Cryptic speciation in populations of the genus Aphyocharax (Characiformes: Characidae) from eastern Amazon coastal river drainages and surroundings revealed by single locus species delimitation methods

Recent studies in eastern Amazon coastal drainages and their surroundings have revealed new fish species that sometimes exhibit little morphological differentiation (cryptic species). Thus, we used a DNA-based species delimitation approach to test if populations showing the morphotype and typical character states of the Aphyocharax avary holotype correspond either to A. avary or A. brevicaudatus, two known species from the region, or if they form independent lineages, indicating cryptic speciation. WP and GMYC analyses recovered five lineages (species) in the ingroup, while a bPTP analysis delimited three lineages. ABGD analyses produced two possible results: one corroborating the WP and GMYC methods and another corroborating the bPTP method. All methods indicate undescribed cryptic species in the region and show variation from at least 1 to 4 species in the ingroup, depending on the approach, corroborating previous studies, and revealing this region as a possible hotspot for discovering undescribed fish species.(AU)

Estudos recentes nas drenagens costeiras da Amazônia oriental e seus arredores revelaram novas espécies de peixes que às vezes exibem pouca diferenciação morfológica (espécies crípticas). Assim, usamos uma abordagem de delimitação de espécies baseada em DNA para testar se as populações que apresentam o morfotipo e os estados de caráter típicos do holótipo Aphyocharax avary correspondem a A. avary ou A. brevicaudatus, duas espécies conhecidas da região, ou se formam linhagens independentes, indicando especiação críptica. As análises de WP e GMYC recuperaram cinco linhagens (espécies) no grupo interno, enquanto uma análise de bPTP delimitou três linhagens. As análises ABGD produziram dois resultados possíveis: um corroborando os métodos WP e GMYC e outro corroborando o método bPTP. Todos os métodos indicam espécies crípticas não descritas na região e apresentam variação de pelo menos uma a quatro espécies no grupo interno, dependendo da abordagem, corroborando estudos anteriores, e revelando esta região como um possível "hotspot" para descoberta de espécies de peixes não descritas.(AU)

Detection of multidrug-resistant Enterobacteriaceae isolated from river waters flowing to the Guanabara Bay and from clinical samples of hospitals in Rio de Janeiro, Brazil / Detección de enterobacterias multirresistentes aisladas en aguas de los ríos que desembocan en la bahía de Guanabara y en muestras de hospitales de Río de Janeiro, Brasil

Abstract Introduction: The use of antibiotics in humans, animal husbandry and veterinary activities induces selective pressure leading to the colonization and infection by resistant strains. Objective: We evaluated water samples collected from rivers of the Guanabara Bay, which have suffered minor and major environmental degradation, and clinical samples of hospital origin to detect evidence of the presence of resistance genes to aminoglycosides, beta-lactam antibiotics and fluoroquinolones in strains of Klebsiella pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae and Escherichia coli. Materials and methods: For isolation of the water strains we employed culture media containing 32 μg/ml cephalotin and 8 μg/ml gentamicin. The strains from clinical materials were selected using culture media containing 8 μg/ml gentamicin. The strains were identified and subjected to antimicrobial susceptibility testing (AST), plasmid DNA extraction and polymerase chain reaction (PCR) to detect genes encoding enzymes modifying aminoglycosides (EMA), extended-spectrum beta-lactamases (ESBL) and plasmid mechanisms of quinolone resistance (PMQR). Results: The AST of the isolates recovered from water samples showed multidrug-resistance profiles similar to those found in isolates recovered from clinical materials. All isolates from water samples and 90% of the isolates from clinical samples showed at least one plasmid band. In the PCR assays, 7.4% of the isolates recovered from water samples and 20% of those from clinical materials showed amplification products for the three antimicrobial classes. Conclusion: We believe that the detection of microorganisms presenting genetic elements in environments such as water is necessary for the prevention and control of their dissemination with potential to infect humans and other animals in eventual contact with these environments.

Resumen Introducción. El uso de antibióticos en seres humanos, en la industria pecuaria y en las actividades veterinarias induce una presión selectiva que resulta en la colonización e infección con cepas resistentes. Objetivo. Determinar la presencia de genes de resistencia a aminoglucósidos, betalactámicos y fluoroquinolonas en cepas de Klebsiella pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae y Escherichia coli, obtenidas de muestras de agua de los ríos que desembocan en la bahía de Guanabara y de muestras clínicas de hospitales de Río de Janeiro. Materiales y métodos. En la selección de las cepas resistentes obtenidas de las muestras de agua de los ríos, se emplearon medios de cultivo que contenían 32 μg/ml de cefalotina y 8 μg/ ml de gentamicina. En el caso de las muestras de especímenes clínicos, se usaron medios de cultivo que contenían 8 μg/ml de gentamicina. Las cepas se identificaron y se sometieron a pruebas de sensibilidad antimicrobiana, extracción de ADN plasmídico y pruebas de reacción en cadena de la polimerasa (PCR) para detectar los genes que codifican aquellas enzimas que modifican los aminoglucósidos, las betalactamasas de espectro extendido (BLEE) y los mecanismos de resistencia a las quinolonas mediados por plásmidos. Resultados. Se encontraron perfiles de resistencia a los antimicrobianos similares en los dos grupos. En todas las bacterias obtenidas de las muestras de agua y en 90 % de las muestras clínicas, se evidenciaron bandas de plásmidos asociados con la transferencia de genes de resistencia. En las pruebas de PCR, se obtuvieron productos de amplificación de los genes de resistencia para las tres clases de antimicrobianos analizados, en el 7,4 % de las bacterias recuperadas de las muestras de agua y en el 20 % de aquellas recuperadas de las muestras clínicas. Conclusión. La detección de microorganismos con elementos genéticos que confieren resistencia a los antibióticos en ambientes como el agua, es una estrategia necesaria para prevenir y controlar la diseminación de estos agentes patógenos con potencial para infectar a humanos y a otros animales en dichos ambientes.

