Density and diversity of filamentous fungi in the water and sediment of Araçá bay in São Sebastião, São Paulo, Brazil / Densidade e diversidade de fungos filamentosos na água e sedimento da Baía do Araçá, São Sebastião (SP), Brasil

Abstract Araçá Bay, located in the city of São Sebastião, São Paulo, Brazil, is a protected area of substantial complexity. It represents the last remaining mangrove swamp preserve between the cities of Bertioga and Ubatuba on the northern coast of São Paulo State. This mangrove swamp has specific physical and chemical properties, and it shelters a wide variety of life, including fungi. These microorganisms are present in a variety of species with different morphophysiological features, and they have the ability to produce enzymes of biotechnological importance. The goal of this study was to quantify, isolate, and identify filamentous fungi in water and sediment samples from the Araçá Bay mangrove swamp in São Sebastião. Two samplings were performed in the summer and two were performed in the winter. The samples were collected from intertidal zones, and dissolved oxygen (DO), temperature, salinity, and pH were measured in situ. The spread plate technique was used to inoculate the samples collected on plates with a potato dextrose agar (PDA) medium. A total of 208 colonies (68 from water samples and 140 from sediment samples) were isolated, and they were identified based on their morphological characteristics. Filamentous fungus density was higher in the sediment than in the water, and the samplings performed in the winter revealed a higher density than those performed in the summer. Though some of the environmental parameters were not ideal for fungal development, a high quantity of growth was nevertheless observed. When the isolated colonies were analyzed, the greatest diversity and species richness were found in the summer samples. The genera identified in all of the samples were Aspergillus, Penicillium, Cladosporium, and Fusarium. The pathogenic species found from these genera were Aspergillus fumigatus, A. terreus, Penicillium citrinum, and P. chrysogenum. These species are also able to produce enzymes that offer a variety of applications. The fungal community described herein represents the diversity found in this mangrove swamp during the period studied. Many of the fungus species found are pathogenic and may be useful due to their ability to produce specific enzymes applicable in the biotechnological and pharmaceutical industries.

Resumo A Baía do Araçá, localizada no município de São Sebastião (SP), é uma região protegida, de grande complexidade, mantendo o último remanescente preservado de manguezal entre Bertioga e Ubatuba, litoral Norte do Estado de São Paulo. Manguezal caracteriza-se com propriedades físico-químicas específicas e abriga grande diversidade de seres vivos, entre eles os fungos. Estes microrganismos apresentam grande variedade de espécies e diferentes características morfofisiológicas com capacidade de produzir algumas enzimas de importância biotecnológica. Este trabalho teve como objetivo quantificar, isolar e identificar fungos filamentosos em amostras de água e de sedimento do manguezal da Baía do Araçá, São Sebastião, (SP). Foram realizadas 2 coletas no verão e 2 no inverno na região entremarés, medidos "in situ" os parâmetros oxigênio dissolvido (OD), temperatura, salinidade e pH. Utilizou-se a Técnica "Spread Plate" para inocular os materiais coletados, em placas com meio Potato Dextrose Agar (PDA). Foram isoladas 208 colônias (68 das amostras de água e 140 das amostras de sedimento) e identificadas pelas características morfofisiológicas. As densidades de fungos filamentosos foram maiores no sedimento em relação à água e nas coletas realizadas no inverno demonstraram maiores densidades do que no verão. Apesar de alguns parâmetros ambientais não se apresentarem ideais para o desenvolvimento dos fungos, observou-se alta quantidade de crescimento nas amostragens. Analisando as colônias isoladas, a maior diversidade e riqueza foram observadas na amostragem coletado no verão. Foram observados os gêneros Aspergillus sp, Penicillium sp, Cladosporium sp e Fusarium sp em todas as amostragens, dos quais as espécies Aspergillus fumigatus, A. terreus, Penicillium citrinum e P. chrysogenum identificados neste trabalho são considerados patogênicos mas também são espécies capazes de produzir enzimas aplicáveis em diversas atividades. A comunidade fúngica descrita apresenta a diversidade encontrada neste manguezal em relação à variedade ambiental no período estudado, sendo que muitos apresentam patogenicidade e podem ser uteis pela sua capacidade de produzir enzimas específicas aplicáveis nos estudos biotecnológicos e farmacêuticos.

