Desenvolvimento de formulações probióticas de isolados clínicos de Lactobacillus spp. para controle de Candida albicans / Development of probiotic formulations of clinical isolates of Lactobacillus spp. for the control of Candida albicans

O uso de probióticos tem sido considerado uma alternativa para o controle das candidoses bucais, como a estomatite protética. Recentemente, nosso grupo de pesquisa isolou cepas clínicas bucais de Lactobacillus rhamnosus (5.2) e Lactobacillus fermentum (20.4) com atividade antifúngica contra Candida albicans. Entretanto, é necessário o desenvolvimento de formulações para uso clínico dessas cepas probióticas na cavidade bucal. Assim, o objetivo desse projeto foi desenvolver formulações probióticas em hidrogel gellan gum com atividade antifúngica que possam ser usadas no controle da estomatite protética. Para o preparo das formulações probióticas, as cepas L. rhamnosus 5.2 ou L. fermentum 20.4 foram incorporadas em formulações com diferentes concentrações de gellan gum (0,6 a 1%). A viabilidade das células de Lactobacillus nas formulações e o sistema de liberação foram testados em diferentes condições de armazenamento, incluindo tempo e temperaturas de estocagem. A seguir, os efeitos das formulações probióticas foram avaliados sobre C. albicans pelo método de difusão em ágar e sobre biofilmes microcosmos por contagem de células viáveis e análise de microscopia eletrônica de varredura. Os resultados demonstraram que todas as concentrações do gellan foram capazes de manter a viabilidade das células de L. rhamnosus e L. fermentum em torno de 5 a 7 log, durante 14 semanas, independentemente da temperatura de armazenamento (4°C ou temperatura ambiente). No teste de difusão em ágar, verificou-se que as formulações probióticas levaram à inibição de cepas padrão e clínicas de C. albicans, demonstrando que as cepas de Lactobacillus incorporadas no gellan mantiveram sua atividade antifúngica. As formulações probióticas também conseguiram inibir os biofilmes microcosmos provenientes de pacientes com estomatite protética, reduzindo o número de células viáveis de Candida e desestruturando a arquitetura dos biofilmes. Concluiu-se que o gellan gum mostrou ser um biomaterial promissor como carreador das cepas probióticas de L. rhamnosus 5.2 e L. fermentum 20.4. Além disso, as formulações probióticas desenvolvidas apresentaram ação antifúngica sobre células de C. albicans e biofilmes de estomatite protética, podendo desta forma atuar na prevenção da candidose bucal. (AU)

The use of probiotics has been considered an alternative for the control of oral candidiasis such as denture stomatitis. Recently, our research group isolated oral clinical strains of Lactobacillus rhamnosus (5.2) and Lactobacillus fermentum (20.4) with antifungal activity against Candida albicans. However, it is necessary to develop formulations for clinical use of these probiotic strains in the oral cavity. Thus, the purpose of this project was to develop probiotic formulations in gellan gum hydrogel with antifungal activity that could be used in the control of denture stomatitis. For the preparation of probiotic formulations, the strains L. rhamnosus 5.2 or L. fermentum 20.4 were incorporated into formulations with different concentrations of gellan gum (0.6 to 1%). The viability of Lactobacillus cells in the formulations and the delivery system were tested under different storage conditions, including storage time and temperatures. Next, the effects of the probiotic formulations were evaluated on C. albicans by the agar diffusion method and on microcosm biofilms by viable cell count and scanning electron microscopy analysis. The results showed that all concentrations of gellan were able to maintain the viability of L. rhamnosus and L. fermentum cells around 5 to 7 log for 14 weeks, regardless of storage temperature (4°C or room temperature). In the agar diffusion test, it was found that the probiotic formulations led to the inhibition of standard and clinical strains of C. albicans, demonstrating that the Lactobacillus strains incorporated into gellan maintained their antifungal activity. Probiotic formulations were also able to inhibit microcosm biofilms from patients with denture stomatitis, reducing the number of viable Candida cells and disrupting the architecture of biofilms. As a conclusion, the gellan gum proved to be a promising biomaterial as a carrier of the probiotic strains of L. rhamnosus 5.2 and L. fermentum 20.4. In addition, the probiotic formulations developed showed antifungal action on C. albicans cells and denture stomatitis biofilms, thus being able to act in the prevention of oral candidiasis. (AU)

