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Identification and characterisation of emerging fish pathogens Aeromonas veronii and Aeromonas hydrophila isolated from naturally infected Channa punctata

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Abstract

Naturally infected Channa punctata exhibiting bacterial septicemic syndrome including ulcerations along with mortality records were collected from a fish farm in Assam during winter season (early November 2020 to early January 2021). The moribund fishes were subjected for bacterial isolation followed by identification of the bacteria. Two dominant emerging bacterial pathogens were identified as Aeromonas veronii (isolate ZooGURD-01) and Aeromonas hydrophila (isolate ZooGURD-05) by standard biochemical characterization and 16S rRNA and rpo B gene amplification. Re-infection experiments of both the bacterial isolates in healthy disease-free C. punctata showed similar symptoms to that of natural infection thus confirming their virulence. The LD50 calculated during challenge test for both the isolates ZooGURD-01 and ZooGURD-05 found to be pathogenic at 2.6 × 104 and 1.6 × 104 CFU/fish respectively. Further PCR amplification of specific virulent genes (aerolysin, hemolysin and enterotoxin) confirmed pathogenicity for both isolates. Histopathological examinations of liver and kidney in re-infection experiments showed prominent changes supporting bacterial septicaemia. Antibiotic sensitivity pattern showed that the isolates ZooGURD-01 and ZooGURD-05 were sensitive to 22 and 19 out of 25 antimicrobials respectively. The present study was the first report on the mortality of farmed C. punctata associated with natural infection caused by A. veronii and A. hydrophila with no record of pathogenicity of A. veronii in C. punctata.

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Data availability

All data generated in the present study is provided in this article. Nucleotide sequences can be freely retrieved from the NCBI database (Accession Numbers: MZ400683, ON413743, ON073793 and ON458151).

References

  • Adhikary A et al (2022) Isolation, identification and characterisation of Aeromonas veronii from diseased Indian major carp, Cirrhinus mrigala. Aquac Int 31:997–1010

    Article  Google Scholar 

  • Albert MJ et al (2000) Prevalence of enterotoxin genes in Aeromonas spp. isolated from children with diarrhoea, healthy controls, and the environment. J Clin Microbiol 38:3785–3790

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Aly S, Eissa I, Badran A, Elamie M, Hussain B (2005) Pathological studies on encysted metacercariae infections among some freshwater fish in Egyptian aquaculture. Proc Duet Trop Hohenheim Univ Stuttg Ger 1:11–13

    Google Scholar 

  • Austin B, Austin DA (2016) Aeromonadaceae representatives (motile aeromonads). In: Austin B, Austin DA (eds) Bacterial fish pathogens, 1st edn. Springer, pp 161–214

    Chapter  Google Scholar 

  • Beaz-Hidalgo R et al (2015a) Aeromonas aquatica sp. nov., Aeromonas finlandiensis sp. nov. and Aeromonas lacus sp. nov. isolated from finnish waters associated with cyanobacterial blooms. Syst Appl Microbiol 38:161–168

    Article  CAS  PubMed  Google Scholar 

  • Beaz-Hidalgo R, Hossain MJ, Liles MR, Figueras MJ (2015b) Strategies to avoid wrongly labelled genomes using as example the detected wrong taxonomic affiliation for Aeromonas genomes in the GenBank database. PLoS ONE 10:e0115813

    Article  PubMed  PubMed Central  Google Scholar 

  • Carriero MM, Mendes Maia AA, Moro Sousa RL, Henrique-Silva F (2016) Characterization of a new strain of Aeromonas dhakensis isolated from diseased pacu fish (Piaractus mesopotamicus) in Brazil. J Fish Dis 39:1285–1295

    Article  CAS  PubMed  Google Scholar 

  • Chakrabarty NM (2006) Murrels and murrel culture. Narendra Pub House, India

    Google Scholar 

  • Chandrarathna HPSU et al (2018) Outcome of co-infection with opportunistic and multidrug resistant Aeromonas hydrophila and A. veronii in zebrafish: identification, characterization, pathogenicity and immune responses. Fish Shellfish Immunol 80:573–581

