Ann Microbiol
DOI 10.1007/s13213-014-0911-2
REVIEW ARTICLE
The prevalence of antibiotic resistance genes among Aeromonas
species in aquatic environments
Marta Piotrowska & Magdalena Popowska
Received: 10 January 2014 / Accepted: 5 May 2014
# Springer-Verlag Berlin Heidelberg and the University of Milan 2014
Abstract The global rise in antimicrobial resistance (AMR)
among bacteria causing infectious diseases is well documented,
and the associated risks for human health are well known. There
is much less research on AMR with regard to environmental
strains, both opportunistic and pathogenic ones. The genus
Aeromonas is widely distributed in the environment and causes
many variable diseases in fish and humans. Infections in humans
are predominantly caused by Aeromonas veronii, A. hydrophila
and A. caviae (A. punctata) in a form of bacteremia, gastroen-
teritis or even septicaemia in immunocompetent and immuno-
compromised individuals. Different groups of antibiotics are
used in the treatment, but studies indicate that fluoroquinolones
and cefotaxime are the most efficient. A disturbing consequence
of antibiotic overuse is an increasing number of detection of
various antibiotic resistance genes (ARG) within this genus. The
water environment is one of the major modes of transmission of
resistant bacteria from animals to humans, and, thus, the dis-
semination of antibiotic resistance genes, particularly those
located in mobile genetic elements (MGE) occurs in such as
plasmids and transposons. This review summarizes recently
published information on the type, distribution, and transmission
of ARG by MGE, widespread in Aeromonas strains living in
various aquatic environments, including wastewater, natural
water, aquaculture and urban drinking water. The data available
indicate that the opportunistic pathogens like Aeromonas spp.
might serve as important vectors of ARG for clinically relevant
pathogens present in such bodies of water .
M. Piotrowska : M. Popowska
Department of Applied Microbiology, Institute of Microbiology,
Faculty of Biology, University of Warsaw, Miecznikowa 1,
02-096 Warsaw, Poland
M. Piotrowska (*)
Institute of Microbiology, Faculty of Biology, University of Warsaw,
Miecznikowa 1, 02-096 Warsaw, Poland
e-mail: m.piotrowska@biol.uw.edu.pl
Keywords Aeromonas . Antibiotic resistance genes . Natural
water . Aquacultures . Urban drinking water . Wastewater
treatment plant
Introduction
Species belonging to the genus Aeromonas are widely distrib-
uted in the environment. They are able to inhabit natural soil,
food and animals, but they most commonly occur in all kinds
of aquatic environments (Janda and Abott 2010). Aeromonas
spp. have been isolated from wastewater (Figueira et al. 2011),
natural water such as rivers, lakes and estuaries (Henriques
et al. 2006; Picão et al. 2008), aquacultures (Schmidt et al.
2001; Sørum 2006) and urban drinking water (Carvalho et al.
2012). The genus Aeromonas has a complex taxonomy at the
species level, but it belongs to the family Aeromondaceae,
which is part of the class of Gamma-proteobacteria. The
species classification changes dynamically with the emer-
gence of new genetic analyses, but thus far 28 species have
been validated or proposed (http://www.bacterio.net).
Aeromonas species belong to one of the two major groups:
psychrophilic or mesophilic bacteria. The former represents
nonmotile bacteria with optimal growth temperature balanced
between 22 - 25 °C that cause many variable diseases in fish,
e.g., furunculosis, septicaemia, ulcerative or hemorrhagic dis-
eases (Beaz-Hidalgo and Figueras 2013; Dallaire-Dufresne
et al. 2013). In contrast, mesophilic species are motile, their
optimal growth temperature is between 35 - 37 °C, and being
opportunistic pathogens of humans, they are potentially dan-
gerous. Infections are mostly induced by A. veronii, A.
hydrophila and A. caviae (A. punctata) and result in bacter-
emia, gastroenteritis or even septicaemia in immunocompe-
tent and immunocompromised individuals (Lai et al. 2007;
Parker and Shaw 2010). The number of infectious cases
increases in the warmer months and the most common

