 0.1 mg Cu L-1 7.40x10-4-3.81x10-2 μg C cm-3 (0-96 h), with CONAMA concentration 5.14x10-2 μg C cm-3 (96 h). D. appendiculata are tolerant to 0.09 mg L-1 and resistant to 0.1 mg L-1 after 24 h to Zn-Cu and LD50 stipulated was 1.17 (Zn) 0.90 (Cu) mg L-1. Microbial loop was functional in low concentration of Zn and Cu, but their diversity in nature was affected.(AU)

Evaluation of sensitivity to zinc and copper of Diophrys appendiculata(Protozoa, Ciliophora) and their associated bacteria, both isolated from a tropical polluted bay

Ciliates are an essential component of microbial food webs, connecting biomass production to higher trophic levels and providing substrates for bacterial growth. Bacteria are widespread in sea sediment and the first to metabolize organic matter. Heavy metals are toxic and bind to particulate organic matter. This work aims to study the influence of heavy metals (Zn and Cu) on D. appendiculata and their naturally associated bacteria, both from Guanabara Bay during 96 h bioassay with 0, 0.001, 0.009, 0.05, 0.1 mg L-1 and maximum concentration stipulated by CONAMA 357 (Zn 5.0; Cu 1.0 mg L-1). It was analyzed for tolerance, resistance and biomass production. In Zn control, ciliate produced 1.24x102-2.47x103 μg C cm-3 (0-96 h), with 0.009 mg Zn L-1 reaches 2.49x102-1.93x103 μg C cm-3 (0-96 h). Their naturally associated bacteria, in Zn control, produced 5.0x10-2-5.67x10-2 μg C cm-3 (0-96 h), with 0.1 mg Zn L-1 4.87x10-2-8.38x10-2 μg C cm-3 (0-96 h). In Cu control, D. appendiculata produced 1.04x102-3.12x102 μg C cm-3 (0-96 h), with 0.009 mg Cu L-1 biomass was 8.31x101-7.27x10-2 μg C cm-3 (0-96 h) and with CONAMA concentration was below detection level. Their naturally associated bacteria, in Cu control, produced 1.20x10-2-5.14x10-2 μg C cm-3 (0-96 h) and with 0.1 mg Cu L-1 7.40x10-4-3.81x10-2 μg C cm-3 (0-96 h), with CONAMA concentration 5.14x10-2 μg C cm-3 (96 h). D. appendiculata are tolerant to 0.09 mg L-1 and resistant to 0.1 mg L-1 after 24 h to Zn-Cu and LD50 stipulated was 1.17 (Zn) 0.90 (Cu) mg L-1. Microbial loop was functional in low concentration of Zn and Cu, but their diversity in nature was affected.

Bacterioplankton features and its relations with doc characteristics and other limnological variables in Paraná river floodplain environments (PR/MS-Brazil)

Since the introduction of the Microbial Loop concept, many studies aimed to explain the role of bacterioplankton and dissolved organic carbon (DOC) in aquatic ecosystems. Paraná River floodplain system is a very complex environment where these subjects were little explored. The aim of this work was to characterize bacterial community in terms of density, biomass and biovolume in some water bodies of this floodplain and to verify its temporal variation and its relation with some limnological variables, including some indicators of DOC quality, obtained through Ultraviolet-visible (UV-VIS) and fluorescence spectroscopic analysis. Bacterial density, biomass and biovolume are similar to those from other freshwater environments and both density and biomass were higher in the period with less rain. The limnological and spectroscopic features that showed any relation with bacterioplankton were the concentrations of N-NH4 and P-PO4, water transparency, and some indicators of DOC quality and origin. The analysis of these relations showed a possible competition between bacterioplankton and phytoplankton for inorganic nutrients and that the DOC used by bacterioplankton is labile and probably from aquatic macrophytes.

Confecção de uma coleção de fungos entomopatogênicos com isolados encontrados em culturas agrícolas e ambientes de preservação ambiental em mato grosso do sul

ABSTRACT With the advance of deforestation, the practice of unplanned farming and the use of toxic pesticides in the environment, it is estimated that the major part of the diversity of entomopathogenic fungi present in the environmental has been disturbed. Preservation in the form of a culture collection is a fundamentally important practice for the maintenance of the diversity of these microorganisms. The present study was aimed at the systematical collection of sample soil and dead and moribund insects that present the extrusion of such pathogens or with characteristics of such diseases in crops or environmental preservation areas. The insects were collected and identified in the laboratory, where the pathogen was isolated in PDA culture medium. The Petri dishes were maintained in a B.O.D. chamber at 25 ± 1° C, 70 ± 10% RH and 12 hours of photophase, for 7 days. After obtaining pure cultures, the conidia were harvested with a metallic loop, stored in plastic Eppendorf tubes, and maintained in a freezer at -4º C. Up to the present, the following entomopathogenic fungi have been isolated: Beauveria bassianaparasitizing Lagria villosa (UFGD 01), Hypothenemus hampei(UFGD 06 and UFGD 11), an insect of the Crisomellidae family (UFGD 02), and from soil (UFGD 16); the fungi Metarhizium anisopliae parasitizing Zulia entreriana (UFGD 05), Mahanarva fimbriolata (UFGD 03), M. posticata (UFGD 07) and from soil (UFGD 12); Paecilomyces fumosoroseus attacking an insect of the Scarabaeidae family (UFGD 04), L. villosa (UFGD 13), and from soil (UFGD 09 and UDGF 15); P. farinosus isolated from soil (UFGD 08), and the fungi Nomuraea rileyi parasitizing the caterpillar Anticarsia gemmatalis (UFGD 14).

