Detection of Helicobacter felis in a cat with gastric disease in laboratory animal facility / 한국실험동물학회지

A 3-month-old male cat in the animal facility was presented for investigation of anorexia and occasional vomiting. We collected the specimens from gastroscopic biopsy and stool collection. The gastroscopic biopsy specimens were tested using a rapid urease test, CLO Helicobacter-detection kits. Stool specimens were gathered and evaluated using the commercially available SD Bioline H. pylori Ag kit according to the manufacturer's instructions. Genomic DNAs from gastroscopic biopsy and stool specimens of the cat were extracted and submitted to the consensus PCR to amplify Helicobacter rpoB gene. Then the DNAs from gastroscopic biopsy and stool specimens were conducted a multiplex species-specific PCR to amplify urease B gene for H. heilmannii, H. pylori and H. felis. As the results, the rapid urease test with gastroscopic biopsy was revealed positive reaction. The result of H. pylori Stool Ag assay was one red line, negative for H. pylori. The gastroscopic biopsy and stool specimen were positive reactions by the consensus PCR reaction using the RNA polymerase beta-subunit-coding gene (rpoB) to detect Helicobacter species. By multiplex species-specific PCR with gastroscopic biopsy and stool specimens, no amplification products corresponding to either H. heilmannii or H. pylori were detected, but the specimens tested were positive for H. felis. This case was confirmed as gastroenteric disease induced by H. felis infection. On our knowledge, this is a very rare report about H. felis-induced gastroenteric disease in cat and may provide a valuable data on the study of feline Helicobacter infection.

Effect of N-Methyl-N-Nitrosourea on Helicobacter-induced Gastric Carcinogenesis in C57BL/6 Mice

BACKGROUND: The aim of this study was to investigate the effect of N-methyl-N-nitrosourea (MNU) treatment followed by chronic Helicobacter pylori SS1 and H. felis colonization on the stomachs of C57BL/6 mice. The role of MNU and Helicobacter species in gastric carcinogenesis was also elucidated. METHODS: A total of 69 C57BL/6 mice at 4 weeks of age were divided into 6 groups according to MNU treatment and H. pylori SS1 or H. felis infection. The mice were sacrificed at 21 and 50 weeks. The degree of inflammation was determined by histopathology. The levels of gastric mucosal myeloperoxidase, TNF-α, and interleukin-1β (IL-1β) were measured by ELISA. RESULTS: In the H. felis groups with or without MNU, the incidence of gastric tumors was 21.1% and 35.0% at 21 and 50 weeks, respectively. No gastric tumors were observed in all control mice. At 50 weeks, 37.5% of gastric adenoma cases were observed in the H. felis alone and MNU + H. felis groups. Furthermore, 12.5% of gastric adenocarcinoma cases were observed in the MNU alone and MNU + H. felis groups. The gastric mucosal IL-1β level was significantly higher in the MNU + H. felis group at 21 weeks and H. felis group at 50 weeks, respectively, than that for control mice (P < 0.05). However, the effect of MNU on H. pylori SS1-induced gastric carcinogenesis was low compared to that on H. felis. CONCLUSIONS: Administration of MNU before H. felis infection provokes severe inflammation through IL-1β, and eventually induces gastric cancer. However, the role of MNU in H. pylori SS1-induced gastric carcinogenesis model is minor.

Anti-inflammatory and Anti-tumorigenic Effects of Açai Berry in Helicobacter felis-infected mice

