Evaluation of methods for reducing epibionts during farming of the mangrove oyster Crassostrea gasar (Adanson, 1757).

Due to the competition for food, space, oxygen and due to their role as diseases vector, epibionts can negatively affect oyster farming. We assessed the efficacy of six methods commonly used for the removal of epibionts from oyster shells during farming. The experiment was conducted at an oyster farm on the Paraná coast - South Brazil. Oysters (Crassostrea gasar) were acclimated for 90 d in the cultivation system and later exposed to cleaning treatments: i) freshwater; ii) hypersaline water; iii) sodium hypochlorite solution; iv) quaternary ammonia solution; v) exposure to air; vi) hydroblasting; and vii) no cleaning procedure (control). After treatment, oysters were kept in the cultivation system for 15 and 30 d - when the total incrustation and mortality were measured. Epibionts from nine phyla were identified. The most abundant were Arthropoda (Crustacea) (62.5%), Mollusca (33.8%) and Annelida (3.1%). Freshwater [15 (n = 2263 epibionts) and 30 days (n = 2822 epibionts)] and hydroblasting [15 (n = 1850 epibionts) and 30 days (n = 2389 epibionts)] treatments were the most efficient to reduce epibionts and caused lower rates of oyster mortality [15 (5.0 and 3.33%, respectively) and 30 days (1.67 and 6.67%, respectively)].

Metagenomic analysis of the bacterial microbiota associated with cultured oysters (Crassostrea sp.) in estuarine environments.

In this work, we identified the bacterial microbiota associated with farmed oystersin estuarine regions of four states in the north eastern region of Brazil. During the drought and rainy seasons, for eight months, twenty oysters were sampled seasonally from seven different marine farms. In the laboratory, DNA extraction, amplification, and sequencing of the 16S rRNA gene were performed to establish the taxonomic units. We identified 106 genera of bacteria belonging to 103 families, 70 orders, 39 classes, and 21 phyla. Out of the total, 40 of the genera represented bacteria potentially pathogenic to humans; of these, nine are known to cause foodborne diseases and six are potentially pathogenic to oysters. The most prevalent genera were Mycoplasma, Propionigenium, Psychrilyobacter, and Arcobacter. The results indicate the need for more systematic monitoring of bacteria of the genus Mycoplasma in oyster farming operations in the Brazilian north eastern region. Currently, Mycoplasma is not one of the microorganisms analysed and monitored by order of Brazilian legislation during the oyster production and/or commercialization process, even though this genus was the most prevalent at all sampling points and presents pathogenic potential both for oysters and for consumers.

Potential of the Biotic Ligand Model (BLM) to Predict Copper Toxicity in the White-Water of the Solimões-Amazon River.

In this study, we evaluated the capacity of the Biotic Ligand Model (BLM) to predict copper toxicity in white-waters of the Solimões-Amazon River. LC50 tests using the species Otocinclus vittatus (Regan, 1904) were performed with Solimões-Amazon river water (100%) at 20%, 40%, 60%, and 80% dilutions. A sevenfold decrease in both dissolved and total Cu toxicity was observed in the experiment conducted with 100% when compared to 20% white-water, indicating that physicochemical characteristics of white-water attenuate Cu toxicity. There was agreement between the observed LC50 and the LC50 predicted by the BLM after the adjustment of critical accumulation concentration (LA50) for O. vittatus. BLM modeling indicated that dissolved organic carbon (DOC) and pH were the most important water parameters influencing Cu toxicity, followed by Ca2+. Our results highlight the first evidence that the BLM presents potential to predict Cu toxicity to aquatic organisms in the white-water of the Solimões-Amazon River.

The concentrations of polycyclic aromatic hydrocarbons in fish: A systematic review.

This study was conducted on the concentrations of PAHs, including benzo[a]pyrene and the sum of four PAHs (benz[a]anthracene, chryseno, benzo[b]fluoranthene, and benzo[a]pyrene), in unsmoked fish tissues. The values were related to the time period, locality (country and region), living habits, eating habits, and types of environments of the fish species. The data obtained has been also compared with the limits established by the United States Environmental Protection Agency (U.S. EPA) and European Union (E.U.) legislation. The data were collected through a PRISMA systematic review (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) from 1982 to 2020. Only documents that presented the PAHs concentration values were selected. China, Nigeria, and Iran had the highest number of studies. The species Coptodon zillii, Siganus punctatus, and Liza abu were the most commonly used bioindicators. Fish inhabiting demersal and pelagic environments showed the highest concentrations of PAHs. The majority of PAH concentrations of the analyzed documents fell within permissible limits for human consumption, representing a total percentage of 79.49 % for benzo[a]pyrene and 82.86 % for the sum of the 4 PAHs. Although some studies identified concentrations of benzo[a]pyrene alone and benzo[a]pyrene, benz[a]anthracene, chryseno, benzo[b]fluoranthene above the legislation limits, in most of them, the concentrations reported in fish tissue were below the limits defined by European Legislation for human consumption.