manifestations are diarrhea and skin infections after contact
with contaminated water. Treatment of these infections varies
and depends on the part of the world, but studies indicate that
fluoroquinolones and cefotaxime are the most successful ther-
apies (Parker and Shaw 2010). Aeromonas infections are not
an important public health problem so epidemiology is not
very well known. However, previous studies carried out in
different countries suggested that these infections should not
be underestimated (Ghenghesh et al. 2008). Given the fact that
Aeromonas spp. are opportunistic pathogens of humans and
the causative agent of fish diseases, it is disturbing that more
and more different antibiotic resistance genes (ARG) are being
detected within this genus (Zhang et al. 2009a). The main
targets of Aeromonas are fish, which are exposed to these
natural pathogens. To study the prevalence and horizontal
transfer of resistance genes within this genus, it is important
to examine the aquatic environment as a whole, including fish
and sediments.
ARG are ubiquitous both in clinical pathogens and envi-
ronmental bacterial species. This is in part due to the wide
usage of antibiotics in clinical practice, veterinary medicine
and agriculture (Cantas et al. 2013), resulting in the release of
large amounts of these pollutants to the environment. Antibi-
otics, in addition to being chemical pollutants, exert a selective
pressure retaining and spreading the ARG among microbiota,
which poses a risk to human health (Kemper 2008). Addition-
ally, it has been demonstrated that heavy metals can co-select
the spread of ARG in the environment (Seiler and Berendonk
2012). Consequently, natural environments, especially water
bodies, constitute important reservoirs of ARG that may pro-
mote the spread of resistance to different groups of antibiotics
across different bacterial species (Lupo et al. 2012). Hence,
opportunistic pathogens such as Aeromonas spp. might be
dangerous vectors of ARG for clinically relevant pathogens,
which also occur in the water (Rhodes et al. 2000; Sørum
2006).
In this review we strive to present the latest information
from the literature on the prevalence of ARG in Aeromonas
spp. from various water environments endowed with different
anthropogenic impact. The main focus is on the differences
between natural water, drinking water and more polluted
environments such as wastewater and aquacultures. Further,
our intent is to highlight the problem of the horizontal transfer
of these genes, which is related to the presence of integrons
and mobile genetic elements such as plasmids, transposons or
insertion sequences, and the serious risks for human health
related to this process.
Wastewater treatment plants
Wastewater treatment plants (WWTP) constitute a specific
kind of water environment because of the prevailing good
Ann Microbiol
conditions for the proliferation of bacteria (richness of nutri-
ents, aeration, optimal temperature). Particularly favorable
conditions exist in the activated sludge, which is characterized
by a high density and variety of microorganisms. Moreover,
WWTP receive large amounts of antibiotics with sewage
water from hospitals, livestock manure and private house-
holds, where bacteria are exposed to selection pressure
(Bouki et al. 2013). These conditions promote the prolifera-
tion of antibiotic resistant bacteria (ARB) and increase the
probability of the horizontal gene transfer (HGT) of ARG
(Tennstedt et al. 2003).
The study on sewage samples using high-throughput se-
quencing of 16S rRNA gene has detected over ten bacterial
phyla with the highest contribution being from Proteobacteria,
Actinobactera, Chloroflexi, Firmicutes, Bacteriodetes and
Nitrospirae (Ye et al. 2012) as well as a large number of
Aeromonas spp. (Moura et al. 2007; Figueira et al. 2011;
Igbinosa and Okoh 2012; Moura et al. 2012; Igbinosa and
Okoh 2013). Treatment processes reduce bacterial count in the
sewage, including the ARB (Guardabassi et al. 2002). How-
ever, even in effluent water, ARG can be taken up by trans-
formation and ARB are still detected (Zhang et al. 2009a).
In the literature there is little information about ARG in
Aeromonas spp. present in WWTP. A variety of different
species that has been detected so far include
A. allosaccharophila, A. aquariorum, A. hydrophila, A. me-
dia, A. veronii, A. caviae, A. sanarellii and A. taiwanesis,
(Figueira et al. 2011) (Table 1). The ARG recently found in
Aeromonas strains from wastewater encoded resistance to four
major groups of antibiotics, i.e., the quinolones, aminoglyco-
sides, β-lactams and tetracyclines. In contrast to previous
studies, Figueira et al. (2011) who isolated bacteria from
municipal sewage, argued that the vast majority of quinolone
resistance was the result of chromosomal mutations. These
authors found many mutations in gyrA and parC genes, which
are genetic determinants of quinolone resistance. The majority
of these strains was resistant to nalidixic acid and belonged to
A. media and A. caviae species. This high frequency of muta-
tions could be the effect of contact with a high concentration
of mutagenic substances in wastewater (Miyahara et al. 2011).
These authors also found two plasmid-mediated quinolone
resistance genes: qnrS and accA-cr, but only in A. media
strains. In addition, this study investigated the occurrence of
the cphA chromosomal gene, which is the most common
metallo-β-lactamase in Aeromonas spp. (Janda and Abott
2010). This gene was detected in 18 % of strains in WWTP.
The results of the study by Balsalobre et al. (2009) indicated
that the cphA gene is a relevant ARG in the water environ-
ment, including wastewater. The gene was found in nearly
100 % of A. hydrophila and A. jandaei isolates. This confirms
previous observations that many Aeromonas species can nat-
urally produce chromosomal β-lactamases and also suggests
that this resistance could be species-specific (Chen et al.
Ann Microbiol
2012). Moreover, different genes encoding β-lactamases of-
ten associated with mobile genetic elements (MGE) have been
recently detected in Aeromonas species from wastewater, i.e.,
blaTEM, on the plasmid (Figueira et al. 2011; Igbinosa and
Okoh 2012), blaKPC, as a part of transposone Tn4401 (Picão
et al. 2013), and blaPSE1/CARB1, in integron class 1 gene
cassette (Igbinosa and Okoh 2012). These β-lactamases were
found in previous studies in WWTP in various bacterial
genera, confirming at the same time the spread of these genes
through HGT (Li et al. 2009; Pignato et al. 2010; Chagas et al.
2011; Zhang et al. 2012). Most alarming is the occurrence of
carbapenemases genes blaKPC identified in Aeromonas spp.
inhabiting hospital effluent wastewater (Picão et al. 2013).
This observation definitely requires following-up with more
detailed studies.
Moreover, Moura et al. (2007) detected several aminogly-
coside resistance genes in Aeromonas spp. that were isolated
from a slaughterhouse WWTP. All ARG were a part of gene
cassettes, found in class 1 and class 2 integrons. Researchers
have identified aadA1, aadA2 and sat1 genes, encoding the
resistance to streptomycin, spectinomycin and streptothricin,
respectively. The aadA1 and aadA2 genes were also detected
earlier in wastewater bacteria on mobile genetic elements
(MGE) (Szczepanowski et al. 2004; Tennstedt et al. 2005).
Additionally, the trimethoprim resistance gene (dfrA1) has
been also identified in a number of gene cassettes. This
ARG commonly occurs in integron gene cassettes and has
recently been found in Vibrio strains isolated from urban
surface water, and Enterobacteriaceae species from urban
wastewater (Taviani et al. 2008; Pellegrini et al. 2010).
Literature reports show that among tetracycline resistance
genes identified in wastewater, only tetA and tetE have been
found in Aeromonas spp. thus far. It should be emphasized that
both genes code for an efflux pump that removes the drug from
the cell (Chopra and Roberts 2001) and are located on plasmids
(Szczepanowski et al. 2009). Han et al. (2012c) found the tetA
gene in two isolates of A. veronii in South Korea. The study of
Kim et al. (2011) identified the tetE gene in A. salmonicida
isolated from sewage also in Korea. Igbinosa and Okoh (2012)
searched for the tetC gene in their isolates but without success.
Even then, the results of phenotypic studies demonstrated a
higher level of resistance to tetracycline, compared to those
obtained in the molecular studies, by more than 70 % in Fort
Beaufort WWTP, and more than 15 % in Alice WWTP. As a
result, further studies should be carried out to search for other
tet resistance genes commonly found in wastewater (Auerbach
et al. 2007; Zhang et al. 2009b; Marti et al. 2013).
Natural water
A variety of intrinsic and acquired ARG have been identified
in the microorganisms living in natural water bodies, such as
rivers, lakes, estuaries and groundwater (Zhang et al. 2009a).
It is hardly possible to find an aquatic environment without
ARG. Literature reports showed that ARG were detected even
in the pristine sources of rivers, deep groundwater and the
ocean (Batt et al. 2006; Storteboom et al. 2010; Chen et al.
2013). According to recent studies on ancient DNA from
30,000-year-old permafrost sediments, ARG have existed in
the natural environment as equally long as antibiotic pro-
ducers (D'Costa et al. 2012). However, the anthropogenic
impact, in the form of large quantities of exogenous antibi-
otics, has increased the dissemination of ARG in the natural
water environment (Chen et al. 2013; Khan et al. 2013).
The diverse ARG belonging to different groups that are
ubiquitous in various natural waters, were also found in
Aeromonas spp. (Henriques et al. 2006; Picão et al. 2008;
Girlich et al. 2011). Quinolone resistance genes were observed
in lakes and rivers where mutations were found in the quino-
lone resistance-determining regions (QRDR) and plasmid-
mediated transferable quinolone resistance (PMQR) (Cattoir
et al. 2008; Alcaide et al. 2010). Mutations in the gyrA-
encoding subunit were identified with greater frequency than
in the parC gene, suggesting that topoisomerase IV is a
secondary target for quinolones in Aeromonas spp. (Goñi-
Urriza et al. 2002; Han et al. 2012a). The most frequent point
mutation in the gyrA gene is at codon 83, while in the parC
gene it is at codons 80 and 84, which is common among other
gram-negative bacteria (Alcaide et al. 2010). Two types of
PMQR genes have been identified, i.e., qnrS2, encoding
proteins that protect DNA gyrase from quinolones, and
aac(6’)-ib-cr (accA4-cr) (Cattoir et al. 2008; Picão et al.
2008; Majumdar et al. 2011), encoding aminoglycoside ace-
tyltransferase with the amino acid substitutions (Trp102Arg
and Asp179Tyr) necessary for the ability to acetylate cipro-
floxacin (Park et al. 2006). Both genes were previously de-
tected in Aeromonas spp. in wastewater (discussed above),
indicating that quinolone resistance genes are widespread in
the water environment. This could be explained by the use of
quinolones as the drug of first choice in the treatment of
Aeromonas infections in clinical practice (Goñi-Urriza et al.
2000).
The most numerous group of ARG among Aeromonas
species in natural water are β-lactam ARG (Table 1). This is
associated with the increasing resistance to β-lactam antibi-
otics among this genus, which is also observed in clinical
isolates (Aravena-Román et al. 2012). A chromosomally
encoded β-lactamase cphA has been found in A. hydrophila
isolate from estuarine water, as has already been observed in
Aeromonas strains from wastewater (Henriques et al. 2006).
Moreover, recent studies have also shown the occurrence of
plasmid-carried blaFOX genes, enconding AmpC enzyme, in
natural water (Maravić et al. 2013; Voolaid et al. 2013).
Maravić et al. (2013) studied A. caviae strains isolated from
a marine mussel and found only a few plasmid-encoded
 Ann Microbiol

