猪场废水与周边水环境中细菌和耐药基因的特征

doi:10.3969/j.issn.1004-1524.20231348

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (3): 621-632.DOI: 10.3969/j.issn.1004-1524.20231348

猪场废水与周边水环境中细菌和耐药基因的特征

赵嘉豪¹^,²(), 徐杏², 周卫东²^,^*(), 杨华³, 赵喜红¹, 汪雯³^,^*()

1.武汉工程大学环境生态与生物工程学院,湖北武汉 430205
2.浙江省农业科学院畜牧兽医研究所,浙江杭州 310021
3.浙江省农业科学院农产品质量安全与营养研究所,浙江杭州 310021

收稿日期:2023-11-29 出版日期:2025-03-25 发布日期:2025-04-02
作者简介:赵嘉豪(1999—),男,河南郑州人,硕士,主要从事水环境中微生物检测和杀灭研究。E-mail:febmystery@163.com
通讯作者: * 汪雯,E-mail: ww_hi1018@163.com
周卫东,E-mail: zhouwd@mail.zaas.ac.cn
基金资助:
浙江省重点研发攻关计划(2024C04018);浙江省重点研发攻关计划(2022C02045-3);浙江省农业重大技术协同推广计划(2023ZDXT12-03);研究生教育创新基金项目(CX2022563)

Characteristics of bacteria community and antibiotic resistance genes in piggery wastewater and surrounding water environment

ZHAO Jiahao¹^,²(), XU Xing², ZHOU Weidong²^,^*(), YANG Hua³, ZHAO Xihong¹, WANG Wen³^,^*()

1. School of Environmental Ecology and Bioengineering, Wuhan Institute of Technology, Wuhan 430205, China
2. Institute of Animal Husbandry and Veterinary, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Institute of Agro-Product Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2023-11-29 Online:2025-03-25 Published:2025-04-02

摘要/Abstract

摘要：

生猪养殖废水中含有大量可能的致病菌和抗生素耐药基因(ARGs)。为此,从某生猪养殖场废水生化出水口、猪场附近农田沟渠、猪场内地下水储水罐和池塘中采集样本(分别简记为MW、IW、UW、PW),分析猪场废水和周边水环境中水质、细菌群落和代表性ARGs的特征,并利用16S rRNA测序和实时荧光定量PCR分析了细菌群落和代表性ARGs的丰度。结果表明,MW的化学需氧量、氨氮含量、总磷含量显著(P<0.05)高于其他样本,ARGs总丰度最高;UW的水质最好,菌群的多样性和丰富度最低。从4种水体中均检出不动杆菌属(Acinetobacter)、梭状芽孢杆菌属(Clostridium_sensu_stricto_1)和假单胞菌属(Pseudomonas)等致病菌,且MW和IW中的主要ARGs均为四环素类(tetQ和tetW)和大环内酯类(ermB)耐药基因。典型相关分析的结果表明,猪废水菌群中的菌属为主要的ARGs宿主,其相对丰度与多种检测的ARGs的丰度呈显著正相关。综上所述,MW的高污染指数、多样致病菌和高丰度的ARGs可能对周边水环境构成严重威胁。因此,猪场废水处理需严格执行相关排放标准,且废水最终的深度消毒也必不可少。

关键词: 猪场废水, 细菌群落, 耐药基因, 水质参数, 水环境

Abstract:

