A group-specific PCR assay for the detection of Helicobacteraceae in human gut

Background. Enterohepatic Helicobacter species are emerging pathogens, which are increasingly isolated from humans with enteric diseases. Nevertheless, current methods to detect Helicobacteraceae in the human gut have significant limitations. Methods. Based on 16S‐rRNA gene alignments and computer aided primer analysis a set of group‐specific PCR primers was designed. The evaluation of the PCR assay was performed using 36 ATCC reference strains and intestinal biopsies from 10 patients with defined gastric Helicobacter pylori status. The amplification products derived from clinical samples were cloned and subsequently analyzed by DNA sequencing. Sensitivity of the PCR‐assay was determined by spiking previously negative tested samples with decreasing amounts of Helicobacter DNA. Results. The analysis of the ATCC reference strains revealed amplification products in all 14 Helicobacter strains and Wolinella succinogenes, 21 other microorganisms representing negative controls did not produce PCR fragments. Four out of the 10 patient‐derived samples were positive. Three of them represented H. pylori‐derived DNA confirming the gastric H. pylori infection in these patients. In the fourth patient, who was suffering from Crohn's disease, H. pullorum was identified. The sensitivity of the PCR assay was 0.1 pg of Helicobacter‐derived DNA representing about 40 bacteria. Conclusion. The novel PCR assay described here is an important new tool in rapid and sensitive assessment for the presence of Helicobacteraceae in human gut.     

Detection of Helicobacteraceae in intestinal biopsies of children with Crohn's disease

Background: Given that members of Helicobacteraceae family colonize the intestinal mucus layer, it has been hypothesized that they may play a role in Crohn’s disease. This study investigated the presence of Helicobacteraceae DNA in biopsies collected from children with Crohn’s disease and controls. Materials and Methods: The presence of Helicobacteraceae DNA was investigated in intestinal biopsies collected from 179 children undergoing colonoscopy (Crohn’s disease n = 77, controls n = 102) using a Helicobacteraceae‐specific PCR. Results: Members of the Helicobacteraceae were detected in 32/77 children with Crohn’s disease (41.5%) and 23/102 controls (22.5%). Statistical analysis showed the prevalence of Helicobacteraceae detected in patients to be significantly higher than that in controls (p = .0062). Analysis of non‐pylori Helicobacteraceae showed that their prevalence was also significantly higher in patients than in controls (p = .04). Helicobacter pylori was detected in 14.0% of the biopsies across all groups. Given that all children tested were negative for gastric H. pylori, this was a surprising finding. Phylogenetic analysis of H. pylori sequences detected in the biopsies showed that the H. pylori strains identified in the patients did not group with gastric H. pylori included in the analysis, but rather with other H. pylori strains detected in the intestine, gall bladder, and liver. Conclusions:  The higher prevalence of Helicobacteraceae DNA in Crohn’s disease patients would suggest that members of this family may be involved in this disease. In addition, phylogenetic analysis of H. pylori strains showed that extragastric sequences clustered together, indicating that different H. pylori strains may adapt to colonize extragastric niches.     

Detection of Helicobacter Species in the Gastrointestinal Tract of Wild Rodents From Brazil

Since we have limited knowledge about the occurrence of Helicobacter in wild animals, we searched for Helicobacter species in the gastrointestinal tract of 75 rodents captured in forest remnants of Minas Gerais, Brazil. Fragments from the antrum and corpus of the stomach and from the colon were taken for PCR assays for Helicobacter detection. Although gastric mucosa was Helicobacter-positive in only one animal, the bacterium was detected in the colonic mucosa of 23 rodents (30.7%). Helicobacter detection was more frequent in the colon of terraced rice rat (56%) and house rat (30%) in contrast to punare and Spix’s yellow-toothed cavy, in which the presence of the bacterium was not detected. Helicobacter rodentium, H. marmotae, H. cinaedi, and other species closely related to the murine helicobacters were presumptively identified by DNA sequencing. Wild rodents may serve as a reservoir of these Helicobacter species in nature.     

