Science of the Total Environment 654 (2019) 452–455

Contents lists available at ScienceDirect

Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

Short Communication

Antibiotic resistance genes in bacteriophages from diverse

marine habitats
William Calero-Cáceres a, José Luis Balcázar b,⁎
a
UTA-RAM-OneHealth Group, Centro de Investigaciones Agropecuarias, Facultad de Ciencias Agropecuarias, Universidad Técnica de Ambato, Tungurahua, Ecuador
b
Catalan Institute for Water Research (ICRA), University of Girona, Girona, Spain

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Phages from marine habitats are poten-

tial reservoirs of antibiotic resistance
genes (ARGs).
• Several ARGs were found in all analyzed
marine viromes.
• The role of phages as reservoirs of ARGs
should not be underestimated.

a r t i c l e i n f o a b s t r a c t

Article history: Although antibiotic resistance represents a significant and growing threat to human and environmental health
Received 3 September 2018 worldwide, the contribution of bacteriophages (phages) to the acquisition and spread of antibiotic resistance
Received in revised form 18 October 2018 genes (ARGs) in the environment has not been extensively explored. In this study, a comprehensive analysis of
Accepted 11 November 2018
several viromes from diverse marine habitats was performed to investigate whether or not phages carry ARGs.
Available online 12 November 2018
The analysis provides strong evidence that phages from marine habitats are potential reservoirs of ARGs. In
Editor: Ewa Korzeniewska fact, genes conferring resistance to aminocoumarin, bacitracin and multidrug resistance (particularly the mexB
gene) were found in all analyzed marine viromes. Given this, the role of phages as reservoirs of ARGs should
Keywords: not be underestimated considering their global distribution.
Marine bacteriophages © 2018 Elsevier B.V. All rights reserved.
Environmental reservoirs
Antibiotic resistance genes

1. Introduction Health Organization 2015). The current trend in antimicrobial resis-

In the last century, anthropogenic activities have exerted a strong
evolutionary pressure, causing a disturbance in the microbiome, and
thus in the resistome (Zhu et al. 2017). In fact, the indiscriminate use
of antimicrobial agents has led to the emergence and selection of resis-
tant bacteria, which has become a major global health threat (World
⁎ Corresponding author.
E-mail address: jlbalcazar@icra.cat (J.L. Balcázar).
tance is expecting a significant increase in mortality worldwide unless
a global mitigation strategy is applied (de Kraker et al. 2016; O'Neill
2014). A sustainable action should therefore include the links among
human, animal and environmental health, according to the One Health
approach (Queenan et al. 2016; Robinson et al. 2016).
A growing number of studies are demonstrating the presence of an-
timicrobial resistance hotspots in different environments and their po-
tential influence in the evolution and spread of antibiotic resistance
genes (ARGs) (Huijbers et al. 2015; Surette and Wright 2017). Among

https://doi.org/10.1016/j.scitotenv.2018.11.166
0048-9697/© 2018 Elsevier B.V. All rights reserved.
 W. Calero-Cáceres, J.L. Balcázar / Science of the Total Environment 654 (2019) 452–455
them, aquatic ecosystems represent one of the most suitable reactors for
the emergence and spread of these ARGs (Marti et al. 2014a). Different
mobile genetic elements carrying ARGs have been identified such as in-
sertion sequences, transposons, integrative conjugative elements, plas-
mids, gene transfer agents and bacteriophages (phages) (Brown-Jaque
et al. 2015; Lekunberri et al. 2017). Phages are the most abundant bio-
logical entities in the biosphere, and they have the potential to transfer
genetic material via transduction between bacterial hosts (Balcazar
2014; Sime-Ngando 2014).
Phages have emerged as prime suspects in bacterial adaptation and
evolution, thereby making them suitable vehicles for acquisition, main-
tenance and spread of ARGs (Koskella and Brockhurst 2014). Recent
studies have demonstrated that phages carrying ARGs are widely dis-
tributed in different environments, particularly those impacted by an-
thropogenic activities (Calero-Cáceres et al. 2014; Colomer-Lluch et al.