Table 1 Antibiotic resistance genes in different types of environment

Resistance Resistance Biological source Enviromental source* Reference

gene phenotype

accA4-cr quinolones A. media, A. allosaccharophila WWTP (TWW, RWW), Picão et al. 2008; Figueira et al. 2011
WTP, NW (L)
qnrS2 quinolones A. media, A. allosaccharophila, WWTP (TWW, RWW), Cattoir et al. 2008; Picão et al. 2008;
A. caviae, A. veronii, A. hydrophila, WTP, NW (L, R, F), AQ Verner-Jeffreys et al. 2009; Ishida
A. sobria et al. 2010; Figueira et al. 2011;
Majumdar et al. 2011; Han et al. 2012b
gyrA quinolones A. allosaccharophila, A. aquariorum, WWTP (TWW, RWW), WTP, Goñi-Urriza et al. 2002; Alcaide et al. 2010;
(mutation) A. hydrophila, A. media, A. caviae, NW (L, R), F Figueira et al. 2011; Kim et al. 2011;
A. sanarellii, A. taiwanesis, Han et al. 2012a
A. veronii, A. sobria,
A. salmonicida,
A. popoffii
parC quinolones A. allosaccharophila, A. aquariorum, WWTP (TWW, RWW), Goñi-Urriza et al. 2002; Alcaide
(mutation) A. hydrophila, A. media, A. veronii, WTP, NW (R, F) et al. 2010; Figueira et al. 2011
A. caviae, A. taiwanesis,
A. salmonicida
aadA1 aminoglycosides Aeromonas sp., A. caviae, A. media WWTP (TWW, EW), Henriques et al. 2006; Moura et al. 2007;
A. hydrophila NW (M, F), HC, AQ Verner-Jeffreys et al. 2009; Ishida
et al. 2010; Carvalho et al. 2012
aadA2 aminoglycosides Aeromonas sp., A. caviae, WWTP (TWW), NW Henriques et al. 2006; Moura et al. 2007;
A. hydrophila, A. eucrenophila, (M, R, F), HC, AQ (S, F) Verner-Jeffreys et al. 2009; Ndi and
A. media, A. veronii, A. salmonicida Barton 2011; Carvalho et al. 2012
aadA7 aminoglycosides A. salmonicida AQ McIntosh et al. 2008
sat2 aminoglycosides Aeromonas sp. WWTP (EW) Moura et al. 2007
aadA1a aminoglycosides A. hydrophila, A. salmonicida, AQ (F,S) Schmidt et al. 2001; Jacobs and
A. sobria, A. veronii, A. isthiosomia, Chenia 2007
A. encheleia, A. bestiarum
accA4 aminoglycosides A. caviae, A. hydrophila AQ (F) Verner-Jeffreys et al. 2009
strA-strB aminoglycosides A. salmonicida, A. caviae, AQ, NW (R, S, F) Gordon et al. 2008; McIntosh et al.
A. bestiarum 2008; Verner-Jeffreys et al. 2009
aacA aminoglycosides A. hydrophila, A. salmonicida, AQ Jacobs and Chenia 2007
A. isthiosomia, A. sobria,
A. encheleia
cphA/imiS β-lactams A. hydrophila, A. veronii, A. jandei, WWTP (TWW), WTP, Figueira et al. 2011;
A. allosaccharophila, A. aquariorum, NW (M), HC Henriques et al. 2006; Balsalobre
A. media, A. bestiarum, et al. 2009; Carvalho et al. 2012
A. eucrenophila, A. salmonicida
blaKPC-2 β-lactams Aeromonas spp. WWTP, SW Picão et al. 2013
blaVEB-1a β-lactams A. caviae, A. allosaccharophila, NW (R) Girlich et al. 2011
A. veronii, A. media
blaSHV-12 β-lactams A. allosaccharophila, A. veronii, NW (R, M) Girlich et al. 2011; Maravić et al. 2013
A. media, A. caviae, A. hydrophila
blaPER (1,6) β-lactams A. allosaccharophila, NW (R, M) Girlich et al. 2010; Girlich et al. 2011;
A. veronii, A. media, A. caviae Maravić et al. 2013
blaGES-7 β-lactams A. veronii NW (R) Girlich et al. 2011
blaTLA-2 β-lactams A. allosaccharophila NW (R) Girlich et al. 2011
blaOXA (2,7) β-lactams A. hydrophila, A. sobria, NW (M, F), AQ Henriques et al. 2006;
A. salmonicida, A. caviae, Jacobs and Chenia 2007;
Verner-Jeffreys et al. 2009
blaTEM β-lactams Aeromonas sp., A. media, A. veronii, WWTP (RWW), Henriques et al. 2006; Pontes et al. 2009;
A. hydrophila, A. caviae NW (M, R, L, F), HC Verner-Jeffreys et al. 2009; Figueira
et al. 2011; Carvalho et al. 2012;
Igbinosa and Okoh 2012; Tacão et al. 2012
blaCTX-M β-lactams A. hydrophila, A. caviae NW (R, M) Tacão et al. 2012; Maravić et al. 2013
blaPSE-1/CARB-1 β-lactams Aeromonas sp., A. isthiosomia, WWTP, AQ Jacobs and Chenia 2007; Igbinosa
A. sobria, A. encheleia, A. hydrophila and Okoh 2012
blaCMY-2 β-lactams A. salmonicida AQ McIntosh et al. 2008
blaFOX β-lactams Aeromonas sp., A. caviae NW (R, M) Maravić et al. 2013; Voolaid et al. 2013
Ann Microbiol

Table 1 (continued)

