Maria do Carmo Bittencourt-Oliveira

Detection of potential microcystin-producing cyanobacteria in Brazilian reservoirs with a mcyB molecular marker

One of the most serious problems related to water eutrophication is the occurrence of increasingly frequent blooms of toxic cyanobacteria in freshwater ecosystems. Microcystin (MCYST) molecular markers may be used for the detection of toxic cyanobacteria, both cultivated strains and environmental samples, independently of their taxonomic category and production of the toxin at the moment of analysis. Sixty Microcystis spp. strains from 15 water reservoirs of south, southeastern and northeastern Brazil were analyzed by polymerase chain reaction (PCR) with oligonucleotide primers for mcyB gene of the operon that encodes a microcystin synthetase. It was found out that the presence of a unique amplified product of approximately 780 bp in 18 strains, indicated the presence of the microcystin-producing genotype. There was correspondence between the presence of the mcyB gene and microcystin determined by ELISA. Eight reservoirs contained toxic strains, two of these reservoirs being used mainly for public water supply. The coexistence of a mixture of toxic and non-toxic genotypes in populations of several reservoirs was found. Thus, it is evident that Microcystis populations present in blooms compose a mosaic, with genetically different individuals within the same population, each one, possibly, with its own tolerance to environmental factors and with distinct toxicity potential.

Temporal variation of the phytoplankton community at short sampling intervals in the Mundaú reservoir, Northeastern Brazil

4µg.L -1 , sendo que as cianobacterias apresentaram valores de biomassa entre 1,07×10 4 e 8,21×10 4 µg.L -1 . A mais alta disponibilidade de fosforo, limitacao de nitrogenio, pH alcalino e estabilidade termica favoreceram floracoes de cianobacterias, especialmente durante o periodo seco. As diferencas entre os horarios e as estacoes amostrais na distribuicao da comunidade fitoplanctonica foram determinadas pelos valores de temperatura, pH, fosforo total e turbidez. As associacoes fitoplanctonicas que mais se destacaram foram do grupo funcional Sn, formado por Cylindrospermopsis raciborskii, que predominou no sistema em ambos periodos sazonais. Concluiu-se que o fitoplâncton apresentou uma forte influencia das variaveis abioticas especialmente no estabelecimento de padroes sazonais de distribuicao. Palavras-chave: Cylindrospermopsis raciborskii, eutrofizacao, nutrientes, reservatorio de abastecimento

Bioaccumulation of Microcystins in Lettuce

The contamination of lettuce (Lactuca sativa L.) by water‐borne crude extracts of the cyanobacterium microcystin‐producing Microcystis aeruginosa (Kützing) Kützing was investigated. The aim of the study was to determine whether bioaccumulation of microcystins occurs in lettuce foliar tissue when sprayed with solutions containing microcystins at concentrations observed in aquatic systems (0.62 to 12.5 μg · L−1). Microcystins were found in lettuce foliar tissues (8.31 to 177.8 μg per Kg of fresh weight) at all concentrations of crude extracts. Spraying with water containing microcystins and cyanobacteria may contaminate lettuce at levels higher than the daily intake of microcystins recommended by the World Health Organization (WHO), underscoring the need to monitor such food exposure pathways by public authorities.

Phytoplanktonic associations: a tool to understanding dominance events in a tropical Brazilian reservoir

ABSTRACT – (Phytoplanktonic associations: a tool to understanding dominance events in a tropical Brazilian reservoir). The aim ofthis study was to characterize phytoplankton associations, as well as discuss controlling factors determining algal dominance in aeutrophic tropical reservoir, Mundau, Pernambuco, Brazil. Water samples were collected during the dry period (January/2005) and therainy period (June/2005). The samples were collected from both limnetic and littoral regions, and the phytoplankton assemblagesidentified from current literature after preservation in formaldehyde 4%. At the same time as sampling was done, in situ measurementsof water temperature, transparency, dissolved oxygen, and pH were also taken. Total phosphorus, total nitrogen concentration an d theTrophic State Index were subsequently determined in the laboratory. Phytoplankton density (ind. L -1 ) was estimated using an invertedZeiss microscope. Grouping of the phytoplankton associations was carried out using the Reynolds phytosociological classification.During the dry period, reservoir water showed low dissolved oxygen concentrations, alkaline pH and was relatively turbid compar ed tothe situation during the rainy season. Reservoir water is limited by nitrogen during both seasonal periods. The Trophic State Index isclassified as determining eutrophic conditions. Phytoplankton was represented by 70 infrageneric taxa grouped in 16 functional associations,with the majority typical of eutrophic systems. This fact is supported by quantitative analysis, which shows the dominance of