Novas estratégias terapêuticas para o controle de biofilmes orais: terapia fotodinâmica e sistema carreador de antimicrobianos / New therapeutic strategics for the control of oral biofilms: photodynamic therapy and drug delivery system

Os biofilmes orais possuem grande relevância clínica por estarem associados com o desenvolvimento de cárie dentária e candidose bucal, que são doenças infecciosas frequentemente encontradas na população. O presente trabalho foi dividido em dois estudos: Estudo 1 que teve como objetivo analisar os efeitos da terapia fotodinâmica antimicrobiana (TFDa), mediada por Fotoenticine (FTC) e Azul de Metileno (AM), sobre biofilmes microcosmos de cárie dentária; e Estudo 2 cujo objetivo foi avaliar o gellan gum como biomaterial para carreador do antifúngico Ester fenetil do ácido caféico (CAPE) contra Candida albicans. No estudo 1, amostras de dentina cariada foram coletadas de diferentes pacientes para formar biofilmes microcosmos in vitro. Os biofilmes foram tratados com FTC ou AM associado à irradiação LED a 660 nm (28,5 J/cm²). Os dados foram analisados pela contagem de Unidades Formadoras de Colônias (UFC/mL). Além disso, a biomassa, estrutura do biofilme e produção de ácidos pelos microrganismos foram determinadas por análises microscópicas ou espectrofotométricas. Os biofilmes de diferentes pacientes apresentaram variações na composição microbiana, sendo formados por estreptococos, lactobacilos e leveduras. No geral, a TFDa diminuiu 3,7 Log10 do total de microrganismos, 2,8 Log10 de estreptococos, 3,2 Log10 de lactobacilos e 3,2 Log10 de leveduras, e atingiu a erradicação de estreptococos do grupo mutans. A TFDa também foi capaz de reduzir a biomassa, desagregar os biofilmes e diminuir a concentração de ácidos em 1,1 a 1,9 mmol de lactato/L. Em relação ao estudo 2, inicialmente, foram preparadas formulações do CAPE em diferentes concentrações de gellan gum (0,6 a 1%). As formulações foram avaliadas em relação ao sistema de liberação e ação antifúngica contra C. albicans. Verificou-se que concentrações mais altas de gellan (0,9 e 1%) levaram a uma liberação mais prolongada do CAPE em relação as concentrações mais baixas. Os valores de concentração inibitória mínima do CAPE sobre C. albicans foram aumentados quando esse composto foi incorporado no gellan. As formulações de CAPE em gellan apresentaram atividade antifúngica tanto em culturas planctônicas como em biofilmes de C. albicans, sendo esses efeitos dependentes do tempo de tratamento. O CAPE e suas formulações em gellan também levaram a uma diminuição da atividade proteolítica de C. albicans. Concluiu-se que a TFDa mediada por Fotoenticine e o sistema carreador de gellan gum podem ser estratégias terapêuticas promissoras para o controle dos biofilmes na cavidade bucal, podendo ser usadas respectivamente no tratamento da cárie e candidose. (AU)