2014a; Lekunberri et al. 2017; Marti et al. 2014b; Subirats et al. 2016).
Moreover, phage particles carrying ARGs have shown a greater persis-
tence than those ARGs packaged into bacterial cells during natural inac-
tivation and common disinfection processes (Calero-Cáceres and
Muniesa 2016). Its stability poses a threat because the possibility of hor-
izontal gene transfer events could increase in natural environments.
Despite cutting-edge genomic technologies have recently led to sig-
nificant advances in our understanding of the role of phages in mobiliz-
ing ARGs (Colombo et al. 2017; Kim et al. 2017), their contribution to
antibiotic resistance in marine habitats has not been fully explored.
Here, a comprehensive analysis of several viromes from diverse marine
habitats was performed to investigate whether or not phages carry
ARGs and, if so, the diversity and abundance of these genes.
2. Materials and methods
Seventeen viromes from diverse marine habitats were obtained
from public repositories (Supplementary Table S1). Low-quality reads
from each virome were initially filtered using the FASTX-Toolkit
(http://hannonlab.cshl.edu/fastx_toolkit/) with a quality cut-off of 20.
High-quality reads were then screened for the presence of 16S rRNA
gene sequences using METAXA2 (Bengtsson-Palme et al. 2015), as the
abundance of ARGs in viromes can be overestimated due to bacterial
DNA contamination (Enault et al. 2017). Results showed that all se-
lected viromes had no or low levels of bacterial DNA (b0.2‰ of 16S
rRNA gene reads), thus validating subsequent analyses. BLASTX imple-
mented in the DIAMOND software (Buchfink et al. 2015) was then
employed to align reads of each virome against the DeepARG database
Fig. 1. Relative abundance of antibiotic resistance genes (ARGs) in viromes from different marine habitats, which are distributed according to the class of antibiotics that they confer resistance to.
Counts were normalized to the total number of reads per virome. MLS, macrolide-lincosamide-streptogramin B. Multidrug indicates resistance to at least three classes of antibiotics.
453
(Arango-Argoty et al. 2018). A read was annotated as a resistance
gene if the best BLAST hit (BLASTX) had ≥80% amino acid identity over
at least 25 amino acids of the target sequence. This stringent threshold
was chosen to minimize false positives (Lekunberri et al. 2017). The
abundance of ARGs was normalized to the total number of reads per
virome, which were grouped according to the class of antibiotics that
they confer resistance to. Moreover, diversity of ARGs was calculated
as previously described (Pal et al. 2016).
3. Results and discussion
The role of phages in marine ecosystems is highly important, be-
cause they can regulate bacterial densities, influencing elemental cy-
cling (Jover et al. 2014). Moreover, there is strong evidence that
supports the mutual evolution between phages and their bacterial
hosts (Argov et al. 2017; Haaber et al. 2016). They can also influence
horizontal gene transfer between taxa by transduction, conferring evo-
lutionary advantages to microorganisms (Williams 2013). One of the
most important traits that phages can confer to their bacterial hosts is
antibiotic resistance determinants (Keen and Dantas 2018). In vivo
studies suggest that antibiotic perturbation stimulates the phage-
packaged ARGs in bacteria (Modi et al. 2013). Considering the occur-
rence and persistence of antibiotics in superficial water (particularly in
sewage-impacted sites), including seawater (Gothwal and Shashidhar
2015), it is possible that their residues may trigger ecological interac-
tions between phages and their host, thereby promoting the phage-
packaged ARGs under such challenging conditions (e.g., anthropogenic
impact).
