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FATE OF BIOLOGICAL
CONTAMINANTS DURING
RECYCLING OF ORGANIC
WASTES
This page intentionally left blank
FATE OF BIOLOGICAL
CONTAMINANTS DURING
RECYCLING OF ORGANIC
WASTES
Edited by
KUI HUANG
Professor, School of Environmental and Municipal
Engineering, Lanzhou Jiaotong University, Lanzhou, China
SARTAJ AHMAD BHAT
JSPS Postdoctoral Researcher River Basin Research Center
Gifu University Japan
GUANGYU CUI
Post-doctoral Researcher, State Key Laboratory of Pollution
Control and Resource Reuse, Tongji University, Shanghai,
China
Elsevier
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Contents
Contributors xi
1. Occurrence, fate, and behavior of antibiotic resistance genes

in organic wastes during biological treatments and current
elimination strategies 1
Wenjiao Li, Sartaj Ahmad Bhat, Yongfen Wei, Fusheng Li, and
Shuhei Tanaka
1. Introduction 1
2. Occurrence of antibiotics, ARGs, and MGEs in various organic
wastes 3
3. Fate and behavior of ARGs and MGEs during different organic
wastes treatments 8
4. Anaerobic digestion 9
5. Aerobic composting 14
6. Vermicomposting 15
7. Elimination strategies for ARGs and MGEs during biological treatments
of organic wastes 16
8. Conclusion and perspectives 21
References 22
2. Human pathogenic microorganism in sludge 27

Pilar Suarez, Gloria Gómez, and Gladys Vidal
1. Introduction 27
2. Presence of the human pathogens in the primary and secondary
sludge 31
3. Technologies for sludge sanitation and their contribution to the
sustainable development goals 36
4. Final remarks 40
Acknowledgment 41
References 41
v
vi Contents
3. Potential of entomocomposting toward soil pathogen

suppression 47
Dennis Beesigamukama, María Gómez-Brandón, and
Chrysantus Mbi Tanga
1. Introduction 47
2. Persistent pathogens in compost 51
3. Fate of pathogens in insect-composted wastes 52
4. Mechanisms of pathogen suppression by insect frass fertilizer 54
5. Impact of insect frass fertilizer on soil pathogens 55
6. Other benefits of insect frass 56
7. Research prospects 61
8. Conclusion 62
Acknowledgments 63
References 63
4. Valorization of fish wastes for circular bioeconomy:

A concern toward antibiotic resistance genes (ARGs)
contaminants for environmental safety 71
Ram Kumar Ganguly and Susanta Kumar Chakraborty
1. Introduction 71
2. Methods of fish processing 73
3. Utilization of contaminants free fish wastes in different industrial
sectors 76
4. Antibiotic resistance genes (ARGs): A potential contaminant
to aquatic ecosystem 80
5. Global fish production and building up of circular economy 85
6. Conclusion 92
Acknowledgments 92
Conflict of interest 92
References 93
5. Dynamics of biological contaminants along with microbial

community during vermicomposting 101
Ankita Chowdhury, Anamika Roy, Mamun Mandal, Sujit Das,
Ganesh Kumar Agrawal, Randeep Rakwal, and Abhijit Sarkar
1. Introduction 101
2. Different earthworms for vermicomposting 103
Contents vii
3. Composition and diversity of microbes associated with

earthworms 105
4. Microbial decontamination of environment through earthworms 108
5. Role of vermicompost and associated microbe in plant growth and disease
management 113
6. Future prospects of vermicomposting 115
7. Conclusion 116
References 117
6. Treatment approaches for bio-contaminants in organic

wastes 123
Shivika Datta, Simranjeet Singh, Praveen C. Ramamurthy,
Ruby Angurana, Daljeet Singh Dhanjal, Vaidehi Katoch, Ashwani Kumar,
Dhriti Kapoor, Jaswinder Singh, and Joginder Singh
1. Introduction 123
2. Classification of bio-contaminants 126
3. Major treatment approaches for bio-contaminants in organic
wastes 126
4. Different parameters influencing the treatment
approaches 130
5. Challenges and trends in the removal of bio-contaminants 133
6. Conclusions and perspectives 136
References 136
7. Fate of bio-contaminants in organic wastes during

composting and vermicomposting processes 143
Fu-Sheng Sun and Guang-Hui Yu
1. Introduction 143
2. Bio-contaminants in organic wastes during thermophilic
composting 144
3. Bio-contaminants in organic wastes during vermicomposting 146
4. Bio-contaminants in organic wastes during coupling composting
and vermicomposting processes 149
5. Application of synchrotron-radiation-based spectromicroscopies in
identifying the binding sites of heavy metals in organic wastes 150
6. Conclusions 153
Acknowledgments 154
References 154
viii Contents
8. Metagenomic analysis revealing the dual microbial community

features in three common vermicomposts 157
Kui Huang, Wenhui Liu, and Hui Xia
1. Introduction 157
2. Methods 159
3. Results and discussion 162
4. Conclusion 172
Acknowledgments 172
References 172
9. Solid waste management during the COVID-19 pandemic 177

Guangyu Cui and Sartaj Ahmad Bhat
1. Introduction 177
2. Global concerns on COVID-19 waste management 178
3. Characteristics of COVID-19 waste 180
4. Management of COVID-19 waste 180
5. Conclusions 183
References 184
10. Effects of bio-contaminants in organic waste products

on the soil environment 187
Gratien Twagirayezu, Kui Huang, and Hui Xia
1. Introduction 187
2. Categories of bio-contaminants found in soil 188
3. Determination methods of bio-contaminants in soil 198
4. Contamination processes 200
5. Soil bio-contamination in different regions 202
6. Perspectives and mitigation measures 205
7. Conclusion 206
Acknowledgments 206
References 207
11. Fate of bio-contaminants in soil systems and available

remediation methods 213
Sartaj Ahmad Bhat, Guangyu Cui, Wenjiao Li, Fuad Ameen, Naik Yaseera,
Yongfen Wei, and Fusheng Li
1. Introduction 213
Contents ix
2. Occurrence of bio-contaminants from manure, sludge,

and wastewater to soil systems 215
3. Remediation of bio-contamination in soil systems 219
4. Conclusions and future prospectus 222
References 223
12. Antibiotics and antibiotics resistance genes in poultry

and animals’ manure and their effects on environment
and human health 229
Tasneem Sarwar and Sardar Khan
1. Introduction 229
2. Types of animal’s manure 236
3. Application of poultry and animal manures 236
4. Antibiotics 240
5. Antibiotics resistance genes in poultry and animal’s manure 243
6. Source of resistance genes in poultry and animal’s manure 245
7. Conclusion 247
References 247
Further reading 258
13. Vermicomposting as a means of removing antibiotic

resistance genes (ARGs) from soil and water 259
Saikat Ranjan Das, Souvik Dey, Abhijit Pradhan,
Bitish Kumar Nayak, Erukala Venkatramaiah, and Dibyendu Chatterjee
1. Introduction 259
2. Vermicomposting process and raw materials used 261
3. Role of earthworms in mitigating ARGs in soil 263
4. Factors directly or indirectly affecting the fate of ARGs during
vermicomposting 265
5. Potential environmental risks of ARGs during vermicomposting 269
6. Control strategies for ARGs during vermicomposting 269
7. Precaution to be taken during vermiremediation 271
8. Conclusion 273
References 273
14. Insight on the prevalence of pathogens present in the municipal

solid waste of sanitary landfills, dumpsites, and
leachate 279
Rituparna Addy, Ajay Kalamdhad, and Vaibhav Vasant Goud
1. Introduction 279
x Contents
2. Sources, transmission route, and fate of pathogens in wastes and

leachates 282
3. Vectors and hosts of viruses 285
4. Infectivity and pathogenicity of viruses 286
5. Identification and characterization of viruses 288
6. Effect on environment and human health 289
7. Management and treatment of MSW 290
8. Future challenges 292
References 293
15. Bioconversion of hazardous organic wastes using

invertebrates 297
Angie M. Pedraza Torres and Juan C. Sanchez-Hernandez
1. Introduction 297
2. Hazardous solid organic wastes 300
3. Vermicomposting 309
4. Insect-based bioconversion 327
5. Biochar: An ingredient to improve hazardous waste bioconversion 339
Acknowledgments 344
References 345
16. Effects of plants’ density on the reduction of antibiotic

resistance genes in vermi-wetland for recycling excess
sludge 359
Kui Huang, Chunlei Sang, Jing Yang, and Hui Xia
1. Introduction 359
2. Materials and methods 362
3. Results and discussion 367
4. Conclusions 375
Acknowledgments 376
References 376
Index 381
Contributors
Rituparna Addy
Centre for the Environment, Indian Institute of Technology Guwahati, Amingaon, North Guwahati, Assam,
India
Ganesh Kumar Agrawal

Global Research Arch for Developing Education (GRADE) Academy Pvt. Ltd., Birgunj, Nepal
Fuad Ameen
Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia
Ruby Angurana
Department of Zoology, Lovely Professional University, Phagwara, Punjab, India
Dennis Beesigamukama
International Centre of Insect Physiology and Ecology, Nairobi, Kenya; Department of Crop Production and
Management, Busitema University, Soroti, Uganda
Sartaj Ahmad Bhat

River Basin Research Center, Gifu University, Gifu, Japan
Susanta Kumar Chakraborty

Department of Zoology, Vidyasagar University, Midnapore, West Bengal, India
Dibyendu Chatterjee
ICAR National Rice Research Institute, Cuttack, Odisha, India
Ankita Chowdhury
Laboratory of Applied Stress Biology, Department of Botany, University of Gour Banga, Malda, West Bengal,
India
Guangyu Cui
State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai, China
Saikat Ranjan Das

Sujit Das
India
Shivika Datta
Department of Zoology, Doaba College, Jalandhar, Punjab, India
Souvik Dey
Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal, India
Daljeet Singh Dhanjal

Department of Biotechnology, Lovely Professional University, Phagwara, Punjab, India
Ram Kumar Ganguly

Department of Zoology, Vidyasagar University, Midnapore, West Bengal, India
Gloria Gómez
Engineering and Biotechnology Environmental Group (GIBA-UDEC), Environmental Science Faculty &
Center EULA–Chile, Universidad de Concepción, Concepción, Chile
xi
xii Contributors
Marı́a Gómez-Brandón
Grupo de Ecologı́a Animal (GEA), University of Vigo, Vigo, Galicia, Spain
Vaibhav Vasant Goud

Department of Chemical Engineering, Indian Institute of Technology Guwahati, Amingaon, North Guwahati,
Assam, India
Kui Huang
School of Environmental and Municipal Engineering, Lanzhou Jiaotong University; Key Laboratory of Yellow
River Water Environment in Gansu Province, Lanzhou, China
Ajay Kalamdhad
Department of Civil Engineering, Indian Institute of Technology Guwahati, Amingaon, North Guwahati,
Assam, India
Dhriti Kapoor
Department of Zoology, Lovely Professional University, Phagwara, Punjab, India
Vaidehi Katoch
Department of Forensic Science, Lovely Professional University, Phagwara, Punjab, India
Sardar Khan
Department of Environmental Sciences, University of Peshawar, Peshawar; Kohat University of Science and
Technology, Kohat, Khyber Pakhtunkhwa, Pakistan
Ashwani Kumar
Department of Zoology, Doaba College, Jalandhar, Punjab, India
Fusheng Li
Wenjiao Li
River Basin Research Center, Gifu University, Gifu; Graduate School of Global Environmental Studies, Kyoto
University, Yoshida-Honmachi, Sakyo-ku, Kyoto, Japan
Wenhui Liu
Mamun Mandal
India
Bitish Kumar Nayak

Angie M. Pedraza Torres

Laboratory of Ecotoxicology, Institute of Environmental Sciences, University of Castilla-La Mancha, Toledo,
Spain
Abhijit Pradhan
Randeep Rakwal
Global Research Arch for Developing Education (GRADE) Academy Pvt. Ltd., Birgunj, Nepal; Faculty of
Health and Sport Sciences, University of Tsukuba, Ibaraki, Japan
Praveen C. Ramamurthy
Interdisciplinary Centre for Water Research (ICWaR), Indian Institute of Science, Bangalore, India
Anamika Roy
India
Contributors xiii
Juan C. Sanchez-Hernandez
Laboratory of Ecotoxicology, Institute of Environmental Sciences, University of Castilla-La Mancha, Toledo,
Spain
Chunlei Sang
School of Environmental and Municipal Engineering, Lanzhou Jiaotong University, Lanzhou, China
Abhijit Sarkar
India
Tasneem Sarwar
Department of Environmental Sciences, University of Peshawar, Peshawar, Pakistan
Jaswinder Singh
Department of Zoology, Khalsa College Amritsar, Amritsar, Punjab, India
Joginder Singh
Department of Biotechnology, Lovely Professional University, Phagwara, Punjab, India
Simranjeet Singh
Interdisciplinary Centre for Water Research (ICWaR), Indian Institute of Science, Bangalore, India
Pilar Suarez
Fu-Sheng Sun
Institute of Surface-Earth System Science, School of Earth System Science, Tianjin University, Tianjin, China
Shuhei Tanaka
Graduate School of Global Environmental Studies, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto,
Japan
Chrysantus Mbi Tanga