    Article  CAS  PubMed  Google Scholar 

  • Chen YF, Liang RS, Zhuo XL, Wu XT, Zou JX (2012) Isolation and characterization of Aeromonas schubertii from diseased snakehead, Channa maculata (Lacepède). J Fish Dis 35:421–430

    Article  CAS  PubMed  Google Scholar 

  • Chen F et al (2019) Isolation, identification and characteristics of Aeromonas veronii from diseased crucian carp (Carassius auratus gibelio). Front Microbiol 10:2742

    Article  PubMed  PubMed Central  Google Scholar 

  • Chu WH, Lu CP (2005) Multiplex PCR assay for the detection of pathogenic Aeromonas hydrophila. J Fish Dis 28:437–441

    Article  CAS  PubMed  Google Scholar 

  • Cipriano RC (1984) Aeromonas hydrophila and motile aeromonad septicemias of fish in fish disease leaflet 68 (US Department of the Interior, Fish and Wildlife Service, Division of Fishery Research), USA

  • Czerwińska-Główka D, Krukiewicz K (2021) Guidelines for a morphometric analysis of prokaryotic and eukaryotic cells by scanning electron microscopy. Cells 10:3304

    Article  PubMed  PubMed Central  Google Scholar 

  • Daskalov H (2006) The importance of Aeromonas hydrophila in food safety. Food Control 17:474–483

    Article  Google Scholar 

  • Dong HT et al (2017) Aeromonas jandaei and Aeromonas veronii caused disease and mortality in Nile tilapia, Oreochromis niloticus (L.). J Fish Dis 40:1395–1403

    Article  CAS  PubMed  Google Scholar 

  • Dorsch M, Ashbolt NJ, Cox PT, Goodman AE (1994) Rapid identification of Aeromonas species using 16S rDNA targeted oligonucleotide primers: a molecular approach based on screening of environmental isolates. J Appl Bacteriol 77:722–726

    Article  CAS  PubMed  Google Scholar 

  • Du Sert NP et al (2020) Reporting animal research: explanation and elaboration for the ARRIVE guidelines 2.0. PLoS Biol 18:pe3000411

    Article  Google Scholar 

  • Du Z et al (2020) A case of Aeromonas veronii infection in golden Taiwanese loach (Paramisgurnus dabryanus) at low water temperature. Aquac Res 51:1307–1312

    Article  Google Scholar 

  • El-Shahawy IS, El-Seify MO, Metwally AM, Fwaz MM (2017) Survey on endoparasitic fauna of some commercially important fishes of the river Nile, southern of Egypt (Egypt). Rev De Med Vet 168:126–134

    CAS  Google Scholar 

  • Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evol 39:783–791

    Article  Google Scholar 

  • Fernández-Bravo A, Figueras MJ (2020) An update on the genus Aeromonas: Taxonomy, epidemiology, and pathogenicity. Microorganisms 8:129

    Article  PubMed  PubMed Central  Google Scholar 

  • Finney DJ (1997) Probit analysis. Cambridge University Press, Cambridge

    Google Scholar 

  • Garcia LS, Procop GW (2016) Diagnostic medical parasitology. Manual of commercial methods in clinical microbiology, International. Wiley, pp 284–308

    Chapter  Google Scholar 

  • Hall TA (1999) Bio Edit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic acids symposium series number 41. Oxford University Press, Oxford, pp 95–98

    Google Scholar 

  • Haniffa MA, Marimuthu K, Nagarajan M, Arokiaraj AJ, Kumar D (2004) Breeding behaviour and parental care of the induced bred spotted murrel Channa punctatus under captivity. Curr Sci 86:1375–1376

    Google Scholar 

  • Hedge PJ, Spratt BG (1985) Resistance to β-lactam antibiotics by re-modelling the active site of an E. coli penicillin-binding protein. Nature 318:478–480