Aromatic hydrocarbons extracted by headspace and microextraction methods in water-soluble fractions from crude oil, fuels and lubricants.

Two extraction protocols were developed for the determination of mono- and poly-aromatic hydrocarbons in water-soluble fractions from gasoline, diesel, crude, mineral insulating, and lubricant oils. Development of the procedures was based on clean miniaturized strategies, such as headspace extraction and vortex-assisted dispersive liquid micro-extraction, together with quantification by gas chromatography-mass spectrometry. The mono-aromatic hydrocarbons were extracted using the headspace extraction method. The linear range obtained was 10-500 µg L-1, with r2 > 0.99. Based on the parameters of the analytical curves, detection and quantification limits of 2.56-3.20 and 7.76-9.71 µg L-1 were estimated. In addition, the method showed adequate recoveries of 69.4-83.5%, with a satisfactory precision of 4.7-17.1% (n = 5). Micro-extraction was applied for the poly-aromatics and the most favorable variables were sample volume (5.00 mL) in sodium chloride medium (1%, w/v), trichloromethane as extractor solvent (75 µL), acetone as disperser (925 µL) and vortexing for 1 min. Under these conditions, analytical curves of 0.15-4.00 µg L-1 were obtained and limits of determination and quantification were 0.03-0.15 and 0.09-0.46 µg L-1, respectively. Recovery values of 87.6-124.5% and a maximum relative standard deviation of 18.9% (n = 5) verify satisfactory accuracy and precision. This led to the achievement of enrichment factors for poly-aromatic hydrocarbons of 41-89 times. Finally, the methods were employed in samples of water-soluble fractions for the determination of analytes. The values followed the order: gasoline > diesel > crude > lubricant > mineral insulating oil. These results indicate an increase in lighter fractions, followed by poly-aromatics in more refined products.

Response of Oreochromis niloticus (Teleostei: Cichlidae) exposed to a guanitoxin-producing cyanobacterial strain using multiple biomarkers.

Changes in environmental conditions in aquatic ecosystems caused by anthropic actions can modify the composition of primary producers, promoting the excessive proliferation of cyanobacteria. These organisms can form cyanobacterial blooms, which directly affect aquatic life. The present study investigated the mutagenicity of the cyanobacterium Sphaerospermopsis torques-reginae (strain ITEP-024), guanitoxin-producing (natural organophosphate), and sublethal effects on fish in relevant environment concentrations. For this, the Ames test (Salmonella/microsome) was performed as a mutagenic assay for extracts of the ITEP-024 strain. Specimens of Oreochromis niloticus (Teleostei: Cichlidae) were subjected to acute 96 h exposure to different concentrations of aqueous extract of the strain: C = control group; T1 = 31.25 mg/L; T2 = 62.5 mg/L; T3 = 125 mg/L; and T4 = 250 mg/L. Genotoxic, biochemical, osmoregulatory, and physiologic biomarkers were analyzed. Our results showed that the cyanobacterium had a weak mutagenic response for the TA102 strain of Salmonella with and without metabolic activation by S9. Strains TA98 and TA100 were not affected. Fish from treatments T3 and T4 showed changes in oxidative stress (CAT, SOD, and GST enzymes), inhibition of the enzyme acetylcholinesterase activity, micronucleus formation, and osmoregulatory disorders. No guanitoxin accumulation was detected in the different tissues of O. niloticus by LC-MS/MS. Our results showed unprecedented mutagenicity data of the guanitoxin-producing cyanobacteria by the Ames test and biochemical, osmoregulatory, and genotoxic disorders in fish, providing efficient aquatic contamination biomarkers. Despite the great concern related to the presence of guanitoxin in blooms in freshwater ecosystems, its concentration is not yet regulated, and thus there is no monitoring agenda in current legislation.

Shell form and enzymatic alterations in Lottia subrugosa (Gastropoda, Lotiidae) transplanted to a contaminated site.

Studies have shown that shell morphology and enzymatic activities in mollusks are affected by contaminants exposure. However, the correlation between enzymatic activities and the biomineralization process are not fully understood. The present study used a transplant bioassay and field sampling to evaluate shell measurements and the activities of carbonic anhydrase, Ca2+-ATPase, and Mg2+-ATPase in Lottia subrugosa sampled in Brazilian sites under different contamination levels. Results showed that, in general, shells from the reference site (Palmas) were more rounded than the ones from the contaminated site (Balsa). Effects in enzymatic activities in specimens from transplant bioassay were attributed to the known high contaminant levels present at Balsa. While the lack of enzymatic activity alterations during field sampling was attributed to physiological adaptation to contaminants exposure. Enzymatic activities were not correlated to shell biometric parameters in field sampling, indicating that these enzymes were not related to shell alterations detected in the present study.