International Centre of Insect Physiology and Ecology, Nairobi, Kenya
Gratien Twagirayezu
Erukala Venkatramaiah
Gladys Vidal
Yongfen Wei
Hui Xia
Jing Yang
Naik Yaseera
Department of Environmental Sciences, Government Degree College Anantnag, Khanabal, Jammu and
Kashmir, India
Guang-Hui Yu
Institute of Surface-Earth System Science, School of Earth System Science, Tianjin University, Tianjin, China
CHAPTER ONE
Occurrence, fate, and behavior

of antibiotic resistance genes
in organic wastes during
biological treatments and current
elimination strategies
Wenjiao Lia,b, Sartaj Ahmad Bhata, Yongfen Weia, Fusheng Lia,
and Shuhei Tanakab
a
b
Graduate School of Global Environmental Studies, Kyoto University, Yoshida-Honmachi, Sakyo-ku,
Kyoto, Japan
1. Introduction
Antibiotics are widely used not only in humans but also in animals to
prevent or treat infections caused by bacteria. However, their overuse and
misuse have caused the emergence and prosperity of antibiotic resistance
bacteria (ARB) and antibiotic resistance genes (ARGs), which led to envi-
ronmental and public health issues and are of great concern worldwide.
According to the World Health Organization (WHO, 2016), the ARGs
problem already claims the lives of 70,000 people worldwide every year.
It is estimated that as many as 10 million people could be dying of
ARGs-related causes every year globally, and about 40% of those deaths will
occur in Asia. The rising levels of ARGs can also hinder progress toward
many of the Sustainable Development Goals, especially those regarding
health, food security, and the environment (FAO et al., 2021). Three pos-
sible reasons for ARGs proliferation in the environment were described by
Lee et al. (2017a) as follows: (1) not fully metabolized antibiotics are signif-
icantly excreted, which account for 30%–90% of the total amount; (2) selec-
tion pressure exerted by antibiotics can facilitate the proliferation of ARB;
and (3) horizontal gene transfer (HGT) between bacteria by mobile genetic
elements (MGEs) can promote the spread of ARGs. Among different
MEGs, integrons were widely investigated and found in approximately
9% of sequenced bacterial genomes (Wang et al., 2020a). Accordingly,
Fate of Biological Contaminants During Recycling 1

of Organic Wastes Copyright © 2023 Elsevier Inc.
https://doi.org/10.1016/B978-0-323-95998-8.00011-X All rights reserved.
2 Wenjiao Li et al.
clarification of the occurrence and behavior of antibiotics, ARGs and MGEs

in various environmental samples, is considered as one of the key points for
mitigating the ARGs proliferation in the environment.
Organic wastes such as activated sludge (Huang et al., 2020b; Wei et al.,
2020), animal manures ARGs (Chen et al., 2021; Tian et al., 2021), and
food wastes (Li et al., 2021; Zhao et al., 2022) have been recognized as
significant reservoirs of ARGs. Treatments for converting organic wastes
to fertilizers or soil modifiers are biological processes involving aerobic
and/or anaerobic bacteria (e.g., anaerobic digestion, aerobic composting,
and vermicomposting) that possess a possibility of being a new spot for
ARGs proliferation. Moreover, the not fully metabolized and improperly
handled antibiotics with related ARB can be also transferred into the wastes
treatment systems (Bhat et al., 2022), leading to higher selection pressure for
ARGs spreading. Furthermore, the antibiotics, ARB, ARGs, and MGEs
persisting in the treated organic wastes can enter the soil or surrounding
water environment after being applied as fertilizers or soil modifiers, thus
affecting the total environment including water resources and the food chain
(Li et al., 2021). The spreading pathways of ARB with ARGs and MGEs, as
well as the antibiotics in the environment, are shown in Fig. 1. Therefore,
Fig. 1 Speading pathways of antibiotics, antibiotic resistance bacteria (ARB) harboring

antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in the
emviroment.
Occurrence, fate, and behavior of antibiotic resistance genes 3
not only the fate and behavior of ARGs in various organic wastes during
treatments but also the occurrence and proliferation potential of ARGs
in the final products after treatments are highly required to be clarified.
However, the comprehensive information on the occurrence, fate, and
behavior of ARGs and MGEs during different biological treatments for
organic wastes as well as the current elimination strategies is not systemi-
cally summarized so far. In this chapter, research progress on ARGs and
HGT-related MGEs in typical organic wastes and their fate and behavior
during different biological treatments were summarized. Additionally, the
current elimination strategies for ARGs and MGEs as well as the possible
mechanisms behind them were also discussed. The critical insights pointed
out here may provide new avenues in the aspect of the elimination of
ARGs and MGEs during biological treatments for organic wastes in further
studies.
2. Occurrence of antibiotics, ARGs, and MGEs in various

organic wastes
Table 1 summarized the recent studies in detecting antibiotics,
ARGs, and MGEs from typical organic wastes. Tetracyclines, β-lactams,
macrolides, quinolones, and sulfonamides with their related ARGs were
detected worldwide in different types of organic wastes, which lined with
the commonly used antibiotics described by Pazda et al. (2019). The pres-
ence of antibiotics is closely associated with ARB and ARGs since the main
mechanisms for bacteria to be resistant are linked with the antibiotics:
(1) efflux pump of antibiotics; (2) modification of antibiotic targets;
(3) inactivation of antibiotics; and (4) enhancement of membrane imper-
meability, as could be seen in Fig. 2. Concentrations of antibiotics varied in
different organic wastes and regions as a result of the variation of applied
types and doses. Antibiotics recorded in sludge from China by Huang et al.
(2020a) were extremely higher than other studies on sludge about 2–4
orders of magnitude. Compared with the activated sludge and animal
manure, food waste such as fruits and vegetables contain fewer antibiotics,
ARGs, and MGEs (Chen et al., 2021; Li et al., 2021). Karkman et al.
(2018) indicated that some bacteria could acquire resistance even under
the very low selection pressure of antibiotics. Cui et al. (2019) reported
a significant difference of tet genes with other targeted genes in domestic
activated sludge and explained it as the reason for selective pressure from
Table 1 Occurrence of antibiotics, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) in various organic wastes worldwide.
Antibiotics ARGs MGEs
Relative Absolute Relative
Absolute abundance abundance abundance
Concentration abundance (copies/16S (copies/g- (copies/16S rDNA
Organic wastes Country Type (μg/kg-dry) Type (copies/g-dry) rDNA copies) Type dry) copies) References
Mixture of China Tetracycline; / tetM; sul1, sul2; / 7.9 104– intl1, / 2.8 102, Wang
food waste sulfamethoxazole; ermB; blaCTX-M 2.5 101 intl2 0.8 104 et al.
and waste erythromycin (2021a)
activated
sludge
Kitchen wastes China Tetracyclines; 13.7–57.2; tetA, tetB, tetC, 1.6 1013– / intl1, 4.2 107– / Zhao et al.
sulfonamides; 0.4–20.9; 7.3– tetG, tetM, tetO, 4.5 1014; intl2, 8.9 1010 (2022)
quinolones; 157.4; 0.3–8.0 tetQ, tetW, tetX; 8.8 108– ISCR1,
macrolides sul1, sul2, sul3, 1.6 1010; IncQ,
dfrA7; qnrB, qnrS, 9.5 107– trbC,
acc(60 )-Ib-cr; 6.4 109; tnpA,
ermB, ermF, 7.0 108– Tn916/
ermQ, ermX, 3.0 1010 1545
mefA
Food waste; China Sulfonamide; 6.4–12.0 in sul1, sul2, sul3 and 5.8 105– / / / / Chen
Sewage sludge fluoroquinolone; food waste, and sulA; qnrB, qnrD 1.4 109 in et al.
macrolide 4.5–19.0 in and qnrS; ermB, food waste; (2021)
sewage sludge ermC and ermF 3.5 103–
3.2 109 in
sewage sludge
Fruit and Japan / / tetG, tetM; sul1 1.0 104– / intl1 3.6 104– N/R Li et al.
vegetable 3.8 106 in 8.1 106 in (2021)
waste; Excess fruit and fruit and
activated vegetable waste; vegetable
sludge 2.1 106– waste;
1.4 108 in 3.8 107 in
excess activated excess
sludge activated
sludge
Municipal China Aminoglycoside; / aadA, strB; ermB, aadA: 2.2 108, / intl1, 4 MGEs: / Tang et al.
solid waste macrolide; ermF; sul2; tetM, strB: 1.3 108; intl2, 5.0 106; (2020)
sulfonamide; tetQ ermB: IS26 intl 1:
tetracycline 6.5 106, ermF: and trbC 1.8 106
2.0 108; sul2:
2.2 108; tetM:
3.4 106, tetQ:
1.9 105
Refuse of China Fluoroquinolone; About 1–1400; qnrD, qnrS, mexF; / 8.2 106– intl1, / About Yao et al.
landfill macrolide; 0.5–130; 1–16 ermA, ermB, 1.3 101; intl2, 103–101 (2020)
sulfonamide mefA; sul1, sul2 5.0 106– tnpA01,
2.2 101; tnpA02,
2.2 103– tnpA03
2.88 101
Dewatered China Tetracycline; 1.9 105; ermF; sul1, sul2; About 8 102– / intl1 About / Huang
sludge norfloxacin; 2.6 106; tetM, tetX 1.3 107 (ng/g) 1.1 104 et al.
enrofloxacin; 2.0 105; (ng/g) (2020a)
sulfanilamide 7.8 105;
Continued
Table 1 Occurrence of antibiotics, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) in various organic wastes worldwide—cont’d
Antibiotics ARGs MGEs
Relative Absolute Relative
Absolute abundance abundance abundance
Concentration abundance (copies/16S (copies/g- (copies/16S rDNA
Organic wastes Country Type (μg/kg-dry) Type (copies/g-dry) rDNA copies) Type dry) copies) References
Excess Japan / / qnrA, qnrS; sul1; About About intl1 About About Cui et al.
activated tetG, tetM, tetX 4.5 105– 1.0 107– 1.4 1010 3.3 103 (2019)
sludge 1.6 1010 3.8 103
Chicken China / / / / / intl1, / About Zhu et al.
manure; intl2 1.5 1015, (2019)
Bovine manure 1.0 1015 in
chicken manure;
About
3.1 1013,
1.2 1012 in
bovine manure
(copies/g-16S
rDNA)
Swine slurry; Spain Lincosamide; N.D.—6600 in sul1; tetW; qurS; / / intl1 / / Gros et al.
Slaughterhouse tetracycline; swine slurry ermB; blaKPC (2019)
sludge fluoroquinolone; solid fraction;
pleuromutilin; N.D.—5100 in
mebendazole; slaughterhouse
macrolide sludge
Manure; Republic Tetracycline; / tetA, tetB, tetE, / 1.4 103– intl1 / / Lee et al.
Sewage sludge of Korea sulfonamide; tetG, tetH, tetM, 1.8 102 (2017b)
quinolone; tetQ, tetX; sul1, in manure;
macrolide; β- sul2; qnrD; ermB, 2.2 105–
lactam ermC; blaTEM 9.3 102
in sewage
sludge
Sludge from United / / sul1, sul2; ermB, About 108–1010 About intl1 About 109 About 102 Ma et al.
wastewater states ermF; tetC, tetG, 102–104 (2011)
treatment plant tetO, tetW, tetX
(primary:
secondary
¼1: 1)
N.D.: Not detected. /: No data.
8 Wenjiao Li et al.
Fig. 2 Main mechanisms for bacteria to be antibiotic-resistant.
the usage of large quantities of tetracycline in clinics, livestock, and poultry

breeding. The original antibiotics and ARB in organic wastes may also
impact the fate and behavior of ARGs during the biological treatment pro-
cess, thus influencing the safety of final products if applied into the soil
environment (Li et al., 2021). Except for antibiotics, a recent review paper
summarized that the occurrence of ARGs in organic waste is also related to
other different factors such as microbial profiles, heavy metals, and chem-
ical properties (Cui et al., 2020). These previous studies evidenced again
that the proliferation of ARGs occurred everywhere with a complex
mechanism, actions are immediately required. Another point worth to
be noted is that studies focusing on both antibiotics and ARGs and MGEs
simultaneously are very limited, remaining unknowns of the interaction of
the occurrence of antibiotics with their related genes.
3. Fate and behavior of ARGs and MGEs during different