Comparison of bacterial densities and resistance in different beach compartments: should water be our main concern? / Comparação de densidade e resistência bacterianas em diferentes compartimentos de praia: a água deve ser nossa principal preocupação?

Coastal regions are very important, since they provide food, enable economic and leisure activities however, the increase in urbanization of coastal areas are accompanied by great volumes of organic effluent, which is sometimes discharged in natura in water bodies, increasing the risk of the presence of pathogenic resistant bacteria in marine environments. In fact, recent studies showed higher bacterial densities in sediments than in water, since its present more favorable conditions for bacterial survivor (e.g. sun protection and predation). In addition, bivalves tend to accumulate suspended bacteria from the water, since they are filter feeding organisms. Thus, the present study evaluated densities and resistance to antibiotics of Enterococcus sp., Escherichia coli and Aeromonas sp. in water, sediment and mussels samples. Samples were collected at Praia dos Sonhos (Itanhaém) and Ubuqueçaba Island (Santos). Bacterial densities were determined by Membrane Filter Technique and the isolated strains were submitted to antibiotic sensibility test. Bacterial densities were lower in water and higher in sediments and mussels samples. Bacterial strains from Santos presented higher frequencies of resistance than those isolated from Itanhaém (less impacted area). Aeromonas strains were more resistant to Cefalotin and Cefuroxin, Enterococcus to Gentamicin and Streptomicin, and E. coli to Vancomicin and Eritromicin. The results obtained point to the need to establish public policies, laws and monitoring programs about the microbiological quality of mollusks and sediments, including the use of Enterococcus sp as microbiological indicator as well as about the resistance of the bacteria present in these environments

As regiões costeiras são muito importantes, uma vez que proporcionam alimentos, permitem atividades econômicas e de lazer, no entanto, o aumento da urbanização das áreas costeiras é acompanhado por grandes volumes de efluentes orgânicos, que às vezes são descarregados in natura em corpos d'água, aumentando o risco da presença de bactérias patogênicas e resistentes em ambientes marinhos. De fato, estudos recentes mostraram maiores densidades bacterianas nos sedimentos do que na água, pois apresentam condições mais favoráveis para a sobrevivência bacteriana (e.g. proteção solar e de predação). Além disso, os bivalves tendem a acumular bactérias suspensas da água, pois são organismos que alimentam-se por filtração. Assim, o presente estudo avaliou densidades e resistência a antibióticos de Enterococcus sp., Escherichia coli e Aeromonas sp. em amostras de água, sedimentos e mexilhões. As amostras foram coletadas na Praia dos Sonhos (Itanhaém) e na Ilha Ubuqueçaba (Santos). As densidades bacterianas foram determinadas pela técnica de filtro de membrana e as cepas isoladas foram submetidas ao teste de sensibilidade aos antibióticos. As densidades bacterianas foram menores na água e maiores nas amostras de sedimentos e mexilhões. As cepas bacterianas de Santos apresentaram maiores frequências de resistência do que as isoladas de Itanhaém (área menos impactada). As cepas de Aeromonas foram mais reistentes à Cefalotina e Cefuroxime, Enterococcus à Gentamicina e Estreptomicina, e E. coli à Vancomicina e Eritromicina. Os resultados obtidos apontam para a necessidade de estabelecer políticas públicas, leis e programas de monitoramento relativos à qualidade microbiológica de moluscos e sedimentos, incluindo o uso de Enterococcus sp. como indicador microbiológico, bem como sobre a resistência das bactérias presentes nesses ambientes

Diversity and Distribution of Heavy Metal-Resistant Bacteria in Polluted Sediments of the Araça Bay, São Sebastião (SP), and the Relationship Between Heavy Metals and Organic Matter Concentrations.

