Environmental concerns about the massive use of disinfectants during COVID-19 pandemic: an overview on aquatic toxicity

Many public health measures to mitigate the spread of SARS-CoV-2 were adopted worldwide, and particularly to the environmental measure of regular cleaning and disinfection of surfaces, the increased use of disinfectant products raises environmental concerns. Quaternary ammonium compounds (QACs), povidone-iodine (PVP-I), chloroxylenol (PCMX) and chlorhexidine (CHX) are the active ingredients of most disinfectant products due to their effectiveness against various microbiological agents. Although presenting antimicrobial efficacy, these biocides have been associated with impacts on aquatic life. For instance, QACs can induce toxicity to Aliivibrio fischeri and fish (different species). Gill and liver damages are verified in Cyprinus carpio after exposure to PVP-I. CHX induces toxic effects on algae, crustaceans, and fish embryos. PCMX can induce genotoxicity to rainbow trout and malformations on zebrafish embryos, as well as it can reduce the reproduction rate of Caenorhabditis elegans. Thus, the potential to cause negative consequences on human and environmental health has resulted in activities from the U.S. and European agencies to favor the use of safer and greener disinfectant products during the COVID-19 pandemic. This review article summarizes the main findings on the impacts of disinfectants (the most used) on aquatic life. This information may help prioritize disinfectants with lower impacts on the aquatic environment for daily use, and especially for high-frequency use as verified in the COVID-19 pandemic. Furthermore, this review may help identify knowledge gaps on the aquatic hazard of disinfectants, which may drive future studies on this matter and, lastly, contribute to the development of sustainable products.

Earthworm coelomocytes as a soil health assessment tool

Different pollutants can disrupt earthworm coelomocytes integrity and functions, and their responses can be applied as biomarkers of sublethal contaminant exposure. In this context, the aim of this study was to develop an in vitro protocol for coelomocyte extraction, maintenance and analysis with regard to soil health status and chemical toxicity profile assessments. The extrusion technique was first tested comparing previously depurated (purged stomach content) and non-depurated and resampled earthworms. After testing, earthworms were exposed to different 2,4D and chloroacetamide concentrations for methodology validation. The values of viability were not affected by food restriction since no statistical difference was observed between non-depurated (sample A) and depurated (sample B) organisms. Regarding to cell density, a significant (p<0;05) reduction of 22% was observed between non-depurated and depurated organisms, indicating that food restriction may affect cell density. However, the non-depurated resampling did not show a significant reduction, indicating that this assessment may not be affect by resampling of the same organism. For both chemical compounds, no change in cell viability was observed at all assessed concentrations and exposure times. However, for cell density, a mainly time-dependent effect was observed for organisms exposed to chloroacetamide, and concentration-dependent effect for organisms exposed to 2,4D. The proportion of immune system cells was altered, mainly after 24 h, with the increasing of granular amoebocytes proportion. The difference in the proportion of granular amoebocytes in earthworms exposed to 2,4D can be explained by the existence of recognition and elimination mechanisms for this chemical substance. Thus, assessments of pollutant responses with in vitro coelomocytes seem to be a powerful tool for ecotoxicological studies.

Use of LC50 in aquatic regulatory toxicology-Disharmony in global harmonization of hazard classification of chemicals

In regulatory aquatic toxicology, acute toxicity studies with chemicals are conducted with a species of fish, crustacea, and or alga. The LC50/EC50 obtained from these studies is used for the hazard classification and labeling of the chemicals. The methods like probit or logit analysis and Litchfield and Wilcoxon method are prescribed in the OECD guidelines to determine the LC50. In the present study, LC50s were calculated using probit analysis, Litchfield & Wilcoxon method, and also using the method by Trevan (the inventor of median lethal dose) using three sets of concentration-mortality data of fish acute toxicity tests. The slopes of the concentration-mortality curves, fiducial limits (95% confidence interval) of LC50s, and ‘mode’ of the concentration-mortality curves were compared. Though the methods used in the study resulted in more or less similar LC50s, the LC10 and LC90, slopes and ‘mode’ differed considerably, indicating that LC50 does not reveal the exact toxicity profile of a chemical. The LC50 calculated using Finney’s probit analysis provides better information on the toxicity profile of a chemical than the LC50calculated by Litchfield & Wilcoxon method. While interpreting LC50, the mortality occurred below 16 % (eg.,LC10) and above 84 % (eg.,LC90), slope and ‘mode’ of the concentration-mortality curve may also be considered. It is worth having a relook at the current practice of hazard classification and labeling of the chemicals based only on LC50 in regulatory aquatic toxicology.