RESUMO Com o avanço do desmatamento, a prática de culturas agrícolas não planejadas e a utilização de defensivos agrícolas tóxicos ao meio ambiente, estima-se que grande parte da diversidade de fungos entomopatogênicos presente no meio ambiente esteja sendo alterada. A preservação na forma de coleção de culturas é uma prática de fundamental importância para a manutenção da biodiversidade destes micro-organismos. Este trabalho objetivou a coleta sistematizada de amostras de solo e de insetos mortos e moribundos que apresentassem a extrusão do patógeno ou com características da doença, em culturas agrícolas e área de preservação ambiental. Os insetos coletados foram identificados e levados para o laboratório, onde se isolou o patógeno em meio de cultura BDA. As placas foram mantidas em B.O.D. a 25± 1° C, 70 ± 10% UR e fotofase de 12 horas, durante 7 dias. Após a obtenção de culturas puras, os conídios foram retirados por meio de raspagem com alça metálica, armazenados em tubos plásticos do tipo Eppendorf, e mantidos em freezer a -4º C. Até o momento foram isolados os fungos entomopatogênicos: Beauveria bassiana, parasitando Lagria villosa (UFGD 01), Hypothenemus hampei (UFGD 06 e UFGD 11), um inseto da família Crisomellidae (UFGD 02) e do solo (UFGD 16); o fungo Metarhizium anisopliae parasitando Zulia entreriana (UFGD 05), Mahanarva fimbriolata (UFGD 03), M. posticata (UFGD 07) e do solo (UFGD 12); Paecilomyces fumosoroseus atacando um inseto da família Scarabaeidae (UFGD 04), Lagria villosa (UFGD 13) e do solo (UFGD 09 e 15); P. farinosus isolado a partir do solo (UFGD 08) e o fungo Nomuraea rileyi parasitando a lagarta Anticarsia gemmatalis (UFGD 14).

Evaluation of the Diramic system for urine cultures

The use of the Diramic system in microbiological diagnosis of urinary tract infections (UTI) was evaluated. This system was developed at the National Center for Scientific Research of Cuba, and it reads turbidimetric changes of microbial growth in culture media at 37ºC incubated for 4 hours. A total of 396 urine specimens were tested in the Laboratory of Microbiology of the School of Medicine - UNESP, Brazil using the Diramic system and the counting of colony forming units per urine millimeter (calibrated loop) as the reference method. The coincidence rate between the two methods was 96.46% (382 urine samples), and the differences in results were not significant (p 0.05). Sensitivity and specificity rates were 84.37% and 98.80%, respectively. False negative and false positive rates were 2.50% and 1.01%, respectively. The microorganisms isolated from positive urines were: Escherichia coli (68.75%); Klebsiella pneumoniae (10.94%); yeast (6.25%); Pseudomonas aeruginosa (4.69%); Enterobacter cloacae (3.12%); Proteus mirabilis, coagulase negative Staphylococcus, Morganella morganii, and Citrobacter freundii (1.56% each). The Diramic system was effective as screening method for urine cultures, however restrictions in the UTI diagnosis caused by yeasts and patients undergoing antibiotic therapy were negative characteristics of the system.

O sistema Diramic foi avaliado para o diagnóstico das infecções do trato urinário (ITU). O sistema Diramic foi desenvolvido em Cuba e possibilita resultados de diagnóstico das infecções do trato urinário (ITU) em quatro horas e baseia-se na variação da turvação do crescimento microbiano no meio de cultura após incubação a 37ºC/4 horas. 396 amostras de urinas provenientes de ambulatórios e enfermarias do HC da FMB-UNESP-Botucatu/SP foram analisadas pelo sistema Diramic. O método da alça calibrada (AC) foi adotado como método de referência. A taxa de coincidência entre os dois métodos foi de 96,46% (382 amostras de urina), não havendo diferença significativa entre os resultados obtidos nos dois métodos. Os resultados para sensibilidade e especificidade foram 84,37 e 98,80% respectivamente e 10 resultados no Diramic foram falsos negativos (2,5%) e 4 falso positivos (1,01%). Os microrganismos identificados nas urinas positivas foram Escherichia coli (68,75%), Klebsiella pneumoniae (10,94%), leveduras (6,25%), Pseudomonas aeruginosa (4,69%), Enterobacter cloacae (3,12%) e Proteus mirabilis, Staphyloccocus coagulase negativo, Morganella morganii e Citrobacter freundii também foram identificadas (1,56% para cada espécie). O método Diramic foi eficiente na triagem das urinoculturas, porém verificou-se algumas restrições quanto ao diagnóstico das infecções do trato urinário quando causadas por leveduras e em pacientes submetidos a antibioticoterapia.