Visualization of <i>Helicobacter</i> Species Within the Murine Cecal Mucosa Using Specific Fluorescence In Situ Hybridization

Chan, Vivian; Crocetti, Gregory; Grehan, Martin; Zhang, Li; Danon, Stephen J.; Lee, Adrian V.; Mitchell, Hazel M.

doi:10.1111/j.1523-5378.2005.00298.x

Cited by 34 publications

(34 citation statements)

References 32 publications

(40 reference statements)

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“…(Fuso0664),31 Enterobacteriaceae (Ebac1790),32 Helicobacter spp. (Hel717),33 and Faecalibacterium spp. (Faecali698),34 were labeled with Cy‐3 and were applied simultaneously with the universal bacterial probe Eub338‐FITC.…”

Section: Methodsmentioning

confidence: 99%

Relationship of the mucosal microbiota to gastrointestinal inflammation and small cell intestinal lymphoma in cats

Garraway

Johannes

Bryan

et al. 2018

Veterinary Internal Medicne

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BackgroundThe gastrointestinal (GI) microbiota in healthy cats is altered in IBD. Little research has been performed to identify whether specific bacterial groups are associated with small cell GI lymphoma (LSA).HypothesisMucosal bacteria, including Enterobacteriaceae and Fusobacterium spp., are abundant in intestinal biopsies of cats with small cell GI LSA compared to cats with IBD.AnimalsFourteen cats with IBD and 14 cats with small cell GI LSA.MethodsRetrospective case control study. A search of the medical records was performed to identify cats diagnosed with IBD and with GI LSA. Bacterial groups identified by FISH in GI biopsies were compared between cohorts and correlated to CD11b+ and NF‐κB expression.Results Fusobacterium spp. (median; IQR bacteria/region) were higher in cats with small cell GI LSA in ileal (527; 455.5 – 661.5; P = .046) and colonic (404.5; 328.8 – 455.5; P = .016) adherent mucus, and combined colonic compartments (free mucus, adherent mucus, attaching to epithelium) (8; 0 – 336; P = .017) compared to cats with IBD (ileum: 67; 31.5 – 259; colon: 142.5; 82.3 – 434.5; combined: 3; 0 – 34). Bacteroides spp. were higher in ileal adherent mucus (P = .036) and 3 combined ileal compartments (P = .034) of cats with small cell GI LSA. There were significant correlations between Fusobacterium spp. totals and CD11b+ cell (P = .009; rs .476) and NF‐κB expression (P = .004; rs .523).ConclusionsThe bacterial alterations appreciated might be influential in development of small cell GI LSA, and should drive further studies to elucidate the effects of microbial‐mediated inflammation on GI cancer progression.

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Section: Methodsmentioning

confidence: 99%

Relationship of the mucosal microbiota to gastrointestinal inflammation and small cell intestinal lymphoma in cats

Garraway

Johannes

Bryan

et al. 2018

Veterinary Internal Medicne

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“…although histological methods such as immunohistochemistry or fluorescence in situ hybridization (FISH) are options for detection of Helicobacter species to visualize morphologically intact microorganisms and their spatial distribution within the gastrointestinal tract [4,23], these methods are laborious and suffer from cross-reactivity with closely related bacteria such as Campylobacter species. there is however a positive correlation between the results of PCr detection of the 16s rrna gene and detection of organisms by immunohistochemistry or FISH [23].…”

Section: Detection Of Multiple Helicobacter Spp In An Mit 01-6451-inmentioning

confidence: 99%

“…to detect Helicobacter species infections, serologic tests, immunohistochemistry, and polymerase chain reaction (PCR) have been used [4,21,23,33]. Culture-based diagnostic tests are limited since, in general, Helicobacter species are slow-growing bacteria requiring more than 3 days of incubation for visible colony formation in a microaerobic atmosphere and culture conditions vary for different species [24,27].…”

Section: Introductionmentioning

confidence: 99%

Prevalence of an Unidentified <i>Helicobacter</i> Species in Laboratory Mice and its Distribution in the Hepatobiliary System and Gastrointestinal Tract

Yamanaka

Arita

et al. 2013

Exp. Anim.