    Article  CAS  PubMed  Google Scholar 

  • Holt JG, Krieg NR, Sneath PH, Staley JT, Williams ST (1994) Bergey's manual of determinate bacteriology. William & Wilkins, Baltimore, MD, USA

  • Janda JM, Abbott SL (2010) The genus Aeromonas: taxonomy, pathogenicity, and infection. Clin Microbiol Rev 23:35–73

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jayaram KC (1981) Freshwater fishes of India, Pakistan, Bangladesh, Burma and Sri Lanka: a handbook, India

  • Khoo L (2000) Fungal diseases in fish. In: Ahjfdj A (ed) Seminars in Avian and exotic pet medicine, vol 9. WB Saunders, pp 102–111

    Google Scholar 

  • Kingombe CIB, Huys G, Tonolla M, Albert MJ, Swings J, Peduzzi R, Jemmi T (1999) PCR detection, characterization, and distribution of virulence genes in Aeromonas spp. Appl Environ Microbiol 65:5293–5302

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kozińska A, Paździor E, Pękala A, Niemczuk W (2014) Acinetobacter johnsonii and Acinetobacter lwoffii-the emerging fish pathogens. J Vet Res 58:193–199

    Google Scholar 

  • Kühn I et al (1997) Characterization of Aeromonas spp. isolated from humans with diarrhoea, from healthy controls, and from surface water in Bangladesh. J Clin Microbiol 35:369–373

    Article  PubMed  PubMed Central  Google Scholar 

  • Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Küpfer M, Kuhnert P, Korczak BM, Peduzzi R, Demarta A (2006) Genetic relationships of Aeromonas strains inferred from 16S rRNA, gyrB and rpoB gene sequences. Int J Syst Evol Microbiol 56:2743–2751

    Article  PubMed  Google Scholar 

  • Lazado CC, Zilberg D (2018) Pathogenic characteristics of Aeromonas veronii isolated from the liver of a diseased guppy (Poecilia reticulata). Lett Appl Microbiol 67:476–483

    Article  CAS  PubMed  Google Scholar 

  • Lewis WM, Bender M (1961) Free-living ability of a warm-water fish pathogen of the genus Aeromonas and factors contributing to its infection of the golden shiner. Prog Fish Cult 23:124–126

    Article  Google Scholar 

  • Liu C (2018) Aeromonas shuberti as a cause of multi-organ necrosis in internal organs of Nile tilapia, Oreochromis niloticus. J Fish Dis 41:1529–1538

    Article  CAS  PubMed  Google Scholar 

  • Luna LG (1968) Manual of histologic staining methods of the Armed Forces Institute of Pathology. McGraw-Hill, Blakiston Division

    Google Scholar 

  • Ma Z, Yang H, Li T, Luo L, Gao J (2009) Isolation and identification of pathogenic Aeromonas veronii isolated from infected Siberian sturgeon (Acipenser baerii). Acta Microbiol Sin 49:1289–1294

    CAS  Google Scholar 

  • Malick RC et al (2020) Identification and pathogenicity study of emerging fish pathogens Acinetobacter junii and Acinetobacter pittii recovered from a disease outbreak in Labeo catla (Hamilton, 1822) and Hypophthalmichthys molitrix (Valenciennes, 1844) of freshwater wetland in West Bengal, India. Aquac Res 51:2410–2420

    Article  CAS  Google Scholar 

  • Mallik SK et al (2020) Characterization and pathogenicity of Aeromonas veronii associated with mortality in cage farmed grass carp, Ctenopharyngodon idella (Valenciennes, 1844) from the Central Himalayan region of India. Antonie Van Leeuwenhoek 113:2063–2076

    Article  CAS  PubMed  Google Scholar 

  • Marinho-Neto FA et al (2019) Morphological, microbiological and ultrastructural aspects of sepsis by Aeromonas hydrophila in Piaractus mesopotamicus. PLoS ONE 14:e0222626

