Roberto Rico-Martínez

Contribution of Water-Soluble and Insoluble Components and Their Hydrophobic/Hydrophilic Subfractions to the Reactive Oxygen Species-Generating Potential of Fine Ambient Aerosols

Relative contributions of water- and methanol-soluble compounds and their hydrophobic/hydrophilic subfractions to the ROS (reactive oxygen species)-generating potential of ambient fine aerosols (D(p) < 2.5 μm) are assessed. ROS-generating (or oxidative) potential of the particulate matter (PM) was measured by the dithiothreitol (DTT) assay. Particles were collected on quartz filters (N = 8) at an urban site near central Atlanta during January-February 2012 using a PM(2.5) high-volume sampler. Filter punches were extracted separately in both water and methanol. Hydrophobic and hydrophilic fractions were then subsequently segregated via a C-18 solid phase extraction column. The DTT assay response was significantly higher for the methanol extract, and for both extracts a substantial fraction of PM oxidative potential was associated with the hydrophobic compounds as evident from a substantial attenuation in DTT response after passing PM extracts through the C-18 column (64% for water and 83% for methanol extract; both median values). The DTT activities of water and methanol extracts were correlated with the water-soluble (R = 0.86) and water-insoluble organic carbon (R = 0.94) contents of the PM, respectively. Brown carbon (BrC), which predominantly represents the hydrophobic organic fraction (referred to as humic-like substances, HULIS), was also correlated with DTT activity in both the water (R = 0.78) and methanol extracts (R = 0.83). Oxidative potential was not correlated with any metals measured in the extracts. These findings suggest that the hydrophobic components of both water-soluble and insoluble organic aerosols substantially contribute to the oxidative properties of ambient PM. Further investigation of these hydrophobic organic compounds could help identify sources of a significant fraction of ambient aerosol toxicity.

Identification of a sex pheromone from a rotifer

The first isolation and characterization of a sex pheromone from a zooplankter is reported. A 29 kdalton glycoprotein on the surface of females of the marine rotifer Brachionus plicatilis acts as a contact-mating pheromone. This glycoprotein (gp29) is glycosylated with oligosaccharides containing Nacetylglucosamine, mannose, and fucose residues, and these oligosaccharides are necessary for male recognition of females. Males detect this signal by contact chemoreception with receptors located in their corona. Binding of purified glycoprotein to male receptors reduces mating attempts by 93%. An antibody to the glycoprotein binds to females, reducing male mating attempts by 86%. When purified gp29 is bound to sepharose beads, it is sufficient to elicit male mating behavior. This glycoprotein is likely to play a key role in the evolution and maintenance of reproductive isolation in rotifers.

Effect of temperature and food concentration in two species of littoral rotifers

We performed life-table experiments with two species of the littoral rotifers Lecane luna (O.F. Muller, 1776) and Lecane quadridentata (Ehrenberg, 1832). Three different temperatures (20, 25 and 30°C) and food concentrations of Nannochloris oculata (1×107, 5×106, and 1×106 cells ml-1) were investigated. We found important differences between both species in all the treatments regarding offspring sizes, hatching percentages, life span and reproductive rates. Our data on hatching percentages of asexual eggs suggested that the optimal temperature for both species is in the 20–25°C range. On the other hand, reproductive data placed the optimal temperature near 25°C. This information can be used to develop aquatic toxicology tests with littoral species.

Acute toxicity tests on three species of the genus Lecane (Rotifera: Monogononta)

Three rotifer species, Lecane hamata L. luna, and L. quadridentata, were submitted to acute toxicity tests to compare their susceptibility to 11 toxicants. In acute tests with 48-h exposure of neonates of less than 24 h old, copper was most toxic with LC50 values in the range of 0.06–0.33 mg 1−1, while acetone was the least toxic with LC50 values in the range of 5000–7000 mg 1−1. Differences in LC50 value of up to 22-fold were found in the susceptibility to lead between the three species. These data indicate large differences in toxicity among members of the same genus, and point out that it is necessary to submit several species to toxicity tests in order to assess the potential effects of toxicants to rotifers. The commonly used Brachionus calyciflorus cannot be considered representative of all freshwater rotifers in this respect.

Mating behavior and mate recognition pheromone blocking of male receptors in Brachionus plicatilis Müller (Rotifera)

Copulatory behavior of three S and three L type B. plicatilis strains from different geographic areas was analyzed. A 29 KD surface glycoprotein on females, characterized as a Mate Recognition Pheromone (MRP), binds to receptors in the corona of males and blocks mate recognition. Blocking was observed in all S and L strains even though the MRP was isolated from a single L-type strain. Binding was quantified using image analysis and a 20-fold difference was observed among strains. A direct relationship between the male discrimination of females and the intensity of MRP binding to male receptors was found. This relationship might be useful as a tool to examine variation in the mate recognition systems of other rotifer species.