The piggery wastewater might be an important reservoir of pathogenic bacteria and antibiotic resistance genes (ARGs) in the environment. In the present study, the characteristics of water quality, bacteria community and ARGs in swine wastewater and surrounding water environment were explored. Samples were gathered from the effluent from biochemical treatment of piggery wastewater, the irrigation ditch near the farm boundary, the storage tank of underground water, and the pond in pig farm, and were marked as MW, IW, UW, PW, respectively, for short. Then, a series of water quality parameters were analyzed, and 16S rRNA sequencing and real-time quantitative PCR was used to research the bacteria community and the abundance of ARGs in each sample, respectively. Results indicated that the chemical oxygen demand, ammonia nitrogen content and total phosphorus content were significantly (P<0.05) higher in MW than the others, as well as its absolute abundance of detected ARGs. The UW had the best water quality and the fewest bacterial diversity and richness among the four samples. The pathogens, i.e. Acinetobacter, Clostridium_sensu_stricto_1 and Pseudomonas, were detected in these four samples, and the major ARGs were tetracycline (tetQ and tetW) and macrolide (ermB) in MW and IW. Moreover, the correlation analysis revealed that the bacteria in the piggery wastewater were the primary hosts of ARGs, of which the relative abundance presented positive correlations with the absolute abundance of the most detected ARGs. Overall, the high water pollution indices, pathogen diversity, and ARGs abundance in MW might threaten the surrounding water environment of pig farm, thus essential emission on standard and further disinfection procedures should be implemented for piggery wastewater treatment.

Key words: piggery wastewater, bacterial community, antibiotic resistance genes, water quality parameter, water environment

中图分类号:

X508

赵嘉豪, 徐杏, 周卫东, 杨华, 赵喜红, 汪雯. 猪场废水与周边水环境中细菌和耐药基因的特征[J]. 浙江农业学报, 2025, 37(3): 621-632.

ZHAO Jiahao, XU Xing, ZHOU Weidong, YANG Hua, ZHAO Xihong, WANG Wen. Characteristics of bacteria community and antibiotic resistance genes in piggery wastewater and surrounding water environment[J]. Acta Agriculturae Zhejiangensis, 2025, 37(3): 621-632.

图/表 9

表1 各水样的水质参数

Table 1 Water quality parameters of water samples

参数Parameter	UW	IW	PW	MW
Tur/NTU	0.1±0.1 c	0.7±0.1 b	0.7±0.1 b	43.5±1.3 a
COD_Cr/(mg·L^-1)	—	49±3 b	53±5 b	430±14 a
COD_Mn/(mg·L¹)	5.37±0.20	—	—	—
NH₃-N含量NH₃-N content/(mg·L^-1)	1.1±0.1 c	6.9±0.2 b	3.9±0.1 b	46.7±0.8 a
总磷含量Total P content/(mg·L^-1)	0.03±0.01 c	0.95±0.03 b	1.06±0.03 b	19.69±0.11 a
$\mathrm{NO}_3^{-}$含量$\mathrm{NO}_3^{-}$ content/(mg·L^-1)	0.09±0.01 d	2.71±0.01 a	1.10±0.07 b	0.54±0.03 c
$\mathrm{SO}_4^{2-}$含量$\mathrm{SO}_4^{2-}$ content/(mg·L^-1)	0.46±0.01 d	8.76±0.25 c	36.28±0.52 b	126.18±7.35 a
K含量K content/(mg·L^-1)	8.9±0.1 d	10.9±0.2 c	11.8±0.1 b	60.5±0.6 a
Na含量Na content/(mg·L^-1)	72±1 c	178±1 b	179±2 b	195±1 a
Ca含量Ca content/(mg·L^-1)	25.5±1.3 d	46.2±1.2 b	31.5±0.8 c	161.6±3.4 a
Mg含量Mg content/(mg·L^-1)	7.7±0.1 d	14.4±0 b	13.7±0.1 c	23.9±0.2 a
Fe含量Fe content/(mg·L^-1)	—	—	—	8.90±0.84
Cu含量Cu content/(mg·L^-1)	—	—	—	0.63±0.02
Zn含量Zn content/(mg·L^-1)	—	—	—	4.25±0.04
Pb含量Pb content/(mg·L^-1)	—	—	—	0.02±0.01

图1 各水样中细菌群落的Chao1指数和香农指数 “*”和“**”分别表示差异显著(P<0.05)和极显著(P<0.01)。

Fig.1 Chao1 index and Shannon index of bacteria in water samples “*” and “*” indicate significant difference at P<0.05 and P<0.01 level, respectively.