Dental caries and oral candidiasis are infectious diseases frequently found in the population. The present work is divided into two studies, study 1 time as objective: To analyze the effects of antimicrobial photodynamic therapy (aPDT), mediated by Fotoenticine (FTC) and Methylene Blue (MB), on dental caries microcosm biofilms. In study 2, the objective was to evaluate gellan gum as a biomaterial to carry the antifungal caffeic acid phenethyl ester (CAPE) on Candida albicans. To conduct study 1, carious dentin samples were collected from different patients to form in vitro microcosm biofilms. The biofilms were treated with FTC or MB associated with 660 nm red LED irradiation, with energy dose of 28.5 J/cm² and power dose of 40 mW/cm². The data were analyzed by the count of Colony Forming Units (CFU/mL). In addition, the biomass, biofilm structure and acid production of the microorganisms were determined by microscopic or spectrophotometric analysis. The biofilms from different patients showed variations in microbial composition, being formed by streptococci, lactobacilli, and yeasts. Overall, aPDT decreased 3.7 Log10 of total microorganisms, 2.8 Log10 of streptococci, 3.2 Log10 of lactobacilli and 3.2 Log10 of yeasts, and achieved eradication of mutans group streptococci. PDTa was also able to reduce biomass, disaggregate biofilms, and decrease acid concentration by 1.1 to 1.9 mmol lactate/L. For study 2 of this, first the standards of CAPE were determined, such as minimum inhibitory concentration, and absorption peak, then CAPE was incorporated into gellan gum, and then the standard curve test and analysis of CAPE release was performed, finally the formulations were tested on planktonic culture and biofilm of different strains of C. albicans, it was also analyzed the action of this drug on the production of Sap. The MIC found varied from 32 to 64 µg/mL, the release tests showed a gradual release in the higher formulations, finally in the CFU/mL count both in planktonic culture and biofilm the formulations were able to inhibit the fungus. With this it is concluded that both aPDT for oral microcosm and gellan gum as caregiver of CAPE for Candida albicans inhibition are promising. (AU)

Influence of caries activity and number of saliva donors: mineral and microbiological responses in a microcosm biofilm model

Abstract Objective this study evaluated the mineral and microbiological response of biofilms originating from different types of saliva inoculum with distinct levels of caries activity. Methodology the biofilms grown over enamel specimens originated from saliva collected from a single donor or five donors with two distinct levels of caries activity (caries-active and caries-free) or from pooling saliva from ten donors (five caries-active and five caries-free). The percentage surface hardness change (%SHC) and microbiological counts served as outcome variables. Results the caries activity of donors did not affect the %SHC values. Inoculum from five donors compared to a single donor showed higher %SHC values (p=0.019). Higher lactobacilli counts were observed when saliva from caries-active donors was used as the inoculum (p=0.017). Pooled saliva from both caries activity levels showed higher mutans streptococci counts (p<0.017). Conclusion Overall, pooled saliva increased the mineral response of the derived biofilms, but all the inoculum conditions formed cariogenic biofilms and caries lesions independently of caries activity.

Antibacterial dental adhesive containing nanoantibacterial inorganic fillers / 华西口腔医学杂志

OBJECTIVE@#This investigation aimed to develop a novel antibacterial dental adhesive containing nanoantibacterial inorganic fillers and measure the dentin bonding strength, mechanical properties, and antibacterial property of the novel adhesive in vitro.@*METHODS@#Novel nanoantibacterial inorganic fillers containing quaternary ammonium salt with long chain alkyl were synthesized on the basis of previous research. These novel nanoantibacterial inorganic fillers were added into the dental adhesive to prepare novel nanoantibacterial dental resin composite at mass fractions of 0%, 2.5%, 5.0%, 7.5%, and 10%; 0% was used as control. Dentin shear bonding test was used to evaluate the bonding strength. Flexural test was utilized to measure the novel resin composite flexural strength and elastic modulus. A dental plaque microcosm biofilm model with human saliva as inoculum was formed. Colony forming unit, lactic acid production, and live/dead assay of the biofilm on novel dental adhesive were calculated to assess the effect of novel dental adhesive on human dental plaque microcosm biofilm.@*RESULTS@#The dentin shear bond strength, flexural strength, and elastic modulus were 28.9 MPa, 86.6 MPa, and 4.2 GPa, respectively, when the nanoantibacterial inorganic filler mass fraction in the dental adhesive reached approximately 5.0%. Consequently, the dentin shear bond strength and mechanical properties significantly increased. Addition of 2.5% nanoantibacterial inorganic fillers into the dental adhesive exerted no adverse effect on the mechanical properties significantly (P>0.05). Dental adhesive containing 5% or more nanoantibacterial inorganic fillers inhibited the metabolic activity of the dental plaque microcosm biofilm significantly, thereby displaying a strong antibacterial potency (P<0.05).@*CONCLUSIONS@#This novel antibacterial dental adhesive, which contained 5.0% nanoantibacterial inorganic filler, exhibited promising bonding strength, mechanical property, and antibacterial ability. Hence, this adhesive can be potentially used in caries inhibition in dental application.