Resistance Resistance Biological source Enviromental source* Reference

gene phenotype

dfrA1/7 trimethoprim Aeromonas sp., A. media, A. sobria, WWTP (TWW, EW), Jacobs and Chenia 2007; Moura et al. 2007;
A. salmonicida, A. hydrophila HC, AQ, NW (F) Verner-Jeffreys et al. 2009; Ishida
et al. 2010; Carvalho et al. 2012
dfrA12 trimethoprim A. hydrophila, NW (estuarine, F), Henriques et al. 2006; Ishida et al. 2010;
A. media, A. veronii HC, AQ Carvalho et al. 2012; Verner-Jeffreys
A. eucrenophila, A. caviae et al. 2009
dfr13 trimethoprim A. hydrophila AQ (F) Verner-Jeffreys et al. 2009
dhfr2a trimethoprim A. hydrophila AQ, S Schmidt et al. 2001
dhfr1 trimethoprim A. hydrophila, A. sobria, A. veronii F, AQ, S Schmidt et al. 2001
A. bestiarum
sul1 sulphonamides A. salmonicida, A. veronii, A. caviae, AQ (S, F) McIntosh et al. 2008; Verner-Jeffreys
A. hydrophila et al. 2009; Ndi and Barton 2011
sul2 sulphonamides A. salmonicida, A. bestiarum AQ, NW (R,S) Gordon et al. 2008; McIntosh et al. 2008
estX streptothricin Aeromonas sp. WWTP (EW) Moura et al. 2007
cat chloramphenicol Aeromonas sp. NW (M) Dang et al. 2008
catB2 chloramphenicol A. hydrophila, A. sobria, A. veronii, F, AQ (S) Schmidt et al. 2001
A. bestiarum
catB3 chloramphenicol A. allasaccharophila, A. hydrophila NW (L) AQ Picão et al. 2008; Ishida et al. 2010
catB8 chloramphenicol A. hydrophila, A. caviae NW (M), HC, AQ (F) Henriques et al. 2006; Verner-Jeffreys
et al. 2009; Carvalho et al. 2012
floR chloramphenicol A. salmonicida, A. hydrophila, AQ (F), NW (R, S) Gordon et al. 2008; McIntosh et al. 2008;
A. caviae, A. bestiarum Verner-Jeffreys et al. 2009
tetA tetracycline A. hydrophila, A. allosaccharophila, WWTP, HC, AQ (F), Schmidt et al. 2001; Nawaz et al. 2006;
A. veronii, A. sobria, A. salmonicida, F, S, NW (R) Akinbowale et al. 2007; Jacobs and
A. encheleia, A. media, A. icthiosomia, Chenia 2007; McIntosh et al. 2008;
A. caviae Verner-Jeffreys et al. 2009; Girlich
et al. 2010; Ishida et al. 2010;
Girlich et al. 2011; Kim et al. 2011;
Carvalho et al. 2012; Han et al. 2012c
tetB tetracycline A. hydrophila, AQ Nawaz et al. 2006; Jacobs and
A. encheleia, A. veronii Chenia 2007
tetC tetracycline A. hydrophila, HC, AQ, S, F Nawaz et al. 2006; Jacobs and
A. veronii, A. caviae Chenia 2007; Verner-Jeffreys et al.
2009; Ishida et al. 2010; Ndi and
Barton 2011; Carvalho et al. 2012;
Han et al. 2012c
tetD tetracycline Aeromonas sp., A. hydrophila, HC, AQ, S, Schmidt et al. 2001; Nawaz et al. 2006;
A. encheleia, A. veronii, A. sobria F (AQ) Akinbowale et al. 2007; Jacobs and
A. salmonicida, A. caviae Chenia 2007; Verner-Jeffreys et al. 2009;
Carvalho et al. 2012; Han et al. 2012c
tetE tetracycline A. eucrenophila, A. hydrophila, WWTP, HC, AQ, S, F, Schmidt et al. 2001; Nawaz et al. 2006;
A. icthiosomia, A. sobria, A. encheleia, F (AQ) Akinbowale et al. 2007; Jacobs and
A. veronii, A. caviae, A. salmonicida Chenia 2007; Verner-Jeffreys
et al. 2009; Ishida et al. 2010;
Kim et al. 2011; Carvalho et al. 2012;
Han et al. 2012c
tetG tetracycline A. caviae AQ (F) Verner-Jeffreys et al. 2009
tetH tetracycline A. hydrophila, A. encheleia AQ Jacobs and Chenia 2007
tetM tetracycline A. hydrophila, A. sobria, AQ (F) Akinbowale et al. 2007
A. hydrophila
tetY tetracycline A. bestiarum NW Gordon et al. 2008
vatE streptogramin A. caviae AQ (F) Verner-Jeffreys et al. 2009

Footnotes: *WWTP- wastewater treatment plant; RWW- raw wastewater; TWW- treated wastewater; WTP- water treatment plant; EW – effluent water;
SW- special wastewater from hospital; NW- natural water; HC – human consumption; AQ- aquacultures; F- fishes; S- sediments; L- lakes, R-rivers; M-
marine waters, estuaries