organic wastes treatments
Biological treatments, including traditional anaerobic digestion and
aerobic composting, and advanced composting with earthworms (ver-
micomposting), are broadly used for treating organic wastes before disposal
or application into the environment. Our previous publication has summa-
rized the advantages, efficiencies, and final products of these treating tech-
nologies (Li et al., 2020b) and indicated that vermicomposting is one of the
most sustainable methods to treat organic wastes such as fruit and vegetable
wastes and excess activated sludge. However, the evaluation was judged only
based on the utilization value of final products, remaining a lack of safety
issue caused by newly emerging contaminants. Table 2 exhibited the results

and main findings from recent studies on the fate and behavior of antibiotics
(a few studies), ARGs, and MGEs in typical organic wastes during anaerobic
digestion, composting, and vermicomposting. By reviewing previous stud-
ies, new insights on ARGs proliferation during biological treatments involv-
ing bacteria are expected to be gained.
4. Anaerobic digestion
Anaerobic digestion is conducted by involving various bacteria, such
as hydrolytic bacteria, hydrogen- and acid-producing bacteria, and
methane-producing bacteria during different treating phases (Cui et al.,
2020), representing very complex and various changes of microbial commu-
nities. Inoculum of seeding sludge from wastewater treatment plants or pre-
vious anaerobic digesters can also bring about rich bacteria that could be the
host bacteria of ARGs and MGEs, leading to ARGs transfer during digestion
(Wu et al., 2016; Bai et al., 2019; Zhang et al., 2019a). Moreover, anaerobic
digestion consists of mesophilic and thermophilic conditions, which could
also influence the spreading of ARGs. Wu et al. (2016) investigated the fate
and behavior of ARGs and MGEs in municipal wastewater sludge and found
that most ARGs reduced during the acidogenic period and then increased
during the methanogenic period, while the MGEs (intl1) slightly reduced
during both two periods. It seems like acid-forming conditions can play a
role in the elimination of ARGs. They also clarified that the thermophilic
condition at 55°C could mitigate the presence of ARGs compared with
the mesophilic condition at 35°C (Wu et al., 2016). Similarly, a previous
study on anaerobic digestion of dairy manure also revealed that ARGs were
significantly enriched during the mesophilic period at 37°C and then reduced
during the thermophilic period at 55°C, indicating a potential risk of ARGs
spreading in the environment if applied the treated solid fraction. For the
effects of antibiotics on ARGs and MGEs, Bai et al. (2019) conducted
mesophilic anaerobic digestion (35 1°C) to treat dewatered sludge with
and without the addition of six types of antibiotics (2 mg/L) and found that
the enrichment of ARGs in the treatment added with antibiotics was about
twice higher than that in the treatment without addition of antibiotics. These
studies all suggested that higher temperature can effectively inhibit the prolif-
eration of ARGs during anaerobic digestion, while rich ARGs still remain in
the final products or will rebound again if the temperature decreased.
10 Wenjiao Li et al.
Table 2 Fate and behavior of antibiotics, antibiotic resistance genes (ARGs), and mobile
genetic elements (MGEs) in organic waste during different treatment technologies.
Antibiotics
Treatments Organic wastes Conditions Additives Type Before After Type
Anaerobic digestion Municipal (1) Thermophilic: 55°C; Seeding sludge from an / / / 13 ARGs
wastewater sludge (2) Mesophilic: 35°C anaerobic bioreactor
Dewatered sludge Mesophilic (35 1°C); Inoculum sludge from a / / / tetA,

90 days mature mesophilic tetM,
anaerobic digester; tetW;
(1) without addition of sul1, sul2
antibiotics; (2) with
addition of tetracycline,
oxytetracycline,
chlortetracycline,
sulfathiazole,
sulfamethizole and
sulfamethoxazole with a
high concentration of
2 mg/L
Swine manure 37 0.1°C; 30 days Inoculum sludge; / / / 12 ARGs

(1) without magnetite;
(2) with magnetite
Dairy manure Mesophilic (37°C) and / / / / tetA, tetC,

thermophilic (55°C); tetG,
(1) liquid fraction; (2) solid tetO,
fraction with different total tetQ,
solid contents tetT, tetW
and tetX
(1) Aerobic Food waste (1) Inoculum sludge 10 ARGs

fermentation and (anaerobic activated
anaerobic sludge)
co-digestion:
50 days; (2) aerobic
fermentation:
60–80°C for 12–14 h
Composting Kitchen waste (1) 45 days, 55°C (74°C); / Tetracyclines; (1) 0.4–38.9; (1) 0.2–16.2; 21 ARGs
(2) 50 days; 54°C (70°C); quinolones; (2) 4.7–157.4; (2) 0.0–22.1;
(3) 30 days; 40°C (64°C); sulfonamides; (3) 6.1–57.2; (3) 0.0–74.5;
(4) 15 days; 40°C (63°C) macrolides (4) 0.3– (4) 0.0–
13.7 μg/ 31.6 μg/
kg-dry kg-dry
Mixture of food 50 days / / / / 27 ARGs
waste and tobacco
powder (v:v ¼ 3:1)
ARGs MGEs
Before After Type Before After Main findings References
5.6 105– (1) Most of ARGs intl1 About 109 copies/g (1) and (2): The relatively simple Wu et al.
3.3 109 copies/g reduced during slightly reduced microbial community (2016)
acidogenic period with a in with a range of composition during
range from 0.2 to 1.1 log 0.10–0.50 log acidogenic phase of
units while increased unit thermophilic difestion
during methanogenic due to high temperature
period; (2) ineffective in and acid-forming
reducing ARGs conditions played a role
in mitigating the presence
of ARGs
About 1.0 105 copies/ (1) About 1.8 105; intl1 (1) About (1) About Anaerobic digestion Bai et al.
g-dry (2) about 1.9 102; (2) about 5.0 104; showed some enrichment (2019)
3.0 105 copies/g-dry 2.0 102 copies/ (2) about of ARGs in the control
16S rDNA 2.0 105 (1.85 times) and the
copies/16S antibiotics addition group
rDNA (3.05 times). Sludge
digestion at mesophilic
condition could not
effectively remove most
ARGs. the addition of
antibiotics in sludge
could contribute
positively to the
accumulation of total
ARGs in the process of
anaerobic digestion
About 9.0 102 copies/ (1) About 1.6 101; intl1 About 4.5 103 (1) About ARGs and MGEs were Zhang
16S rDNA (2) about 1.3 101– copies/16S rDNA 5.0 103; enriched. Magnetite did et al.
1.7 101 copies/16S (2) about not influence the total (2019a)
rNDA 2.5 103– ARGs abundance
6.5 103
copies/16S
rNDA
(1) 1.2 1010 (1) 3.5 108– intl1, intl2 (1) About (1) About 8.0 ARGs in liquid fraction Zhang
copies/mL; 7.8 109 copies/mL; and Tn916/ 2.7 10 copies/
8
106–9.0 107 decreased by 0.06–1.80 et al.
(2) 9.2 109 (2) 1.5 1010–5.1 1010 1545 mL; copies/mL; logs while those in solid (2021)
copies/g-dry copies/g-dry (2) 6.9 108 copies/ (2) about fraction increased by
g-dry 2.2 108– 0.08–7.85 logs. Digested
9.0 108 solid had the potential
copies/g-dry risks to promote the
spread of ARGs in the
environment, post-
treated is requied before
the application
(1) 6.9 108– (1) 2.7 109–1.1 1010 intl1 and (1) 5.5 1014– (1) 1.0 1016– Total ARG abundance of Wang
3.0 109 copies/mL; copies/mL; intl2 6.8 1015 copies/ 2.4 1017 food waste was increased et al.
(2) 1.3 1010–2.6 1010 (2) 3.8 109– mL; (2) 3.7 1017– copies/mL; after biological treatment. (2020a)
copies/mL 5.5 10 copies/mL
9
2.1 1018 copies/ (2) 1.3 1016– MGEs facilitated ARGs
mL 1.4 1017 propagation
copies/mL
(1) 2.4 1012–2.5 1016; (1) 1.7 1010–3.3 1012;

intl1, intl2, (1) 1.6 1013; (1) and Temperature, pH, and Zhao
(2) 4.2 1011–7.5 1016; (2) 2.5 1011–4.1 1013;
ISCR1, (2) 2.2 1011; (4) declined; water content et al.
(3) 9.5 1010–4.5 1017; (3) 8.9 1011–3.1 1013;
IncQ, trbC, (3) 4.2 1010; (2) and considerably affected the (2022)
(4) 6.4 1012–1.6 1016 (4) 3.7 1011–2.8 1013
tnpA, and (4) 8.9 1013 (3) increased removal of antibiotics and
copies/kg-dry copies/kg-dry Tn916/ copies/kg-dry MGEs
1545
About 2.0 10 copies/ About 1.4 10 copies/ intl1, intl2;
10 11
About About An increase in total Liao et al.
g-dry, 6.5 102 copies/ g-dry, 1.5 102 copies/ ISCR1, 2.0 109 copies/g- 3.0 109 copies/ ARGs and MGEs (2019)
16S rDNA 16S rDNA IncQ; dry, 8.0 103 g-dry, 1.0 104 abundances was
Tn916/ copies/16S rDNA copies/16S observed. Changes in
1545 rDNA ARGs were linked with
shifts in the composition
of bacterial communities
Continued
Table 2 Fate and behavior of antibiotics, antibiotic resistance genes (ARGs), and mobile
genetic elements (MGEs) in organic waste during different treatment technologies—cont’d
Antibiotics
Treatments Organic wastes Conditions Additives Type Before After Type
Mixture of cow 45 days (45°C at day 1, Sulfamethoxazole:(1) 0; Sulfamethoxazole (1) 0.03; (1) N.D.; sul1, sul2,
manure and corn 55°C at day 5, 50°C at (2) 25; (3) 50; (4) 100 (2) 0.27; (2) N.D.; dfrA7,
straw day 11, 45°C at day 21, (mg/kg) (3) 0.82; (3) N.D.; dfrA1, and
40°C at day 31, 35°C at (4) 2.28 mg/kg (4) N.D. after sulA
day 41, and 25°C at day 45) 25 days
Mixture of swine 43 days (55°C for 7 days) / / / / aadA,

manure and straw strB,
(2:1) ermB,
ermF,
qnrS, sul2,
tetM,
tetQ
Vermicomposting Pelleted sewage 2 months; (1) without Cow dung (1: 1 dry basis) / / / 9 ARGs
sludge earthworms (composting);
(2) with earthworms: 100
earthworms/ 4 kg
Pelletized 100 earthworms/ 4 kg; (1) without and (2) with / / / tetC,
dewatered sludge 60 days addition of tetracycline tetG,
with different tetM,
concentrations (100, 500, tetO,
and 1000 mg/kg) tetQ, tetX
(1) Fruit and 10 earthworms/ 100 g; / / / / tetG,

vegetable waste; 25°C; novel vermireactor teteM;
(2) Excess with two compartments sul1
activated sludge;
(3) Their mixture
(w:w ¼ 3:2)
Pelletized excess 60 days; 25 earthworms/ / / / / qnrA,
activated sludge kg-wet; (1) 15°C; qnrS; sul1;
(2) 20°C; 3) 25°C tetM,
tetG, tetX
(1), (2), (3) and (4): different treatments. /: No data.