Effects of myeloid differentiation primary response gene 88 (MyD88) activation on Helicobacter infection in vivo and induction of a Th17 response

Background:  Helicobacter pylori is a spiral‐shaped Gram‐negative microaerophilic bacterium associated with a number of gastrointestinal disorders, including gastritis, peptic ulcers, and gastric cancer. Several studies have implicated a Th17 response as a key to protective immunity against Helicobacter. Materials and Methods:  Wild type (WT) and MyD88‐deficient (MyD88^?/?) mice in the C57BL/6 background were infected with H. felis for 6 and 25 weeks and colonization density and host response evaluated. Real‐time PCR was used to determine the expression of cytokines and antimicrobial peptides in the gastric tissue of mice. Results:  mRNA expression levels of the Th17 cytokines interleukin‐17A (IL‐17A) and IL‐22 were markedly up‐regulated in WT compared with MyD88^?/? mice both at 6 and at 25 weeks in response to infection with H. felis, indicating that induction of Th17 responses depends on MyD88 signaling. Furthermore, reduction in the expression of Th17‐dependent intestinal antimicrobial peptide lipocalin‐2 was linked with increased bacterial burden in the absence of MyD88 signaling. Conclusion:  We provide evidence showing that MyD88‐dependent signaling is required for the host to induce a Th17 response for the control of Helicobacter infection.     

Morphologic, genetic, and biochemical characterization of Helicobacter magdeburgensis, a novel species isolated from the intestine of laboratory mice

Background: The presence of enterohepatic Helicobacter species (EHS) is commonly noted in mouse colonies. These infections often remain unrecognized but can cause severe health complications or more subtle host immune perturbations and therefore can confound the results of animal experiments. The aim of this study was to isolate and characterize a putative novel EHS that has previously been detected by PCR screening of specific‐pathogen‐free mice. Materials and Methods: Biochemical analysis of enzyme activities (API campy), morphologic investigation (Gram‐staining and electron microscopy) and genetic analyses (16SrRNA and 23SrRNA analyses, DNA fingerprinting, restriction fragment polymorphisms, and pulsed‐field gel electrophoresis) were used to characterize isolated EHS. Genomic DNA fragments were sequenced to develop a species‐specific PCR detection assay. Results: Scanning electron microscopy revealed the presence of spiral‐shaped EHS, which varied in length (2.5–6 μm) and contained single monopolar or single bipolar sheathed flagella. The bacteria were grown under anaerobic conditions, preferably on agar plates containing serum or blood. The 16SrRNA, genetic, and biochemical analyses indicated the identification of a novel EHS species, named Helicobacter magdeburgensis. We also examined the genome content using pulsed‐field gel electrophoresis. Based on the pattern produced by two restriction enzymes, BamIII and KspI, the genome size was determined to be about 1.7–1.8 Mbp. Conclusion: We isolated and characterized a novel EHS species, H. magdeburgensis, morphologically, biochemically, and genetically. These results are important for future studies on the prevalence and pathophysiologic relevance of such infections. Our PCR assay can be used to detect and discriminate H. magdeburgensis from other Helicobacter species.     

Recent advances in the understanding of Xanthomonas citri ssp. citri pathogenesis and citrus canker disease management

Taxonomic status : Bacteria; Phylum Proteobacteria; Class Gammaproteobacteria; Order Xanthomonadales; Family Xanthomonadaceae; Genus Xanthomonas; Species Xanthomonas citri ssp. citri (Xcc). Host range : Compatible hosts vary in their susceptibility to citrus canker (CC), with grapefruit, lime and lemon being the most susceptible, sweet orange being moderately susceptible, and kumquat and calamondin being amongst the least susceptible. Microbiological properties : Xcc is a rod‐shaped (1.5–2.0 × 0.5–0.75 µm), Gram‐negative, aerobic bacterium with a single polar flagellum. The bacterium forms yellow colonies on culture media as a result of the production of xanthomonadin. Distribution : Present in South America, the British Virgin Islands, Africa, the Middle East, India, Asia and the South Pacific islands. Localized incidence in the USA, Argentina, Brazil, Bolivia, Uruguay, Senegal, Mali, Burkina Faso, Tanzania, Iran, Saudi Arabia, Yemen and Bangladesh. Widespread throughout Paraguay, Comoros, China, Japan, Malaysia and Vietnam. Eradicated from South Africa, Australia and New Zealand. Absent from Europe.     