Heavy metals influence the population size, diversity, and metabolic activity of bacteria. In turn, bacteria can develop heavy metal resistance mechanisms, and this can be used in bioremediation of contaminated areas. The purpose of the present study was to understand how heavy metals concentration influence on diversity and distribution of heavy metal-resistant bacteria in Araça Bay, São Sebastião, on the São Paulo coast of Brazil. The hypothesis is that activities that contribute for heavy metal disposal and the increase of metals concentrations in environment can influence in density, diversity, and distribution of heavy metal-resistant bacteria. Only 12 % of the isolated bacteria were sensitive to all of the metals tested. We observed that the highest percentage of resistant strains were in areas closest to the São Sebastião channel, where port activity occurs and have bigger heavy metals concentrations. Bacterial isolated were most resistant to Cr, followed by Zn, Cd, and Cu. Few strains resisted to Cd levels greater than 200 mg L(-1). In respect to Cr, 36 % of the strains were able to grow in the presence of as much as 3200 mg L(-1). Few strains were able to grow at concentrations of Zn and Cu as high as 1600 mg L(-1), and none grew at the highest concentration of 3200 mg L(-1). Bacillus sp. was most frequently isolated and may be the dominant genus in heavy metal-polluted areas. Staphylococcus sp., Planococcus maritimus, and Vibrio aginolyticus were also isolated, suggesting their potential in bioremediation of contaminated sites.

Densities and antimicrobial resistance of Escherichia coli isolated from marine waters and beach sands.

Bacterial resistance is a rising problem all over the world. Many studies have showed that beach sands can contain higher concentration of microorganisms and represent a risk to public health. This paper aims to evaluate the densities and resistance to antimicrobials of Escherichia coli strains, isolated from seawater and samples. The hypothesis is that microorganisms show higher densities in contaminated beach sands and more antimicrobial resistance than the water column. Density, distribution, and antimicrobial resistance of bacteria E. coli were evaluate in seawater and sands from two recreational beaches with different levels of pollution. At the beach with higher degree of pollution (Gonzaguinha), water samples presented the highest densities of E. coli; however, higher frequency of resistant strains was observe in wet sand (71.9 %). Resistance to a larger number of antimicrobial groups was observe in water (betalactamics, aminoglycosides, macrolides, rifampicins, and tetracyclines) and sand (betagalactamics and aminoglycosids). In water samples, highest frequencies of resistance were obtain against ampicilin (22.5 %), streptomycin (15.0 %), and rifampicin (15.0 %), while in sand, the highest frequencies were observe in relation to ampicilin (36.25 %) and streptomycin (23.52 %). At the less polluted beach, Ilha Porchat, highest densities of E. coli and higher frequency of resistance were obtain in wet and dry sand (53.7 and 53.8 %, respectively) compared to water (50 %). Antimicrobial resistance in strains isolated from water and sand only occurred against betalactamics (ampicilin and amoxicilin plus clavulanic acid). The frequency and variability of bacterial resistance to antimicrobials in marine recreational waters and sands were related to the degree of fecal contamination in this environment. These results show that water and sands from beaches with a high index of fecal contamination of human origin may be potential sources of contamination by pathogens and contribute to the dissemination of bacterial resistance.

Heavy metals and TPH effects on microbial abundance and diversity in two estuarine areas of the southern-central coast of São Paulo State, Brazil.

Coastal areas may be impacted by human and industrial activities, including contamination by wastewater, heavy metals and hydrocarbons. This study aimed to evaluate the impact of hydrocarbons (TPH) and metals on the microbiota composition and abundance in two estuarine systems in the coast of São Paulo: the Santos (SE) and Itanhaém (IE) estuaries. The SE was found to be chronically contaminated by heavy metals and highly contaminated by hydrocarbons. This finding was correlated with the increased density of cyanobacteria in sediments and suggests the possible use of cyanobacteria for bioremediation. These contaminants influence the density and composition of estuarine microbiota that respond to stress caused by human activity. The results are troubling because quantitative and qualitative changes in the microbiota of estuarine sediments may alter microbiological processes such as decomposition of organic matter. Moreover, this pollution can result in damage to the environment, biota and human health.