ARGs MGEs
Before After Type Before After Main findings References
2.8 10 –11
2.8 10 –2.8 10
7 11
intl1 1.4 10 (copies/g) About 1.2 10
11 10
Thermophilic Fahad
6.6 108 copies/g (copies/g) (copies/g) temperature facilitated et al.
the degradation of (2021)
antibiotics; A noteworthy
effect of composting
properties, especially
temperature on bacterial
community, which then
had a positive effect on
ARGs abundances
2.3 109 copies/g-dry 1.1 1010 copies/g-dry intl1, intl2, 3.3 108 copies/g- 1.5 109 Eight target ARGs Guo et al.
(ermB and tetM IS26, and dry copies/g-dry increased 427% on (2021)
decreased, while other six trbC average, with a trend of
ARGs increased) increase at early stage and
decrease at later stage, and
the main reduction stage
appeared in maturity
stage. Prolonging the
maturity period would
inhibit the growth of
potential host bacteria of
ARGs during
composting, therefore
inhibiting the
transmission of ARGs
/ (1) 9 ARGs increased by intl1 / MGEs in final Vermicomposting abated Huang
1–27 folds; (2) 7 ARGs product of 2) was the potential risk of the et al.
(except for tetX and tetG) 68.1% lower ARGs dissemination (2018)
reduced by 40%–100% than those in 1) since the earthworms
strongly affected the
possible host bacteria
encoding ARGs and
MGEs
About 0.0–18.0 (1) tetC and tetG: (efflux intl1 About 1.6 (1) Decreased by Addition of tetracycline Xia et al.
(copies/16S rDNA) pump genes): a small (copies/16S rDNA) 0.7 folds; significantl increased the (2019)
increase; tetM and tetW (2) increased by abundances of ARGs and
(ribosomal protecting 5.1–5.7 folds MGEs. Low tetracycline
genes): small but concentration
insignificant decrease; (100 mg/kg) could
tetX (enzymatic enhance the earthworms’
inactivator): reduced growth
significantly; (2) ARGs in
all treatments were
enhanced
(1) 1.0 104–3.8 106; (1) significantly increased; intl1 (1) 3.6 104– Significantly Elimination of ARGs and Li et al.
(2) 2.1 106–1.4 108; (2) significantly 8.1 106; decreased in the MGEs were probably due (2021)
(3) between (1) and decreased; (3) elimination (2) 3.8 10 ;
7
most treatments to the gut of earthworms
(2) copies/g-dry effect differed among the (3) between (1) and selectively digest the host
studied types of fruit and (2) copies/g-dry bacteria
vegetable wastes
6.0 105– (1) and (2) slightly intl1 2.3 105– Decreased under Vermicomposting at Cui et al.
1.3 109 copies/g-dry decreased; (3) no changes 2.7 107 copies/ (1), (2) and (3) 25°C did not significantly (2022)
or silghtly increased g-dry attenuate the targeted
ARGs relative to that at
15°C and 20°C
5. Aerobic composting
Traditional composting under aerobic conditions is also recognized as
a sustainable technology for treating organic wastes worldwide. It can be
applied for a full treatment course or combined with other treatment
approaches such as anaerobic digestion. Wang et al. (2020b) investigated
the ARGs and MGEs in the initial and biologically treated food waste after
aerobic and anaerobic co-digestion processes and found that the absolute
abundances of ARGs and MGEs decreased obviously after the aerobic pro-
cess while enriched after the anaerobic process. However, the total relative
abundances of ARGs and MGEs significantly increased in both treatments.
The usage of inoculum sludge containing rich ARGs could be one of the
possible reasons for the higher enriched ARGs after anaerobic co-digestion.
A recent study comprehensively confirmed the fate and behavior of antibi-
otics, ARGs, and MGEs in kitchen wastes during composting under differ-
ent temperatures (Zhao et al., 2022). It is recorded that abundances of some
targeted ARGs increased with different extents between a range of 5.4%–
8534.7% and the reduction of antibiotics differed among different types.
Although the kitchen waste was composted, however, it persisted under-
estimated sources of antibiotics and ARGs. Ezzariai et al. (2018) also
reported that reduction efficiencies of antibiotics were different depending
on their chemical structures, while almost could be removed with a range
from 70% to 99%. In addition, Zhao et al. (2022) indicated that temperature,
pH, and water content also considerably affected the reduction of antibiotics
and MGEs. Likewise, Cui et al. (2020) concluded that chemical factors
including pH, NH+4 , and NO 3 could also influence the ARGs in organic
wastes such as excess activated sludge and animal manure. Another more
detailed report of temperature effects on ARGs is given by Fahad et al.
(2021) via carrying out a temperature-controlled aerobic composting of
cow manure mixed with corn straw. ARGs significantly decreased in the
initial 10 days of the thermophilic phase (45°C–55°C) but increased greatly
after 30 days (35°C–25°C). It is designated that the positive effect on the
abundance of ARGs could be explained as the temperature impacted the
bacterial community. The composition of potential host bacteria for ARGs
changed during composting, indicating a close association with the mainte-
nance of ARGs, Liao et al. (2019) also pointed out based on the results
obtained from food waste composting. In contrast to the studies above,
an earlier study on the changes of ARGs and their potential host bacteria
during composting of swine manure described that the maturity period

(lower temperature) is the main stage of ARGs reduction (Guo et al.,
2021). However, the targeted ARGs increased by 427% on average
throughout the composting process. The results from those studies on the
behavior of ARGs and MGEs in organic wastes are inconsistent showing
either decrease or increase during composting, while rich ARGs in the final
product.
6. Vermicomposting
Compared with composting, vermicomposting has more effective
functions of biodegradation and stabilization of the organic wastes through
the joint action of earthworms and microorganisms (Li et al., 2020a). Due to
the joint of earthworms and the bacteria in their gut, the fate and behavior of
ARGs during vermicomposting are more complex than that in composting
because the latter relies only on mesophilic and/or thermophilic bacteria
(Cui et al., 2020). Besides the evaluation based on the stabilization efficiency
of sewage sludge or nutrition values of final products (Huang et al., 2014,
2017), the elimination potentials for antibiotics and ARGs in the sludge
of traditional composting and vermicomposting were also investigated by
(Huang et al., 2018). Through conducting the sludge stabilization experi-
ment with and without earthworms, significant inhibition effects of earth-
worms on the tetracycline and related ARGs, as well as MGEs, were
observed. This phenomenon was explained as the reason that earthworms
can strongly affect the possible host bacteria encoding ARGs and MGEs,
hence abating the pathogenic bacteria in the final product. An earlier study
on treating excess sludge also reported that vermicomposting greatly atten-
uated the targeted ARGs in sludge by 85.6%–100% bringing about the
changes of microbial profiles (Cui et al., 2018). As was mentioned in the
previous sections, temperature plays a critical role in the mitigation of ARGs
and MGEs during biological treatments either under anaerobic or aerobic
conditions, a lack of information for the effect on earthworms and ARGs
in vermicomposting products. Temperature-controlled (15°C, 20°C, and
25°C) vermicomposting of excess activated sludge was established, and
the fate of ARGs was clarified (Cui et al., 2022). Results showed that
25°C cannot significantly attenuate ARGs in sludge compared with lower
temperatures. In addition, the bacterial diversity in the final product was also
impacted presenting a lower Shannon index, indicating that the growth of
some species of possible host bacteria may rebound the ARGs. However, it
is found that the dynamics of targeted ARGs at 15°C and 20°C (higher
kinetic coefficient) fitted to the first-order kinetic equation, proposing that
the vermicomposting at 20°C had more significant removal of ARGs. To
explore the mechanisms of ARGs elimination by earthworms, studies focus-
ing on the bacteria from earthworms’ gut by investigating the fresh casts
were carried out. Cui et al. (2019) clarified that earthworm casts contained
significantly lower absolute abundances of ARGs compared with the initial
sludge before gut digestion. Additionally, the bacterial number and diversity
were remarkably reduced due to the digestion of the earthworm’s gut.
Another recent study investigated the changes of ARGs and MGEs in the
fresh cast derived from earthworms before and after vermicomposting of five
types of fruit and vegetable wastes, excess activated sludge and their mix-
tures, respectively (Li et al., 2021). The relative abundances of ARGs and
MGEs against 16S rDNA in fresh cast samples were reduced markedly
and much lower than those in all different organic wastes after ver-
micomposting (about 1–3 orders of magnitude). It is indicated that the
gut of earthworms could selectively reduce the host bacteria harboring
ARGs and MGEs through activation of non-antibiotic resistance bacteria
or digestion of the ARB.
7. Elimination strategies for ARGs and MGEs during

biological treatments of organic wastes
Previous sections summarized the fate and behavior of antibiotics,
ARGs, and MGEs in typical organic wastes during different biological treat-
ments, as well as possible factors and reasons for the changes of ARGs and
MGEs. Nevertheless, it is worthy to notice that abundant ARGs and MGEs
are still persisting in the final products no matter which treatment technol-
ogy, indicating a possible risk for the proliferation of ARGs in the environ-
ment if applied as fertilizers or soil modifiers. Several elimination strategies
such as the addition of biochar during biological treatments are attracting
researchers’ attention, as summarized in Table 3. Awasthi et al. (2019) eval-
uated the effects of clay amendment in different ratios on the fate of ARGs
and their association with bacterial diversity during composting of poultry
manure. It is figured out that the addition of 4% clay can efficiently mitigate
the rich ARGs in final products since it can alter the physicochemical param-
eters and bacterial composition during the composting of poultry manure.
These findings were in line with a recent study on anaerobic digestion of pig
Table 3 Current elimination strategies used for antibiotics, ARGs, and MGEs in organic wastes.
Fate and behavior of Main findings and possible
Strategies Organic wastes Treatments antibiotics, ARGs and MGEs mehcanisms References
Clay Poultry manure Aerobic composting for Total relative abundance of Addition of clay reduced the Awasthi
mixed with 50 days; (1) without and with
ARGs were considerably ARGs by altering the host et al.
wheat straw addtion of (2) 2%, (3) 4%, reduced and relatively very bacterial species. The (2019)
(5:1, dry basis) (4) 6%, (5) 8%, (6) 10% claylow ARGs was noticed in reduction of ARGs was
(3) followed by (2), (4), significantly associated with
(5) and (6) with a redection the microbial dynamics and
rate of 48%, 42%, 35%, 28% physicochemical parameters
and 16%, respectively
Nanoscale Food waste Anaerobic digestion under ARGs decreased sharply in nZVI changed microbial Wang
zero- mixed with (1) thermophilic: 55°C and all treatments except for communities with a et al.
valent iron anaerobic (2) mesophilic: 35°C treatment (2) and (3); (1) was decreasing the bacteria (2019)
(nZVI) granular sludge conditions; (3) without and more favorable than (2) for belonging to Firmicutes, thus
(2:1, volatile (4) with addition of nZVI the removal of ARGs; resulting in the reduction of
solid basis) (0.1 g/L, 0.5 g/L, 1 g/L, (4) could effectively reduce ARGs
2 g/L, 5 g/L) the ARGs propagation
Nano- Swine manure Anaerobic digestion for Changes in absolute Nano-magnetite showed Zhang
magnetite mixed with 38 days; (1) without and abundances of ARGs and limited effects on the et al.
inoculum (3:1, (2) with the addition of MGEs varied significantly microbial community which (2019b)
basis on total nano-magnetite (5, 75, 150 among the targeted genes. associated with the limited
solid) and 350 mmol) Relative abundances of influences on ARGs and
ARGs and MGEs reduced in MGEs
all treatments. Nano-
magnetite did not avail the
reduction
Continued
Table 3 Current elimination strategies used for antibiotics, ARGs, and MGEs in organic wastes—cont’d
Lignite Pig manure Anaerobic digestion for No significant changes in Addition of lignite Guo et al.
mixed with 30 days; (1) without and with absolute abundances of significantly influenced the (2020)
inocula from addition of (2) 8%, (3) 16%, ARGs and MGEs were microbial community
an anaerobic (4) 32%, and (5) 64% lignite found during digestion for all
compositions, thus
digester based on total solid treatments. ARGs gradually facilitating the removal of
(3:1, w:w) decreased with the increaseARGs and MGEs. The
of lignite. Relative microbial communities
abundances of ARGs and varied in response to
MGEs decreased in differentenvironmental factors
extents among treatments influenced by various
substrates from organic
wastes
Bentonite Food waste Anaerobic digestion under Most of ARGs exhibited Bentonite and temperature Wang
mixed with (1) thermophilic: 55°C and appreciable increases in were key factors that et al.
anaerobic (2) mesophilic: 35°C biogas slurry after treatments impacted ARGs by (2021b)
granular sludge conditions; (3) without and of (4); Both ARGs and influencing the abundance
(2:1, mass basis) (4) with addition of MGEs were reduced in and structure of microbial
bentonite (0.5 g/L, 1 g/L, biogas residue after communities
2 g/L, 3 g/L,5 g/L) treatments (1) and (2), and
(4) enhanced the reduction
of some ARGs and MGEs in
treatment (2)
Biochar Pig manure Stored in a thermostat room Dissipation of antibiotics Dissipation of antibiotics was Ngigi
for 30 days at 20°C; (sulfamethazine, due to adsorption and cation et al.
(1) without and (2) with ciprofloxacin, bridging/exchange, and slow (2020)
addition of 5% biochar (w:w) oxytetracycline and diffusion. Biochar lowered
florfenicol) in treatment the bioavailability of the
(2) was faster and higher than antibiotics, thus leading to
it of treatment (1); ARGs the mitigation of ARGs
increased in both treatments
(1) and (2), while their
abundances were much
lower in (2)
Biochar Dewatered Vermicomposting for Antibiotics significantly Biochar directly affected the Huang
sludge 60 days; (1) without and with decreased as the addition ofmicrobial community et al.
addition of (2) 1.25% and biochar. Changes of ARGs structures due to its porous (2020a)
(3) 5% corncob biochar, and differed among targeted structure and surface, serving
(4) 1.25% and (5) 5% rice ARGs and the types of as habitat for bacteria. It also
husk biochar biochars. MGEs decreased in indirectly affected the
all treatments except for (2)
microbes by impacting
the bacteria inside
earthworm’s gut
Biochar Chicken Aerobic composting for Relative abundances of Reduction of ARGs was Zhou
and zeolite manure and 42 days; (1) without and with ARGs reduced by 92.6%, closely related to the shift of et al.
mushroom addition of (2) 5% zeolite, 95.9%, 98.7% and 98.2% in MGEs. MEGs was greatly (2021)
residues (3) 5% biochar, and (4) 5% (1), (2), (3) and (4), influenced by bacterial
(1:1, w:w) zeolite +5% biochar respectively. Relative community and compost
abundances of MGEs in (2), properties
(3) and (4) were much lower
than those in (1) after
composting
Continued
Table 3 Current elimination strategies used for antibiotics, ARGs, and MGEs in organic wastes—cont’d
Wheat Swine manure Anaerobic digestion for Absolute abundances of Changes of ARGs were Song et al.
straw 55 days; Mixing ratio of ARGs and MGEs decreased associated with the variations (2017)
swine manure to wheat straw by 53.9%, 0.5% and 82.2% in in bacterial compositions and
(1) 3:5; (2) 5:5; and (3) 7:3 (1), (2) and (3), respectively. environmental factors
Relative abundances of
ARGs and MGEs decreased
by 1.03, 1.37, 4.23 logs in
(1), (2) and (3), respectively
Fruits and Excess activated Vermicomposting at 25°C; ARGs and MGEs in (1) and Treating excess activated Li et al.
vegetables sludge (1) without and (2) with (2) reduced significantly with sludge with fruits and (2021)
addition of 5 types of fruits different extents among the vegetables together can
and vegetables (2:3, wet types of fruits and vegetables maintain a better
basis) environment (e.g. carbon to
nitrogen ratio, bacteria
numbers and activities) for
earthworms. Digestion by
earthworm’ gut selectively
inactivated the host bacteria
of ARGs and MGEs
(1), (2), (3), (4), (5), and (6): different treatments.
manure with the addition of another additive namely lignite (Guo et al.,
2020). Guo et al. (2020) also indicated that the addition of lignite facilitated
the reduction of ARGs due to its effects on the shifts of the microbial com-
munity responding to the physicochemical factors. Besides, some advanced
nanomaterials such as nanoscale zero-valent iron (nZVI) and nano-
magnetite were also be explored for the elimination of ARGs during bio-
logical organic wastes treatments (Wang et al., 2019; Zhang et al.,
2019b). No matter the reduction of ARGs and MGEs was enhanced by
the additives or not, a strong interaction among the additives, environmental
factors, and microbial community was found. Biochar is highly porous and
could effectively adsorb the substrates; therefore, it is widely used for con-
taminants controlling in many aspects such as antibiotics and heavy metals.
Recently, the feasibilities of biochar for eliminating the ARGs and MEGs
during the storage of pig manure (Ngigi et al., 2020), vermicomposting
of dewatered sludge (Huang et al., 2020a), and aerobic composting of
chicken manure (Zhou et al., 2021) were also investigated. An agreement
that can be drawn is that biochar can influence the physicochemical factors
of organic wastes, hence further impacting the bacterial composition, which
directly responds to the ARGs and MEGs. Moreover, some researchers have
tried to mix different types of organic wastes to eliminate the ARGs and
MGEs by adjusting the properties of wastes (Song et al., 2017; Li et al.,
2021). The strategies mentioned above can provide some insights for elim-
inating ARGs and MGEs during biological treatments of organic wastes.
However, considerable ARGs and MGEs still can be found in the final prod-
ucts after treatments, and deeper studies are highly required.
8. Conclusion and perspectives