In this study, we report the presence of ARGs in different marine
viromes around the world. It is important to consider the potential
biases related with sequencing technologies, nucleic acid extraction
methods and library preparation that could influence the diversity and
relative abundance in samples from environments with low occurrence
of ARGs (Brooks et al. 2015; Parras-Moltó et al. 2018). To overcome
these potential biases, unassembled reads were used to make quantita-
tive comparisons among the selected viromes. This analysis showed
that most marine viromes harbored relatively few ARGs (ranging from
0.004% in the Bay of British Columbia to 0.05% in the South Sea of
Korea), except in the Indian Ocean viromes, which reached a high abun-
dance (from 0.08% to 0.22%) (Fig. 1). Genes conferring resistance to
aminocoumarin (inhibitor of bacterial DNA gyrase), bacitracin (poly-
peptide that inhibits peptidoglycan synthesis) and multidrug resistance
(resistance to at least three classes of antibiotics) were found in all
454 W. Calero-Cáceres, J.L. Balcázar / Science of the Total Environment 654 (2019) 452–455
analyzed marine viromes. Interestingly, the mexB gene, belonging to the
resistance-nodulation-cell division (RND) family efflux pump, was
found to be prevalent among the genes conferring multidrug resistance.
Although direct comparisons with other studies cannot be
established due to the origin of samples, these results seem to be consis-
tent with previous studies, which have shown that viromes from other
habitats (e.g., anthropogenically impacted and non-impacted freshwa-
ter environments) are enriched for genes encoding antibiotic resistance
(Colombo et al. 2017; Lekunberri et al. 2017). Genes conferring multi-
drug and aminocoumarin resistance have also been found in
metagenomes from diverse environments, such as pristine arctic wet-
land (Diaz et al. 2017), flood-impacted river (Garner et al. 2016) or sed-
iments (Chu et al. 2018), whereas bacitracin resistance genes have
commonly been detected in human gut metagenomes (Hu et al. 2013;
Feng et al. 2018).
Moreover, comparative analysis revealed marked differences in the
diversity and abundance of ARGs according to their origin (Supplemen-
tary Fig. 1). Viromes from the South Sea of Korea had the highest diver-
sity of ARGs, whereas the Indian Ocean viromes were dominated by
genes conferring resistance to β-lactams, tetracyclines and multidrug
resistance. According with the sample site maps, samples from Korea
were taken from sites near coastland (Hwang et al. 2017), whereas sam-
ples from Indian Ocean were collected from interior ocean waters, with
a considerable distance from continents (Williamson et al. 2012). It ap-
pears that the diversity of ARGs in South Sea of Korea locations could be
influenced by anthropogenic activities more than Indian Ocean samples.
Fluoroquinolone resistance genes, whose origin has been associated to
aquatic microorganisms (Poirel et al. 2005), were detected in all the
samples from South Sea of Korea locations. This confirms previously
published findings, indicating that the detection of phage-related fluo-
roquinolone resistance genes is common in anthropogenically polluted
environments (Colomer-Lluch et al. 2014b; Marti et al. 2014b). Al-
though the diversity of ARGs was relatively low in Indian Ocean
viromes, their abundance was high, particularly those genes conferring
resistance to β-lactams (particularly blaTEM genes) and tetracyclines
(particularly tetC genes). These genes have been reported to be widely
distributed, including in environmental settings (Anand et al. 2016;
Calero-Cáceres et al. 2017; Colomer-Lluch et al. 2014a). The blaTEM
genes confer resistance to penicillin family antibiotics such as ampicillin
and oxacillin, and recent studies have suggested them as indicator genes
of antibiotic resistance pollution in anthropogenically impacted envi-
ronments, considering their abundance and dissemination patterns in
different environments (Devarajan et al. 2016; Narciso-Da-Rocha et al.
2014; Zhu et al. 2017). However, their presence should not be consid-
ered as a defining feature of anthropogenic pollution, because a wide di-
versity of ARGs has also been detected in pristine environments
(D'Costa et al. 2011).
While a clear association between phages carrying ARGs and pol-
luted environments has not yet been established, phages may provide
their bacterial hosts with a substantial advantage under challenging
conditions. These results therefore suggest that the Indian Ocean is sub-
jected to a variety of pollution sources that would favor the acquisition
and spread of ARGs via phages, although these results should be
interpreted with caution due to the number of analyzed viromes.
