Attenuation of antibiotic resistance genes in livestock manure through vermicomposting via Protaetia brevitarsis and its fate in a soil-vegetable system
Graphical abstract
Introduction
Land application of livestock manure is a common waste process that improves soil fertility and increases food production. However, with the widespread use of antibiotics in animal husbandry, concerns about the antibiotic residues and selective pressure on indigenous antibiotic resistance bacteria are gaining global attention. Accumulating evidence shows that animal manure has been identified as a significant contributor and reservoir of antibiotic resistance genes (ARGs) (Barrios et al., 2021; Leclercq et al., 2016). Previous studies indicated that land application of animal manure without well treatment resulted in a significant rise in the diversity and abundance of ARGs (Zhu et al., 2013; Zhou et al., 2019), posing a potential risk of spreading antibiotic resistance in environment. Understanding the pathway of ARGs dissemination via soil and plant will be necessary for assessing the potential spread of antibiotic resistome in manure.
It has been suggested that ARGs may be transferred from atmosphere aerosol to the plant surface via the external pathway, or through the plant tissues via the internal pathway from manure-amended soils to plants, resulting in a higher degree of ARGs in edible parts (Marti et al., 2013; Chen et al., 2017; Zhu et al., 2017; Zhang et al., 2019). As a result, manure-derived ARGs may enter into the food chain, posing a risk to human health via consuming raw vegetables or direct contact (Rossi et al., 2014; Udikovic-Kolic et al., 2014; Chen et al., 2019). Recent studies have identified a few core microbiota presented across all plant tissues including rhizosphere and phyllosphere (Chen et al., 2021), indicating the capacity of manure-borne ARGs transmission from soil to plant. It is therefore imperative to develop effective strategies to reduce the dissemination risk of ARGs derived from manure for safer utilization of animal manure as fertilizer.
Strategies have been devoted to deal with the manure derived from livestock feedlots like vermicomposting, which is considered as a low-cost and sustainable bio-conversion process for animal manure recycling with potential attenuation capacity in ARGs (Zhang et al., 2012; Bhat et al., 2018; Xia et al., 2019). Vermicomposting via saprophagous fauna, such as earthworms and housefly larvae, resulted in a substantial reduction in the relative abundance of ARGs (Wang et al., 2015a; Wang et al., 2017; Cui et al., 2018; Chao et al., 2019; Liu et al., 2020). The extent to which manure is reduced more efficiently depends on manure type and fauna species (Miranda et al., 2021). Scarab larvae, such as Protaetia brevitarsis, as one of the typical saprophagous faunas, has the ability in digesting organic waste and thus improving soil fertility and sustainability (Ahrens et al., 2014; Wang et al., 2019). P. brevitarsis larva is a common soil insect, feeding on a wide range of organic resources (Li et al., 2019). We therefore hypothesized that the abundance of ARGs in the compost from P. brevitarsis larvae may be reduced, while the fate of frass-derived ARGs in soil-vegetable system remains unknown.
In this study, we investigated the change of ARGs in swine and chicken manure during one-week vermicomposting via scarab larvae as well as the fate of ARGs in the soil-vegetable system through a pot experiment using high-throughput qPCR and Illumina sequencing technique. We aimed to (i) compare the change of ARGs in swine and chicken manures vermicomposting via P. brevitarsis larvae; (ii) assess the impacts of frass application on the spread of ARGs in lettuce. These findings would provide a practical ARGs mitigation approach for livestock manure and highlight their potential risks in soil-vegetable system.
Section snippets
Soil sampling and livestock manure preparation
Soil sampling sites were located at Xingtai county (XT) (27°15′ N, 113°54′ E) and Changfeng county (CF) (28°13′ N, 113°30′ E) in Hunan province, China. These two soils developed from quaternary red earth were collected in April 2018. Five points at top layers (0–20 cm) within each land were taken and mixed into one composite sample as described previously (Zhao et al., 2020). Soil samples were air dried and passed through a 5-mm mesh for the subsequent pot experiment. Swine manure was collected
Changes of ARGs and bacterial community from livestock manure to larval frass
A total of 234 ARGs and 10 MGEs were detected in the livestock manure and larval frass. The detected numbers of ARGs and MGEs in initial swine and chicken manure were both significantly higher than those in the frass (Fig. 1a and b). Venn diagrams showed that 82 and 93 ARG and MGE subtypes were removed from the initial swine and chicken manure samples, respectively. The detected unique ARGs and MGEs numbers in frass samples were significantly lower than those in initial manure samples (Fig. 1c
Attenuation of ARGs in manure through larvae conversion
In this study, frass samples had a significant lower number of ARGs than raw manure samples, indicating that the scarab larva has a great potential in reducing manure-borne ARGs through vermicomposting. Previous studies have found that high efficiency in ARGs attenuation by earthworms and housefly larvae, which further confirmed that vermicomposting has its priority in reducing ARGs compared with traditional composting practice (Wang et al., 2015a). Our previous research has suggested that
Conclusions
In summary, the relative abundance and numbers of ARGs in livestock manure was significantly attenuated through vermicomposting via P. brevitarsis larvae, resulting in a low risk for land application. It has been widely acknowledged that livestock manure vermicomposting via earthworms or housefly larvae may be an ecofriendly technique and is widely applied in industrial sludge treatment and municipal solid waste treatment (Soobhany, 2018; Swati and Hait, 2018). The addition of frass in pot
CRediT authorship contribution statement
Xiang Zhao: Conceptualization, Methodology, Investigation, Formal analysis, Writing – original draft, Visualization. Ju-Pei Shen: Conceptualization, Validation, Formal analysis, Writing – review & editing, Supervision, Project administration. Chang-Long Shu: Conceptualization, Resources. Sheng-Sheng Jin: Methodology, Resources. Hong J. Di: Writing – review & editing. Li-Mei Zhang: Conceptualization, Validation. Ji-Zheng He: Conceptualization, Validation, Writing – review & editing, Supervision,
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
This work was financially supported by National Key Research and Development Program of China (2017YFD0801500 and 2017YFE0109800), and National Natural Science Foundation of China (32070511). Many thanks for Dr. Phillip Chalk from Melbourne University for their helpful comments on this manuscript.
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