Distribution of Antimicrobial Resistance Genes and Class 1 Integron Gene Cassette Arrays in Motile Aeromonas spp. Isolated from Goldfish ( Carassius auratus )
Restricted accessResearch articleFirst published online October, 2018
Distribution of Antimicrobial Resistance Genes and Class 1 Integron Gene Cassette Arrays in Motile Aeromonas spp. Isolated from Goldfish ( Carassius auratus )
Aeromonas spp. are opportunistic pathogens related to multiple infectious diseases in ornamental fishes. In the present study, the antimicrobial susceptibility, resistance genes, and integrons of 65 goldfish-borne Aeromonas spp. were evaluated. The isolates were identified as A. hydrophila (n = 30), A. veronii (n = 32), and A. punctata (n = 3) by gyrB sequencing. The antimicrobial susceptibility testing of the isolates designated that most of the isolates were resistant to amoxicillin (100.00%), nalidixic acid (100.00%), ampicillin (98.46%), tetracycline (92.31%), rifampicin (86.15%), and cephalothin (61.54%) and each of the isolates showed multiple antimicrobial resistance phenotype (resistant to ≥3 classes of antimicrobials). PCR amplification of antimicrobial resistance genes revealed that the plasmid-mediated quinolone resistance gene, qnrS, was the most prevalent (73.85%) among the isolates. The other antimicrobial resistance genes were detected in the following proportions: qnrB (26.15%), aac(6′)-Ib-cr (4.60%), tetA (16.92%), tetE (21.54%), aac(6′)-Ib (29.23%), and aphAI-IAB (7.69%). The IntI gene was found in 64.62% isolates, and four class 1 integron gene cassette profiles (incomplete dfrA1, catB3-aadA1, dfrA1-orfC, and qacE2-orfD) were identified. These data suggest that goldfish-borne Aeromonas spp. serve as a reservoir of antimicrobial resistance genes and class 1 integrons.
Get full access to this article
View all access options for this article.
References
1.
WinfreeR.A.1989. Tropical fish: their production and marketing in the United States. World Aquacult., 20:24–30.
2.
KrishnakumarK., RaghavanR., PrasadG., BijukumarA., SekharanM., PereiraB., and AliA.. 2009. When pets become pests–exotic aquarium fishes and biological invasions in Kerala. India. Curr. Sci., 97:474–476.
3.
JohnN., and HathaA.A.M.. 2013. Distribution, extracellular virulence factors and drug resistance of motile aeromonads in fresh water ornamental fishes and associated carriage water. Int. J. Fish Aquacult., 3:92–100.
4.
AustinB., and AustinD.A.. 1993. Bacterial fish pathogens. In SchusterS. (ed.), Diseases of farmed and wild fish. Springer Publication, Chicester, United Kingdom, pp.111–117.
5.
JandaJ.M., and AbbottS.L.. 2010. The genus Aeromonas: taxonomy, pathogenicity, and infection. Clin. Microbiol. Rev., 23:35–73.
6.
AustinD., McIntoshD., and AustinB.. 1989. Taxonomy of fish associated Aeromonas spp. with the description of Aeromonas salmonicida subsp. smithia. Syst. Appl. Microbiol., 11:277–290.
7.
DobiasovaH., KutilovaI., and PiackovaV.. 2014. Ornamental fish as a source of plasmid-mediated quinolone resistance genes and antibiotic resistance plasmids. Vet. Microbiol., 171:413–421.
8.
WeirM., RajićA., DutilL., UhlandC., and BruneauN.. 2012. Zoonotic bacteria and antimicrobial resistance in aquaculture: opportunities for surveillance in Canada. Can. Vet. J., 53:619–622.
9.
DiasC., MotaV., Martinez-MurciaA., and SaavedraM.J.. 2012. Antimicrobial resistance patterns of Aeromonas spp. isolated from ornamental fish. J. Aquacult. Res. Dev., 3:131.
10.
YangY., MiaoP., LiH., TanS., YuH., and YuH.. 2017. Antibiotic susceptibility and molecular characterization of Aeromonas hydrophila from grass carp. J. Food Saf.e12393.
11.
WeiL.S., MustakimM.T., AzlinaI.N., ZulhisyamA.K., AnamtM.N., WeeW., and HuangN.M.. 2015. Antibiotic and heavy metal resistance of Aeromonas spp. isolated from diseased red hybrid tilapia (Oreochromis sp.). Annu. Res. Rev. Biol., 6:264–269.
12.
LeeS.W., and WendyW.. 2017. Antibiotic and heavy metal resistance of Aeromonas hydrophila and Edwardsiella tarda isolated from red hybrid tilapia (Oreochromis spp.) coinfected with motile Aeromonas septicemia and edwardsiellosis. Vet. World, 10:803–807.