Densidade de Enterococcus sp em águas recreacionais e areias de praias do município de São Vicente-SP, Brasil e sua relação com parâmetros abióticos / Density of Enterococcus sp in recreational waters and beach sandin São Vicente-SP, Brazil and its relationship to abiotic parameters

As cidades litorâneas atraem inúmeros turistas pela sua beleza cênica e atrativos recreacionais, incrementando a fonte de renda dos municípios. Dessa forma, é evidente a necessidade de programas de monitoramento da qualidade de suas praias, a fim de assegurara saúde dos banhistas. Apesar de haver um monitoramento eficaz para as águas recreacionais marinhas, ainda não existem, no Brasil, programas que atestem a qualidade das areias das praias. Nesse sentido, o presente estudo objetivou analisar a densidade de bactérias do gênero Enterococcus nas areias e águas de duas praias do município de São Vicente-SP, Brasil correlacionando com parâmetros abióticos (temperatura, salinidade, granulometria, matéria orgânica e variação de maré). Amostras de água e areia foram coletadas durante o mês de fevereiro de 2006 nas praias do Gonzaguinha e da Ilha Porchat, e as densidades das bactérias,determinadas com base na técnica de membrana filtrante. Temperatura e salinidade foram medidas in situ com o auxílio de um termômetro e um refratômetro, enquanto que granulometria e matéria orgânica foram determinadas em laboratório, de acordo com os métodos descritos por Suguio e Dean. Houve diferenças significativas entre as densidades encontradas na água e nas areias(p=0.004), sendo aproximadamente 20 vezes superior na areia das praias do Gonzaguinha. Resultado semelhante foi obtido para as amostras das praias da Ilha Porchat, sendo as densidades encontradas nas areias cerca de três vezes superior às encontradas nas amostras de água. Em ambas as praias, salinidade e temperatura correlacionaram-se de maneira negativa com as densidades obtidas, o que sugere que há um efeito deletério desses parâmetros sobre a população bacteriana. O presente estudo demonstra que há uma grande necessidade de estabelecimento de programas de monitoramento para areias de praias, visto que atualmente a avaliação da qualidade das praias é parte importante de programas de gerenciamento costeiro integrado.

Coastal cities attract a large number of tourists for their beautiful landscape and recreational activities, increasing the municipalities’income source. Thus, a need is clear for the establishment of beach quality monitoring programs to ensure bathers health. Although there is an effective monitoring program for recreational waters in Brazil there are no programs to certify the quality of beach sands. In this sense, the aim of this work was to analyze the density of bacteria from the genus Enterococcus in both sand and water from two beaches from São Vicente, São Paulo (Brazil) and correlate these densities to abiotic parameters such as: temperature, salinity, particle size, organic matter and tides). Water and sand samples were collected during February 2006 on the beaches of Gonzaguinha e Ilha Porchat and bacterial densities were determined by membrane filter technique. Temperature and salinity were measured in situwith a thermometer and a refractometer while particle size and organic matter were determined according to methods described by Suguio and Dean. There were significant differences between densities found in water and sand (p=0.004), being approximately 20times higher in Gonzaguinha’s beach sands. Similar results were found for Ilha Porchat beach samples, being the densities found insand 3 times greater than those found in water. Both beaches showed a negative correlation between bacterial densities and salinity and temperature, suggesting a deleterious effect of these parameters on the bacterial community. On the other hand, no correlation was found between organic matter and particle size and bacterial densities. This work demonstrates that there is a great necessity for monitoring beach sand since the evaluation of beach quality is an important part of integrated coastal management programs.

Diversidade de microrganismos indicadores utilizados na avaliaçãoda contaminação fecal de areias de praias recreacionais marinhas:estado atual do conhecimento e perspectivas / Diversity of indicator microorganisms in the evaluation of sea recreational beach sand fecalcontamination: current knowledge and perspectives / Diversidad de los microorganismos indicadores utilizados en la evaluación de la contaminación fecal de las arenas de playas marinas recreacionales: conocimiento actual y perspectivas

Embora a qualidade de águas recreacionais marinhas já seja monitorada por programas implantados em vários estados brasileiros,incluindo o Estado de São Paulo, pouca atenção tem sido dada às areias de praias, as quais têm sido desconsideradas do ponto de vista da saúde pública. Entretanto, esse panorama vem mudando nos últimos anos devido ao aumento dos casos de micoses e infecções bacterianas contraídas por pessoas que frequentam as praias e utilizam suas areias como local de recreação. Isso tem ocasionado maiores preocupações com a contaminação desse ambiente, também mensurável pelo aumento do número de trabalhos científicos sobre a microbiota de sedimentos e areias depraias recreacionais. Atualmente, sabe-se que, de maneira geral, esses sedimentos contêm mais microrganismos do que a própria coluna de água, sendo, portanto, fontes potenciais de contaminação de humanos por microrganismos patogênicos. Os resultados de trabalhos realizados emvários países são preocupantes e têm demonstrado a necessidade da determinação de padrões e limites para que sejam implantados programasde monitoramento da qualidade microbiológica das areias de praias. Tal preocupação é especialmente destacada no Brasil, país de clima tropical onde milhares de praias, utilizadas para recreação, se estendem por quase oito mil quilômetros de litoral. No contexto da Baixada Santista,estudos realizados têm mostrado que, em determinadas situações, as areias de praias podem conter mais microrganismos de que suas águas, opode oferecer riscos à saúde de seus usuários.