表2 各水样中门水平上的细菌相对丰度

Table 2 Relative abundance of bacteria at the phylum level in water samples

菌门Phylum	UW	IW	PW	MW
变形菌门Proteobacteria	90.79	66.02	45.94	12.20
厚壁菌门Firmicutes	2.75	18.36	12.23	72.03
放线菌门Actinobacteria	3.50	4.57	28.42	3.83
拟杆菌门Bacteroidota	1.00	3.44	5.07	6.08
未定义菌门	1.67	1.62	3.86	1.49
unidentified_Bacteria
弯曲菌门Campylobacterota	<0.01	2.89	1.09	0.70
纤细菌门Gracilibacteria	1.25	<0.01	<0.01	—
广古菌门Euryarchaeota	0.01	0.0 1	<0.01	2.19

图2 各水样中细菌属水平上的相对丰度

Fig.2 Relative abundance of bacteria at genus level in water samples

图3 各水样中属水平上细菌群落的线性判别分析(LDA)

Fig.3 Linear discriminant analysis (LDA) of bacteria taxa at genus level in water samples

图4 各水样中耐药基因的绝对丰度

Fig.4 Absolute abundance of antibiotic resistance genes in water samples

图5 基于典型相关分析(CCA)的水样中细菌群落和环境因子的关系 $\mathrm{NO}_3^{-}$, $\mathrm{NO}_3^{-}$含量;Na, Na含量;$\mathrm{SO}_4^{2-}$, $\mathrm{SO}_4^{2-}$含量;Ca, Ca含量;K, K含量;TP,总磷含量;Mg, Mg含量;NH3-N, NH3-N含量;Tur,浊度;CODCr,化学需氧量。

Fig.5 Influence of environmental factors on bacterial communities based on canonical correspondence analysis $\mathrm{SO}_4^{2-}$, $\mathrm{SO}_4^{2-}$ content; Na, Na content; $\mathrm{SO}_4^{2-}$, $\mathrm{SO}_4^{2-}$ content; Ca, Ca content; K, K content; TP, Total P content; Mg, Mg content; NH3-N, NH3-N content; Tur, Turbidity; CODCr, Chemical oxygen demand.

表3 环境因子对细菌群落影响的显著性分析结果

Table 3 Results of significance analysis of environmental factors to bacteria communities

参数 Parameter	CCA1	CCA2	R²	P值 P value
K含量K content	0.475 32	-0.879 82	0.967 16	0.008 0
Na含量Na content	-0.23 02	-0.973 14	0.561 16	0.005 0
Ca含量Ca content	0.511 41	-0.859 33	0.991 39	0.000 5
Mg含量Mg content	0.247 545	-0.968 88	0.939 28	0.000 5
$\mathrm{SO}_4^{2-}$含量	0.596 55	-0.802 57	0.879 73	0.011 0
$\mathrm{SO}_4^{2-}$ content
$\mathrm{NO}_3^{-}$含量	0.662 42	0.749 14	0.515 97	0.025 4
$\mathrm{NO}_3^{-}$ content
总磷含量	0.274 27	-0.961 65	0.934 11	0.000 5
Total P content
COD_Cr	0.429 35	-0.903 14	0.963 41	0.008 0
NH₃-N含量	0.391 81	-0.920 05	0.971 83	0.007 0
Tur	0.446 05	-0.895 01	0.970 48	0.005 5

图6 属水平上细菌与耐药基因的共现模式蓝色圆圈的大小与菌属丰度成正比,红色线条代表正相关,蓝色线条代表负相关。

Fig.6 Co-occurrence patterns of antibiotic resistance genes and bacterial communities at genus level The size of blue node is proportional to the concentrations of the bacterial genera. The links in red and blue represent positive and negative associations, respectively.

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猪场废水与周边水环境中细菌和耐药基因的特征

Characteristics of bacteria community and antibiotic resistance genes in piggery wastewater and surrounding water environment

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