Skip to main content
SpringerLink
Log in

Isolation and characterization of Mycoplasmas from some moribund Egyptian fishes

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

Mycoplasma is associated with fatal diseases in human, animals, and fish. Mycoplasma of aquatic origin has not been thoroughly studied. The current study has shed the light on isolation and characterization of unique piscine Mycoplasma isolates retrieved from diverse geographical locations across the Egyptian state. Mycoplasma spp. was isolated using specific culture media. Mycoplasma was identified using the morphochemical tests and then confirmed molecularly by PCR of the 16S rRNA gene. The results indicated that the incidence of Mycoplasma from Cyprinus carpio, Oreochromis niloticus, Aulopiformes synodontida, and Clarias gariepinus were 33.33%, 16.36%, 8.108%, and 6.45%, respectively, while Mugil cephalus were negative for Mycoplasma isolation. Mycoplasma was detected only from gills and swim bladder of affected fish. Biochemically, the isolated Mycoplasmas were grouped into two clusters. Mycoplasma cluster 1 (35 isolates) and Mycolplasma cluster 2 (7 isolates). Mycoplasma cluster 1 was positive for tetrazolium reduction while Mycoplasma cluster 2 was negative. The phylogenetic tree of partial sequences of 16S rRNA showed that both clusters were grouped in one branch and separated from other Mycoplasma spp., suggesting that both clusters are belonging to one species. Interestingly, all fish Mycoplasma isolates were PCR negative for both Mycoplasma mobile and Mycoplasma monodon using specific species primers. This result confirmed that these two clusters belonged to unspecified Mycoplasma species, for which the temporary names Mycoplasma cluster 1 and Mycoplasma cluster 2 were designated. Pathogenicity trials of both Mycoplasmas clusters revealed that all inoculated Nile tilapias were susceptible to the unspecified Mycoplasmas.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Abbas AA, El-Shabiny LM, Shaheen AA (2000) Studies on mycoplasmosis among some fishes. Suez Canal Vet Med J 3(2):705–716

    Google Scholar 

  • Ahmed HA, El-Hady, Maha A. (2008) Antigenic variations and pathogenicity between Mycoplasma spp. isolated from fishes and shellfishes. 8th International Symposium on Tilapia Aquaculture.Cairo International Convention Center (CICC). Egypt 12–14 October Volume (2): 1229–1243. (ISTA VIII)

  • Austin B, Austin DA (2007) Bacterial fish pathogens, diseases of farmed and wild fish, 4th edn. Praxis Publishing Ltd, Chichester

    Google Scholar 

  • Bano N, Smith AR, Bennett W et al (2007) Dominance of Mycoplasma in the guts of the long-jawed mudsucker, Gillichthys mirabilis, from five California salt marshes. Environ Microbiol 9:2636–2641

    CAS  PubMed  Google Scholar 

  • Baseman JB, Tully JG (1997) Mycoplasmas: sophisticated, reemerging and burdened by their notoriety. Emerg Infect Dis 3:21–32

    CAS  PubMed  PubMed Central  Google Scholar 

  • Black FT (1973) Phosphatase activity in T-mycoplasmas. Int J Syst Bacteriol 23(1):65–66

    Google Scholar 

  • Blanchard A, Gautier M, Mayau V (1991) Detection and identification of mycoplasmas by amplification of rDNA. FEMS Microbiol Lett 65:37–42

    CAS  PubMed  Google Scholar 

  • Brown DR (2002) Mycoplasmosis and immunity of fish and reptiles. Front Biosci 1(7):d 1338–d 1346

    Google Scholar 

  • Brown RM, Wiens GD, Salinas I (2019) Analysis of the gut and gill microbiome of resistant and susceptible lines of rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immunol 86:497–506. https://doi.org/10.1016/j.fsi.2018.11.079

    Article  CAS  PubMed  Google Scholar 

  • Cole BC, Ward JR, Martin CH (1968) Hemolysin and peroxide activity of Mycoplasma species. J Bacteriol 95:2022–2030

    CAS  PubMed  PubMed Central  Google Scholar 

  • Conrads G, Flemmig TF, Seyfarth I et al (1999) Simultaneous detection of Bacteroidesforsythus and Prevotellaintermedia by 16S rRNA gene-directed multiplex PCR. J Clin Microbiol 37:1621–1624

    CAS  PubMed  PubMed Central  Google Scholar 

  • Deng S, Hiruki C, Robertson JA, Stemke GW (1992) Detection by PCR and differentiation by restriction fragment length polymorphism of Acholeplasma, Spiroplasma, Mycoplasma, and Ureaplasma, based upon 16S rRNA genes. PCR Methods Appl 1:202–204

    CAS  PubMed  Google Scholar 

  • Doyle S, Macdonald B, Rochette R (2010) Is water temperature responsible for geographic variation in shell mass of Littorinaobtusata (L.) snails in the Gulf of Maine? J Exp Mar Biol Ecol 394:98–104