Study of pandrug and heavy metal resistance among E. coli from anthropogenically influenced Delhi stretch of river Yamuna

Abstract Escalating burden of antibiotic resistance that has reached new heights present a grave concern to mankind. As the problem is no longer confined to clinics, we hereby report identification of a pandrug resistant Escherichia coli isolate from heavily polluted Delhi stretch of river Yamuna, India. E. coli MRC11 was found sensitive only to tobramycin against 21 antibiotics tested, with minimum inhibitory concentration values >256 µg/mL for amoxicillin, carbenicillin, aztreonam, ceftazidime and cefotaxime. Addition of certain heavy metals at higher concentrations were ineffective in increasing susceptibility of E. coli MRC11 to antibiotics. Withstanding sub-optimal concentration of cefotaxime (10 µg/mL) and mercuric chloride (2 µg/mL), and also resistance to their combinatorial use, indicates better adaptability in heavily polluted environment through clustering and expression of resistance genes. Interestingly, E. coli MRC11 harbours two different variants of blaTEM (blaTEM-116 and blaTEM-1 with and without extended-spectrum activity, respectively), in addition to mer operon (merB, merP and merT) genes. Studies employing conjugation, confirmed localization of blaTEM-116, merP and merT genes on the conjugative plasmid. Understanding potentialities of such isolates will help in determining risk factors attributing pandrug resistance and strengthening strategic development of new and effective antimicrobial agents.

Escherichia coli is not a suitable fecal indicator to assess water fecal contamination by otters / Escherichia coli não é um indicador adequado para avaliar a contaminação fecal da água por lontras

Abstract The detection of pathogenic microorganisms in aquatic environments is extremely relevant in terms of public health. As these laboratorial methodologies are usually difficult, expensive and time-consuming, they are frequently replaced by the assessment of fecal indicator bacteria, such as Escherichia coli. This study aimed to assess the presence of E. coli in fecal samples from Neotropical otters, to evaluate its potential as fecal indicator to be applied to the determination of water microbiological quality in areas where otters' populations are high. Twenty-six otter fecal samples, collected in Alto Paranapanema river basin, São Paulo State, Brazil, were analyzed for the presence of E. coli, using conventional bacteriological methods. Only 8 scat samples (30%) were E. coli positive, indicating that this microorganism is not a suitable fecal indicator to assess water fecal contamination by Neotropical otters, and should not be used to infer the presence of otter related pathogens in waters.

Resumo A detecção de microrganismos patogênicos em ambientes aquáticos é extremamente importante em termos de saúde pública. Como estas metodologias laboratoriais são geralmente difíceis de realizar, dispendiosas e demoradas, são frequentemente substituídas pela avaliação de bactérias indicadoras de contaminação fecal, tais como Escherichia coli. Este estudo teve como objetivo avaliar a presença de E. coli em amostras fecais de lontras Neotropicais, para avaliar o seu potencial como indicador fecal e poder ser aplicado para a determinação da qualidade microbiológica da água em áreas onde as populações de lontras são numerosas. Vinte e seis amostras de fezes de lontra, coletadas na bacia do Alto Paranapanema, Estado de São Paulo, Brasil, foram analisadas quanto à presença de E. coli, utilizando métodos bacteriológicos convencionais. Apenas oito amostras de fezes (30%) se revelaram positivas para E. coli, indicando que este microrganismo não é um indicador fecal adequado para avaliar a contaminação fecal da água por lontras Neotropicais, e não deve ser usada para inferir a presença de agentes patogênicos relacionados com lontra em águas.

Bioremediation of polycyclic aromatic hydrocarbon (PAH) compounds: (acenaphthene and fluorene) in water using indigenous bacterial species isolated from the Diep and Plankenburg rivers, Western Cape, South Africa

Abstract This study was conducted to investigate the occurrence of PAH degrading microorganisms in two river systems in the Western Cape, South Africa and their ability to degrade two PAH compounds: acenaphthene and fluorene. A total of 19 bacterial isolates were obtained from the Diep and Plankenburg rivers among which four were identified as acenaphthene and fluorene degrading isolates. In simulated batch scale experiments, the optimum temperature for efficient degradation of both compounds was determined in a shaking incubator after 14 days, testing at 25 °C, 30 °C, 35 °C, 37 °C, 38 °C, 40 °C and 45 °C followed by experiments in a Stirred Tank Bioreactor using optimum temperature profiles from the batch experiment results. All experiments were run without the addition of supplements, bulking agents, biosurfactants or any other form of biostimulants. Results showed that Raoultella ornithinolytica, Serratia marcescens, Bacillus megaterium and Aeromonas hydrophila efficiently degraded both compounds at 37 °C, 37 °C, 30 °C and 35 °C respectively. The degradation of fluorene was more efficient and rapid compared to that of acenaphthene and degradation at Stirred Tank Bioreactor scale was more efficient for all treatments. Raoultella ornithinolytica, Serratia marcescens, Bacillus megaterium and Aeromonas hydrophila degraded a mean total of 98.60%, 95.70%, 90.20% and 99.90% acenaphthene, respectively and 99.90%, 97.90%, 98.40% and 99.50% fluorene, respectively. The PAH degrading microorganisms isolated during this study significantly reduced the concentrations of acenaphthene and fluorene and may be used on a larger, commercial scale to bioremediate PAH contaminated river systems.

