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Hayxoeuu eicnuK flHYBMET iMeni C.3. f^uцbкого
Tom 10 № 3(38) Hacmuna 1, 2008
Krzysztof Kubiak, Marcin Jankowski, Jolanta Spuzak, Jozef Nicpon, Grazyna Gosciniak*, Jacek Skala**, Urszula Paslawska, Kamila Glinska-Suchocka,
Vasyl Vlizlo*** ®
Department of Internal and Parasitic Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life
Sciences, Poland
*Department of Microbiology Medical University of Wroclaw, Poland **Institute of Genetics and Microbiology University of Wroclaw, Poland ***Institute of the Animal Biology of UAAS, Lviv, Ukraine
DIAGNOSIS OF HELICOBACTER SPP. INFECTION IN THE DOGS
STOMACH
Key words: Helicobacter spp., dog, stomach
Introduction
The infections with Helicobacter spp. in the stomach of dogs and cats are popular. Those microorganisms were isolated from the gastric mucosa of the healthy animals as well as from the mucosa of the affected stomach. The bacteria were found both in the cardiac part, the fundus, the corpus and the pyloric part of the stomach. Recently more and more authors claim that dogs and cats may be a reservoir of those microorganisms and constitute the source of infection for humans. So far the significance of the infections with Helicobacter spp. in the course of gastric diseases in dogs and cats has not been elucidated. The current knowledge of the subject is not complete and the problem calls for further research (1, 2, 3).
Task, the aim of the article
The aim of the study was to determine the frequency of occurrence of Helicobacter spp. microorganisms in the dogs stomach and the application and assessment of usefulness of invasive methods in the detection of stomach infections with Helicobacter spp.
Material and methods
The study entailed 157 dogs which were divided into two groups I and II.
The group I (control) included healthy animals in which no disorders in the functioning of the organism were found on the basis of anamnesis, clinical examination and the results of the hematological and biochemical blood tests. This group included also healthy animals in which the endoscopic examination (macroscopic assessment) did not show any lesions in gastric mucosa considered normotypical.
® Krzysztof Kubiak, Marcin Jankowski, Jolanta Spuzak, Jozef Nicpon, Grazyna Gosciniak, Jacek Skala, Urszula Paslawska, Kamila Glinska-Suchocka, Vasyl Vlizlo, 2008
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Науковий вгсник ЛНУВМБТ 1мен1 С.З. Гжицького Том 10 № 3(38) Частина 1, 2008
The group II (study) included dogs with dyspeptic signs indicating stomach diseases (vomiting of different character, decreased appetite, loss of appetite, changeable appetite, progressing body weight loss, fetor ex ore, painfulness in the stomach area). This group included also animals not manifesting the above clinical signs but in which the gastroscopy showed macroscopic lesions in gastric mucosa indicating gastritis. The group I included 20 dogs of different breed and sex, aged 6 months - 17 years; II - 137 dogs of different breed and sex, aged 2 months - 17 years. All the animals underwent gastroscopy during which mucosa bioptates were collected from the corpus and the pyloric part of the stomach for further examinations: microbiological ( rapid urease test, direct bacteriological preparation, culture examinations), PCR, histopathological ( hematoxyline and eosine staining and Giemza methods) and electron microscopy (scanning and transmission microscopy). The results were subjected to statistical analysis.
Results of research
The positive results of the rapid urease test were obtained in 18 (90%) dogs from the group I and 117 (85.4%) dogs from the group II.
The spiral, curvy, Gramm - negative microorganisms were found in the direct bacteriological preparation made from corpus mucosa samples in 16(80%) dogs from the group I and 107 (78.1%) dogs from the group II.
The spiral, curvy, Gramm - negative microorganisms were found in the direct bacteriological preparation made from pyloric mucosa samples in 18(90%) dogs from the group I and 94(68.6%) dogs from the group II.
On the basis of the bacteriological culture examinations Helicobacter spp. were found in 6(30%) dogs from the group I and in 21 (15.3%) dogs from the group II.
Basing on molecular PCR examinations the microorganisms of Helicobacter were found in gastric mucosa samples: in 20 (100%) dogs in the group I and in 133 (97.1%) dogs in the group II.
On the basis of the histopathological examinations of gastric bioptates from the corpus and the pylorus the chronic gastritis was observed in 3 (15%) dogs in the group I and in all the animals in the group II.
The histopathological examinations of gastric bioptates stained by Giemza method showed microorganisms of Helicobacter morphology in 5 (25%) dogs in the group I and in 36 (26.3%) dogs in the group II.
PCR examination of gastric bioptates in the examined population of animals showed Helicobacter heilmannii in 133 dogs, Helicobacter pylori - in 8, Helicobacter felis - in 81 and Helicobacter salomonis - in 6.
Conclusions
The conducted research showed that the infection of gastric mucosa with Helicobacter spp. is common in dogs and it concerns healthy and unhealthy animals. The most frequent species colonizing gastric mucosa in dogs is Helicobacter heilmannii. It was also proved that gastric mucosa in dogs may be colonized by
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Tom 10 № 3(38) Hacmuna 1, 2008
Helicobacter pylori. The examination of gastric mucosa bioptates by PCR method is the best way to detect the infection with Helicobacter spp. in dogs. The combined application of the classical PCR method, nested - PCR and seminested PCR with the use of appropriate primers allows the best species identification of those bacteria.
Reference
1. Neiger R., Tschudi M.E., Burnens A., Goke B., Schmassmann A.: 1999. Diagnostic and identification of gastric Helicobacter species by polymerase chain reaction in dogs. Microbiol. Ecol. Health. Dis. 11, 234-240.
2. Simpson K.W., Burrows C.F.: 1997. Zapalenie i choroba wrzodowa zol^dka, a zakazenie Helicobacter spp. u ludzi, psow i kotow. Waltham Focus. 7, 1-5.
3. Sapierzynski R., Malicka E.: 2004. Zakazenie Helicobacter spp. u kotow, psow i fretek. Mag. Wet. 13 (10), 14-16.
Summary
The aim of the study was to determine the frequency of occurrence of Helicobacter spp. microorganisms in the dogs stomach and the application and assessment of usefulness of invasive methods in the detection of stomach infections with Helicobacter spp. The conducted research showed that the infection of gastric mucosa with Helicobacter spp. is common in dogs and it concerns healthy and unhealthy animals. The most frequent species colonizing gastric mucosa in dogs is Helicobacter heilmannii. It was also proved that gastric mucosa in dogs may be colonized by Helicobacter pylori. The examination of gastric mucosa bioptates by PCR method is the best way to detect the infection with Helicobacter spp. in dogs. The combined application of the classical PCR method, nested - PCR and seminested PCR with the use of appropriate primers allows the best species identification of those bacteria.
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