blaFOX-2 genes. However, the study of Voolaid et al. (2013)
suggested that the dissemination of blaFOX genes might be
significantly more widespread in environmental bacteria, with
a great contribution of Aeromonas. Previous studies demon-
strated that Aeromonas were natural producers of chromosom-
al AmpC and MBL β-lactamases (Chen et al. 2012).
It is suspected that some of these chromosomal AmpC
enzymes are ancestors of FOX β-lactamases (Janda and
Abott 2010). In addition to those listed above, numerous
Extended-Spectrum β-lactamases (ESBL) such as blaVEB,
blaSHV, blaPER, blaGES, blaTLA, blaOXA, blaTEM and blaCTX-M
(Henriques et al. 2006; Girlich et al. 2010; Girlich et al. 2011;
Maravić et al. 2013) have been found in Aeromonas species.
ESBL do not belong to intrinsic determinants of Aeromonas
spp. but bacteria acquire them from the environment where
other ARB, e.g., representatives of the Enterobacteriaceae,
among which ESBL-mediated resistance is widespread, are
ubiquitous (Blaak et al. 2013). Depending on the gene, the
location might be different, but generally bla genes were iden-
tified within MGE even when they were found in chromosomes.
For example, blaSHV-12, blaPER-1 and blaPER-6 were localized
nearby insertion sequences and transposons (Girlich et al. 2011).
The blaVEB-1a genes were identified on plasmids and chromo-
somes, even though certain ESBL were solely attributed to
plasmids, e.g., blaGES-7 and blaTLA-2. The majority of these
genes such as blaOXA, are also part of the class 1 integron
cassettes (Henriques et al. 2006). According to recent studies,
the most widespread ESBL genes among Aeromonas spp. are
blaTEM and blaCTX-M, which are detected in lakes, rivers and
estuarine water (Henriques et al. 2006; Pontes et al. 2009; Lu
et al. 2010; Tacão et al. 2012). Both of them are plasmid-
mediated, and in the study of Tacão et al. (2012) they were
found more frequently in polluted sites. The researchers suggest
that these findings sustain the hypothesis concerning anthropo-
genic modulation of resistance in environmental bacteria.
Aminoglycoside resistance genes from natural waters have
been identified thus far in Aeromonas spp. in two studies.
According to the literature, aadA1, aadA2 and strA-strB were
detected in A. caviae, A. hydrophila and A. bestiarum in
estuarine and river environments (Henriques et al. 2006;
Gordon et al. 2008). All these ARG were found on MGE-
like plasmids and very often were a part of integron cassettes.
Sulfonamides and trimethoprim resistance genes, sul and dfr,
are a commonly occurring type of ARG in natural water (Stoll
et al. 2012; Suzuki et al. 2013). Similarly, as in the case of
aminoglycoside resistance, a couple of ARG have been iden-
tified in Aeromonas spp. only in the studies of Henriques et al.
(2006) and Gordon et al. (2008). The only identified genes
were sul2 and dfrA12. Trimethoprim resistance genes (dfr),
encoding dihydrofolate reductases and sulfonamide ARG
sul1, were frequently found as part of integron cassettes. The
sul2 gene has so far been found only once in A. bestiarum as a
plasmid-mediated gene (Gordon et al. 2008).
Ann Microbiol
According to the current literature, the resistance to chlor-
amphenicol and florfenicol among Aeromonas spp. has been
identified quite often. The cat genes (cat, catB3 and catB8),
encoding chloramphenicol resistance, were found in seawa-
ters and lakes (Henriques et al. 2006; Dang et al. 2008; Picão
et al. 2008). Florfenicol/chloramphenicol resistance gene (floR)
was found in isolates from river sediments (Gordon et al. 2008).
Moreover, the resistance to chloramphenicol was observed
among various environmental bacteria (particularly in
Pseudomonas), especially in marine water (Dang et al. 2008).
In the environment, both groups of the tet genes have been
identified in a number of bacterial species. They code for
ribosomal protection proteins and efflux pumps (Zhang et al.
2009b), often associated with conjugative and mobilizable
elements, such as plasmids, transposons and integrons
(Chopra and Roberts 2001; Schmidt et al. 2001). Aeromonas
spp. isolated from natural waters frequently demonstrated a
tetracycline resistance phenotype (Goñi-Urriza et al. 2000;
Tacão et al. 2012). However, until now only two molecular
determinants of this resistance have been found, i.e., the tetA,
and tetY genes that encode efflux pump proteins and are
located on the transposon and plasmid, respectively (Gordon
et al. 2008; Girlich et al. 2011). Additionally, the tetracycline
resistance genes occur together with the tetR gene that en-
codes the tetracycline repressor protein, located on Tn10
(Meier et al. 1988). The study of Gordon et al. (2008) revealed
for the first time an association of the tetY and tetR genes,
which was identified on a plasmid of A. bestiarum isolated
from freshwater sediments in France. More recently, the tetA-
tetR genes were identified by Girlich et al. (2010, 2011) who
found them in A. allosaccharophila and A. veronii, isolated
from another river in France. The prevalence of these genes
should definitely be investigated in the future, because
Aeromonas can be a unique vector spreading them in the water
environment.
Aquacultures
Farming of aquatic organisms (aquacultures) creates another
specific water environment, due to the great anthropogenic
impact. Large amounts of antibiotics are added into the water
with feed as prophylactic and therapeutic agents. The most
widely used drugs are fluoroquinolones, florfenicol, oxytetra-
cyclines (OTC), amoxicillin and sulfonamides (Gräslund et al.
2003; Holmström et al. 2003; Cabello 2006; Primavera 2006;
Soonthornchaikul and Garelick 2009). Consequently, resis-
tance to antibiotics has been widely spread in aquaculture
environments, which has been reported in many studies
(Petersen et al. 2002; Colquhoun et al. 2007; Newaj-Fyzul
et al. 2008; Sørum 2008; Shah et al. 2012). As a result of these
concerns, the usage of antibiotics as growth promoters has
been strictly regulated or even banned in many countries, e.g.,
Ann Microbiol
in European Union from January 1, 2006 (Regulation (Ec) No
