Effect of temperature on microcystin-LR removal and lysis activity on Microcystis aeruginosa (cyanobacteria) by an indigenous bacterium belonging to the genus Achromobacter.

Microcystis is a frequent cyanobacterium bloom-forming with cosmopolitan distribution which can produce a hepatotoxin group called microcystins (MCs). These MCs are resistant to the traditional processes employed in the water treatment plants and they are often detected after conventional treatments. Because of this, the bio-removal studies have obtained a great interest in the last decades. In this work, a bacterial strain namely LG1 with the ability to remove microcystin-LR (MC-LR) under laboratory conditions was isolated from Rio de la Plata River and it was identified as Achromobacter spp. This ubiquitous bacterium was able to remove 79.5% MC-LR in 7 days with average removal time of 3.33 ± 0.08, 3.06 ± 0.05, and 2.77 ± 0.05 days at 28, 32, and 36 ± 1 °C, being higher at high temperature (36 °C) with an activation energy = 16.79 ± 1.99 kJ mol-1. LG1 grew better at higher temperature (from 28 to 36 ± 1 °C) increasing the specific growth rate (µ) and reducing 2-fold the lag phase duration (LPD) without significant differences (p > 0.05) between maximum population density (MPD). In addition, LG1 showed a lysis activity on two M. aeruginosa native strains in 7 days measured as chlorophyll a (Chl-a) concentration. The lysis activity increased around 2-fold when increasing the temperature from 28 to 36 ± 1 °C. This is the first report of an indigenous bacterium belonging to the genus Achromobacter spp. isolated from the Rio de la Plata River with the capacity to remove MC-LR and lysis activity on M. aeruginosa.

Physiological responses and toxin production of Microcystis aeruginosa in short-term exposure to solar UV radiation.

The aim of this study was to evaluate the effects of short-term (hours) exposure to solar UV radiation (UVR, 280-400 nm) on the physiology of Microcystis aeruginosa. Three solar radiation treatments were implemented: (i) PAR (PAR, 400-700 nm), (ii) TUVA (PAR + UVAR, 315-700 nm) and (iii) TUVR (PAR + UVAR + UVBR, 280-700 nm). Differential responses of antioxidant enzymes and the reactive oxygen species (ROS) production to UVR were observed. Antioxidant enzymes were more active at high UVR doses. However, different responses were observed depending on the exposure to UVAR or UVBR and the dose level. No effects were observed on the biomass, ROS production or increased activity of superoxide dismutase (SOD) and catalase (CAT) compared to the control when UVR + PAR doses were lower than 9875 kJ m-2. For intermediate doses, UVR + PAR doses between 9875 and 10 275 kJ m-2, oxidative stress increased while resistance was imparted through SOD and CAT in the cells exposed to UVAR. Despite the increased antioxidant activity, biomass decrease and photosynthesis inhibition were observed, but no effects were observed with added exposure to UVBR. At the highest doses (UVR + PAR higher than 10 275 kJ m-2), the solar UVR caused decreased photosynthesis and biomass with only activation of CAT by UVBR and SOD and CAT by UVAR. In addition, for such doses, a significant decrease of microcystins (MCs, measured as MC-LR equivalents) was observed as a consequence of UVAR. This study facilitates our understanding of the SOD and CAT protection according to UVAR and UVBR doses and cellular damage and reinforces the importance of UVR as an environmental stressor. In addition, our results support the hypothesized antioxidant function of MCs.

Hepatic and intestine alterations in mice after prolonged exposure to low oral doses of Microcystin-LR.

Oral intake of Microcystin-LR (MC-LR) is the principal route of exposure to this toxin, with prolonged exposure leading to liver damage of unspecific symptomatology. The aim of the present paper was therefore to investigate the liver and intestine damage generated by prolonged oral exposure to low MC-LR doses (50 and 100 µg MC-LR/kg body weight, administrated every 48 h during a month) in a murine model. We found alterations in TBARS, SOD activity and glutathione content in liver and intestine of mice exposed to both doses of MC-LR. Furthermore, the presence of MC-LR was detected in both organs. We also found hepatic steatosis (3.6 ± 0.6% and 15.3 ± 1.6%) and a decrease in intraepithelial lymphocytes (28.7 ± 5.0% and 44.2 ± 8.7%) in intestine of 50- and 100-µg MC-LR/kg treated animals, respectively. This result could have important implications for mucosal immunity, since intraepithelial lymphocytes are the principal effectors of this system. Our results indicate that prolonged oral exposure at 50 µg MC-LR/kg every 48 h generates significant damage not only in liver but also in intestine. This finding calls for a re-appraisal of the currently accepted NOAEL (No Observed Adverse Effect Level), 40 µg MC-LR/kg body weight, used to derive the guideline value for MC-LR in drinking water.