Effect of Myracrodruon urundeuva and Qualea grandiflora extracts on viability and activity of microcosm biofilm and prevention of enamel demineralization in vitro / Efeito de extratos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade e atividade de biofilme microcosmo e na prevenção da desmineralização do esmalte in vitro

The objective of this study was to evaluate the antimicrobial and anti-caries effects of two plant extracts. The first chapter dealt with a review of the literature whose objective was to discuss the antimicrobial potential of Brazilian natural agents on the biofilm related to dental caries and gingivitis/periodontal disease. The research of the articles was carried out using PubMed. We found a total of 23 papers. Most of the studies were performed using planktonic microorganisms or under clinical trials. Nineteen articles were focused on cariogenic bacteria. From these nineteen articles, eleven were also about periodontopathogenic bacteria. Four studies addressed only periodontopathogenic bacteria. The most tested Brazilian natural agents were green propolis, essential oils of Lippia sidoides and Copaifera sp. Most of the tested agents showed similar results when compared to positive control (essential oils and extracts) or better effect than negative control (green propolis). More studies involving protocols closer to the clinical condition and the use of response variables that allows understanding the mechanism of action of natural agents are necessary before the incorporation of these natural agents into dental products. The second chapter aimed to test the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability of the microcosm biofilm and on the prevention of enamel demineralization. The microcosm biofilm was produced on bovine enamel, using human saliva pool mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva at 100, 10 and 0.1 µg/ml and Q. grandiflora at 100 and 0.1 µg/ml reduced cell viability similarly to the positive control and significantly more than negative control. M. urundeuva at 1000, 100 and 0.1 µg/ml were able to reduce the counting formation unit-CFU counting of lactobacilli sp. and Streptococcus mutans, while Q. grandiflora at 1000 and 1.0 µg/ml significantly reduced the S. mutans CFU counting. On the other hand, the natural extracts did not reduce the production of extracellular polyssacharides, lactic acid and the development of enamel caries lesions. The third chapter aimed to evaluate the effect of hydroalcoholic extracts of M. urundeuva and Q. grandiflora (alone or combined) on the viability of S. mutans biofilm and the prevention of enamel demineralization. S. mutans strain (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration and minimum biofilm eradication concentration were determined to choose the concentrations to be tested under the biofilm model. S. mutans biofilm (5x105 CFU/ml) was produced on bovine enamel using McBain saliva with 0.2% sucrose for 3 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva (isolated or combined) at concentrations equal or higher than 0.625 mg/ml was able to reduce the bacteria viability, whereas Q. grandiflora extract alone showed antimicrobial effect at 5 mg/ml only (p<0.05). On the other hand, none of the extracts was able to reduce the development of enamel caries lesions. Despite the tested natural extracts have antimicrobial effect; they are unable to prevent caries in enamel.(AU)

