Aeromonas hydrophila IN FISH AND HUMANS; PREVALENCE, VIRULOTYPING AND ANTIMICROBIAL RESISTANCE

Authors

  • Heba A. Ahmed
  • Mohamed E.M. Mohamed
  • Maha M. Rezk
  • Rasha M. A. Gharieb
  • Shehata A. Abdel-Maksoud

DOI:

https://doi.org/10.26873/SVR-636-2018

Abstract

This work aimed to investigate the virulence factors and antibiotic resistance of A. hydrophila isolated from different sources in Damietta Governorate, Egypt. The samples comprised of tilapia (Oreochromis niloticus, n=150), mugil (Mugil cephalus, n=144), human stool (n=102) and fish sellers hand swabs (n=85). A total of 25 A. hydrophila isolates were recovered and molecularly confirmed, of which, 2.7% were from tilapia muscles, 2% from tilapia viscera, 6.3% from mugil viscera and 8.8% from stool samples. No isolates were recovered from mugil muscles and hand swabs. Phenotypic determination of heamolysis, lipolysis, proteolysis and gelatinase activity revealed that 60%, 56%, 60% and 100% were positive, respectively. Molecular identification of haemolysin (hly), aerolysin (aer), lipase (lip) and cytotoxic enterotoxins (act) virulence associated genes showed that 28%, 68%, 28% and 16% of the examined isolates were positive, respectively. Antimicrobial susceptibility of the isolates to 17 antibiotics was determined, the most resistance pattern was observed for cefixime (88%), while, all of the isolates were susceptible to imipenem. This study revealed that A. hydrophila isolated from fish and humans showed several virulence factors and exhibit a wide range of antibiotic resistance which is considered a public health hazard as well as.

Key words: Aeromonas hydrophila; virulence; vntibiotic resistance; fish

References

(1) FAO. Food and Agriculture Organization of the United Nations. The state of world fisheries and aquaculture Rome. 2009: 176.

(2) Citarasu T, Dhas AK, Velmurugan S, Viji TV, Kumaran T, Babu MM, Selvaraj T. Isolation of Aeromonas hydrophila from infected ornamental fish hatchery duriing massive outbreaks. Int J Curr Res. 2011; 2: 37–41.

(3) Sarkar A, Saha M, Roy P. Detection of 232bp Virulent Gene of Pathogenic Aeromonas hydrophilathrough PCR Based Technique: (A Rapid Molecular Diagnostic Approach). Adv Microbiol. 2013; 03(01): 83–7.

(4) Yogananth N, Bhakyaraj R, Chanthuru a, Anbalagan T, Nila KM. Detection of Virulence Gene in Aeromonas hydrophila Isolated from Fish Samples Using PCR Technique. Global J Biotech Biochem. 2009; 4(1): 51–3.

(5) Abbott SL, Cheung WK, Janda JM. The genus Aeromonas: biochemical characteristics, atypical reactions, and phenotypic identification schemes. J Clin Microbiol. 2003; 41(6): 2348–57.

(6) Martin-Carnahan A, Joseph SW. Genus I. Aeromonas. In: D.J. Brenner NRKaJTS, eds., editor. In Bergey’s Manual of Systematic Bacteriology. 2. New York, NY: Springer; 2005.

(7) Salunke G, Namshikar V, Gaonkar R, Gaonkar T. A case of Aeromonas hydrophila meningitis in septic shock. Trop J Med Res. 2015; 18(1): 54–.

(8) Austin B, Allen-Austin D. A Review: Bacterial pathogens of fish. J Appl Bacteriol. 1985; 58(5): 483–506.

(9) Borchardt MA, Stemper ME, Standridge JH. Aeromonas isolates from human diarrheic stool and groundwater compared by pulsed-field gel electrophoresis. Emerg Infect Dis. 2003; 9(2): 224–8.

(10) Tahoun AB, Ahmed HA, Abou Elez RM, El-Gedawy AA, Elsohaby I, Abd El-Ghafar AE. Molecular characterisation, genotyping and survival of Aeromonas hydrophila isolated from milk, dairy products and humans in Egypt. Int Dairy J. 2016; 63: 52–8.

