Evaluation of precopulatory pairing behaviour and male fertility in a marine amphipod exposed to plastic additives.

Plastics contain a mixture of chemical additives that can leach into the environment and potentially cause harmful effects on reproduction and the endocrine system. Two of these chemicals, N-butyl benzenesulfonamide (NBBS) and triphenyl phosphate (TPHP), are among the top 30 organic chemicals detected in surface and groundwater and are currently placed on international watchlist for evaluation. Although bans have been placed on legacy pollutants such as diethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP), their persistence remains a concern. This study aimed to examine the impact of plastic additives, including NBBS, TPHP, DBP, and DEHP, on the reproductive behaviour and male fertility of the marine amphipod Echinogammarus marinus. Twenty precopulatory pairs of E. marinus were exposed to varying concentrations of the four test chemicals to assess their pairing behaviour. A high-throughput methodology was developed and optimised to record the contact time and re-pair time within 15 min and additional point observations for 96 h. The study found that low levels of NBBS, TPHP, and DEHP prolonged the contact and re-pairing time of amphipods and the proportion of pairs reduced drastically with re-pairing success ranging from 75% to 100% in the control group and 0%-85% in the exposed groups at 96 h. Sperm count declined by 40% and 60% in the 50 µg/l and 500 µg/l DBP groups, respectively, whereas TPHP resulted in significantly lower sperms in 50 µg/l exposed group. Animals exposed to NBBS and DEHP showed high interindividual variability in all exposed groups. Overall, this study provides evidence that plastic additives can disrupt the reproductive mechanisms and sperm counts of amphipods at environmentally relevant concentrations. Our research also demonstrated the usefulness of the precopulatory pairing mechanism as a sensitive endpoint in ecotoxicity assessments to proactively mitigate population-level effects in the aquatic environment.

Cross preferences and genetic diversity of Psidium interspecific hybrids through morphoagronomic traits and resistance to Meloidogyne enterolobii.

The introgression of M. enterolobii resistance-related genes in guava breeding programs can be compromised by incompatibility among Psidium species. This study aimed to evaluate the female parent preference and genetic diversity of Psidium interspecific hybrids using morphoagronomic traits and resistance to M. enterolobii. There were evaluated cross successes and germination from crosses between accesses of P. cattleyanum, P. guineense and P. guajava and the genetic diversity by Ward-MLM method of hybrids according to descriptors developed for the genus. Crosses were more successful when P. cattleyanum was the female parent. Germination was more successful in crosses involving P. cattleyanum and P. guajava. Four groups were formed. The group IV clustered the most resistant genotypes, composed by genotypes of P. cattleyanum x P. guineense, while the group II was the most susceptible. The groups I and III grouped some genotypes of P. cattleyanum x P. guajava with low levels of susceptibility. There are preferences of female parent species among crosses. Some individuals of groups I and III can be used as source of resistance genes for the breeding program, due the presence of favorable alleles inherited from guava parent. The high susceptibility leads to reduction in root development.

Occurrence and distribution of legacy and emerging pollutants including plastic debris in Antarctica: Sources, distribution and impact on marine biodiversity.

Since the first explorers reached Antarctica, their activities have quickly impacted both land and sea and thus, together with the long-range transport, hazardous chemicals began to accumulate. It is commonly recognized that anthropogenic pollution in Antarctica can originate from either global or local sources. Heavy metals, organohalogenated compounds, hydrocarbons, and (more recently) plastic, have been found in Antarctic biota, soil sediments, seawater, air, snow and sea-ice. Studies in such remote areas are challenging and expensive, and the complexity of potential interactions occurring in such extreme climate conditions (i.e., low temperature) makes any accurate prediction on potential impacts difficult. The present review aims to summarize the current state of knowledge on occurrence and distribution of legacy and emerging pollutants in Antarctica, such as plastic, from either global or local sources. Future actions to monitor and mitigate any potential impact on Antarctic biodiversity are discussed.

Meloidogyne enterolobii-induced Changes in Guava Root Exudates Are Associated With Root Rotting Caused by Neocosmospora falciformis.

Despite the worldwide importance of disease complexes involving root-feeding nematodes and soilborne fungi, there have been few in-depth studies on how these organisms interact at the molecular level. Previous studies of guava decline have shown that root exudates from Meloidogyne enterolobii-parasitized guava plants (NP plants), but not from nematode-free plants (NF plants), enable the fungus Neocosmospora falciformis to rot guava roots, leading to plant death. To further characterize this interaction, NP and NF root exudates were lyophilized; extracted with distinct solvents; quantified regarding amino acids, soluble carbohydrates, sucrose, phenols, and alkaloids; and submitted to a bioassay to determine their ability to enable N. falciformis to rot the guava seedlings' roots. NP root exudates were richer than NF root exudates in amino acids, carbohydrates, and sucrose. Only the fractions NP-03 and NP-04 enabled fungal root rotting. NP-03 was then sequentially fractionated through chromatographic silica columns. At each step, the main fractions were reassessed in bioassay. The final fraction that enabled fungal root rotting was submitted to analysis using high performance liquid chromatography, nuclear magnetic resonance, mass spectrometry, energy-dispersive X-ray fluorescence, and computational calculations, leading to the identification of 1,5-dinitrobiuret as the predominant substance. In conclusion, parasitism by M. enterolobii causes an enrichment of guava root exudates that likely favors microorganisms capable of producing 1,5-dinitrobiuret in the rhizosphere. The accumulation of biuret, a known phytotoxic substance, possibly hampers root physiology and the innate immunity of guava to N. falciformis.