O objetivo foi avaliar os efeitos antimicrobiano e anti-cárie de dois extratos de plantas. O primeiro capítulo se referiu a uma revisão da literatura cujo objetivo foi discutir o potencial antimicrobiano dos agentes naturais brasileiros sobre o biofilme relacionado à cárie dentária e à gengivite/doença periodontal. A pesquisa dos artigos foi realizada usando o PubMed. Foram encontrados 23 trabalhos. A maioria dos estudos foi realizada utilizando microorganismos na fase planctônica ou ensaios clínicos. Dezenove artigos foram focados em bactérias cariogênicas. Dos dezenove artigos, onze também eram sobre bactérias periodontopatogênicas. Quatro estudos abordaram apenas bactérias periodontopatogênicas. Os agentes naturais brasileiros mais testados foram própolis verde, óleos essenciais de Lippia sidoides e Copaifera sp. Os agentes testados apresentaram resultados similares quando comparados ao controle positivo (óleos essenciais e extratos) ou melhor efeito que o controle negativo (própolis verde). Mais estudos próximos da condição clínica e o uso de variáveis de resposta que permitam entender o mecanismo de ação são necessários, para permitir a incorporação desses agentes naturais em produtos odontológicos. O segundo capítulo teve como objetivo testar o efeito dos extratos hidroalcoólicos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade do biofilme microcosmo e na prevenção da desmineralização do esmalte. O biofilme microcosmo foi produzido em esmalte bovino, utilizando pool de saliva humana misturada à saliva de McBain (0,2% de sacarose) durante 14 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva a 100, 10 e 0,1 µg/ml e Q. grandiflora a 100 e 0,1 µg/ml reduziram a viabilidade dos microrganismos de forma semelhante ao controle positivo e significativamente maior do que o controle negativo. M. urundeuva a 1000, 100 e 0,1 µg/ml foi capaz de reduzir a contagem de Unidade formadora de colônia-UFC para Lactobacilos totais e Streptococcus mutans, enquanto a Q. grandiflora a 1000 e 1,0 µg/ml reduziu significativamente a contagem de UFC para S. mutans. Os extratos naturais não conseguiram reduzir a produção de polissacarídeos extracelulares-PEC, ácido lático e o desenvolvimento da lesão cariosa em esmalte. O terceiro capítulo teve como objetivo avaliar o efeito dos extratos hidroalcoólicos de M. urundeuva. e Q. grandiflora (sozinhos ou combinados) sobre a viabilidade do biofilme de S. mutans e na prevenção da desmineralização do esmalte. Cepa de S. mutans (ATCC 21175) foi reativada em caldo BHI. Concentração inibitória mínima, concentração bactericida mínima, concentração inibitória mínima de biofilme e concentração de erradicação mínima de biofilme foram determinadas para escolher as concentrações a serem testadas sob o modelo de biofilme. O biofilme de S. mutans (5x105 CFU/ml) foi produzido em esmalte bovino, utilizando saliva de McBain com 0,2% de sacarose durante 3 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva (isolada ou combinada) nas concentrações iguais ou superiores a 0,625 mg/ml foi capaz de reduzir a viabilidade das bactérias, enquanto que o extrato da Q. grandflora apresentou efeito antimicrobiano somente a 5 mg/ml (p<0,05). Nenhum dos extratos reduziu o desenvolvimento da lesão da cárie. Apesar dos extratos naturais terem efeito antimicrobiano, são incapazes de prevenir o desenvolvimento da lesão cariosa em esmalte.(AU)

Carbazole degradation in the soil microcosm by tropical bacterial strains

In a previous study, three bacterial strains isolated from tropical hydrocarbon-contaminated soils and phylogenetically identified as Achromobacter sp. strain SL1, Pseudomonas sp. strain SL4 and Microbacterium esteraromaticum strain SL6 displayed angular dioxygenation and mineralization of carbazole in batch cultures. In this study, the ability of these isolates to survive and enhance carbazole degradation in soil were tested in field-moist microcosms. Strain SL4 had the highest survival rate (1.8 x 107 cfu/g) after 30 days of incubation in sterilized soil, while there was a decrease in population density in native (unsterilized) soil when compared with the initial population. Gas chromatographic analysis after 30 days of incubation showed that in sterilized soil amended with carbazole (100 mg/kg), 66.96, 82.15 and 68.54% were degraded by strains SL1, SL4 and SL6, respectively, with rates of degradation of 0.093, 0.114 and 0.095 mg kg−1 h−1. The combination of the three isolates as inoculum in sterilized soil degraded 87.13% carbazole at a rate of 0.121 mg kg−1 h−1. In native soil amended with carbazole (100 mg/kg), 91.64, 87.29 and 89.13% were degraded by strains SL1, SL4 and SL6 after 30 days of incubation, with rates of degradation of 0.127, 0.121 and 0.124 mg kg−1 h−1, respectively. This study successfully established the survivability (> 106 cfu/g detected after 30 days) and carbazole-degrading ability of these bacterial strains in soil, and highlights the potential of these isolates as seed for the bioremediation of carbazole-impacted environments.