13.
El-BarbaryM.I.2017. Serum biochemical and histopathological changes associated with Aeromonas hydrophila isolated from Oreochromis niloticus and Sparus aurata with multiple antibiotic resistance index. J. Biol. Sci., 17:222–234.
14.
YanoY., HamanoK., TsutsuiI., Aue-umneoyD., BanM., and SatomiM.. 2015. Occurrence, molecular characterization, and antimicrobial susceptibility of Aeromonas spp. in marine species of shrimps cultured at inland low salinity ponds. Food Microbiol., 47:21–27.
15.
OdeyemiO.A., and AhmedA.. 2017. Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources. Saudi J. Biol. Sci., 24:65–70.
16.
Aravena-RománM., InglisT.J.J., HendersonB., RileyT.V., and ChangB.J.. 2012. Antimicrobial susceptibilities of Aeromonas strains isolated from clinical and environmental sources to 26 antimicrobial agents. Antimicrob. Agents Chemother., 56:1110–1112.
17.
WimalasenaS.H.M.P., De SilvaB.C.J., HossainS., PathiranaH.N.K.S., and HeoG.J.. 2017. Prevalence and characterization of quinolone resistance genes in Aeromonas species isolated from pet turtle in Korea. J. Glob. Antimicrob. Resist., 11:34–38.
18.
Evangelista-BarretoN.S., De CarvalhoF.C.T., VieiraR.H.S.S.F., dos ReisC.M.F., MacraeA., and RodriguesD.D.P.. 2010. Characterization of Aeromonas species isolated from an estuarine environment. Braz. J. Microbiol. 41:452–460.
19.
BoerlinP., and Reid-SmithR.J.. 2008. Antimicrobial resistance: its emergence and transmission. Anim. Health Res. Rev., 9:115–126.
20.
MazelD.2008. Integrons: agents of bacterial evolution. Nat. Rev. Microbiol., 4:608–620.
21.
DengY., BaoX., JiL., ChenL., LiuJ., MiaoJ., DingqiangC., HuaweiB., and YuG.. 2015. Resistance integrons: class 1, 2 and 3 integrons. Ann. Clin. Microbiol. Antimicrob., 14:45.
22.
DengY., WuY., JiangL., TanA., ZhangR., and LuoL.. 2016. Multi-drug resistance mediated by class 1 integrons in Aeromonas isolated from farmed freshwater animals. Front. Microbiol., 7:935.
23.
YáñezM.A., CatalánV., ApráizD., FiguerasM.J., and Martínez-MurciaA.J.. 2003. Phylogenetic analysis of members of the genus Aeromonas based on gyrB gene sequences. Int. J. Syst. Evol. Microbiol., 53:875–883.
24.
Clinical and Laboratory Standards Institute. 2014. Performance standards for antimicrobial susceptibility testing of bacteria isolated from aquatic animals. Second informational supplement. CLSI document VET03/VET04-S2. Clinical and Laboratory Standards Institute, Wayne, PA.
25.
ChungT., KimB., KimW., and ShinG.W.. 2017. Identification and antibiotic resistance profiling of bacterial isolates from septicaemic soft-shelled turtles (Pelodiscus sinensis). Vet. Med., 62:169–177.
26.
HossainS., De SilvaB.C.J., WimalasenaS.H.M.P., PathiranaH.N.K.S., and HeoG.-J.. 2017. Aminoglycoside susceptibility and genetic characterization of Salmonella enterica subsp. enterica isolated from pet turtles. Korean J. Vet. Serv., 40:27–33.
27.
SundeM., and SørumH.. 1999. Characterization of integrons in Escherichia coli of the normal intestinal flora of swine. Microb. Drug Resist., 5:279–287.
28.
KimD.-Y.2015. Fostering direction of the ornamental fish industry in Korea through a competitive analysis of international ornamental fish industry. J. Fish Bus. Adm., 46:15–28.
29.
TaylerA.E., AyalaJ.A., NiumsupP., WestphalK., BakerJ.A., ZhangL., WalshT.R., WiedemannB., BennettP.M., and AvisonM.B.. 2010. Induction of beta-lactamase production in Aeromonas hydrophila is responsive to beta-lactam-mediated changes in peptidoglycan composition. Microbiology, 156:2327–2335.
Verner-JeffreysD.W., WelchT.J., SchwarzT., PondM.J., WoodwardM.J., SarahJ.H., GeorginaS.E.R., RobertsE., MorrisonV., and Baker-AustinC.. 2009. High prevalence of multidrug-tolerant bacteria and associated antimicrobial resistance genes isolated from ornamental fish and their carriage water. PLoS One, 4:e8388.
32.
