Characterization of the distribution of antibiotics and their resistance genes in soil profiles of a typical site

doi:10.3969/j.issn.1000-5641.2024.06.007

Abstract

Abstract:

The presence of a variety of environmental contaminants in the soils of steelworks seriously threatens the safety of urban soil and groundwater. Emerging pollutants in deep soils, such as antibiotics and antibiotic resistance genes (ARGs), have not been studied extensively and require timely monitoring and treatment. In this study, the Mingfang Iron and Steel Plant in Handan, Hebei Province, was selected as the research area, and three 4-meter soil columns were collected. The contents of 17 antibiotics in four categories (sulfonamides, fluoroquinolones, macrolides, and tetracyclines) were detected by ultra-performance liquid chromatography-tandem mass spectrometry. The types and abundances of ARGs in the soil at different depths were investigated by metagenomic sequencing. The results showed that the total antibiotic concentrations ranged from 2593.2 to 4279.5 ng/g, among which sulfonamide antibiotics (SAs) were 166.1 to 1103.8 ng/g, fluoroquinolone antibiotics (FQs) were 993.3 to 1330.4 ng/g, macrolide antibiotics (MLs) ranged from 4.0 to 104.8 ng/g, and tetracycline antibiotics (TCs) ranged from 1269.5 to 2077.6 ng/g, with mild to moderate levels of contamination. Soil profile analysis demonstrated that the percentage of SAs were higher in the topsoils (0—150 cm) than in the deep soils (150—400 cm). Contrastingly, MLs accumulated more in deep soils (350—400 cm). The percentages of TCs and FQs were higher in all soil layers and they were the main antibiotic classes detected. The abundance of ARGs corresponding to the four classes of antibiotics decreased with increasing soil depth. The relative abundance of macrolide ARGs was the highest, with the read number of the detected sequences ranging from 0.92 × 10⁵ to 1.44 × 10⁵. The statistical chart of species contributions indicated that Actinobacteria made the greatest contribution to the abundance of ARGs in all four major categories. In conclusion, this study revealed the distribution and migration characteristics of antibiotics in the site soils. Deep soil pollution by ARGs is a matter of concern as it may lead to groundwater contamination and the increased distribution of antibiotic-resistant bacteria.

Key words: steelworks site, soil column, antibiotics, antibiotic resistance genes, metagenomics sequencing

CLC Number:

Mingyin GUAN, Xinran LIU, Min LIU, Jing YANG, Zeying XIE, Qingling ZHANG, Qian LI. Characterization of the distribution of antibiotics and their resistance genes in soil profiles of a typical site[J]. Journal of East China Normal University(Natural Science), 2024, 2024(6): 74-85.

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抗生素种类	英文名称	化学式	分子量
磺胺嘧啶 (SD)	sulfadiazine	C₁₀H₁₀N₄O₂S	250.28
磺胺吡啶 (SP)	sulfapyridine	C₁₁H₁₁N₃O₂S	249.29
磺胺甲恶唑 (SMX)	sulfamethoxazole	C₁₀H₁₁N₃O₃S	253.28
磺胺噻唑 (ST)	sulfathiazole	C₉H₉N₃O₂S₂	255.32
磺胺甲基嘧啶 (SM)	sulfamerazine	C₁₁H₁₂N₄O₂S	264.30
磺胺二甲嘧啶 (SMT)	sulfamethazine	C₁₂H₁₄N₄O₂S	278.33
磺胺喹恶啉 (SQ)	sulfaquinoxaline	C₁₄H₁₂N₄O₂S	300.34
诺氟沙星(NFC)	norfloxacin	C₁₆H₁₈FN₃O₃	319.33
环丙沙星 (CFC)	ciprofloxacin	C₁₇H₁₈FN₃O₃	331.34
恩诺沙星 (EFC)	enrofloxacin	C₁₉H₂₂FN₃O₃	359.40
氧氟沙星 (OFC)	ofloxacin	C₁₈H₂₀FN₃O₄	361.37
四环素 (TC)	tetracycline	C₂₂H₂₄N₂O₈	444.45
土霉素 (OTC)	oxytetracycline	C₂₂H₂₄N₂O₉	460.43
强力霉素 (DXC)	doxycyclinehyclate	C₂₂H₂₄N₂O₈	444.44
金霉素 (CTC)	chlorotetracycline	C₂₂H₂₃ClN₂O₈	478.88
红霉素 (ETM)	erythromycin	C₃₇H₆₇NO₁₃	733.94
罗红霉素 (RTM)	roxithromycin	C₄₁H₇₆N₂O₁₅	837.05

样品	pH值	土壤机械组成/%			总氮/%	总碳/%
样品	pH值	黏土	粉砂	砂	总氮/%	总碳/%
A_0—50	11.7	4.3	93.1	2.7	0.02	2.71
A_100—150	8.0	4.4	85.0	10.6	0.02	1.29
A_200—250	8.0	13.0	85.8	1.2	0.01	0.42
A_350—400	7.6	9.4	89.7	1.0	0.02	0.50
B_0—50	8.1	6.9	92.8	0.4	0.03	1.34
B_100—150	10.9	7.1	92.7	0.2	0.03	1.87
B_200—250	7.2	5.9	91.0	3.2	0.02	0.31
B_350—400	7.6	8.9	87.5	3.6	0.03	0.55
C_0—50	11.3	6.8	75.5	17.7	0.03	4.27
C_100—150	8.5	8.5	91.3	0.2	0.03	1.22
C_200—250	8.5	26.0	74.0	0.0	0.03	0.39
C_350—400	8.4	16.6	83.0	0.5	0.03	0.75

编号	SAs/(ng·g^–1)	FQs/(ng·g^–1)	TCs/(ng·g^–1)	MLs/(ng·g^–1)	平均浓度/(ng·g^–1)	总浓度/(ng·g^–1)
1^[35]	0.50 ~ 276.64		0 ~ 46.10	0.06 ~ 1.51	11.93 ~ 32.86
2^[36]	< 10		1335.24			468.79 ~ 2984.64
3^[37]	53.68 ~ 155.24
4^[38]	3.00 ~ 8.54	28.42 ~ 119.57	ND
5^[39]	2.2 ~ 12.2			0.9 ~ 16.7
6^[40]			80 ~ 6006
7^[40]			84 ~ 1421
8	4.94 ~ 89.09	8.49 ~ 318.10	7.24 ~ 458.77	0.20 ~ 0.57	531.95	38.45 ~ 866.37

土壤层	SAs_ARGs	FQs_ARGs	MLs_ARGs	TCs_ARGs
B1	5.23 × 10³	2.40 × 10⁴	1.44 × 10⁵	1.26 × 10⁵
B2	4.08 × 10³	2.11 × 10⁴	1.05 × 10⁵	0.94 × 10⁵
B3	3.68 × 10³	2.03 × 10⁴	0.92 × 10⁵	0.77 × 10⁵

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