4. Conclusion
In conclusion, phages play a critical role in the microbial ecology of
many habitats due to their high abundance and diversity; however, lim-
ited information is available on their contribution to antibiotic resis-
tance. These results thus provide strong evidence that phages from
marine habitats are potential reservoirs of ARGs. Considering the global
distribution of phages, their role in the acquisition and spread of ARGs
should not be underestimated. In fact, these aspects should be taken
into account in order to provide new avenues to tackle the global crisis
of antibiotic resistance.
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2018.11.166.
Funding information
This study was supported by the Economy and Knowledge Depart-
ment of the Catalan Government through Consolidated Research
Group (ICRA-ENV 2017 SGR 1124).
Compliance with ethical standards
Not required.
References
Anand, T., Bera, B.C., Vaid, R.K., Barua, S., Riyesh, T., Virmani, N., Hussain, M., Singh, R.K.,
Tripathi, B.N., 2016. Abundance of antibiotic resistance genes in environmental bacte-
riophages. J. Gen. Virol. 97, 3458–3466. https://doi.org/10.1099/jgv.0.000639.
Arango-Argoty, G., Garner, E., Pruden, A., Heath, L.S., Vikesland, P., Zhang, L., 2018.
DeepARG: a deep learning approach for predicting antibiotic resistance genes from
metagenomic data. Microbiome 6, 23. https://doi.org/10.1186/s40168-018-0401-z.
Argov, T., Azulay, G., Pasechnek, A., Stadnyuk, O., Ran-Sapir, S., Borovok, I., Sigal, N.,
Herskovits, A.A., 2017. Temperate bacteriophages as regulators of host behavior.
Curr. Opin. Microbiol. 38, 81–87. https://doi.org/10.1016/j.mib.2017.05.002.
Balcazar, J.L., 2014. Bacteriophages as vehicles for antibiotic resistance genes in the envi-
ronment. PLoS Pathog. 10, e1004219. https://doi.org/10.1371/journal.ppat.1004219.
Bengtsson-Palme, J., Hartmann, M., Eriksson, K.M., Pal, C., Thorell, K., Larsson, D.G.,
Nilsson, R.H., 2015. METAXA2: improved identification and taxonomic classification
of small and large subunit rRNA in metagenomic data. Mol. Ecol. Resour. 15,
1403–1414. https://doi.org/10.1111/1755-0998.12399.
Brooks, J.P., Edwards, D.J., Harwich, M.D., Rivera, M.C., Fettweis, J.M., Serrano, M.G., Reris,
R.A., Sheth, N.U., Huang, B., Girerd, P., Strauss, J.F., Jefferson, K.K., Buck, G.A., Buck, G.A.,
2015. The truth about metagenomics: quantifying and counteracting bias in 16S
rRNA studies. BMC Microbiol. 15, 66. https://doi.org/10.1186/s12866-015-0351-6.
Brown-Jaque, M., Calero-Cáceres, W., Muniesa, M., 2015. Transfer of antibiotic-resistance
genes via phage-related mobile elements. Plasmid 79, 1–7. https://doi.org/10.1016/j.
plasmid.2015.01.001.
Buchfink, B., Xie, C., Huson, D.H., 2015. Fast and sensitive protein alignment using DIA-
MOND. Nat. Methods 12, 59–60. https://doi.org/10.1038/nmeth.3176.
Calero-Cáceres, W., Muniesa, M., 2016. Persistence of naturally occurring antibiotic resis-
tance genes in the bacteria and bacteriophage fractions of wastewater. Water Res. 95,
11–18. https://doi.org/10.1016/j.watres.2016.03.006.
Calero-Cáceres, W., Melgarejo, A., Colomer-Lluch, M., Stoll, C., Lucena, F., Jofre, J., Muniesa,
M., 2014. Sludge as a potential important source of antibiotic resistance genes in both
the bacterial and bacteriophage fractions. Environ. Sci. Technol. 48, 7602–7611.
https://doi.org/10.1021/es501851s.
Calero-Cáceres, W., Méndez, J., Martín-Díaz, J., Muniesa, M., 2017. The occurrence of anti-
biotic resistance genes in a Mediterranean river and their persistence in the riverbed
sediment. Environ. Pollut. 223, 384–394. https://doi.org/10.1016/j.
envpol.2017.01.035.