Neurobehavioural effects of exposure to fluoride in the earliest stages of rat development.

It is known that exposure to high concentrations of Fluoride (F) produces deleterious health effects in human population. However, in the last years it has been concluded that low concentrations of F may have adverse health effects as well. Transplacental passage of F and its incorporation into foetal tissues has been demonstrated. Therefore, the purpose of the present work was to study the effects of the exposure to low levels of F during pregnancy and lactation on the central nervous system functionality. Wistar rats were exposed to low F concentrations (5 and 10 mg/l) during pregnancy and lactation. Sensorimotor reflexes in the each pup were analysed and the postnatal day on which both eyes and auditory canals were opened was recorded. Locomotor activity and anxiety were subsequently analysed in 45- and 90-day-old offspring by an open field test and plus maze test, respectively. A significant delay in the development of eye opening was observed in all offspring whose mothers had been exposed to the two F concentrations tested. Exposure to 5 and 10 mg/l F was also found to significantly decrease locomotor activity only in 90-day-old male and female offspring. A low index of anxiety in the young females and in all adult offspring exposed to the two F concentrations tested was also detected. Taken together, findings from the present study show that exposure to low F concentrations during pregnancy and lactation produces dysfunction in the central nervous system mechanisms which regulate motor and sensitive development, locomotor activity and anxiety

Locomotor activity and sensory-motor developmental alterations in rat offspring exposed to arsenic prenatally and via lactation.

Arsenic (As) is one of the most toxic naturally occurring contaminants in the environment. The major source of human exposure to inorganic As (iAs) is through contaminated drinking water. Although both genotoxicity and carcinogenicity derived from this metalloid have been thoroughly studied, the effects of iAs on the development and function of the central nervous system (CNS) have received less attention and only a few studies have focused on neurobehavioral effects. Thus, in order to characterize developmental and behavioral alterations induced by iAs exposure, pregnant Wistar rats were exposed to 0.05 and 0.10 mg/L iAs through drinking water during gestation and lactation. Sensory-motor reflexes in each pup were analyzed and the postnatal day when righting reflex, cliff aversion and negative geotaxis were recorded. Functional Observational Battery (FOB) and locomotor activity in an open field were assessed in 90-day-old offspring. Results show that rats exposed to low iAs concentrations through drinking water during early development evidence a delay in the development of sensory-motor reflexes. Both FOB procedure and open-field tests showed a decrease in locomotor activity in adult rats. This study reveals that exposure to the above-mentioned iAs concentrations produces dysfunction in the CNS mechanisms whose role is to regulate motor and sensory development and locomotor activity.

Biomarkers of prolonged exposure to microcystin-LR in mice.

The effects of prolonged exposure to microcystins (MCs) on health are not yet sufficiently understood and this type of poisoning is often undiagnosed. Even though chronic exposure has been linked with liver cancer and alterations have been described in liver damage marker enzymes in exposed populations, there are not profile parameters that indicate prolonged exposure to microcystins. The aim of this work is to determine, based on an animal model of prolonged exposure to successive i.p. doses of 25 µg MC-LR/kg body weight, several plasma parameters which could be useful as exposure biomarkers. Hemoglobin (Hb) and methemoglobin (MetHb) levels were determined on blood samples. We also studied plasma levels of hydroperoxides (ROOHs), α-tocopherol, glutathione and lipid profile as well as superoxide dismutase (SOD) and catalase (CAT) erythrocyte activities. In addition, the determination of MC-LR levels in liver, kidney, plasma, urine and feces of treated mice was carried out. We found that alteration in MetHb, ROOHs, glutathione, α-tocopherol levels, SOD activity and plasma lipid profile, correlates with those expected if the alteration derived from hepatic damage. The alterated plasma paramenters together with MC-LR determination could be used as biomarkers, helpful tools in screening and epidemiological studies.