https://ecotoxbrasil.org.br/revista/28/ecotoxicology-and-environmental-contamination/

Glyphosate is an active ingredient used in herbicide formulations worldwide. However, besides glyphosate, these formulations have other components to facilitate glyphosate absorption by plants. These include the surfactants such as polyoxyethylene amine (POEA) present in the Roundup Transorb® (RT) formulation. Glyphosate formulations are potentially more toxic to non-target organisms than the pure active ingredient. In this work, we evaluated the toxicity (72 h) of pure glyphosate and RT for the cyanobacterium Synechococcus elongatus, based on biomass growth and cell viability. The formulation proved more toxic than pure glyphosate for both parameters analysed, with an IC50 (Inhibition concentration mean) based on biomass measured by optical density (750 nm) that was sixty times lower. Cell viability was not as sensitive as the biomass because, of the few cells left in the culture, most were viable. This indicates that there is a variation in tolerance between the cyanobacteria present in the inoculum. Thus, cell viability may underestimate the results of glyphosate and RT toxicity and be useful only in low concentrations of exposure

Histopathological effects of silver nanoparticles in Rhamdia quelen after oral exposure

The studies about silver nanoparticles (AgNP) increased in the last years but few is known about their effects in Brazilian neotropical freshwater fish species. The current study investigated the effects of AgNP on adult silver catfish Rhamdia quelen after subchronic oral exposure. After nanoparticle (NP) size and area characterization fish were administrated with three different doses for 15 days (0.03, 0.3 and 3 µg g-1). The concentration of silver in liver and kidney was measured to evaluate the bioaccumulation and discuss its effects in the target organs. Liver bioaccumulated 15, 1.7 and 0.2 % of administered doses while kidney bioaccumulated 1.33, 0.33 and 0.9 % (respectively for 0.03, 0.3 and 3 µg g-1). The histopathological findings were considered in both organs to evaluate the effects of AgNP, according to Bernet’s Lesion Index (BLI). Also were included the melano-macrophages center (MMC) and new nephrons (NN) counting respectively in liver and posterior kidney. The results revealed morphological injuries as inflammation in both studied organs and vascular congestion and steatosis in liver, in a concentration dependent way. The presence of AgNP in the tissues revealed the bioavailability of the nanoparticle while the damages and morphological disturbs showed the potential risk of exposure in R. quelen, even under environmental relevant concentrations.

Cyanobacteria reduction in the integrated culture of Pacific white shrimp and West India pointed venus in a biofloc system

This study aimed to evaluate the integrated culture of the Pacific white shrimp (Litopenaeus vannamei) and the West Indian pointed venus (Anomalocardia flexuosa) reared in a biofloc system. The performance of both species and the phytoplankton community were evaluated. Shrimp monoculture and three stocking densities of West Indian pointed venus (2.5, 5.0 and 7.5 Kg m-3) were evaluated in 0.05 m3 tanks. First of all, the addition of A. brasiliana did not affect the water quality parameters of the culture. Regarding the phytoplankton community, 13 taxa were found: 5 cyanobacteria, 4 dinoflagellates, 3 diatoms and 1 green algae. Although diatoms were dominant at the beginning of cultivation, in the middle and at the end a cyanobacterial dominance was observed (proven by ANOSIM and nMDS). The taxa that most contributed to the abundance of cyanobacteria were Aphanocapsa sp. and Pseudanabaena sp. Regarding the performance of the shrimp, all integrated treatments present the higher growth performances (final weight, yield and survival) when compared to shrimp monoculture. Moreover, gradual mortalities of A. brasiliana were observed, probably due to the high concentration of cyanobacteria. Thus, it can be observed that the integration of molluscs in shrimp culture can reduce the concentration of harmful algae and improve the zootechnical performance of the shrimp.

Investigating the toxicity of tropical reservoir sediments using the Allium test

Water pollution is a global environmental issue, and aquatic sediments are important compartments that might act as sinks or sources of contaminants. Once in the environment, inorganic contaminants such as metals can cause cytogenotoxic effects that damage genetic material and harm the aquatic community. Biological assays such as the Allium test can be used to investigate potential cytogenotoxicity of contaminated sediments based on the alterations of cell cycle indexes and chromosomal aberration frequencies. Therefore, we aimed to assess the toxicity of sediments from four Brazilian reservoirs using the Allium test. Sediments were sampled and elutriates were prepared in a simulating sediment resuspension in the water column. The Allium test was applied to the elutriates, and the metals copper, chromium, cadmium, lead, zinc, and iron were quantified. The elutriates derived from reservoir sediments were able to reduce the mitotic and anaphase index, increase the prophase and metaphase index, and boost chromosomal aberrations compared to the negative control. The cytogenotoxic effects observed may be linked to the presence of copper, zinc, and iron. Therefore, our results showed that the Allium test was a sensitive tool for warning the occurrence of genotoxic contaminants in sediment elutriates from four Brazilian reservoirs.

