Vet Med - Czech, 2023, 68(2):69-74 | DOI: 10.17221/85/2022-VETMED

Verification of accuracy of qPCR method for intravital diagnostics of Macrorhabdus ornithogaster in avian droppingsOriginal Paper

L Vrbasova1, D Molinkova2, P Linhart3, Z Knotek1
1 Avian and Exotic Animal Clinic
2 Department of Infectious Diseases and Microbiology
3 Department of Animal Protection and Welfare and Veterinary Public Health, VETUNI Brno, Czech Republic

The objective of this study was to verify the accuracy of the quantitative PCR (qPCR) method for in vivo detection of Macrorhabdus ornithogaster. A total of 100 faecal and gastric mucosa samples from avian cadavers were investigated, using cytological and qPCR techniques (budgerigars, Fischer’s lovebirds, red-crowned parakeets, scarlet-chested parrots, eastern rosellas, domestic canaries, zebra finches, white Java sparrow). Using qPCR, the probability of detecting positive samples of droppings was significantly higher than in the faecal smear microscopy (P < 0.01). Cytology detected the presence of Macrorhabdus ornithogaster in 34 faecal samples, whereas qPCR detected 54 positive samples. In all 46 qPCR negative faecal samples, gastric smear qPCR was performed and also yielded negative results. Gastric smear qPCR was also performed in 20 cadavers where faecal qPCR has detected the presence of Macrorhabdus ornithogaster and in all samples confirmed the positive result. This verifies the accuracy of faecal sample qPCR for intravital diagnostics. Overall, the faecal qPCR technique appears to be extremely reliable, as it made it possible to detect all infected individuals, including those with negative stool or gastric cytology.