Although the quality of sea recreational waters is already monitored by programs implanted in some Brazilian states, including the State São Paulo, little attention has been given to beach sands, which have been disregarded from the point of view of public health. However, this panorama is changing in recent years due to an increasing number of cases of mycoses and bacterial infections affecting people who frequent beaches and use sands as recreation places. This has caused greater concerns with the contamination of this environment, also measurable by the increase of the number of scientific works on sediments and recreational beach sands microbiota. Currently one knows that in general these sediments contain more microorganisms than the water and are therefore potential sources of contamination of human beings by pathogenic microorganisms. The results of works carried through in some countries are worrying, and have demonstrated the necessity of establishing standards and limits so that monitoring programs of the microbiological quality of beach sands are implanted. Such concern is especially high in Brazil, a country of a tropical climate where thousands of beaches, used for recreation, extend for almost eight thousand kilometers of the coast. In the context of Baixada Santista, studies carried through have shown that in certain situations beach sands can contain more microorganismsthan waters and may be a risk to the health of users.

Aunque la calidad de las aguas marinas recreacionales sea supervisada ya por programas implantados en algunos estados brasileños, incluyendo el estado São Paulo, poca atención se ha dado a las arenas de las playas, que se han desatendido desde el punto de vista de la salud pública. Sin embargo, este panorama está cambiando estos últimos años debido a un número cada vez mayor de casos de micosis y infecciones bacterianas que afectan a la gente que usa las playas y las arenas como lugares de recreo. Esto ha causado mayores preocupaciones con la contaminación de este ambiente, también mensurable por el aumento del número de trabajos científicos sobre los microorganismos de sedimentosy de las arenas de playas recreacionales. Se sabe hoy que en general estos sedimentos contienen más microorganismos que el agua y que son por lo tanto fuentes potenciales de contaminación de seres humanos por microorganismos patógenos. Los resultados de los trabajos ejecutados en algunos países son alarmantes, y han demostrado la necesidad de establecer estándares y límites para implantar programas de supervisión de la calidad microbiológica de las arenas de las playas. Tal preocupación es especialmente alta en el Brasil, un país de clima tropical en donde millares de playas, usados para el recreo, se extienden en casi ocho mil kilómetros de la costa. En el contexto de Baixada Santista, los estudiosejecutados han demostrado que en ciertas situaciones las arenas de la playa pueden contener más microorganismos que las aguas y que pueden ser un riesgo a la salud de usuarios.

Antimicrobial resistance of heterotrophic marine bacteria isolated from seawater and sands of recreational beaches with different organic pollution levels in southeastern Brazil: evidences of resistance dissemination.

Antimicrobial resistance of marine heterotrophic bacteria to different antimicrobials agents were evaluated in seawater, dry and wet sands from three marine recreational beaches with different pollution levels. In all studied beaches, the greatest frequencies of resistance were found in relation to penicillin. On Gonzaguinha, the most polluted beach, 72.3% of all isolated strains showed simple resistance, whilst 8.33% had multiple resistance. The values found on Ilha Porchat beach, were 70.8% and 6.9% for simple and multiple resistances, respectively. On Guaraú, the less polluted beach, only 35.3% of isolated strains had simple resistance. Multiple resistance was not observed. While samples from Gonzaguinha and Ilha Porchat beach showed isolated strains resistant to seven and six different antimicrobial agents, respectively, samples from Guaraú beach were resistant only to penicillin and erytromicin. The positive correlations obtained between the degree of seawater contamination and frequency and variability of bacterial resistance indicate that polluted marine recreational waters and sands are sources of resistant bacteria contributing thus, to the dissemination of bacterial resistance.