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martinez-Murcia AJ (2011) Multilocus phylogenetic analysis of the genus Aeromonas. Syst Appl Microbiol 34:189–199

    Article  CAS  PubMed  Google Scholar 

  • Mazumder A, Choudhury H, Dey A, Sarma D (2021) Isolation and characterization of two virulent Aeromonads associated with haemorrhagic septicaemia and tail-rot disease in farmed climbing perch Anabas testudineus. Sci Rep 11:1–10

    Article  Google Scholar 

  • Miñana-Galbis D, Farfán M, Lorén JG, Fusté MC (2002) Biochemical identification and numerical taxonomy of Aeromonas spp. isolated from environmental and clinical samples in Spain. J Appl Microbiol 93:420–430

    Article  PubMed  Google Scholar 

  • Mo J, Jiang L, Wu Z (2016) Biological characteristics and drug susceptibility of Aeromonas schubertii WL-4 isolated from snakehead. J Fish China 40:484–494

    Google Scholar 

  • Nagar V, Shashidhar R, Bandekar JR (2011) Prevalence, characterization, and antimicrobial resistance of Aeromonas strains from various retail food products in Mumbai, India. J Food Sci 76:M486–M492

    Article  CAS  PubMed  Google Scholar 

  • Nagar V, Shashidhar R, Bandekar JR (2013) Characterization of Aeromonas strains isolated from Indian foods using rpo D gene sequencing and whole cell protein analysis. World J Microbiol Biotechnol 29:745–752

    Article  CAS  PubMed  Google Scholar 

  • Nawaz M et al (2010) Detection and characterization of virulence genes and integrons in Aeromonas veronii isolated from catfish. Food Microbiol 27:327–331

    Article  CAS  PubMed  Google Scholar 

  • Pandove G, Sahota P, Vikal Y, Kaur B (2013) Multiplex PCR water testing kit for rapid, economic and simultaneous detection of Escherichia coli, Yersinia enterocolitica and Aeromonas hydrophila from drinking water. Curr Sci 104:352–358

    CAS  Google Scholar 

  • Pethiyagoda R (1991) Freshwater fishes of Sri Lanka: Wildlife Heritage Trust of Sri Lanka. Colombo, Sri Lanka

  • Popoff M, Veron MA (1976) A taxonomic study of the Aeromonas hydrophila-Aeromonas punctata group. Microbiol 94:11–22

    CAS  Google Scholar 

  • Rael RM, Frankenberger WT Jr (1996) Influence of pH, salinity, and selenium on the growth of Aeromonas veronii in evaporation agricultural drainage water. Water Res 30:422–430

    Article  CAS  Google Scholar 

  • Rahman M, Colque-Navarro P, Kühn I, Huys G, Swings J, Möllby R (2002) Identification and characterization of pathogenic Aeromonas veronii biovar sobria associated with epizootic ulcerative syndrome in fish in Bangladesh. Appl Environ Microbiol 68:650–655

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Raj NS et al (2019) Aeromonas veronii caused bilateral exophthalmia and mass mortality in cultured Nile tilapia, Oreochromis niloticus (L.) in India. Aquaculture 512:734278

    Article  Google Scholar 

  • Reavill D, Roberts H (2007) Diagnostic cytology of fish. Vet Clin Exot Ani Pract 10:207–234

    Article  Google Scholar 

  • Reed LJ, Muench HA (1938) A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27:493–497

    Article  Google Scholar 

  • Saad SM, Salem AM, Mahdy OA, Ibrahim ES (2019) Prevalence of metacercarial infection in some marketed fish in Giza Governorate, Egypt. J Egypt Soc Parasitol 49:129–134

    Article  Google Scholar 

  • Sahoo PK, Mohanty J, Das PC, Mohanta KN, Barik NK, Jayasankar P (2012) Fish health management. In: Sahoo PK et al (eds) CIFA: 25 years of freshwater aquaculture research. CIFA Bhubaneshwar, India, pp 147–178