Chu, B.T.T., Petrovich, M.L., Chaudhary, A., Wright, D., Murphy, B., Wells, G., Poretsky, R.,
2018. Metagenomic analysis reveals the impact of wastewater treatment plants on
the dispersal of microorganisms. Appl. Environ. Microbiol. 84. https://doi.org/
10.1128/AEM.02168-17 (e02168-17).
Colombo, S., Arioli, S., Neri, E., Della Scala, G., Gargari, G., Mora, D., 2017. Viromes as ge-
netic reservoir for the microbial communities in aquatic environments: a focus on
antimicrobial-resistance genes. Front. Microbiol. 8, 1–13. https://doi.org/10.3389/
fmicb.2017.01095.
Colomer-Lluch, M., Calero-Cáceres, W., Jebri, S., Hmaied, F., Muniesa, M., Jofre, J., 2014a.
Antibiotic resistance genes in bacterial and bacteriophage fractions of Tunisian and
Spanish wastewaters as markers to compare the antibiotic resistance patterns in
each population. Environ. Int. 73, 167–175. https://doi.org/10.1016/j.
envint.2014.07.003.
Colomer-Lluch, M., Jofre, J., Muniesa, M., 2014b. Quinolone resistance genes (qnrA and
qnrS) in bacteriophage particles from wastewater samples and the effect of inducing
agents on packaged antibiotic resistance genes. J. Antimicrob. Chemother. 69,
1265–1274. https://doi.org/10.1093/jac/dkt528.
D'Costa, V.M., King, C.E., Kalan, L., Morar, M., Sung, W.W.L., Schwarz, C., Froese, D., Zazula,
G., Calmels, F., Debruyne, R., Golding, G.B., Poinar, H.N., Wright, G.D., 2011. Antibiotic
resistance is ancient. Nature 477, 457–461. https://doi.org/10.1038/nature10388.
Devarajan, N., Laffite, A., Mulaji, C.K., Otamonga, J.P., Mpiana, P.T., Mubedi, J.I., Prabakar, K.,
Ibelings, B.W., Poté, J., 2016. Occurrence of antibiotic resistance genes and bacterial
markers in a tropical river receiving hospital and urban wastewaters. PLoS One 11,
e0149211. https://doi.org/10.1371/journal.pone.0149211.
Diaz, K.S., Rich, V.I., McLain, J.E., 2017. Searching for antibiotic resistance genes in a pris-
tine Arctic wetland. J. Contemp. Water Res. Educ. 160, 42–59. https://doi.org/
10.1111/j.1936-704X.2017.03239.x.
Enault, F., Briet, A., Bouteille, L., Roux, S., Sullivan, M.B., Petit, M.A., 2017. Phages rarely en-
code antibiotic resistance genes: a cautionary tale for virome analyses. ISME J. 11,
237–247. https://doi.org/10.1038/ismej.2016.90.
 W. Calero-Cáceres, J.L. Balcázar / Science of the Total Environment 654 (2019) 452–455
Feng, J., Li, B., Jiang, X., Yang, Y., Wells, G.F., Zhang, T., Li, X., 2018. Antibiotic resistome in a
large-scale healthy human gut microbiota deciphered by metagenomic and network
analyses. Environ. Microbiol. 20, 355–368. https://doi.org/10.1111/1462-2920.14009.
Garner, E., Wallace, J.S., Argoty, G.A., Wilkinson, C., Fahrenfeld, N., Heath, L.S., Zhang, L.,
Arabi, M., Aga, D.S., Pruden, A., 2016. Metagenomic profiling of historic Colorado
Front Range flood impact on distribution of riverine antibiotic resistance genes. Sci.
Rep. 6, 38432. https://doi.org/10.1038/srep38432.
Gothwal, R., Shashidhar, T., 2015. Antibiotic pollution in the environment: a review.
Clean: Soil, Air, Water 43, 479–489. https://doi.org/10.1002/clen.201300989.