Copper affect the multixenobiotic resistance mechanism (MXR) in embryo and larvae of Rhamdia quelen

P-glycoproteins (P-gp) and Multidrug resistance protein (MRP) represent a family of ABC (ATP-binding cassette) transporters responsible for multixenobiotic resistance mechanism (MXR) in aquatic organisms. In the current study the modulation of P-gp and MRP proteins was evaluated in embryo and larvae of Rhamdia quelen fish species exposed to copper. Adult females were exposed by gavage during 60 days to copper (5 mg Cu kg-1) and eggs, embryos, and larvae from exposed and unexposed females were exposed to 30 mg Cu L-1. The activity of ABC transporters was accessed via calcein accumulation assay using the specific inhibitors: Verapamil (P-gp) and MK571 (MRP). P-gp activity was detected in all analyzed stages whereas MRP activity was observed after 36 and 96 hpf. Oocytes from females previously exposed and larvae stages (36 and 96 hpf) accumulated less calcein than no exposed oocytes, showing higher ABC transporters activity. In individuals exposed to copper, a higher inhibitory effect was observed 1 hpf. The modulation of ABC transporter proteins is time dependent throughout the development, and the initial stages are more sensible to copper. These findings highlight the MXR mechanism as a biomarker of pollutant exposure in early stages of development of R. quelen.

Dichlorvos and Paraquat induced spatial avoidance response: A more realistic determinant of population decline of Oreochromis niloticus

The present study evaluated the ability of Dichlorvos and Paraquat to provoke avoidance response in fingerlings of Nile tilapia Oreochromis niloticus and estimate the population immediate decline (PID). The non-forced multi-compartmented system used for non-forced assays, were constructed to allow free movement of fishes along six compartments. Fishes (n=3 per compartment/treatment, totaling 18 per system) were exposed to a gradient of Dichlorvos (1.0, 2.0, 4.0, 5.0 and 6.0 mg L-1) and Paraquat (10.0, 30.0, 50.0, 70.0, 100.0 mg L-1) and their distribution were recorded at 20 min interval for a 3-h period. Mortalities recorded in forced exposures were 17% and 0% at lowest concentrations and, 67% and 83% at highest concentrations for Dichlorvos and Paraquat correspondingly. For non-forced exposure, fishes presented a significant (p < 0.005) gradient-dependent spatial avoidance for both pesticides after 3-h. They avoided the lowest concentrations of Dichlorvos and Paraquat (1.0 and 10.0 mg L-1) by 40% and 90% respectively and 100% at the highest concentrations for both pesticides. The PID was driven by avoidance behavior rather than mortality. This result indicates that the dangers of pesticide contamination is not only in their toxicity to organisms, but also, in habitat selection processes by organism resulting in serious environmental turbulence.

Microcystin bioaccumulation in Limnoperna fortunei following Microcystis aeruginosa exposure, analysis of in vivo enzymatic phosphatase, acetylcholinesterase and carboxylesterase effects and in vitro experiments

Toxic cyanobacteria blooms have been reported in freshwater sources worldwide and may lead to aquatic biota toxin accumulation and trophic chain transfer, resulting in ecological and public health concerns. To assess cyanobacteria effects on microcystin uptake and accumulation and on phosphatase, acethylcholinesterase (AChE) and carboxylesterase (CarbE) enzymatic activities, an in vivo experiment was carried out employing the golden mussel Limnoperna fortunei. These mussels were exposed to a Microcystis aeruginosa NPLJ-4 strain (NPLJ-4) for 48 hours at different cell densities. Subsequently, algal cell counts were carried out and enzymatic activities were assayed. All three enzymes (Phosphatase, AChE and CarbE) were inhibited at the end of the exposure experiment. Mussels exposed to higher in vivo M. aeruginosa densities exhibited microcystin uptake and accumulation. In vitro assays were also carried out, exposing soluble L. fortunei enzyme fractions to M. aeruginosa extracts containing microcystin, and phosphatase inhibition was observed, whereas acetylcholinesterase and carboxylesterase were not inhibited. The results indicate that metabolites other than mycrocystin probably caused the observed in vivo esterase inhibitions, requiring further investigations.