    Google Scholar 

  • Samayanpaulraj V, Velu V, Uthandakalaipandiyan R (2019) Determination of lethal dose of Aeromonas hydrophila Ah17 strain in snake head fish Channa striata. Microb Pathog 127:7–11

    Article  CAS  PubMed  Google Scholar 

  • Sen K, Rodgers M (2004) Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities: a PCR identification. J Appl Microbiol 97:1077–1086

    Article  CAS  PubMed  Google Scholar 

  • Seshadri R et al (2006) Genome sequence of Aeromonas hydrophila ATCC 7966T: jack of all trades. J Bacteriol 188:8272–8282

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sha J, Kozlova EV, Chopra AK (2002) Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity. Infect Immun 70:1924–1935

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shameena SS, Kumar K, Kumar S, Kumar S, Rathore G (2020) Virulence characteristics of Aeromonas veronii biovars isolated from infected freshwater goldfish (Carassius auratus). Aquaculture 518:34819

    Article  Google Scholar 

  • Sreedharan K, Philip R, Singh IB (2011) Isolation and characterization of virulent Aeromonas veronii from ascitic fluid of Oscar Astronotus ocellatus showing signs of infectious dropsy. Dis Aquat Org 94:29–39

    Article  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Van Belkum A, Struelens M, de Visser A, Verbrugh H, Tibayrenc M (2001) Role of genomic typing in taxonomy, evolutionary genetics, and microbial epidemiology. Clin Microbiol Rev 14:547–560

    Article  PubMed  PubMed Central  Google Scholar 

  • Walker L, LeVine H, Jucker M (2006) Koch’s postulates and infectious proteins. Acta Neuropathol 112:1–4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang G, Tyler KD, Munro CK, Johnson WM (1996) Characterization of cytotoxic, hemolytic Aeromonas caviae clinical isolates and their identification by determining presence of a unique hemolysin gene. J Clin Microbiol 34:3203–3205

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang G et al (2003) Detection and characterization of the hemolysin genes in Aeromonas hydrophila and Aeromonas sobria by multiplex PCR. J Clin Microbiol 41:1048–1054

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wayne PA (2010) Clinical and Laboratory Standards Institute: Performance standards for antimicrobial susceptibility testing: 20th informational supplement (CLSI document M100-S20)

  • Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xia L et al (2008) Recovery of Acinetobacter baumannii from diseased channel catfish (Ictalurus punctatus) in China. Aquaculture 284:285–288

    Article  Google Scholar 

  • Xu XJ (1998) Role of a cytotoxic enterotoxin in Aeromonas-mediated infections: development of transposon and isogenic mutants. Infect Immun 66:3501–3509

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhu M et al (2016) Identification and virulence properties of Aeromonas veronii bv. sobria isolates causing an ulcerative syndrome of loach Misgurnusanguilli caudatus. J Fish Dis 39:777–781

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors acknowledge the guidance and support forwarded by Dr. Kishore Sarma, Research Associate, Department of Zoology, Gauhati University.

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  1. Department of Zoology, Gauhati University, Guwahati, Assam, 781014, India

    Rajdeep Das, Kangkan Sarma, Gayatree Hazarika, Hrishikesh Choudhury & Dandadhar Sarma

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RD and DS conceptualised and designed the experiments. RD, KS and GH conducted the experiments. RD, HC and DS analysed the results and drafted the manuscript. All authors read and reviewed the final manuscript.

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Correspondence to Dandadhar Sarma.

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Das, R., Sarma, K., Hazarika, G. et al. Identification and characterisation of emerging fish pathogens Aeromonas veronii and Aeromonas hydrophila isolated from naturally infected Channa punctata. Antonie van Leeuwenhoek 117, 4 (2024). https://doi.org/10.1007/s10482-023-01896-z

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