1831/2003). Nevertheless, uncontrolled and continuous long-
term use of antibiotics for therapeutic and prophylactic rea-
sons also had a major influence on increased transfer of ARG
between microorganisms (Tennstedt et al. 2003;
Szczepanowski et al. 2009; Cheng et al. 2012; Moura et al.
2012). Among ARB in aquacultures are also fish pathogens
and human opportunistic pathogens, thus, it poses a particu-
larly serious threat to human health. Resistant bacteria can be
transferred to humans with infected fish or by direct contact
with aquaculture ecosystems such as in the case of fish farm
workers (Petersen and Dalsgaard 2003). Antimicrobial resis-
tance has long been reported in widespread Aeromonas spe-
cies in aquacultures, and recently an upward trend in the
resistance has been observed (Rhodes et al. 2000; Schmidt
et al. 2001; Jacobs and Chenia 2007). The study of Navarrete
et al. 2008 showed loss of diversity for the microbiota of fish
treated with OTC, due to eradication of competing microor-
ganisms, and included only Aeromonas spp. Disturbing epi-
demiological and molecular data indicate that Aeromonas
strains are able to transmit and share AMR determinants with
such bacteria as Escherichia coli isolated from humans
(Rhodes et al. 2000; Sørum 2006).
The largest group of ARG identified in aquaculture isolates
of Aeromonas species is the tet genes. Among many tetracy-
cline resistance genes, eight classes of the tet genes (tetA, tetB,
tetC, tetD, tetE, tetH, tetG and tetM) have been found in
Aeromonas spp. isolated from water, sediments and diseased
fish in aquaculture systems (Table 1) (Schmidt et al. 2001;
Nawaz et al. 2006; Akinbowale et al. 2007; Jacobs and Chenia
2007; Verner-Jeffreys et al. 2009). Five of these genes are
located on the plasmid (tetA, tetB tetC, tetD, tetE), one in the
transposon (tetA) and two (tetC and tetE) are adjacent to the
integrons as part of the same transposons. Most of them
encode efflux pump proteins but the gene tetM, encoding a
ribosomal protein has been also found (Akinbowale et al.
2007). The results in the literature indicate that the tetA and
tetE determinants are the predominant tetracycline resistance
genes (Nawaz et al. 2006; Han et al. 2012c). The study by
Schmidt et al. (2001), performed on a collection from a
Danish rainbow trout farm, showed that the tetE gene was
the most frequently identified after the tetA and tetD genes.
What is more, in some instances, more than one tet gene from
different classes was observed in a single isolate. More recent
studies of Nawaz et al. (2006) and Han et al. (2012c), who
isolated Aeromonas spp. from commercial catfish ponds in
Texas and Korean fish farms and aquariums, respectively, also
confirmed the tetE gene as the predominant agent. However,
Kim et al. (2011), who isolated A. salmonicida strains from
salmonid farms and private fish tanks in Korea, reported that
tetA was the most frequent gene. Ndi and Barton (2011) who
conducted their study on rainbow trout farms in Australia
found the tetA genes in some isolates, but tetC was the
predominant agent in all tetracycline resistant Aeromonas
isolates. In the same study, two or even three genes encoding
tetracycline resistance were identified in a single Aeromonas
isolate but it did not correlate with higher MIC. The study of
Akinbowale et al. (2007) also investigated Australian aqua-
cultures and found that the tetM genes were the most preva-
lent, and in all cases were found in combination with more
than one tet gene. These results suggest that the tet genes are
common in aquacultures, but depending on the source of
isolation, various combinations of these genes can be found.
Certain studies also indicated that the tet determinants are
associated with conjugative plasmids (Schmidt et al. 2001).
Resistance to aminoglycosides in Aeromonas spp. isolates
from aquacultures is diverse and additionally depends on the
origin of probes. Dias et al. (2012) who studied samples of
water and skin of ornamental fish obtained from a national
importer, reported the resistance to aminoglycosides in 6-30 %
of isolates depending on the antibiotic. However, Hatha et al.
(2005), in their study on samples from freshwater fish farms,
observed this resistance at a much lower level (0-7 %). Nev-
ertheless, many aminoglycoside resistance genes have been
found thus far in Aeromonas spp. isolated from this environ-
ment. Furthermore, genes encoding aminoglycoside-
modifying enzymes, such as aadA1a, aadA2, aadA7, aacA4,
aacA and streptothricin phosphoryltransferase strA and strB
have also been detected (Jacobs and Chenia 2007; McIntosh
et al. 2008; Verner-Jeffreys et al. 2009; Ndi and Barton 2011).
According to the literature, the aad family of genes was
detected most frequently and was localized within potentially
mobile gene cassettes (Verner-Jeffreys et al. 2009; Ndi and
Barton 2011).
The occurrence of antibiotic resistance genes against dif-
ferent groups of antibiotics among Aeromonas spp. derived
from aquacultures, is generally at the similar level and is less
common. Quinolone resistance determinants were identified
in diseased fish from aquacultures (Ishida et al. 2010). Han
et al. (2012a), observed in their study both QRDR mutations
in the gyrA or parC genes as well as plasmid-encoded qnrS
genes. The same authors identified qnrS2 genes in a different
work (2012b), as part of the mobile insertion cassette on ColE-
like plasmids, which were isolated from A. sobria and
A. hydrophila strains. Verner-Jeffreys et al. (2009) also found
qnrS2 genes, and using PCR and microarray techniques,
detected the occurrence of these genes in association with
Inc A/C plasmids.
Β-lactam resistance within this genus in aquacultures is
mainly limited to the widespread ampicillin and amoxicillin
resistance. To date, only a few β-lactamases determinants
have been found and they include: blaOXA, blaPSE-1/CARB-1
and blaCMY-2 (Jacobs and Chenia 2007; McIntosh et al. 2008).
The study by Jacobs and Chenia (2007) found the blaOXA-2a
and blaPSE-1 genes within the gene cassettes of class 1
integrons in almost 6 % and 30 % of isolates, respectively.