The occurrence, fate, and behavior of antibiotics, ARGs, and MGEs
in typical organic wastes including food waste, sewage sludge, and animal
manure during different biological treatments were summarized. Temper-
ature and the presence of oxygen are two important factors that significantly
affect the shifts of the possible host bacteria harboring ARGs. The joint of
earthworms with their gut bacteria seems could more effectively reduce the
ARGs and MGEs. In addition, the current strategies for the elimination of
ARGs and MGEs in organic wastes such as the addition of biochar were also
discussed and confirmed. The persisting of ARB, ARGs, and MGEs in the
final products after different treatments can bring about environmental and
food chain issues, suggesting that the mobility of ARGs from the final
product to the soil environment or the vegetation needs to be clarified. Fur-

thermore, most of these studies were conducted at the laboratory scale using
a bioreactor, investigations in practical application areas should be carried
out for better understanding and controlling of ARGs spreading.
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CHAPTER TWO
Human pathogenic
microorganism in sludge
Pilar Suarez, Gloria Gómez, and Gladys Vidal
1. Introduction
Sewage is a mixture of black water and gray water. Sewage is made up
of human stool and urine. On the other hand, gray water is mainly com-
posed of detergents from household washing (Bauman-Kaszubska and
Sikorski, 2018). Due to the amount and variety of pollutants present, both
organic matter, nutrients, micro-pollutants, and microbiological compo-
nents, sewage must be treated (Garcia-Aljaro et al., 2019; Praus, 2019).
Fig. 1 shows different stages of treatment that the wastewater must
undergo for purification (WHO, 2016). The first phase is a physicochemical
treatment that seeks to remove suspended solids, fats and oils, generating the
primary sludge as a by-product. Due to this, the primary sludge contains 2%
to 9% solids, while 90% is water (Gherghel et al., 2019).
Subsequently, the wastewater undergoes a biological treatment to elim-
inate organic matter through aerobic or anaerobic biological conditions. In
this stage of the treatment, secondary effluents are generated that still contain
pathogenic microorganisms, which must be cleaned up through different
disinfection technologies. In addition, the aerobic biological treatment
transforms 60% of the organic matter into bacterial biomass, generating
the secondary sludge. Whereas, in anaerobic treatments, only 10% of
the organic matter is transformed into biomass. The secondary sludge is
characterized by having a concentration between 0.8% and 3.3% of total
solids and 90% of water (Gherghel et al., 2019).
Sludge management is a problem for all sewage treatment systems. An
estimated 5.3 million metric tons per year of sludge are produced in the
United States (Dumontet et al., 2001). Sludge production in developing
countries is estimated to vary between 346 and 504 m3/per capita day
(Mesdaghinia et al., 2015). The greater the number of inhabitants, the more
Fate of Biological Contaminants During Recycling 27

of Organic Wastes Copyright © 2023 Elsevier Inc.
https://doi.org/10.1016/B978-0-323-95998-8.00012-1 All rights reserved.
28 Pilar Suarez et al.
Fig. 1 Origin of primary and secondary sludge in a sewage treatment sequence.
by-products are generated. The sludge generated during the wastewater

treatment process corresponds to 2% of the total volume of processed
water (Hawrylik, 2020). This sludge contains more than 50% of the
pollutants present in sewage (Hawrylik, 2019; Venegas et al., 2018,
2021). The most important pollutants in sludge are organic matter, which
corresponds to 60%–70% of the composition of a secondary sludge; It is also
characterized by heavy metals, pathogenic microorganisms such as bacteria,
and emerging compounds (Reyes-Contreras et al., 2018; Venegas et al.,
2021; Wu et al., 2020).
In the sludge, pathogenic bacteria can reach concentrations higher than
their infecting doses in humans. Similarly, other microorganisms such as
viruses and parasites present in sludge can also affect human health in the case
that the sludge would be reused (Li et al., 2022). Furthermore, heavy metals,
emerging pollutants such as antibiotics, antibiotic resistance genes, and
microplastics can be found in the mud (Gao et al., 2020; Venegas et al.,
2021; Zhao et al., 2021).
Sludge must be stabilized through processes that reduce water, stabilize
organic matter, and eliminate pathogens (European Commission, 2001;
Diocaretz and Vidal, 2010; Shoushtarian and Negahban-Azar, 2020). Stabi-
lization processes mainly inactivate pathogens by applying chemical and/or
Human pathogenic microorganism in sludge 29
physical systems. The stabilized sludge is called a biosolid. Once stabilized,

the sludge can be directed to its final disposal: sanitary landfills, incineration,
or direct recycling on the ground according to its characteristics (Zhao et al.,
2019; Fernández et al., 2020).
Sanitary monofills are built facilities, in which the biosolids are
compacted in layers to the minimum volume. Biosolids are not always
completely dehydrated, so there is a risk that, due to percolation or runoff,
the water can spread to the soil. This can have a serious environmental
impact since it could be contaminated soils and groundwater (Zhao et al.,
2019). Despite this, sanitary landfills predominate in developing countries
due to their low cost of operation and maintenance, but in developed coun-
tries, their use is being prohibited (Zhao et al., 2019).
The incinerated sludge is used to produce energy. One of the by-
products is the ash that can be used to apply on the ground (Liu et al.,
2020). Other by-products are CO2, CH4, and N2O, which contribute to
the concentration of greenhouse gases (GHGs) in the atmosphere, which
is highly questioned. Applying an incineration process can generate up to
3124 kg CO2-eq/t dry sludge. However, this practice is unsustainable for
the environment in the context of the current climate change that we have
(Zhao et al., 2020).
Biosolids can be recycled, due to their composition rich in nutrients for
plant growth, so they can be applied in the soil to improve nutritional con-
ditions (Fernández et al., 2020; Li et al., 2022). If biosolids are applied to the
soil, they must be analyzed microbiologically. In this analysis, some regula-
tions require the enumeration of certain pathogenic agents. Thus, in China,
the mortality of 95% of Ascaris sp. Eggs are required, and <0.01 Escherichia
coli cells per liter. In the European Union, the fecal coliform count is
required to be less than 99.99% (1 103 NMP/g dry solids—DS) and Sal-
monella spp. it should not be detectable at 50 g/DS (European Commission,
2001). However, the regulations do not consider other pathogenic organ-
isms, such as viral agents or parasitic protozoa. The problem is that, when
applying sludge with pathogenic microorganisms, these can contaminate
crops, surface and underground water, and even be transmitted via aerosols
to the population (Li et al., 2022).
In this context, a bibliometric analysis of the articles related to this chap-
ter was carried out, using the SCOPUS database. The bibliographic search
was carried out in the article title, abstract, and keywords fields, entering as
search the terms pathogen (or pathogenic), sewage, and sludge. The search
was carried out on November 23, 2021, and included the date range from
30 Pilar Suarez et al.
2000 to 2022. The SCOPUS database identified around 1263 documents.

The exported information derived from the platform included document
title, year, abstract, author keywords, and index keywords. The analysis
of the exported data was performed using “VOSviewer version 1.6.15”,
2020. A co-occurrence analysis was performed, of the 9809 keywords,
243 reached the minimum number of occurrences (CVTS, 2019). Finally,
the 100 keywords that have the most co-occurrence links with other key-
words were selected.
Fig. 2 shows the projection of the research that has been carried out to
increase the knowledge of the characteristics of the sludge generated by sew-
age treatment plants and the study of pathogenic microorganisms present in
them. As described in Fig. 2, four clusters were identified in terms of co-
occurrence. The first cluster (32 items) is based on the sewage sludge con-
cept, this is related to concepts of land uses and associated risks such as agri-
culture, fertilizer, Salmonella, also emphasizing recycling and sludge disposal.
The second cluster (27 items), of wastewater treatment, relates concepts
Fig. 2 Keywords co-occurrence bibliographic network of scientific reports devotes to