Reduction of Helicobacter Infection in IL-10−/– Mice is Dependent On CD4+ T Cells but not on Mast Cells

Background: In contrast to wild type, interleukin‐10‐deficient (IL‐10^?/–) mice are able to clear Helicobacter infection. In this study, we investigated the immune response of IL‐10^?/– mice leading to the reduction of Helicobacter infection. Materials and Methods: We characterized the immune responses of Helicobacter felis‐infected IL‐10^?/– mice by studying the systemic antibody and cellular responses toward Helicobacter. We investigated the role of CD4⁺ T cells in the Helicobacter clearance by injecting H. felis‐infected IL‐10^?/– mice with anti‐CD4 depleting antibodies. To examine the role of mast cells in Helicobacter clearance, we constructed and infected mast cells and IL‐10 double‐deficient mice. Results: Reduction of Helicobacter infection in IL‐10^?/– mice is associated with strong humoral (fivefold higher serum antiurease antibody titers were measured in IL‐10^?/– in comparison to wild‐type mice, p < .008) and cellular (urease‐stimulated splenic CD4⁺ T cells isolated from infected IL‐10^?/– mice produce 150‐fold more interferon‐γ in comparison to wild‐type counterparts, p < .008) immune responses directed toward Helicobacter. Depletion of CD4⁺ cells from Helicobacter‐infected IL‐10^?/– mice lead to the loss of bacterial clearance (rapid urease tests are threefold higher in CD4⁺ depleted IL‐10^?/– in comparison to nondepleted IL‐10^?/– mice, p < .02). Mast cell IL‐10^?/– double‐deficient mice clear H. felis infection, indicating that mast cells are unnecessary for the bacterial eradication in IL‐10^?/– mice. Conclusion: Taken together, these results suggest that CD4⁺ cells are required for Helicobacter clearance in IL‐10^?/– mice. This reduction of Helicobacter infection is, however, not dependent on the mast cell population.     

Case Report: Helicobacter suis Infection in a Pig Veterinarian

This study describes a non‐Helicobacter (H.) pylori Helicobacter (NHPH) infection in a pig veterinarian. The patient suffered from reflux esophagitis and general dyspeptic symptoms and was referred to the hospital for upper gastrointestinal endoscopy. Histologic examination of corpus and antrum biopsies revealed a chronic gastritis. Large spiral‐shaped non‐H. pylori helicobacters could be visualized and were identified as H. suis by PCR. The patient was treated with a triple therapy, consisting of amoxicillin, clarithromycin, and pantoprazole for 10 days. Successful eradication was confirmed after a follow‐up gastrointestinal endoscopy and PCR 10 weeks after treatment. A mild chronic gastritis was, however, still observed at this point in time. This case report associates porcine H. suis strains with gastric disease in humans, thus emphasizing the zoonotic importance of H. suis bacteria from pigs.     

Characterization and evolution of two bacteriome‐inhabiting symbionts in cixiid planthoppers (Hemiptera: Fulgoromorpha: Pentastirini)

Like other plant sap‐sucking insects, planthoppers within the family Cixiidae (Insecta: Hemiptera: Fulgoromorpha) host a diversified microbiota. We report the identification and first molecular characterization of symbiotic bacteria in cixiid planthoppers (tribe: Pentastirini). Using universal eubacterial primers we first screened the eubacterial 16S rRNA sequences in Pentastiridius leporinus (Linnaeus) with PCR amplification, cloning, and restriction fragment analysis. We identified three main 16S rRNA sequences that corresponded to a Wolbachia bacterium, a plant pathogenic bacterium, and a novel gammaproteobacterial symbiont. A fourth bacterial species affiliated with ‘Candidatus Sulcia muelleri’ was detected in PCR assays using primers specific for the Bacteroidetes. Within females of two selected cixiid planthoppers, P. leporinus and Oliarus filicicola, fluorescence In situ hybridization analysis and transmission electron microscopy observations showed that ‘Ca. Sulcia muelleri’ and the novel gammaproteobacterial symbiont were housed in separate bacteriomes. Phylogenetic analysis revealed that both of these symbionts occurred in at least four insect genera within the tribe Pentastirini. ‘Candidatus Purcelliella pentastirinorum’ was proposed as the novel gammaproteobacterial symbiont.     

Presence of gastric Helicobacter species in children suffering from gastric disorders in Southern Turkey

Background

Infections with gastric Helicobacter spp. are associated with gastritis, peptic ulceration, and malignancies. Helicobacter pylori is the most prevalent Helicobacter species colonizing the human stomach. Other gastric non‐H. pylori helicobacters (NHPHs) have been described in 0.2%‐6% of human patients with gastric disorders. Nevertheless, due to difficulties in the diagnosis of NHPH infections and lack of routine screening, this is most likely an underestimation of their true prevalence. To the best of our knowledge, no studies have been performed in the presence of Helicobacter spp. in children suffering from gastric disorders in Southern Turkey.