Haaber, J., Leisner, J.J., Cohn, M.T., Catalan-Moreno, A., Nielsen, J.B., Westh, H., Penadés, J.R.,
Ingmer, H., 2016. Bacterial viruses enable their host to acquire antibiotic resistance
genes from neighbouring cells. Nat. Commun. 7, 13333. https://doi.org/10.1038/
ncomms13333.
Hu, Y., Yang, X., Qin, J., Lu, N., Cheng, G., Wu, N., Pan, Y., Li, J., Zhu, L., Wang, X., Meng, Z.,
Zhao, F., Liu, D., Ma, J., Qin, N., Xiang, C., Xiao, Y., Li, L., Yang, H., Wang, J., Yang, R.,
Gao, G.F., Wang, J., Zhu, B., 2013. Metagenome-wide analysis of antibiotic resistance
genes in a large cohort of human gut microbiota. Nat. Commun. 4, 2151. https://
doi.org/10.1038/ncomms3151.
Huijbers, P.M.C., Blaak, H., De Jong, M.C.M., Graat, E.A.M., Vandenbroucke-Grauls, C.M.J.E.,
De Roda Husman, A.M., 2015. Role of the environment in the transmission of antimi-
crobial resistance to humans: a review. Environ. Sci. Technol. 49, 11993–12004.
https://doi.org/10.1021/acs.est.5b02566.
Hwang, J., Park, S.Y., Park, M., Lee, S., Lee, T.K., 2017. Seasonal dynamics and metagenomic
characterization of marine viruses in Goseong Bay, Korea. PLoS One 12, e0169841.
https://doi.org/10.1371/journal.pone.0169841.
Jover, L.F., Effler, T.C., Buchan, A., Wilhelm, S.W., Weitz, J.S., 2014. The elemental composi-
tion of virus particles: implications for marine biogeochemical cycles. Nat. Rev.
Microbiol. 12, 519–528. https://doi.org/10.1038/nrmicro3289.
Keen, E.C., Dantas, G., 2018. Close encounters of three kinds: bacteriophages, commensal
bacteria, and host immunity. Trends Microbiol. 26, 943–954. https://doi.org/10.1016/
j.tim.2018.05.009.
Kim, Y., Van Bonn, W.V., Aw, T.G., Rose, J.B., 2017. Aquarium viromes: Viromes of human-
managed aquatic systems. Front. Microbiol. 8, 1231. https://doi.org/10.3389/
fmicb.2017.01231.
Koskella, B., Brockhurst, M.A., 2014. Bacteria-phage coevolution as a driver of ecological
and evolutionary processes in microbial communities. FEMS Microbiol. Rev. 38,
916–931. https://doi.org/10.1111/1574-6976.12072.
de Kraker, M.E.A., Stewardson, A.J., Harbarth, S., 2016. Will 10 million people die a year
due to antimicrobial resistance by 2050? PLoS Med. 13, e1002184. https://doi.org/
10.1371/journal.pmed.1002184.
Lekunberri, I., Subirats, J., Borrego, C.M., Balcázar, J.L., 2017. Exploring the contribution of
bacteriophages to antibiotic resistance. Environ. Pollut. 220, 981–984. https://doi.org/
10.1016/j.envpol.2016.11.059.
Marti, E., Variatza, E., Balcázar, J.L., 2014a. The role of aquatic ecosystems as reservoirs of
antibiotic resistance. Trends Microbiol. 22, 36–41. https://doi.org/10.1016/j.
tim.2013.11.001.
Marti, E., Variatza, E., Balcázar, J.L., 2014b. Bacteriophages as a reservoir of extended-
spectrum β-lactamase and fluoroquinolone resistance genes in the environment.
Clin. Microbiol. Infect. 20, O456–O459. https://doi.org/10.1111/1469-0691.12446.
455
Modi, S.R., Lee, H.H., Spina, C.S., Collins, J.J., 2013. Antibiotic treatment expands the resis-
tance reservoir and ecological network of the phage metagenome. Nature 499,
219–222. https://doi.org/10.1038/nature12212.