Occurrence of Microcystis aeruginosa and microcystins in Rio de la Plata river (Argentina)

This paper is the first report on microcystins producer blooms of Microcystis aeruginosa in the Argentinean coast of the R¨ªo de la Plata river, the most important drinkingwater supply of Argentina. The distribution of toxic cyanobacterium Microcystis cf. aeruginosa blooms in the Argentinean coast of the Rio de la Plata river was studied from December 2003 and January 2006. Microcystis aeruginosa persisted in the river with values ranged between 0 -7.8 104 cells ml-1. Samples of two Microcystis aeruginosa water blooms were collected at La Plata river and were analyzed by the mouse bioassay and by high-performance liquid chromatography with Diode-array and MS detector. The samples showed high hepatotoxicityin mouse bioassay and, in accordance, important amount of microcystins. The bloom samples contained microcystins LR and a variant of microcystin with a molecular ion [M+H]+= 1037.8 m/z as major components. The total toxin content found in these samples was 0.94¦Ìg/mg and 0.69¦Ìg/mg of lyophilised cells. We conclude that the presence of toxic clones of Microcystisaeruginosa in the Argentinean coast of the Rio de la Plata is an actual sanitary and environmental problem and that further studies are necessary to make the risk assessment.

Drinking water: problems related to water supply in Bahía Blanca, Argentina / Agua potable: problemas en el abastecimiento de la ciudad de Bahía Blanca, Argentina

In April 2000, alterations in drinking water quality, such as turbidity and odors similar to those of organochloride pesticides, were detected at Bahía Blanca (Argentina). This fact was associated to reports of dermic reactions and respiratory problems in the population. Samples drawn from Paso de las Piedras reservoir, two water treatment plants of at inlet and outlet points and at several particular houses were analyzed. Total phytoplankton in the reservoir varied between 49440 and 84832 cells.ml-1, while dominant species was, Anabaena circinalis. Efficiency to remove microorganisms (ER) by the treatment plants considering cell number of total phytoplankton was analyzed. Bacteriological analysis, qualitative and quantitative studies of phytoplankton, pesticides, THM (trihalomethanes) and BTEX (benzene, toluene, ethylbenzene and xylene) analysis were carried out in samples of domestic supply and in the treatment plants. Even though samples were bacteriological potable, Anabaena circinalis and Microcystis aeruginosa algae were found at high concentrations in some cases. No pesticides were detected. THM and BTEX were below guideline values into recommendations for drinking water. Geosmin was responsible for detected odors. Besides, Copaene, another volatile metabolite, which on account in its structure could be considered a Geosmin precursor, was also detected. In Argentina, this was the first report of Cyanobacteria presence in drinking water.

En abril de 2000, en Bahía Blanca (Argentina), se detectaron alteraciones en el agua de bebida, turbidez y olor semejante a "Gamexane". Esto coincidió con la aparición de problemas dérmicos y respiratorios en la población. Para determinar el origen del problema, se analizaron muestras, tomadas en el embalse Paso de las Piedras, en las plantas potabilizadoras y en varios domicilios particulares. En el embalse, la densidad celular del fitoplancton, fluctuó entre 49440 y 84832 cél/ml-1, muy por encima de los valores de referencia. Anabaena circinalis en altas concentraciones fue la especie dominante (48320 - 84032 células por mililitro). Se cuantificó la eficiencia de las plantas de tratamiento (algas-entrada/algas-salida). Anabaena circinalis y Microcystis aeruginosa fueron encontradas a la entrada y a la salida de las plantas, indicando una baja eficiencia de las plantas de tratamiento. Para el agua domiciliaria, se realizaron estudios fitoplanctonicos; análisis bacteriológicos; presencia de plaguicidas, trihalometanos y BTEX. Si bien, bacteriológicamente el agua era potable, se encontraron Cyanobacteria en algunos casos en concentraciones muy altas. La presencia de pesticidas resultó negativa. La media de los THM y de los valores de BTEX resultaron menores a valores guías para agua potable. Se detectó la presencia de Geosmina, resultando el responsable del fuerte olor. Asimismo, se halló otro metabolito volátil, Copaene, que por su estructura pudiera llegar a considerarse precursor de la Geosmina. Este es el primer reporte de la presencia de cianobacterias en aguas de consumo humano en Argentina.