Materials and methods

In total, 110 children with gastric complaints were examined at the Cukurova University Balcali hospital, Turkey. Gastroscopy was performed to evaluate the presence of gastric mucosal lesions. Biopsies of the pyloric gland zone were taken for histopathological analysis, rapid urease testing, and presence of Helicobacter spp. DNA by PCR.

Results

Based on the PCR results, the prevalence of Helicobacter spp. was 32.7% (36/110). H. pylori was found in 30.9% (34/110), H. suis in 1.8% (2/110), and H. heilmannii/H. ailurogastricus in 0.9% (1/110) of the human patients. A mixed infection with H. pylori and H. suis was present in one patient. The presence of mucosal abnormalities, such as nodular inflammation, ulceration, and hyperemia, as well as gastritis, was significantly higher in Helicobacter spp. positive patients.

Conclusion

Helicobacter pylori, H. suis, and H. heilmannii/H. ailurogastricus were present in children with gastric complaints. Infection with these pathogens may be involved in the development of gastritis and ulceration.     

Succinate:quinone oxidoreductases from ϵ-proteobacteria

The ϵ-proteobacteria form a subdivision of the Proteobacteria including the genera Wolinella, Campylobacter, Helicobacter, Sulfurospirillum, Arcobacter and Dehalospirillum. The majority of these bacteria are oxidase-positive microaerophiles indicating an electron transport chain with molecular oxygen as terminal electron acceptor. However, numerous members of the ϵ-proteobacteria also grow in the absence of oxygen. The common presence of menaquinone and fumarate reduction activity suggests anaerobic fumarate respiration which was demonstrated for the rumen bacterium Wolinella succinogenes as well as for Sulfurospirillum deleyianum, Campylobacter fetus, Campylobacter rectus and Dehalospirillum multivorans. To date, complete genome sequences of Helicobacter pylori and Campylobacter jejuni are available. These bacteria and W. succinogenes contain the genes frdC, A and B encoding highly similar heterotrimeric enzyme complexes belonging to the family of succinate:quinone oxidoreductases. The crystal structure of the W. succinogenes quinol:fumarate reductase complex (FrdCAB) was solved recently, thus providing a model of succinate:quinone oxidoreductases from ϵ-proteobacteria. Succinate:quinone oxidoreductases are being discussed as possible therapeutic targets in the treatment of several pathogenic ϵ-proteobacteria.     

Rickettsia africae infection rates and transovarial transmission in Amblyomma hebraeum ticks in Mnisi,Bushbuckridge, South Africa

Rickettsia africae is a gram-negative bacterium, which causes African tick bite fever (ATBF) in humans. ATBF is a febrile disease mainly affecting travellers to southern Africa. This bacterium is known to be transmitted by Amblyomma hebraeum and Amblyomma variegatum ticks. In southern Africa, the principal vector is A. hebraeum. Febrile disease is a serious issue in the study area. There is a high prevalence of non-malaria illness caused by Rickettsia, so there is a need to have more knowledge on these species. Infection rates and transovarial transmission efficiency of R. africae in A. hebraeum ticks were investigated in a rural area of Mpumalanga province, South Africa. Adult and engorged A. hebraeum female ticks were collected from cattle. Larvae were collected by dragging a cloth at ground level using 100 steps, equivalent to an area of 100 m². Tick identification was performed according to standard taxonomic keys using a microscope. Engorged ticks were incubated to oviposit and egg masses were collected. DNA was extracted from the ticks, larvae and egg masses, and screened for gltA and ompA genes, using quantitative real-time PCR and conventional PCR, respectively. Positive ompA amplicons were sequenced and phylogenetic analysis showed 99.8-100% identity with R. africae. Infection rates were 13.7 and 12.7% for adults and larvae, respectively. Transovarial transmission of R. africae in A. hebraeum from this study was 85.7%. The results provide a clear indication that people living in the study area and travellers that visit the area are at risk of contracting ATBF.

     

Isolation and Characterization of Helicobacter Species from the Stomach of the House Musk Shrew (Suncus murinus) with Chronic Gastritis

A Gram-negative, motile bacterium with bipolar sheathed flagella (one at each end) was isolated from the stomach of house musk shrews (Suncus murinus) with chronic gastritis. The isolates grew at 37°C under microaerophilic conditions, but not under aerobic conditions; rapidly hydrolyzed urea; were catalase, oxidase, alkaline phosphatase, and arginine aminopeptidase positive; reduced nitrate to nitrite; and were resistant to cephalothin and nalidixic acid, but sensitive to tetracycline, erythromycin, and chloramphenicol. This bacterium was found on gastric epithelial cells by electron microscopy. In addition, a coccoid form of the bacteria was found in vacuoles formed in the epithelial cells of some of the house musk shrews tested. These results, including 16S rRNA gene sequence analysis, strongly suggested that this bacterium should be classified as a novel Helicobacter species. It is proposed that this bacterium should be called “Helicobacter suncus.” Received: 22 December 1997 / Accepted: 26 January 1998     

Male‐killer dynamics in Danaus chrysippus (L.) (Lepidoptera: Nymphalidae) in East Africa

Within certain regions in East Africa, the butterfly Danaus chrysippus (L.) shows female‐biased population sex ratio, because of the production by some females of all‐female broods, as a result of infection by maternally inherited, male‐killing bacterium of the genus Spiroplasma. In this study, we describe a 3‐year field survey for the population dynamics of the male‐killing Spiroplasma in D. chrysippus in four independent localities, namely Uganda, Ghana, Sudan and Madagascar. The prevalence of the bacterium was found to show extensive variations at multiple scales among different sites, in various countries, seasons and years. A novel, selection‐based hypothesis was suggested to explain the high variability of male‐killer prevalence over space and time, based on the existence of an adaptive link between larval food‐plant density and the magnitude of resource reallocation fitness advantage for the male‐killer.     

Widespread historical presence of Batrachochytrium dendrobatidis in African pipid frogs

Aim Amphibian chytridiomycosis, an emerging infectious disease caused by the chytrid fungus Batrachochytrium dendrobatidis (Bd), is associated with global amphibian population declines and species extinctions. Current evidence indicates that the pathogen has recently spread globally from an enzootic focus, with Xenopus spp. (family Pipidae) in South Africa having been identified as a likely source. The aim of this study was to investigate further the likelihood of African Xenopus spp. as the original source of Bd. Location We examined 665 museum specimens of 20 species of African and South American pipid frogs collected between 1844 and 1994 and held in the collection of the Natural History Museum, London. Methods Skin brushings taken from adult amphibians and brushings from the mouthparts, lips and developing hind limbs of larval pipid frogs were examined for the presence of Bd using real‐time PCR. Results We found six cases of Bd infection in three Xenopus spp. (from Africa), but none of the South American pipids was positive, although only 45 South American frogs were available for examination. The earliest case of Bd infection was in a specimen of Xenopus fraseri collected from Cameroon in 1933. A consistently low prevalence of infection over time indicates that a historical equilibrium existed between Xenopus spp. and Bd infection in Africa. Main conclusions Our results suggest that Bd infection was present in Xenopus spp. across sub‐Saharan Africa by the 1930s, providing additional support for the ‘out of Africa’ hypothesis. If this hypothesis is correct, it strengthens the argument for stringent control of human‐assisted movements of amphibians and other wildlife world‐wide to minimize the likelihood of pathogen introduction and disease emergence that can threaten species globally. Our findings help inform species selection for conservation in the face of the current Bd pandemic and also guide future research directions for selecting Bd isolates for sequencing and virulence testing.     

Patterns of selection on Plasmodium falciparum erythrocyte‐binding antigens after the colonization of the New World

Pathogens, which have recently colonized a new host species or new populations of the same host, are interesting models for understanding how populations may evolve in response to novel environments. During its colonization of South America from Africa, Plasmodium falciparum, the main agent of malaria, has been exposed to new conditions in distinctive new human populations (Amerindian and populations of mixed origins) that likely exerted new selective pressures on the parasite's genome. Among the genes that might have experienced strong selective pressures in response to these environmental changes, the eba genes (erythrocyte‐binding antigens genes), which are involved in the invasion of the human red blood cells, constitute good candidates. In this study, we analysed, in South America, the polymorphism of three eba genes (eba‐140, eba‐175, eba‐181) and compared it to the polymorphism observed in African populations. The aim was to determine whether these genes faced selective pressures in South America distinct from what they experienced in Africa. Patterns of genetic variability of these genes were compared to the patterns observed at two housekeeping genes (adsl and serca) and 272 SNPs to separate adaptive effects from demographic effects. We show that, conversely to Africa, eba‐140 seemed to be under stronger diversifying selection in South America than eba‐175. In contrast, eba‐181 did not show any sign of departure from neutrality. These changes in the patterns of selection on the eba genes could be the consequence of changes in the host immune response, the host receptor polymorphisms and/or the ability of the parasite to silence or express differentially its invasion proteins.     

Borrelia crocidurae in Ornithodoros ticks from northwestern Morocco: a range extension in relation to climatic change?

