Veterinary World

     Open access and peer reviewed journal  

ISSN (Online): 2231-0916


Home l Editorial board l Instructions for authors l Reviewer guideline l Open access policy l Archives l FAQ

Open Access

Copyright: The authors. This article is an open access article licensed under the terms of the Creative Commons Attribution License

( which permits unrestricted use, distribution and reproduction in any medium, provided the work is properly cited.

Research (Published online: 28-10-2015)

14. A study on the prevalence of Aeromonas spp. and its enterotoxin genes in samples of well water, tap water, and bottled water - Hareesh Didugu, Madhavarao Thirtham, Krishnaiah Nelapati, K Kondal Reddy, Baba Saheb Kumbhar, Anusha Poluru and Guruvishnu Pothanaboyina

Veterinary World, 8(10): 1237-1242



   doi: 10.14202/vetworld.2015.1237-1242


Hareesh Didugu: Animal Disease Diagnostic Laboratory, Vijayawada, Andhra Pradesh, India;

Madhavarao Thirtham: Department of Veterinary Public Health and Epidemiology, College of Veterinary Science, Proddatur, Andhra Pradesh, India;

Krishnaiah Nelapati: Department of Veterinary Public Health, College of Veterinary Science, Rajendranagar, Telangana, India;

K Kondal Reddy: Department of Livestock Products Technology, Sri P.V. Narsimha Rao Telangana State University for Veterinary, Animal and Fishery Sciences, Rajendranagar, Telangana, India;

Baba Saheb Kumbhar: Department of Veterinary Public Health, College of Veterinary Science, Rajendranagar, Telangana, India;

Anusha Poluru: Department of Veterinary Public Health, College of Veterinary Science, Rajendranagar, Telangana, India;

Guruvishnu Pothanaboyina: Department of Animal Genetics and Breeding, College of Veterinary Science, Proddatur, Andhra Pradesh, India;


Received: 07-07-2015, Revised: 11-09-2015, Accepted: 18-09-2015, Published online: 28-10-2015


Corresponding author: Hareesh Didugu, e-mail:

Citation: Didugu H, Thirtham M, Nelapati K, Reddy KK, Kumbhar BS, Poluru A, Pothanaboyina G (2015) A study on the prevalence of Aeromonas spp. and its enterotoxin genes in samples of well water, tap water, and bottled water, Veterinary World 8(10): 1237-1242.

Aim: The aim of this work was to study the prevalence of Aeromonas spp. and its enterotoxin genes in various water sources.

Materials and Methods: 125 samples (50 from well water, 50 from tap water, and 25 from bottled water) were collected from various sources in and around Greater Hyderabad Municipal Corporation and examined for the presence of aeromonads by both cultural and polymerase chain reaction (PCR) assay. Alkaline peptone water with ampicillin was used as enrichment. Aeromonas isolation medium and ampicillin dextrin agar were used as selective media. The boiling and snap chilling method was used for DNA extraction. Primers targeted against 16S rRNA, aer, and ast were used to identify aeromonads and its enterotoxins.

Results: 48%, 18%, and 12% of well water, tap water, and bottled water samples were found positive by cultural assay with an overall prevalence of 28.8%. Aeromonads were detected in 32 % (52% in well water, 20% in tap water, and 16% in bottled water) of samples by PCR assay. Aerolysin (aer) gene was noticed in 34.6%, 20%, and 0% of well water, tap water, and bottled water samples, respectively, with an overall prevalence of 27.5%. Thermostable cytotonic enterotoxin (ast) was observed in 37.5% (42.3% in well water, 30% in tap water, and 25% in bottled mineral water) of samples.

Conclusions: Presence of aeromonads and its toxin genes in various sources of water is of public health concern and emphasizes the need for necessary preventive measures to tackle the problem.

Keywords: Aeromonas spp., enterotoxins, polymerase chain reaction, prevalence, water.