The amphipod Parhyale hawaiensis as a promising model in ecotoxicology.

Near-shore marine/estuarine environments play an important role in the functioning of the marine ecosystem and are extremely vulnerable to the presence of chemical pollution. The ability to investigate the effects of pollution is limited by a lack of model organisms for which sufficient ecotoxicological information is available, and this is particularly true for tropical regions. The circumtropical marine amphipod Parhyale hawaiensis has become an important model organism in various disciplines, and here we summarize the scientific literature regarding the emergence of this model within ecotoxicology. P. hawaiensis is easily cultured in the laboratory and standardized ecotoxicity protocols have been developed and refined (e.g., miniaturized), and effects of toxicants on acute toxicity (Cd, Cu, Zn, Ag, ammonia, dyes, pesticides, environmental samples), genotoxicity as comet assay/micronuclei, and gene expression (Ag ion and Ag nanoparticles) and regeneration (pesticides) have been published. Methods for determination of internal concentrations of metals (Cu and Ag) and organic substances (synthetic dye) in hemolymph were successfully developed providing sources for the establishment of toxicokinetics models in aquatic amphipods. Protocols to evaluate reproduction and growth, for testing immune responses and DNA damage in germ cells are under way. The sensitivity of P. hawaiensis, measured as 50% lethal concentration (LC50), is in the same range as other amphipods. The combination of feasibility to culture P. hawaiensis in laboratory, the recent protocols for ecotoxicity evaluation and the rapidly expanding knowledge on its biology make it especially attractive as a model organism and promising tool for risk assessment evaluations in tropical environments.

Genotoxic effects of silver nanoparticles on a tropical marine amphipod via feeding exposure.

Silver nanoparticles (AgNP) are widely used in several applications including as antifouling agents; therefore, they can end up in estuarine and marine environments. These nanoparticles tend to aggregate and to deposit in the sediment, where many organisms feed and reproduce. Parhyale hawaiensis is an epibenthic amphipod globally distributed in tropical zones, and has been considered a potential model for ecotoxicology. The aim of this study was to evaluate genotoxic effects of AgNP and Ag salt via feeding, as P. hawaiensis lives in the sediment where nanoparticles tend to accumulate. Organisms were cultivated in the laboratory, and adults were exposed to food containing both AgNP and Ag salt. We collected hemolymph after different times of exposure, and analysed the hemocytes for nuclear abnormalities (including micronuclei) and DNA damage using the standard alkaline comet assay. Conditions of both assays were developed/optimized to allow their successful application in marine invertebrates. Increased frequencies of micronuclei, nuclear buds and total abnormalities were detected in relation to concentration and time in organisms exposed to AgNP and Ag salt in comparison to the controls. No DNA damage was detected when the alkaline comet assay was applied. After 5 days of exposure, we observed higher micronuclei frequencies in Ag salt treatment in comparison with AgNP. After 13 days, micronuclei frequencies were similar for both silver forms. We believe that the Ag, in its ion form, is causing the mutagenic effect; therefore, more time would be needed for the release of the ion from AgNP, explaining the delayed mutagenic effect.

Genetic structuring of segregating populations of Psidium spp resistant to the southern root-knot nematode by Bayesian approach as basis for the guava breeding program.

There are no guava cultivars resistant to the Meloidogyne enterolobii; for this reason, genetic breeding has been performed by introgressing genes into the current cultivars through interspecific hybridization. We used 33 microsatellite markers for the genetic-molecular characterization of segregating populations of Psidium resistant to M. enterolobii, aiming at selection within and between populations for generation advancement in the guava breeding program. The average number of alleles per locus ranged from 1.60 to 2.09. Populations 1 (P. guineense × P. cattleyanum) and 5 (P. guajava × P. cattleyanum) obtained the greatest genetic diversity, which can be confirmed by the higher observed-heterozygosity values (0.422 and 0.312, respectively). Bayesian analysis showed that the populations were subdivided into three groups, agreeing with the number of groups observed by Nei's genetic distance. The population obtained from the P. guineense × P. cattleyanum cross differed from the others with a clear structuring, whereas the P. guajava × P. cattleyanum and P. cattleyanum × P. guineense populations were the most similar between each other. The SSR markers were efficient in discriminating the populations, and individual 80 may be employed in future crosses with guava, allowing generation advancement in the guava breeding program aimed at resistance to M. enterolobii.

Genotoxicity, potential cytotoxicity and cell uptake of titanium dioxide nanoparticles in the marine fish Trachinotus carolinus (Linnaeus, 1766).