    Google Scholar 

  • Eissa AE (2016) Clinical and Laboratory Manual of Fish Diseases. LAP LAMBERT Academic Publishing

  • El-Shabiny LM, Ibrahim MM, Mahmoud NA (1989) Mycoplasma as fish pathogen in Egypt. Vet Med J Giza 37(3):335–343

    Google Scholar 

  • El-Shabiny LM, Mahmoud NAM, Alyain SA, El-Sharnouby R (1996) Mycoplasma as a causative pathogenic agent in Clarias lazera in Egypt. Vet Med J Giza 44(4):657–662

    Google Scholar 

  • Erno H, Stipkovits L (1973) Bovine Mycoplasma cultural and biochemical studies. Acta Vet Scand 14:450–463

    CAS  PubMed  Google Scholar 

  • Fabricant J, Freundt EA (1967) Importance of extension and standardization of laboratory tests for the identification and classification of Mycoplasma. Ann N Y Acad Sci 143:50–58

    CAS  PubMed  Google Scholar 

  • Fan HH, Kleven SH. And Jackwood MW.1995. Application of polymerase chain reaction with arbitrary primers to strain identification of Mycoplasma gallisepticum. Avian Dis 39:729–735

    CAS  PubMed  Google Scholar 

  • Freundt EA (1973) Principles of Mycoplasma classification. Ann N Y Acad Sci 225:713

    Google Scholar 

  • Frey ML, Hanson RP, Anderson DP (1968) A medium for isolation of avian Mycoplasmas. Am J Vet Res 29:2163–2171

    CAS  PubMed  Google Scholar 

  • Geary SJ, Forsyth MH, Abul Soaud S et al (1994) Mycoplasmagallisepticum strains differentiation by arbitrary primer PCR (RAPD) fingerprinting. Mol Cell Probes 8:311–316

    CAS  PubMed  Google Scholar 

  • Ghadersohi A, Owens L (1999) Isolation, characterisation and DNA analysis of Mycoplasma spp. from moribund prawns Penaeusmonodon cultured in Australia. Dis Aquat Org 35(1):53–61

    CAS  Google Scholar 

  • Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp 41:95–99

    CAS  Google Scholar 

  • Hata DJ (2010) Molecular methods for identification and characterization of Acinetobacter spp. In: Molecular Diagnostics. p 313–326

  • Hosseini H, Bozorgmehrifard MH, Peighambari SM et al (2006) Random amplified polymorphic DNA (RAPD) fingerprinting of Mycoplasma gallisepticum isolates from chickens. Arch Razi Inst 61:67–71

    CAS  Google Scholar 

  • Husien MM, Shaker MM, Marzouk MS (1998) Preliminary studies on mycoplasma infection in Nile carp (Labeo niloticus) in Egypt. Vet Med J Giza 46(4.A):407–416

    Google Scholar 

  • Jaffe JD, Stange-Thomann N, Smith C (2004) The complete genome and proteome of Mycoplasma mobile. Genome Res 14(8):1447–1461

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kaewjam RS (1987) The apple snails of Thailand: aspects of comparative anatomy. Malacol Rev 20:69–89

    Google Scholar 

  • Kashinskaya EN, Simonov EP, Izvekova GI, Parshukov AN, Andree KB, Solovyev MM (2019) Composition of the microbial communities in the gastrointestinal tract of perch (Perca fluviatilis L. 1758) and cestodes parasitizing the perch digestive tract. J Fish Dis 00:1–16. https://doi.org/10.1111/jfd.13096

    Article  CAS  Google Scholar 

  • Kim DH, Brunt J, Austin B (2007) Microbial diversity of intestinal contents and mucus in rainbow trout (Oncorhynchus mykiss). J Appl Microbiol 102:1654–1664

    CAS  PubMed  Google Scholar 

  • Kusumoto A, Seto S, Jaffe JD, Miyat M (2004) Cell surface differentiation of Mycoplasma mobile visualized by surface protein localization. Microbiol. 150(Pt12):4001–4008

    CAS  Google Scholar 

  • Ley DH, Berkhoff JE, Levisohn S (1997) Molecular epidemiological investigations of Mycoplasmagallisepticum conjunctivitis in songbirds by random amplified polymorphic DNA analysis. Emerg Infect Dis 3:375–380

    CAS  PubMed  PubMed Central  Google Scholar 

  • Li X, Yan Q, EinarRingø E et al (2016) The influence of weight and gender on intestinal bacterial community of wild largemouth bronze gudgeon (Coreiusguichenoti, 1874). BMC Microbiol 16:191

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu WH, Chiu YW, Huang DJ, Liu MY, Lee CC, Liu LL (2006) Imposex in the golden apple snail Pomacea canaliculata in Taiwan. Sci Total Environ 371:138–143