Tick‐borne relapsing fever (TBRF) is caused by Borrelia spirochetes transmitted to humans by Argasid soft ticks of the genus Ornithodoros. We investigated the presence of Ornithodoros ticks in rodent burrows in nine sites of the Gharb region of northwestern Morocco where we recently documented a high incidence of TBRF in humans. We assessed the Borrelia infection rate by nested PCR and sequencing. All sites investigated were colonized by ticks of the Ornithodoros marocanus complex and a high proportion of burrows (38.4%) were found to be infested. Borrelia infections were observed in 6.8% of the ticks tested. Two Borrelia species were identified by sequencing: B. hispanica and B. crocidurae. The discovery in northwestern Morocco of Ornithodoros ticks infected by B. crocidurae represents a 350 km range extension of this Sahelo‐Saharan spirochete in North Africa. The spread of B. crocidurae may be related to the increasing aridity of northwestern Morocco in relation to climate change.     

Cholesterol‐α‐glucosyltransferase gene is present in most Helicobacter species including gastric non‐Helicobacter pylori helicobacters obtained from Japanese patients

Background

Non‐Helicobacter pylori helicobacters (NHPHs) besides H. pylori infect human stomachs and cause chronic gastritis and mucosa‐associated lymphoid tissue lymphoma. Cholesteryl‐α‐glucosides have been identified as unique glycolipids present in H. pylori and some Helicobacter species. Cholesterol‐α‐glucosyltransferase (αCgT), a key enzyme for the biosynthesis of cholesteryl‐α‐glucosides, plays crucial roles in the pathogenicity of H. pylori. Therefore, it is important to examine αCgTs of NHPHs.

Materials and Methods

Six gastric NHPHs were isolated from Japanese patients and maintained in mouse stomachs. The αCgT genes were amplified by PCR and inverse PCR. We retrieved the αCgT genes of other Helicobacter species by BLAST searches in GenBank.

Results

αCgT genes were present in most Helicobacter species and in all Japanese isolates examined. However, we could find no candidate gene for αCgT in the whole genome of Helicobacter cinaedi and several enterohepatic species. Phylogenic analysis demonstrated that the αCgT genes of all Japanese isolates show high similarities to that of a zoonotic group of gastric NHPHs including Helicobacter suis, Helicobacter heilmannii, and Helicobacter ailurogastricus. Of 6 Japanese isolates, the αCgT genes of 4 isolates were identical to that of H. suis, and that of another 2 isolates were similar to that of H. heilmannii and H. ailurogastricus.

Conclusions

All gastric NHPHs examined showed presence of αCgT genes, indicating that αCgT may be beneficial for these helicobacters to infect human and possibly animal stomachs. Our study indicated that NHPHs could be classified into 2 groups, NHPHs with αCgT genes and NHPHs without αCgT genes.     

Value of heartburn for diagnosing gastroesophageal reflux disease in severely obese patients

Objective : To evaluate the prevalence of gastroesophageal reflux disease (GERD) in severely obese patients and the association between symptoms and objective data of GERD in this population. Research Methods and Procedures : A total of 158 consecutive severely obese patients (BMI ≥ 40 kg/m²) were prospectively evaluated. Symptoms were evaluated by a structured clinical questionnaire. Objective assessment was made by ambulatory 24‐hour esophageal pH monitoring and endoscopy. GERD was defined by the presence of symptoms or complications (esophagitis). The clinical criterion defining GERD was the presence of at least two episodes of heartburn per week. Results : The mean age of the 138 patients subjected to complete study was 42.6 ± 10.2 years, with a BMI of 50.1 ± 6.9 kg/m² (range, 40.6 to 69.4 kg/m²); 78% were women. The prevalence of GERD evaluated by symptoms and/or esophagitis was 33.3% (46/138). Clinical criteria of GERD were present in 31/138 cases (22.5%), and 26 (18.8%) had esophagitis. In 69/138 patients (50%), pHmetry was abnormal. Fifty‐three patients with esophagitis and/or abnormal pHmetry were asymptomatic. The sensitivity of heartburn as a diagnostic criterion of GERD in patients with severe obesity was 29.3%, with a specificity of 85.7%. No significant association was observed between severe obesity grade and the prevalence of symptoms and/or objective data. Discussion : Asymptomatic gastroesophageal reflux (abnormal esophageal acid exposure and/or reflux esophagitis) is more common than symptomatic gastroesophageal reflux in severely obese patients. Increased BMI is not associated with a greater prevalence of GERD in these patients.