Nanoparticles have physicochemical characteristics that make them useful in areas such as science, technology, medicine and in products of everyday use. Recently the manufacture and variety of these products has grown rapidly, raising concerns about their impact on human health and the environment. Adverse effects of exposure to nanoparticles have been reported for both terrestrial and aquatic organisms, but the toxic effects of the substances on marine organisms remain poorly understood. The main aim of this study was to evaluate the genotoxicity of TiO2-NP in the marine fish Trachinotus carolinus, through cytogenotoxic methods. The fish received two different doses of 1.5 µg and 3.0 µg-TiO2-NP g(-1) by intraperitoneal injection. Blood samples were collected to analyze erythrocyte viability using the Trypan Blue exclusion test, comet assay (pH>13), micronucleus (MN) and other erythrocyte nuclear abnormalities (ENA) 24, 48 and 72 h after injection. The possible cell uptake of TiO2-NP in fish injected with the higher dose was investigated after 72 h using transmission electron microscopy (TEM). The results showed that TiO2-NP is genotoxic and potentially cytotoxic for this species, causing DNA damage, inducing the formation of MN and other ENA, and decreasing erythrocyte viability. TEM examination revealed that cell uptake of TiO2-NP was mainly in the kidney, liver, gills and to a lesser degree in muscle. To the extent of the authors' knowledge, this is the first in vivo study of genotoxicity and other effects of TiO2-NP in a marine fish.

Assessment of trophic transfer of benzo(a)pyrene genotoxicity from the post-larval pink shrimp F. brasiliensis to the juvenile Florida pompano T. carolinus.

In the present study, the polycyclic aromatic hydrocarbon (PAH) genotoxicity was investigated in a one-step predator-prey relationship with the trophic-related marine species. Florida pompanos were fed for 5 and 10 days with pink shrimp post larvae previously exposed to benzo(a)pyrene (BaP) concentrations. Parent BaP body burden was measured in samples of Farfantepenaeus brasiliensis. BaP metabolites were determined in bile samples of Trachinotus carolinus and DNA damage was assessed through the comet and erythrocyte nuclear abnormalities (ENAs) assays in fish erythrocytes. BaP body burden increased significantly with the PAH concentration in pink shrimp PLs as well as the fish bile BaP metabolites. Both, comet and ENAs assays indicated significant increase on erythrocyte DNA damage of Florida pompanos fed with BaP-exposed pink shrimp on both feeding periods. The trophic route of BaP genotoxicity is discussed as well as the PAH biotransformation as the inducing mechanism for the DNA damages observed.

EROD activity and genotoxicity in the seabob shrimp Xiphopenaeus kroyeri exposed to benzo[a]pyrene (BaP) concentrations.

Seabob shrimp Xiphopenaeus kroyeri is a marine species that lives in shallow waters of coastal environments, often impacted by polycyclic aromatic hydrocarbons (PAH) pollution. In the present study, seabob shrimp were exposed for 96 h to benzo[a]pyrene (BaP) at the nominal concentrations of 100, 200, 400 and 800 microg-L(-1). Animals of the control groups were exposed either to clean water or to the BaP-carrier (DMSO). At the end of the exposures, muscle tissues were sampled for BaP uptake assessment and hepatopancreas and hemolymph for EROD enzyme activity and hemocytes DNA damage, respectively. EROD activity and DNA damage increased significantly as a function of BaP exposure concentrations. Significant correlations between BaP uptake and both EROD activity and DNA damage suggest that they can be used as suitable tools for integrated levels of study on the biomarkers of PAH exposure.

Effects of anionic surfactant and salinity on the bioenergetics of juveniles of Centropomus parallelus (Poey).

The acute and sublethal chronic effects of sodium dodecyl sulphate (SDS) on the survival, metabolism, and growth of juveniles of Centropomus parallelus were investigated at three different salinities. Results of 96 h LC50 test showed that juveniles of C. parallelus were very sensitive to SDS in comparison to other species investigated. For each group of exposure to nominal concentrations of SDS (0.10 and 0.25 mg/L) and the control group (0.0 mg/L), at the different salinities (5, 20, and 30) there were significant differences in the specific growth rate, oxygen consumption, and ammonia excretion rates, O:N atomic ratio at the different exposure periods (15 and 30 days). There were also interactions between factors for the parameters investigated. The present results show a pronounced effect of SDS, mainly at the highest concentration and salinity, as well as after a long time of exposure.

Baseline mercury and zinc concentrations in terrestrial and coastal organisms of Admiralty Bay, Antarctica.

This paper provides the first quantitative information on mercury in soil, coastal sediment, and in characteristic organisms of terrestrial and shallow coastal marine ecosystems from Admiralty Bay (King George Island, Antarctica). As expected for a remote area, mercury content is low in abiotic components of the ecosystem, and probably similar to natural levels. Mercury also occurs in very low concentrations in the vegetation, invertebrates and fish. These low mercury levels may be due to sulphide formation in reducing sediments of this environment. Higher concentrations of mercury occurred in bird feathers and mammal hair, indicating biomagnification. This was not found for Zinc. These results may be useful as a reference background to detect future inputs of trace elements in this remote area of the earth. Terrestrial vegetation and bird feathers are suggested as target regional biomonitors.