In addition, the blaCMY-2 gene, which encodes β-lactamase
from the AmpC group, was identified on a plasmid isolated
from A. salmonicida (McIntosh et al. 2008).
The determinants of chloramphenicol (catB) and
florfenicol (floR ) resistance identified thus far, encode
acetylotransferase and efflux proteins. To date, several types
of catB have been reported, i.e., catB2, catB3 and catB8
(Schmidt et al. 2001; Verner-Jeffreys et al. 2009; Ishida et al.
2010). McIntosh et al. (2008) found the floR gene within an
antibiotic resistance cassette identified in multidrug-resistant
(MDR) A. salmonicida.
The resistance to trimethoprim and sulphonamide was also
observed in this environment among Aeromonas species. This
led to the identification of trimethoprim and sulphonamide
resistance genes, i.e., dihydrofolate reductase encoding differ-
ent types of dfrA genes such as dfrA1 or dfrA7 (Table 1)
(Jacobs and Chenia 2007; Verner-Jeffreys et al. 2009) as well
as a dihydropteroate synthase, encoding the sul1 and sul2
genes (McIntosh et al. 2008; Ndi and Barton 2011). Schmidt
et al. (2001) showed a strong association between the resis-
tance to sulphaziadine/trimethoprim (S/T) and the occurrence
of class 1 integrons. Moreover, all S/T resistance isolates of
Aeromonas (mostly A. hydrophila) contained an integron with
a sul1 gene and dfr gene cassette insert. The follow-up study
by Verner-Jeffreys et al. (2009) confirmed this observation in
Aeromonas isolates, in which the sul1 genes and various gene
cassettes containing dfr determinants were found. The sul2
gene is less prevalent but was also identified as part of a
plasmid-located sequence containing genes floR, tetA/R and
strA/B from MDR A. salmonicida (McIntosh et al. 2008).
Urban water for human consumption
Aeromonas species have been also detected in sources of
treated urban water for human consumption, such as water
treatment plants, wells, tap water or even drinking mineral
water (Scoaris Dde et al. 2008; Figueira et al. 2011; Kivanc
et al. 2011). Some species, e.g., A. hydrophila, A. caviae and
A. sobria are agents of diarrheal episodes, particularly in
immunocompromised people (Ali et al. 2005; Janda and
Abott 2010). For this reason, the occurrence of Aeromonas
spp. in water for human consumption and the levels of resis-
tance and virulence of these strains should be investigated.
There are only a few studies on the occurrence of antibiotic
resistance genes in Aeromonas species isolated from urban
water, thus, our knowledge regarding the occurrence of ARG
in this environment is poor. Moreover, according to the United
States Environmental Protection Agency, the number of
Aeromonas spp. in treated water is typically below 10 cfu/
ml. Therefore, the isolation of bacteria is problematic
(Aeromonas: United States Environmental Protection Agency
2006). Carvalho et al. (2012) isolated Aeromonas spp. from
Ann Microbiol
the following sources of urban water for human consumption:
fountains, mines, wells and drilled wells. A. media, A.
bestiarum and A. hydrophila were the most frequently isolated
species in that study. As expected, all strains were resistant to
amoxicillin and the majority of them to the first generation
cephalosporin, cefalotin, and the carboxypenicillin and
ticarcillin. Searching for β-lactam resistance genes resulted
in the identification of chromosomal cphA/imiS genes in 21
strains, mostly in A. hydrophila. Despite the existence of many
bla genes and their broad distribution, only in one strain of
A. veronii the blaTEM gene was found. The study by Figueira
et al. (2011) also showed that the cphA genes were prevalent in
76 % of strains from a drinking water treatment plant and were
predominant in A. hydrophila, A.veronii and A. jandei. The
same study identified the determinants of resistance to quino-
lones, i.e., the accA4 and qnrS genes as well as a mutation in
the gyrA and parC genes that was observed in the previously
discussed environments. However, the frequency of gyrA
mutations in the water treatment plant was significantly
lower than in the wastewater treatment plant. In addition, the
study of Emekdas et al. (2006), who investigated the tap water
environment, showed that all Aeromonas isolates were sus-
ceptible to ciprofloxacin.
Carvalho et al. (2012) identified the tet genes in 10 % of
isolates, which corresponded with a low level of tetracycline
resistance. All of the genes observed encoded efflux pump
proteins (tetA, tetC, tetD and tetE) and were located on the
plasmid. The most frequent was the tetE gene and the majority
of them were found in A. hydrophila. The study by Scoaris
Dde et al. (2008) observed a higher level of resistance to
tetracycline (26 %) but did not prove the presence of ARG
using molecular procedures. The same level of resistance was
observed in the case of trimethopim-sulfamethoxazole and
gentamicin. However, Carvalho et al. (2012) found several
aminoglycoside and trimethoprim resistance genes (aadA1,
aadA2, dfrA1 and dfrA12). They were identified in different
configurations in gene cassettes of group I and II integrons.
Additionally, the catB8 gene has been found that encoded
resistance to chloramphenicol. Surprisingly, all of these po-
tentially mobile genes were found in A. hydrophila.
Resistance genes among Aeromonas spp. localized
on mobile genetic elements
MGE such as plasmids and transposons are vectors carrying
resistance genes, which are widespread in many different
environments. They allow the exchange of antibiotic and
metal resistance genes among microorganisms through
conjugative transfer, transformation or transduction. As a
result, they play a significant role in the HGT between various
bacterial species, including those phylogenetically distant
(Martinez et al. 2009). Of most concern, is the dissemination
Ann Microbiol
of the broad-host range (BHR) and conjugative plasmids.
These MGE can transfer ARG from environmental species
to clinically relevant pathogens (Rhodes et al. 2000; Sørum
2006). A water environment is a significant reservoir of mi-
croorganisms that possess mobile resistant genes. MGE car-
rying resistance genes are also widespread among Aeromonas
spp. (Table 2). BHR are the most frequently identified plas-
mids in Aeromonas spp., and they belong to two different
incompatibility groups, i.e., Inc A/C and IncU. The majority
of them are also MDR and are capable of self-conjugative
transfer (tra genes) or a transfer by mobilization (mob genes).
Del Castillo et al. (2013) recently identified a 165 kb pR148
plasmid from A. hydrophila. This large plasmid belongs to the
IncA/C group, distinguished by MDR, conserved backbone
and association with enteric human pathogens (Johnson and
Lang 2012). The plasmid identified consisted of resistance
genes to seven different pollutants within two transposon
types, Tn21 and Tn1721. In addition to antibiotic resistance
to β-lactams, sulfonamides, tetracycline, chloramphenicol
and aminoglycosides, pR148 carried genes encoding resis-
tance to mercury and quaternary ammonium compounds.
More importantly, phylogenetic comparative studies revealed
that pR148 is most closely related to human pathogenic E. coli
and Acinetobacter baumanii. This similarity indicates that the
IncA/C group of plasmids was transferred between different
genera. Another study also identified several plasmids from
this group carrying MDR and capable of conjugative transfer
(McIntosh et al. 2008; Moura et al. 2012).
The second type of plasmids that belongs to the IncU group
is probably even more widespread in the environment. These
BHR R-plasmids also have a highly conserved backbone and
carry various antibiotic gene cassettes. For instance, closely
related plasmids pRAS1, pFBOAT4 and pASOT, which were
isolated from different Aeromonas species, all contained tetA
genes and were capable of conjugative transfer between
humans and aquaculture environments (Rhodes et al. 2000).
Moreover, MDR plasmids such as pRAS1 and pPAR-32 that
share the same core genes but have different integron cassettes
(In4-like and In6-like), carrying various resistance genes, also
belong to the IncU group (Sørum et al. 2003). Furthermore,
the IncU plasmids are also associated with the qnrS genes,
which have been found in different Aeromonas species all
around the world (Cattoir et al. 2008; Picão et al. 2008).
Transposons (Tn) and insertion sequences (IS) carrying
ARG have been described in many studies (Table 2). The
fragments of Tn1721 were identified also in chromosome-
like plasmids that carried the tetA and tetR genes (Sørum
et al. 2003; Girlich et al. 2011). This transposon has been
previously found in various bacteria also belonging to human
pathogens, e.g., Salmonella enterica and hospital isolates of
E. coli (Popowska and Krawczyk-Balska 2013). What is also
worrying from the point of view of public health is the
prevalence of blaKPC resistance genes harbored on another
type of transposon, Tn4401. These MGE have been identified
within conjugative plasmids isolated from Aeromonas spp.
inhabiting hospital sewage (Picão et al. 2013). Additionally,
many ARG among Aeromonas spp., mostly encoding β-
lactamases, were identified within various IS, e.g., IS26,
ISpa12, ISpa13, IS6100 or ISEcp1 (Table 2). Moreover, two
insertion sequence common region elements (ISCR), called
ISCR2, were found on the conjugative plasmid pAB5S9
(Gordon et al. 2008). These MGE are closely related to the
family of IS91 insertion sequences and are considered to play
an important role in the acquisition of ARG (Toleman et al.
2006). However, participation of ISCR in gene transfer among
Aeromonas spp. at this time seems to be secondary and should
be fully explored in the future.
Bacteriophages also have to be taken into consideration in
the HGT of ARG among Aeromonas spp. Until now, over a
dozen of Aeromonas spp. phages have been isolated from
different environments, e.g., A. hydrophila phages Aeh1 and
Aeh2 from sewage (Chow and Rouf 1983), A. hydrophila
phage CC2 from sewage in China (Shen et al. 2012),
A. salmonicida phage phiAS4 from a river in Korea (Kim
et al. 2012a) and A. salmonicida phage PAS-1 from aquacul-
ture in Korea (Kim et al. 2012b). Prophages have also been
found that can contribute to HGT following excision and
transduction. Five putative prophages associated with viru-
lence genes were detected in A. hydrophila isolated from an
epidemic outbreak of catfish in the USA (Hossain et al. 2013).
One of them, the prophage AH2, shared a significant homol-
ogy with ΦO18P and ΦO18P-like prophages of A. caviae
(Ae398) and A. salmonicida (A449) (Hossain et al. 2013).
Beilstein and Dreiseikelmann 2008 described the prophage
ΦO18P (Myoviridae) in A. media isolated from a pond in
Germany. Despite these reports, the participation of bacterio-
phages in the development of antibiotic resistance has never
been demonstrated in Aeromonas spp. thus far.
Finally, resistance genes in Aeromonas spp. are often adja-
cent to integrons or even exist within integron cassettes, and
they can be located either on chromosomes or MGE. Many
different ARG cassettes have been found among Aeromonas
spp. in various water environments, including integron classes
1 and 2 (Table 2). Class 1 from integrons was identified with
higher frequency.
Conclusions
Resistance to various pollutants, and antibiotics in particular,
is broadly distributed among environmental bacteria, includ-
ing Aeromonas spp. In this review, we focused on the preva-
lence of ARG in natural water ecosystems as well as in those
with anthropogenic influence. Recent literature has reported
the identification of genes encoding resistance to many differ-
ent groups of antibiotics in aquatic environments that include
 Ann Microbiol

