Dogs are a natural reservoir for many of these species. Close and intense human contact with animals has been identified as a risk factor and therefore, an important zoonotic significance has been attributed to NHPH.
To determine the prevalence of Helicobacter species and the gastric histopathological changes associated, gastric mucosa samples of 69 dogs were evaluated. Only one dog presented a normal histopathological mucosa with absence of spiral-shaped organisms. A normal gastric mucosa and the presence of spiral-shaped bacteria was observed in two dogs. All remaining animals presented histopathological changes representative of gastritis.
Helicobacter species were detected in 60 dogs Histological, histochemical and immunohistochemical evaluations revealed that Helicobacter spp. Spiral-shaped bacteria were detected by qPCR analysis in 33 It was concluded that, despite the high incidence and worldwide distribution of gastric NHPH in dogs, the presence of specific Helicobacter species may vary between geographic regions.
NHPH infections were significantly accompanied by mild to moderate intraepithelial lymphocyte infiltration and mild to moderate gastric epithelial injury, but a clear relationship between gastritis and Helicobacter infection could not be established. The genus Helicobacter is composed of at least 40 species [ 1 ].
Among these, H. A large number of non- Helicobacter pylori Helicobacter species NHPH have also been recognized in humans and in several animals. Most of these gastric NHPH are also able to cause disease in humans [ 6 , 7 ]. Close and intense human contact with animals has been identified as a risk factor and therefore, an important zoonotic significance has been attributed to NHPH [ 6 , 8 , 9 ].
In pet animals, gastric Helicobacter spp. The predominant gastric Helicobacter spp. Mixed infections with different species can also occur [ 3 , 19 ]. While many studies have reported that the fundus and body have higher bacterial density and a higher probability of finding Helicobacter spp. The discrepancies in these results can be attributed to the different laboratory diagnostic methodologies used by the various research groups. The diagnostic methods used for Helicobacter spp.
Non-invasive methods like serology or detection of bacterial DNA and antigens in stools do not require a gastric biopsy or anaesthesia. The invasive methods, like bacterial cultures, histopathology, smears, electron microscopy or polymerase chain reaction PCR may require a gastric biopsy, which is frequently obtained through endoscopy under anaesthesia or by necropsy. Typically Helicobacter organisms are not easily visualized with the haematoxylin and eosin HE stain and so, their direct observation in biopsied specimens is highlighted by the use of special stains, such as the modified Giemsa MG stain.
More elaborate and sensitive Helicobacter -detection methods such as immunohistochemistry IHC or polymerase chain reaction PCR are research tools rarely used in a diagnostic setting. Several investigations have discussed the prevalence of Helicobacter spp. The accurate identification of the gastric helicobacters to the species level is essential in order to determine the prevalence and clinical significance of all taxa.
The aim of this study was to determine the prevalence of different gastric Helicobacter species present in distinct stomach regions of the canine stomach body and antrum using histological, histochemical, immunohistochemical and molecular diagnostic techniques.
The degree of colonization was characterized and correlated with the respective histopathological changes in the canine gastric mucosa. Gastric tissues were obtained from 69 dogs 45 male and 24 female, ranging in age from 3 months to 15 years. Samples were obtained from 20 dogs during endoscopic procedures, from five during surgery and from 44 dogs during necropsy examinations.
All the procedures surgical excision and necropsy examination were performed in a clinical context attempting to treat the animals based on the best clinical judgment of their attending practitioners. The use of the excised tissues for research was explained to the owners and an informed consent was obtained for each case. None of the actions were taken solely for research purposes and the investigators had no influence on the selection and execution of such procedures.
Only gastric samples in good condition of preservation were included in this study. The gastric samples were evaluated independently by two observers IA and FG. Histopathological parameters such as alterations in cellularity, fibrosis of the lamina propria and gland atrophy were analysed according to the World Small Animal Veterinary Association WSAVA guidelines [ 29 ]. The degree of morphological features and inflammatory changes was graded as normal, mild, moderate or marked by using the available WSAVA gastrointestinal standardization visual analogue [ 29 ].
The microscopic evaluation was performed by analysing the entire section of the gastric tissue. The presence of Helicobacter spp. A dog was classified as Helicobacter positive when one of these methods gave a positive result. Slides were cooled for 10 min at room temperature and rinsed twice in triphosphate buffered saline TBS for 5 min.
Sections were rinsed with TBS between each step of the procedure. Louis, MO and sections were then lightly counterstained with haematoxylin, dehydrated and mounted. Positive immunoreactivity was recorded as a distinct golden-brown labelling of the bacteria located on mucosal surface, in gastric pits or glands and in parietal cells. Helicobacter species-specific qPCRs based on a short fragment of the urease A and B genes were developed for the identification of H.
The average values of the duplicates were used for quantification of Helicobacter DNA in the tissue samples. To exclude false positive samples, the amplicons from each positive sample were sequenced.
Statistical analysis was performed using the statistical package SPSS A total of gastric samples 66 from the body region and 51 from the antrum region were analysed. Among the 69 animals, both gastric regions were available for evaluation in 48 dogs whereas only body or antrum regions were available in 18 and three dogs, respectively. Of the 69 dogs, only one presented a normal histopathological mucosa with absence of spiral-shaped organisms. A normal gastric mucosa and the presence of spiral-shaped bacteria was observed in two dogs 2.
On the basis of histopathological changes in the gastric mucosa, we diagnosed a mild to moderate chronic gastritis in Both mild to moderate epithelial injuries and mild to moderate intraepithelial lymphocyte infiltration were found in Gastric mucosal atrophy, glandular nesting or fibrosis was present in In Abnormal neutrophilic infiltration was only detected in the stomach antrum of two animals: in one, this alteration was mild and in the other, it was marked and associated with gastric ulceration.
Other inflammatory cells, consisting in mild infiltration of mast cells, were observed in the body region of four animals and in the antrum of two animals. Among all animals, Regardless of the stomach location, Helicobacter spp. With HE staining, spiral-shaped bacteria were detected in Utilizing the MG stain, this was Helicobacter antigen was detected by immunochemistry in Helicobacter spp.
A Numerous spiral-shaped bacteria colonizing the surface epithelium of the gastric pit. Inset shows Helicobacter antigen within parietal cells, sometimes detectable as well preserved spiral-shaped organisms or brown round dots. Further identification at species level was performed using Helicobacter species-specific qPCRs. The majority of the samples were positive in the H. Therefore, these cases were reclassified as H. Equal proportions of H.
Mixed infections with H. In the body area, the most frequently identified species was H. There was a significant correlation between the presence of Helicobacter spp. No statistically significantly correlations were found between Helicobacter infection and gastric mucosal atrophy or fibrosis, lamina propria lymphoplasmacytic infiltration or lymphofollicular hyperplasia.
In this study, a high prevalence of gastritis was observed As in human medicine, the mode of transmission is unclear. It is very possibly spread by oral ingestion of contaminated food and water, but oral transmission by ingestion of vomitus is more likely, especially in puppies.
When is Helicobacter spread? Most infections are acquired during childhood and a person can then carry the infection for life and never develop symptoms. What are the symptoms of Helicobacter? The prevalence of gastric Helicobacter spp. In dogs, Helicobacter has been tied to chronic inflammation of the lining of the stomach. However, studies have shown that untreated infections can result in significant inflammation.
The most common of these are discussed below. Gastrointestinal campylobacteriosis is a bacterial disease. It is caused by related It is caused by bacteria From developing new therapies that treat and prevent disease to helping people in need, we are committed to improving health and well-being around the world.
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