Pathogenic Vibrio species isolated from estuarine environments (Ceará, Brazil) - antimicrobial resistance and virulence potential profiles

ABSTRACT Detection of virulent strains associated with aquatic environment is a current concern for the management and control of human and animal health. Thus, Vibrio diversity was investigated in four estuaries from state of Ceará (Pacoti, Choró, Pirangi and Jaguaribe) followed by antimicrobial susceptibility to different antimicrobials used in aquaculture and detection of main virulence factors to human health. Isolation and identification were performed on TCBS agar (selective medium) and dichotomous key based on biochemical characteristics, respectively. Nineteen strains of genus Vibrio were catalogued. Vibrio parahaemolyticus (Choró River) and V. alginolyticus (Pacoti River) were the most abundant species in the four estuaries. All strains were submitted to disk diffusion technique (15 antimicrobials were tested). Resistance was found to: penicillin (82%), ampicillin (54%), cephalotin (7%), aztreonan (1%), gentamicin, cefotaxime and ceftriaxone (0.5%). Five pathogenic strains were chosen to verification of virulence factors. Four estuaries showed a high abundance of species. High number of tested positive strains for virulence is concerning, since some of those strains are associated to human diseases, while others are known pathogens of aquatic organisms.

Avaliação da qualidade da água do Rio dos Sinos (Brasil) por meio do teste de micronúcleos em Cyprinus carpio e de análises físico-químicas e microbiológicas / Evaluation of water quality of the Sinos River (Brazil) by micronucleus test in Cyprinus carpio and physicochemical and microbiological analysis

O Rio dos Sinos está localizado na região sul do Brasil, possui aproximadamente 190 km de extensão e fornece água para atividades agrícolas, industriais e para o consumo de mais de um milhão de habitantes. Este rio é considerado um dos mais poluídos do Brasil e, portanto, estudos para avaliar a qualidade da água ao longo de sua extensão são necessários. O teste de micronúcleos em eritrócitos de peixes tem sido utilizado com sucesso para detectar a presença de poluentes mutagênicos nos ambientes aquáticos. O objetivo do presente estudo foi avaliar a qualidade da água do Rio dos Sinos por meio do teste de micronúcleos em Cyprinus carpio bem como de análises físico-químicas e microbiológicas. Foram coletadas amostras de água nos trechos superior (Caraá), médio (Parobé) e inferior (Novo Hamburgo) do Rio dos Sinos em dezembro de 2013. As amostras foram transportadas para o laboratório para análise de 14 parâmetros de qualidade da água bem como para a exposição de espécimes de C. carpio por 72h em aquários. Não foram verificadas diferenças significativas na frequência de micronúcleos e anormalidades nucleares nos grupos expostos a água do rio em comparação ao grupo controle. Apenas um parâmetro de qualidade da água foi observado em desacordo com a legislação no trecho superior do rio, enquanto que nos trechos médio e inferior foram seis e cinco parâmetros, respectivamente. Os resultados demonstram que o Rio dos Sinos apresenta redução da qualidade da água ao longo de sua extensão e indicam ausência de potencial genotóxico no período amostrado.

The Sinos River is located in South of Brazil, it has about 190 km of extension and provides water for agricultural and industrial activities, and for consumption of more than one million inhabitants. This river is considered one of the most polluted rivers in Brazil; therefore, studies aiming to evaluate the water quality along its course are necessary. The micronucleus test in fish erythrocytes has been successful used to detect mutagenic pollutants in the aquatic environment. The objective of this study was to evaluate the water quality of the Sinos River by the micronucleus test in Cyprinus carpio, as well as physicochemical and microbiological analyses. Water samples were collected in the upper (Caraá municipality), middle (Parobé municipality) and lower (Novo Hamburgo municipality) sections of the Sinos River, in December 2013. The samples were transported to the laboratory for analysis of 14 water quality parameters and for exposure of C. carpio for 72 hours in aquaria. Significant differences in micronucleus and nuclear abnormalities frequencies were not found between the control and the exposed groups. Only one parameter of water quality exceeded the limit of the legislation in the upper section of the river, while in the middle and lower sections, six and five parameters, respectively. These results show that the Sinos River presents a reduction in water quality along its extension and indicates the absence of genotoxic potential in the sampled period.

Comparative metagenome of a stream impacted by the urbanization phenomenon

Abstract Rivers and streams are important reservoirs of freshwater for human consumption. These ecosystems are threatened by increasing urbanization, because raw sewage discharged into them alters their nutrient content and may affect the composition of their microbial community. In the present study, we investigate the taxonomic and functional profile of the microbial community in an urban lotic environment. Samples of running water were collected at two points in the São Pedro stream: an upstream preserved and non-urbanized area, and a polluted urbanized area with discharged sewage. The metagenomic DNA was sequenced by pyrosequencing. Differences were observed in the community composition at the two sites. The non-urbanized area was overrepresented by genera of ubiquitous microbes that act in the maintenance of environments. In contrast, the urbanized metagenome was rich in genera pathogenic to humans. The functional profile indicated that the microbes act on the metabolism of methane, nitrogen and sulfur, especially in the urbanized area. It was also found that virulence/defense (antibiotic resistance and metal resistance) and stress response-related genes were disseminated in the urbanized environment. The structure of the microbial community was altered by uncontrolled anthropic interference, highlighting the selective pressure imposed by high loads of urban sewage discharged into freshwater environments.