BACKGROUND: The aim of this study was to evaluate the anti-inflammatory and anti-tumorigenic effect of açai berry after chronic Helicobacter felis colonization in the stomachs of C57BL/6 mice. METHODS: A total of 57 four-week-old female C57BL/6 mice (18 control mice and 39 experimental mice) were used. The mice were administered orogastrically with vehicle only or vehicle containing H. felis, 5 times every other day. After inoculation of H. felis, mice were fed either a standard or an açai-containing diet and then sacrificed at 4, 24, and 52 weeks. The infection status and degree of inflammation were determined by culture and histopathology. The level of gastric mucosal myeloperoxidase (MPO), TNF-α, and interleukin-1β (IL-1β) were measured by ELISA. RESULTS: At 24 weeks after inoculation, mucosal atrophy and mucous metaplasia appeared in all infected mice. At 52 weeks after inoculation, dysplastic change was noted in 10%, 25%, and 50% of mice in the H. felis-control, H. felis-açai 5%, and H. felis-açai 10% groups, respectively. The neutrophil, monocyte, atrophy, and metaplasia grades of infected mice showed no significant difference among the H. felis-infected groups. H. felis-infected mice fed with açai berry showed no significant difference compared with H. felis-infected control mice in gastric mucosal MPO, TNF-α, and IL-1β levels. CONCLUSIONS: H. felis that colonized the stomachs of C57BL/6 mice provoked inflammation, and induced mucosal atrophy, metaplasia, and dysplasia. However, açai berry did not effectively prohibit the gastric carcinogenesis which was induced by chronic H. felis infection.

Comparison of three diagnostic assays for the identification of Helicobacter spp. in laboratory dogs / 한국실험동물학회지

A number of Helicobacter species may confound experimental data because of their association with disease progressing in various kinds of laboratory animals. Screening of Helicobacter species is particularly desirable, because they are prevalent in commercial and research animal facilities. The aim of the present study was to compare three diagnostic methods [e.g. Helicobacter stool antigen kit (HpSA), polymerase chain reaction (PCR) and rapid urease test (RUT)] for the identification of Helicobacter spp. in stools or gastric biopsy specimens collected from eight dogs suffering from gastritis. The gastroscopic biopsy specimens were tested using RUT and PCR, while stool specimens were evaluated using both HpSA and PCR. DNAs from the gastric biopsies and stool specimens were analyzed by both a consensus PCR that amplified the RNA polymerase beta-subunit-coding gene (rpoB) of Helicobacter spp. and a species-specific PCR to amplify the urease B gene of Helicobacter heilmannii, Helicobacter pylori, and Helicobacter felis. Helicobacter spp. were detected in 62.5% of the dogs, while H. heilmannii and H. felis were identified in 37.5 and 25% of the dogs, respectively. The HpSA did not efficiently detect Helicobacter spp. in the stool samples compared to the RUT and PCR assays, both of which successfully detected Helicobacter spp. in the two sample types. Finally, we recommend that consensus PCR with stool specimens could be used before the species-specific PCR for identifying Helicobacter species in laboratory dogs.

Evaluation of assays to detect Helicobacter felis infection in cats / 동물의과학연구지

Recently, several companies have released H. pylori stool antigen (HpSA) test kits. However, there is little information about the usefulness of HpSA testing for Helicobacter felis, which is the major Helicobacter species in cats. The aim of the present study was to compare diagnostic methods for diagnosis of H. felis with HpSA tests and PCR assay using cat stools or gastric mucosa. Male cats (n=6) were infected with H. felis ATCC 49179 (1.0 x 10(9) CFU /cat) by intragastric inoculation two times at 3-day intervals, and stool specimens of cats were collected 1, 3, 5, 7, 14, and 21 days after infection for HpSA testing and H. felis-specific PCR. For the results, sensitivities of the HpSA test and PCR analysis were 50.0% and 83.3% respectively. Cats were sacrificed 21 days after H. felis inoculation, and gastric tissues were homogenized. All gastric biopsy specimens were positive based on a rapid urease test (RUT) (6/6, 100%) and PCR (6/6, 100%). Based on these results, the HpSA kit is useful and effective for monitoring H. felis infection using stool specimens. If an HpSA test could be made with H. felis antibodies in the future, its sensitivity could be increased further. Further, PCR assay could be successfully used to detect H. felis in stools. Application of this HpSA kit and PCR assay can be utilized as a non-invasive strategy to identify H. felis in cats.