Cyanobacterial blooms in stratified and destratified eutrophic reservoirs in semi-arid region of Brazil

This study investigated the dynamics of cyanobacteria in two deep, eutrophic reservoirs in a semi-arid region of Brazil during periods of stratification and destratification. Four collections were carried out at each reservoir at two depths at three-month intervals. The following abiotic variables were analyzed: water temperature, dissolved oxygen, pH, turbidity, water transparency, total phosphorus, total dissolved phosphorus, orthophosphate and total nitrogen. Phytoplankton density was quantified for the determination of the biomass of cyanobacteria. The data were analyzed using CCA. Higher mean phytoplankton biomass values (29.8 mm(3).L(-1)) occurred in the period of thermal stratification. A greater similarity in the phytoplankton communities also occurred in this period and was related to the development of cyanobacteria, mainly Cylindrospermopsis raciborskii (>3.9 mm(3).L(-1)). During the period of thermal destratification, this species co-dominated the environment with Planktothrix agardhii, Geitlerinema amphibium, Microcystis aeruginosa and Merismopedia tenuissima, as well as with diatoms and phytoflagellates. Environmental instability and competition among algae hindered the establishment of blooms more during the mixture period than during the stratification period. Thermal changes in the water column caused by climatologic events altered other physiochemical conditions of the water, leading to changes in the composition and biomass of the cyanobacterial community in tropical reservoirs.

Cyanobacteria, microcystins and cylindrospermopsin in public drinking supply reservoirs of Brazil

Brazil has a history of blooms and contamination of freshwater systems by cyanobacterial toxins. The monitoring relevance of toxins from cyanobacteria in reservoirs for public supply is notorious given its high toxicity to mammals, included humans beings. The most recurrent toxins in Brazilian water bodies are microcystins (MC). However, the recent record of cylindrospermopsin (CYN) in northeastern Brazil, Pernambuco state, alerts us to the possibility that this could be escalating. This study reports occurrence of MC and CYN, quantified with ELISA, in 10 reservoirs, devoted to public drinking supply in northeastern Brazil. The composition and quantification of the cyanobacteria community associated with these water bodies is also presented. From 23 samples investigated for the presence of MC, and CYN, 22 and 8 out were positive, respectively. Considering the similarity of the cyanobacteria communities found in reservoirs from Pernambuco, including toxin-producing species associated to MC and CYN, we suggest that geographic spreading can be favored by these factors. These issues emphasize the need for increased monitoring of MC and CYN in drinking supply reservoirs in Brazil.

Saxitoxins accumulation by freshwater tilapia (Oreochromis niloticus) for human consumption

The accumulation of saxitoxins (STXs) in fish from freshwater aquaculture was investigated for the first time in the present study. Cyanotoxins have been monitored in liver and muscle samples of Oreochromis niloticus by chromatographic methods, both before and after the depuration process. The results show that tilapia can accumulate STXs. Our findings suggest that depuration with clean water is an alternative process to eliminate STXs from fish and, therefore, improve the safety of tilapia for consumers.

Lettuce irrigated with contaminated water: Photosynthetic effects, antioxidative response and bioaccumulation of microcystin congeners

