Investigation with Real-Time PCR and Histopathology on the presence of H. felis, H. heilmannii and H. pylori in dogs

  • Gokhan Akcakavak Yozgat Bozok University, Faculty of Veterinary Medicine, Department of Pathology Sorgun, Yosgat, Turkey
  • Mehmet Tuzcu Selcuk University, Faculty of Veterinary Medicine, Department of Pathology. Selcuklu, Konya, Turkey
  • Nevin Tuzcu Selcuk University, Faculty of Pharmacy, Department of Pharmaceutical Microbiology. Selcuklu, Konya, Turkey
  • Zeynep Celik Selcuk University, Faculty of Veterinary Medicine, Department of Pathology. Selcuklu, Konya, Turkey
  • Aysenur Tural Selcuk University, Faculty of Veterinary Medicine, Department of Pathology. Selcuklu, Konya, Turkey
  • Osman Dagar Selcuk University, Faculty of Veterinary Medicine, Department of Pathology. Selcuklu, Konya, Turkey
Keywords: Canine, histopathology, real-time PCR, H. felis, H. heilmannii, H. pylori

Abstract

Helicobacter species such as H. heilmannii, H. pylori, H. felis, H. bizzozeronii and H. salomonis have been identified in cats and dogs, and research suggesting that these species may be zoonotic agents and has been studied intensified in recent years. The aim of this study was to reveal the presence, comparing the histopathological findings and Real-time PCR results of H. felis, H. heilmannii, and H. pylori in the stomach and liver tissues taken during the necropsies of owned, stray or shelter dogs. The material of the study consisted of stomach and liver tissues taken from 35 dogs that died for different reasons and were brought for necropsy. DNA copies of H. heilmannii were detected by Real-time PCR in the liver samples of 30 dogs using H. heilmannii-specific primers. In the case of gastric samples, Real-time PCR detected H. heilmannii in 13 cases, H. pylori in 3 cases, both H. heilmannii and H. pylori in 13 cases, and H. felis, H. heilmannii and H. pylori in 3 cases. Microscopically, neutrophil leukocyte infiltration, epithelial degeneration, fibrosis and oedema in the lamina propia, and lymphoplasmacytic cell infiltration were determined in the stomachs. In the Hemotoxylin Eosin staining of the sections, 5 cases and 14 cases in the Warthin–Starry staining were found positive for Helicobacter-like microorganisms. Microscopically, dissociation of the remark cords and hydropic degeneration in hepatocytes, and focal mononuclear cell infiltrations in some sections were determined in the livers. In conclusion, with this study, it was understood that Real-time PCR analyzes are very useful in the diagnosis of H. felis, H. heilmannii, and H. pylori. However, histopathological examinations are necessary to associate the presence of bacteria with the development of the disease.

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References

AGUILAR, E.; ALVAREZ, M.; VIDAURRE, R.; RIVERA, M.; LEPE-LOPEZ, M.; Trends in the veterinary clinic for pets in guatemala city. Rev. Cientif. FCV-LUZ. XXX(4): 186–191. 2020.

ARMSTRONG, D. Helicobacter pylori infection and dyspepsia. Scand. J. Gastroenterol. Suppl. 31: 38–47. 1996.

ASHAOLU, J.O.; TSAI, Y.J.; LIU, C.C.; JI, D.D. Prevalence, diversity and public health implications of Helicobacter species in pet and stray dogs. One Health. 15: 100430. 2022. https://doi.org/jvnh.

BRUYETTE, D. Clinical Small Animal Internal Medicine, In: Gastritis and Gastric Ulceration in dogs and cats, John Wiley & Sons. Pp. 547–555. 2020.

BURUCOA, C.; AXON, A. Epidemiology of Helicobacter pylori infection. Helicob. 22: e12403. 2017. https://doi.org/gg6dq6.

DİKER, K.S.; HAZIROĞLU, R.; AKAN, M.; ÇELİK, S.; KABAKÇI, N. The prevalence, colonization sites and pathological effects of gastric helicobacters in dogs. Turkish J. Vet. Anim. Sci. 26: 345–351. 2002.

DUNN, B.E. Pathogenic mechanisms of Helicobacter pylori. Gastroenterol. Clin. North Am. 22: 43–57. 1993.

