Ali ASL SHABESTARI, Hadi AMANIZAD, Alireza PARTOVI NEJAD, Mehdi JALILI, Dariush NEZHAD MOHAMMAD, Mohammad Hosein SOROUSH, Abolfazl BARZEGARI, Daryoush BABAZADEH

Assessment of gastric helicobacter spp. in fresh gastric samples of naturally infected dogs by scanning electron microscopy

Köpeklerin midesinde Gastrik Helicobacter-benzeri Organizmalar (GHBO)ın üç farklı türü olduğu bilinmektedir. Bu çalışmanın amacı, doğal enfekte köpeklerin taze mide içeriklerinde gastrik Helicobacter türlerinin morfolojik özelliklerinin belirlenmesidir. Sokak köpeklerinden nekropsi sırasında otuz iki adet mide içeriği örneği alındı. Örnekler Hızlı Üreaz Testi, Işık Mikroskopisi, Taramalı Elektron Mikroskopisi (SEM) ve Zincirleme Polimeraz Reaksiyonu (PCR) ile incelendi. Işık Mikroskopisi ile köpeklerin %90.5inde Gastrik Helicobacter- benzeri Organizmaların varlığı belirlendi. Köpeklerin %87.5i ve %94ü sırası ile Hızlı Üreaz Testi ve PCR pozitif bulundu. Dört adet Helicobacter bakterisi SEM ile tespit edildi. Bu bakterilerden 3 suş, PCR ve morfolojik farklılıklarına göre H. felis, candidatus H. heilmanii ve H. Bizzozeronii olarak tanımlandı. Bakterinin son suşu ise rutin metodlar ile identifiye edilemedi. Köpeklerin Gastrik Helicobacterlerinin tiplerini belirlemek için hızlı ve basit tanı metodları üzerinde yapılacak geniş çaplı çalışmalara ihtiyaç vardır. Bu çalışmanın sonuçları yeni Helicobacter türlerinin varlığının bildirilmesinden dolayı köpeklerin GHBOnın daha detaylı araştırılması gereğini ortaya koymaktadır.

Doğal enfekte köpeklerin taze mide içeriği örneklerinde gastrik helicobacter Türlerinin taramalı elektron mikroskopi ile değerlendirilmesi

Diferent species of gastric Helicobacter-Like Organisms (GHLO) reported from dogs stomach. The aim of present study was to morphological evaluation of gastric Helicobacter spp. in fresh gastric samples of naturally infected dogs. Thirty two gastric samples of the stray dog were taken at necropsy. The specimens were used for rapid urease test, light microscopy, scanning electron microscopy (SEM) and polymerase chain reaction (PCR). Light microscopy examination confirmed the presence of GHLO in 90.5% of stray dogs. 87.5% and 94% of gastric samples were positive in rapid urease test and PCR, respectively. Four distinguishable Helicobacter organisms were confirmed by SEM. Three strains of these organisms were inditified as H. felis, candidatus H. heilmanii and H. bizzozeronii because of their apparent morphological diferences and PCR results. The last strain of these bacteria was not distinguishable with routine studies. The large-scale studies with fast and simple recognition methods are recommended to confirm the diferent types of canine gastric Helicobacter. The results of present study showed further investigation in canine GHLO is required because new species of Helicobacter reported.

PDF

___

1. Lavelle JP, Landas S, Mitros FA, Conklin JL: Acute gastritis associated with spiral organisms from cats. Digest Dis Sci, 39, 744-750, 1994.
2. Eaton KA, Dewhirst FE, Paster BJ, Tzellas N, Coleman BE, 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.
3. Fox JG, Lee A: The role of Helicobacter species in newly recognized gastrointestinal tract diseases of animals. J Lab Anim Sci, 47, 222-255, 1997.
4. Jalava K, Kaartinen M, Utriainen M, Happonen I, Hanninen ML: Helicobacter salomonis sp. Nov., a canine gastric Helicobacter sp. related to Helicobacter felis, and Helicobacter bizzozeronii. Int J Syst Bacteriol, 47, 975-982, 1997.
5. Simpson KW, McDonough PL, Strauss-Ayali D, Chang YF, Harpending P, Valentine BA: Helicobacter felis infection in dogs: Efect on gastric structure and function. J Vet Pathol, 36, 237-248, 1999.
6. Prachasilpchai W, Nuanualsuwan S, Chatsuwan T, Techangamsuwan S, Wangnaitham S, Sailasuta A: Diagnosis of Helicobacter spp. infection in canine stomach. J Vet Sci, 8, 139-145, 2007.
7. Robic M, Artuković B, Beck A, Gudan A, Svetina A, Grabarević Z: Histopatological changes in stomachs of dogs with naturally acquired Helicobacter infection. Vet Arhiv, 77, 103-111, 2007.
8. Haesebrouck F, Pasmans F, Flahou B, Chiers K, Baele M, Meyns T, Decostere A, Ducatelle R: Gastric Helicobacters in domestic animals and nonhuman primates: The agents and their significance for human health. Clin Microbiol, 22, 202-223, 2009.
9. Seymour C, Lewis RG, Kim M, Gagnon DF, Fox JG, Dewhirst FE, Paster ANDBJ: Isolation of Helicobacter strains from wild bird and swine feces. Appl J Environ Microbiol, 60, 1025-1028, 1994.
10. Harper CMG, Dangler CA, Xu S, Feng Y, Shen Z, Sheppard B, Stamper A, Dewhirst FE, Paster BJ, Fox JG: Isolation and characterization of a Helicobacter sp. from the gastric mucosa of dolphins, Lagenorhynchus acutus and Delphinus delphis. Appl J Environ Microbiol, 66, 4751-4757, 2000.
11. Heilmann KF, Borchard F: Gastritis due to spiral shaped bacteria other than Helicobacter pylori: Clinical, histological and ultrastructural findings. Gut, 32, 137-140, 1991.
12. Andersen LP: New Helicobacter species in humans. Digest Dis J, 19, 112-115, 2001.
13. Happonen I, Saari S, Castren L, Tyni O, Hanninen, ML, 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.
14. Cattoli G, van Vugt RV, Zanoni RG, Sanguinetti V, Chiocchetti R, Gualtieri M, Vandenbroucke-Grauls CMJE, Gaastra W, Kusters JG: Occurrence and characterization of gastric Helicobacter spp. in naturally infected dogs. J Vet Microbiol, 70, 239-250, 1999.
15. Buczolits S, Hirt R, Rosengarten R, Busse HJ: PCR-based genetic evidence for occurrence of Helicobacter pylori and novel Helicobacter species in the canine gastric mucosa. J Vet Microbiol, 95, 259-270, 2003.
16. Ettinger SJ: Therapy of Arrhythmias. In, Ettinger SJ, Feldman EC (Eds): Text Book of Veterinary Internal Medicine. 7 th ed., 1321-1326, Saunders St Louis, 2010.
17. Shabestari Asl A, Jamshidi SH, Mohammadi M, Sasani F, Bahadori A, Oghalaie A: Assessment of chronic gastritis in pet dogs and its relation with Helicobacter-like organisms. Pakistan J Biol Sci, 11, 1443- 1448, 2008.
18. Shabestari Asl A, Jamshidi SH, Mohammadi M, Soroush MH, Bahadori A, Oghalaie A: Assessment of antimicrobial resistance of cultivable Helicobacter-like organisms in asymptomatic dogs. Iran J Vet Res, 10, 241-249, 2009.
19. Shabestari Asl, A, Jamshidi SH, Mohammadi M, Soroush MH, Bahadori A and Oghalaie A: Detection of atypical cultivable canine gastric Helicobacter strain and its biochemical and morphological characters in naturally ınfected dogs. Zoonoses Pub Health , 57, 244-248, 2010.
20. Van den Bulck KV, Decostere A, Baele M, Driessen A, Debongnie JC, Burette A, Stolte M, Ducatelle R, Haesebrouck F: Identification of non-Helicobacter pylori spiral organisms in gastric samples from humans, dogs and cats. J Clin Microb, 43, 2256-2260, 2005.
21. Stofel MH, Friess AE, Burnens A, Schmassmann A, Neiger R: Distinction of gastric Helicobacter spp. in human and domestic pets by scanning electron microscopy. Helicobacter, 5, 232-239, 2000.
22. Germani Y, Dauga C, Duval P, Huerre M, Levy M, Pialoux G, Sansonetti P, Grimont PAD: 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, 315326, 1997.
23. Neiger R, Dieterich C, Burnens A, Waldvogel A, Corthe´sy-Theulaz I, Halter F, Lauterburg B, Schmassmann A: Detection and prevalence of Helicobacter infection in pet cats. J Clin Microbiol, 36, 634637, 1998.
24. Labigne A, Cussac V, Courcoux P: Development of genetic and molecular approaches for the diagnosis and study of the pathogenicity of Helicobacter pylori, agent of gastric infammatory diseases. Bull Acad Natl Med, 175, 791-800, 1991.
25. Thompson MA, Strorey P, Greer R, Cleghorn J: Canine human transmission of Gastrospirillum hominis. Lancet, 343, 1605-1607, 1994.
26. Hanninen ML, Happonen I, Saari S, Jalava K: Culture and characteristics of Helicobacter bizzozeronii, a new canine gastric Helicobacter spp. Int J Syst Bacteriol, 46, 160-166, 1996.
27. Jalava K, On SLW, Vandamme PAR: Isolation and identification of Helicobacter spp. from canine and feline gastric mucosa. Appl J Environ Microbiol, 64, 3998-4006, 1998.
28. Van den Bulck K, Decostere A, Baele M, Vandamme P, Mast J, Ducatelle R, Haesebrouck F: Helicobacter cynogastricus sp. Nov., isolated from the canine gastric mucosa. Intl J Syst Evol Microbiol, 56, 1559-1564, 2006.