(11) Sreeremya S. Identification , Characte-rization, Antibiotic Resistance of Aeromonas hydrophila in Chicken Intestine. IRE Journals. 2017; 1(3): 19–22.

(12) Wamala SP, Mugimba KK, Mutoloki S, Evensen Ø, Mdegela R, Byarugaba DK, Sørum H. Occurrence and antibiotic susceptibility of fish bacteria isolated from Oreochromis niloticus (Nile tilapia) and Clarias gariepinus (African catfish) in Uganda. Fish Aquatic Sci. 2018; 21 (1): 6.

(13) Umesha D, Srinivasa Rao P, Pani Prasad K, Reddy AK, Srinivas KN. Aerolysin and haemolysin virulence genes of Aeromonas hydrophila isolated from diseased ornamental freshwater Oscar fish and goldfish by Polymerase Chain Reaction. Int J Adv Sci Technol. 2011; 3 (1): 82–9.

(14) Kozaki S, Asao T, Kamata Y, Sakaguchi G. Characterization of Aeromonas sobria hemolysin by use of monoclonal antibodies against Aeromonas hydrophila hemolysins. J Clin Microbiol. 1989; 27 (8): 1782–6.

(15) Austin DA, McIntosh D, Austin B. Taxonomy of Fish Associated Aeromonas spp., with the Description of Aeromonas salmonicida subsp. smithia subsp. nov. Syst Appl Microbiol. 1989; 11 (3): 277–90.

(16) Martinez-Murcia AJ, Esteve C, Garay E, Collins MD. Aeromonas allosaccharophila sp. nov., a new mesophilic member of the genus Aeromonas. FEMS Microbiol Lett. 1992; 91 (3): 199–205.

(17) Odeyemi O, Asmat A, Usup G. Antibiotics resistance and putative virulence factors of Aeromonas hydrophila isolated from estuary. J Microbiol Biotechnol Food Sci. 2012; 1 (6): 1339.

(18) Sharma I, Rabha D, Das S, Ningombam D. Hemolytic activity and antibiotic resistance of Aeromonas sp. isolated from marketed fish. Eur J Pharm Med Res 2015; 2 (4), 304–312.

(19) Odeyemi OA, Ahmad A. Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources. Saudi J Bio Sci. 2017; 24 (1): 65–70.

(20) Didugu H, Krishnaiah N , Rao MT, Ramanipushpa RN . Antibiogram of Aeromonas Species Isolated From Livestock Products. Int J Sci Environ Technol. 2016; 5 (2): 475–8.

(21) ICMSF. International commission on Mic-robial specification for foods. Microorganisms in foods. Microbiol Ecology and Food Commodities London, IK: Blackie Academic and Professional. 1998.

(22) Austin B, Austin DA. Bacterial Fish Pathogens: Disease in Farmed and Wild Fish. Chichester: Praxis Publishing Ltd; 1999.

(23) MacFaddin JF. Biochemical tests for identification medical bacteria. Baltimore, Md. 21202 USA: Warery Press Inc; 2000.

(24) Gordon L, Giraud E, Ganière JP, Armand F, Bouju-Albert A, De La Cotte N, Mangion C, Le Bris H. Antimicrobial resistance survey in a river receiving effluents from freshwater fish farms. J Appl Microbiol. 2007; 102 (4): 1167–76.

(25) Singh DV, Sanyal SC. Production of hae-molysis and its correlation with enterotoxicity in Aeromonas spp. J Med Microbiol. 1992; 37 (4): 262–7.

(26) Duza MB, Mastan S. Optimization of lipase production from Bacillus thurngiensis (Ts11BP), Achromobacter xylosoxidans J2 (Ts2 MCN)-isolated from soil sediments near oilseed farm. IOSR j Pharm Biol Sci. 2014; 9: 66–76.

(27) Rivero O, Anguita J, Paniagua C, Naharro G. Molecular cloning and characterization of an extracellular protease gene from Aeromonas hydrophila. J Bacteriol 1990; 172 (7): 3905–8.

(28) Olaniran AO, Nzimande SBT, Mkize NG. Antimicrobial resistance and virulence signatures of Listeria and Aeromonas species recovered from treated wastewater effluent and receiving surface water in Durban, South Africa. BMC Microbiol 2015; 15 (1): 1–10.

(29) Yousr AH, Napis S, Rusul GRA, Son R. Detection of aerolysin and hemolysin genes in Aeromonas spp. Isolated from environmental and shellfish sources by polymerase chain reaction. IntFood Res J 2007; 14 (2): 115–22.

(30) Singh V, Rathore G, Kapoor D, Mishra BN, Lakra WS. Detection of aerolysin gene in Aeromonas hydrophila isolated from fish and pond water. Indian J Microbiol 2008; 48 (4): 453–8.

(31) Nawaz M, Khan SA, Khan AA, Sung K, Tran Q, Kerdahi K, Steele R. Detection and characterization of virulence genes and integrons in Aeromonas veronii isolated from catfish. Food Microbiol 2010; 27 (3): 327–31.

(32) Sen K, Rodgers M. Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities: a PCR identi-fication. J Appl Microbiol 2004; 97 (5): 1077–86.

(33) CLSI. Clinical and Laboratory Standards Institute. Methods for Antimicrobial Dilution and Disk Susceptibility Testing of Infrequently Isolated or Fastidious Bacteria; approved guideline M45-P. Proposed Guideline. CLSI document M45-P. 2006; Vol. 25, No. 26.

(34) EUCAST. The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 8, 2018. http: //www.eucast.org. 2018.

(35) CLSI. Clinical and Laboratory Standards Institute. M100 Performance Standards for Antimicrobial Susceptibility Testing 2017. PP: 296.

(36) CASFM/EUCAST. Enterobacteriaceae. In: French Society of Microbiology Ed; 2018, pp: 36-44. 2018.

(37) CASFM/EUCAST. Aeromonas sp. In: French Society of Microbiology, Ed; 2018, p: 117. 2018.

(38) CASFM. Comite´ de l’Antibiogramme de la Socie´te´ Franc¸aise de Microbiologie; proposer les recommandations 2013. 2013.

(39) Krumperman PH. Multiple Antibiotic Res-istance Indexing of Escherichia coli to Identify High-Risk Sources of Fecal Contamination of Foo-dst. Appl Environ Microbiol. 1983; 46 (1):165–70.

(40) Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, Harbarth S, Hindler JF, Kahlmeter G, Olsson-Liljequist B, Paterson DL, Rice LB, Stelling J, Struelens MJ, Vatopoulos A, Weber JT, Monnet DL. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resis-tance. Clin Microbiol Infect. 2012; 18 (3): 268–81.

(41) Arslan S, Küçüksari R. Phenotypic and Genotypic Virulence Factors and Antimicrobial Resistance of Motile Aeromonas spp. from Fish and Ground Beef. J Food Saf. 2015; 35 (4): 551–9.

(42) Ibrahem MD, Mostafa MM, Arab RM, Rezk MA, editors. Prevalence of Aeromonas hydrophila infection in wild and cultured tilapia nilotica (O. niloticus) in Egypt. Proceedings of 8th International Symposium on Tilapia in Aquaculture; 2008; Cairo, Egypt. pp. 1257–71

(43) Ashiru AW, Uaboi-Egbeni PO , Oguntowo JE, Idika CN. Isolation and antibiotic profile of Aeromonas species from tilapia and catfish fish.pdf. Pak J Nutr 2011; 10 (10): 982–6.

(44) Castro-Escarpulli G, Figueras MJ, Aguilera-Arreola G, Soler L, Fernández-Rendón E, Aparicio GO, Guarro J, Chacón MR. Characterisation of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico. Int J Food Microbiol. 2003; 84 (1): 41–9.