Toxicity and genotoxicity of water and sediment from streams on dotted duckweed (Landoltia punctata) / Toxicidade e genotoxicidade da água e sedimentos dos córregos em lentilha d'água (Landoltia punctata)

Most rivers are used as a source to supply entire cities; the quality of water is directly related to the quality of tributaries. Unfortunately men have neglected the importance of streams, which receive domestic and industrial effluents and transport nutrients and pesticides from rural areas. Given the complexity of the mixtures discharged into these water bodies, this study aimed to evaluate the quality of water and sediment of ten tributaries of Pirapó River, in Maringá, Paraná State, Brazil. To this end, the free-floating macrophyte Landoltia punctata (G. Meyer) Les & D.J.Crawford was used as test organism in microcosm, and the toxicity of water and sediment samples was evaluated by the relative growth rate, dry/fresh biomass ratio, and genotoxic effects (comet assay). Samples of water and sediment of each stream were arranged in microcosms with L. punctata. Seven days later, plants were collected for analysis. Nutrient levels were higher than the reference location, indicating eutrophication, but the results indicated a toxic effect for only three streams, and a genotoxic effect for all streams.

Muitos rios são utilizados como fonte de abastecimento de cidades inteiras e a qualidade de suas águas está relacionada diretamente à qualidade de seus afluentes. Infelizmente córregos têm sua importância negligenciada pelo homem sendo alvo de despejos domésticos e industriais e também do carreamento de nutrientes e pesticidas das áreas rurais. Diante da complexidade das misturas que atingem estes corpos d'água, o presente estudo teve por objetivo avaliar a qualidade da água e do sedimento de dez afluentes do rio Pirapó no município de Maringá, Estado do Paraná. Utilizou-se a macrófita aquática flutuante Landoltia punctata (G. Meyer) Les & D.J.Crawford como organismo teste em microcosmos, sendo que a toxicidade das amostras de água e sedimento foi avaliada pela taxa de crescimento relativo, pelo índice de biomassa seca por fresca, e pelos efeitos genotóxicos (ensaio de cometa). Amostras de água e sedimento de cada córrego foram dispostos em microcosmos com L. punctata. Após 7 dias as plantas foram coletadas para as análises. Os níveis de nutrientes foram maiores que o do local de referência indicando eutrofização; porém, os resultados indicaram efeito tóxico em apenas três dos córregos e genotóxico em todos os córregos.

Is nematode colonisation in the presence of Scolelepis in tropical sandy-beach sediment similar to the colonisation process in temperate sandy beaches? / A colonização dos sedimentos de praias tropicais por nematódeos na presença de Scolelepis é similar ao processo de colonização que ocorre em praias arenosas temperadas?

The role of a dominant macrobenthic polychaete, Scolelepis squamata, in the colonisation of defaunated tropical sediments by sandy-beach nematodes was investigated and compared with a previous colonisation experiment carried out on a temperate sandy beach. Experimental cylinders, equipped with lateral windows allowing infaunal colonisation, were filled with defaunated sediment containing two treatments, with and without S. squamata. These cylinders were inserted into microcosms containing sediment with indigenous meiofauna collected from the field. The treatments were incubated in the laboratory at ambient temperature and salinity for 7, 14 and 21 days. The nematode assemblages in both treatments did not differ in composition between treatments and from the natural assemblages, suggesting that all the species were equally able to colonise the experimental cores. The presence of the polychaete did not affect the development of the nematode community composition, in contrast to the results from a previous temperate-beach experiment. However, our results did not indicate whether the difference in results was caused by the different behaviour of the polychaete specimens, or by the different composition and response of the present nematode community.

Esse estudo avaliou a influência da espécie Scolelepis squamata (Polychaeta) no processo de colonização de sedimentos defaunados obtidos em uma praia arenosa tropical, além de comparar esses resultados com um estudo similar realizado em uma praia temperada. Sedimentos, previamente defaunados, foram colocados em amostradores com aberturas laterais, que permitiram a colonização da meiofauna. Foram definidos dois tratamentos, um com a presença e outro com a ausência da espécie S. squamata, além do controle. Os amostradores desses tratamentos foram alocados em unidades experimentais do tipo microcosmo, as quais continham sedimento com a meiofauna residente. Os tratamentos foram incubados em laboratório por 7, 14 e 21 dias, com condições controladas de temperatura e salinidade. Os nematódeos não apresentaram diferenças significativas em termos de composição entre os tratamentos e nem em relação ao controle, sugerindo que todas as espécies desse grupo foram, igualmente, capazes de colonizar as unidades experimentais. Esses resultados indicaram que a presença do poliqueto não afetou a estrutura da comunidade de nematódeos, o que representou um resultado contrário ao obtido para a praia temperada. No entanto, não se pode concluir se essas diferenças entre as praias estariam relacionadas ao comportamento diferencial de S. squamata ou pela presença de comunidades de nematódeos distintas nas praias.