Keywords: cytology; macrorhabdosis; passerines; psittacines

Received: October 21, 2022; Accepted: January 27, 2023; Prepublished online: February 22, 2023; Published: February 24, 2023  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Vrbasova L, Molinkova D, Linhart P, Knotek Z. Verification of accuracy of qPCR method for intravital diagnostics of Macrorhabdus ornithogaster in avian droppings. Vet Med - Czech. 2023;68(2):69-74. doi: 10.17221/85/2022-VETMED.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Baron HR, Stevenson BC, Phalen DN. Inconsistent efficacy of water-soluble amphotericin B for the treatment of Macrorhabdus ornithogaster in a budgerigar (Melopsittacus undulatus) aviary. Aust Vet J. 2020 Jul;98(7):333-7. Go to original source... Go to PubMed...
    2. Borrelli L, Dipineto L, Rinaldi L, Romano V, Noviello E, Menna LF, Cringoli G, Fioretti A. New diagnostic insights for Macrorhabdus ornithogaster infection. J Clin Microbiol. 2015 Nov;53(11):3448-50. Go to original source... Go to PubMed...
    3. Filippich LJ, Hendrikz JK. Prevalence of megabacteria in budgerigar colonies. Aust Vet J. 1998 Feb;76(2):92-5. Go to original source... Go to PubMed...
    4. Gerlach H. Megabacteriosis. Semin Avian Exot Pet Med. 2001 Jan;10(1):12-9. Go to original source...
    5. Hanafusa Y, Bradley A, Tomaszewski EE, Libal MC, Phalen DN. Growth and metabolic characterization of Macrorhabdus ornithogaster. J Vet Diagn Invest. 2007 May;19(3):256-65. Go to original source... Go to PubMed...
    6. Hanafusa Y, Costa E, Phalen DN. Infection trials in mice suggest that Macrorhabdus ornithogaster is not capable of growth in mammals. Med Mycol. 2013 Aug;51(6):669-72. Go to original source... Go to PubMed...
    7. Hanka K, Kohler K, Kaleta EF, Sommer D, Burkardt E. Macrorhabdus ornithogaster: Nachweise bei Ziervogeln, Hausgeflugel und Stadttauben sowie morphologische Charakterisierung und Versuche zur in-vitro-Kultivierung [Macrorhabdus ornithogaster: Detection in companion birds, poultry and pigeons, morphological characterisation and examination of in vitro cultivation]. Prakt Tierarzt. 2010;91(5):390-400. German.
    8. Kheirandish R, Salehi M. Megabacteriosis in budgerigars: Diagnosis and treatment. Comp Clin Pathol. 2011;20(5):501-5. Go to original source...
    9. Lanzarot P, Blanco JL, Alvarez-Perez S, Abad C, Cutuli MT, Garcia ME. Prolonged fecal shedding of 'megabacteria' (Macrorhabdus ornithogaster) by clinically healthy canaries (Serinus canaria). Med Mycol. 2013 Nov;51(8):888-91. Go to original source... Go to PubMed...
    10. Marlier D, Leroy C, Sturbois M, Delleur V, Poulipoulis A, Vindevogel H. Increasing incidence of megabacteriosis in canaries (Serinus canarius domesticus). Vet J. 2006 Nov;172(3):549-52. Go to original source... Go to PubMed...
    11. Martins NRS, Horta AC, Siqueira AM, Lopes SQ, Resende JS, Jorge MA, Assis RA, Martins NF, Fernandes AA, Barrios PR, Costa TJR, Guimaraes LMC. Macrorhabdus ornithogaster in ostrich, rhea, canary, zebra finch, free range chicken, turkey, guinea-fowl, columbina pigeon, toucan, chuckar partridge and experimental infection in chicken, japanese quail and mice. Arq Bras Med Vet Zootec. 2006;58(3):291-8. Go to original source...
    12. Phalen DN, Moore RP. Experimental infection of white-leghorn cockerels with Macrorhabdos ornithogaster (Megabacterium). Avian Dis. 2003 Apr-Jun;47(2):254-60. Go to original source...
    13. Phalen DN. Update on the diagnosis and management of Macrorhabdus ornithogaster (formerly megabacteria) in avian patients. Vet Clin North Am Exot Anim Pract. 2014 May;17(2):203-10. Go to original source... Go to PubMed...
    14. Piesacki T, Prochwska S, Celmer Z, Sochacka A, Bednarski M. Wystepowanie Macrorhabdus ornithogaster u ptakow ozdobnych oraz dziko zyjacych w Polsce [Occurrence of Macrorhabdus ornithogaster in exotic and wild birds in Poland]. Med Weter. 2012;68(4):245-8. Polish.
    15. Poleschinski JM, Straub JU, Schmidt V. Comparison of two treatment modalities and PCR to assess treatment effectiveness in macrorhabdosis. J Avian Med Surg. 2019 Sep 9;33(3):245-50. Go to original source... Go to PubMed...
    16. Powers LV, Mitchell MA, Garner MM. Macrorhabdus ornithogaster infection and spontaneous proventricular adenocarcinoma in budgerigars (Melopsittacus undulatus). Vet Pathol. 2019 May;56(3):486-93. Go to original source... Go to PubMed...
    17. Pustow R, Krautwald-Junghanns ME. The incidence and treatment outcomes of Macrorhabdus ornithogaster infection in budgerigars (Melopsittacus undulatus) in a veterinary clinic. J Avian Med Surg. 2017 Dec;31(4):344-50. Go to original source... Go to PubMed...
    18. Sullivan PJ, Ramsay EC, Greenacre CB, Cushing AC, Zhu X, Jones MP. Comparison of two methods for determining prevalence of Macrorhabdus ornithogaster in a flock of captive budgerigars (Melopsittacus undulatus). J Avian Med Surg. 2017 Jun;31(2):128-31. Go to original source... Go to PubMed...
    19. Tomaszewski EK, Logan KS, Snowden KF, Kurtzman CP, Phalen DN. Phylogenetic analysis identifies the 'megabacterium' of birds as a novel anamorphic ascomycetous yeast, Macrorhabdus ornithogaster gen. nov., sp. nov. Int J Syst Evol Microbiol. 2003 Jul;53(Pt 4):1201-5. Go to original source... Go to PubMed...
    20. Vrbasova L, Molinkova D, Obdrzalek D, Grosova T, Knotek Z. Vyuziti metody real-time PCR pri diagnostice a kontrole ucinnosti lecby makrorabdozy [Use of real-time PCR method in the diagnostics and control of the effectiveness of macrorabdosis treatment]. Veterinární klinika. 2020;17(6):216-21. Czech.

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content