No abstract

The fate of malachite green and its main metabolite leucomalachite green during thermal treatment was examined in seafood (brook trout and white shrimp) using non-target analysis. Samples were extracted using QuEChERS and analyzed using liquid chromatography coupled with quadruple time of flight mass spectrometry. Malachite green levels were reduced in meat during boiling (∼40%), microwaving (64%), and canning (96%). Only microwaving was successful in significantly decreasing leucomalachite green levels in brook trout. The reduction percentages of the two target analytes were not significantly different in shrimp (mean fat content = 0.8 ± 0.3%) and in brook trout (mean fat content = 3.5 ± 1.7%), suggesting that a higher fat content may not affect the reduction of the more lipophilic leucomalachite green in these two matrices. Three transformation products were tentatively identified in the cooked tissues, resulting from the cleavage of the conjugated structure or through demethylation. Further research is needed to determine possible adverse health effects. The findings of this study show how non-target analysis can complement targeted methodologies in identifying and evaluating risks to human health.

Surface waters, especially those receiving wastewater flows, can disseminate antimicrobial resistant bacteria (ARB), antimicrobial resistance genes (ARG), and antibiotics. In the Scioto River of central Ohio, United States, we evaluated fishes as potential sentinels of ARB and antimicrobial contamination and investigated the influence of antimicrobial exposure on the fish intestinal resistome. Seventy-seven fish were collected from river reaches receiving inputs from two wastewater treatment plants that serve the greater Columbus Metropolitan Area. Fish were screened for the presence of cephalosporin-resistant (CeRO) and carbapenem-resistant (CRO) organisms, epidemic carbapenemase genes, and antibiotic drugs and metabolites using culture methods, droplet digital PCR, and ultra-high performance liquid chromatography tandem mass spectroscopy (UHPLC-MS/MS). Nearly 21% of fish harbored a CeRO in their resistome, with 19.4% exhibiting bacteria expressing an AmpC genotype encoded by blaCMY, and 7.7% with bacteria expressing an extended-spectrum β-lactamase phenotype encoded by blaCTX-M. blaKPC and blaNDM were present in 87.7% (57/65) and 80.4% (37/46) of the intestinal samples at an average abundance of 104 copies. Three antibiotics–lincomycin (19.5%), azithromycin (31.2%) and sulfamethoxazole (3.9%)–were found in hepatic samples at average concentrations between 25–31 ng/g. Fish harboring blaCTX-M and those exposed to azithromycin were at greater odds of being downstream of a wastewater treatment plant. Fish that bioconcentrated antibiotics in their liver were not at greater odds of harboring CeRO, CRO, or epidemic carbapenemase gene copies in their resistome. Our findings confirm that fishes can be effective bioindicators of surface waters contaminated with ARB, ARG, and antibiotics. Moreover, our findings highlight the varying importance of different mechanisms that facilitate establishment of ARB in aquatic ecosystems.

No abstract

Oxytetracycline is an antibiotic authorized for use in aquaculture and is often detected in seafood products, especially shrimp. Previous studies investigating the fate of oxytetracycline in shrimp tissues after cooking were limited to quantification of parent compound residues, and did not describe any potential transformation products formed. Hence, the main objective of this study was to apply a non-target analysis workflow to study the fate of oxytetracycline in shrimp muscle. Furthermore, “water” and “spiked” models were evaluated if they were suitable to track the transformation of OTC in incurred muscle, and determine if the matrix plays a role in the transformation pathway. First, four different extraction methods were compared for the determination of oxytetracycline in muscle. Secondly, raw and cooked samples were then extracted using the suitable method (acidified water/methanol/acetonitrile with clean-up of samples achieved using freezing) and were analyzed by HPLC-QTOF-MS. Oxytetracycline levels were reduced by 75 % and 87% in muscle and water, respectively. Identification of thermal transformation products was limited to formula generation, but results showed that different compounds were identified in spiked and incurred muscle.