Protozoa ciliates community structure in urban streams and their environmental use as indicators / Estrutura da comunidade de protozoários ciliados em córregos urbanos e sua utilização como indicadores

Abstract The objective of this work was to investigate the patterns of diversity and abundance of ciliates protozoa community in three tropical urban streams and verify which species can be considered bioindicators of water quality. In each stream, three regions (headwater, middle and mouth) were sampled in two years, in the rainy and dry seasons. The abiotic factors (dissolved oxygen, pH, temperature, turbidity, width, depth, flow and conductivity) and biotic factors (total coliforms, bacterioplankton, chlorophyll and flagellates) were analyzed using appropriate methods and ciliates were identified and counted through specialized literature. We identified 84 species distributed into 24 orders. Peniculida was the most representative order followed by Prorodontida. The RDA scores showed a spatial difference between streams. On the other hand, regarding the temporal variation, there was no separation of the sampled data. The Indval analysis showed ten species indicators, such as Coleps hirtus, Euplotes moebiusi and Tintinidium pusillum. The Cluster analysis per stream versus period showed a greater distinction of the streams in the dry season. On the other hand, a low similarity was observed between sections within the same stream. In summary, the results indicated that the ciliates can be used as important tools as bioindicators in lotic environments affected by different degrees of pollution, due to the fact that they have short life cycle, allowing the detection of impacts on a small time scale, as well as by having direct answers to environmental changes and high sensitivity to contaminants.

Resumo O objetivo deste trabalho foi investigar os padrões de diversidade e abundância da comunidade de protozoários ciliados em três córregos urbanos tropicais, bem como verificar quais espécies podem ser consideradas bioindicadoras da qualidade da água. Em cada córrego foram amostradas três regiões (cabeceira, intermediária e foz) em dois anos, nas estações chuvosa e seca. Os fatores abióticos (oxigênio dissolvido, pH, temperatura, turbidez, largura, profundidade, fluxo e condutividade) e fatores bióticos (coliformes totais, bacterioplâncton, clorofila e flagelados) foram analisados ​​por metodologia específica e os ciliados foram contados e identificados por meio de literatura especializada. Foram identificadas 84 espécies distribuídas em 24 ordens. Peniculida foi a ordem mais representativa, seguida por Prorodontida. Os escores da RDA mostraram uma diferença espacial entre os córregos. Por outro lado, em relação à variação temporal, não houve separação entre os dados amostrados. A análise Indval mostrou dez espécies indicadoras, como Coleps hirtus, Euplotes moebiusi e Tintinidium pusillum. A análise de Cluster realizada por riacho versus período evidenciou uma maior distinção dos córregos no período seco. Por outro lado, foi observada uma baixa similaridade entre os locais dentro do mesmo córrego. Em síntese, os resultados indicaram que os ciliados podem ser utilizados como ferramentas importantes como bioindicadores em ambientes lóticos afetados por diferentes graus de poluição, devido ao fato de apresentarem um curto ciclo de vida, permitindo a detecção de impactos em uma pequena escala de tempo, bem como por apresentarem respostas diretas às mudanças ambientais e alta sensibilidade a contaminantes.

Plesiomonas shigelloides exports a lethal cytotoxic-enterotoxin (LCE) by membrane vesicles

ABSTRACT Plesiomonas shigelloides isolated from water in Brazil was previously described as a hemorrhagic heat-labile cytotoxic-enterotoxin producer. We purified this toxin from culture supernatants using ion metallic affinity chromatography (IMAC) followed by molecular exclusion chromatography. The pure toxin presented molecular mass of 50 kDa and isoelectric point (pI) around 6.9 by 2D electrophoresis. When injected intravenously, the purified cytotoxic-enterotoxin induced also severe spasms followed by sudden death of mice. Hence, we entitled it as lethal cytotoxic-enterotoxin (LCE). The presence of membrane vesicles (MVs) on cell surfaces of P. shigelloides was observed by scan electron microscopy (SEM). From these MVs the LCE toxin was extracted and confirmed by biological and serological assays. These data suggest that P. shigelloides also exports this cytotoxic-enterotoxin by membrane vesicles, a different mechanism of delivering extra cellular virulence factors, so far not described in this bacterium.

Occurrence and toxicity of Microcystis aeruginosa (Cyanobacteria) in the Paraná River, downstream of the Yacyretá dam (Argentina) / Presencia y toxicidad de Microcystis aeruginosa (Cianobacteria) en el río Paraná, aguas abajo de la represa Yacyretá (Argentina)

AbstractCyanobacteria constitute the main toxin producers in inland water ecosystems and have extensive global distribution. The presence of hepatotoxins in aquatic environments is hazardous to human and animal health; even though the presence and identification of hepatotoxic microcystins in rivers and reservoirs of the world have been confirmed by several studies in the last few years. Herein, we studied the abundance and toxicity of Microcystis aeruginosa in the Argentine section of the Paraná River at the beginning of the Middle Paraná (Corrientes Hydrometer), near Corrientes city (27º28´ S - 58º51´ W) and approximately 220 km downstream of the Yacyretá dam (High Paraná). The Paraná River basin, with a drainage area of 3.1 x 106 km2 and 3 965 km in length, is the second largest catchment of South America, after that of the Amazon. The Paraná River is the main source of drinking water supply for the Northeastern Argentine region. Phytoplankton samples were collected and environmental variables were measured in a monthly basis (exceptionally fortnightly), from March 2004 to June 2008. Fifty-eight samples were analyzed for phytoplankton density and biomass. Five samples were used for toxicity testing; the latter were obtained during the cyanobacteria blooms from 2005 to 2008. Phytoplankton counts were performed with an inverted microscope, and biomass was expressed as biovolume. Bioassays with mice and high-performance liquid chromatography (HPLC) analysis were performed to evaluate the presence of cyanotoxins. Phytoplankton mainly consisted of Cryptophyta, Chlorophyta and Bacillariophyta. Microcystis aeruginosa was identified during the warmer months each year (November to March). Density varied between 189 and 25 027 cells/mL (1-10 colonies/mL) and biomass from 0.34 to 44 mm3/L. Taking into account the number of cells, the highest abundance occurred in April 2004 (25 027 cells/mL), coinciding with the largest biovolume (44 mm3/L). All mice subjected to intraperitoneal injections with samples obtained during bloom episodes showed positive results for the presence of hepatotoxins. Three microcystins variants: LR, RR and [D-Leu1] Mcyst-LR were detected by analysis with semi-preparative high-performance liquid chromatography with diode array detector system (HPLC-PDA). This constitutes the first report of microcystins recorded during M. aeruginosa blooms in the Argentine stretch of the Paraná River at the beginning of the Middle Paraná (Corrientes Hydrometer), approximately 220 km downstream of the Yacyretá dam (High Paraná).