AriasA., SeralC., GudeM.J., and CastilloF.J.. 2010. Molecular mechanisms of quinolone resistance in clinical isolates of Aeromonas caviae and Aeromonas veronii bv sobria. Int. Microbiol., 13:135–141.
33.
HuangX.Z., FryeJ.G., ChahineM.A., GlennL.M., AkeJ.A., SuW., NikolichM.P., and LeshoE.P.. 2012. Characteristics of plasmids in multi-drug-resistant enterobacteriaceae isolated during prospective surveillance of a newly opened hospital in Iraq. PLoS One, 7:e40360.
34.
YuJ.E., ChoM.Y., KimJ., and KangH.Y.. 2012. Large antibiotic-resistance plasmid of Edwardsiella tarda contributes to virulence in fish. Microb. Pathog., 52:259–266.
35.
SchmidtA.S., BruunM.S., DalsgaardI., and LarsenJ.L.. 2001. Incidence, distribution and spread of tetracycline resistance determinants and integron-associated antibiotic resistance genes among motile aeromonads from a fish-farming environment. Appl. Environ. Microbiol., 67:5675–5682.
36.
NawazM., KhanA.A., KhanS., SungK., and SteeleR.. 2008. Isolation and characterization of tetracycline resistant Citrobacter spp. from catfish. Food Microbiol., 25:85–91.
37.
AkinbowaleO.L., PengH., and BartonM.D.. 2007. Diversity of tetracycline resistance genes in bacteria from aquaculture sources in Australia. J. Appl. Microbiol., 103:2016–2025.
38.
JagodaS.S.S.De S., HoneinK., ArulkanthanA., UshioH., and AsakawaS.. 2017. Genome sequencing and annotation of Aeromonas veronii strain Ae52, a multidrug-resistant isolate from septicaemic gold fish (Carassius auratus) in Sri Lanka. Genom. Data, 11:46–48.
39.
BarlowR.S., PembertonJ.M., DesmarchelierP.M., and GobiusK.S.. 2004. Isolation and characterization of integron-containing bacteria without antibiotic selection. Antimicrob. Agents Chemother., 48:838–842.
40.
BissonnetteL., and RoyP.H.. 1992. Characterization of In0 of Pseudomonas aeruginosa plasmid pVS1, an ancestor of integrons of multiresistance plasmids and transposons of gram-negative bacteria. J. Bacteriol., 174:1248–1257.
41.
ChangY.C., ShihD.Y., WangJ.Y., and YangS.S.. 2007. Molecular characterization of class 1 integrons and antimicrobials resistance in Aeromonas strains from foodborne outbreak-suspect samples and environmental sources in Taiwan. Diagn. Microbiol. Infect. Dis., 59:191–197.
42.
RoeM.T., VegaE., and PillaiS.D.. 2003. Antimicrobial resistance markers of class 1 and class 2 integron-bearing Escherichia coli from irrigation water and associated sediments. Emerg. Infect. Dis., 9:822–826.
43.
LevingsR.S., LightfootD., PartridgeS.R., HallR.M., and DjordjevicS.P.. 2005. The genomic island SGI1, containing the multiple antibiotic resistance region of Salmonella enterica serovar Typhimurium DT104 or variants of it, is widely distributed in other S. enterica serovars. J. Bacteriol., 187:4401–4409.
44.
ThungapathraM., Amita, SinhaK.K., ChaudhuriS.R., GargP., RamamurthyT., NairG.B., and GhoshA.. 2002. Occurrence of antibiotic resistance gene cassettes aac(6′)- Ib, dfrA5, dfrA12, and ereA2 in class I integrons in non-O1, non-O139 Vibrio cholerae strains in India. Antimicrob. Agents Chemother., 46:2948–2955.
45.
ShiL., FujiharaK., SatoT., ItoH., GargP., ChakrabartyR., RamamurthyT., NairG.B., TakedaY., and YamasakiS.. 2006. Distribution and characterization of integrons in various serogroups of Vibrio cholerae strains isolated from diarrhoeal patients between 1992 and 2000 in Kolkata, India. J. Med. Microbiol., 55:575–583.
46.
YuH.S., LeeJ.C., KangH.Y., RoD.W., ChungJ.Y., JeongY.S., TaeS.H., ChoiC.H., LeeE.Y., SeolS.Y., LeeY.C., and ChoD.T.. 2003. Changes in gene cassettes of class 1 integrons among Escherichia coli isolates from urine specimens collected in Korea during the last two decades. J. Clin. Microbiol., 41:5429–5433.
47.
JenningsM.C., MinbioleK.P., and WuestW.M.. 2015. Quaternary ammonium compounds: an antimicrobial mainstay and platform for innovation to address bacterial resistance. ACS Infect. Dis., 1:288–303.