Narciso-Da-Rocha, C., Varela, A.R., Schwartz, T., Nunes, O.C., Manaia, C.M., 2014. blaTEM
and vanA as indicator genes of antibiotic resistance contamination in a hospital-
urban wastewater treatment plant system. J. Glob. Antimicrob. Resist. 2, 309–315.
https://doi.org/10.1016/j.jgar.2014.10.001.
O'Neill, J., 2014. Antimicrobial Resistance: Tackling a crisis for the health and wealth of na-
tions. https://amr-review.org/Publications.html, Accessed date: 3 September 2018.
Pal, C., Bengtsson-Palme, J., Kristiansson, E., Larsson, D.G.J., 2016. The structure and diver-
sity of human, animal and environmental resistomes. Microbiome 4, 54. https://doi.
org/10.1186/s40168-016-0199-5.
Parras-Moltó, M., Rodríguez-Galet, A., Suárez-Rodríguez, P., López-Bueno, A., 2018. Evalu-
ation of bias induced by viral enrichment and random amplification protocols in
metagenomic surveys of saliva DNA viruses. Microbiome 6, 119. https://doi.org/
10.1186/s40168-018-0507-3.
Poirel, L., Liard, A., Nordmann, P., Mammeri, H., 2005. Origin of plasmid-mediated quino-
lone resistance determinant QnrA. Antimicrob. Agents Chemother. 49, 3523–3525.
https://doi.org/10.1128/AAC.49.8.3523.
Queenan, K., Hasler, B., Rushton, J., 2016. A one health approach to antimicrobial resis-
tance surveillance: is there a business case for it? Int. J. Antimicrob. Agents 48,
422–427. https://doi.org/10.1016/j.ijantimicag.2016.06.014.
Robinson, T.P., Bu, D.P., Carrique-Mas, J., Fèvre, E.M., Gilbert, M., Grace, D., Hay, S.I.,
Jiwakanon, J., Kakkar, M., Kariuki, S., Laxminarayan, R., Lubroth, J., Magnusson, U.,
Thi Ngoc, P., Van Boeckel, T.P., Woolhouse, M.E.J., 2016. Antibiotic resistance is the
quintessential one health issue. Trans. R. Soc. Trop. Med. Hyg. 110, 377–380.
https://doi.org/10.1093/trstmh/trw048.
Sime-Ngando, T., 2014. Environmental bacteriophages: viruses of microbes in aquatic
ecosystems. Front. Microbiol. 5, 355. https://doi.org/10.3389/fmicb.2014.00355.
Subirats, J., Sànchez-Melsió, A., Borrego, C.M., Balcázar, J.L., Simonet, P., 2016.
Metagenomic analysis reveals that bacteriophages are reservoirs of antibiotic resis-
tance genes. Int. J. Antimicrob. Agents 48, 163–167. https://doi.org/10.1016/j.
ijantimicag.2016.04.028.
Surette, M., Wright, G.D., 2017. Lessons from the environmental antibiotic resistome.
Annu. Rev. Microbiol. 71, 309–329. https://doi.org/10.1146/annurev-micro-090816-
093420.
Williams, H.T.P., 2013. Phage-induced diversification improves host evolvability. BMC
Evol. Biol. 13, 17. https://doi.org/10.1186/1471-2148-13-17.
Williamson, S.J., Allen, L.Z., Lorenzi, H.A., Fadrosh, D.W., Brami, D., Thiagarajan, M.,
McCrow, J.P., Tovchigrechko, A., Yooseph, S., Venter, J.C., 2012. Metagenomic explora-
tion of viruses throughout the Indian Ocean. PLoS One 7, e42047. https://doi.org/
10.1371/journal.pone.0042047.
World Health Organization, 2015. Antimicrobial resistance [WWW Document]. Fact sheet
N°194. http://www.who.int/mediacentre/news/releases/2014/amr-report/en/,
Accessed date: 17 March 2016.
Zhu, Y.G., Gillings, M., Simonet, P., Stekel, D., Banwart, S., Penuelas, J., 2017. Microbial mass
movements. Science 357, 1099–1100. https://doi.org/10.1126/science.aao3007.