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A scanner-based microplate platform for high-throughput oxytetracycline detection using diazotization-coupling chemistry with greenness evaluation
(2026-06-01)
Puttasa, Papawarin
;
Ngernpimai, Sawinee
;
Tippayawat, Patcharaporn
;
Park, Myoung‑Hwan
;
Teerasong, Saowapak
Routine oxytetracycline (OTC) monitoring of aquaculture effluents, water sources, and pharmaceutical products is essential for environmental management and regulatory compliance. However, existing analytical methods remain largely instrumental and low-throughput. Herein, a microvolume, high-throughput, scanner-based colorimetric microplate platform is developed for OTC determination using a sensitive and selective diazotization-coupling chemistry with RGB digital image analysis. Sulfanilamide is diazotized with nitrite to generate reactive diazonium species, which subsequently couple with OTC to form a strongly colored yellow-orange azo dye. Only microliter-scale volumes of reagents and samples are required in each well, and the reaction is completed within a few minutes, enabling simultaneous analysis of up to 96 samples. The microplate is imaged with a flatbed scanner, and color responses are quantified using ImageJ software. A linear calibration range of 1 − 40 μM was obtained with a detection limit of 0.33 μM. The platform was successfully applied to OTC determination in aquaculture effluents, tap water, drinking water, and pharmaceutical samples. It exhibited high selectivity against other antibiotics with no observable cross-reactivity. The green analytical procedure index (GAPI) confirmed the method’s environmental friendliness. Overall, the scanner-based microplate platform offers a simple, low-cost, portable, and high-throughput alternative to conventional OTC assays.
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Solid shrimp waste derived nanoporous carbon as an alternative bio-sorbent for oxytetracycline removal from aquaculture wastewater
(2024-06-15)
Kaewtrakulchai, Napat
;
Samattakarn, Nippit
;
Chanpee, Sirayu
;
Assawasaengrat, Pornsawan
;
Manatura, Kanit
Recently, it has been critical to effectively remove oxytetracycline (OTC) from aquaculture wastewater before releasing into the environment. The adsorption process is recognized as an efficient pathway for removing OTC since it is a simple, stable, and cost-effective method. This study aims to develop nanoporous carbon entirely from shrimp waste (SW) via hydrothermal carbonization assisted with KOH activation. Existing KOH significantly increases the porosity of SW nanoporous carbon. The optimal SW porous carbon was obtained using 5 wt%KOH for activation, which had the largest surface area of 679.51 m2/g with the total pore volume of 0.458 cm3/g. Moreover, the SW porous carbon with the highest porosity was selected for the OTC adsorption. The Langmuir isotherm model and the pseudo-second-order kinetic model match the experimental data, implying that the adsorption mechanism is mono-layered adsorption due to micropores by chemisorption interaction. The adsorption capacity significantly improved by increasing the dosage of SW nanoporous carbon. The SW nanoporous carbon adsorption for OTC is primarily regulated by pore filling affected by hydrogen bonding, and π-π* interaction also plays a significant role. The SW nanoporous carbon showed an efficient OTC adsorption after 5 regeneration cycles. This work demonstrates biomass waste recycling and emphasizes the potential of aquatic food processing waste-derived nanoporous carbon for antibiotic adsorption.
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Diversity and characterization of oxytetracycline-resistant bacteria associated with non-native species, white-leg shrimp (Litopenaeus vannamei), and native species, black tiger shrimp (Penaeus monodon), intensively cultured in Thailand
(2011-01-01)
Yano, Y.
;
Hamano, K.
;
Satomi, M.
;
Tsutsui, I.
;
Aue-Umneoy, D.
Aims: This study aimed at surveying prevalence of oxytetracycline (OTC)-resistant bacteria in the white-leg shrimp Litopenaeus vannamei, and the black tiger shrimp Penaeus monodon, intensively cultured in Thailand. We investigated the phylogenetic diversity of the bacterial isolates, as well as the minimum inhibitory concentration (MIC) of OTC, the occurrence of major OTC-resistant genes and multiple-antibiotic resistance in the isolates. Methods and Results: Shrimps were collected from culture ponds, and the homogenates of whole bodies were plated on tryptic soy agar supplemented with or without OTC. Percentages of OTC-resistant bacteria were 0·3-52·1% in white-leg samples and 0·008-22·3% in black tiger samples. Analyses of 16S rDNA sequences indicated that most OTC-resistant isolates were closely related to Aeromonas spp. and Lactococcus garvieae. MICs of OTC were 4-128μgml-1 in the OTC-resistant aeromonads and 128-256μgml-1 in OTC-resistant L. garvieae. OTC resistance was found to be conferred by the genes tet(A), tet(C), tet(D), tet(E), tet(M) and tet(S), detected either singly or in pairs. No resistance to ceftazidime, imipenem or chloramphenicol was observed in any isolate. Conclusions: Both species of shrimp are associated with OTC-resistant bacteria, occasionally at high densities exceeding 106cfug-1. The associated bacteria, predominantly Lactococcus and Aeromonas genera, are potential pathogens and are reservoirs of a variety of OTC-resistant genes. Significance and Impact of the Study: Cultured shrimps can be vehicle to carry OTC-resistant bacteria to domestic and foreign consumers via the food chain. Very low populations of OTC-resistant bacteria observed in the several ponds suggest that levels of the resistant bacteria are artificially high and should be reduced in farmed shrimps. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

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