Evaluación del tiempo de vida de bacterias potencialmente patógenas en sedimentos marinos / Evaluation of life-span of potentially pathogenic bacteria in marine sediments

El presente trabajo tuvo por objeto determinar el período de supervivencia en sedimentos marinos de bacterias con riesgo biológico o indicadoras. Se tomaron muestras de sedimento marino intermareal en dos playas de la ciudad de Comodoro Rivadavia, Argentina; una sometida a contaminación por efluentes domiciliarios, y otra sin pruebas de contaminación evidente. Sobre éstos se realizó una caracterización inicial por recuento de bacterias en diferentes medios de cultivo (TSA, VRB, VRBG, TCBS, agar bilis esculina y agar cetrimide) y por su identificación. También se realizó un estudio de supervivencia de los microorganismos de interés en microcosmos. Estos fueron monitoreados por conteo de bacterias y por identificación de los integrantes de interés. Los principales resultados indicaron que existe una marcada diferencia en la composición de la comunidad bacteriana entre ambas muestras. Los géneros bacterianos en la muestra contaminada fueron Aeromonas, Salmonella, Pseudomonas, Escherichia, Ochrobactum y Klebsiella, y en la playa no contaminada Bacillus, Paenibacillus, Micrococcus, Photobacterium y Vibrio. El ensayo en microcosmos demostró que, luego de 65 días, la comunidad bacteriana de la playa contaminada fue similar a la del sedimento sin contaminación, indicando que al retirar la fuente de contaminación, es probable la recuperación de la zona afectada.

The present study had the purpose of determining the survival period of bacteria which represent a biological risk and are indicators of contamination. Inter-tide marine sediment samples were taken at two beaches of the Comodoro Rivadavia city in Argentine; one the beaches was submitted to contamination due to household effluents, and the other without proof of evident contamination. These samples were initially characterized by bacterial counts in different culture media (TSA, VRB, TCBS, bile esculin agar and cetrimide agar) and by their identification. A study of the survival of the target bacteria in microcosm was also done, monitored by bacterial counts and identification of these bacteria. The main results indicated that there was a marked difference in the composition of the bacterial community between the two samples. In the contaminated sample the bacterial genus were Aeromonas, Salmonella, Pseudomonas, Escherichia, Ochrobactum and Klebsiella, and in the non-contaminated beach Bacillus, Paenibacillus, Micrococcus, Photobacterium and Vibrio. The microcosm assay showed that after 65 days the bacterial community of the contaminated beach was similar to that of the non-contaminated beach sediment, indicating that by withdrawing the contamination source, the affected zone can probably be recovered.

Effects of Mycorrhizal and Endophytic Fungi on Plant Community: a Microcosm Study

This study was conducted to investigate the effects of foliar endophytic fungi and arbuscular mycorrhizal fungi (AMF) on plant community structure in experimental microcosms containing an assemblage of five species of plants (Oenothera odorata, Plantago asiatica, Trifolium repens, Isodon japonicas and Aster yomena). Leaves of Sasa borealis, Potentilla fragarioides, and Viola mandshurica were collected in Chungbuk, Korea. Endophytic fungi were isolated from the surface sterilized leaves and identified to species level using molecular and morphological techniques. Four isolates of the endophytic fungi were inoculated to the leaves of host plants in the microcosms. Also, three species of AMF spores were extracted from pure cultures and the mixture of the three species inoculated to the roots of the plants. After four months of growth in a green house, effects of both symbiotic fungi on plant species diversity, community composition and productivity were examined. The plant species diversity showed significant differences with inoculation of the symbiotic fungi. Results indicate that AMF significantly affect plant productivity and plant community structure.