ResumenLas Cyanobacterias constituyen el principal productor de toxinas en ecosistemas acuáticos y tienen una amplia distribución mundial. La presencia e identificación de microcistinas hepatotóxicas en ríos y embalses de todo el mundo fue confirmada por diferentes estudios durante los últimos años. La presencia de hepatotoxinas en cuerpos de agua son riesgosas para la salud humana y animal. Se estudió la abundancia y toxicidad de Microcystis aeruginosa (Kütz.) Kütz. en el río Paraná (Argentina), cerca de la ciudad de Corrientes (27°28' S - 58°51' W), aproximadamente a 220 km aguas abajo de la represa Yacyretá. La cuenca del río Paraná, con un área de drenaje de 3.1 x 106 km2 y 3 965 km de longitud, es la segunda mayor cuenca de Sudamérica, después del Amazonas. El río Paraná es la principal fuente de abastecimiento de agua potable para el Nordeste de la República Argentina. Los muestreos se realizaron mensualmente (excepcionalmente fueron quincenales) con medición de variables ambientales, entre Marzo 2004 y Junio 2008. Se tomaron un total de 58 muestras para analizar la densidad y biomasa del fitoplancton; mientras que cinco muestras fueron utilizadas en ensayos de toxicidad, estas últimas fueron obtenidas durante floraciones de cianobacterias entre 2005 y 2008. Los recuentos de fitoplancton fueron realizados con un microscopio invertido y la biomasa fue expresada como biovolumen. Para determinar la presencia de cianotoxinas se utilizaron bioensayos con ratones y análisis con Cromatografia líquida de alta resolución (HPLC). El fitoplancton estuvo representado principalmente por Cryptophyta, Chlorophyta y Bacillariophyta. Cyanobacteria fue dominante durante los meses cálidos de cada año (Noviembre a Marzo), con alta densidad de Microcystis aeruginosa. La densidad de M. aeruginosa varió entre 189 y 25 027 cells/mL (1-10 colonies/mL) y la biomasa entre 0.34 y 44 mm3/L. Teniendo en cuenta el número de células, la mayor abundancia ocurrió en abril 2004 (25 027 cells/ mL), coincidiendo con el gran biovolumen (44 mm3/L). Todos los ratones inyectados intraperitonealmente presentaron síntomas correspondientes a hepatotoxicidad. Tres variantes de microcystinas: LR, RR y [D-Leu1] Mcyst-LR, fueron detectadas por análisis de cromatografía líquida de alta resolución con detector de diodos (HPLC-PDA). Este es el primer trabajo de microcistinas registradas durante las floraciones de M. aeruginosa en el tramo argentino del río Paraná en los inicios del Paraná Medio (Hidrómetro Corrientes), aproximadamente a 220 km aguas abajo de la represa de Yacyretá (Alto Paraná).

Monitoring of metals, organic compounds and coliforms in water catchment points from the Sinos River basin / Monitoramento de metais, compostos orgânicos e coliforms em pontos de captação de água da bacia hidrográfica do Rio dos Sinos

Unplanned use and occupation of the land without respecting its capacity of assimilation and environmental purification leads to the degradation of the environment and of water used for human consumption. Agricultural areas, industrial plants and urban centres developed without planning and the control of effluent discharges are the main causes of water pollution in river basins that receive all the liquid effluents produced in those places. Over the last decades, environmental management has become part of governmental agendas in search of solutions for the preservation of water quality and the restoration of already degraded resources. This study evaluated the conditions of the main watercourse of the Sinos River basin by monitoring the main physical, chemical and microbiological parameters described in the CONAMA Resolution no. 357/2005.The set of parameters evaluated at five catchment points of water human consumption revealed a river that has different characteristics in each reach, as the upper reach was class 1, whereas the middle and lower reaches of the basin were class 4. Monitoring pointed to households as the main sources of pollutants in those reaches, although metals used in the industrial production of the region were found in the samples analyzed.

O uso e ocupação do solo não planejados, sem respeitar sua capacidade de assimilação e de purificação ambiental, levam à degradação do ambiente e da água utilizada para consumo humano. Áreas agrícolas, plantas industriais e centros urbanos desenvolvidos sem planejamento e controle de descargas de efluentes são as principais causas de poluição da água em bacias hidrográficas que recebem todos os efluentes líquidos produzidos naqueles lugares. Nas últimas décadas, gestão ambiental tornou-se parte das agendas governamentais em busca de soluções para a preservação da qualidade da água e a restauração de recursos já degradadas. Este estudo avaliou as condições do curso de água principal da bacia do Rio dos Sinos, monitorando o principal físico, químico e microbiológicos parâmetros descritos na Resolução CONAMA n.º 357/2005. O conjunto de parâmetros avaliados em cinco pontos de captação de água para consumo humano revelou um rio que tem características diferentes em cada alcance, como o superior chegar era classe 1, Considerando que o atinge médio e inferior da bacia foram classe 4. monitoramento apontado famílias como as principais fontes de poluentes naqueles atinge, apesar de metais utilizados na produção industrial da região foram encontrados nas amostras analisadas.