Table 2 Aeromonas spp. resistance genes on motile genetic elements and integrons find in literature data

Motile genetic Antibiotic resistance genes Enviromental Others References

elements source **

pR148 MDR*: − Tn21: qacH, blaOXA-10, AQ (F) 165 kb; IncA/C; intI Castillo et al. 2013
aadA1, sul1
- Tn1721: tetA, tetR
- catA2, sul1, mer
p34 -IS: qnrS2, NW (L) 80 kb; IncU Picão et al. 2008
- intI: aac(6’)-Ib-cr,
blaOXA-1, catB3
pAS37, P42 qnrS2 NW (R) 55/20 kb; IncU Cattoir et al. 2008
pTf28 blaGES-7 NW (R) 60-kb; intI; IR from Tn402; mobA Girlich et al. 2011
R-plasmids Tet, dhfr, catB, aadA1a AQ Large plasmids (>30 kb), different Schmidt et al. 2001
resistance profiles, conjugative
tranfser; intI
pAB5S9 MDR: floR, sul2, NW (R, S) 24,7 kb; ISCR2; conjugative transfer Gordon et al. 2008
strA-strB, tetR-tet(Y)
pAG2 qnrS2 AQ 6,9 kb; type ColE; mobA; mobile IS Han et al. 2012b
pBRST7.6 qnrS2 F 7,6 kb; IncQ; mobC, mobB Majumdar et al. 2011
Plasmid IncFIB blaCTX-M-15 NW 40 kb Maravić et al. 2013
Plasmid IncA/C aadA7, floR, sul 2, NW ~ 11 kb; intI McIntosh et al. 2008
tetA, strA-B, blaCMY-2
pRAS1 MDR: dfrA16,sulI, tetA F 45-kb; IncU; conjugative transfer; Sørum et al. 2003
Tn1721; intI; In4-like integron, IS6100
pRAS3 tetC AQ (F) 11 kb; non-conjugative L’Abée-Lund et al. 2002
pAR-32 MDR: aadA2, sul1, F incU; intI; In6-like integron Sørum et al. 2003
catAII
pAHH01 tetE, tetR(E) NW (R,L,F), 8,9 kb; mob genes; ColE-like replication Han et al. 2012c
WWTP system; relE/B (toxin-antitoxin)
pFBOAT1-17 Tn1721: tetA, tetR(A) AQ, SW ~85 kb; IncU; conjugative transfer; intI, Rhodes et al. 2000;
(pFBOAT4) IS61100, Tn1721-like region, In4-like Rhodes et al. 2004
pASOT tet A-E genes AQ Conjugative transfer; (Tn1721 – 5.4 kb Adams et al. 1998
EcoRI fragment)
Tn1721 blaPER-6 NW (R) Chromosomal location, previously Sørum et al. 2003;
(fragments) reported Girlich et al. 2011
Tn4401 blaKPC-2 WWTP (SW) Putative plasmid location Picão et al. 2013
(conjugative or mobilizable)
IS26 blaSHV-12 NW (R) Previously reported Girlich et al. 2011
blaVEB-1
ISpa12, ISpa13 blaPER-1 NW (R) Chromosomal location Girlich et al. 2011
(Tn1213)
ISCR2, IS6100 blaVEB-1 NW (R) Plasmid location Gordon et al. 2008;
Girlich et al. 2011
ISEcp1 bleCTX-M NW (R) - Tacão et al. 2012
Integrons
integron class 1 aadA2; dfrA1-aadA1; aadA1; WWTP, NW Putative location: chromosomal, plasmid; Schmidt et al. 2001; Henriques
dfrA12-aadA2; catB8-aadA1; (M), HC, et al. 2006; Jacobs and Chenia 2007;
aadA1-blaOXA-2; dfrA12; NW (R), Moura et al. 2007; Verner-Jeffreys
blaTLA-2; blaVEB-1; dfrA5- dfrA1- AQ et al. 2009; Ishida et al. 2010; Girlich
dfrA27-qacE2; aadA1-aadA2; et al. 2011; Ndi and Barton 2011;
dfrA22- dfrA1; dfrA21-dfrA22; Carvalho et al. 2012; Igbinosa and
dfrA23; tetC/E/catB3; qnrS; Okoh 2012; Tacão et al. 2012; Maravić
blaTEM1; dfrA7; tetC; et al. 2013
catB3-aadA1
integron class 2 estX-sat2-aadA1; WWTP, HC, Putative location: chromosomal Jacobs and Chenia 2007;
dfrA1-aadA1; AQ, NW Moura et al. 2007;
Carvalho et al. 2012;
Maravić et al. 2013

Footnotes: * MDR-multidrug resistance; **WWTP- wastewater treatment plant; RWW- raw wastewater; TWW- treated wastewater; WTP- water
treatment plant; EW – effluent water; SW- special wastewater from hospital; NW- natural water; HC – human consumption; AQ- aquacultures; F- fishes;
S- sediments; L- lakes, R-rivers; M-marine waters, estuaries
Ann Microbiol
quinolones, aminoglycosides, β-lactams, tetracyclines, sul-
fonamides, trimethoprim and chloramphenicol.
There are differences in the contribution of resistance to
antibiotics among Aeromonas spp. derived from particular
environments. As a result, certain ARG have been identified
with a higher frequency than others. The most diverse envi-
ronments in terms of antibiotic resistance are natural water and
aquacultures. However, an aquaculture was the only environ-
ment where no Aeromonas strains were detected that would
carry mutations in the gyrA and parC genes, responsible for
the resistance to quinolones. In addition, these strains were
also lacking the chromosomal gene cphA, encoding the
metallo-beta-lactamase. Therefore, it seems that the three
aquatic environments, i.e., WWTP, natural and urban waters,
possess common features such as a low level of antibiotic
pressure on Aeromonas spp. In contrast, aquacultures utilize
therapeutically or preventively high concentrations of antibi-
otics (particularly OTC), which favors the process of HTG
rather than the introduction of chromosomal mutations.
The most numerous group of ARG in natural water are β-
lactamase genes, especially those belonging to the type ESBL.
The most dominant in aquacultures are tetracycline resistance
genes, which are generally widespread in this environment.
There is not enough data in the case of wastewater and
drinking water to draw generalized conclusions, even though
recent results indicate that the occurrence of ARG among
Aeromonas spp. in these environments is also high.
The presence of ARG among A. hydrophila, A. caviae and
A. sobria species, which are considered opportunistic pathogens
responsible for human infections, may represent a major concern
for human health. Moreover, data collected indicate that
Aeromonas spp. are able to share and transfer ARG between
different genera of bacteria, including those isolated from
humans, e.g., E. coli (Cantas et al. 2013). The mechanism of
ARG transmission is not fully understood, but the results of the
studies have suggested that the conjugative broad host range
(BHR) plasmids from IncA/C and IncU have a major contribu-
tion to this process. The co-existence of metal resistance genes
(MRG) and ARG on BHR plasmids have been observed in
numerous previous studies on different bacteria (Popowska and
Krawczyk-Balska 2013), including Aeromonas spp. Literature
data indicate that integrons also greatly contribute to the spread
of ARG among bacteria, including Aeromonas spp., with class 1
being the most prevalent. These small non-replicative genetic
elements cannot transfer ARG by themselves but are spread
among bacteria by MGE. Integron resistance cassettes have
been found in all investigated water environments and the most
prevalent ARG were aadA and dfrA, encoding aminoglycoside
and trimethoprim resistance genes, respectively. Overall, the
analysis of literature data concerning the location of ARG in
Aeromonas strains isolated from various aquatic environments
indicated that plasmids and transposons are more likely to
contribute to ARG dissemination in Aeromonas species.
Infections caused by Aeromonas spp. do not directly pose a
significant threat to public health. However, the transfer of
ARG causes ineffectiveness of antibiotic treatment of fish
diseases, and leads to significant economic losses, particularly
in aquacultures. In conclusion, given the role of Aeromonas
spp. as ARG vectors between environmental bacteria and
clinical human pathogens, as well as the existence of MDR
Aeromonas strains, and the location of ARG on MGE, the
presence of resistant Aeromonas strains in the environment
poses a serious threat to human health.
Acknowledgments The authors’ research was financed by a grant from
the National Center of Science, Poland (741/N-COST/2010/0). The re-
search was conducted as part of the European Co-operation in the field of
Scientific and Technical (COST) Research Action TD0803 “Detecting
evolutionary hot spots of antibiotic resistances in Europe (DARE)”
(2009–2013).
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