(45) Ibrahim HM, Amin RA, Shawkey NA, Mohammed HE. Bacteriological Evaluation of Some Fresh and Frozen fish. Benha Vet Med J. 2016; 31 (1): 24–9

(46) Ramadan H, Ibrahim N, Samir M, Abd El-Moaty A, Gad T. Aeromonas hydrophila from marketed mullet (Mugil cephalus) in Egypt: PCR characterization of β-lactam resistance and virulence genes. J Appl Microbiol. 2018; 124 (6): 1629–37.

(47) Cipriano RC, Bullock GL, Pyle S. Aeromonas hydrophila and motile aeromonad septicemias of fish. 1984.

(48) Subashkumar R, Thayumanavan T, Vivekanandhan G, Lakshmanaperumalsamy P. Occurrence of Aeromonas hydrophila in acute gas-teroenteritis among children. Indian J Med Res. 2006; 123 (1): 61–6.

(49) Mohana SJ. Sequestration of Aeromonas hydrophila for serine protease gene detection from diarrhoeal samples through pcr. Innovative Journal of Medical and Health Science. 2015; 5 (5). 2015. 210–8 p.

(50) Mohammed EAM. The role of fish and other invertebrate aquatic organisms in transmitting some bacterial diseases to man: MVSc thesis, Cairo University; 2012.

(51) Rahim Z, Sanyal SC, Aziz KM, Huq MI, Chowdhury AA. Isolation of enterotoxigenic, hemolytic, and antibiotic-resistant Aeromonas hydrophila strains from infected fish in Bangladesh. Appl Environ Microbiol. 1984; 48 (4): 865–7.

(52) Bhowmik P, Bag PK, Hajra TK, De R, Sarkar P, Ramamurthy T. Pathogenic potential of Aeromonas hydrophila isolated from surface waters in Kolkata, India. J Med Microbiol. 2009; 58 (12): 1549–58.

(53) Simon SS, Lalitha KV, Joseph TC. Virulence properties of Aeromonas spp. from modified-atmosphere- and vacuum-packed milk fish (Chanos chanos Forsskal, 1775). Ann Microbiol. 2016; 66 (3): 1109–15.

(54) Tom ás JM. The Main Aeromonas Patho-genic Factors. ISRN Microbiol. 2012; 2012: 22.

(55) Ye YW, Fan TF, Li H, Lu JF, Jiang H, Hu W, Jiang QH. Characterization of Aeromonas hydrophila from hemorrhagic diseased freshwater fishes in Anhui Province, China. Int Food Res J 2013; 20 (3): 1449–52.

(56) Younes AM, Gaafar AY, Awad ES. Virulence Determinants and Plasmid Profile of Aeromonas hydrophila Strains Isolated from Oreo-chromis niloticus. Global Vet 2015; 15 (6): 613–7.

(57) Sarder H, Khan T, Saha ML, Punom NJ, Mandal SC, Rahman MS. Prevalence and antibiotic susceptibility of Aeromonas hydrophila isolated from freshwater fishes. J Fish 2016; 4 (3): 411–9.

(58) TopićPopovic N, Teskeredzić E, Strunjak-Perovic I, Čož-Rakovac R. Aeromonas hydrophila isolated from wild freshwater fish in Croatia. Vet Res Commun. 2000; 24 (6): 371–7.

(59) Hatha M, Vivekanandhan AA, Joice GJ, Christol. Antibiotic resistance pattern of motile aeromonads from farm raised fresh water fish. Int J Food Microbiol. 2005; 98 (2): 131–4.

(60) Sreedharan K, Philip R, Singh ISB. Virulence potential and antibiotic susceptibility pattern of motile aeromonads associated with freshwater ornamental fish culture systems: A possible threat to public health. Braz J Microbiol. 2012; 43 (2): 754–65.

(61) Rathore G, Swaminathan TR, Mahanta PC, Kapoor D. Isolation and characterization of motile aeromonads from aquatic environment. IJFAS 2005; 52 (2): 241–8.

(62) McDermott PF, Zhao S, Wagner DD, Simjee S, Walker RD, White DG. The food safety perspective of antibiotic resistance. Anim Biotechnol. 2002; 13 (1): 71–84.

Downloads

Published

2018-11-06

Issue

Section

Original research Article