The Effect of Ionizing Radiation on Multi-drug Resistant Pseudomonas aeruginosa Isolated from Aquatic Environments in Egypt.

Aims: This study was conducted to determine the effect of different doses of gamma radiation on Multi-drug resistant Pseudomonas aeruginosa isolated from River Nile at Rosetta branch and associated drains in Egypt. Place and Duration of Study: The study was started with samples collection in August 2010 through April 2011 in the Microbiology Dep., Central Laboratory for Environmental Quality Monitoring (CLEQM), National Water Research Center (NWRC), Cairo, Egypt and the National Centre for Radiation Research and Technology (NCRRT), Egyptian Atomic Energy Authority (EAEA), Cairo, Egypt. Methodology: Water samples were processed using membrane filtration, 144 strains of P. aeruginosa were isolated and identified and their antibiotic susceptibility was determined against 20 different antibiotics using agar disc diffusion method. Irradiation of bacterial isolates was processed using gamma irradiation unit of cobalt (Co60) and the D10-value was calculated from the survival curve. Results: Isolates were categorized as multi-drug resistant Pseudomonas aeruginosa (MDRPA). 125 (86.8%) were found to be extensively drug resistant (XDR) and 19 (13.2 %) were characterized as possible pan drug resistant (PDR). The highest resistance (100%) was mostly directed to amoxycillin/clavulanic acid, ampicillin, carbenicillin, methicillin, cephalothin, kanamycin, vancomycin, tetracycline, erythromycin, clindamycin, trimethoprim/sulfamethoxazole, nitrofurantoin and chloramphenicol. More than 75% of isolates were sensitive to norfloxacin (82.6%), piperacillin (81.2%), amikacin (79.2%) and tobramycin (77.8%). 63.2%, 26.4% and 14.6% of isolates were sensitive to ofloxacin, cefotaxime and ceftriaxone, respectively. The viable counts of MDRPA decreased with increasing radiation doses of gamma rays up to the lethal dose (3 kGy). The counts of 0.5, 1.0, 1.5, 2.0, 2.5 kGy irradiated samples were respectively 7.8, 6.5, 4.7, 2.3& 1 log10 and the D10-value calculated from the survival curve was 0.27 kGy. Conclusion: Contaminated fresh water may act as reservoirs for antibiotic resistant pathogens. Regular monitoring of Multi-drug resistant pathogens in aquatic environments should be adopted constantly. Gamma radiation demonstrates a potential value for wastewater treatment and pollution control.

Comunidade fitoplanctônica no monitoramento de rios do estado de São Paulo / Phytoplankton community in monitoring rivers of São Paulo state

Objetivo. São poucos os estudos desenvolvidos sobre o fitoplâncton que habita os rios. No estado de São Paulo muitas das captações de água para o abastecimento público se localizam em rios de planície ou rios de baixo gradiente, e apresentam uma grande quantidade de barragens com reservatórios construídos no seu curso, além da presença de lagoas marginais e de pequenos represamentos para a captação de água, contribuindo para o desenvolvimento dos organismos planctônicos. Assim frente à necessidade de conhecer melhor a estrutura e a dinâmica de algas e cianobactérias planctônicas em rios visando aprimorar sua utilização como ferramenta de monitoramentos, o objetivo principal deste projeto foi o de reavaliar e discutir melhorias na aplicação do índice da comunidade fitoplânctonica (ICF), desenvolvido e utilizado pela CETESB, nestes ambientes. Metodologia. Foram avaliados oito locais de coleta, em rios junto a captações de água para o abastecimento público de seis importantes municípios do estado, durante o ano de 2009. As amostragens foram bimestrais e foram levantados os aspectos abióticos (precipitação, vazão, temperatura, pH, oxigênio dissolvido, turbidez, condutividade, carga orgânica e nutrientes) e biológicos (Coliformes termotolerantes, Clorofila a e a Comunidade Fitoplanctônica). Foram testadas e propostas alterações no ICF para aplicação em rios e aplicados outros índices de qualidade das águas utilizados pela CETESB como IET, IVA, ICF para comparação. Resultados. A comunidade fitoplânctônica nos rios estudados apresentou uma elevada riqueza de espécies, porém com uma alta proporção de espécies raras. A densidade dos organismos foi considerada baixa para a maioria dos rios. As maiores densidades estiveram mais relacionadas com o aumento da carga orgânica e as formas de nitrogênio, com aumento significativo no período de menor precipitação. O grupo Baccilariophyta, principalmente a classe Coscinodiscophyceae estiveram associados aos rios com melhor qualidade da água, enquanto a classe Chlorophyceae da ordem Chlorococcales foi correlacionada com o aumento da carga orgânica. A classe Cyanobacteria predominou nos rios com os maiores valores de fósforo e coliformes termotolerantes. A classe Cryptophyceae teve seu aumento relativo relacionado principalmente ao período mais seco com o decréscimo de outros grupos.

Cyanobacteria abundance and its relationship to water quality in the Mid-Cross River floodplain, Nigeria

The physicochemical variables and cyanobacteria of Mid-Cross River, Nigeria, were studied in six stations between March 2005 and August 2006 to determine the relationship between water quality and cyanobacteria abundance. Canonical Correspondence Analysis (CCA) showed that biological oxygen demand (BOD), dissolved oxygen, pH, water velocity, width and depth were important environmental factors that influenced cyanobacteria abundance. Trace metals, phosphate and nitrate increased significantly from values of previous studies indicating increased eutrophication of the river but were weakly correlated with cyanobacteria abundance and could be scarcely regarded as regulating factors. A higher cyanobacteria abundance was recorded during the wet season in most of the sampled stations. The dominant cyanobacteria included Microcystis aeruginosa, Aphanizomenon flos-aquae, Oscillatoria limnetica and Anabaena spiroides. The toxins produced by these species could degrade water quality. The factors favouring cyanobacteria abundance were identified as increased pH, width and depth. Increase in cyanobacteria abundance was associated with reduction in dissolved oxygen and increase in BOD values. Rev. Biol. Trop. 57 (1-2): 33-43. Epub 2009 June 30.