1. Pandove, G., Sahota, P., Kumar, S. and Jairath, S. (2013) Aeromonas hydrophila: An emerging pathogen in drinking water. Int. J. Health Pharm. Sci., 2(1): 15-26.
2. Igbinosa, I.H., Igumbor, E.U., Aghdasi, F., Tom, M. and Okoh, A.I. (2012) Emerging aeromonas species infections and their significance in public health. Sci. World J., 2012: Article ID:625023.
3. Robertson, B.K., Harden, C., Selvaraju, S.B., Pradhan, S. and Yadav, J.S. (2014) Molecular detection, quantification, and toxigenicity profiling of Aeromonas spp. in source-and drinking-water. Open Microbiol. J., 8: 32.
PMid:24949108 PMCid:PMC4062929
4. Hasan, J. (2006) Aeromonas: Human health criteria document. EPA Office Water, United States.
5. WHO. (2011) Guidelines for Drinking-Water Quality. World Health Organization, Geneva.
6. Sha, J., Kozlova, E.V. and Chopra, A.K. (2002) Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: Generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity. Infect. Immun., 70(4): 1924-1935.
7. Janda, J.M. and Abbott, S.L. (2010) The genus Aeromonas: Taxonomy, pathogenicity, and infection. Clin. Microbiol. Rev., 23(1): 35-73.
PMid:20065325 PMCid:PMC2806660
8. Khajanchi, B.K., Fadl, A.A., Borchardt, M.A., Berg, R.L., Horneman, A.J., Stemper, M.E., Joseph, S.W., Moyer, N.P., Sha, J. and Chopra, A.K. (2010) Distribution of virulence factors and molecular fingerprinting of Aeromonas species isolates from water and clinical samples: Suggestive evidence of water-to-human transmission. Appl. Environ. Microbiol., 76(7): 2313-2325.
PMid:20154106 PMCid:PMC2849238
9. Venkataiah, P., Poojary, N.S. and Harshvardhan, B. (2013) A multiplex PCR for detection of haemolytic Aeromonas hydrophila from vegetable sources in Karnataka, India. Recent Res. Sci. Technol., 5(3): 19-23.
10. Yousr, A.H., Napis, S., Ali, R., Rusul, G. and Radu, S. (2007) Detection of aerolysin and hemolysin genes in Aeromonas spp. isolated from environmental and shellfish sources by polymerase chain reaction. ASEAN Food J., 14(2): 115-122.
11. Arora, S., Agarwal, R.K. and Bist, B. (2006) Comparison of ELISA and PCR vis-ŕ-vis cultural methods for detecting Aeromonas spp. in foods of animal origin. Int. J. Food Microbiol., 106(2): 177-183.
12. Sen, K. and Rodgers, M. (2004). Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities: A PCR identification. J. Appl. Microbiol., 97(5): 1077-1086.
13. Ghenghesh, K.S., Ahmed, S.F., El-Khalek, R.A., Al-Gendy, A. and Klena, J. (2008) Aeromonas-associated infections in developing countries. J. Infect. Dev. Ctries., 2(2): 081-098.
14. Elmanama, A.A. and Ferwana, N. (2011) Yersinia enterocolitica and Aeromonas hydrophila in clinical, food and environmental samples in gaza strip. J. Al Azhar Univ. Gaza., 13: 69-82.
15. Scoaris, D.D.O., Bizerra, F.C., Yamada-Ogatta, S.F., Abreu Filho, B.A.D., Ueda-Nakamura, T., Nakamura, C.V. and Dias Filho, B.P. (2008) The occurrence of Aeromonas spp. In the bottled mineral water, well water and tap water from the municipal supplies. Braz. Arch. Biol. Technol., 51(5): 1049-1055.
16. Aguilera Arreola, M., Hernández Rodríguez, C., Zú-iga, G., Figueras, M.J. and Castro Escarpulli, G. (2005) Aeromonas hydrophila clinical and environmental ecotypes as revealed by genetic diversity and virulence genes. FEMS Microbiol. Lett., 242(2): 231-240.
17. Řrmen, Ř. and Řstensvik, Ř. (2001) The occurrence of aerolysin‐positive Aeromonas spp. and their cytotoxicity in Norwegian water sources. J. Appl. Microbiol., 90(5): 797-802.
18. Gibotti, A., Saridakis, H.O., Pelayo, J.S., Tagliari, K.C. and Falcao, D.P. (2000) Prevalence and virulence properties of Vibrio cholerae non‐O1, Aeromonas spp. and Plesiomonas shigelloides isolated from Cambé Stream (State of Paraná, Brazil). J. Appl. Microbiol., 89(1): 70-75.
19. Eid, H.M., Shalaby, A.M. and Soltan, S.E.S. (2013) Incidence of Aeromonas species isolated from different food sources and water. Available from:
20. Pablos, M., Rodríguez-Calleja, J.M., Santos, J.A., Otero, A. and García-López, M.L. (2009) Occurrence of motile Aeromonas in municipal drinking water and distribution of genes encoding virulence factors. Int. J Food Microbiol., 135(2): 158-164.
21. Balakrishna, K., Murali, H.S. and Batra, H.V. (2010) Detection of toxigenic strains of Aeromonas species in foods by a multiplex PCR assay. Indian J. Microbiol., 50(2): 139-144.
PMid:23100820 PMCid:PMC3450327
22. Emekdas, G., Aslan, G., Tezcan, S., Serin, M.S., Yildiz, C., Ozturhan, H. and Durmaz, R. (2006) Detection of the frequency, antimicrobial susceptibility, and genotypic discrimination of Aeromonas strains isolated from municipally treated tap water samples by cultivation and AP-PCR. Int. J. Food Microbiol., 107(3): 310-314.
23. Alavandi, S.V. and Ananthan, S. (2003) Biochemical characteristics, serogroups, and virulence factors of Aeromonas species isolated from cases of diarrhoea and domestic water samples in Chennai. Indian J. Med. Microbiol., 21(4): 233.
24. Fernández, M.C., Giampaolo, B.N., Iba-ez, S.B., Guagliardo, M.V., Esnaola, M.M., Conca, L., Valdivia, P., Stagnaro, S.M., Chiale, C. and Frade, H. (2000) Aeromonas hydrophila and its relation with drinking water indicators of microbiological quality in argentine. Genetica, 108(1): 35-40.
25. Ivanova, E.P., Zhukova, N.V., Gorshkova, N.M. and Chaikina, E.L. (2001) Characterization of Aeromonas and Vibrio species isolated from a drinking water reservoir. J. Appl. Microbiol., 90(6): 919-927.
26. Bhowmik, P., Bag, P.K., Hajra, T.K., De, R., Sarkar, P. and Ramamurthy, T. (2009) Pathogenic potential of Aeromonas hydrophila isolated from surface waters in Kolkata, Indian. J. Med. Microbiol., 58(12): 1549-1558.
27. Biscardi, D., Castaldo, A., Gualillo, O. and De Fusco, R. (2002) The occurrence of cytotoxic Aeromonas hydrophila strains in Italian mineral and thermal waters. Sci. Total Environ., 292(3): 255-263.
28. Slade, P.J., Falah, M.A. and Al-Ghady, A.M. (1986), Isolation of Aeromonas hydrophila from bottled waters and domestic water supplies in Saudi Arabia. J. Food Protect., 49(6): 471-476.
29. Erdem, A.K., Gürün, S., Zeybek, Z., Dogruoz, N. and Cotuk, A. (2009) Microbiological investigation of bottled waters from different suppliers from Istanbul. IUFS J. Biol., 68(1): 37-45.
30. Agarwal, R.K., Kapoor, K.N., Kumar, A. and Bhilegaonkar, K.N. (2000) Aeromonads in foods of animal origin. Indian J. Anim. Sci., 70(9): 942-943.
31. Gugliandolo, C., Lentini, V., Spanň, A. and Maugeri, T.L. (2011). Conventional and molecular methods to detect bacterial pathogens in mussels. Lett. Appl. Microbiol., 52(1): 15-21.
32. Liu, D., editor. (2011) Molecular Detection of Human Bacterial Pathogens. CRC Press, Boca Raton.
33. Onuk, E.E., Findik, A., Turk, N., Altun, S., Korun, J., Ozer, S., Avsever, M.L. and Ciftci, A. (2013) Molecular identification and determination of some virulence genes of Aeromonas spp. in fish and water from Turkish coastal regions. Rev. Méd. Vét., 164(4): 200-206.
34. Ottaviani, D., Parlani, C., Citterio, B., Masini, L., Leoni, F., Canonico, C., Sabatini, L., Bruscolini, F. and Pianetti, A. (2011) Putative virulence properties of Aeromonas strains isolated from food, environmental and clinical sources in Italy: A comparative study. Int. J. Food Microbiol., 144(3): 538-545.
35. Balsalobre, L., Dropa, M., Matte, G. and Matte, M. (2009) Molecular detection of enterotoxins in environmental strains of Aeromonas hydrophila and Aeromonas jandaei. J. Water. Health, 7(4): 685-691.
36. Seshadri, R., Joseph, S.W., Chopra, A.K., Sha, J., Shaw, J., Graf, J. and Heidelberg, J.F. (2006) Genome sequence of Aeromonas hydrophila ATCC 7966T: Jack of all trades. J. Bacteriol., 188(23): 8272-8282.
PMid:16980456 PMCid:PMC1698176