Gastric Helicobacter spp. infection in captive neotropical Brazilian feline

Ten captive neotropical Brazilian feline were submitted to gastroscopic examination and samples of gastric mucosa from fundus, corpus and pyloric antrum were evaluated for the presence of Helicobacter species. Warthin-Starry (WS) staining and PCR assay with species-specific primers and enzymatic cleavage were applied for bacterial detection and identification. Histological lesions were evaluated by haematoxylin and eosin staining. All animals showed normal gross aspect of gastric mucosa. Helicobacter heilmannii was confirmed in 100 percent of the samples by WS and PCR assay. Mild lymphocytic infiltrate in the lamina propria was observed in eight animals, mainly in the fundus region. Small lymphoid follicles were seen in three animals. No significant association between Helicobacter infection and histological findings was verified. These observations suggest that gastric Helicobacter spp. could be a commensal or a eventual pathogen to captive neotropical feline, and that procedures, way life, and stress level on the shelter apparently had no negative repercussion over the integrity of the stomach.

Postpartum Transmission as a Major Route of Mother-to-Child Helicobacter felis Infection / 한국실험동물학회지

In this study we investigated maternal Helicobacter felis (H. felis) infection status to determine the potential of maternal transmission. Pregnant Beagle dogs were infected experimentally with H. felis. Following the experimental design, the stools of the mother and litters were isolated and assessed for transmission of H. felis at parturition day, 1-week old age and 6-week old age respectively. Polymerase chain reaction (PCR) was used to examine the presence of transmitted H. felis. All litters showed no transmission of H. felis at parturition day. However, they revealed 14.3% and 100% at 1-week old age and 6-week old age respectively by PCR. These results suggested that vertical infection during prenatal period or delivery procedure is unlikely as a route of mother-to-child H. felis infection. It might be acquired H. felis through breast-feeding, contaminating saliva and fecal-oral during co-habitat.

Postpartum Transmission as a Major Route of Mother-to-Child Helicobacter felis Infection / 한국실험동물학회지

In this study we investigated maternal Helicobacter felis (H. felis) infection status to determine the potential of maternal transmission. Pregnant Beagle dogs were infected experimentally with H. felis. Following the experimental design, the stools of the mother and litters were isolated and assessed for transmission of H. felis at parturition day, 1-week old age and 6-week old age respectively. Polymerase chain reaction (PCR) was used to examine the presence of transmitted H. felis. All litters showed no transmission of H. felis at parturition day. However, they revealed 14.3% and 100% at 1-week old age and 6-week old age respectively by PCR. These results suggested that vertical infection during prenatal period or delivery procedure is unlikely as a route of mother-to-child H. felis infection. It might be acquired H. felis through breast-feeding, contaminating saliva and fecal-oral during co-habitat.

Survey of Helicobacter infection in domestic and feral cats in Korea

Discovery of Helicobacter (H.) pylori has led to a fundamental change in our understanding of gastric diseases in humans. Previous studies have found various Helicobacter spp. in dogs and cats, and pets have been questioned as a zoonotic carrier. The present study surveyed the Helicobacter infections and investigated the presence of H. felis and H. pylori infections in domestic and feral cats in Korea. Sixty-four domestic cats and 101 feral cats were selected from an animal shelter. Saliva and feces were evaluated by Helicobacter genus-specific polymerase chain reaction (PCR). Genus-specific PCR positive samples were further evaluated for H. felis and H. pylori using specific primer pairs. Thirty-six of 64 (56.3%) samples from domestic cats and 92 of 101 (91.1%) samples from feral cats were PCR positive; the positive rate of feces samples was higher than that of saliva samples in both groups. H. felis and H. pylori species-specific PCR was uniformly negative. The prevalence of Helicobacter spp. in feral cats was approximately two-fold higher than that of domestic cats. The fecal-oral route may be more a common transmission route not only between cats but also in humans.