Las variables físico-químicas y la abundancia de cianobacterias del río nigeriano Mid-Cross fueron estudiadas en seis estaciones entre marzo del 2005 y agosto del 2006. El Análisis de Correspondencia Canónica (CCA) demostró que la demanda biológica de oxígeno (DBO), oxígeno disuelto, pH, velocidad de agua, anchura y profundidad son factores ambientales importantes que influyen en la abundancia de cianobacterias. Los mayores valores de trazas de metales, fosfatos y nitratos, en comparación con estudios previos, indican mayor eutrofización, pero tienen poca correlación con la abundancia de las cianobacterias. La mayor abundancia de cianobacterias se registró durante el periodo más húmedo de la estación seca en la mayoría de las estaciones de muestreo. Las cianobacterias dominantes incluyen Microcystis aeruginosa, Aphanizomenon flos-Aquae, Oscillatoria limnetica y Anabaena spiroide. Las toxinas producidas por estas especies podrían degradar la calidad del agua. Los factores que favorecen la abundancia de cianobacterias fueron identificados como el aumento de pH, ancho y profundidad. El aumento de la abundancia se asoció con una reducción de oxígeno disuelto y un aumento de los valores de DBO.

Effects of bacterial infestation caused by human wastes on the skin structures of Mugil platanus Günther, 1880 (Mugilidae) / Efeitos da infestação de bactérias provocada por resíduos humanos na pele de Mugil platanus Günther, 1880 (Mugilidae)

Manifestation of infectious pathologies in fishes usually increases in environments where organic wastes are disposed. Specimens of Mugil platanus Günther, 1880 and water samples collected at three points of the Tramandaí river were analyzed during a one year period. The macroscopic observation revealed ulcerations in the caudal peduncle area covered with a mass of amorphous and whitened tissues. Histopathologic analysis showed the presence of negative gram bacteria, probably responsible for alterations of the normal structure of the epidermic tissues. Non-parametric statistical analysis for ammonia concentration showed a significant variation among the three collected spots as well as in the multiple comparison between two spots. In this study, we describe cutaneous lesions observed in Mugil platanus specimens and tested their correlation with environmental ammonia concentration.

A manifestação de patologias infecciosas em populações de peixes costuma ser incrementada em ambientes onde são lançados despejos orgânicos. Exemplares da espécie Mugil platanus Günther, 1880 coletados em três pontos do rio Tramandaí foram utilizados para análise das alterações, e amostras de água foram tomadas durante o período de um ano para verificação da presença de bactérias. A observação macroscópica revelou ulcerações na região do pedúnculo caudal revestidas por uma massa de tecido amorfo e esbranquiçado. A análise histopatológica mostrou a presença de bactérias gram negativas, que provavelmente foram as responsáveis pela alteração da estrutura normal dos tecidos de revestimento do corpo. O resultado da análise estatística não paramétrica para a concentração de amônia mostrou variação significativa entre os três pontos amostrados bem como nas comparações múltiplas entre os pontos dois a dois. Este trabalho descreve as lesões cutâneas observadas em Mugil platanus e testa sua correlação com a concentração de amônia.

Pseudomonas aeruginosa: disseminação de resistência antimicrobiana em efluente hospitalar e água superficial / Pseudomonas aeruginosa: spread of antimicrobial resistance in hospital effluent and surface water

O objetivo deste estudo foi comparar amostras de efluente do Hospital São Vicente de Paulo com amostras de água do Rio Passo Fundo, quanto ao perfil de susceptibilidade de isolados de Pseudomonas aeruginosa, para inferir sobre a presença de isolados de origem hospitalar em amostras de água superficial. A significância estatística entre os perfis de susceptibilidade das amostras foi testada por análise de variância e a comparação das amostras foi feita por contrastes de interesse. Foram identificados 198 isolados de Pseudomonas aeruginosa a partir das amostras analisadas. O fenótipo de multirresistência não foi observado nas amostras do Rio Passo Fundo, embora alguns isolados resistentes a carbapenêmicos tenham sido identificados, indicando a presença de contaminação com bactérias provenientes de um ambiente sob forte pressão seletiva. Diferenças significativas entre as amostras de água e efluente hospitalar foram observadas a partir da análise de variância por contrastes de interesse.

The aim of this study was to compare sewage samples from Hospital São Vicente de Paulo with water samples from the Passo Fundo river, with regard to the susceptibility profile of Pseudomonas aeruginosa isolates, in order to make inferences about the presence of strains of hospital origin in surface water samples. The statistical significance between the susceptibility profiles of the samples was tested using analysis of variance, and the samples were compared by means of contrasts of interest. One hundred and ninety-eight isolates of Pseudomonas aeruginosa were recovered from the samples analyzed. No phenotype for multiresistance was found in the samples from the Passo Fundo river, although some carbapenem-resistant isolates were identified, thereby indicating the presence of contamination with bacteria derived from an environment under strong selection pressure. Significant differences between the water and hospital effluent samples were observed, based on the analysis of variance by means of contrasts of interest.