    CAS  PubMed  Google Scholar 

  • Moran D, Turner SJ, Clements KD (2005) Ontogenetic development of the gastrointestinal microbiota in the marine herbivorous fish Kyphosus sydneyanus. Microb Ecol 49:590–597

    CAS  PubMed  Google Scholar 

  • Ohtani N, Miyata M (2007) Identification of a novel nucleoside triphosphatase from Mycoplasma mobile: a prime candidate motor for gliding motility. Biochem J 403(1):71–77

    CAS  PubMed  PubMed Central  Google Scholar 

  • Razin S (2006) Chapter 1.2.29.The genus mycoplasma and related genera (class Mollicutes), The Prokaryotes. https://doi.org/10.1007/0-387-30744-3

  • Razin S, Yogev D, Naot Y (1998) Molecular biology and pathogenicity of mycoplasmas. Microbiol Mol Biol Rev 62:1094–1156

    CAS  PubMed  PubMed Central  Google Scholar 

  • Rollinson D, Stothard JR, Jones CS et al (1998) Molecular characterisation of intermediate snail hosts and the search for resistance genes. Mem Inst Oswaldo Cruz 93:111–116

    PubMed  Google Scholar 

  • Sabry M (1968) Characterization of avian Mycoplasma. Unpublished Ph.D. thesis. Cornell University, Ithaca, New York

  • Sabry MZ, Erno H, Freundt EA (1971) Manual of technical methods for the characterization and serotyping of Mycoplasma.Ed. by Mycoplasma Division, Ani. Health Res. Inst. of Agric., Dokki, Giza, Egypt

  • Sasaki Y (2006) Mycoplasma. In: Chan VL, Sherman PM, Bourke B (eds) Bacterial Genomes and Infectious Diseases. Humana Press Inc., Totowa

    Google Scholar 

  • Sievers F, Wilm A, Dineen D et al (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7:539

    PubMed  PubMed Central  Google Scholar 

  • Subhraveti P, QuangOng Q, Keseler I et al. (2017) Summary of Mycoplasma mobile, strain 163K, version 21.5. PGDB unique ID: 1G0Z

  • Sukhanova EV, Dzyuba EV, Triboy TI et al (2011) Molecular genetic and cultural diagnostics of Mycoplasma in fishes of the family Thymallidae. Dokl Biol Sci 440:270–272

    Google Scholar 

  • Sukhanova EV, Denikina NN, Triboy TI et al (2014) Molecular and phylogenetic studies of a Mycoplasma from the intestine of Siberian fish. Bio Genet J 2(1):37–41

    Google Scholar 

  • Timenetsky J, Santos LM, Buzinhani M, Mettifogo E (2006) Detection of multiple Mycoplasma infection in cell cultures by PCR. Braz J Med Biol Res 39:907–914

    CAS  PubMed  Google Scholar 

  • Uenoyama A, Kusumoto A, Miyata M (2004) Identification of a 349-kilodalton protein (Gli349) responsible for cytadherence and glass binding during gliding of Mycoplasma mobile. Bacteriol. 186(5):1537–1545

    CAS  Google Scholar 

  • Volokhov DV, George J, Liu SX, Ikonomi P, Anderson C, Chizhikov V (2006) Sequencing of the intergenic 16S–23S rRNA spacer (ITS) region of Mollicutes species and their identification using microarray-based assay and DNA sequencing. Appl Microbiol Biotechnol 71:680–698

    CAS  PubMed  Google Scholar 

  • Wang W, Gu W, Ding Z et al (2005) A novel Spiroplasma pathogen causing systemic infection in the crayfish Procambarusclarkii (Crustacea: Decapod), in China. FEMS Microbiol Lett 249:131

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology, Faculty of Veterinary Medicine, Cairo University, Giza, 11221, Egypt

    Jakeen EI-Jakee & Sarah Elshamy

  2. Mycoplasma Department, Animal Health Research Institute, Dokki, Giza, Egypt

    Abdel-Wahed Hassan

  3. Department of Fish Diseases and Management, Faculty of Veterinary Medicine, Cairo University, Giza, 11221, Egypt

    Mohamed Abdelsalam, Nehal Younis & Alaa Eldin Eissa

  4. Fish Diseases Department, Animal Health Research Institute, Dokki, Giza, Egypt

    Maha A. El-Hady

Authors
  1. Jakeen EI-Jakee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sarah Elshamy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Abdel-Wahed Hassan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mohamed Abdelsalam
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nehal Younis
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Maha A. El-Hady
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Alaa Eldin Eissa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alaa Eldin Eissa.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical considerations

All standard national and international ethical guidelines dealing with fish handling, challenge, and sampling were fully adopted by the research team (authors).

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(DOCX 18 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

EI-Jakee, J., Elshamy, S., Hassan, AW. et al. Isolation and characterization of Mycoplasmas from some moribund Egyptian fishes. Aquacult Int 28, 901–912 (2020). https://doi.org/10.1007/s10499-019-00502-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-019-00502-2

Keywords

Search

Navigation