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An unidentified Helicobacter species, strain MIT 01-6451, was frequently detected in mice obtained from domestic commercial and academic institutions in Japan. To partially characterize this strain, its distributions in the gastrointestinal tract and hepatobiliary system of mice were investigated. In gastrointestinal tissues, this strain was detected in all cecum, colon, and feces samples tested, whereas fewer mice were positive in the ileum, jejunum, and duodenum. Interestingly, strain MIT 01-6451 was also detected in most stomach samples and in 33% of gallbladder samples. One mouse was found to be infected with multiple Helicobacter species. Fourteen copies of 16S rRNA genes were cloned from the tissues of this mouse. One had the highest level of sequence homology with H. canadensis, while 13 had the highest level of homology with the H. ganmani type strain or strain MIT 01-6451. Twelve of these 13 16S rRNA genes were mosaic sequences, being partially derived from H. ganmani and strain MIT 01-6451. These results suggest that H. ganmani and Helicobacter sp. MIT 01-6451 are prevalent in specific-pathogen-free mouse colonies in Japan and that lateral gene transfer probably occurs among Helicobacter species during coinfection.

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“…genus were used to detect the novel Helicobacter sp. in the paraffin embedded intestinal tissues (26). Briefly, paraffin sections of ceca or colons from H.japonicum infected IL10 −/− mice were deparaffinized and rehydrated.…”

Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

“…Briefly, paraffin sections of ceca or colons from H.japonicum infected IL10 −/− mice were deparaffinized and rehydrated. A combination of two probes, HEL274 and HEL717, labeled with Cy3 were used to increase the intensity of the signals (Integrated DNA Technologies, Coralville, IA) (26). Hybridization buffer (0.9 M NaCl, 20 mM Tris-HCl, 0.01% SDS, 30% formamide) with 5 ng/ml of each probe was preheated for 10 min at 74.5 °C; 80 ul of this solution was added to each slide.…”

Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

Shen¹,

Feng²,

Muthupalani³

et al. 2016

CARCIN

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A novel Helicobacter species Helicobacter japonicum was isolated from the stomach and intestines of clinically normal mice received from three institutes from Japan. The novel Helicobacter sp. was microaerobic, grew at 37°C and 42°C, was catalase and oxidase positive, but urease negative. It is most closely related to the 16S rRNA gene of H.muridarum (98.6%); to the 23S rRNA gene of H.hepaticus (97.9%); to the hsp60 gene of H.typhlonius (87%). The novel Helicobacter sp. has in vitro cytolethal distending toxin (CDT) activity; its cdtB gene sequence has 83.8% identity with that of H.hepaticus. The whole genome sequence of H.japonicum MIT 01-6451 has a 2.06-Mb genome length with a 37.5% G + C content. When the organism was inoculated into C57BL/129 IL10 −/− mice, it was cultured from the stomach, colon and cecum of infected mice at 6 and 10 weeks post-infection. The cecum had the highest H.japonicum colonization levels by quantitative PCR. The histopathology of the lower bowel was characterized by moderate to severe inflammation, mild edema, epithelial defects, mild to severe hyperplasia, dysplasia and carcinoma. Inflammatory cytokines IFNγ, TNFα and IL17a, as well as iNOS were significantly upregulated in the cecal tissue of infected mice. These results demonstrate that the novel H.japonicum can induce inflammatory bowel disease and carcinoma in IL10 −/− mice and highlights the importance of identifying novel Helicobacter spp. especially when they are introduced from outside mouse colonies from different geographic locations.

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Visualization of Helicobacter Species Within the Murine Cecal Mucosa Using Specific Fluorescence In Situ Hybridization

Abstract: We have developed FISH probes specific for the family Helicobacteraceae as well as two individual Helicobacter species. This study will allow the future use of the FISH to better understand host-pathogen interactions in vitro.

Cited by 34 publications

References 32 publications

Relationship of the mucosal microbiota to gastrointestinal inflammation and small cell intestinal lymphoma in cats

Relationship of the mucosal microbiota to gastrointestinal inflammation and small cell intestinal lymphoma in cats

Prevalence of an Unidentified <i>Helicobacter</i> Species in Laboratory Mice and its Distribution in the Hepatobiliary System and Gastrointestinal Tract

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

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Visualization of Helicobacter Species Within the Murine Cecal Mucosa Using Specific Fluorescence In Situ Hybridization

Abstract: We have developed FISH probes specific for the family Helicobacteraceae as well as two individual Helicobacter species. This study will allow the future use of the FISH to better understand host-pathogen interactions in vitro.

Cited by 34 publications

References 32 publications

Relationship of the mucosal microbiota to gastrointestinal inflammation and small cell intestinal lymphoma in cats

Relationship of the mucosal microbiota to gastrointestinal inflammation and small cell intestinal lymphoma in cats

Prevalence of an Unidentified <i>Helicobacter</i> Species in Laboratory Mice and its Distribution in the Hepatobiliary System and Gastrointestinal Tract

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10−/−mice

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NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice