J Vet Sci.  2007 Jun;8(2):139-145. 10.4142/jvs.2007.8.2.139.

Diagnosis of Helicobacter spp. infection in canine stomach

• Prachasilpchai W
• Nuanualsuwan S
• Chatsuwan
• Techangamsuwan S
• Wangnaitham S
• Sailasuta

• KMID: 1089664
• DOI: http://doi.org/10.4142/jvs.2007.8.2.139

Abstract

A total of 75 biopsied samples of cardia, fundus, body, and pyloric antrum from necropsied dogs that were submitted to the Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University from April 2003 to June 2004 were investigated. The objectives of this study were to determine the prevalence of Helicobacter spp. in canine stomach by polymerase chain reaction (PCR) in comparison to histochemistry versus immunohistochemistry (IHC), and to correlate these diagnostic methods with the clinical significance in infected dogs. Histopathological results revealed 60.0% (45/75) of samples to be positive, and consisted of mild gastritis in 64.44% (29/45), moderate gastritis in 11.11% (5/45), and severe gastritis in 24.44% (11/45). The proportion showing no histopathological lesions was 40.0% (30/75). Helicobacter spp. were localized to the luminal crypt in 18.67% (14/75), gastric pit in 22.67% (17/75), gastric gland in 21.33% (16/75), and gastric epithelium in 8% (6/75). The percentages of positive samples of Helicobacter spp. diagnosed by hematoxylin and eosin stain (H&E), Warthin Starry stain (WSS), IHC with rabbit polyclonal anti-H. pylori antibody, and PCR were 17.3% (13/75), 46.7% (35/75), 30.7% (23/75), and 10.7% (8/75), respectively. No significant differences weree observed in histopathological changes in portions of the stomach (p > 0.05). The diagnosis of Helicobacter spp. by PCR in comparison to that by WSS and IHC was not significantly different (p > 0.05). There were no relationships between pathological studies using H&E, WSS, and IHC, and especially between PCR and clinical signs of Helicobacter spp. infections in canine stomachs (p > 0.05). The present study revealed significantly different levels of correlation for Helicobacter spp. detection between H&E and WSS (p < 0.001). Results indicate that the method of choice for diagnosis of Helicobacter spp. infection in canine stomach is dependent on the purpose of study and appropriate specimen collection.

Keyword

dog; Helicobacter spp.; stomach

MeSH Terms

Animals
DNA, Bacterial/chemistry/genetics
Dog Diseases/*diagnosis/epidemiology/*microbiology
Dogs
Female
Gastric Mucosa/microbiology
Gastritis/diagnosis/epidemiology/microbiology/*veterinary
Helicobacter/genetics/*isolation & purification
Helicobacter Infections/diagnosis/epidemiology/microbiology/*veterinary
Immunohistochemistry/veterinary
Male
Polymerase Chain Reaction/veterinary

Figure

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  Fig. 1 Macroscopic lesions were shown as chronic mutifocal ulcerative gastritis; a 0.5 × 1.5 cm ulcer was shown on the gastric mucosa in the body part of the stomach (green arrowhead).

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  Fig. 2 The histopathological results revealed the formation of multiple lymphoid follicles, which were numerous in gastric mucosa. H&E stain.

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  Fig. 3 The numerous spiral-shaped organisms: Helicobacter spp. is shown in the surface of gastric mucosa in the fundus of the stomach. H&E stain.

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  Fig. 4 Spiral-shaped Helicobacter spp. was positive, as indicated by a dark brown color, and was 3.0-5.0 µm in length and 0.5-1.0 µm in width in the gastric pit of the fundus. At a higher magnification, Helicobacter spp. with a spiral shape and dark brown color are shown. Warthin Starry stain.

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  Fig. 5 Helicobacter spp. showed a brown color in positive sites by immunohistochemical staining with rabbit polyclonal anti-H. pylori antibody in the lumen of the gastric gland in the fundus portion of the stomach. Counterstaining with Meyer's hematoxylin.

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  Fig. 6 The 400 bp bands were detected by 2% agarose gel electrophoresis following PCR amplification of the Helicobacter spp.-specific sequences. Lane 1-8: positive results from canine gastric tissues; Lane 9: positive control H. Pylori from human; Lane 10: negative control nuclease-free water; Lane 11: 100 bp DNA marker ladder.

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