Detection of viable and viable nonculturable Vibrio cholerae O1 through cultures and immunofluorescence in the Tucumán rivers, Argentina

Vibrio cholerae has been sporadically isolated from rivers in Tucumán, Argentina, since the outbreak in 1991. The aim of this study was to determine the environmental reservoir of the bacterium in these rivers, assessing the presence of Vibrio cholerae non-O1 and O1 (the latter both in its viable culturable and non culturable state) and its relationship to environmental physicochemical variables. 18 water samplings were collected in the Salí River (in Canal Norte and Banda) and the Lules River between 2003 and 2005. Physical-chemical measurements (pH, water temperature, electrical conductivity and dissolved oxygen) were examined. Vibrio cholerae was investigated with conventional culture methods and with Direct Immunofluorescence (DFA-VNC) in order to detect viable non culturable organisms. All isolated microorganisms corresponded to Vibrio cholerae non-O1 and non-O139 (Lules 26 percent, Canal Norte 33 percent and Banda 41 percent). The majority was found during spring and summer and correlated with temperature and pH. Non culturable Vibrio cholerae O1 was detected year round in 38 of the 54 water samples analyzed. Application of the Pearson correlation coefficient revealed that there was no relationship between positive immunofluorescence results and environmental physicochemical parameters. Genes coding for somatic antigen O1 were confirmed in all DFA-VNC-positive samples, whereas the virulence-associated ctxA and tcpA genes were confirmed in 24 samples.

Vibrio cholerae tem sido isolado esporadicamente nos rios da Província de Tucumán, Argentina, desde outubro de 1991. O objetivo deste estudo foi localizar os reservatórios nestes rios, identificar a presença de Vibrio cholerae O1 (em estado cultivável e não cultivável) e relacionar a presença desta bactéria com as variações físico-químicos da água. Foram coletadas dezoito amostras de água do rio Salí (nas localidades de Canal Norte e Banda) e do rio Lules, entre 2003 e 2005. Estas foram submetidas a análises físico-químicos como determinação de pH, temperatura, condutibilidade elétrica e oxigênio dissolvido. A presença de Vibrio cholerae foi verificada por métodos de cultivo convencional e por imunofluorescência direta (DFA-VNC). Todos os microrganismos isolados foram não O1 e não O139 (Lules 26 por cento, Canal Norte 33 por cento e Banda 41 por cento). A maioria foi encontrada na primavera e verão, indicando uma relação com a temperatura e pH. Das 54 amostras analisadas por DFA-VNC, 38 Vibrio cholerae não cultivável, foram detectadas em todas as épocas do ano. As amostras positivas foram confirmadas por PCR para o antígeno somático O1 e para os genes de virulência ctxA e tcpA. Coeficiente de correlação de Pearson revelou que não há relação entre os resultados obtidos por imunofluorescência e a variação dos parâmetros físico-químicos.

Aspectos limnológico-sanitários das águas do rio Caulim na Região Metropolitana de São Paulo / Sanitary and limnologic aspects of the water from the Caulim river, in the metropolitan region of São Paulo

Introdução: A água é um recurso essencial para a vida dos seres vivos e tem sofrido grande deterioração devido principalmente, à ação antrópica. O Rio Caulim está situado no sub-distrito de Parelheiros, onde a ocupação se deu de forma descontrolada e inadequada e vem recebendo o esgoto doméstico dos loteamentos clandestinos da região. Como deságua na Represa do Guarapiranga, importante manancial de abastecimento da Região Metropolitana de São Paulo, seu estudo torna-se importante. Objetivo: Analisar as condições ecológico-sanitárias do Rio Caulim; avaliar parâmetros físicos, químicos e microbiológicos do Rio Caulim; verificar a variação temporal e espacial desses parâmetros; conhecer as possíveis relações da qualidade da água com a ocupação do solo; e fornecer subsídios para as autoridades tomarem medidas de recuperação e preservação da qualidade das águas do Rio Caulim. Métodos: Em seis pontos ao longo do rio foram analisadas variáveis abióticas (temperatura do ar e da água, profundidade, transparência, oxigênio dissolvido, pH, condutividade, cor, turbidez, sólidos totais e suspensos, demanda bioquímica de oxigênio, nitrogênio total e amoniacal, fósforo total e ortofosfato total) e variáveis bióticas (coliformes totais e termotolerantes). Essas foram analisadas quanto à variação espaço/temporal e realizou-se o teste de correlação de Spearmam para verificar a relação existente entre as variáveis. Resultados: Os valores encontrados para condutividade (182 a 783 µS), turbidez (11 a 160 UNT), demanda bioquímica de oxigênio (5 a 202 mg/L O2), nitrogênio total (7 a 93 mg/L N), nitrogênio amoniacal (2 a 36 mg/L N), fósforo total (0,1 a 9 mg/L P), orto-fosfato (0,1 a 5 mg/L P), coliformes totais (10 elevado a quinta potencia a 10 elevado a sétima potencia NMP) e coliformes termotolerantes ( 10 elevado à quarta potencia a 10 elevado a sétima potencia NMP), foram altos valores encontrados para oxigênio dissolvido (0,4 a 5,6 mg/L O2) foram baixos.Os pontos apresentaram...

Development of water quality test kit based on substrate utilization and toxicity resistance in river microbial communities

Methods for measuring toxicity or respiratory activity of microbial cultures can be used as tools for assessing the presence of chemicals and their impact on the streams. The proposed toxicity test is based on the respirometric characteristics of the bacteria according to the principals of Biolog's microplate system. As the microorganism are utilizing the carbon source [peptone], the reduction of the tetrazolium dye as the redox indicator is taking place, leading to a developing change in the well's color The tests provided us with information in regard to the toxicity range of the chemicals with activated sludge and Alcaligenes