“pathogen (or pathogenic),” “sewage,” and “sludge.” The network was constructed
using VOSviewer program, between the years 2000 and 2022. A total of 100 words were
indicated.
Another random document with
no related content on Scribd:
Much incensed, George rode forward with a crimson face, and
saluting, said: “Colonel, that lady is my wife.”
He did not notice the rapid movement of curiosity and excitement
which his announcement made among such of the other officers as
stood near. They had all looked at the girl’s pretty face with surprise
and admiration, but the information that she was the wife of a
comrade produced upon them much the same effect that the
discovery of a fox in an animal they had taken for a rabbit would
have on a group of hunting-men.
But George’s attention was wholly absorbed by the strange
demeanour of Lord Florencecourt, who seemed to have forgotten all
about his marriage, for he started and stared at him with a fierce
amazement utterly bewildering to the younger man.
“Your wife!” he echoed in a low voice. And he faced Lauriston with
a searching look, which the young lieutenant met steadily.
They stood like this for some moments, each finding it hard to
control his seething anger. The Colonel, as became the elder man
and commanding officer, recovered himself first, and told George
curtly that, as parade was over, he had better rejoin his wife and lead
her off the ground. This suggestion the young man was glad enough
to take, and he saluted and rode off without a word, still in a state of
hot indignation. The sight of Massey and Dicky Wood standing
beside his wife, evidently both doing their best to make themselves
agreeable to her, and succeeding to all appearances very well, did
not tend to soothe him, and on reaching the spot where they stood,
he swung himself off his charger in a most unamiable mood. He had
self-command enough left not to reproach her in the presence of his
comrades, but the tone in which he said, without taking any notice of
them, “Come up to my rooms, Nouna,” and the young wife’s sudden
pallor at his words caused the two young fellows to exchange
significant looks, which both Lauriston and his wife unluckily saw,
expressive of fear that the poor little lady was going to have “a bad
time of it.”
At the door of the officers’ quarters they came upon the Colonel,
who was looking as uncompromisingly fierce as ever. He examined
Nouna from head to foot with a straightforward aggressive scrutiny
which made George’s blood boil, while his wife, for her part, stopped
short to return his stare with equally simple directness.
“George, who is he?” she asked suddenly in a low eager voice,
turning to her husband as he put his arm brusquely within hers to
lead her past.
“Lord Florencecourt, the Colonel,” he whispered back, in an
important tone, hoping that the officer’s position would impress her
sufficiently for her to awake to her want of respect.
But before George could see what effect his words might have, the
Colonel himself, who was looking very haggard and grizzly this
morning, an object grim enough to arrest any woman’s attention,
broke into the whispered conversation with brusque coldness. He
had not lost a word of the rapid question and answer, and a slight
change passed over his ashen gray face as if the blood were flowing
more freely again, as he noted the unconcern with which the lady
heard the announcement of his name.
“Pray introduce me to your wife, Lauriston,” he said in such a hard
voice that the request became an abrupt command, without taking
his eyes from her face for one second. “Perhaps, indeed, we have
met before. Mrs. Lauriston seems to know me. In that case I hope
she will pardon my short memory.”
“No, I haven’t met you,” said Mrs. Lauriston hastily, looking at him
with open aversion and turning to take her husband’s arm as if she
considered the hardly-formed acquaintance already too long.
“Then my memory is better than yours, I am sure,” said he, with a
ghastly attempt to assume his usual society manner. “What was Mrs.
Lauriston’s maiden name?” he asked, turning to the young
lieutenant.
“Miss Weston, Nouna Weston,” answered George, with growing
curiosity and interest.
The Colonel’s face remained impassive as wood.
“Ah! Any relation to Sir Edward Weston, the architect?”
“Are you, Nouna?”
“I never heard of him,” said the girl, while her eyes remained fixed,
with the fascination of repulsion, on the Colonel’s hard, lined face.
“My father was Captain Weston, and he died in India; I don’t know
anything else.”
“You came here this morning to see your husband drill?”
“Yes.”
“Ah!” The Colonel seemed to be revolving something in his mind,
and he looked again at Nouna for a moment doubtfully, as if
uncertain whether to ask her another question. However, he
refrained from doing so, and only said, still coldly but with a
perceptible diminution of harshness: “I must apologise for detaining
you, Mrs. Lauriston, but your husband is such an old friend of mine
that I could not resist the temptation of making your acquaintance on
the first opportunity.”
With a formal salute the Colonel retreated, and George hastened
up stairs to his rooms with his wife to take off his uniform. At any
other time Nouna would have found great delight in immediately
trying on his sash and drawing his sword; but the encounter with the
Colonel, while it had one good result in averting her husband’s
displeasure with her for following him to the barracks, had damped
her spirits in a very marked fashion.
“George, how could you say that the Colonel was nice?” she
asked almost before the gentleman in question was out of earshot. “I
think he is the most horribly cold, hard man I ever met. It is quite
right for him to be a soldier; he looked as if he wished I were the
enemy and he could hack at me.”
“Nonsense, child,” said George. “He thought he had met you
before, that’s all. And you looked at him in the same way. Are you
sure you never saw him until to-day?” he asked curiously; for he had
been struck by the puzzled interchange of scrutinizing looks, and
was still rather anxiously in the dark as to the circumstances of his
wife’s life before he met her.
“Quite sure,” said Nouna slowly, looking straight before her and
trying to pierce the gloom of old memories. “I seem to have seen
somebody a little like him, I don’t know when and I don’t know
where, but I am sure I have never before to-day seen him himself.
Why, George, he is too horrid to forget!”
And with a start and a little shiver of dislike, she dismissed the
subject and bounded across the room to play like a kitten with the
ends of her husband’s sash.
CHAPTER XV.
Even in the intoxication of the first few days of married life, George
Lauriston had not forgotten his resentment against Rahas, in whom
he could not fail to see a subtle enemy to his domestic happiness.
On the morning after his marriage he had called at the house in Mary
Street, and was not at all surprised to be told by the servant that Mr.
Rahas had gone away. He insisted on seeing the elder merchant
Fanah, who, however, only confirmed the woman’s statement by
saying that his nephew had gone to France on business of the firm.
It flashed through George’s mind that this sudden journey abroad
might be with the object of visiting Nouna’s mother, with whom it was
plain the young merchant had some rather mysterious undertaking;
but the next moment he rejected this idea, being more inclined to the
opinion that the Countess, for some unknown reason, was anxious
to have it believed that she was further off than in truth she was.
He next went up to Messrs. Smith and Angelo’s offices, saw the
elder partner and laid before him a vigorous remonstrance with
Madame di Valdestillas for employing a foreign scoundrel (as
George did not scruple to call Rahas) who dared not show his face
to the husband, as her messenger to a young wife. The old lawyer
listened as passively as usual, and recommended the indignant
young officer to write to the Countess on the subject.
“And if you will take a word of advice,” ended the old lawyer, his
eyes travelling slowly round the sepulchral office as he rubbed his
glasses, “write temperately, much more temperately than you have
spoken to me. The Condesa is a very passionate woman, and while
she is all generosity and sweetness to those she honours with her
regard, she is liable to be offended if she is not approached in the
right way.”
“I don’t care whether she is offended or not,” burst out George,
with all the righteous passion of outraged marital dignity, “and her
generosity and sweetness are nothing to me. She seems to have a
very odd idea of what a husband should be—” At this point Mr.
Smith, who was smiling blandly in a corner of the office, drew his
mouth in suddenly, with a sort of gasp of horror, which he smothered
as his partner’s eyes, without any appearance of hurry or any
particular expression in them, rested for a moment on his face.
George meanwhile went on without pause,—“if she thinks he will
stand any interference between his wife and himself. She has done
her best to ruin her daughter by her fantastic bringing up——”
“Oh, hush! hush!” interposed Mr. Angelo, while his sensitive
partner absolutely writhed as if it were he himself who was being
thus scathingly censured.
George continued: “But she is quite mistaken if she thinks she can
treat her in the same way now. Nouna is my wife, and if I catch any
other messenger, black, or white, or grey, humbugging about trying
to see her without my knowledge, I’ll horsewhip him within an inch of
his life, if he were sent by fifty mothers!”
A curious incident occurred at this point. There was an instant’s
perfect silence in the room. George was standing with his back to the
door of Mr. Angelo’s private office. No sound was audible but the
nervous scraping of Mr. Smith’s feet on the carpet, and a subdued
clearing of the throat from Mr. Angelo. The young husband was too
passionately excited to take note of either face, and both partners
kept their eyes carefully lowered as if they heard the outburst under
protest. Yet in the pause, without any conscious reason, George
turned suddenly and saw against the glass upper panel of the door,
the outline of a woman’s bonnet, with a small plume of feathers on
the left side. He turned again immediately and faced the lawyers with
an entire change of manner. Feeling a strong conviction now that he
had a larger and a more important audience than he had imagined,
with a flash of self-command he controlled his anger, and spoke in a
firm, clear, earnest voice, each word ringing out as if he were giving
a solemn command.
“Perhaps,” he said, “I am judging too harshly. I mustn’t blame any
one for loving unwisely; I haven’t yet shown that I can love wisely
myself. But I wish Madame di Valdestillas to know—I beg you to let
her know—that I have taken upon myself, with all solemnity before
God, the duty she was not herself able to fulfil, of cherishing Nouna
and shielding her with the influence of a home. I ask you also to beg
her not to send Nouna any more costly presents, like the diamonds
Rahas brought; they only make the poor child long for a chance to
show them off, and it will be years and years before I can put her in a
position to wear them without being ridiculous. I’m sure if Madame di
Valdestillas were to know that, she loves her child much too well not
to see that I am right.”
George paused, keeping quite still. For a moment there was no
sound at all, and both the lawyers refrained from looking at him. At
last with a gentle cough, to intimate that he had well weighed this
speech and that he expected as much weight to be given to his
answer, Mr. Angelo answered:
“I quite understand the integrity of your motives, Mr. Lauriston, and
I believe I may answer for it that the Condesa di Valdestillas, when I
have laid your arguments before her, will respect them as fully as I
do. But I believe it is not now premature for me to confess that
circumstances may arise which will make it not only possible, but
desirable that Mrs. Lauriston should grow accustomed to the wearing
of jewels suitable to a lady not only of position, but of wealth.”
George looked steadily at him, in some perplexity. “The fact is, Mr.
Lauriston,” and Mr. Angelo’s eyes travelled round the room and then
rested for a moment in dull and fish-like impenetrability on the young
man’s face before they continued their circuit, “that Madame di
Valdestillas’s first husband left property to a considerable amount
which he willed to his wife, but with the condition that if she married
again it should go to their daughter. Now Madame di Valdestillas, as
you are aware, has married again, and the property would thus have
fallen to Mrs. Lauriston without question if it had not been for one
circumstance. Some relatives of the late Captain Weston’s have
propounded a later will, benefiting them to the exclusion of his wife
and daughter. Now we have the strongest reasons for believing the
will thus suddenly sprung upon us to be a forgery; but until the trial,
which is to take place shortly, we cannot be absolutely sure of our
case. In the meantime the Countess thought it better not too soon to
hold out expectations which might never be realized. It is quite on my
own responsibility therefore, that I have made this communication to
you. It will explain what would certainly otherwise seem rather
mysterious conduct with regard to the present of jewels to her
daughter.”
“It seems to me rather mysterious still,” said George shortly and
uneasily. This network of strange occurrences and explanations that
seemed to him quite as strange, perplexed the straightforward young
soldier. “I’m not such a fool as not to value money, but frankly I’d
rather be without a great deal than think my wife had a fortune which
would make her independent of me. In any case I ought to have
been told the whole position of things before I married her.”
He took up his hat and after very few more words left the office, on
all points less satisfied than when he entered it. He hated humbug,
and this foreign Countess’s playing with him, even if it arose from
nothing but a woman’s love of little mysteries, was exceedingly
distasteful to him. He had acted in the promptest and most upright
way towards Nouna, such as might have convinced any reasonable
person of his integrity, yet in no respect had he met with
corresponding frankness on her side. If he had been told before of
the young girl’s possibly brilliant prospects, it would have changed
many things for him; now that the suggestion was suddenly sprung
upon him late in the day, he found that he could not adjust himself to
the notion of Nouna rich, distracted in the first flush of the
honeymoon by the startling news that she was a wealthy woman,
with a host of luxurious pleasures at her command outside such
simpler, more domestic happiness as her husband could give her.
And he resolved that, as she did not know of the sensational
prospect that might open before her, he would himself say nothing
about it, but would wait until the will case was tried, and the matter
finally settled in one way or the other.
George Lauriston, having held himself till now rather markedly
aloof from the influence of feminine fascination, was now expiating
his neglect in daily tightening bondage at the feet of the most
irresistible little tyrant that ever captivated a man’s senses and
wormed her way into his heart, none the less that he saw daily with
increasing clearness how much more he was giving her than she
was giving, or perhaps could give, to him. George was puzzled and
disappointed. Arguing from his personal experience, in which the
ecstatic dreams and timid caresses of the lover had been but a weak
prelude to the ardent and demonstrative tenderness of the young
husband, he had taken it for granted that those pretty, capricious
outbursts of girlish passion, which had charmed him so much by
their piquant unusualness, would develop under the sunshine of
happy matrimony into a rich growth of steady affection, coloured by
the tropical glow which seemed to belong to her individuality, and
cherished and fed by his own devotion. It almost seemed sometimes
as if marriage had had the effect of checking her spontaneous
effusiveness, as if she was rather afraid of the violent
demonstrations which any encouragement would bring down upon
her. Now George, at three-and-twenty, could scarcely be expected to
be much of a philosopher; and finding in his own case that true love
was indeed all-absorbing, he saw no reason for doubting the
common belief that true love must always be so. Only at a much
later stage of experience does one understand that into that vaguely
described state of being “in love” enter many questions of race,
complexion, age, sex, and circumstances, which produce as many
varieties of that condition as there are men and women who pass
through it. So he fell ignorantly into the mistake of thinking that he
had not yet succeeded in wholly winning his wife’s heart, and greatly
tormented both himself and her by laborious and importunate efforts
to obtain what was, as a matter of fact, safely in his possession.
Nouna loved her husband as a bee loves the flowers, or a kitten
the warmth of the sun. He was the prince she had waited for to take
her out of the dull twilight of life with Mrs. Ellis and music-lessons;
and although, in the modest nest which was all he could yet make for
her, there were missing many of the elements upon which she had
counted in her imaginary paradise, yet who knew what glories might
not be in store for her in the rapidly approaching time when George
would be a General and wear a cocked hat? And in the meantime he
was the handsomest man in the world, and kinder and sweeter than
anybody had ever been to her; though when she looked into his eyes
and sighed with voluptuous delight at the lights in them, and at their
colour and brilliancy, in truth she read in them little more than any
dog can read in the eyes of his master, and she alternated her
moods of passionate satisfaction in her new toy with moods in which
she openly wished that she were not yet married to him, so that she
might have all the novelty and excitement of the wedding over again.
And George, who in his efforts to resist the temptation of
becoming a mere slave to this little princess’s caprices, ran a risk of
becoming a later and worse Mr. Barlow, decided that, in the shallow
education she had received, the intellectual and spiritual sides of her
nature had been too much neglected, and set about remedying
these omissions in a furiously energetic manner. He was beset by
many interesting difficulties. To begin with: what pursuit could be
imagined so pleasant and at the same time so improving as reading
for filling up the hours during which he was forced to be absent from
her? But Nouna lightly, firmly, and persistently refused to read one
line even of a judiciously chosen novel, although George had taken
care to tempt her by a set of beautifully bound volumes by a lady
writer who took a decorously vague and colourless view of life
through Anglican-Catholic spectacles. She would look out of window
by the hour, lie on the sofa listening to the songs and tales of
Sundran, whom George hated himself for his weakness in not
dismissing, even catch flies on the window-panes and give them to
the cat, anything rather than open a book. She would, indeed, permit
George to read to her, lying curled up in his arms and hearing for the
most part without comment, unless he chose poetry. In this region
she showed marked preferences and prejudices. Shakspeare, she
averred, made her head ache, with the exception of certain chosen
passages, which she would hear again and again, strung together in
odd fashion. She was never tired of the love-scenes in Romeo and
Juliet, but she would not permit the intervening scenes to be read,
preferring a short summary in George’s own words to fill up the
blanks thus made in the story. Othello she would suffer in the same
way, and King Lear, The Merchant of Venice, and The Taming of the
Shrew; and she was never tired of the description of the death of
Ophelia. The Idylls of the King she preferred for the most part
without the poetry, but she learned by heart The Lady of Shalott, The
Moated Grange, and other pieces which presented ladies in a
picturesque and romantic light. Some portions of Hiawatha had a
strong fascination for her, and as, again and again, George read to
her the account of the death of Minne-haha, he would feel her arms
tighten round his neck, and hear her breath come short with intense
interest and emotion close to his ear.
“Would you feel like that if I died?” she asked abruptly one
evening, laying her hand across the page when he had read to the
end.
“My child, my child, don’t ask me,” whispered George, overcome
and thrown off his balance by a sudden realisation of the strong hold
this little fragile woman-creature had got upon his whole nature, of
the paralysing dead blank her absence would now make in his life. “I
don’t think, my wife, that I should live long after you,” he said in a
grave, deep voice, laying his right hand upon her shoulder, and
tightening the clasp of his left arm round her waist.
“Really? Would you die too, like the people in poetry?” she asked,
delighted, rubbing her round young cheek against his in appreciation
of the appropriateness of his answer. But then, examining him at her
leisure, a doubt crossed her mind, and as she spanned the muscles
of his arm with her little fingers she shook her head. “A man is so
strong, so wise, and has so much more to do in the world than a
woman, that I think he could not die off just when it pleased him,”
she said thoughtfully. “There are always Nounas in the world, I think,
just like flies and flowers, and silly useless things like that. If one
goes, one knows it must go, and one does not miss it. And a man
sees that, and he says, ‘Ah, it is a pity!’ and then he goes on living,
and the grass grows up on the grave, and he forgets. After all it was
only a little flower, only a little fly. And so while we are alive we must
just be sweet, we must just fly about and buzz, for when the little
grave is made we don’t leave any trace. That is what I think,” ended
Nouna with a half-grave, half-playful nod.
But George could not take the speech playfully at all. This light but
resolute refusal to take herself in earnest, which he ascribed to the
paralysing influence of Eastern traditions, was the great barrier to all
higher communion between them than that of caresses. By the
expression of his face Nouna scented the sermon from afar, and as
he opened his mouth to speak, she thrust her hand between his lips
as a gag, and continued, laughing:
“Don’t look like that. I won’t hear what you have to say. You may
be very wise, but I must be listened to sometimes. Now, I won’t let
you speak unless you promise to leave off reading and sing to me.”
George nodded a promise, unable to resist her, and there was an
end to the higher education for that evening.
The second string to the young husband’s educational bow was
art, in whose refining and ennobling influence he believed dutifully,
though without much practical sense of it. He took her to concerts in
which she found no acute pleasure, declaring that sitting still so long
in one narrow chair tired her, and that she would rather hear him play
and sing to her at home. He took her to picture galleries, which
would have been a rather penitential exercise for him by himself, but
from which he thought her more delicately organized feminine
temperament might derive some benefit, “taking it in through the
pores” as it were, as boys absorb a love and longing for the hunting-
field from the sight of their father’s scarlet coat and hunting crop.
Now this experiment had more interesting results. The first place he
took her to was the Royal Academy, where she examined the
pictures in a dazed silence, which George hoped was reflective
admiration; but when they returned home she confessed simply that
she did not want to see any more pictures. In the National Gallery,
on the other hand, where George took her rather apologetically, with
a sort of feeling that this was too “advanced” for her present state of
art knowledge, Nouna, at first sight of the frames inclined to be
restive, began speedily to show an odd and unexpected pleasure,
which deepened before certain Gainsboroughs into childish delight.
“I should like the gentleman who painted that lady to paint me,”
she said, when she had gazed long and lovingly at one graceful
bygone beauty.
George explained the difficulties in the way of her wish, but was
highly pleased with the orthodoxy of her taste.
Later experiences, however, gave a shock to this feeling. The
National Gallery having effected her reconciliation with pictorial art,
Nouna was praiseworthily anxious to learn more of it, and insisted on
visiting every exhibition in London. It then gradually became manifest
that she had a marked preference for the works of Continental
painters, from the lively delineator of Parisian types of character to
the works of the daring artist who presents the figures of sacred
history with strong limelight effects.
“They make me see things, and they make me feel things,” was all
the explanation she could give of the instinctive preferences of her
sensuous and poetical temperament.
Even this was not so distressing as her making exceptions to her
indifference to English art in the persons of two artists whom George
had always been accustomed to consider legitimate butts for satire.
The beautiful, mournful women, with clinging draperies, looking out
of the canvas with sadly questioning eyes, imaginative conceptions
of an artist who has founded a school plentifully lacking in genius,
filled Nouna with grave pleasure, and caused her to turn to George
in eager demand for sympathy.
“Eh? Do you like them?” said he, surprised. “Why he’s the man
who started the æsthetic craze; all the women took to starving
themselves, and to going about like bundles of limp rags, to look like
the gaunt creatures in his pictures.”
“That was silly,” said Nouna promptly. “The women in the pictures
mean something, and they don’t care how they look; a woman who
just dressed herself up like them would mean nothing, and would
care only for how she looked.”
George thought this rather a smart criticism, and forgave her
peculiar taste on the strength of it; still, he believed himself to be
quite on the safe side when he said, taking her arm to point out a
picture on the opposite wall by another artist:
“You won’t want me to admire that smudge, I hope, Nouna?”
She remained silent, considering it, and then said gravely:—
“It isn’t a smudge, it’s a lady.”
“Well, do you like it?”
“I like her better than the babies, and ladies, and cows, and
mountains in the Royal Academy.”
“But it takes half an hour to find out what it is.”
“That’s better than making you wish it wasn’t there.”
After that George gave her up, and began to perceive that it would
need a critic more apt than he was to deal aright with her perverse
but intelligent ignorance.
His third means of developing what was noblest in his wife’s
character was, of course, religion. George was not religious himself,
but it seemed a shocking thing for a woman not to be so, and still
more for her to lie under the suspicion of practising the rites of an
occult pagan faith. So he took her to church, where she shook with
laughter at the curate’s appearance and voice, and yawned, and
played with her husband’s fingers during the sermon.
“Oh, George, how clever it was of you not to laugh at the little man
in white!” she cried, with a burst of laughter, at the church-door, when
she had hurried down the aisle with indecorous haste. “Now I’ve
been once to please you, you won’t make me go again, will you? It
reminds me so dreadfully of the horrid Sundays at school.”
“Well, but don’t you like being in the church where we were
married, darling?” he asked gently.
“Oh, but I can remember I’m married to you without going in
again,” she answered laughing.
And so gradually this desultory musical and religious education
dwindled down to visits to Westminster Abbey and the opera; nor
could George succeed satisfactorily in establishing in her mind a
proper sense of the difference there is between these two kinds of
entertainment.
CHAPTER XVI.
The honeymoon was over, and the London season drawing to a
close before the Colonel, who, to Lauriston’s great regret, seemed,
since that inauspicious introduction to Nouna, to have withdrawn into
a permanent coldness towards him, made an attempt to bridge over
the restraint which had grown up between them. It was one evening
when George, to do honour to a visitor who was a friend of his, had
dined at mess, that the Colonel broke silence towards his old
favourite, and inviting him, at dessert, to a chair just left vacant by his
side, asked if he was still as anxious as ever to get leave for
September.
George was rather surprised.
“I am more anxious than ever for it now, sir,” he said. Then, seeing
that Lord Florencecourt’s brows contracted slightly with a displeased
air, he added apologetically, “You know, sir, I should not have
ventured to ask for more leave this year if you yourself had not been
kind enough to propose it. And now my wife is longing for the
promised change.”
The Colonel instinctively frowned still more at the mention of the
obnoxious wife, and after balancing a fruit-knife on his fingers for a
few seconds, with his eyes fixed as intently upon it as if the feat were
a deeply interesting one, he said very shortly:
“You still intend to go to Norfolk? It’s a damp climate, an unhealthy
climate—not one suitable to a lady born in India, I should think.
Beastly dull, foggy place at all times. Why don’t you go to
Scarborough?”
George could not remind him that it was he himself who had
probably suggested the Millards’ invitation, and certainly done his
utmost to persuade him to accept it. The knowledge that it was
disapproval of his wife which had caused this sudden change in the
Colonel’s views, made him suddenly stiff, constrained, and cold.
“I have promised my wife, Colonel; it is too late to change my
plans and disappoint her.”
The Colonel glanced searchingly up at him from the corners of his
eyes, and said almost deprecatingly:
“I see—of course not. I only meant to suggest that the quiet
country life the Millards lead might bore her, and, the fact is, our
place will not be as lively as usual this year. I shall have some
unexpected expenses to meet. I’ve been warned of them, and they
will not be long in coming.” Here he paused, gazing still on the table,
and seeming not so much to watch as to listen for some sign of
comprehension on George’s part. “And so,” he continued, at last
raising his eyes, and speaking in his shortest, bitterest tone, “I shall
have to retrench, and there will be no merrymaking at Willingham
this year.”
“I am very sorry to hear it,” said George, puzzled by a swift
conjecture that he was expected to make some more significant
remark, and wondering what it ought to have been.
He was wishing he could withdraw from this awkward tête-à-tête,
when, in a low distinct voice, the Colonel struck him into perplexity
by the following question:
“How would you like to exchange into the ——th, and go over to
Ireland?”
“Not at all,” answered George, just as low, and very promptly.
He was extremely indignant at this suggestion that he should go
into exile just for having pleased himself in the matter of his
marriage, which was unmistakably his own affair.
“Would you not if by so doing you could confer a very considerable
favour upon one whom you used to be glad to call an old friend?”
said the Colonel in the same low voice, and with a strange
persistency.
Around them the sounds of laughter and of heated but futile after-
dinner discussions, beginning in wine and dying away in cigar-
smoke, filled the hot air and rendered their conversation more private
and at the same time freer than it could have been if held within
closed doors. George looked at the ashy pale face of the
prematurely aged officer, and it seemed to him that his own frame
shivered as if at touch of some unexpected mystery, some unknown
danger. He answered with much feeling:
“Tell me why it is a favour, Colonel. I would do more than this to
show I am grateful to you. Only let me understand.”
But the very sympathy in his tones seemed to startle the Colonel,
who drew back perceptibly, with a hurried glance straight into
George’s eyes. It seemed to the latter, who was now on the alert for
significant looks and tones, that at the moment when their eyes met
the Colonel took a desperate resolution. At any rate, when he spoke
again it was in his usual manner.
“It’s nothing,” he said, waving the subject away with his hand.
“Nothing but a passing freak which I beg you will not think of again.”
His tone notified that the discussion was closed, and for the whole
of the evening George considered, without finding any satisfactory
clue to an explanation, what Lord Florencecourt’s motive could be for
so strongly objecting to Nouna’s appearance in the neighbourhood of
Willingham. His prejudice against swarthy complexions could
scarcely be sufficiently obstinate for him to hope to clear the county
of them: but what was the origin of the prejudice?
On returning home George tried to probe the misty memories
which the Colonel’s appearance had, on his introduction to Nouna,
stirred in her mind. But he could elicit nothing further. Nouna was
now showing at times little fits of petulance, born of the absence of
violent novelty in her life now that the husband was growing to be
quite an article of every-day consumption, as much a matter of
course as dry toast at breakfast, and she was not going to be
troubled to remember or try to remember faces.
“I dare say I only fancied I ever saw anybody like him,” she said
with a little wearied twist of her head and sticking forward of her
round chin. “I can’t count every hair on the head of every old
gentleman I see.”
And however often he might return to this subject, and in whatever
mood she happened to be, George could learn nothing more definite
than her first vague impression, which grew even fainter as the
meeting faded into the past.
In the meantime Nouna was becoming rather weary of looking into
the pretty shop windows without being able to buy anything, and of
walking among the people in the park without joining in any other of
their amusements. George had had designed for her a tailor-made
walking-dress of white cloth embroidered in gold thread and bright-
coloured silks which, with a small white cloth cap embroidered in the
same way, caused her appearance to make a great sensation
among the conventionally ill-dressed crowds of Englishwomen with
more money, rank, and beauty, than taste. He was himself much
surprised to see how easily she wore a dress of a cut to which she
was unaccustomed, and how well she looked in it. The conventional
shape of the gown only emphasised the difference between the
natural movements of her lithe form and the stiff bobbings and
jerkings and swayings which mark the gait of the ordinary English
girl. The reason was simply that Nouna had by nature that great gift
of beauty of attitude and movement which we call grace; and as
among the handsomest women of England only one in every
hundred is graceful either by nature or art, that quality alone would
have been enough to make the half Indian girl conspicuous.
Therefore there was much discussion among onlookers as to her
nationality. The Indian type is not common enough among us to be
widely recognised, or the delicate little aquiline nose, the long eyes,
and the peculiar tint of her skin, might have betrayed her; as it was,
conjectures wavered between France and Spain as a birthplace for
her; for while she wore her dress like a Parisian, she certainly
walked like a Spaniard. By no means unconscious of the attention
she excited, Nouna would have liked to come in closer contact with
some of the handsome Englishmen who seemed by their respectfully
admiring looks to be so well-disposed towards her. For she was
decidedly of a “coming-on” disposition, and not at all troubled with
raw shyness or an excess of haughty reserve. Neither was she
conscious of anything forward or improper in her sociability.
“All these gentlemen that we pass, and those standing in a crowd
near the railings under those trees—they are gentlemen, the English
gentlemen mamma thinks so much of, aren’t they?”
“Oh yes,” answered George smiling, “they are among what are
called the best men, though it’s rather a rum term to apply to some of
them.”
“I should like to know some of them: you do, I see them nod to
you.”
“Wouldn’t you rather know some of the pretty ladies?”
“No,” answered Nouna promptly. “Their dresses always drag on
the ground one side, and they wear dreadful flat boots like Martha
the servant. I like the gentlemen better.”
“But you mustn’t judge people only by their dress, Nouna,” said the
young husband, feeling rather uncomfortable.
“I can’t judge them by anything else till I know them,” said Nouna
fretfully.
George was silent. He was disappointed to learn that she was so
soon weary of the perpetual tête-à-tête which had lost no charm for
him; but he had the sense to own that his ambitions, even the daily
meeting with his comrades at parade, made his life fuller than his
young wife’s could possibly be. He resolved to call upon the Millards,
who, anxious not to intrude upon the newly-married couple before
they were wanted, had refrained from calling upon the bride until
they should hear again from George. On the morning of the day he
had fixed for his call, a note came for him from Captain Pascoe,
asking him to join a party up the river in a couple of days. Nouna
read the note over his shoulder.
“Shall you go, George?” she asked with interest.
“No, dear, how can I? I’ve got a little wife to look after now.”
“But if you were to write and say no, you couldn’t come because
you’d got a wife, perhaps he’d ask me too!”
George had no doubt of this, believing indeed that this was the
result Captain Pascoe had aimed at.
“Captain Pascoe is not quite the sort of man I should like you to
know, dear,” he said.
“Isn’t he an English gentleman?”
“Oh yes, but English gentlemen aren’t all angels, you know.”
“And aren’t you ever going to let me know anybody who isn’t an
angel?”
“Well, darling, I don’t think anybody else is good enough.”
A pause. George hoped she was satisfied for the present. She
was still behind his chair, so that he could not see her face. At last
she asked, in a low, rather menacing tone:
“Are angels’ wings made of feathers, like birds’?”
“I don’t know, dear, I suppose so,” said George laughing, and
turning to look at her and pat her cheek.
“Then if I met one I should pull them out,” she cried in a flame of
fury, and before her husband could recover from his astonishment,
she had fled out of the room.
He followed her with a troubled countenance, and found her face
downwards on the bed, sobbing her heart out. No remonstrances
were of any use, she only murmured that she would like to be a nun,
it was more interesting than to be a wife shut up and never allowed
to speak to anybody.
“But, Nouna, the Indian ladies are much more shut up than you
are.”
“They have beautiful wide palaces to live in. I shouldn’t care if I
had a palace.”
“Well, you know I can’t give you a palace, but if you will be good
and leave off crying, I will take you on the river myself one day.”
“Will you? When, when?” cried she, starting up excited, all her
griefs forgotten.
“I’ll see if I can take you to-morrow.”
She flung her arms round his neck, not to ask his pardon for her
petulance, but to assure him that he was the best, kindest husband
that ever lived, and that no Indian Maharanee in all her splendour of
marble courts and waving palms was ever so happy as she.
George kept his promise, and on the following day took her down
to Kingston, and rowed her up as far as Shepperton and back. She
was delighted with the river, and, charmed with the idea of being a
person of responsibility, showed great aptitude for a beginner at
steering. Being one of those quick-eyed, neat-handed persons
whose wit is rather nimble than profound, she acquired
accomplishments of this nature with a feminine and graceful ease;
and sitting with the ropes over her shoulders, her dark eyes intent
with care gleaming from beneath her white baby-bonnet, she made a
picture so perfect that, as usual, every man who passed looked at
her with undisguised admiration, and glanced from her to her
companion to find out more about her through him. All this George,
who was not too much lost in his own adoration to note the casual
votive glances offered to his idol, bore with complacency, until, just
as they entered Sunbury Lock, on the return journey, a well-known
voice calling his name from a boat that was already waiting inside
the gates, startled him. He turned and saw a crew of four men, two
of whom were Captain Pascoe and Clarence Massey. The
impetuous little Irishman dragged the two boats alongside each
other, and instantly plunged into conversation with Nouna, who
seemed delighted with the incident. George was not a Bluebeard;
still, remembering all the circumstances of Nouna’s previous
acquaintance with the all round lover, Massey, he by no means
desired the friendship to grow closer between them, and he was not
pleased by the glances of interest which Nouna exchanged with
Captain Pascoe, who had an air of quiet good-breeding particularly
attractive to women. The two boats passed each other again and
again on the way to Kingston, for the stronger crew seemed to be in
no great hurry, and were not perhaps unwilling to be occasionally
passed by a boat steered by such an interesting little coxswain. At
any rate the smaller craft arrived first at Bond’s, and George took his
wife up stairs to the coffee-room for a cup of tea. Then she
discovered that she felt rather “faint,” and had forgotten her smelling-
salts; would George go out and get her some? What could a newly-
fledged husband do but comply, however strong his objection might
be to leaving his wife alone in a public room? There was no one in it,
however, but a cheerful and kind little waitress, who seemed quite
overcome by the young lady’s beauty; so he gave Nouna a hurried
kiss when the girl’s back was turned, and hastened off to fulfil her
behest as fast as possible.
He found a chemist’s very quickly, and returned with the smelling-
salts in a few moments. But Nouna had entirely recovered from her
faintness, and instead of finding her reclining on the horse-hair sofa
with closed eyes and a face of romantic paleness, George
discovered her enthroned in an arm-chair, all vivacity and animation,
holding a small but adoring court composed of the crew that had
dogged their progress on the river. Massey was talking the most; but
Captain Pascoe, by virtue of his superiority in years and position as
well as a certain distinction of appearance and manner, was
undoubtedly the most prominent and the most favoured courtier. For
a moment George stopped in the doorway, as a terrible
remembrance of the tale of the genie who locked the lady up in a
glass case flashed into his mind. He dismissed the ugly fancy

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Fate of Biological Contaminants During

Recycling of Organic Wastes - eBook

Copyright © 2023 Elsevier Inc. All rights reserved.

For information on all Elsevier publications

Publisher: Joseph P. Hayton

1. Occurrence, fate, and behavior of antibiotic resistance genes

2. Human pathogenic microorganism in sludge 27

3. Potential of entomocomposting toward soil pathogen

4. Valorization of fish wastes for circular bioeconomy:

5. Dynamics of biological contaminants along with microbial

3. Composition and diversity of microbes associated with

6. Treatment approaches for bio-contaminants in organic

7. Fate of bio-contaminants in organic wastes during

8. Metagenomic analysis revealing the dual microbial community

9. Solid waste management during the COVID-19 pandemic 177

10. Effects of bio-contaminants in organic waste products

11. Fate of bio-contaminants in soil systems and available

2. Occurrence of bio-contaminants from manure, sludge,

12. Antibiotics and antibiotics resistance genes in poultry

13. Vermicomposting as a means of removing antibiotic

14. Insight on the prevalence of pathogens present in the municipal

2. Sources, transmission route, and fate of pathogens in wastes and

15. Bioconversion of hazardous organic wastes using

16. Effects of plants’ density on the reduction of antibiotic

Ganesh Kumar Agrawal

Sartaj Ahmad Bhat

Susanta Kumar Chakraborty

Saikat Ranjan Das

Daljeet Singh Dhanjal

Ram Kumar Ganguly

Vaibhav Vasant Goud

Bitish Kumar Nayak

Angie M. Pedraza Torres

Chrysantus Mbi Tanga

Occurrence, fate, and behavior

Fate of Biological Contaminants During Recycling 1

clarification of the occurrence and behavior of antibiotics, ARGs and MGEs

Fig. 1 Speading pathways of antibiotics, antibiotic resistance bacteria (ARB) harboring

2. Occurrence of antibiotics, ARGs, and MGEs in various

Fig. 2 Main mechanisms for bacteria to be antibiotic-resistant.

the usage of large quantities of tetracycline in clinics, livestock, and poultry

3. Fate and behavior of ARGs and MGEs during different

issue caused by newly emerging contaminants. Table 2 exhibited the results

Treatments Organic wastes Conditions Additives Type Before After Type

Dewatered sludge Mesophilic (35 1°C); Inoculum sludge from a / / / tetA,

Swine manure 37 0.1°C; 30 days Inoculum sludge; / / / 12 ARGs

Dairy manure Mesophilic (37°C) and / / / / tetA, tetC,

(1) Aerobic Food waste (1) Inoculum sludge 10 ARGs

Before After Type Before After Main findings References

(1) 2.4 1012–2.5 1016; (1) 1.7 1010–3.3 1012;

Treatments Organic wastes Conditions Additives Type Before After Type

Mixture of swine 43 days (55°C for 7 days) / / / / aadA,

(1) Fruit and 10 earthworms/ 100 g; / / / / tetG,

(1), (2), (3) and (4): different treatments. /: No data.

Before After Type Before After Main findings References

during composting of swine manure described that the maturity period

7. Elimination strategies for ARGs and MGEs during

8. Conclusion and perspectives

product to the soil environment or the vegetation needs to be clarified. Fur-

of dewatered sludge. Environ. Pollut. 252, 1068–1077. https://doi.org/10.1016/j.

Fate of Biological Contaminants During Recycling 27

Fig. 1 Origin of primary and secondary sludge in a sewage treatment sequence.

by-products are generated. The sludge generated during the wastewater

physical systems. The stabilized sludge is called a biosolid. Once stabilized,