EATON, K.; DEWHIRST, F.; PASTER, B.; TZELLAS, N.; COLEMAN, B.; PAOLA, J.; SHERDING, R. Prevalence and varieties of Helicobacter species in dogs from random sources and pet dogs: animal and public health implications. J. Clin. Microbiol. 34: 3165–3170. 1996.

GERMANI, Y.; DAUGA, C.; DUVAL, P.; HUERRE, M.; LEVY, M.; PIALOUX, G.; SANSONETTI, P.; GRIMONT, P. Strategy for the detection of Helicobacter species by amplification of 16S rRNA genes and identification of H. felis in a human gastric biopsy. Res. Microbiol. 148: 315–326. 1997.

HAESEBROUCK, F.; PASMANS, F.; FLAHOU, B.; CHIERS, K.; BAELE, M.; MEYNS, T.; DECOSTERE, A.; DUCATELLE, R. Gastric helicobacters in domestic animals and nonhuman primates and their significance for human health. Clin. Microbiol. Rev. 22: 202–223. 2009.

HÄNNINEN, M.L.; HAPPONEN, I.; SAARI, S.; JALAVA, K. Culture and characteristics of Helicobacter bizzozeronii, a new canine gastric Helicobacter spp.. Int. J. Syst. Evol. Microbiol. 46: 160–166. 1996.

HAPPONEN, I.; LINDEN, J.; SAARI, S.; KARJALAINEN, M.; HÄNNINEN, M.L.; JALAVA, K.; WESTERMARCK, E. Detection and effects of helicobacters in healthy dogs and dogs with signs of gastritis. J. Am. Vet. Med. Assoc. 213: 1767–1774. 1998.

HAPPONEN, I.; SAARI, S.; CASTREN, L.; TYNI, O.; HÄNNINEN, M.L.; WESTERMARCK, E. Occurrence and topographical mapping of gastric Helicobacter‐like organisms and their association with histological changes in apparently healthy dogs and cats. J. Vet. Med. 43: 305–315. 1996.

HERMANNS, W.; KREGEL, K.; BREUER, W.; LECHNER, J. Helicobacter-like organisms: histopathological examination of gastric biopsies from dogs and cats. J. Comp. Pathol. 112: 307–318. 1995.

HLAOPERM, C.; CHOOWONGKOMON, K.; PRUKSAKORN, C.; RATTANASRISOMPORN, J. Development of an easy-to-use urease kit for detecting Helicobacter pylori in canine gastric mucosa. Vet. World. 14: 1977. 2021.

HUSNIK, R.; KLIMES, J.; KOVARIKOVA, S.; KOLORZ, M. Helicobacter Species and Their Association with Gastric Pathology in a Cohort of Dogs with Chronic Gastrointestinal Signs. Anim. 12: 1254. 2022.

HWANG, C.Y.; HAN, H.R.; YOUN, H.Y. Prevalence and clinical characterization of gastric Helicobacter species infection of dogs and cats in Korea. J. Vet. Sci. 3: 123–133. 2002.

JALAVA, K.; KAARTINEN, M.; UTRIAINEN, M.; HAPPONEN, I.; HÄNNINEN, M.L. Helicobacter salomonis spp.. nov., a canine gastric Helicobacter spp., related to Helicobacter felis and Helicobacter bizzozeronii. Int. J. Syst. Evol. Microbiol. 47: 975–982. 1997.

JALAVA, K.; ON, S.L.; VANDAMME, P.A.; HAPPONEN, I.; SUKURA, A.; HÄNNINEN, M.L. Isolation and identification of Helicobacter spp. from canine and feline gastric mucosa. Appl. Environ. Microbiol. 64: 3998–4006. 1998.

KAUSER, F.; HUSSAIN, M.A.; AHMED, I.; SRINIVAS, S.; DEVI, S.M.; MAJEED, A.A.; RAO, K.R.; KHAN, A.A.; SECHI, L.A.; AHMED, N. Comparative genomics of Helicobacter pylori isolates recovered from ulcer disease patients in England. BMC Microbiol. 5: 1–10. 2005.

LADEIRA, M.; SALVADORI, D.; RODRIGUES, M. Biopathology of Helicobacter pylori. J. Bras. Patol. Med. Lab. 39: 335–342. 2003. https://doi.org/dp78t6.

LUNA, L. Manual of histologic staining methods of the Armed Forces Institute of Pathology, In: Routine Staining Procedures. Blakiston Division, McGraw-Hill, New York. 258 pp. 1968.

MURRAY, P.R.; BARON, E.J.; PFALLER, M.A.; TENOVER, F.C.; YOLKEN, R.H.; MORGAN, D.R. Vibrio and realted organism. Manual of Clinical Microbiology. 6rd. Ed. 1482 pp. 1995.

NEIGER, R.; DIETERICH, C.; BURNENS, A.; WALDVOGEL, A.; CORTHÉSY-THEULAZ, I.; HALTER, F.; LAUTERBURG, B.; SCHMASSMANN, A. Detection and prevalence of Helicobacter infection in pet cats. J. Clin. Microbiol. 36: 634–637. 1998.

NEIGER, R.; SIMPSON, K.W. Helicobacter infection in dogs and cats: facts and fiction. J. Vet. Intern. Med. 14: 125–133. 2000.

NEIGER, R.; TSCHUDI, M.E.; BURNENS, A.; GOKE, B.; SCHMASSMANN, A. Diagnosis and identification of gastric Helicobacter species by polymerase chain reaction in dogs. Microb. Ecol. Health Dis. 11: 234–240. 1999.

NORRIS, C.; MARKS, S.L.; EATON, K.; TORABIAN, S.; MUNN, R.; SOLNICK, J.V. Healthy cats are commonly colonized with “Helicobacter heilmannii” that is associated with minimal gastritis. J. Clin. Microbiol. 37: 189–194. 1999.

OKUBO, B.M.; RICCI-AZEVEDO, R.; ZOBIOLE, N.N.; BUCCINI, D.F.; MORENO, S.E. Prevalência de Helicobacter spp. em cães de Campo Grande-MS. Ciên. Anim. Bras. 18: e-17286. 2017. https://doi.org/jvnj.

ONUR, E. İstanbul’da kedi köpek sahiplenmesinin sosyo-ekonomik parametreleri. İstanbul Üniversitesi Sağlık Bilimleri Enstitüsü, Turkey. Doctoral Thesis. 100 pp. 2012.

OTTO, G.; HAZELL, S.; FOX, J.; HOWLETT, C, MURPHY.; J, O'ROURKE.; J, LEE A. Animal and public health implications of gastric colonization of cats by Helicobacter-like organisms. J. Clin. Microbiol. 32: 1043–1049. 1994.

ÖZEN, H.; KARAMAN, M.; ÖZCAN, K. Detection and prevalence of gastric Helicobacters in dogs: comparasion of may-Grünwald Giemsa, warthin-starry and immunohistochemistry. Vet. 16: 48–56. 2005.

SAĞNAK, S.; ÖZGÜR, N.Y. İstanbul Yöresinde Sağlıklı ve Klinik Belirti Gösteren Köpeklerde Helicobacter pylori Varlığının Kültür, Polimerase Chain Reaction (PCR) ve Restriction Fragment Length Polymorphism (RFLP) ile Saptanması. İstanbul Üniversitesi Sağlik Bilimleri Enstitüsü. Turkey. Doctoral Thesis. 56 pp. 2011.

TABRIZI, A.S.; DERAKHSHANDEH, A.; ESFANDIARI, A.; ATASHI Z.A. Identification of Helicobacter spp. in gastrointestinal tract, pancreas and hepatobiliary system of stray cats. Iran. J. Vet. Res. 16: 374. 2015.

XIA, H.X.; KEANE, C.T.; CHEN, J.; ZHANG, J.; WALSH, E.J.; MORAN, A.P.; HUA, J.S.; MEGRAUD, F.; O'MORAIN C.A. Transportation of Helicobacter pylori cultures by optimal systems. J. Clin. Microbiol. 32: 3075–3077. 1994.

YAMASAKI, K.; SUEMATSU, H.; TAKAHASHI, T. Comparison of gastric lesions in dogs and cats with and without gastric spiral organisms. J. Am. Vet. Med. Assoc. 212: 529–533. 199.

Real time PCR Plot
Published
2023-02-07
How to Cite
1.
Akcakavak G, Tuzcu M, Tuzcu N, Celik Z, Tural A, Dagar O. Investigation with Real-Time PCR and Histopathology on the presence of H. felis, H. heilmannii and H. pylori in dogs. Rev. Cient. FCV-LUZ [Internet]. 2023Feb.7 [cited 2025Aug.2];33(1):1-. Available from: http://www.produccioncientifica.luz.edu.ve/index.php/cientifica/article/view/39656
Section
Veterinary Medicine