The use of microcystins (MCs) contaminated water to irrigate crop plants represents a human health risk due to their bioaccumulation potential. In addition, MCs cause oxidative stress and negatively influence photosynthetic activities in plants. The present study was aimed at investigating the effect of MCs on photosynthetic parameters and antioxidative response of lettuce. Furthermore, the bioaccumulation factor (BAF) of total MCs, MC-LR and MC-RR in the vegetable after irrigation with contaminated water was determined. Lettuce crops were irrigated for 15 days with water containing cyanobacterial crude extracts (Microcystis aeruginosa) with MC-LR (0.0, 0.5, 2.0, 5.0 and 10.0 µg L(-1)), MC-RR (0.0, 0.15, 0.5, 1.5 and 3.0 µg L(-1)) and total MCs (0.0, 0.65, 2.5, 6.5 and 13.0 µg L(-1)). Increased net photosynthetic rate, stomatal conductance, leaf tissue transpiration and intercellular CO2 concentration were recorded in lettuce exposed to different MCs concentrations. Antioxidant response showed that glutathione S-transferase activity was down-regulated in the presence of MCs. On the other hand, superoxide dismutase, catalase and peroxidase activities were upregulated with increasing MCs concentrations. The bioaccumulation factor (BAF) of total MCs and MC-LR was highest at 6.50 and 5.00 µg L(-1), respectively, while for MC-RR, the highest BAF was recorded at 1.50 µg L(-1) concentration. The amount of total MCs, MC-LR and MC-RR bioacumulated in lettuce was highest at the highest exposure concentrations. However, at the lowest exposure concentration, there were no detectable levels of MC-LR, MC-RR and total MCs in lettuce. Thus, the bioaccumulation of MCs in lettuce varies according to the exposure concentration. In addition, the extent of physiological response of lettuce to the toxins relies on exposure concentrations.

Study of environmental burden of lead in children using teeth as bioindicator

The south region of Sao Paulo city hosts the Guarapiranga dam, responsible for water supply to 25% of the city population. Their surroundings have been subject to intense and irregular occupation by people from very low socioeconomics classes. Measurements undertaken on sediment and particulate materials in the dam revealed concentrations of lead, copper, zinc and cadmium above internationally accepted limits. Epidemiological and toxicological studies undertaken by the World Health Organization in individuals exhibiting lead concentrations in blood, near or below the maximum recommended (10 microg dl(-1)), surprisingly revealed that toxic effects are more intense in individuals belonging to low socioeconomics classes. Motivated by these facts, we aimed at the investigation of chronic incorporation of lead, as well as the use of our BIOKINETICS code, which is based on an accepted ICRP biokinetics model for lead, in order to extrapolate the results from teeth to other organs. The focus of our data taking was children from poor families, living in a small, restrict and allegedly contaminated area in São Paulo city. Thus, a total of 74 human teeth were collected. The average concentration of lead in teeth of children 5 to 10 years old was determined by means of a high-resolution inductively coupled plasma mass spectrometer (ICP-MS). For standardization of the measurements, an animal bone certified material (H-Animal Bone), from the International Atomic Energy Agency, was analyzed. The amount of lead in children living in the surroundings of the dam, was approximately 40% higher than those from the control region, and the average lead concentration was equal to 1.3 microg g(-1) approximately. Grouping the results in terms of gender, tooth type and condition, it was concluded that a carious molar of boys is a much more efficient contamination pathway for lead, resulting in concentrations 70% higher than in the control region. We also inferred the average concentrations of lead in other organs of these children, by making use of our BIOKINETIC code.

Microcystin‐producing genotypes from cyanobacteria in Brazilian reservoirs

The aim of this study was to evaluate the use of new oligonucleotide primers (mcyB‐F/R, mcyB‐F/R‐A, and mcyB‐F/R‐B) designed from Brazilian cyanobacteria for the detection of microcystin‐producing genotypes in 27 environmental samples from water reservoirs and 11 strains of Microcystis. Microcystins were found using HPLC in all 11 strains and 19 of the environmental samples. The new oligonucleotide primers amplified fragments of microcystin‐producing genes, including the eight environmental samples in which no microcystins were detected by HPLC, but which presented amplified fragments, thereby demonstrating the existence of microcystin‐producing genes. The new oligonucleotide primers exhibited better specificity when used with environmental samples and were more reliable in comparison with those described in the literature (mcyB‐FAA/RAA and mcyA‐Cd/FR), which generate false‐negative results. The better performance of these new oligonucleotide primers underline the need for designing molecular markers that are well fitted to the regional biological diversity. As this is a fast predictive technique for determining the presence or absence of microcystins, it could be used either alone or in conjunction with other techniques, such as the screening of samples to be sent for quantitative toxicological analysis using HPLC, thereby reducing monitoring cost and time.