Culture of the rotifer Brachionus calyciflorus Pallas

Abstract Optimum conditions for raising the rotifer Brachionus calyciflorus Pallas were identified. A temperature of 30 °C and a food concentration of 10 7 cells/ml of the alga Chlorella vulgaris were identified as the most suitable conditions for culture of this rotifer. Maximum production was 81 080 rotifers/1 day −1 . The size and growth rate of four different strains of B. calyciflorus were determined. This variation among strains gives this species an advantage over other species in its use as food for larval fish of different sizes because different size strains can be used as food for different sized larval fish.

Male discrimination of female Brachionus plicatilis Müller and Brachionus rotundiformis Tschugunoff (Rotifera)

An analysis of mating behavior of three Brachionus plicatilis O.F. Muller and three Brachionus rotundiformis Tschugunoff strains showed that reproductive isolation exists between strains of these sibling species. An atypical B. rotundiformis strain appears to share some characteristics with both B. plicatilis and B. rotundiformis strains. This strain is able to cross-mate with one B. plicatilis strain, although no cysts were produced. The results of the mating behavior are discussed with regard to recent data on a surface glycoprotein responsible for mate recognition in B. plicatilis. Mating behavior of brachionid rotifers has great potential for maintaining reproductive isolation among species.

Effect of coexposure to DDT and manganese on freshwater invertebrates: Pore water from contaminated rivers and laboratory studies

An environmental survey of several rivers of the southern Huasteca area of Mexico revealed high concentrations of manganese (Mn) and the presence of DDT in the sediments and pore water. Therefore, acute (48-h) toxicities of Mn and DDT were assessed both independently and as a combination on 24-h-old neonates of Daphnia magna Strauss and Lecane quadridentata Ehrenberg. Daphnia magna showed high sensitivity to both toxicants, whereas L. quadridentata was highly resistant to DDT and less susceptible to Mn. For D. magna, the Mn and DDT coexposure was significantly more toxic than any of the singly tested compounds. When D. magna was exposed to sediment pore water, no association was found between the Mn content in the samples and the observed toxicity. Preliminary particle analysis of pore water showed different compounds of Mn, which apparently were not in bioavailable form.

Mating behavior in eight rotifer species: using cross-mating tests to study species boundaries

We characterized the mating behavior of eight freshwater rotifer species from five different families of Monogononts (Asplanchnidae, Brachionidae, Euchlanidae, Lecanidae, and Trichocercidae). Were corded the percentages of male-female encounters resulting in mating attempts and the number of completed copulations. The characterization of mating behavior also included data on: (1) duration of copulation, (2) female and male swimming speed, (3) sites of initial mating attempts, (4) sites of copulation, and (5) sizes of males and females. In the five families investigated, cross-mating tests may be used to assess the ability of males to discriminate conspecifics, establishing limits to gene flow. We also present data on two families, Brachionidae and Lecanidae where we used cross-mating tests to compare discrimination between conspecific females and those of other species. Brachionus calyciflorus males attempted mating with females of the closely related genus Plationus, but did not attempt mating with Keratella Americana females. Males of other three Brachionidae species only attempted mating with nonspecific females. Lecane quadridentata males attempted to mate with females of two different Lecane species besides their own. Percentages of mating attempts and completed copulations were quantified in all crosses. Lower percentages of copulation after a mating attempt were found among brachionids than among non-brachionids. These data demonstrate that male discrimination of females is an effective pre-mating reproductive barrier in several species of Monogonont rotifers. These results, further suggest that investigations of mating behavior are useful for determining species boundaries in rotifers and perhaps other zooplankters.

Comparing toxicity endpoints on Lecane quadridentata (Rotifera: Monogononta) exposed to two anticholinesterases pesticides.

Toxicity tests were performed on the freshwater rotifer Lecane quadridentata exposed to the pesticides carbaryl and methyl parathion (lethal, sublethal, and chronic) to compare the sensitivity between different endpoints: (a) 48‐h mortality; (b) 30‐min in vivo inhibition of esterase activity; (c) 5‐day inhibition of the instantaneous growth rate. The emphasis of this work was to find the most appropriate endpoint to evaluate the toxicity of these pesticides in view of their sensitivity, duration, and ecological relevance. The comparison between the three toxicity tests show that the 5‐day chronic tests have the lowest EC50 (2.22 and 6.6 mg/L), lowest‐observed‐effect concentration (2.5 and 2.5 mg/L), and no‐observed‐effect concentration (1.0 and 1.2 mg/L) values for carbaryl and methyl parathion, respectively. This indicates that the estimate of the instantaneous rate of natural increase r is the most sensitive endpoint regarding the toxicity of these pesticides. This sensitivity might be due to the effect on reducing the growth potential form the first generation on. Lethal and sublethal tests are closely related, suggesting that the immediate effect after inhibition of esterases is death. In general, the sensitivity of L. quadridentata is similar to other species of rotifers exposed to methyl parathion. Therefore, the 5‐day chronic toxicity test with the freshwater rotifer L. quadridentata should be considered a good candidate to evaluate the effect of anticholinesterase pesticides, due to its high sensitivity and ecological relevance.